000442   KIDOIKHAMMOUAN S, SEUBWAI W, SILSIRIVANIT A, WONGKHAM S, SAWANYAWISUTH K, WONGKHAM C (Forensic Medicine Dep, Khon Kaen Univ, Khon Kaen- 40002, Thailand, Email: wunchanas@yahoo.com) : Blocking of methionine aminopeptidase‑2 by TNP‑470 induces apoptosis and increases chemosensitivity of cholangiocarcinoma. J Can Res Ther 2019, 15(1), 148-52.

Resistance of cancer cells to chemotherapeutic drugs is a major pitfall of the failure of chemotherapy treatment for cholangiocarcinoma (CCA). A new therapeutic strategy that can improve treatment efficacy is mandatory for CCA patients. Our previous findings demonstrated the overexpression of methionine aminopeptidase‑2 (MetAP2) in CCA patients. In addition, supplementation of TNP‑470, a MetAP2 inhibitor, significantly inhibited the growth and metastatic activities of CCA cell lines. However, the molecular mechanism of antitumor activity of TNP‑470 and the synergistic antitumor activity of TNP‑470 combined with chemotherapeutic drugs are still unknown. The aim of this study is to evaluate the molecular mechanism of anticancer activity and the potential use of TNP‑470 as a chemosensitizing agent in CCA cell lines. Cell cycle and apoptosis of CCA cell lines were evaluated using flow cytometry with propidium iodide staining. Expression of apoptosis regulatory proteins was measured by Western blotting. The chemosensitizing effect of TNP‑470 was determined using combination index. TNP‑470 inhibited the growth of CCA cells via induction of apoptosis through activation of the p38‑phosphorylation and up‑ and down‑regulation of Bax and Bcl‑xL, respectively. Furthermore, TNP‑470 significantly enhanced the antitumor activity of 5‑fluorouracil, cisplatin, doxorubicin, and gemcitabine. The present results show that TNP‑470 could be a potential therapeutic or adjuvant agent for CCA.

2 illus, 2 tables, 27 ref

000449   MA Y, WANG B, LI L, WANG F, XIA X (Medicine Coll of Pingdingshan Univ, Pingdingshan 476000, Henan Province, China, Email: xiaxichao8336@163.com) : The administration of peroxisome proliferator‑activated receptors α/γ agonist TZD18 inhibits cell growth and induces apoptosis in human gastric cancer cell lines. J Can Res Ther 2019, 15(1), 120-5.

This study is to investigate the effects of a novel peroxisome proliferator‑activated receptor (PPAR) α/γ dual agonist TZD18 on cell growth, apoptosis, caspase activity, mitochondrial membrane potential, cytochrome c release, and apoptotic‑related protein expression in MKN‑45 cells. 3‑(4, 5‑dimethylthiazolyl)‑2,5‑diphenyltetrazolium bromide assay against various human cancer cell lines was performed to investigate the whether TZD18 could in reduce the proliferation rates of cancer cells. The percentages of apoptotic cells and mitochondrial membrane potential level were determined by flow cytometry. The subcellular localization of cytochrome c was examined by immunofluorescence microscopy. Western blotting assay was performed to reveal the expression of apoptosis‑related proteins. The results showed that the administration of TZD18 could inhibit the growth of MKN‑45 cells in a dose‑ and time‑dependent manner. In addition, the apoptotic ratio increased sharply along with a significant increase of caspase activities, mitochondrial membrane potential, and cytochrome c release following TZD18 exposure. The expression of Bax and p27kip1 increased significantly, whereas the expression level of Bcl‑2 protein was downregulated. These results indicated that the administration of PPAR α/γ agonist TZD18 may inhibit cell growth by inducing the apoptotic process in MKN‑45 cells.

5 illus, 30 ref

000392   ADAMI B, TABATABAEIAN H, GHAEDI K, TALEBI A, AZADEH M, DEHDASHTIAN E (Biology Dep, Isfahan Univ, Isfahan 81746-73441, Iran, Email: kamranghaedi@sci.ui.ac.ir) : miR-146a is deregulated in gastric cancer. J Can Res Ther 2019, 15(1), 108-14.

Gastric cancer is one of the most significant reasons for cancer‑related death. miR‑146a is one of the dysregulated factors associated with gastric tumorigenesis. However, deregulation of this microRNA (miRNA) has become controversial. Moreover, the inflammation‑mediating role of this miRNA implies that miR‑146a might be dysregulated by gastric cancer‑related pathogens, such as Helicobacter pylori. However, the dysregulation of miR‑146a in H. pylori‑infected gastric tumors has not been widely studied. We aimed to analyze the expression level of miR‑146a in gastric cancer tissues and then to assess any potential association between miR‑146a and H. pylori infection and other clinical characteristics. miR‑146a expression level was quantitatively studied by reverse transcription quantitative polymerase chain reaction, in 144 fresh tissues including 44 normal and 100 gastric cancer samples. A dramatic overexpression of miR‑146a was observed in primary gastric tumors. miR‑146a showed lower expression in progressed tumors with greater stages and lymph node metastasis. miR‑146a is highly expressed in primary gastric tumor independent of H. pylori infection. It is highly expressed in the lower stages and lymph node‑negative tumors. It might suggest the importance of upregulation and downregulation of this miRNA in the initiating/promoting and progressive steps of gastric tumorigenesis, respectively

4 illus, 2 tables, 37 ref

000430   HUANG J, WEI Y, ZHOU X, WANG L, HUANG M, WANG J (Colorectal Surgery Dep, The Sixth Affiliated Hospital of Sun Yat-Sen Univ, Guangzhou 510655, China, Email: wjp@mail.sysu.edu.cn) : The association between survivin −31G>C polymorphism and susceptibility to sporadic colorectal cancer in a Southern Chinese population. J Can Res Ther 2019, 15(1), 82-6.

The case–control study aimed to investigate the association between the −31G>C polymorphism in the promoter of survivin gene and the susceptibility to sporadic colorectal cancer (CRC) in a Southern Chinese population. The study was carried out on 711 healthy controls and 702 CRC cases of a Southern Chinese population. Survivin gene −31G>C genotypes were determined by polymerase chain reaction‑restriction fragment length polymorphism. The association between CRC risk and −31G>C genetic polymorphism was estimated using an unconditional logistic regression model. The number of CC genotype carried in CRC patients was much higher than those of controls (P < 0.001). Compared with CC genotypes, GC, GG genotypes and −31G wild‑type genotypes (i.e., GC + GG) had a significantly decreased risk of CRC (P < 0.001). In addition, survivin −31G wild‑type genotypes were not associated with decreased risk of sporadic CRC patients with body mass index (BMI) ≥28.0 kg/m², family cancer history, and premenopausal. Survivin −31G>C polymorphism is associated with sporadic CRC risk in the Southern Chinese population. The −31G wild‑type genotypes and GC, GG genotypes are the independent protective factors against sporadic CRC excluding those with a BMI ≥ 28.0 kg/m², family cancer history, and premenopausal.

3 tables, 31 ref

000406   CHAITRA L P, PRASHANT A, GOWTHAMI C S, HAJIRA B, SUMA M N, MAHESH S S, MANJUNATH G V, SHEELADEVI C S (Biochemistry Dep, JSS Medical Coll, Mysore - 570 015, Karnataka, Email: akila.prashant@gmail.com) : Detection of cancer stem cell‑related markers in different stages of colorectal carcinoma patients of Indian origin by immunohistochemistry. J Can Res Ther 2019, 15(1), 75-81.

Although the incidence rate of colorectal cancer (CRC) in all Indian cancer registries is very close to the lowest rate in the world, westernization has shown an increasing trend in the recent years. Recurrence is reported in CRC because the slowly proliferating stem cells escape the chemotherapeutic regimen. To detect the presence of CD133 and CD44 in human CRC specimens and to correlate the level of marker expression with tumor staging. We included 26 colorectal carcinoma patients between 20 and 70 years of age. Histological and immunohistochemical analysis of CD133 and CD44 was done in sections of 5 µm prepared from paraffin‑embedded blocks with most representative areas. All analyses were performed using Microsoft Excel 2010 and SPSS version 22. CD133 expression was seen exclusively on the cell membrane at the glandular luminal surface with dot‑like cytoplasmic staining. In the normal mucosa, CD44 expression was seen in the superficial region of the cell, whereas in most of the carcinomas, the staining was localized in the basolateral region of the cell. Both CD133 and CD44 showed significant correlation with tumor stage. In the present study, CD133 and CD44 show significant correlation with tumor staging. Cancer stem cell markers have shown similar pattern of expression in the patients of Indian origin. Using combination of markers for staging is preferred as it increases the sensitivity and specificity.

2 illus, 3 tables, 37 ref

000399   BAYRAM D, OZGOCMEN M, ARMAGAN I, SEVIMLI M, TUREL G Y, SENOL N (Histology and Embryology Dep, Süleyman Demirel Univ, Isparta-32260, Turkey, Email: dilekbayram@sdu.edu.tr) : Investigation of apoptotic effect of juglone on CCL‑228‑SW 480 colon cancer cell line. J Can Res Ther 2019, 15(1), 68-74.

Colon cancer is a major cause of morbidity and mortality in the world. Juglone is a natural compound which has been isolated from Juglans mandshurica Maxim, and it has various pharmacological effects such as antiviral, antibacterial, and anticancer. In our study, we aimed to investigate the effect of juglone on CCL‑228‑SW 480 colon carcinoma cell line in monolayer and spheroid culture medium. The CCL‑228‑SW 480 cell lines were cultured in both monolayer and spheroid cultures. Cells were treated with juglone at 24, 48, and 72 h of incubation. ID50 inhibition was determined on the dose for juglone and after it was found 20 μM was applied to the cells to examine the effect of juglone on CCL‑228‑SW 480 colon carcinoma cell line. After Juglone was applied the BrdU marking index, Transferase dUTP Nick ends Labeling (TUNEL) assay, active caspase‑3 assay, apoptosis‑inducing factor (AIF) assay were determined by immunohistochemistry in both the monolayer and spheroid cultures. The control group had a healthy pattern of S‑phase fraction, and many of the CCL‑228‑SW 480 cells nuclei were observed to be positive for BrdU. Terminal Deoxynucleotidyl TUNEL‑positive cells, active Caspase‑3, and AIF were detected after treatment with juglone in both the monolayer and spheroid cultures. The dead cell count was higher in the CCL‑228‑SW 480 cell lines with juglone applied than in the controls. Juglone significantly inhibits the proliferation and induces the apoptosis of CCL‑228‑SW 480 cells in vitro.

6 illus, 27 ref

000505   ZHAND S, HOSSEINI S M, TABARRAEI A , MORADI A, SAEIDI M (Microbiology Dep, Golestan Univ of Medical Sciences, Gorgan, Iran, Email: abmoradi@yahoo.com) : Analysis of poliovirus receptor, CD155 expression in different human colorectal cancer cell lines: Implications for poliovirus virotherapy. J Can Res Ther 2019, 15(1), 61-7.

