Abstract:Objective:To observe the influence of lentiviral vector-mediated RNA interference on expression of CTGF(connective tissue growth factor,CTGF)gene in human hepatocellular carcinoma cell line. Methods:The effective RNA interference sequence targeting CTGF gene was screened and determined according to our previous study. The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was desiged,synthesized and cloned into Plk0.1-GFP-SP6 vector to construct a lentiviral vector which expressed CTGF shRNA,subsequently confirmed by PCR and DNA sequencing analysis. 293T cells were co-transfected with the plasmids using liposome transfection methods,and packaged to produce lentiviral particles. The recombinant lentiviruses were transfected into HepG2 cells,and the CTGF mRNA and protein expression were examined by Real-time PCR and Western-blot. The results were compared with those of the non-transfected and blank vector transfected HepG2 cells. Results:PCR analysis and DNA sequencing confirmed that the CTGF shRNA sequences were successfully inserted into the lentiviral vectors. After transfection with CTGF shRNA,the CTGF expression in HepG2 cells was significantly inhibited at both mRNA and protein levels compared with that in non-transfected and empty vector transfected HepG2 cells. Conclusion:Two lentiviral RNAi vectors targeting CTGF gene have been successfully constructed,and can effectively inhibit the expression of CTGF gene in HepG2 cells. It paves a way for CTGF-targeted gene therapy of human hepatocellular carcinoma.

Abstract:Objective:To construct a recombinant prokaryotic vector of GRO--琢 and optimize the condition for expressing GST fusion protein in E.coli BL21(DE3),and discuss the function of GRO--琢-GST fusion protein to promote proliferation of tumor cells. Methods:GRO--琢 gene was amplified by RT-PCR method and inserted into the prokaryotic express vector pGEX-4T-1. The plasmid was transformed into E.coli BL21(DE3) and induced to express fusion protein GRO--琢-GST with IPTG. The fusion protein was detected using SDS-PAGE and Western-blot methods. The fusion protein was purified by GSTrap affinity column,and its function on tumor cells proliferation was detected by CCK-8 method in MCF-7 cells. Results:Recombinant pGEX-4T-1/GRO--琢 vector has been successfully constructed. CCK-8 analysis showed that the fusion protein induced proliferation of MCF-7 cells. And GRO--琢-GST fusion protein with high purity was obtained. Conclusion:The protein has a biological activity to induce proliferation of MCF-7 cells,and the study of GRO--琢-GST protein may facilitate further research on its biological functions.

Abstract:Objective:To explore the expression and significance of Shh and Smo in Sonic Hedgehog(SHH)signaling pathway in hepatocellular carcinoma(HCC). Methods:The expression of Shh and Smo were detected in 10 fresh HCC tissues and 2 HCC cell lines of HepG2 and Huh7 by using RT-PCR,and also observed in 30 HCC tissue slices and 10 normal liver tissue slices using immunohistochemistry method. Results:The mRNA expressions of Shh and Smo were higher in HCC than in adjacent-tumor tissues(P < 0.05). The mRNA expressions of Shh and Smo were higher in Huh7 cell lines than in HepG2,but no significant difference was observed between two cell lines(P > 0.05). Immunohistochemistry results showed in 30 HCC tissue slices,the positive expression rates of Shh and Smo protein were 63.3%(19/30) and 76.7%(23/30),respectively. The expression of Smo protein was significantly correlated with the tumor size(P < 0.05). Shh and Smo proteins did not expressed in normal liver tissue. Conclusion:The expressions of Shh and Smo were elevated in HCC,which indicated that SHH signaling pathway may involve in human HCC carcinogenesis. The study may be useful for treatment of HCC.

Abstract:Objective:To investigate the effect of cellular ATP depletion on actin phosphorylation in rat renal proximal tubule cells(NRK52E). Methods:NRK52E cells were treated with 0.1 μmol/L Antimycin A to established a cellular ATP depletion model. The contents of G-actin(in cytosolic fraction)and F-actin(in cytoskeletal fraction)in cells were evaluated by Western blot. Two-dimensional electrophoresis was used to separate phosphorylated actin from unphosphorylated actin of cells. With immunoprecipitation(IP)and Western blot analysis,residues of actin phosphorylated was explored by using monoclonal anti-phosphoserine,anti-phosphotyrosine and anti-phosphothreonine antibody. Results:The cellular F-actin was increased and G-actin was decreased after ATP depletion. The proportion of phosphorylated actin after ATP depletion was higher than that of control cells. As compared to the control cells,the level of actin phosphorylation on thyrosine residue(s) was significantly increased during cellular ATP depletion,and these changes were paralleled with the severity of cellular ATP depletion. Conclusion:The thyrosine phosphorylation of actin of NRK52E cells may play an important role in regulation of polymerization of actin during ATP depletion.

