Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adjuvants, Immunologic ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; HIV-1 ; Humans ; Phytotherapy ; Toll-Like Receptors ; immunology ; physiology

Background Study confirmed that the active microglia may injure retinal ganglion cells (RGCs) in retinal ischemia reperfusion injury (IRI), and increased cx3cr1 expression is an important factor in microglial activation,and thus blocking the expression of cx3cr1 can inhibit microglial activation, which may be useful in neuronal protection.Objective This study was to analyze the protective effects of cx3cr1 antibody on retinal neuron in rat eyes with IRI.Methods Ninety SD rats were divided into 4 groups according to random number table.IRI models were established by perfusing normal saline solution into the anterior chamber.The cx3cr1 antibody of 1 μl (0.2 μg/μl) was intravitreally injected in the right eyes in the normal rats or model rats as the only cx3cr1 antibody injected group and the model cx3cr1 antibody injected group,respectively,and no any drug was injected in the rats of the normal control group and model control group.Retinal sections were prepared 48 hours after modeling, and apoptosis of retinal neutron was observed under the transmission electron microscope;the morphology of retinas was exmined and the number of survival RGCs was calculated by histopathologic method.The expression of CD68 in activated retinal microglial cells was detected by immunochemistry, and the relative expression levels of cx3cr1 mRNA,tumor necrosis factor-α (TNF-ct) mRNA and interleukin-1 β (IL-1 β) mRNA in the retinas were assayed by real time quantitative PCR.Results The cell nucleus of RGCs showed the round and ellipse in shape and there were abundant organelles in the cells.The mophology of photoreceptors was normal with abundant mitochondrions.Irregular cell shape, disrupture of outer segment membranous disc, proliferative microglial cells in RGC layer were seen in the model group.However,these findings were mild in the model cx3crl antibody group.The mean number of survival RGCs was (38.100 ± 3.929), (37.200 ± 5.266), (26.700 ± 2.584) and (31.700 ± 2.946)/field in the normal control group,only cx3cr1 antibody injected group, model control group and model cx3cr1 antibody injected group,showing significant differences between the model group and the normal control group, only cx3cr1 antibody injected group or model cx3cr1 antibody injected group (t =7.492,6.125,-4.607, all at P＜0.01).The expression levels (absorbance) of CD68 in rat retinas were significantly higher in the model group than those in the normal control group, only cx3cr1 antibody injected group and model cx3cr1 antibody injected group (t =-3.397 ,P =0.008;t =-6.207 ,P =0.000;t =3.494, P =0.007).The relative expression levels of cx3cr1 mRNA, TNF-α mRNA and IL-1 β mRNA in rat retinas were raised in the model group compared with the only cx3er1 antibody injected group and model cx3cr1 antibody injected group (all at P＜0.01).No significant differences were observed in these indicators between the normal control group and the only cx3crl antibody injected group (all at P＜0.05).Conclusions Intravitreal injection of cx3cr1 antibody to neutralize cx3cr1 levels in retinas can effectively inhibit the activation of retinal microglia,decrease the release of inflammatory factors, reduce the apoptosis of RGCs and thereby protect the retinal neutrons against IRI in SD rats.Intravitreal injection of cx3cr1 is safe and feasible.

Objective To investigate the effect of macrophage-conditioned medium on the growth of mouse myoblasts. Methods After a 3-day intraperitoneal injection with a mixture of muscle homogenate and starch,macrophages were obtained from mouse peritoneal cavity and cultured for 48 hours in serum-free DMEM/F12.Then the cultured medium was harvested.Myoblasts were obtained from newborn mice skeletal muscle and grown in DMEM/F12 supplied with 10% FBS for 36-48 hours,and then exposed to 0.5% FBS medium with or without macrophage-conditioned medium.Cells were harvested at 72 hours later,their growth was observed with Wright staining,LDH cytochemistry and immunocytochemistry. Results Macrophage-conditioned medium can maintain the normal morphology and the activity of myoblasts grown in low concentration of serum and promote their proliferation.There was a significant difference between average absorbance and integral absorbance of LDH positive cells of the two groups(P

