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Correlation between interleukin-21 and formation of intrauterine adhesions in rats

Huihua CAI ; Yuanli HE ; Huijuan LI ; Jingxiong LI ; Fang LIU

Journal of Medical Postgraduates.2015;(4):346-349.

Objective Interleukin-21(IL-21) is likely to contribute to the development of liver fibrosis, but up to now, no study has been reported on the relationship between IL-21 and intrauterine adhesions ( IUA) .This study aimed to establish a rat model of IUA induced by mechanical injury and lipopolysaccharide (LPS) infection (dual injury), determine the expression level of serum IL-21, and confirm the association of serum IL-21 with the formation of IUA. Methods Forty healthy female SD rats were randomly divided into four groups of equal number,control, mechanical injury, LPSinfection, and dual injury.At 7 days after IUA modeling,uterine tissue-swere collected from all the animals for observation of the endometrial glands,detection of the degree of IUA by Masson staining, measure-ment of the serum IL-21 level by radioimmunoassay, and analysis of the correlation between the number of endometrial glands and the de-gree of fibrosis. Results The number of endometrial glandswas significantly smallerin the dual injury group (3.59±1.20) than in the mechanical injury (11.66±2.34) and LPSinfection group(11.59±1.47)(P<0.05), while the proportion of fibrosis area wassignificantly higher in the former group(0.65±0.03) than in the lattertwo(0.30±0.07 and 0.32±0.08)(P<0.05).The level of serum IL-21 was signifi-cantly increased in the dual injury group ([286.21±27.80]pg/mL) as compared with those in the control ( [ 118.65 ±22.55 ] pg/mL ) , mechanical injury([176.20±19.05]pg/mL), and LPS infection group ([187.98±16.51]pg/mL) (P<0.05), with a positive correlation be-tween the IL-21 level and theproportion of fibrosis area ( r=0.271, P<0.05) . Conclusion A rat model ofIUAwas successfully established by mechanical injury and lipopolysaccharide (LPS) infection.The evi-dent increase of serum IL-21 in the IUA model was positively correlated with the percentage of fibrosis area, suggesting that IL-21 may be involved indirectly in the formation of IUA.

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Effects of metformin on cell proliferation and fatty acid synthase in human hepatocellular carcinoma cell line HepG2

Xiaoren PENG ; Yan LIU ; Dajin ZOU ; Juan LI

Journal of Medical Postgraduates.2015;(4):360-364.

Objective The cancer risk of patients with diabetes mellitus who are treated by metformin declines remarkably in comparison to patients receiving other drug therapies.The article was to investigate the relationship between antineopastic activity and fatty acid synthase (FASN) of metformin in human hepatocellular carcinoma cell(HCC) line HepG2. Methods HepG2 cells were treated with various concentrations of metformin( 0, 1, 5, 10, 15 mmol/L) for 24, 48 and 72 h respectively and cell growth was assessed by CCK-8 assay.Positive control(paclitaxel 10μg/mL) and negative control(metformin 0mmol/L) were set up simultaneously.After being treated with doses of metformin(0, 5, 10,15mmol/L) for 72h, protein expression levels of AMPKα、P-AMPKα、FASN、P-mTOR and P-Akt were measured by western blotting analysis and FASN mRNA expression levels were measured by RT-PCR. Results Being treated with vari-ous doses of metformin(1, 5, 10, 15 mmol/L) for 24, 48 and 72 h, the growth of HepG2 cells were inhibited by metformin in dose-dependent and time-dependent manner( P<0.05) .The growth inhibition rate approached 50%after being treated with metformin for 72 h, and the growth inhibition rate were all greater than 50%after being treated with 15 mmol/L of metformin.At 72 h after the treatment of various do-ses of metformin(0, 5, 10, 15 mmol/L) on HepG2 cells, the protein expression of P-AMPK increased with the rise of metformin concentra-tion, while the protein expressions of P-mTOR and FASN decreased as the metformin concentration increased.Compared with negative control group, the protein expression levels of P-AMPKα, P-mTOR and FASN all changed significantly in 10 mmol/L group and 15 mmol/L group(P<0.05).However, there was no significant difference as to the protein expression levels of P-Akt in various metformin concen-trations( P>0.05) .FASN mRNA expression levels decreased significantly in all metformin-treated groups( P<0.05) . Conclusion Met-formin actitiviates AMPK, inhibits mTOR and downregulates FASN, which are implicated in its antineopastic activity on HCC.Although metformin inhibits mTOR activation, it is not involved in Akt upregulation through a negative loop.