Abstract:Objective:To detect the effect of a new triterpene saponins extracted from Clematis ganpiniana on growth and apoptosis of human gastric carcinoma cell line SGC-7901,and investigate the effect of Clematis saponins on the activity of nuclear transcription factor NF-κB of SGC-7901 cells,and preliminarily explore its possible mechanism. Methods:After SGC-7901 cells were treated with Clematis saponins at different concentrations for 48h,MTT assay was used to detect the growth inhibition of cells. Negative control and blank control were set. Apoptosis rate of SGC-7901 cells treated with Clematis saponins and 5-fluorouracil(5-FU)was determined by Annexin V/PI double staining of flow cytometry(FCM). Western blot was used to test the expression of NF-κB p65 in nuclei. The intranuclear activity of NF-κB was confirmed by ELISA assay. Results:The inhibition rate on SGC-7901 cells in each Clematis saponins treatment group was significantly higher than that in control group(P < 0.05),and the inhibition rates increased with the concentration increase of Clematis saponins. The Clematis saponins could obviously promote cell apoptosis induced by 5-FU. Western blot and ELISA results confirmed that the Clematis saponins could significantly inhibit the expression and activity of NF-κB in nuclei. And the inhibition pattern showed in a dose-dependent manner. Conclusion:Clematis saponins can significantly inhibit cell proliferation and induce cell apoptosis of gastric carcinoma cell line SGC-7901,which might be partly related to the inhibition of the NF-κB expression.

Abstract:Objective:To study the expression and function of B7H3 in human myeloid leukemic cell lines,U937 and K562. Methods:The protein and mRNA expression levels of B7H3 were detected by FACS and RT-PCR methods,respectively. The effect of B7H3 in human leukemic cell lines was studied by B7H3 mAb inhibiton assay. Cell proliferation was determined by cell counting. Results:B7H3 expressed in both U937 and K562 cell lines. B7H3 mAb could inhibit cell proliferation of those two cell lines. Conclusion:B7H3 is constitutive expression in human myeloid leukemia cell lines,U937 and K562. B7H3 mAb can conjugate with B7H3 expressed in cell surface,which plays a critical role in the survival and growth of leukemia cells.

Abstract:Objective:To study the gene expression of sodium/iodide symporter(NIS) in both differentiated thyroid carcinoma(DTC)and correlated lymph nodes metastasis,and explore the relationship between NIS expression and DTC,and between NIS expression and the significance of radioactive iodine treatment for DTC. Methods:The expression of NIS in tissue specimens was evaluated by RT-PCR technique and immunohistochemistry. Results:The RT-PCR result showed that the expression of NIS mRNA didn’t have significant difference between tumor-adjacent tissues and DTC tissues. The NIS mRNA expression in metastatic lymph nodes were lower compared with tumor and tumor-adjacent tissues. The immunohistochemistry result showed that NIS positive ratio in DTC and correlated lymph nodes tissue didn’t have significant difference compared with that in normal thyroid,respectively. Conclusion:The NIS could be normally expressed,but NIS protein was mostly expressed in the cytoplasm rather than in the functional membrane,which has guiding significance for the radioiodine treatment of DTC.