Objective To investigate the histochemical changes of 3?-hydroxysteroid dyhydrogenase(3?-HSD), sucinic dehydrogenase(SDH) and lipid on rat adrenal cortex in the carcinogenesis-resistance with selentum. Methods Wistar rats stomach carcinogenesis (formation of aneuploid cells in the glandular stomach mucosa) was induced by MNNG(N-methy1-N-nitroso-guanidine,20 mg/kg)gavage. The changes of the adrenal cortex were studied by histochemistry and image analysis during preventing glandular stomach cancer with selenium(0.1 mg/kg and 2.0 mg/kg) Results 1.Dietary Se inhibited the aneuploid cell formation,which induced by MNNG gavage in Wistar rat stomach; 2.The histochemical reactions of 3?-HSD and SDH were both significantly stronger in experimental group than that in normal control group in the course of the rat stomach carcinogenesis(P

AIM: To investigate the effect of high glucose toxicity on JNK pathway and cell function of INS-1 cells.METHODS: Cultured INS-1 cells with or without IGF-1 exposure, were treated with glucose at 3 concentrations (5.6 mmol/L, 11.2 mmol/L and 33.3mmol/L), respectively. MTT was used to measure the cell viability. Apoptosis was determined by immuno-fluorescence and flow-cytometry analysis. The serine 270 phosphorylation of IRS and phosphorylation of JNK in INS-1 cells were detected in the presence or absence of SP600125 treatment.RESULTS: The cell viability decreased and apoptosis increased with elevated glucose concentrations. The percentage of apoptosis cells was 11.3% in 5.6 G group, 12.7% in 11.2 G group and 28.2% in 33.3 G group. There was remarkable increase in apoptosis in 33.3 G group with a 2.49-fold increase to the cells in the basal 5.6 mmol/L glucose. High glucose activated the serine 270 phosphorylation of IRS correlates with JNK phosphorylation in INS-1 cells. Using Western blotting analysis, the levels of JNK phosphorylation were 3.33 fold increased and serine 270 phosphorylation of IRS was 1.17 fold increased in 33.3 G group compared to 11.2 G group (P<0.01). IGF-1 treatment inhibited phosphorylation of JNK and IRS. SP600125 treatment completely blocked JNK phosphorylation in 11.2 G group and reduced JNK phosphorylation by 90% in 33.3 G group. In addition, SP600125 treatment partly reduced serine 270 phosphorylation of IRS by 88.3% in 11.2 G group and 80% in 33.3 G group, the viability of INS-1 cells increased and the apoptosis decreased.CONCLUSION: The toxicity of chronic high glucose, which inhibits the cells viability and induces the cell apoptosis, might be related to suppress IRS signal by activating the JNK pathway. Blocking the JNK pathway might relieve the effect of glucose toxicity to the β cell function by improving the IRS signal pathway.

Background Experimental study showed that heparanase (HPA)is overexpressed in choroidal neovascularization,suggesting that it may play a role in the pathogenesis of angiogenesis.To certify HPA inhibitor suppress the formation and development of new blood vessel has an important significance for the treatment of choroidal neovascularization.Objective The aim of this study was to investigate the effect of HPA inhibitor on the proliferation of human umbilical vein vascular endothelial cell (UVEC) and the expression of HPA.Methods Hunan UVEC was primarily cultured and passaged and the third generation cells were used in the experiment.Phosphomannopentaose sulfate (PI-88) solution,a HPA inhibitor,was prepared with endothelial cell medium and the end concentrations were 400.00,200.00,100.00,50.00,25.00,12.50,6.25 mg/L respectively.The cells were treated with PI-88 solutions for 24,48 and 72 hours.The growth and proliferation of human UVEC were analyzed using MTT colorimetric assay at absorbance 570 nm.The expression of HPA in the cells was detected by immunochemistry in 48 hours after addition of PI-88.Results Cultured human UVEC showed the fusiform and polygon in shape.The A570 values of human CVEC were significantly different among various concentrations of PI-88 groups (F=2.721,P=0.053 ) and different action time (F=9.656,P =0.002).When PI-88 was administered for 24 hours,the A570 values of human UVEC were insignificantly altered in comparison with the one without PI-88 culture group (P＞0.05 ).However,in 72 hours after experiment,the A570 values were significantly declined as the PI-88 concentration was ＞50.00 mg/L ( P＜0.05 ).When PI-88 was administered for 48 and 72 hours,the A570 values of human UVEC were significantly higher than those of 24 hours in ＜50.00 mg/L groups (P＜0.01 ),but no statistical differences were seen in ＞100.00 mg/L groups among various time points (P＞0.05 ).HPA was intently expressed in the cytoplasm of human UVEC.However,at 48 hours after addition of ＞25.00 mg/L PI-88,the HPA expression was obviously weaker.Conclusions PI-88 can suppress the growth and proliferation of human UVEC at the dose-and time-dependent manner by downregulating the expression of HPA in the cells.

