Objective To investigate the ex pr ession of nm23-H1 protein and proliferating cell nuclear antigen (PCNA) in huma n glioma cells. Methods Expression of nm23-H1 and PCNA in 53 br ain gliomas were detected by immunohistochemistry. Results The immunohistochemistry staini ng of nm23-H1 protein in low-grade astrocytomas (grades Ⅰ and Ⅱ) was significantly higher than that in high-grade astrocytomas (grades Ⅲ and Ⅳ). The immunohistochemistry staining of PCNA in high-grade astrocytomas was s ignificantly higher than that in low-grade astrocytomas. Conclusion The lower expression of nm23-H1 protein and the higher expression of PCNA are correlated with the pathological grade of glioma cells. The expression of nm23-H1 may be used as a hopeful marker for predicting the metastastic potential of gliomas.

Objective To investigate the function of apoptosis in the renal injury by observing change of renal pathology and ultrastructure in the rat with obstructive jaundice(OJ) and if salvia miltiorrhiza(SM) can lighten the renal dysfunction of obstructive jaundice.Methods A total of 100 adult Sprague-Dawley rats were studied.Among them,the 80 models of obstructive jaundice were established by ligating bile duct(BDL),then divided into two groups: the OJ rats administered daily abdomen injections of SM(1.7g per rat) after operation of bile duct ligation(the SM group,n= 40);the OJ rats receiving only the same normal saline(the OJ group,n=40).The other rats with sham operation receiving only normal saline(the control group,n=20).Three groups of rats were sacrificed in groups at postoperation 1,2,3 and 4 week,respectively.Then serum BUN & Cr were tested and renal change of histopathology and ultrastructure were observed;Apoptosis of renal tissue were assayed by TUNEL method.Results With the time of BDL extending,the value of serum BUN & Cr increased;Apoptotic cells increased in renal tissue.After treatment with SM,the injury degree of renal function and histopathologic changes decreased.Conclusion Obstructive jaundice can lead to renal injury.Apoptosis had an important effect on renal function injury in the rat with obstructive jaundice.Salvia miltiorrhiza can ease the degree of renal function injury.

Objective To investigate the relationship of the expression of Bcl-2 and Bax protein with apoptosis of renal tubular epithelia in rats with obstructive jaundice(OJ).Methods A total of 60 adult Sprague-Dawley(SD) rats were divided into two groups: OJ rats by ligating bile duct(OJ group,n= 40) and the sham-operation rats(the control group,n=20).The two groups of rats were sacrificed at postoperative 1,2,3 and 4 week,respectively.Then serum BUN and Cr levels were tested and renal histopathological and ultrastructural changes were observed.Apoptosis of renal tissues was assayed by TUNEL method.The immunohistochemical Elivision~(TM) technique was adopted for detecting the Bcl-2 and Bax protein expression.Results With the time of OJ prolonging,the value of serum BUN and Cr increased;apoptotic cells of renal tubular epithelia increased.There were obvious differences in Bcl-2 and Bax protein expression of between OJ and control groups(P

Objective To explore the radiosensitivity ofβ-elemene on Colo320 cells transplanted tumor in nude mice and its molecular mechanisms.Methods Colo320 cells transplanted tumor model was established by cell suspension inoculation in nude mice.Then the transplanted mice with 0.8-1.0 cm3 tumor were randomly divided into 4 groups:control,β-elemene (40 mg/kg),radiation (4 Gy),andβ-elemene (40 mg/kg)+ radiation (4 Gy) groups,with four to five mice in each.Tumor weight and morphology were observed in each group.In addition,the apoptosis of tumor cells was measured by TUNEL and the expression of Fas gene was detected by immunohistochemistry.Results The nude mice transplanted tumor model was successfully established.Compared with that in control and simple radiation groups,tumor weight was significantly decreased inβ-elemene combined with irradiation group (P <0.05).At the same time,the apoptosis rate and the expression of Fas gene in tumor cells were significantly increased (P <0 .0 5 )inβ-elemene combined with irradiation group compared with control and simple radiation groups. Conclusion β-elemene could enhance the radiosensitivity of Colo320 cells transplanted tumor in nude mice probably by inducing Fas-mediated apoptosis of tumor cells.