Poliovirus (PV) receptor (CD155) is expressed on several kinds of cells and exerts diverse functions. Various investigations have confirmed that changes in CD155 expression in cancer cell lines affect metastasis, proliferation, and migration. The purpose of the present study was to investigate the CD155 transcript and protein expression in human colon adenocarcinoma cell lines in comparison to normal fetal human colon (FHC) cells. The CD155 expression level in four human adenocarcinoma cell lines and normal colon cell line were assessed using the SYBR green quantitative real‑time polymerase chain reaction (PCR) and flowcytometry. The results of real‑time PCR indicated that CD155 was significantly overexpressed in all human adenocarcinoma cell lines (P = 0.000). The highest and the lowest expression level of CD155 messenger RNA was observed in SW480 and HT29 cell lines by 491.14, and 12.04 fold changes, respectively, in comparison with the human normal cell line (FHC). Results of flowcytometry indicate that protein was strongly expressed in cancer cell lines. SW480 cells showed the highest CD155 protein expression level of 98.1%, whereas this protein expression was 1.3% in human normal colon cell line (FHC). Totally, these data indicate that CD155 expression is significantly elevated in cancer cell lines. The preferential expression of CD155 on cancer cell lines rather than on normal cell line suggests that CD155 could be targeted for future PV virotherapy.

3 illus, 3 tables, 37 ref

000424   HAN Z, HUANG H, ZHANG T (Medical Technology Dep, Zhengzhou Railway Vocational and Technical Coll, Henan- 450 052, P.R. China, Email: hanzhongmin1@163.com) : Downregulation of DBN1 is related to vincristine resistance in colon cancer cells. J Can Res Ther 2019, 15(1), 38-41.

This study was aimed to investigate the relationship between the expression of drebrin (DBN1) gene and resistance in colon cancer to reveal the mechanism of tumor drug resistance and provide a basis for the reversal of this drug resistance in tumor cells. The human colon carcinoma cell line HCT‑8 was used, and vincristine (VCR)‑resistant colon cancer cell line HCT‑8/V was established by gradually increasing the concentration of VCR. Polymerase chain reaction (PCR) primers were designed for DBN1 gene. The DBN1 differential expression in colon cancer sensitive and resistant cell lines was detected by fluorescence quantitative PCR. Western blot analysis was used to study DBN1 expression in the resistant cells further. VCR resistance of HCT‑8/V cell line was established. Quantitative PCR and Western blot results showed that DBN1 expression in the resistant cell line was significantly lower, the difference being statistically significant (P < 0.05). Low DBN1 gene expression may be associated with colon cancer cell resistance to VCR.

2 illus, 24 ref

000489   SOLTANI-SEDEH H, IRANI S, MIRFAKHRAIE R, SOLEIMANI M (Biology Dep, Islamic Azad Univ, Tehran, Iran, Email: s.irani@srbiau.ac.ir) : Potential using of microRNA‑34A in combination with paclitaxel in colorectal cancer cells. J Can Res Ther 2019, 15(1), 32-7.

MicroRNAs are small noncoding RNAs which modulate gene expression at different levels. It has been shown that downregulation of miR‑34a occurs in varieties of cancers including colorectal cancer (CRC). In this study, we investigated the potential tumor inhibitory effects of miR‑34a alone or in combination with paclitaxel in CRC cells. SW480 cells were transduced with lentiviral overexpressed miR‑34a. First, using 3‑(4,5‑Dimethyl‑2‑thiazolyl)‑2,5‑diphenyl‑2H‑tetrazolium bromide assay, the effect of miR‑34a induction alone or in combination with paclitaxel on the cell viability and cell proliferation were estimated. Then, the expression level of target genes was measured using quantitative reverse transcription‑polymerase chain reaction analysis. Eventually, the role of miR‑34a and paclitaxel on cell cycle were determined with flow cytometry. Gene expression analysis showed that miR‑34a downregulates the expression of BCL2 and SIRT1 genes at mRNA level. Furthermore, miR‑34a has a potential to reduce cell viability and cell cycle arrest at G1 phase. Combination of paclitaxel with overexpression of miR‑34a significantly decreased cell viability compared to cell treated with miR‑34a or paclitaxel alone. Interestingly, a combination of miR‑34a and paclitaxel arrested cell cycle at two phases. Our results suggested that combination therapy of miR‑34a and paclitaxel could be considered as the potential treatment of CRC.

5 illus, 1 table, 28 ref

000398   BAHADIR A, ERAL G, BUDAK M, SHIMAMOTO F, KORPINAR M A, ERDAMAR S, TUNCEL H (Biophysics Dep, Istanbul Univ, Istanbul- 34303, Turkey, Email: hntuncel@istanbul.edu.tr) : Association of clinicopathological features with E‑cadherin (CDH1) gene‑160 C>A promoter polymorphism in Turkish colorectal cancer patients. J Can Res Ther 2019, 15(1), 26-31.

The role of E‑cadherin (CDH1) gene‑160 C>A (rs16260) promoter polymorphism in colorectal cancer (CRC) still remains inconclusive. The aim of this study is to investigate the associations between the CDH1‑160 C>A polymorphism with the susceptibility and clinicopathological development of CRC in the Turkish patients. To our knowledge, this is the first report examining the role of CDH1 polymorphism in Turkish CRC patients. A total of 92 colorectal carcinoma cases (including 62 colon and 30 rectal cancer patients) and the corresponding adjacent normal tissues as controls were studied. The polymorphism was genotyped using polymerase chain reaction‑restriction fragment length polymorphism analysis. Clinicopathological features including patient’s age, gender, tumor stage, and tumor location (colon/rectum) were compared statistically with the polymorphism status. There was no significant difference in both genotype and allele frequencies of the CDH1 polymorphism between colorectal tumor cases and normal samples (P = 0.472 and 0.508, respectively). Furthermore, no significant associations were observed between the CDH1 polymorphism status and age, gender, tumor stage, and tumor location of the colorectal tumor cases(all P > 0.05). These results indicate that CDH1‑160 C>A polymorphism does not contribute to the genetic susceptibility of CRC and the polymorphism may not be a direct effect on the progression of the disease in Turkish CRC patients.

4 tables, 33 ref

000446   LENZI P, STOEPLER T M, MCHENRY D J, DAVIS R E, WOLF T K (Biology Dep, Radford Univ, Radford, VA 24142, Email: plenzi@radford.edu) : Jikradia olitoria ([Hemiptera]:[Cicadellidae]) transmits the sequevar NAGYIIIβ phytoplasma strain associated with North American grapevine yellows in artificial feeding assays. J Insect Sci 2019, 19(1), 1.

North American Grapevine Yellows (NAGY) is a destructive disease of grapevines caused by phytoplasmas, wallless bacteria that are insect-transmitted and found in plant phloem tissues. Although the disease was recognized in vineyards in the eastern United States since the 1980s, the identities of vectors remain unknown. The objectives of this study were to survey potential phytoplasma vector insects inhabiting Virginia vineyards that expressed NAGY symptoms and to evaluate their ability to transmit phytoplasmas associated with NAGY. Phytoplasmas were identified as ‘Candidatus Phytoplasma pruni’-related NAGYIIIβ strains and ‘Ca. Phytoplasma asteris’-related NAGYI-B strains. To determine the identities of the potential vectors, artificial feeding solution was used to evaluate the ability of leafhopper species to release phytoplasmas during feeding and phytoplasma strains were identified using molecular tools. Out of 49 insect species screened, Jikradia olitoria was the only insect that released phytoplasmas into the feeding solutions; all phytoplasmas, thus, detected were identified as NAGYIIIβ strains by nucleotide sequencing of three different genomic regions. No NAGYI-B strain was detected. To our knowledge, this is the first evidence of a potential insect vector of a specific phytoplasma associated with NAGY disease, and it is the first report of J. olitoria being a putative vector of a plant pathogenic phytoplasma.

1 illus, 4 tables, 34 ref

000448   LOKHANDE R V, AMBEKAR J G, BHAT K G, DONGRE N N (Biochemistry Dep, Maratha Mandal’s NGH Institute of Dental, Sciences and Research Centre, Belagavi, Karnataka, Email: ranivl@rediffmail.com) : Interleukin‑21 and its association with chronic periodontitis. J Indian Soc Periodontol 2019, 23(1), 21-4.

Interleukin‑21 (IL‑21) is a pleiotropic cytokine, well documented to contribute to the development of Th17 cells which have been shown to play an important role in the pathogenesis of periodontitis. Periodontal disease is a chronic infection of tooth‑supporting tissue. This study evaluates the saliva and serum levels of IL‑21 in patients with chronic periodontitis and periodontally healthy individuals. The present study was carried out in the Department of Microbiology in association with Department of Oral Medicine and Radiology, Maratha Mandal’s N.G.H Institute of Dental Sciences and Research Centre, Belgavi, Karnataka. Fifty samples of each group were included in the present study. The levels of IL‑21 were assessed using a commercially available enzyme‑linked immunosorbent assay (ELISA) kit and the results were expressed as pg/ mL. Statistical analysis was performed using SPSS 17.0 software. Data were expressed as mean ± standard deviation and interquartile ranges and comparison of controls and cases by Mann–Whitney test. Serum and salivary levels of IL‑21 were significantly higher in chronic periodontitis group than in controls (P < 0.001). Clinical periodontal parameters correlated positively with serum IL‑21 levels. IL‑21 is highly expressed in patients with chronic periodontitis and correlated well with clinical parameters of periodontal destruction. Therefore, IL‑21 appears to play a role in tissue destruction and can be used as diagnostic biomarker in chronic periodontitis. Saliva can be considered to be a useful alternative to serum as a diagnostic sample.

4 tables, 22 ref

000483   SHEFALI, YADAV J, GUPTA R K (Zoology and Aquaculture Dep, CCS Haryana Agricultural Univ, Hisar, Haryana, Email: shefaligulliya@gmail.com) : Assessment of chromium induced alterations on gut bacterial population of E. eugeniae. J Appl Nat Sci 2019, 11(1), 94-6.

The aim of the present study was to enumerate the diversity of gut bacterial population of epigeic earthworm species, E. eugeniae exposed to heavy metal. Adult earthworms were treated with three different doses (0.06 %, 0.13 % and 0.19 % w/v) of potassium dichromate (K₂Cr₂O₇) for this purpose. The gut samples were serially diluted and inoculated in seven distinct media viz. Jensen's media, King’smedium B base, Burk’s media, Yeast Mannitol Agar media, Vogel- Johnson agar base media, Luria Bertani Broth Miller and Nutrient agar media. Dose dependent decrease in bacterial population was observed in treated worms when compared with control. Maximum percent change (62.59 %) in bacterial population was observed in Burk’s media when exposed to 0.19 % chromium. The results revealed that chromium alters the microbial population present in gut of earthworms. Therefore, changes in earthworms’ gut microbial community due to soil management practices can also be used as markers of soil fertility and quality.

2 illus, 1 table, 21 ref

000412   DEVI S U, MUTHUCHELIAN K (Microbiology and Biochemistry Dep, Nadar Saraswathi Coll of Arts and Science, Theni - 625 531, Email: spu.uma@gmail.com) : Analysis of physico-chemical parameters of soil and population density of Azospirillum isolates from paddy fields of Theni district, Tamilnadu. J Appl Nat Sci 2019, 11(1), 88 - 93.

