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 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

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

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.

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 clinical efficacy of rhubarb acupoint sticking joint shuganjieyu diet in the treatment of chronic functional constipation patients with depressive anxiety.Methods 120 patients of chronic functional constipation with depression anxiety were selected,they were randomly divided into control group and obser-vation group,60 cases in each group.The patients in the control group were given promoting gastrointestinal motility drugs mosapride capsules,and orally given laxatives rhubarb with sodium bicarbonate.The patients in the observation group were treated with topical rhubarb acupoint,and orally given medicated shuganjieyu diet.7 days for a course of treatment,the patients were treated for four courses.The clinical efficacy of the two groups,defecation interval and fecal nature,SAS,SDS scale score changes and quality of life (PAC -QOL)scale score changes before and after treatment and adverse reactions during treatment were recorded.Results The total effective rate of the observation group was 90.00%,which of the control group was 91.67%,the difference between the two groups was not significant (χ2 =0.32,P >0.05).Before treatment,the bowel movement and stool ratings between the two groups had no statis-tically significant differences (t =0.23,0.33,all P >0.05).After treatment,the two points of the two groups were significantly decreased,but the differences between the two groups were not significant (t =0.18,0.41,all P >0.05).Before treatment,the SAS,SDS scale scores between the two groups had no statistically significant differences (t =0.78,0.69,all P >0.05).After treatment,the SAS,SDS scores of the two groups were significantly decreased, which of the observation group decreased more significantly than those of the control group (t =4.18,4.72,all P <0.05).Before treatment,the PAC -QOL scale scores for each dimension level had no statistically significant differ-ences (t =0.16,0.21,0.42,0.20,all P >0.05).After treatment,the dimensions of the two groups showed a decline trend,and those of the observation group decreased more significantly than the control group (t =2.10,3.56,2.76, 2.34,all P <0.05).In the control group during the treatment,2 cases of diarrhea,three cases of dry mouth,the patients were improved after given symptomatic treatment.No serious adverse event occurred during therapy in the observation group.Conclusion Rhubarb acupoint joint shuganjieyu diet in the treatment of chronic functional constipation patients with depression anxiety has significant clinical effect,which can significantly improve patients'depression and anxiety scores,quality of life,and without significant adverse reactions,it is worthy of clinical applica-tion.

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

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