Abstract:Objective:To explore the expression of vascular endothelial growth factor C(VEGF-C) and lymphatic micro-vessel density in their clinical significance of melanoma tissues. Methods:The expression of VEGF-C and D2-40 labeled lymphatic micro-vessel density in melanoma were determined by immunohistochemistry. Both the correlation between VEGF-C and lymphatic micro-vessel density and the relation of their expression with clinical and pathological features of melanoma were analyzed. Then their effect on metastasis-free survival rate in melanoma were separately evaluated by survival analysis. Patients with nevi were used as control. Results:The lymphatic micro-vessel density in melanoma(14.30±4.91)was more than that in the nevi(7.64±3.08)(P < 0.01). The lymphatic micro-vessel density were higher in ulcer group than in non-ulcer group(P=0.005).There was no significant correlation between VEGF-C expression and clinicopathological characteristics(P > 0.05). The intensity of VEGF-C expression has no effect on lymphatic micro-vessel density and metastasis-free survival rate(P > 0.05). The metastasis-free survival rate in patients with melanoma,whose lymphatic micro-vessel density is more than or equal to 14.7,was significantly shorter than those less than 14.7. Conclusion:Lymphatic vessels were found within tumor. The lymphatic micro-vessel density is positively correlated with ulcer,and can predict lymph node metastasis of melanoma.

Abstract:Objective:To investigate the clinical significance and the expressions of the lung resistance protein(LRP),glutathione-S-transferase π(GST-π),excision repair cross-completion group 1(ERCC1)protein and Survivin protein in non-small cell lung cancer(NSCLC). Methods:LRP,GST-π,ERCC1 and Survivin protein in 64 specimens from NSCLC patients were detected using Envision immunohistochemical method,and the relationship was analyzed between the different protein expressions and the pathological classification,histological type,lymph node metastasis,clinical stage and the efficacy of platinum-based chemotherapy of NSCLC. Results:The positive rates of LRP,GST-π,ERCC1 protein and Survivin protein were 60.9%,56.3%,60.9% and 54.7% respectively. High expression of LRP was associated with poorly differentiated type(85.0%). The positive rate of GST-π decreased with the degradation of differentiation degree of NSCLC. The significant high positive rate of ERCC1 protein was observed in squamous cell carcinoma(80.0%). The positive rate of Survivin protein was significantly high in adenocarcinoma cell carcinoma(81.5%). There were significant differences in those proteins expressions among different carcinoma types. There was a positive correlation between the expression of LRP and the expression of GST-π(R=0.262,P=0.036). High expressions of ERCC1 protein and LRP were two independent factors related to platinum resistance by multivariate regression analysis. In ERCC1 negative NSCLC,the LRP positive patients’ remission rate of platinum-based chemotherapy was lower than the LRP negative patients’(P < 0.05). Conclusion:The expression levels of these proteins varied in different types of NSCLC. There was coordinate expression of LRP and GST-π. The combined detection of ERCCI and LRP proteins could be used to predict the sensitivity to platinum-based chemotherapy.

Abstract:Objective:To explore the expression level of CD58/CD2 and its relationship with the clinical pathological characteristics. Methods:Among 40 patients with uterine cervix cancer and 30 CIN patients,immunohistochemistry method was used to detect the expression of CD58,and flow cytometry was used to detect the CD2 expression in peripheral blood cells,and 30 cases of hysteromyoma were collected as normal controls. Results:The expressions of CD2 and CD58 in patients with uterine cervix cancer were significantly lower than those in CIN patients and normal controls,respectively(P < 0.05). The expression levels of CD58/CD2 were different in uterine cervix cancer patients with different pathological grades,tumor size and invasion depth(P < 0.05). But there were no significant differences in uterine cervix cancer patients with different FIGO stage,age and menopause situation respectively(P > 0.05). Conclusion:The expressions of CD58/CD2 were correlated with the development of uterine cervix cancer,and they could be used as the index for evaluating treatment efficacy and prognosis,and may be the new targes for therapy.

Abstract:Objective:To study the metabolic support function of transplanted hepatocytes labled by green fluorescent protein in the animal model of radiation-induced liver damage(RILD) in vivo,and evaluate the therapeutic effects of transplanted hepatocytes in RILD. Methods:The models were constructed by exposing rats liver to 60Co irradiation,the dose being 40 Gy,and the dose ratio 100 cG/min. Four days later,the rats underwent partial hepatectomy,and were divided into two groups randomly. In the experimental group,5.0×106 hepatocytes transfected by Ad-EGFP were injected into the spleens while the control underwent physiologic saline injection in the same places. Blood samples were obtained at an interval of 5 weeks to measure the serum albumin level of the two groups,and histopathological changes were evaluated by light and electron microscopic examination. The survival rates of the two group were observed dynamically. Results:One week after hepatocytes transplantation(HT),serum albumin level of the experimental group ascended obviously,having significant differences with that of the control. It still ascended progressively 5 weeks later,showing significant differences with that of the control,as well as itself 5 weeks earlier. The differences of mortality rates between the two groups in 1 week and in 5 weeks after HT both show statistical significance. Histological examination suggested that radiative damage were ameliorated in the experimental group compared with that in the control. Conclusion:After transplanted into the spleens of the RILD model,congeneic rat hepatocytes can provide fairly good metabolic support and raise the short-term survival rate in the rat model.