The rhizosphere region of eight different paddy field areas of Theni district was studied for their physico-chemical analysis of soil and the population density of Azospirillum sp. The study results showed range of values of pH (7.4-7.9), bulk density (1.13g/cm³ -1.60g/cm³), water holding capacity (34.31 % - 18.25 %), electrical conductivity (1.31 – 1.11), organic carbon (0.93 % - 0.71 %) . The macronutrient values namely total nitrogen (1.72 % - 0.78 %), phosphorus content (0.177 % - 0.122 %) and potassium (1.364 % - 1.273 %) were observed. Also micronutrients of various paddy fields like Zn (0.9 % - 0.5 %), Cu (2.3 % - 1.7 %), Fe (10.9 % -8.2 %), Mn (6.7 % - 5.2 %) were recorded, whereas the values of available macronutrients namely nitrogen (295 kg/ha - 223 kg/ha), phosphorous (89 Kg/acre - 49 kg/acre), potassium (790 kg/acre - 490 kg/acre) were noted. The Azospirillum population density was highest in Chinnamanur (192 x10⁵ CFU/g) and lowest in Royyanpatti (91 x10⁵ CFU/g). Thus through this work we were able to isolate and identify novel high yielding Azosprillium sp from paddy fields of Theni district.

2 illus, 4 tables, 29 ref

000454   MAQBUL M S, IKBAL A R, MOHAMMED T, DAWOUD A, KHAN A A, IQUBAL S M S, KHAN K A, MUDDAPUR U M, SHEIK G B, SINGH S K (General Science Dep, Ibn Sina National Coll of Medical Sciences, Jeddah 21418, Kingdom of Saudi Arabia, Email: shakeeliqubal@gmail.com) : Determination of biochemical constituents of Argan essential oil and its antimicrobial efficacy against tinea infections of Trichophyton rubrum. Int J Green Pharm 2019, 14(3), 286-92.

The aim of this study to focus on these tinea infections caused by the mold Trichophyton rubrum by collecting the clinical specimens by means of skin scraping samples from the affected sites of the patients and to evaluate the susceptibility of the isolates of the obtained clinical specimens toward the Argan essential oil. A rapid phytochemical analysis study was also performed. Argan essential oil procured from Jeddah local market, clinical skin scrape samples from the patient. The phytochemical study was done to find out the chemical compounds present in the Argan essential oil, which plays a key role in determining the antimicrobial efficacy of the Argan essential oil. The interpretation of the observation and results for the Argan essential oil showed the promising study results. Regarding its efficacy as potential antifungal agents when compared to that of the standard synthetic chemical agents used against the clinical skin scrape isolates of T. rubrum. The phytochemical compounds present in the Argan essential oil acts as an effective remedy toward the clinical skin scrape isolates of T. rubrum compared to the standard antifungal agents.

1 illus, 4 tables, 20 ref

000432   HUGUETTE T T, TOM N L E, FLORENCE N T, FAROUCK A B, JOSEPH N, THÉOPHILE D (Biological Sciences Dep, Yaounde I Univ, Yaoundé, Cameroon, Email: dimo59@yahoo.com) : Preventive effects of aqueous extract of the whole plant of Eleusine indica (Linn) Gaertn. (Poaceae) against L-NAME induced nephrotoxicity in rat. J Phytopharmacol 2019, 8(1), 28-32.

The entire plant of Eleusine indica is used in Cameroonian folk medicine to treat several diseases including renal problems. We investigated the preventive effects of Eleusine indica aqueous extract (EIAE) against L-NAME induced renal damages in rats. Animals were divided into a control group (0.9 % saline, 10 mL/kg, ip) and 4 experimental groups. The rats in the negative control group received L-NAME (30 mg/kg, ip). The positive group was treated with losartan (12.5 mg/kg, per os) and L-NAME while the two last groups received EIAE (100 mg/kg or 200 mg/kg, per os) and L-NAME injection. After 60 days of treatment, blood pressure was measured; lipid profile, kidney and some oxidative stress parameters were evaluated. Intraperitoneal injection of L-NAME induced a significant elevation in blood pressure, of total cholesterol, triglycerides and LDL-c, with a significant reduction of HDL-c levels as compared to the control groups. Nephrotoxicity was evidenced by significant elevation in serum levels of creatinine, urea and K⁺ , accompanied by a significant reduction of Na⁺ and GFR as compared to controls. Catalase, GSH and nitrites were significantly decreased in L-NAME injected group. L-NAME solution gave simultaneously with Eleusine indica prevented the rise of blood pressure, improve lipid profile, kidney function and antioxidant defenses. These results confirm the nephroprotective effects of Eleusine indica aqueous extract and highlight its protective properties in L-NAME-induced renal failure and its antioxidant capacities against kidney damages.

2 illus, 2 tables, 35 ref

000457   MATAWALI A, LEE P C, HOW S E, JUALANG A G (Malaysia Sabah Univ, Sabah, Malaysia, Email: azlanajg@ums.edu.my) : Biological activities of Chromolena odorata (L.) King and Robinson (Asteraceae) collected from Sabah, Malaysia as protein phosphatase type-1 inhibitor. J Phytopharmacol 2019, 8(1), 01-4.

Chromolena Odorata has been traditionally used as wound healer in local community. The present study investigated the anti-kinase and anti-phosphatases activities on methanolic C. odorata extract. Mutant yeast strains used are MKK1^P386, MKK1^P386_MSG5, PAY704-1 and PAY700-4. Bioassay guided fractionation of C. odorata revealed positive activities of hexane, ethyl acetate and chloroform partitions. Column chromatography of all partitionates later confirmed fraction F2 from chloroform extract had most favorable activity with inhibitory zone ranged between 7 ± 0.0 mm until 15 ± 0.0 mm. Kinetic analysis including maximum enzyme velocity (V_m) and Michealis-Menten constant (K_m) were evaluated and compared for both normal and inhibited reactions. Enzyme activity with DiFMUP as substrate showed fraction F2 act as PP1 enzyme inhibitor with the K_m value 0.60 mM and V_m value 200 mM/min as compared to the normal enzymatic reaction. Results provided unveil the potential of C. odorata as an effective therapeutic agent.

2 tables, 33 ref

000390   ABDULLAHI A I, MODU M B, HARUNA M Y (Biology Dep, Ahmadu Bello Univ, Zaria, Nigeria) : Effect of bitter leaf (Vernonia amygdalina) extract on microbial stability of smoked-dried African catfish Clarias gariepinus (Burchell, 1822). Int J fish Aquat Stu 2019, 7(1), 299-303.

The paper evaluates the stability of microbes on of smoked-dried African catfish (Clarias gariepinus) treated with different concentrations of bitter leaf (Vernonia amygdalina) extract. The experimental treatments were the control, 0.5 %, 1 %, 1.5 % (w/v) bitter leaf extract solutions. A total of fifty two fish of average mean weight of 30.98 ± 1.32 g were gutted, washed and randomly assigned to the treatments. Thereafter the fish were soaked in the treatments for 30 minutes and later smoked-dried for 12 hours. After smoking the fish were stored in cartons and placed on laboratory table for 21 days. Six bacterial species namely, Bacillus subtilis, Corynebacteria sp, Proteus mirabilis, Streptococcus faecalis, Staphylococcus albus and Staphylococcus aureus were observed in the study. There was no increase in the microbial loads after 21 days of storage in samples treated with 0.5 % 1.0 % and 1.5 % solutions of the bitter leaf extract, but there was significant increase in control from 27×10³ cfu to 40×10³ cfu. The sample treated with 1.5 % solution of the bitter leaf extract exhibited the highest antibacterial effect and gave the best result. Therefore, V. amygdalina extract solution could be used in protecting stored smoked-dried catfish from bacterial spoilage, thus limiting economic loss and possible health risk to consumers.

1 illus, 4 tables, 17 ref

000461   MUKWABI D M, OKEMO P O, OTIENO S O, ODUOR R O, MACHARIA C M (Kenya Fisheries Service, Kenya) : Temperature and pH influence on bacterial pathogens infecting farmed Nile tilapia in aquaculture systems in Bungoma County, Kenya. Int J Fish Aquat Stud 2019, 7(1), 190-7.

Aquaculture production has been expanding in recent years through Governement support via financial and input subsidies in small scale farmers. However, fish farming just like other productive sectors face challenges which include fish diseases that constitute the largest cause of economic loss in aquaculture. The fish bacterial infections are caused by Aeromonas, Edwardsiella, Flavobacterium, Francisella, Photobacterium, Piscirickettsia, Pseudomonas, Tenacibaculum, Vibrio, Yersinia, Lactococcus, Renibacterium, Streptococcus, Nocardia, Staphylococcus, Vagococcus and Weisella species. This study focused on characterizing pathogenic bacteria present in aquaculture systems in Bungoma County. The sampled fish were transferred to a microbiology laboratory in Nairobi where they were subjected to bacterial analysis including microscopy, culture methods and biochemical tests. The study showed that temperature and pH were outside the recommended levels of fish culture production. Further, pathogenic bacteria recovered were Vibrio vulnificus, Vibrio parahaemolyticus, Aeromonas hydrophila, and Pseudomonas aeruginosa from Nile tilapia. Vibrio vulnificus and Vibrio parahaemolyticus were isolated from gills and were found to be significantly different (P<0.001) across the six sub counties. In fish ponds water, Aeromonas hydrophila were isolated and found to be significantly different (P<0.016) across the sub counties. In addition, Streptococcus iniae were isolated from fish feeds. Therefore, efforts should be made to reduce the presence of bacterial pathogens in aquaculture in order to increase fish yields.

5 tables, 47 ref

000495   TALUKDER B, CHAKMA F, SABUJ A A M, HAQUE Z F, RAHMAN M T, SAHA S (Microbiology and Hygiene Dep, Bangladesh Agricultural Univ, Mymensingh-2202, Bangladesh) : Identification, prevalence and antimicrobial susceptibility of major pathogenic bacteria in exportable shrimp (Penaeus monodon) of southern Bangladesh. Int J Fish Aquat Stud 2019, 7(1), 96-9.

A total of 30 samples of shrimp were collected from two local sea food processing plant of Rupsha and Mongla Bazar of Khulna district to investigate the microbiological quality. Microbial quality was assessed by total viable count (TVC), total Vibrio count (TViC), total Staphylococcal count (TSC) and total E. coli count (TEC) by inoculating into specific media and the values of all the samples were ranged from log 2.92 to 3.41 cfu/gm, log 2.06 to 2.11 cfu/gm, log 2.21 to 2.49 cfu/gm and log 2.14 to 2.22 cfu/gm respectively. Out of 30 samples 10, 11 and 5 samples were found positive for Vibrio spp., Staphylococcus spp. and. E. coli respectively. Results of antibiotic susceptibility test against eight commercially available antibiotics showed all three bacteria were multidrug resistant. This study revealed that the shrimp samples of Mongla contain higher load of bacteria than the samples from Rupsha.