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Construction of a C3aR expres sion vector and infection in tubular epithelial cells

Jingmin ZHENG ; Guang YIN ; Wenjin ZHAO ; Zhihong LIU

Journal of Medical Postgraduates.2015;(4):355-359.

Objective Our previous study showed that the expression of C 3aR was increased in renal tubular epithelial cells (RTEC).However, the role of C3aR in diabetic nephropathy remains unknown, and so does the exact physiological and pathological signifi-cance of C3aR in relevant renal tissues.In the present study, we investigated the physiological and pathological significance of C3aR in RTECs using a lentivirus expression vector and anRTEC strain overexpressing C3aR. Methods Based on the sequence of C3aR mRNA, the human C3aR gene was synthesized and cloned into the multi-clonal site of the lentivirus expression vector pLenti6.3-MCS-IRES2-EGFP to construct a C3aR expression vector pLenti6.3-C3aR-IRES2-EGFP.After identification by sequencing, the recombinant lentivirus expressing C3aR was packaged by cotransfecting293T cellswith the recombinant lentivirusexpression vector and packaged plasmid .Then,the recombinant lentivirus was used to infect the human RTECline HK 2.After screening in the medium with blasticidin, blasticidinr-esistant cell clones were obtained, followed by identification ofthe human RTECline stably overexpressing C 3aR by real-time PCR and immunochemical staining. Results TheC3aR expression vector pLenti6.3-C3aR-IRES2-EGFP was successfully constructed and the sequence was proved to be correct. C3aR expression recombinant lentivirus was successfully packaged with a titer of 5×108/mL.Blasticidin-resistant HK2 cell clones were ob-tained and the expression of HK2-C3aR mRNA was significantly higherin the HK2-C3aR cells than in the non-transfected HK2 cells (2.33± 0.45 sv 1.00±0.09, P<0.05). Conclusion We successfully constructed a C3aR expression lentivirus vector pLenti6.3-C3aR-IRES 2-EGFP and a C3aR overexpression renal tubular epithelial cell strain HK2-C3aR, which is very contributive to further studies of the roles of C3aR in renal tubular epithelial cells and other types of cells.

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Effects of the F10 gene overexpression and silence on the expressions of invasion-related proteinases in choriocarcinoma cell line JAR

Xiaohua SU ; Zhanjun PANG

Journal of Medical Postgraduates.2015;(4):350-354.

