Objective:To investigate the reversing effect of recombinant mutant human tumor necrosis factor (rmh-TNF) on cisplatin(DDP)-resistant human ovarian cancer cell line SKOV3/DDP in vitro and the related mechanism. Methods: DDP-resistant human ovarian cancer cell line SKOV3/DDP was cultured in vitro. The cytotoxic effect of rmh-TNF to SKOV3/DDP cells was examined by MTT assay and the nontoxic dose of rmh-TNF was identified. The changes of DDP resistance was observed after cells were treated with nontoxic dose of rmh-TNF by MTT assay. The expre-ssion of GST-? protein was examined by flow cytometry at different periods after rmh-TNF intervention; RT-PCR was used to analyze the expression of MDR1gene in SKOV3/DDP cells before and after rmh-TNF treatment. Results:(1)rmh-TNF at 50-122.34 U/ml showed no evident inhibitory effect on the growth of SKOV3/DDP cells (the cell survival rate higher than 90%); and 100 U/ml was chosen for the reversing experiment ( nontoxic dose).(2)IC50 values of SKOV3/DDP cells were (23.29?0.43), (8.97?0.69) and (6.43?0.79) ?g/ml after treatment with DDP for 24, 48 and 72 h, respectively; and the values decreased to (19.50?0.50),(4.34?0.43) and (2.44?0.02)?g/ml after combined treatment with 100 U/ml rmh-TNF, respectively.(3)Expression of GST-? protein and MDR1gene decreased with the prolongation of rmh-TNF treatment. Conclusion: rmh-TNF has reversal effect on the DDP-resistant cell line SKOV3/DDP, and the mechanism may be associated with the down-regulation of GST-? protein and MDR1gene expression.

Objective To explore the relationship between metabolic syndrome and ApoA5 gene polymorphism and serum concentration. Methods 100 patients with metabolic syndrome(MS group) and 100 healthy people(control group) were enrolled in this study. The ApoA5 serum concentration of two groups were measured by using enzyme-linked immunosorbent assay(ELISA), the genotypes of ApoA5-1131T＞C polymorphism were analyzed by polymerase chain reaction-restriction fragment length polymorphism, and fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL) and white blood cell, hemoglobin,platelet, liver and kidney function were measured. Results MS group was compared with control group. In MS group, the ApoA5 serum concentration was significantly lower[(96.68±18.09)ng/ml vs (128.32±23.78)ng/ml,P＜0.01], while the triglycerides levels were obviously higher[(2.35±1.07)mmol/L vs (1.62±1.13)mmol/L,P＜0.01], and ApoA5-1131C allele frequency was higher than that in control group (30% vs 16.5%,P＜0.05). Conclusions The ApoA5 serum concentration in metabolic syndrome was decreased, and ApoA5-1131C was associated with metabolic syndrome.

Objective: To assess therapeutic effect of alprostadil on improving myocardial microcirculatory disturbance and hemorheological disorder in patients with coronary heart disease (CHD).Methods: A total of 164 CHD patients treated in our hospital were selected.According to random number table method, they were randomly and equally divided into routine treatment group and alprostadil group (received alprostadil injection based on routine treatment group), both groups were treated for two weeks.Serum levels of nitric oxide (NO), vascular endothelial growth factor (VEGF), thromboxan β2 (TXβ2) and hemorheological indexes were measured and compared between two groups after treatment;TIMI grade and TIMI myocardial perfusion grading (TMPG) of CAG were compared between two groups after treatment.Results: Compared with routine treatment group after treatment, there were significant rise in serum levels of NO[(64.9±10.3) mmol/L vs.(98.8±13.2) mmol/L]and VEGF[(1.62±0.53) mg/L vs.(3.31±0.68) mg/L], and significant reduction in serum TXβ2 level[(180.4±22.8) pg/ml vs.(78.9±9.6) pg/ml], P<0.05 or<0.01;significant reductions in whole blood high shear viscosity[(5.84±0.72) mPa·s vs.(4.25±0.31) mPa·s], whole blood low shear viscosity[(8.42±0.93) mPa s vs.(5.31±0.68) mPa s], plasma viscosity[(2.73±0.34) mPa s vs.(1.61±0.29) mPa s], fibrinogen level[(4.09±0.55) g/L vs.(3.13±0.55) g/L], erythrocyte aggregation index[(3.85±0.47) vs.(2.24±0.31)]and maximum platelet aggregation rate[(67.4±5.3)% vs.(48.0±3.6)%]in alprostadil group, P<0.05 all.Among those patients undergoing second CAG, compared with routine treatment group, there were significant reductions in percentages of TIMI and TMPG grade II and grade III, and significant rise in percentages of TIMI and TMPG grade 0 and grade I in alprostadil group, P<0.05 or <0.01.Conclusion: Alprostadil can significantly improve myocardial microcirculatory disturbance and hemorheological disorder in patients with coronary heart disease, which is worth extending.