4 illus, 2 tables, 24 ref

000479   SASIKALA S, NAIDU M D (Biotechnology Dep, Sri Venkateswara Univ, Tirupati, Andhra Pradesh) : Evaluation of protective effect of Centella asiatica leaves on pancreas function in diabetic rats. Int J Herb Med 2019, 7(1), 55-60.

Centella asiatica is a well-known plant with a wide range of medicinal properties. The aim of the study is to investigate the protective effect of Centella asiatica leaves on pancreas function in experimental diabeticrats. Diabetes was induced in rats by injecting streptozotocin at a dose of 50 mg/kg body weight (b.w) intraperitoneally. The experimental rats were treated with the methanol extract of Centella asiatica leaves (CALEt) at a dose of 300 mg/kg b. w. After the treatment period all the animals were sacrificed, and the blood sample was analyzed for insulin, glucagon, glycosylated haemoglobin and other blood parameters. Oral administration of CALEt for a period of 30 days restored the altered parameters. Histology of diabetic rats treated with CALEt showed pancreatic β-cell regeneration. Thus, our results revealed that the administration of CALEt may have protective effect on the pancreas of STZ induced diabetic rats.

7 illus, 2 tables, 14 ref

000464   N’DILLE G M, BIYITI L F, FROMME T, OYONO J L E, KLINGENSPOR M (Technical Univ of Munich, GregorMendel-Str. 2, Germany) : Ethanolic extract of Iwong (Ipomoea alba L., Convolvulaceae) attenuates adipogenesis in 3T3-L1 adipocytes. Int J Herb Med 2019, 7(1), 11-7.

Several traditional medicinal plants have been investigated with respect to their anti-obesity potential. Ipomoea alba (common names: Iwong or Moon Wine) is used in Cameroonian traditional medicine as a laxative and anti-diabetic agent as well as to promote weight loss and improve breast milk quality. The goal of our study was to evaluate the effect of Iwong in cultured adipocytes. We assessed the effect of ethanolic extracts of Iwong in vitro on adipogenesis in the 3T3-L1 preadipocyte cell line. Treatment of 3T3-L1 cells with ethanolic extracts of Iwong significantly reduced lipid accumulation after 9 days. This reduction was associated with a significant decrease in glycerol-3-phosphate dehydrogenase (GPDH) activity and PPARγ and C/EBPα mRNA levels. The anti-adipogenic effect of Iwong was restricted to the early phases of adipogenesis. Our results indicate that Iwong contains natural substances potentially suitable for the treatment of obesity via inhibition of lipid accumulation.

5 illus, 1 table, 25 ref

000458   MISHRA O P, CHHABRA P, NARAYAN G, SRIVASTAVA P, PRASAD R, SINGH A, ABHINAY A, BATRA V V (Pediatrics Dep, Banaras Hindu Univ, Varanasi, Email: opmpedia@yahoo.co.uk) : Cytotoxic T- lymphocyte antigen-4 (CTLA4) gene expression and urinary CTLA4 levels in idiopathic nephrotic syndrome. Indian J Pediatr 2019, 86(1), 26–31.

To detect Cytotoxic T- Lymphocyte Antigen-4 (CTLA4) single nucleotide polymorphisms (SNPs) at +49A/G (rs231775) and -318C/T (rs5742909) positions in children with idiopathic nephrotic syndrome (INS) and also assay urinary soluble CTLA4 (sCTLA4) levels in children with minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS) and steroid sensitive nephrotic syndrome (SSNS) in remission. The study included 59 patients of INS (MCD-23, FSGS-15 and SSNS in remission-21) and 35 healthy controls. The CTLA4 SNPs profiling was done in peripheral blood mononuclear cells and urinary sCTLA4 level was assayed by ELISA kit. Although frequency of homozygous +49 GG (rs4553808) genotype (26.3 % vs. 11.4 %; p = 0.231) and G allele (52.6 % vs. 40 %; p = 0.216) were found to be higher in INS as compared to controls, the differences were statistically non-significant. Genotypes GG, AG, AA and alleles A and G frequencies were comparable among MCD, FSGS and controls. SNP at −318 C/T (rs5742909) did not show homozygous TT genotype both in INS as well as controls. Median urinary sCTLA4/creatinine level was significantly higher in MCD as compared to FSGS (p = 0.027), SSNS in remission (p = 0.001) and controls (p = 0.003). The positive associations of +49 GG genotype and G allele in patients with nephrotic syndrome were not observed. The frequencies did not differ significantly among MCD, FSGS and controls. Urinary sCTLA4 level was significantly increased in MCD; suggesting its possible role in the pathogenesis of disease.

3 illus, 2 tables, 24 ref

000441   KHORASANI M M Y, YOUSEFI A, ZAINODINI N (Endodontics Dep, Rafsanjan Univ of Medical Sciences, Rafsanjan, Iran, Email: m.yaghooti@yahoo.com) : NLRP1 and NLRC4 inflammasomes are not responsible for the induction of inflammation in pulp tissues from carious teeth. J Conserv Dent 2019, 22(1), 12-6.

Inflammation is a risk factor for dental complications. Inflammasomes are a set of intracellular sensors which participate in the induction of inflammation. As the main factors involved in the induction of pulp inflammation in the carious teeth are yet to be clarified, this study was aimed to evaluate NLRP1 and NLRC4 expression levels in the inflamed and healthy pulps. Fifty inflamed and fifty healthy pulps were evaluated regarding the multiRNA levels of NLRP1 and NLRC4 using real‑time polymerase chain reaction technique. Results demonstrated that expression of NLRP1 (P = 0.985) and NLRC4 (P = 0.581) did not significantly differ in inflamed in comparison to healthy pulps. NLRP1 (P = 0.989) and NLRC4 (P = 0.170) did not change in males when compared with females in inflamed pulps. Furthermore, NLRP1 (P = 0.133) and NLRC4 (P = 0.642) were not altered in males in comparison to females in healthy pulps. Although NLRP1 and NLRC4 are the main inflammasomes, it appears that they are not the responsible molecules involved in the human pulp inflammation in the carious teeth.

3 tables, 24 ref

000416   FILIPTSOVA O, LITOVCHENKO Y, NABOKA O, LUCHKO E, DYOMINA Y, GALIY L, BUDANOVA L, FILYANINA N (Biology Dep, National Univ of Pharmacy, Kharkov, Ukraine, Email: philiptsova@yahoo.com) : Facial asymmetry in Slavic populations: Sex dimorphism in healthy young Ukrainians. J Anat Soc India 2019, 68(1), 68-73.

Facial and body asymmetry to some extent is present in all individuals. Because fluctuating asymmetry is of a stochastic nature, it cannot be studied at the level of individuals, but only its groups can be considering during studying the populations. We conducted a survey of 200 students of 8–11 grades, from 13 to 17 years old, residents of Ukraine. Asymmetry measurements were made in the program GIMP‑2.8. Population distribution of eight points of the face asymmetry among the population of Ukraine was studied. Sex dimorphism was found under all average indicators of absolute and relative asymmetry. Higher indicators of asymmetry were common to males and also typical for the lower part of the face. Obtained data complement the existing study on facial asymmetry in human. Some of our results are consistent with the existing data from previous studies of the world population, while others point to the differences of Slavic population in a number of features related to the facial asymmetry.

1 illus, 2 tables, 47 ref

000503   YILMAZ S, ALPA S, NISARI M, KARATOPRAK G S, DOgANYIgIT Z, ÜLGER H, ERTEKIN T (Anatomy Dep, Bozok Univ, Yozgat, Turkey, Email: sehery38@hotmail.com) : Examining the antitumoral effect of Cornelian cherry (Cornus mas) in ehrlich ascites tumor induced mice. J Anat Soc India 2019, 68(1), 16-22.

Different doses of C. Mas concentrated syrup on ascitic tumors was investigated in the Ehrlich Ascites Tumor model (EAT). A total of 46 Balb/C mice were used in our study, 6 of which were stock animals and the other were in ascitic tumor groups. EAT cells (1x106 EAT cells) were injected intraperitoneally into all of the mice. Mice in the treatment groups with ascitic tumors received 100 mg/kg and 200 mg/kg Cornus Mas extract intraperitoneally for 9 days. Counts after the 3 and 24-hour incubations in the EAT cell line that the average number of the dead cells was less in the group to which 100 µg/ml C. Mas was administered when compared with the control group, and that this difference was significant at a statistical level (P < 0.05). The purpose was also to determine the in vitro cytotoxic effects of Cornus Mas on EAT cells, to define the alive and dead cell rates, and to compare the 3-hour and 24-hour incubation in groups to which Cornus Mas (syrup) extract were given at different concentrations (50, 100, 250 μg/ml). EAT model is one of the animal tumors induced empirically, it has been the subject matter of many other studies. In the group in which EAT was applied together with high-dose C. mas fruit syrup, it was observed that the EAT cells were not as intense as they were in the tumor control group. Our study showed the anti-tumor effect of C. Mas in assisted tumor development with EAT cells.

5 illus, 5 tables, 24 ref

000433   JEELANI K, VASUDEVA N, MISHRA S (Anatomy Dep, Maulana Azad Medical Coll, New Delhi-110 002, Email: sabitamishra12@gmail. com) : Expression of fibroblast growth factor 10 during histogenesis of human lung in prenatal period. J Anat Soc India 2019, 68(1), 7-11.

The developing human lung has five stages. Many important signaling molecules have been identified at each stage of the developing lung. Fibroblast growth factor (FGF) signaling, mainly FGF10, is a key molecule in airway branching. Failure of the expression of FGF10 may lead to hypoplastic lung and blunted tracheal end. In this view, the expression of FGF10 was studied in each stage of human lung development. In the present study, ten aborted fetuses of gestational age 14–26 weeks were procured from the Department of Obstetrics and Gynaecology, LNH, New Delhi, after ethical clearance. After fixation in 10 % formalin, serial sections of each lung tissue were stained with hematoxylin and eosin, and immunostaining was done to see the expression of FGF10. The amount of mesenchymal tissue decreased and condensed with advancing gestational age. At 14‑week gestation, the first expression of FGF10 was seen in mesenchyme by immunostaining. As the gestational age advances, the amount of mesenchymal tissue decreases and gradual diminution of the height of epithelium was seen. By the 26th week of intrauterine life, fetal lung attains morphological maturity so as to support the extrauterine life of a growing fetus.

3 illus, 1 table, 21 ref

000429   HEYDARIZADI S, ABBASI N, ASADOLLAHI K, REZAEE S, MORADIPOUR A , AZIZI M (Anatomy Dep, Ilam Univ of Medical Sciences, Ilam, Email: azizi.moaz@gmail.com) : Effects of transplanted olfactory ensheathing cells on functional improvement and axonal regeneration in acute and delayed spinal cord injury in rats: A comparative study. J Anat Soc India 2019, 68(1), 1-6.