Abstract:Objective:To investigate the effects of exogenous vascular endothelial growth factor(VEGF)on hepatic ischemia/reperfusion injury in rats. Methods:Thirty normal male Sprague-Dawley rats were randomly divided into three groups:group A that was subjected to ischemia/reperfusion injury with intraperitoneal administration of VEGF(50 μg/300 g)30 min before ischemia;group B with the same injury followed by saline administration in the same manner,and group C with only anesthetization leparotomy without ischemia. Animals were killed at 6 hours after reperfusion. Routine assays were performed for testing the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),lactate dehydrogenase (LDH),the activity of hepatic myeloperoxidase(MPO)and induced nitric oxide synthase (iNOS).The pathological changes in the liver were evaluated in hematoxylin and eosin(HE)stained sections. Results:The levels of serum ALT,AST,LDH and the activity of iNOS,MPO in hepatic tissues increased significantly after hepatic ischemia/reperfusion injury. But these effects were offset by administration of VEGF(P < 0.01). In group B,widespread pathological changes and cell apoptosis were observed in the hepatic tissue following hepatic ischemia/reperfusion injury,while similar changes were scarcely visible in groupe A due to the protective effect of intraperitoneal administration of VEGF 30 min before ischemia. Conclusion:The exogenous VEGF can depress the activity of iNOS and alleviate the liver function during the ischemia/reperfusion period,and protect the liver from ischemia and reperfusion injury.

Abstract:Objective:To construct a simple,safe,high successful rate model of renal acute ischemia-reperfusion injury in Wistar rats. Methods:Renal artery of Wistar rats was bilaterally clamped with a micro-artery clamp for 50 minutes to make the animal model of renal acute ischemia-reperfusion injury. Then these kidneys were removed at the 3rd,5th,7th day after injury. Immunohistochemistry staining was used to show the distribution and expression of Kim-1 antigen,and the changes of cytomorphology were observed by HE staining. Results:The successful rate of model construction reached to 85% in the experiment. The expression of Kim-1 antigen increaced. Meanwhile,changes of cytomorphology took place. Conclusion:Stable model of renal acute ischemia-reperfusion injury of Wistar rats can be constructed by applying micro-artery clamp to incarcerate bilateral renal arteries,and its successful rate is very high.

Abstract:Objective:To explore the protective effect of thyroid hormone supplement on the hepatocytes in a rat model of acute hepatic failure. Methods:Thirty-six SD rats were randomly distributed in three groups. In group-1(control group),the rats received only placebo. In group-2(AHF group),the rats received intraperitoneal injection of D-galactosamine and saline. In group-3(treatment group),the rats received intraperitoneal injection of triiodothyronine(T3)at a dose of 15 mg/kg in addition to D-galactosamine. Blood and liver samples were obtained in all animals for the determination of thyroid and hepatic function and assessment of hepatic injuries. Results:T3 was subnormal in rat model of acute hepatic failure. There were less hepatocyte injuries and lower mortality in the treatment group as compared to the AHF group(P < 0.05). Conclusion:A rat model of acute hepatic failure has euthyroid sick syndrome. Thyroid hormone supplement may improve the prognosis and have significant hepatocyte protection in this animal model of acute hepatic failure.