Objective To explore the effects of hypericin on inhibition of hepatitis 15 virus (HBV) replication.Methods The concentration gradient hypericin was added to HepG2.2.15 cell culture system and lamivudine was used as control.Enzyme-linked immunosorbent assay (ELISA) and Southern blot were used to examine HBsAg,HBeAg and HBV DNA level in the culture supernatant,respectively.Half inhibitory concentration (IC50) and half effective concentration (EC50) of hypericin were calculated.The effects of hypericin on HBV DNA polymerase were detected by 32p marked deoxy-ribonucleoside triphosphate as the substrate. Independent sample t-test and single factor analysis of variance were used to compare the data between two groups and among multiple groups.Results The inhibition rates of hypericin on HBsAg, HBeAg were enhanced with hypericin concentration increasing and those were higher than lamivudine control group when the concentration was higher than 0.5 μmol/L (t=-0.127,P＜0.05).Southern blot confirmed that hypericin was stronger in inhibition of HBV DNA (EC50=0.2 μmol/L) than lamivudine (t=-0.058,P＜0.05).Hypericin was non-toxic on HepG2.2.15 cells in the range of test with EC50 of 0.2 μmol/L and IC50 of 200 μmol/L.Hypericin did not act on HBV DNA polymerase which was quite different from lamivudine.Conclusions Hypericin can effectively inhibit HBV replication as well as antigen synthesis and is non-toxic on HepG2.2.15 cell.The anti-HBV target of hypericin is different from nucleos(t)ide analogues.

Objective: To investigate the efficacy of mifepristone in patients with Cushing's syndrome and the incidence of adverse reactions to provide basis for its clinical use.Methods: One case of Cushing's syndrome treated with mifepristone participated by pharmacist was analyzed.Results: Mifepristone could significantly improve hyperlipidemia and the complications of the patient, meanwhile, the risk of hypoglycemia should be paid attention and insulin therapy should be withdrawn timely.Conclusion: Mifepristone can significantly reduce blood glucose levels and improve the appearance of Cushing.At the same time, the level of serum potassium needs close monitoring to avoid the occurrence of hypokalemia.

Objective To evaluate the diagnostic value of combined detection of c-reactive protein(CRP)and cystatin C(Cys C) for early renal function impairment in gout patients,and to provide clinical reference.Methods Two hundred cases of gout patients were investigated and divided into experimental group and control group:experimental group patients(n=100)were with early renal function impairment and the control group(n=100)were without early renal impairment.The concentration of CRP and Cys C were detected with turbidimetric immunoassay method by OLYMPUS AU 400 automatic biochemistry analyzer.Results Serum CRP and Cys C concentrations of gout patients with early renal function impairment were significantly higher than those of gout patients without early renal function impairment(P<0.05).In the group of gout patients with early renal function impairment,the positive rate of serum CRP(75%)and Cys C(81%)had no significant difference(P>0.05).The positive rate of combination detection of CRP and Cys C was 92%,which was significantly higher than either CRP or Cys C(P<0.05).Conclusion The combination detec-tion of CRP and Cys C can improve diagnostis of early renal function impairment in gout patients.

Objective To study the effects of serum containing Psoralea on proliferation and melanogenesis in A375 human melanoma cells by serum pharmacological method. Method A375 human melanoma cells were cultured in vitro. Different concentrations of drug-contained serum were added to the culture medium during logarithmic growth phase. Melanocyte culture was determined by MTT and NaOH method. Results There was a significant difference between the control and the drug-contained serum test group on proliferation, and there was significant increase in 20% serum drug-contained serum on melanogenesis (P