Cell therapy is known as one of the most common methods used for treating a spinal cord injury (SCI); particularly, olfactory ensheathing cells (OECs), which have attracted much more attention among scholars due to their properties such as promotion of axonal regeneration, remyelination, and angiogenesis. Thus, the present study compared the effects of transplanted OECs on functional improvement and axonal regeneration of contused rats during acute and delayed phases. For this purpose, a total of 56 male Wistar rats were randomly divided into eight groups including sham, control, three vehicle groups (immediately, 3 and 7 days after injury), and three cell transplantation groups (immediately, 3 and 7 days following injury). The sham group had experienced only laminectomy and other groups had undergone the SCI. The olfactory mucosa of the 7‑day‑old rat pups was also used for cell culture. The cell type was then confirmed by immunocytochemistry. In the vehicle and cell transplantation groups, the cell culture medium was injected by itself or accompanied by cells, respectively. Subsequently, the motor function was evaluated. Finally, luxol fast blue (LFB) staining was used for histological assessment. Motor test results showed an increase in the Basso, Beattie, and Bresnahan locomotor scale scores immediately after injury in the transplantation group compared to those in other two cell‑treated groups; but it was not significant. Based on the LFB staining results, the regeneration rate in the transplantation group immediately after injury was considerably higher than that in two other treated groups. Therefore, considering these findings, it seemed that cell transplantation immediately after injury was better than that 3 and 7 days following injury.

4 illus, 27 ref

000459   MOHAMMED N M H , EL-DRIENY E , EL-DRUSSI I S , AL-AGORY M, GHETH E M M (Histology Dep, Omar Al-Mukhtar Univ, El-Beida, Libya) : Histopathological changes in liver tissue induced by meloxicam in male mice. Int J Pharm Life Sci 2019, 10(1), 6059-63.

Despite of the frequent utility of non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of osteoarthritis related diseases, as they have analgesic antipyretic and anti-inflammatory effects, they have been currently proven to cause deleterious toxic effects to the hepatic tissue. Meloxicam is a new version of NSAIDs classified as selective cyclooxygenase-2 inhibitor (COX-2) which may have a protective role over the old traditional types (non-selective COX-1 inhibitor), therefore, the aim of the study was to investigate whether meloxicam could have toxic pathological effects on the liver tissue like other traditional drugs. Fifteen adult male mice were divided into two groups, one group of five mice served as control and received only distilled water, while the animals of the second group (10 mice) was treated with meloxicam 0.4 mg/kg administrated daily by oral gastric gavage for 10 days. After 24 hours of the last dose, animals were sacrificed, their livers were removed and processed for histological examination by light microscope. The liver sections obtained from meloxicam treated mice showed dilatation and congestion of blood sinusoids, as well as portal venous congestion. Appearance of degenerative changes of hepatocytes was found in forms of cytoplasmic vacuolations and nuclear pyknosis as well as glycogen depletion. In addition, mononuclear cellular infiltration, areas of hemorrhage and necrosis were evident. Focal loss of normal hepatic architecture was also observed. It could be conclude from this study that meloxicam might cause mild to moderate hepatotoxicity.

4 illus, 17 ref

000452   MANDLOI P, PANDEY R (Pharmacology Dep, Swami Vivekanand Coll of Pharmacy, Indore, Madhya Pradesh, Email: pmandloi025@gmail.com) : Evaluation of anti-anxiety activity of Lawsonia inermis Linn. Int J Pharm Life Sci 2019, 10(1), 6016-9.

Anxiety is one of the most common mental disorders, characterized by changes in mood, behavior, somatic function, and cognition. Benzodiazepines and SSRIs are most commonly employed drugs for the treatment of anxiety. Synthetic drugs available for treatment of anxiety have various adverse effects. Drugs obtained from natural sources are known to cause fewer side effects compared to synthetic drugs despite of same ability to cure disease. Lawsonia inermis commonly known as henna is a perennial herbaceous plant belonging to family Lythraceae. Traditionally it has been used to treat skin diseases, dysentery, bronchitis, anemia and inflammation. The aim of present study was to investigate the anxiolytic activity of methanolic extracts of Lawsonia inermis leaves in mice. Methanolic extracts were administered orally at a dose of 200 and 400 mg/kg bw. The results were analyzed by One Way Analysis of Variance (ANOVA).The results showed the extracts of leaves showed significant anxiety relatively methanolic extract. The findings concluded that Lawsonia inermis leaves exhibit anxiety and further studies are suggested to isolate the active principles responsible for the activity.

3 tables, 5 ref

000408   CHAWDA N, PANDEY R (Pharmacology Dep, Swami Vivekanand Coll of Pharmacy, Indore, Madhya Pradesh, Email: nidhichawda3031@gmail.com) : Evaluation of analgesic activity of fruits of Semecarpus anacardium Linn. Int J Pharm Life Sci 2019, 10(1), 6010-2.

There are numerous plants which are used in Ayurveda and other traditional system of medicine but their claim is not yet evaluated scientifically in laboratory Animals. Present study was intended to evaluate an analgesic activity of Semecarpus anacardium fruits extract in mice to confirm ethnomedical claim made by traditional health healer. Dried fruits of plant were extracted in methanol using Soxhlet extractor. Methanolic extract was evaporated using rota evaporator and stored at cool and dry place. Extract was orally administered in mice (200 & 400 mg/kg bw) and evaluated using hot plate and tail immersion methods. Diclofenac Na (50 mg/kg) was taken as a standard drug. Significant analgesic activity comparable with standard dose of diclofenac Na was observed in both the cases. This confirms ethnomedical claim of some workers regarding analgesic activity of the plant.

2 tables, 7 ref

000484   SHINGUMARE B G, MOREGAONKAR S D , GANGANE G R , RAJURKAR S S (Veterinary Pathology Dep, Maharashtra Animal & Fishery Sciences Univ, Maharashtra) : Pathomorphological studies of Acephate induced toxicity in male Wistar rats and its amelioration by Swertia chirata. Int J Chem Sci 2019, 3(1), 19-24.

An experimental trial was conducted on thirty male wistar rats to study acephate induced pathomorphological alterations in male Wistar rats and its amelioration by S. chirata. The study was conducted for twenty eight days and evaluated by recording clinical signs, body weight, relative organ weights and observing gross and histopathological alterations in visceral organs at the end of trial. Group II and group IV male rats showed prominent clinical signs such as increased salivation, piloerection, lethargy, increased urine output, diarrhoea and hair loss (from 14^th day onward), Body weight (from 14^th day onward) and Relative organ weights at 28^th day interval were significantly decreased in a rats of group II and IV. No appreciable gross changes were recorded in organs, except, there was focal areas of necrosis and mild to moderate congestion in liver of rats of group II and IV. Histologically, alterations recorded were haemorrhages, degenerative and necrotic changes in liver and kidney sections; marked depopulation of lymphocytes in spleen; catarrhal enteritis with MNC infiltration in Intestines; depopulation of spermatogonial cells with emptying of seminiferous tubules in Testes; degenerative and necrotic changes in neurons with vacuolations in parenchyma of Brain. These appreciable changes showed improvement due to S. chirata.

11 illus, 3 tables, 22 ref

000455   MARIANNE, HARAHAP U, YUANDANI, FAWZIAH R (Pharmacology Dep, Sumatera Utara Univ, Medan 20155, Indonesia, Email: marianne80@usu.ac.id) : Phytochemical constituent of Picria fel-terrae Lour. ethanol extract and its teratogenic effect. Rasayan J Chem 2019, 12(1), 58 - 63.

The objective of this study is to determine phytochemical constituent in the ethanol extract of Picria fel-terrae leaves as well as to examine the teratogenic effect toward the fetus. There are five groups consists of pregnant rats. Group 1 was the healthy control; groups 2, 3, 4 received extract at the dose of 125, 500 and 1000 mg/kg respectively, group 5 was positive control group (received gabapentin 50 mg/kg). The treatment was performed during the organogenesis period. On the 19th day, maternal laparotomy was carried out, and the fetus was removed. Observations were including reproduction and uterus performance as well as external malformation.Phytochemical screening revealed that the extract consists of flavonoids, tannins, glycosides, saponins and steroids/triterpenoids. Administration extract at the dose of 125, 500, 1000 mg/kg revealed significant effect to the weight loss, fetal body length, number of dead fetuses, resorption and haemorrhages compared to healthy control (p<0.05). The uterus abnormality was found in the group of extract dose of 500 mg/kg as well as 1000 mg/kg. All groups undergo external malformations. The research concluded that all doses of the ethanol extract of P. fel-terrae leaves had the teratogenic effect on the fetus.

2 illus, 2 tables, 23 ref

000444   LEI B, TIAN Z, FAN W, NI B (Microbiology and Immunology Dep, Shanxi Medical Univ, Taiyuan 030001, China, Email: fanweiping26418@126.com) : Circular RNA: A novel biomarker and therapeutic target for human cancers. Int J Med Sci 2019, 16(2), 292-301.

Circular (circ)RNAs, a newly recognized class of noncoding RNA, have been implicated in the occurrence and development of several diseases, including neurological and cardiovascular diseases. Studies of human tumors, including those of liver cancer, gastric cancer, lung cancer and colorectal cancer, have shown differential expression profiles of circRNAs, suggesting regulatory roles in cancer pathogenesis and metastasis. In this review, we discuss the most recent research into tumor-related circRNAs, providing a comprehensive summary of the expression or/and function of these circRNAs and proposing rational perspectives on the potential clinical application of circRNAs as helpful biomarkers or therapeutic targets in human tumors.

2 illus, 1 table, 62 ref

000507   ZHANG P, MING Y, CHENG K, NIU Y, YE Q (Zhongnan Hospital of Wuhan Univ, Hubei- 430 071, China, Email: yqf_china@126.com) : Gene expression profiling in ischemic postconditioning to alleviate mouse liver ischemia/reperfusion injury. Int J Med Sci 2019, 16(2), 343-54.

Ischemic postconditioning (IPO) attenuates hepatic ischemia/reperfusion (I/R) injury. However, little is known about the underlying biological pathophysiology, which could be, at least in part, informed by exploring the transcriptomic changes using next-generation RNA sequencing (RNA-Seq). In this study, 18 mice (C57BL/6) were involved and randomly assigned to three groups: normal (n=6), I/R (n=6, subjected to 70 % hepatic I/R), and IR+IPO (n = 6, applying IPO to mice with I/R injury). We randomly selected 3 mice per group and extracted their liver tissues for next-generation RNA-Seq. We performed a bioinformatics analysis for two comparisons: normal vs. I/R and I/R vs. IR+IPO. From the analysis, 2416 differentially expressed genes (DEGs) were identified (p < 0.05 and fold change ≥ 1.5). Gene ontology (GO) analysis revealed that these genes were mainly related to cellular metabolic processes, nucleic acids and protein binding processes. The enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for the DEGs were the mitogen-activated protein kinase (MAPK), IL-17 signalling pathway, regulating pluripotency of stem cells, and insulin resistance pathway. Validation of 12 selected DEGs by qRT-PCR showed that Cyr61, Atf3, Nr4a1, Gdf15, Osgin1, Egr1, Epha2, Dusp1, Dusp6, Gadd45a and Gadd45b were significantly amplified. Finally, a protein-protein interaction (PPI) network constructed to determine interactions of these 11 DEGs. In summary, by exploring gene expression profiling in regard to hepatic I/R and IPO using next-generation RNA-Seq, we suggested a few progression-related genes and pathways, providing some clues for future experimental research.