Objective The F10 gene was found in the initial stage of our study to be highly expressed in hydatidiform mole. The aim of this study was to investigate the effects of theoverexpression or depletion of F10 on the invasiveness of the choriocarcinoma cell line JAR and explore the relationship of F10 expression with the invasiveness and metastasis ofchoriocarcinoma. Methods Using cell transfection and RNA interference technology, we established JAR choriocarcinoma cell lines with stablyoverexpressedor silenced F10 gene.We randomly and equally assigned 30 SPF nude mice into the three groups to receive the injection of JAR cellswith overex-pressed F10 ( F10 overexpression group) , untreated JAR cells ( control group) , and JAR cells with silenced F10( F10 silence group) . At 5 weeks after the JAR cell injection, we harvested the subcutaneous tumor tissues from the mice, determined the expressions of ma-trix metalloproteinases (MMP), tissue inhibitors of metalloproteinase-1(TIMP-1), and plasminogen activator inhibitor-1(PAI-1), and compared by Western blot and immunohistochemistry, and compared the expressions among the three groups of mice. Results Im-munohistochemistryshowed significantlyup-regulated expressions of MMP-2, -8, -11, -16, and-19 and down-regulated expressions of TIMP-1 and PAI-1 in the subcutaneous tumor tissues in the F10 overexpression group as compared with the control and F10 silence groups ( P<0.05) .Western blot also exhibitedextremely significantly up-regulated expressions of MMP-2,-8,-11,-16, and -19 pro-teins anddown-regulated expressions of TIMP-1 and PAI-1 proteins in the F10 overexpression groupin comparison with the control and F10 silence groups( P<0.01 ) . Conclusion By up-regulating the expressions of MMPs and down-regulating the expressions of TIMP-1 and PAI-1, the F10 gene might be an upstream stimulating factor involved in the proliferation, invasiveness, and metastasis of-choriocarcinoma cells.

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Research progress of drugs in treatment of stress gastric ulcer

Zhikun LI ; Fuwen WANG

Journal of Medical Postgraduates.2015;(1):110-112.

The stress gastric ulcer is due to a stress-induced gastric erosions, superficial ulcers and bleeding gastric mucosal le-sions characterized by stress. Its etiology is closely associated with lives, the incidence is increasing. In recent years, the the basic re-search of drugs focuses on inhibition of oxidative damage caused by stress, anti-inflammatory cytokine expression, and regulation of gastric acid, etc., and made a lot of success. Treatment choice of stress ulcer drug gradually increased. The effects of plant extracts on stress ul-cer get more and more attention. This article reviews research progress of the treatment drugs of stress ulcer in the current stage.

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Inhibition of human mda-7/IL-24 gene on the proliferation of human lung cancer cells

Qunfeng MA

Journal of Medical Postgraduates.2003;0(09):-.

Objective:To analyze the role of human mda-7/IL-24 gene on the proliferation of human non-small cell lung cancer cell line (H460). Methods:The mda-7/IL-24 cDNA was subcloned into expression vector pcDNA3 and transient transfected in human lung carcinoma cell H460. The overexpressed mda-7/IL-24 was identified by RT-PCR using SP6 primer and mda-7/IL-24 upstream primer. The role of human mda-7/IL-24 on the proliferation of H460 cells was analyzed with MTT after transfection of (pcDNA3)-mda-7/IL-24 for 72h. Results:The mda-7/IL-24 cDNA was subcloned into pcDNA3 successfully. RT-PCR study showed that mda-7/IL-24 could express in H460 cells at different ratio of DNA and lipofectamine (1 ?g ∶ 2?l-1?g ∶ 4?l). Compared with H460 cells and H460 cells transfected with (pcDNA3), pcDNA3-mda -7/IL-24 could inhibit the proliferation of H460 cells. The inhibition rate was (18.4%). Conclusion:The mda-7/IL-24 can inhibit the proliferation of human non-small-cell lung carcinoma cell H460.

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Impact of dehydroepiandrosterone on antioxidant capacity of Raji cell cultured in vitro

Detai ZHANG

Journal of Medical Postgraduates.2003;0(09):-.

Objective: To investigate the influence of dehydroepiandrosterone(DHEA) on antioxidant capacity of Raji cell cultured in vitro. Methods: Raji cell line was cultured in vitro in the presence of DHEA and/or phenazine methosulfate(PMS), some antioxidant enzymes including glucose-6-phosphate dehydrogenase activity(G-6-PD),glutathione peroxidase activity(GPX), glutathione reductase activity(GR) and intracellular glutathione(GSH) concentration of the DHEA pretreatment cells were evaluated. meanwhile, these results were compared with that from the respective control cells. Results:(G-6-PD) activities were suppressed significantly and no changes in activities of GPX,GR and intracellular GSH concentration were occurred while Raji cells were cultured in the presence of DHEA at concentration more than 5.0?mol/L . On the other hand, the DHEA pretreatment cells show no changes in (G-6-PD,)GPX,GR activities and significantly decreased GSH concentration(P

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Integrated backscatter of kidney in rabbits with different dosage poisoning

Junying CAO

Journal of Medical Postgraduates.2003;0(10):-.