Abstract:Objective:To observe the effects of islet-liking cell clusters inducing from pancreatic duct stem cells in treatment of diabetic rats by transplantation. Methods:After intraductal collagenase digestion,purified by Ficoll density gradient centrifugation,fresh islets and pancreatic duct stem cells were obtained. Then the cells were cultured and expanded in vitro. The 2nd to 6th generation of duct cells were induced in four stages to differentiate into islet-like cell clusters. Islet-like cell clusters were detected by immunofluorescence staining before transplantation. The rat diabetic model was constructed by intraperitoneal injection with streptozotocin(STZ). Eighteen diabetes rats were randomly divided into three groups:diabetic control group,islet transplantation group,islet-liking cell clusters transplantation group. The implants were transplanted into the subcapsular in the left kidneys. The levels of non-fasting blood glucose of rats in 3 groups were measured every other day. Results:The pancreatic duct cells isolated and purified had the characteristic of stem cells by immunofluorescence staining. The results of immunofluorescence staining revealed that insulining and glucagon stain of the islet-like cell clusters were positive. After transplantation,the level of blood glucose of rats in the control group was high,keeping in the range of diabetes. The blood glucose level of rats in the islet transplantation group decreased after transplantation,which was in the normal range for two weeks. The blood glucose level of rats implanted with islet-like cell clusters increased in the first two days after transplantation and decreased after then,and was higher than the normal range during two weeks. Conclusion:Islet-like cell clusters generated in vitro from pancreatic duct cells has some effects on reversing the diabetes in diabetic rats,although the blood glucose control effect is not satisfying.

Abstract:Objective:To investigate the different model construction methods in diabetic nephropathy(DN) SD rats induced by streptozotocin(STZ) and establish the optimal plan. Methods:Thirty SD rats were randomly divided into 3 groups with 10 rats in each group. A group received intraperitoneal STZ after unilateral nephritic excision;B group received intraperitoneal STZ after unilateral nephritic ligation;C group(control group) received buffer solution only. The changes of body weight(BW),glucose(Glu),serum creatinine (Scr),cholesterol(Chol),triglyceride (TG)and 24-urine protein(24Upro),urine crea(Ucr)were detected every 2 weeks and creatinine clearance rate(Ccr) was calculated. After 11 weeks,the rats were sacrificed,and the kidneys were weighed and taken for pathological examination. Results:There was no significant difference in the model construction periods between the groups A and B. The survival rate of the group B is higher than that of the group A. Conclusion:STZ(55 mg/kg) intraperitoneal injection after unilateral nephritic ligation is the optimal method for inducing rat diabetic nephropathy.

Abstract:Objective:To explore effect of combination therapy with vitamin A(Vit A) and insulin on blood glucose in dibetic rats. Methods:Mild diabetes mellitus was induced in male Wistar rats by a single intraperitoneal injection of streptozotocin(STZ) at a dose of 40 mg/kg. Diabetic rats were assigned randomly to the model group,insulin treatment group,Vit A treatment group,Vit A combined with insulin treatment group,and normal rats were used as the control group(n=10 in each group). Insulin treatment was accomplished by subcutaneous injection twice daily for 3 days at a dose of 20 U/(kg·d). Vit A treatment was accomplished by gastric perfusion twice daily for 3 days at a dose of 80 000 U/(kg·d). Serum retinol in various groups was measured by high performance liquid chromatography before and during the interference. Fasting blood glucose and intraperitoneal glucose tolerance test in 5 groups were analyzed at different times following the interference. The pancreas tissue were obtained from 5 groups of rats,and stained immunohistochemically for β cells mass measurement using an image analysis system. Islet isolated and purified from normal rats were cultured with or without retinoic acid. The proliferation of islet cells treated was measured by CCK-8 method. Results:At 2 day,4 day and 6 day following the interference,fasting blood glucose in insulin treatment group and Vit A treatment group had no significant difference compared with that in diabetic model group. And the fasting blood glucose in Vit A combined with insulin treatment group were (6.68±1.82)mmol/L,(7.14±1.27)mmol/L,(6.82±1.36)mmol/L at 2,4 and 6 day of treatment respectively,which were significantly different from those in diabetic model group(P < 0.05). The area under the curve of glucose tolerance in Vit A combined with insulin treatment group was(19.96±4.76)mmol/L,which was significantly different from (42.26±4.38)mmol/L in diabetic model group (P < 0.05). Islet β cells mass in insulin treatment group and Vit A treatment group were(1.78±0.31)mg and(1.67±0.85)mg respectively,which were no significance compared with(1.82±0.25)mg in diabetic model group,and the index in Vit A combined with insulin treatment group was (3.25±0.73)mg,which was significantly higher than that of diabetic model group(P < 0.05). In vitro islet culture showed that retinoic acid did not affect islet cell proliferation. Conclusion:Combination therapy with vitamin A and insulin can normalize fasting blood glucose and glucose tolerance in STZ diabetic rats through increasing Β cell mass.