7 illus, 2 tables, 49 ref

000491   SUN J, ZHANG J, WANG Y, LI Y, ZHANG R (Hematology Dep, The First Affiliated Hospital of China Medical Univ, Liaoning, 110001, China, Email: hemozerro2008@hotmail.com) : A pilot study of aberrant CpG island hypermethylation of SPRED1 in acute myeloloid leukemia. Int J Med Sci 2019, 16(2), 324-30.

Epigenetic silencing of tumor suppressor genes plays important role in acute myeloid leukemia (AML). Recently, SPRED1, a negative regulator of the RAS MAPK pathway, is identified as a tumour suppressor downregulated in AML. However, little is known regarding its underlying dysregulation in AML. In this study, we investigated methylation status of SPRED1 promoters and their association with mRNA levels in AML. Methylation level were measured in four regions of SPRED1 (#1: 310 bp ~ 723 bp, #2: 810 bp ~ 1299 bp, #3: 1280 bp ~ 1742 bp and #4: 1715 bp ~ 2059 bp) in a total of 16 patients with de novonon-acute promyelocytic leukemia (non-APL) and three patients who got complete remission after induction treatment using the Sequenom MassARRAY platform. Quantitative real-time polymerase chain reaction (q-RT PCR) was used to analyze SPRED1 mRNA levels. AML patients had a significantly higher average methylation level than controls at regions of #1_CpG_1 (p= 0.04) and #1_CpG_11 (p = 0.002). The methylation values for #1_CpG_11 were negatively correlated with mRNA levels (r = -0.558, p = 0.013) but there was no significant association between #1_CpG_1 methylation status and mRNA levels (r = -0.103, p = 0.675) in AML patients. There was no significant difference in the methylation level when comparing with clinical biochemical parameters and treatment response (p > 0.05). Mutations of epigenetic regulation genes such as DNMT3A, TET2 and IDH1/2 were most frequently observed in patients with higher methylation levels. Decreased methylation levels were revealed in three patients who got complete remission. Aberrant methylation statuses of the SPRED1 promoter regions are associated with the downregulation of gene transcription in AML. The methylation level is probably associated with the treatment response of AML. Mutations of epigenetic regulation genes might be involved in the epigenetic aberration of SPRED1.

4 illus, 2 tables, 33 ref

000410   CHYRCHEL B, DROZDZ A, DLUGOSZ D, STEPIEN E L, SURDACKI A (Cardiology Dep, Jagiellonian Univ Medical Coll, PL31-501 Cracow, Poland, Email: andrzej.surdacki@uj.edu.pl) : Platelet reactivity and circulating platelet-derived microvesicles are differently affected by P2Y₁₂ receptor antagonists. Int J Med Sci 2019, 16(2), 264-75.

Platelet-derived microvesicles (PMVs), shed from platelet surface membranes, constitute the majority of circulating microvesicles and have been implicated in procoagulant, pro-inflammatory and pro-atherosclerotic effects. Our aim was to compare plasma PMVs numbers in relation to platelet reactivity during dual antiplatelet therapy (DAPT) with various P2Y₁₂ adenosine diphosphate (ADP) receptor antagonists. In pre-discharge men treated with DAPT for an acute coronary syndrome, plasma PMVs were quantified by flow cytometry on the basis of CD62P (P-selectin) and CD42 (glycoprotein Ib) positivity, putative indices of PMVs release from activated and all platelets, respectively. ADP-induced platelet aggregation was measured by multiple-electrode aggregometry. Clinical characteristics were similar in patients on clopidogrel (n=16), prasugrel (n=10) and ticagrelor (n=12). Platelet reactivity was comparably reduced on ticagrelor or prasugrel versus clopidogrel (p < 0.01). Compared to clopidogrel-treated patients, CD42⁺/CD62P⁺ PMVs counts were 3-4-fold lower in subjects receiving ticagrelor (p=0.001) or prasugrel (p < 0.05), while CD42⁺ PMVs were significantly reduced on ticagrelor (by about 6-fold, p < 0.001), but not prasugrel (p=0.3). CD42⁺/CD62P⁺ PMVs numbers correlated positively to the ADP-induced aggregation on clopidogrel (p < 0.01) or prasugrel (p < 0.05), which was absent in ticagrelor users (p=0.8). CD42⁺ PMVs counts were unrelated to platelet reactivity (p > 0.5). Higher antiplatelet potency of prasugrel and ticagrelor versus clopidogrel is associated with decreased plasma CD42⁺/CD62P⁺ PMVs numbers. However, in contrast to thienopyridines, the association of reduced CD42⁺/CD62P⁺ PMVs counts with ticagrelor use appears independent of its anti-aggregatory effect. Despite similar platelet-inhibitory activity of ticagrelor and prasugrel, only the treatment with ticagrelor seems associated with lower total PMVs release. Our preliminary findings may suggest a novel pleiotropic effect of ticagrelor extending beyond pure anti-aggregatory properties of the drug.

4 illus, 2 tables, 69 ref

000476   ROCA-LEMA D, MARTINEZ-IGLESIAS O, PORTELA C F D A, RODRÍGUEZ-BLANCO A, VALLADARES-AYERBES M, DÍAZ-DÍAZ A, CASAS-PAIS A, PREGO C, FIGUEROA A (Complexo Hospitalario Univ de A Coruña (CHUAC), Sergas, Email: angelica.figueroa.conde-valvis@sergas.es) : In vitro anti-proliferative and anti-invasive effect of polysaccharide-rich extracts from Trametes versicolor and Grifola frondosa in colon cancer cells. Int J Med Sci 2019, 16(2), 231-40.

Colorectal cancer (CRC) is one of leading causes of mortality in western countries and novel treatment strategies are required. The medicinal application of mushrooms has been used in traditional medicine in many oriental countries. Polysaccharide-rich extracts obtained from certain medicinal mushroom species have shown antitumor effects in different experimental models. In the present study, we have developed polysaccharide-rich extracts from Trametes versicolor (TV) and Grifola frondosa (GF) fruit bodies. We aim to evaluate the anticancer effects of these polysaccharide-rich extracts in LoVo and HT-29 human colon cancer cells. The in vitro effects were determined by cytotoxicity assay, proliferation assay, wound healing assay and invasion assay. Moreover, the effect on anchorage independent-cell growth was also determined. Our results showed that TV and GF extracts did inhibit human colon cell proliferation and induce cytotoxicity. Furthermore, both fungal extracts significantly inhibited oncogenic potential, cell migration and invasion in colon cancer cells. In addition, extracts induce a more epithelial phenotype, observed by phase contrast images, together with an increase expression of the E-cadherin epithelial marker, detected by western-blotting analyses. Moreover, by using gelatin zymography assays, it was detected a decrease of MMP-2 enzyme activity, a crucial metalloproteinase important for the degradation of the extracellular matrix. Finally, the combination of the extracts with one the most clinical used agents for colorectal cancer, 5-fluorouracil, increases cell cytotoxicity. Taken together our results underscore a potential antitumor effect of polysaccharide-rich extracts obtained from TV and GF in human colon cancer cells lines. These finding may contribute to the reported health effects of fungal extracts.

5 illus, 1 table, 31 ref

000504   ZAYDMAN A M, STROKOVA E L, STEPANOVA A O, LAKTIONov P P, SHEVCHENKO A I, SUBBOTIN V M (Novosibirsk Research Institute of Traumatology and Orthopaedics, Novosibirsk, Russia, Email: AZaydman@niito.ru) : A new look at causal factors of idiopathic scoliosis: Altered expression of genes controlling chondroitin sulfate sulfation and corresponding changes in protein synthesis in vertebral body growth plates. Int J Med Sci 2019, 16(2), 221-30.

In a previous report, we demonstrated the presence of cells with a neural/glial phenotype on the concave side of the vertebral body growth plate in Idiopathic Scoliosis (IS) and proposed this phenotype alteration as the main etiological factor of IS. In the present study, we utilized the same specimens of vertebral body growth plates removed during surgery for Grade III–IV IS to analyse gene expression. We suggested that phenotype changes observed on the concave side of the vertebral body growth plate can be associated with altered expression of particular genes, which in turn compromise mechanical properties of the concave side. We used a Real-Time SYBR Green PCR assay to investigate gene expression in vertebral body growth plates removed during surgery for Grade III–IV IS; cartilage tissues from human fetal spine were used as a surrogate control. Special attention was given to genes responsible for growth regulation, chondrocyte differentiation, matrix synthesis, sulfation and transmembrane transport of sulfates. We performed morphological, histochemical, biochemical, and ultrastructural analysis of vertebral body growth plates. Expression of genes that control chondroitin sulfate sulfation and corresponding protein synthesis was significantly lower in scoliotic specimens compared to controls. Biochemical analysis showed 1) a decrease in diffused proteoglycans in the total pool of proteoglycans; 2) a reduced level of their sulfation; 3) a reduction in the amount of chondroitin sulfate coinciding with raising the amount of keratan sulfate; and 4) reduced levels of sulfation on the concave side of the scoliotic deformity. The results suggested that altered expression of genes that control chondroitin sulfate sulfation and corresponding changes in protein synthesis on the concave side of vertebral body growth plates could be causal agents of the scoliotic deformity.

4 illus, 2 tables, 49 ref

000431   HUANG Y-H, KUO H-C, YANG Y-L, WANG F-S (Anesthesiology Dep, Chang Gung Univ, Kaohsiung, Taiwan, Email: wangfs@ms33.hinet.net) : MicroRNA-29a is a key regulon that regulates BRD4 and mitigates liver fibrosis in mice by inhibiting hepatic stellate cell activation. Int J Med Sci 2019, 16(2), 212-20.

MicroRNA-29a is a key regulon that regulates hepatic stellate cells (HSCs) and mitigates liver fibrosis. However, the mechanism by which it does so remains largely undefined. The inhibition of bromodomain-4 protein (BRD4) represents a novel therapeutic target in hepatic fibrosis. Therefore, the purpose of this study is to investigate the miR-29a regulation of BRD4 signaling in a bile duct-ligation (BDL) animal model with regard to developing cholestatic liver fibrosis. Hepatic tissue in miR-29a transgenic mice (miR-29aTg mice) displayed weak fibrotic matrix, as shown by α-smooth muscle actin staining within affected tissues compared to wild-type mice. miR-29a overexpression reduced the BDL exaggeration of BRD4 and SNAI1 expression. Increased miR-29a signaling caused the downregulation of EZH2, MeCP2, and SNAI1, as well as the upregulation of PPAR-γ expression, in primary HSCs. We further demonstrated that the administration of JQ1, a BRD4 inhibitor, could inhibit BRD4, C-MYC, EZH2, and SNAI1 expression, while both JQ1 and a miR-29a mimic could inhibit the migration and proliferation of HSCs. In short, our research demonstrates that miR-29a negatively regulates HSC activation by inhibiting BRD4 and EZH2 function, thus making it a promising target for the pharmacologic treatment of hepatic fibrosis.