Objective: To explore the value of integrated backscatter(IBS) of kidney with different dosage of organophosphorus in rabbits. Methods: Animal models of organophosphorus poisoning were established. Ultrasonographic examinations were performed on four different dosage experimental groups and control group at 1h and 3 h after rabbits being poisoned.The echo and the size of kidney were evaluated by two-dimensional ultrasound.The changes of parenchyma were quantitatively analyzed by IBS.Results: No significant changes were found in kidney echo of any groups.With the increasing dosage and prolonging time of poisoning,significant changes were found in volume and length of kidneys comparing with that of control group(P

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The effect of transcription factor SP1 decoy oligodeoxynucleotides on expression of ? Gal in SV-40-PED cells

Yabing HUANG ; Lu WANG ; Lin XIE ; Tao PAN ; Hui GUO ; Rong LI ; Song CHEN ; Shi CHEN

Journal of Medical Postgraduates.2003;0(05):-.

Objective:To investigate the role of transcription factor SP1 decoy oligodeoxynucleotides(ODN) on expression of ? Gal in SV-40-PED cells.Methods:Immortalized porcine aortic endothelial cells of the PED line were cultured and transfected with ?1,3galactosyltransferase(?1,3GT) specific decoy ODN.Cells transfected with mismatch ODN was used as negative controls.Twenty-six hours later the cells were collected.The expression of ? Gal was determined with fluorescence microscope and Western blot.The expression of ?1,3GT mRNA was examined by RT-PCR.Results:Fluorescence microscopy observed the decreased fluorescence of ? Gal after decoy ODN transfection.Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was(48.2?0.9).It is 52.6% of the mock group(P0.05).Conclusion:?1,3GT gene reduce actually occurs following transfection of decoy ODN.Porcine endothelial cells can be the targets of decoy ODN.

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Comparision of the effects between estrogen and dehydroepiandrosterone on improving atherosclerosis of ovariectomized rabbits

Qun HAO

Journal of Medical Postgraduates.2003;0(05):-.

Objective:To compare the effects between estrogen and Prasterone(Dehydroepiandrosterone,DHEA) on improving atherosclerosis of ovariectomized rabbits. Methods: Thirty female New Zealand white rabbits of three months old were randomly divided into five groups: Group A: normal control;Group B: sham operation + high cholesterol feeding;Group C: ovariectomized + high cholesterol feeding;Group D: ovariectomized + high cholesterol feeding+ estradiol benzoate;Group E: ovariectomized + high cholesterol feeding+ DHEA.Twelve weeks later,all rabbits were sacrificed,the serum TC,TG,LDL-C,HDL-C,NO and estradiol levels were measured.The aortas and uterus were harvested for histomorphometrical evaluation with light and scanning electron microscope.The area ratios of atherosclerotic lesion to endothelium,and the thickness and area ratios of endothelium to the medium were determined by computer. Results: ①Following the treatment for 12 weeks,no significant difference was found in serum TC、LDL-C、HDL-C levels among four groups except in group A.②There were significant differences between the serum triglyceride(TC)levels of group D and group C.③The serum No levels of group D and group E were significantly higher than that of groupC.④The serum estradiol levels of group D and group E were significantly higher than that of groupC.⑤The area ratios of atherosclerotic lesion to endothelium and the thickness and area ratios of endothelium to the medium in group D and group E were significantly lower than that of group C.⑥Endometrial hyperplasia of group D was significant,while there was no such Phenomena in group E. Conclusion: Estrogen and DHEA can alleviate the early pathological changes of atherosclerosis of the ovariectomized rabbits,prevent and reduce the formation of atherosclerosis.

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