Abstract:Objective:To explore the effect of Rho-kinase signal pathway in renal interstitial fibrosis of diabetic nephropathy(DN)and the mechanism of fluvastatin in the prevention of renal interstitial fibrosis of DN. Methods:Human renal proximal tubular epithelial cells(HK-2 cells)were cultured in vitro. Rho-kinase activity was expressed as phosphorylation of myosin-phosphatase target-1(p-MYPT-1). The level of p-MYPT-1 and fibronectin(FN) stimulated by high glucose was determined by Western blot at the time of 0 h,6 h,12 h and 24 h after treatment. Same markers were detected when HK-2 cells cultured with high glucose treated with different concentrations(10-7,10-6 and 10-5 mol/L)of fluvastatin for 12 h and treated by lysophosphatidic acid(LPA) further. Results:High glucose enhance the expression of p-MYPT-1 and FN in culured HK-2 cells at the time of 6 h,12 h and 24 h,compared with the time of 0 h. The increase of FN expression stimulated by high glucose was time-dependent and the increased level of p-MYPT-1 reached the peak at 12 h. Fluvastatin decreased the level of p-MYPT-1 and FN induced by high glucose in a dose-dependent manner. The inhibitory effect of fluvastatin on up-regulation of p-MYPT-1 and FN stimulated by high glucose can be reversed by LPA. Conclusion:Rho-kinase may be one of the initiation signals of renal interstitial fibrosis of DN. Fluvastatin can prevent the development of renal interstitial fibrosis of DN by inhibiting Rho-kinase signaling pathway.

Abstract:Objective:To investigate insulin on the viability of ischemic random dorsal flap in the SD rats,and discuss the correlation between the protection of the ischemic-reperfusion injury and the enhancement of viability on the random flap of rats . Methods:Rat model with caudally based random dorsal flap measuring 6.0 cm×1.5 cm was constructed,which were divided into 2 groups:treated group(Group A,n=16)with single-dose insulin 0.1 U(1 ml)injection under the flap afer operation and control group(Group B,n=16)with saline. Before injection and 1 h,2 h,3 h after injection,levels of blood glucose were measured. At 24 h postoperatively,the amount of superoxide dismutase(SOD),glutathione(GSH)and malondialdehyde(MDA)in the flap tissue samples were measured. The flap viability and microvascular expression were measured at 7th day after operation. Results:There was not significant decrease in blood glucose in treated group compared with control group(P > 0.05). At 24 h postoperatively,there was significant decrease in MDA in treated group compared with control group,and significant increase in SOD and GSH(P < 0.05);at 7th day after operation,the viability in treated group was significantly higher compared with that in the control group[(73.963±8.649)% vs(56.175±4.250)%,P < 0.01]. Conclusion:Topical use of insulin markedly improved the viability of random dorsal flap of SD rats,which may be involved in the reduction of ischemia-reperfusion injury.

Abstract:Objective:To compare the features of decidual monocyte-macrophages separated from placental tissues with different gestational ages and explore the role of these immunocytes in immunological tolerance to fetal antigen and the activation of parturition. Methods:We separated decidual cells from healthy decidua samples mechanically,which were obtained after selective abortions at 16 to 25 weeks,or after term deliveries(spontaneous term vaginal deliveries).After purified by MACS,macrophages were analyzed by flow cytometry for molecular makers on cellular surface and cytokines secretion. Their abilities of inducing proliferation were also identified by CCK-8 incorporation assay. Besides,cytokines levels of the cultured allogenic T cells mixed with macrophages were detected. Results:CD14+ macrophages with a purity of 93.32% were obtained. Decidual macrophages origined from normal term-placental tissues expressed low levels of CD80,CD86 and HLA-DR,with no expression of CD83,and secreted low levels of IL-10 and TGF-β. They slightly stimulated allogenic T cells to proliferate and produce low IL-10 and high TGF-β. In contrast,we found higher levels of CD80,CD86,HLA-DR and CD83 expressed on macrophages from mid-pregnancy placenta with higher levels of IL-10 and TGF-β. They slightly stimulated allogenic T cells to proliferate and produce higher IL-10. Conclusion:Monocyte-macrophages separated from placental tissues with different gestational ages are different in biological characteristics. Our results suggests IL-10 may be related to the function of decidual macrophages at mid-pregnancy.