6 illus, 38 ref

000506   ZHANG M, SRICHAI M B, ZHAO M, CHEN J, DEVIS L S, WU G, BREYER M D, HAO C-M (Nephrology Div, Fudan Univ, Shanghai, China, Email: chuanminghao@fudan.edu.cn) : Nonselective cyclooxygenase inhibition retards cyst progression in a murine model of autosomal dominant polycystic kidney disease. Int J Med Sci 2019, 16(1), 180-8.

Autosomal dominant polycystic kidney disease is one of the most common genetic renal diseases. Cyclooxygenase plays an important role in epithelial cell proliferation and may contribute to the mechanisms underlying cyst formation. The aim of the present study was to evaluate the role of cyclooxygenase inhibition in the cyst progression in polycystic kidney disease. Pkd2^WS25/- mice, a murine model which harbors a compound cis-heterozygous mutation of the Pkd2 gene were used. Cyclooxygenase expression was assessed in both human and murine kidney specimens. Pkd2^WS25/- mice were treated with Sulindac (a nonselective cyclooxygenase inhibitor) or vehicle for 8 months starting at three weeks age, and then renal cyst burden was assessed by kidney weight and volume. Cyclooxygenase-2 expression was up-regulated compared to control kidneys as shown by RNase protection in human polycystic kidneys and immunoblot in mouse Pkd2^WS25/- kidneys. Cyclooxygenase-2 expression was up-regulated in the renal interstitium as well as focal areas of the cystic epithelium (p<0.05). Basal Cyclooxygenase-1 levels were unchanged in both immunohistochemistry and real-time PCR. Administration of Sulindac to Pkd2^WS25/- mice and to control mice for 8 months resulted in reduced kidney weights and volume in cystic mice. Renal function and electrolytes were not significantly different between groups. Thus treatment of a murine model of polycystic kidney disease with Sulindac results in decreased kidney cyst burden. These findings provide additional implications for the use of Cyclooxygenase inhibition as treatment to slow the progression of cyst burden in patients with polycystic kidney disease.

6 illus, 1 table, 35 ref

000407   CHANG C-C, SIA K-C, CHENG J-F, LIN C-M, YANG C-M, HUANG K-Y, LIN W-N (Fu Jen Catholic Univ, New Taipei City 242, Taiwan, Email: 081551@mail.fju.edu.tw) : Lipopolysaccharide promoted proliferation and adipogenesis of preadipocytes through JAK/STAT and AMPK-regulated cPLA2 expression. Int J Med Sci 2019, 16(1), 167-79.

The proliferation and adipogenesis of preadipocytes played important roles in the development of adipose tissue and contributed much to the processes of obesity. On the other hand, lipopolysaccharide (LPS), also known as endotoxin, is a key outer membrane component of gram-negative bacteria in the gut microbiota, and has a dominant role in linking inflammation to high-fat diet-induced metabolic syndrome. Studies suggested the potential roles of LPS in hepatic steatosis and in obese mice models. However, the molecular mechanisms underlying LPS-regulated obesity remained largely unknown. Here we reported that LPS stimulated expression of cyosolic phospholipase A2 (cPLA2), one of inflammation regulators of obesity, in the preadipocytes. Pretreatment the inhibitors of JAK2, STAT3, STAT5 or AMPK significantly reduced LPS-increased mRNA and protein expression of cPLA2 together with phosphorylation of JAK2, STAT3, STAT5 and AMPK, separately. Similarly, transfection of siRNA against JAK2 or AMPK abolished expression of cPLA2 and phosphorylation of JAK2 or AMPK together with downregulated expression of JAK2 and AMPK protein. LPS enhanced activation of STAT3 and STAT5 via JAK2-dependent manner in the preadipocytes. Transfection of JAK2 or AMPK siRNA further proofed the independence of JAK2 and AMPK in LPS-treated preadipocytes. In addition, LPS-increased DNA synthesis, cell numbers and cell viability of preadipocytes were attenuated by AACOCF3, AG490, BML-275, cPLA2 siRNA, JAK2 siRNA or AMPK siRNA. Attenuation JAK2/STAT or AMPK-dependent cPLA2 expression reduced LPS-mediated adipogenesis of preadipocytes. Stimulation of arachidonic acid or AMPK activator, A-769662, increased cell numbers and cell viability and promoted differentiation of preadipocytes. Collectively, these results indicated that LPS increased preadipocytes proliferation and adipogenesis via JAK/STAT and AMPK-dependent cPLA2 expression. The mechanisms of LPS-stimulated cPLA2 expression may be a link between bacteria and obesity and provides the molecular basis for preventing metabolic syndrome or hyperplasic obesity.

8 illus, 51 ref

000434   JEONG J Y, CHA H-J, CHOI E O, KIM C H, KIM G-Y, YOO Y H, HUANG H-J, PARK H T, YOON H M, CHOI Y H (Physiology Dep, Dong-A Univ, Busan 49201, Republic of Korea, Email: phwantae@dau.ac.kr) : Activation of the Nrf2/HO-1 signaling pathway contributes to the protective effects of baicalein against oxidative stress-induced DNA damage and apoptosis in HEI193 Schwann cells. Int J Med Sci 2019, 16(1), 145-55.

Baicalein, a flavonoid extracted from the roots of Scutellaria baicalensis Georgi., has various pharmacological effects due to its high antioxidant activity. However, no study has yet been conducted on the protective efficacy of baicalein against oxidative stress in Schwann cells. In this study, we evaluated the protective effect of baicalein on DNA damage and apoptosis induced by hydrogen peroxide (H₂O₂) in HEI193 Schwann cells. For this purpose, HEI193 cells exposed to H₂O₂ in the presence or absence of baicalein were applied to cell viability assay, immunoblotting, Nrf2-specific small interfering RNA (siRNA) transfection, comet assay, and flow cytometry analyses. Our results showed that baicalein effectively inhibited H₂O₂-induced cytotoxicity and DNA damage associated with the inhibition of reactive oxygen species (ROS) accumulation. Baicalein also weakened H₂O₂-induced mitochondrial dysfunction, increased the Bax/Bcl-2 ratio, activated caspase-9 and -3, and degraded poly(ADP-ribose) polymerase. In addition, baicalein increased not only the expression but also the phosphorylation of nuclear factor-erythroid 2 related factor 2 (Nrf2) and promoted the expression of heme oxygenase-1 (HO-1), a critical target enzyme of Nrf2, although the expression of kelch-like ECH-associated protein-1 was decreased. However, the inhibition of Nrf2 expression by transfection with Nrf2-siRNA transfection abolished the expression of HO-1 and antioxidant potential of baicalein. These results demonstrate that baicalein attenuated H₂O₂-induced apoptosis through the conservation of mitochondrial function while eliminating ROS in HEI193 Schwann cells, and the antioxidant efficacy of baicalein implies at least a Nrf2/HO-1 signaling pathway-dependent mechanism. Therefore, it is suggested that baicalein may have a beneficial effect on the prevention and treatment of peripheral neuropathy induced by oxidative stress.

6 illus, 44 ref

000422   GUNDAMARAJU R, VEMURI R, CHONG W C, BULMER A C, ERI R (Tasmania Univ, Tasmania, Australia, Email: rohit.gundamaraju@utas.edu.au) : Bilirubin attenuates ER stress-mediated inflammation, escalates apoptosis and reduces proliferation in the LS174T colonic epithelial cell line. Int J Med Sci 2019, 16(1), 135-44.

Mildly elevated serum unconjugated bilirubin (UCB) concentrations are associated with protection against disease conditions underpinned by cellular and metabolic stress. To determine the potential therapeutic efficacy of UCB we tested it in an in vitro model of gut inflammation. Tunicamycin TUN (10 µg/mL) was used to induce endoplasmic reticular stress (ERS) affecting N-glycosylation in LS174T cells. Cultured cells were investigated with addition of UCB at doses 0.1, 1 and 10 µM (resulting in bilirubin: albumin ratios of 0.325–0.003) against ER stress-mediated effects including inflammation, cell survival (determined by apoptosis) and proliferation. Gene expression of ER stress markers (Grp78, Perk, XBP1 and ATF6) were evaluated in addition to cytokine concentrations in media after six hours of treatment. We then verified the potential role of UCB in executing programmed cell death via PARP, Caspase3 and Annexin V assays and further explored cell proliferation using the Click-iT EdU assay. A dose of 10 µM UCB most potently reduced tunicamycin-mediated effects on enhanced UPR markers, inflammatory cytokines and proliferation; however all the doses (i.e.0.1–10 µM) reduced the expression of ER stress and inflammatory markers Grp78, NLRP3, IL1-b, XBP1, PERK and ATF6. Furthermore, media concentrations of pro-inflammatory cytokines IL-8, IL-4 and TNFα decreased and the anti-inflammatory cytokine IL-10 increased (P<0.05). A dose of 10 µM UCB initiated intrinsic apoptosis via Caspase 3 and in addition reduced cellular proliferation. Collectively, these data indicate that co treatment with UCB resulted in reducing ER stress response to TUN in gastrointestinal epithelial cells, reduced the subsequent inflammatory response, induced cancer cell death and decreased cellular proliferation. These data suggest that mildly elevated circulating or enteric UCB might protect against gastrointestinal inflammatory disorders.

6 illus, 55 ref

000490   SUH S-S, HONG J-M, KIM E J, JUNG S W, CHAE H, KIM J E, KIM J H, KIM I-C, KIM S (Polar Life Sciences Div, Korea Polar Research Institute, Incheon, 21990, Republic of Korea, Email: sangheekim@kopri.re.kr) : Antarctic freshwater microalga, Chloromonas reticulata, suppresses inflammation and carcinogenesis. Int J Med Sci 2019, 16(1), 189-97.

Inflammation triggered by the innate immune system is a strategy to protect organisms from the risk of environmental infection. However, it has recently become clear that inflammation can cause a variety of human diseases, including cancer. In this study, we investigated the effects of an ethanol extract of the Antarctic freshwater microalgae, Chloromonas reticulata (ETCH), on inflammation and carcinogenesis in RAW 264.7 macrophages and HCT116 human colon cancer cells, respectively. ETCH exhibited significant anti-inflammatory activity through the dose-dependent modulation of major inflammatory markers such as COX-2, IL-6, iNOS, TNF-α, and NO production. For example, ETCH reduced LPS-induced upregulation of COX-2, IL-6, iNOS, and TNF- alpha mRNA levels, leading to a significant decrease in the levels of LPS-stimulated NO and IL-6 as well as TNF-alpha products. In contract, ETCH exhibited dose-dependent cytotoxic activity against HCT116 cells, yielding a profound reduction in the proliferation of the cancer cells. Furthermore, ETCH induced G2 phase cell cycle arrest by transcriptionally regulating of genes involved in G2 / M transition including p21 (CDKN1A), cyclin B1 (CCNB1), and CDK1; CDKN1A mRNA levels were upregulated in response to ETCH, whereas CCNB1 and CDK1 were downregulated. This study reports for the first time anti-inflammatory and anti-cancer effects of, C. reticulata and provides new insights into the molecular mechanisms of the linkage between inflammation and cancer.