Objective To analyze and investigate dopamine combined with milrinone treatment effect on elderly patients with intractable heart failure and N-terminal B-type natriuretic peptide level and cardiac function.Methods 80 cases of elderly patients with heart failure according to the number table method randomly divided into two groups: control group and experimental group, and control group was given conventional drugs.The patients in experimental group were received dopamine +Milrinone on the basis of control group.Clinical efficacy, N-terminal pro-B-type natriuretic peptide levels ( NT proBNP ) and heart function condition between two groups are compared and analyzed.Results The total effective rate of experimental group (95.00%) was higher than that of control group (70.00%) (P<0.05).NT-proBNP(2013.31 ±295.84)ng/L、LVEDD(61.48 ± 10.11)mm、LVEF(59.69 ±8.44)% in the experimental group was significantly better than the control group(P <0.05).Conclusion Dopamine combined with milrinone in the treatment of elderly patients with intractable heart failure is remarkable, can relieve the level of NT proBNP, and promote the recovery of cardiac function.

Objective To establish the rapid PCR am plification program and system and to verify the technical indexes. Methods PCR m ultiplex and capillary electrophoresis detection of 24 autosom al STR loci and one Y-STR loci using the 6-color fluorescence m arking technology, as w ell as Amelogenin and Y-InDel. Meanw hile, sensitivity, specificity, identity, stability, m ixing and a batch of sam ple tests w ere investigated, and the genotype of various routine sam ples and degraded, exfoliated cell sam ples w ere observed. Results The sensitivity of the system w as 0.062 5 ng. In addition, the genotype could be detected accu-rately only around 65 m in via rapid am plification. The species-specificity w as high and the genotyping of all kinds of dry blood specim ens of filter paper and m ixed, degraded, exfoliated cell sam ples w ere accu-rate. Conclusion The rapid am plification system can significantly im prove the detection rate, and obtain accurate and stable genotyping results, w hich m ay be im portant im plications for the establishm ent of STR database and study on population genetics and forensic identification.

Objective To study the effect of apolipoprotein A5 gene polymorphism on the metabolic syndrome and cardiovascular events.Methods A total of 200 patients with metabolic syndrome include 100 cases of unconsolidated cardiovascular events (MS-1 group) and 100 cases of combined cardiovascular events (MS-2 group); 100 cases of healthy people were used as control group.Apolipoprotein A5 serum concentration was detected by ELISA in each group.PCR-RFLP method was used to determine ApoA5-1131T ＞ C gene polymorphism.The automatic analyzer was used to measure fasting plasma glucose (FPG).Results The MS-1-group ApoA5 Serum concentration was significantly lower than the control group [(96.68 ± 18.09) ng/ml vs (128.32 ±23.78) ng/ml,t =10.59,P ＜0.01] ; the MS 2 groups ApoA5 serum concentrations was significantly lower than the MS-1 group [(87.67 ± 17.09) ng/rnl vs (96.68 ±18.09) ng/ml,t =3.62,P ＜0.01] ; and ApoA5-1131C genotype frequency in MS-2 group and the MS-1 group was significantly higher than the control group (39.4％ ＞ 30％ ＞ 16.5％,P ＜ 0.05).Conclusions ApoA5 serum concentrations in patients with metabolic syndrome and cardiovascular events were significantly reduced,ApoA5-1131 C was related to the metabolic syndrome and cardiovascular events.

Objective:To investigate the effects of pregnancy and lactation nonylphenol (NP) exposure on the balance of Treg/Th17 cells in the brain of offspring mice and the related mechanisms.Methods:Thirty pregnant C57BL/6 mice were randomly divided into three groups: control group (drinking distilled water), and NP-treated groups (drinking 0.2 μg/ml or 2.0 μg/ml NP water solution). ELISA kit was used to analyze the levels of TNF-α, IFN-γ and IL-17, flow cytometry was used to analyze the frequency of Treg and Th17 cells in spleen, quantitative RT-PCR was used to analyze the RORγt, Foxp3 mRNA, Western blot was used to analyze the protein expression of RORγt, Foxp3 and PI3K/Akt/mTOR signal pathway, and immunofluorescence was used to analyze the expression of Iba1 in the brain tissue of offspring mice.Results:Compared with the control group, NP exposure increased the serum levels of IL-17 and TNF-α in male offspring mice ( P<0.05), and decreased the levels of IFN-γ( P<0.05). Flow cytometry analysis showed that the percentage of Th17 cells in the spleen of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly higher than that of the control group, while the percentage of Tregs cells was lower. Compared with the control group, the expression levels of Foxp3 proteins in the brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly lower, accompanied by a dramatic increase in RORγt protein levels ( P<0.05). Similar mRNA expression was also observed in qRT-PCR analysis. The protein expression levels of mTOR (p-mTOR) and its upstream related regulators[PI3K, p-Akt (Ser473), p-Akt (Thr308)] in the brain of male offspring mice increased gradually during the period of exposure to NP( P<0.05). Immunofluorescence analysis showed that compared with the control group, the number of Iba1 positive cells in brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) increased significantly ( P<0.05). Conclusions:Maternal exposure to NP during pregnancy and lactation may affect the development/function of neurons in offspring through neuroimmune axis, and increase the risk of neurodevelopmental disorders in offspring.