Abstract:Objective:To evaluate the association between the mismatch repair gene hMLH1-93G>A promoter polymorphism and risk of gastric cancer in a population of Jiangsu province in China. Methods:Total of 554 patients with gastric cancer cases and 582 cancer-free controls frequency-matched by age(±5)and sex were recruited in the study. The genotypes of the hMLH1-93G>A polymorphism were detected by TaqMan MGB probe method. We further assessed its association with risk of gastric cancer and interaction with tumor clinic pathological characteristics. Results:There was no significant association of the frequencies of GA or AA among the case and control groups,for case group:GA 262(47.3%),AA 187(33.8%); for control group:GA 269(46.2%),AA 190(32.7%);P=0.636 and 0.398,respectively. The variant genotypes(GA+AA) did not significantly increase the risk of gastric cancer,compared with the GG genotype[adjusted odds ratio(OR)=1.11,95% confidence interval(CI)=0.82~1.51,P=0.487]. No significant association was observed between the variant genotypes of the hMLH1-93G>A polymorphism and the clinic pathological characteristics in gastric cancer. Conclusion:There was no significant association between the hMLH1-93G>A polymorphism and gastric cancer susceptibility.

Abstract:Objective:To establish high throughput mitochondrial DNA(mtDNA)SNPs detection system and study the genetic polymorphism of the Guangdong Han population by SNaPshot technology. Methods:In the present study,we selected 21 SNPs from mitochondrial genome,designed primers,performed PCR,co-amplified single base extension(SBE) and also combined with capillary electrophoresis. Results:Multiple detection system were successfully established. The genetic polymorphism of Guangdong Han population was investigated,and 90 haplotypes were detected in the studied population,and the haplotype diversity(HD) was 97.1%. Conclusion:The results show that the 21-plex mtDNA SNPs multiplex detection system has a pretty good application prospect in forensic medicine.

Abstract:Objective:To examine the prevalence of Ureaplasma urealyticum and Mycoplasma genitalium in male HIV/AIDS patients in Jiangsu Province,in order to provide evidence for future researches on the role of these two mycoplasmas in AIDS. Methods:The organisms were detected from first-void urine specimens in males. Ureaplasma urealyticum was detected by culture method,whereas nested-PCR was used to detect Mycoplasma genitalium. Results:The prevalences of Ureaplasma urealyticum and Mycoplasma genitalium in male HIV/AIDS patients in Jiangsu Province were 51.71% and 28.97%,respectively;the rates of different areas were various. The association of Mycoplasma genitalium infection with the CD4+ cell counts in HIV/AIDS patients was significant. Conclusion:The prevalences of Ureaplasma urealyticum and Mycoplasma genitalium in male HIV/AIDS patients in Jiangsu Province were both high,and Mycoplasma genitalium infection may connect with people’s immune status.

Abstract:Objective:To evaluate the efficacy and safety of moxifloxacin treatment on refractory,recurrent and multi-drug resistant tuberculosis. Methods:The randomized controlled trials of moxifloxacin which is used to treat the refractory,recurrent and multi-drug resistant tuberculosis were recruited. Data were analyzed using RevMan 4.2 provided by the Cochrane Center. Results:Nine RCTs were included. Meta analysis showed that compared with the control group,moxifloxacin was more effective in focus absorption[OR = 1.68,95% CI(1.14~2.49)]and sputum negative conversion[OR=2.47,95% CI(1.61~3.79)]. Adverse effect such as stomach discomfort,nausea,acid reflux,loss of appetite and other gastrointestinal reactions and mild liver dysfunction appeared,and recovered by symptomatic treatment. Conclusion:The current limited evidence suggests that the moxifloxacin treatment on refractory,recurrent and multi-drug resistant tuberculosis is useful for focus absorption and the sputum negative conversion.