6 illus, 30 ref

000469   PIRCHER A, SCHÄFER G, EIGENTLER A, PICHLER R, PUHR M, STEINER E, HORNINGER W, GUNSILIUS E, KLOCKER H, HEIDEGGER I (Urology Dep, Medical Univ, Innsbruck, Austria, Email: Isabel-maria.heidegger@i-med.ac.at) : Robo 4 - the double-edged sword in prostate cancer: Impact on cancer cell aggressiveness and tumor vasculature. Int J Med Sci 2019, 16(1), 115-24.

The magic roundabout receptor 4 (Robo 4) is a tumor endothelial marker expressed in the vascular network of various tumor entities. However, the role of Robo 4 in prostate cancer (PCa), the second common cause of cancer death among men in –developed countries, has not been described yet. Thus, the present study investigates for the first time the impact of Robo 4 in PCa both in the clinical setting and in vitro. Immunohistochemical analyses of benign and malignant prostate tissue samples of 95 PCa patients, who underwent radical prostatectomy (RPE), revealed a significant elevated expression of Robo 4 as well as its ligand Slit 2 protein in cancerous tissue compared to benign. Moreover, increased Robo 4 expression was associated with higher Gleason score and pT stage. In advanced stage we observed a hypothesis-generating trend that high Robo 4 and Slit 2 expression is associated with delayed development of tumor recurrence compared to patients with low Robo 4 and Slit 2 expression, respectively. In contrast to so far described exclusive expression of Robo 4 in the tumor vascular network, our analyses showed that in PCa Robo 4 is not only expressed in the tumor stroma but also in cancer epithelial cells. This finding was also confirmed in vitro as PC3 PCa cells express Robo 4 on mRNA as well as protein level. Overexpression of Robo 4 in PC3 as well as in Robo 4 negative DU145 and LNCaP PCa cells was associated with a significant decrease in cell-proliferation and cell-viability. In summary we observed that Robo 4 plays a considerable role in PCa development as it is expressed in cancer epithelial cells as well as in the surrounding tumor stroma. Moreover, higher histological tumor grade was associated with increased Robo 4 expression; controversially patients with high Robo 4 tend to exert lower biochemical recurrence possibly reflecting a protective role of Robo 4.

7 illus, 2 tables, 33 ref

000447   LI H, ZHAO J, LIU B, LUO J, LI Z, QIN X, WEI Y (Cardiology Dep, Tongji Univ School of Medicine, Shanghai, People’s Republic of China, Email: ywei@tongji.edu.cn.) : MicroRNA-320 targeting neuropilin 1 inhibits proliferation and migration of vascular smooth muscle cells and neointimal formation. Int J Med Sci 2019, 16(1), 106-14.

This study shows that microRNA-320 (miR-320) is associated with many important cell functions, including cell differentiation, proliferation, migration, and apoptosis. However, the role of miR-320 in vascular smooth muscle cells (VSMCs) and proliferative vascular diseases is still completely unclear. In our study, we found that the expression of miR-320 in human VSMCs after PDGF stimulation was significantly down-regulated in time- and dose-dependent manner. Function analyses identified that miR-320 could inhibit the proliferation and migration of VSMCs in both basal and PDGF-stimulated conditions. Furthermore, Neuropilin 1 (NRP1) was demonstrated as a direct target of miR-320 in Luciferase reporter assays and miR-320 overexpression inhibited the expression of NRP1 with or without PDGF treatment. Finally, miR-320 was markedly decreased in mice carotid arteries after ligated injury, while the restoration of miR-320 via Ad-miR-320 attenuated neointimal hyperplasia by declining the NRP1 expression. The results confirmed that miR-320 regulated proliferation and migration of VSMCs and neointimal formation by targeting NRP1. These novel findings implied that the regulation of NRP1 expression by miR-320 has important significance in the early diagnosis and treatment of proliferation vascular diseases.

6 illus, 30 ref

000436   JIANG H, YU Y, LIU S, ZHU M, DONG X, WU J, ZHANG Z, ZHANG M, ZHANG Y (Neurology and Neuroscience Center Dep, First Hospital of Jilin Univ, Changchun, Jilin Province, China, Email: rose19700@sina.com) : Proteomic study of a Parkinson’s disease model of undifferentiated SH-SY5Y cells induced by a proteasome inhibitor. Int J Med Sci 2019, 16(1), 84-92.

Parkinson's disease (PD) is one of the most common nervous system degenerative diseases. However, the etiology of this disease remains elusive. Here, a proteasome inhibitor (PSI)-induced undifferentiated SH-SY5Y PD model was established to analyze protein alterations through proteomic study. Cultured undifferentiated SH-SY5Y cells were divided into a control group and a group treated with 2.5 µM PSI (PSI-treated group). An methyl thiazolyl tetrazolium (MTT) assay was applied to detect cell viability. Acridine orange/ethidium bromide (AO/EB), α-synuclein immunofluorescence and hematoxylin and eosin (H&E) staining were applied to evaluate apoptosis and cytoplasmic inclusions, respectively. The protein spots that were significantly changed were separated, analyzed by 2D gel electrophoresis and DIGE De Cyder software, and subsequently identified by MALDI-TOF mass spectrometry and database searching. The results of the MTT assay showed that there was a time and dose dependent change in cell viability following incubation with PSI. After 24 h incubation, PSI resulted in early apoptosis, and cytoplasmic inclusions were found in the PSI-treated group through H&E staining and α-synuclein immunofluorescence. Thus, undifferentiated SH-SY5Y cells could be used as PD model following PSI-induced inhibition of proteasomal function. In total, 18 proteins were differentially expressed between the groups, 7 of which were up-regulated and 11 of which were down-regulated. Among them, 5 protein spots were identified as being involved in the ubiquitin proteasome pathway-induced PD process. Mitochondrial heat shock protein 75 (MTHSP75), phosphoglycerate dehydrogenase (PHGDH), laminin binding protein (LBP), tyrosine 3/tryptophan 5-monooxygenase activation protein (14-3-3ε) and YWHAZ protein (14-3-3ζ) are involved in mitochondrial dysfunction, serine synthesis, amyloid clearance, apoptosis process and neuroprotection. These findings may provide new clues to deepen our understanding of PD pathogenesis.

5 illus, 2 tables, 30 ref

000492   SUN L, YUAN H, BURNETT J, GASPARYAN M, ZHANG Y, ZHANG F, YANG Z, RAN Y, SUN D (Peking Union Medical Coll, Beijing, 100021, P. R. China, Email: sunlichao@cicams.ac.cn) : MEOX1 promotes tumor progression and predicts poor prognosis in human non-small-cell lung cancer. Int J Med Sci 2019, 16(1), 68-74.

MEOX1 is a homeobox transcriptional factor, and plays essential roles in regulating somite development. Our previous study indicated that MEOX1 is a critical molecular target in mesenchymal-like cancer cells in PTEN-deficient Trastuzumab resistant breast cancer. Despite the potential implication of MEOX1 for the cancer progression, no previous studies examined its level and clinical significance in lung cancer tissues. In this study, we aimed to detect the MEOX1 expression and correlate its level with clinical outcome in non-small-cell lung cancer patients (NSCLC). MEOX1 gene expression in lung cancer was examined by using the Oncomine database. MEOX1 protein levels were evaluated by IHC using the corresponding primary antibody on two different commercial lung cancer tissue arrays. siRNA knockdown was used to elucidate the function of MEOX1. Analysis of the Oncomine datasets identified that an elevation of MEOX1 in gene amplification in lung cancer tissues in comparison to normal lung tissues. Immunohistochemistical analysis demonstrated that MEOX1 was localized predominantly in the nucleus, and positive rate was 67.3 % (111/165) in NSCLC samples. Statistical analysis revealed high levels of MEOX1 significantly correlated with Lymph Node Metastasis and Stage. Kaplan-Meier survival analysis showed that high levels of MEOX1 were significantly associated with unfavorable survival in NSCLC patients, and MEOX1 nucleus staining had worse survival, than did patients with overall expression in lung squamous cell carcinoma patients. Multivariate Cox's regression analysis found that MEOX1 was an independent poor prognostic predictor for patients with NSCLC. Silencing of MEOX1 by specific SiRNA significantly inhibited H460 and H1299 cell proliferation and sphere formation in serum-free medium. Our results firstly indentified that high levels of MEOX1 especially nuclear staining was an independent prognostic factor for NSCLC, and it served a essential roles in the regulation of cell proliferation and colony formation in vitro. It may represent a potential target for the NSCLC treatment.

4 illus, 2 tables, 15 ref

000500   WANG X , LAI Q, HE J, LI Q, DING J, LAN Z, GU C, YAN Q, FANG Y, ZHAO X, LIU S (Gastroenterology Dep, Southern Medical Univ, Guangzhou, People’s Republic of China, Email: liuside2011@163.com) : LncRNA SNHG6 promotes proliferation, invasion and migration in colorectal cancer cells by activating TGF-β/Smad signaling pathway via targeting UPF1 and inducing EMT via regulation of ZEB1. Int J Med Sci 2019, 16(1), 51-59.

Long noncoding RNAs (lncRNAs) are non-protein coding transcripts longer than 200 nucleotides in length. They drive many important cancer phenotypes through their interactions with other cellular macromolecules including DNA, RNA and protein. Recent studies have identified numerous lncRNAs active in colorectal cancer (CRC). The lncRNA small nucleolar RNA host gene 6 (SNHG6) has been reported to have an oncogenic role in multiple cancers. However, the biological role and mechanism of SNHG6 in the tumorigenesis of CRC has not been reported in-deep. The Cancer Genome Atlas (TCGA) database and GEO database were used to identify SNHG6 expression in different human cancers and explore the relationship between SNHG6 expression and patient prognosis using Kaplan-Meier method analysis. SNHG6 expression in 77 pairs of clinical CRC tissues and different CRC cell lines were analyzed by quantitative real-time PCR (qRT-PCR). A CCK-8 assay was used to assess cell proliferation, transwell assay to detect the cell metastasis, and tumor growth was investigated with a nude mice model in vivo. Whether UPF1 and ZEB1 are downstream targets of SNHG6 was verified by bioinformatics target gene prediction, qRT-PCR and western blot. TCGA data showed that SNHG6 was significantly upregulated in colorectal cancer samples in comparison with healthy data samples (P < 0.01). CRC patients with high levels of SNHG6 had a significantly shorter overall survival than those with low levels of SNHG6 (P = 0.0162). qRT-PCR confirmed that the expression of SNHG6 was significantly upregulated in CRC tissues and cell lines. Upregulation of SNHG6 expression induced RKO and HCT116 cell proliferation as well as RKO cell metastasis, while downregulation of SNHG6 expression supressed the proliferation and metastasis of RKO cells and tumor growth in vivo. UPF1 was upregulated and ZEB1 was decreased when SNHG6 knockdown, regulating the TGF-β/Smad pathway and inducing EMT respectively. SNHG6 may play an oncogenic role in CRC cells by activating TGF-β/Smad signaling pathway via targeting of UPF1 and inducing EMT via regulating ZEB1. This could be a prognostic biomarker and therapeutic target for CRC.

6 illus, 2 tables, 37 ref