Objective To explore the association between X-ray repair cross complementing group 1 (XRCC1)gene rs25487 locus polymorphisms and nonobstructive azoospermia in Hui minority ethic population of Shaanxi Province.Methods We used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)method for genotyping at XRCC1 gene rs25487 locus in 79 patients with nonobstructive azoospermia and 82 healthy male controls in Hui minority ethic population of Shaanxi Province.Then we analyzed the association be-tween XRCC1 gene rs25487 locus and nonobstructive azoospermia.Results Compared with GG genotype,GA, AA and GA + AA genotypes demonstrated a significantly increased risk for nonobstructive azoospermia (OR =2.286,95% CI 1.1 5 1-4.539;OR =2.202,95% CI 0.753-6.439;OR =2.271,95% CI 1.1 71-4.403),respec-tively.Meanwhile,the A allele frequency was significantly higher in azoospermic patients than in controls (OR =1.582,95% CI 1.005-2.492,P =0.047).Conclusion G→A in XRCC1 gene rs25487 locus is correlated with nonobstructive azoospermia in Hui minority ethic population of Shaanxi province.

Aim To study the effect of PA on the proliferation and maturation of cord blood derived DC in vitro. Methods The monocytes were isolated from human umbilical cord blood and cultured with PA, GM-CSF+IL-4+TNF-?(GTI), GTI+PA (GTIP), respectively. On day 7 of cultivation, the CD1a, CD83,HLA-DR and CD34 antigens expression of cells were analyzed by immunohistochemistry. Result The cells with typical morphological properties of DC can be observed with electron microscope among PA, GTI and GTIP groups. The CD1a and CD83 positive rates of PA group increased significantly, reaching 19.63%?3.61% and 14.52%?5.79% respectively, much higher than that of the control. In addition, compared with GIT group, the positive rate in GTIP group has increased remarkably. Conclusion PA does not only promote the proliferation and maturation of cord blood derived DC, but also cooperate with GTI to enhance the production of DC. PA is a useful activator of DC.

Aim To investigate the pharmacokinetic characteristics of silibinin in Chinese healthy volunteers.Methods Nine Chinese male healthy volunteers were divided into receiving orally a single dose of silibinin capsule corresponded 70,140 and 280 mg of silibinin,respectively,in Latin square design study.After administration of silibinin capsule,the plasma concentrations were determined by HPLC with UV detection.The pharmacokinetic parameters were analyzed by Topfit 2.0 program.Results The linearity of this method was found to be from 3.125 to 10 000 μg·L~(-1) with a lower limit of quantitation(LLOQ) of 3.125 μg·L~(-1) for silibinin.The pharmacokinetic parameters were calculated as the follows:at the three different dosages(70,140 and 280 mg),T_(1/2) was 2.44,2.38 and 2.47 h;C_(max) was 1135.6,2841.1 and 3946.9 μg·L~(-1);T_(max) was 1.35,1.26 and 1.39 h;AUC_(0-11 h) was 1287.2,3337.8 and 5398.5 μg·h·L~(-1);AUC_(0-∞)was 1300.7,3377.1 and 5453.9 μg·h·L~(-1);CL/F was 1062.1,824.7 and 943.2 ml·min~(-1);And V_d was 219.9,167.1 and 212.0 L,respectively.Conclusions The developed method is shown to be sensitive,accurate and simple,and can satisfy the requirement of pharmacokinetic study of silibinin in human.The C_(max),AUC_(0-11 h) and AUC_(0-∞) of silibinin in Chinese healthy volunteers(in ranges of 40～120 mg)are fitted with non-linear kinetic model,while there are no significant differences in T_(1/2) at the three different dosages.