Abstract:Objective:To assess the therapeutic effects and adverse effects of mycophenolate mofetil(MMF)and cyclophosphamide(CTX)in the treatment of refractory nephrotic syndrome(RNS) in the systematic review of randomized controlled trials. Methods:We searched PubMed(1966~2009),EMBASE(1984~2009),CBM(1979~2009),CNKI(1994~2009),VIP(1984~2009),CMCC(1994~2009)and Wanfang database(1982~2009)with key words of “mycophenolate mofetil”,“cyclophosphamide” and “refractory nephrotic syndrome” to identify randomized controlled trials of mycophenolate mofetil versus cyclophosphamide for refractory nephrotic syndrome patients. The quality of literature was evaluated,and Meta-analysis was conducted using RevMan 4.2.9 software. Results:Eight RCTs were identified. There was no evidence showing a publication bias except hepatic lesion’s study after Begg’s test analysis. Compared with CTX groups,MMF increased the clinical remission rate,including complete remission rate and partial remission rate(P < 0.05). It also significantly reduced the adverse effects(P < 0.05). Conclusion:The therapy of MMF plus steroids were more effective than that of CTX plus steroids in clinical remission of RNS,and had fewer adverse effects.

Abstract:Objective:To make an analysis and intervention study on the risk factors to the young and middle-aged population with abnormal blood pressure using the advanced health management method. Methods:Total 105 young and middle-aged patients with prehypertension or hypertension received one-year-intervention health management. Then the changes in the intervention factors,such as lifestyle,blood pressure and blood lipids,were analyzed. Results:①After one year intervention,the changes in smoking,alcohol consumption,diet,and exercising status have statistical significances. ②After a whole course of health management,the proportion of hypertension after six months decreased from 52.38% to 23.81%,and further to 5.71% after one year. This intervention is effective(P < 0.000 1). ③The level of blood pressure changed significantly in patients of the simple lifestyle changing group and the receiving antihypertensive treatment group,and also in the patients with prehypertension and hypertension after one year later(P <0.05). ④Besides high-density lipoprotein cholesterol,the risk factors including lipids,glucose and obesity decreased significantly after intervention (P < 0.05). Conclusion:Whole range health management and intervention can change young and middle-aged patients’ lifestyle and control the blood lipids,blood-glucose,obesity and other risk factors,and effectively control blood pressure.

Abstract:Objective:To investigate the clinical effect on resection of stage Ⅲ and Ⅳ gastric cancer after neoadjuvant chemotherapy with the combination of docetaxel,oxaliplatin fluorouracil and leucovorin. Methods:From January,2005 to August,2008,40 patients with stage Ⅲ and Ⅳ gastric cancer received the neoadjuvant chemotherapy 4~6 weeks before operation. Neoadjuvant chemotherapy regimen consisted of 60 mg/m2 docetaxel(Day 1),75 mg/m2 oxaliplatin(Day 5),500 mg/m2 fluorouracil(Day 1 to 5)and 200 mg/m2 leucovorin(Day 1 to 5)administered every 4 weeks for 2~3 cycles before operation. Results:Objective response rates were 70%;Objective pathological response rates were 85%;Total resection rates were 85%;Curative resection rates were 62.5%;TTP was 6.4 months;MST was 20.6 months. Major side effects were myelosuppression,nausea and vomiting. No serious complications were observed. Conclusion:The overall curative resection rates and the compliance were increased in patients with the stage Ⅲ and Ⅳ gastric cancer after the neoadjuvant chemotherapy with the combination of docetaxel,oxaliplatin fluorouracil and leucovorin.

Abstract:Objective:To study the application of 99mTc-MDP bone scintigraphy in multiple myeloma(MM) bone disease. Methods:99mTc-MDP bone scintigraphy and skeletal radiography of 83 newly diagnosed MM patients were analyzed. Results:The positive rate of bone scintigraphy was 88.0%(73/83 cases),and X-ray had a positive rate of 73.3%(55/75 cases),lower than bone scintigraphy. But the positive rate of patients who had multi-site X-ray examination including chest,lumbar and pelvic,had no significant difference(P=0.726)from bone scintigraphy. The positive rate of 99mTc-MDP bone scintigraphy for rib lesions was higher than that of X-ray(P < 0.01). While the positive rate of X- ray for the skull,pelvic lesions was higher than that of bone scintigraphy(P < 0.01). Conclusion:99mTc-MDP bone scintigraphy and multi-site X-ray examination have the similar sensitivity. But scintigraphic images were more sensitive to rib lesions than radiography,while the X-ray was more sensitive in the detection of skull,pelvic lesions. Therefore,the combination of bone scintigraphy and X-ray has obvious benefit to diagnose multiple myeloma. When these two methods do not match each other,CT/MRI can be used to improve the detection rate of MM bone disease.