Objective:To search the release in vitro of new MU-AN ophthalmic gel consist of ganciclovir instead of aciclovir is whether better than the Old. Methods:Using the content of ganciclovir and acyclovir as the index, taking the second oar method ( in Ch. P 2010), drug release in vitro test was investigated. Results:The character of drug release of new MU-AN ophthalmic gel was e-qual to the old, the rate of drug release was similar, The amount of drug release was the same. Both drugs met the requirements of clin-ical medication. The character corneal permeability of new MU-AN ophthalmic gel was better than the old. Gel matrix had no influ-ences on drug release, drug would be bring treatment effect after the way that it was released quickly then was dissolved in tear. Con-clusion:The drug release characteristics consistent with ophthalmic preparation requirements. The character of drug release of new MU-AN ophthalmic gel consist of ganciclovir instead of aciclovir is equal to the old, the time administer drug and interval time is gener-ally scientific, reasonable and feasible, providing the basis for the pharmacodynamics , toxicology and clinical study in the next step.

Objective:The roles of HMGA2 in maintaining malignant phenotypes of the osteosarcoma U2OS cells was studied to explore the possibilities for it to be developed as a target for gene therapy.Methods:U2OS cells were stably transfected with a DNA based shRNA expression vector which targeted to HMGA2.The expression of HMGA2 mRNA was proved by RT-PCR;Cell growth,migration and apoptosis were determined with CCK8,hoechst33342 staining and Boyden ventricle,respectively.The mRNA levels of Caspase 3 and Caspase 9 were determined by real time quantitative RT-PCR.Results:The transfection with shRNA expression vector significantly decreased HMGA2 mRNA levels of U2OS cells.Cell growth and migration were decreased,but apoptosis and the mRNA levels of Caspase 3 and Caspas 9 were increased following the decrease of HMGA2 mRNA.Conclusion:The abnormal expression of HMGA2 plays an important role in maintaining the malignant phenotypes of U2OS cells.Gene therapy targeted to HMGA2 could be helpful in the treatment of human osteosarcoma.

Objective · To detect the vitamin D2 and D3 levels in the patients with moderate and severe depression. Methods · Eighty-five patientswho met the criteria for major depressive disorder were recruited (53 patients with moderate depression, 32 patients with severe depression). Fifty agematchedhealthy volunteers were recruited as controls. Serum 25 (OH) D2, 25 (OH) D3 and total 25 (OH) D2/D3 levels were detected by using liquidchromatography-tandem mass spectrometry (LC-MS/MS). The risk factors which might influence the severity of depression were screened by Logisticregression analysis. Results · The serum 25 (OH) D2, 25 (OH) D3 and total 25 (OH) D2/D3 levels in the case group was lower than those in the controlgroup (P=0.012, P=0.000, P=0.000). The patients with moderate depression presented significantly lower serum 25 (OH) D3 and total 25 (OH) D2/D3 levelsthan the controls did (P=0.000), although no significant difference in serum 25 (OH) D2 levels was found between these two groups. As well, the serum 25 (OH) D2, 25 (OH) D3 and total 25 (OH) D2/D3 levels in patients with severe depression were significantly lower than those in patients with moderate depression and controls (P<0.05). The body mass index of severe depression group was much higher than that of moderate depression group and control group (P=0.002). Both overweight/obesity and the concentration of vitamin D may be the major influencing factors of depression severity (P=0.034, P=0.011). Conclusion · Vitamin D2 and D3 deficiency in depressive patients, particularly in those patients with severe depression, was shown in the present study. In addition, overweight/obesity as well as the concentration of vitamin D may exert the significant influence on the severity of depression. Vitamin D supplementation and weight control may be needed to be considered in making therapeutic strategies of major depressive disorder.