Objective To develop a quantitative immunohistochemistry assay for duck hepatitis B virus core antigen (DHB-cAg)in duck liver tissue.Methods By comparison with no repair antigen and repair antigen with high pressure,microwave and trypsin,the best solution of antigen retrieval was determined.By optimizing the parameter of image acquisition and de-ducting blank area,mean density of yellow areas was calculated using Image-Pro Plus 6.0 software.Using the assay devel-oped to determine the level of DHBcAg in liver tissue from duck infected by DHBV,anti-DHBV activity of DHBcMAb-TAT PTD conj ugate was examined.Results SABC method with no repair antigen was selected,which was better than other methods.DHBcAg expression in duck liver tissue could be objectively and accurately quantified by setting Image-Pro Plus 6.0 software parameters and calculating mean density of yellow areas.By comparison with the differences between mean densityat baseline of treatment and end of treatment,it was showed that DHBcMAb-TATPTD conjugate treatment dose-de-pendently reduced the levels of DHBcAg in liver tissue,which show that the assay developed could effectively evaluate the anti-DHBV activity of agent.Conclusion The immunohistochemistry assay developed in this study can objectively and accu-rately evaluate the level of DHBcAg in duck liver tissue.

Objective To investigate the tumor-suppressive activity of polyactin B (PB) and the life-span of tumor bearing mice treated with PB and the anti-tumor mechanisms of PB by blockage of macrophage in vivo.Methods Thirty-two S180 sarcoma-bearing (SC) BALB/c inbred mice were divided into 4 groups, i.e., control group (8 mice), PB group (8), Trypan blue (TrB) group (8) and PB+TrB group (8). Five days after PB injection (ip), the tumor size of the mice was measured, and the ratio of TNA to TTA in HE section of the tumor maximal area was calculated and the pathological features of the tumors were observed. Forty S180 sarcoma-bearing (ip) ICR mice, which were randomly divided into 4 groups as above, were used for observation of the life-span. Results PB could strongly inhibit the growth of S180 sarcoma with the tumors regressing completely in 3 of 8 mice, prolong the life-span of mice bearing S180 sarcoma, increase the infiltration of macrophages and lymphocytes in the periphery and inside of the tumors. In addition, there were more prominent hemorrhagic necrosis, neutrophil infiltration and microthrombi in small vessels around the necrotic areas in the tumors of PB treated group. The tumor-suppressive effect of PB disappeared after macrophage system blockage with TrB. In PB+TrB, TrB, control and PB groups, the size of tumors reduced, and the life-span of mice increased in successive order.Conclusions (1) PB is a strong activator of macrophage. (2) In vivo the tumor-suppressive effect of PB is initially mediated by PB activated macrophage system. Then other immunocompetent cells, e.g., T-cells, NK-cells, etc, activated directly and/or indirectly by PB, may play a very important role in tumor suppression. (3) PB-activated macrophages are more sensitive to TrB blockage. Therefore, using TrB+PB to block macrophage system is more effective than TrB alone. (4) Blockage of macrophage function might influence the life-span of the mice.

Heart failure is the leading cause of death worldwide. Compound Danshen Dripping Pill (CDDP) or CDDP combined with simvastatin has been widely used to treat patients with myocardial infarction and other cardiovascular diseases in China. However, the effect of CDDP on hypercholesterolemia/atherosclerosis-induced heart failure is unknown. We constructed a new model of heart failure induced by hypercholesterolemia/atherosclerosis in apolipoprotein E (ApoE) and LDL receptor (LDLR) dual deficient (ApoE^-/-LDLR^-/-) mice and investigated the effect of CDDP or CDDP plus a low dose of simvastatin on the heart failure. CDDP or CDDP plus a low dose of simvastatin inhibited heart injury by multiple actions including anti-myocardial dysfunction and anti-fibrosis. Mechanistically, both Wnt and lysine-specific demethylase 4A (KDM4A) pathways were significantly activated in mice with heart injury. Conversely, CDDP or CDDP plus a low dose of simvastatin inhibited Wnt pathway by markedly up-regulating expression of Wnt inhibitors. While the anti-inflammation and anti-oxidative stress by CDDP were achieved by inhibiting KDM4A expression and activity. In addition, CDDP attenuated simvastatin-induced myolysis in skeletal muscle. Taken together, our study suggests that CDDP or CDDP plus a low dose of simvastatin can be an effective therapy to reduce hypercholesterolemia/atherosclerosis-induced heart failure.