This study was aimed to evaluate the subchronic toxicity of diosgenin in mice. A total of 80 mice were divided into 4 groups, which were 0 (control), 100, 200, and 400 mg·kg-1 by the random number table. Intragastric administration was given once a day for 90 days in the assessment of subchronic toxicity of diosgenin among mice. The observed indexes contained body weight, fur color, diet, feces, and etc. The detected indexes contained blood routine analysis, blood biochemistry and pathological examination. The results showed that compared with the control group, the body weights of mice in the male medication group were slight reduced. There were no significant hematologic and pathologic abnormalities. It was concluded that the subchronic toxicity of diosgenin with no observed adverse effect dose level was more than 400 mg·kg-1. The oral administration was relatively safe.

0.05). (4) The result of detoxincation in- vivo was not as good as that of antiserum.

Objective To probe the CT findings of menisci in degenerative osteoarthrosis of knees joint.Methods CT features of meniscus damage in 151 cases with degenerative osteoarthrosis of knees joint collected randomly from 2003~2005 were retrospectively analysed.Results CT features included:abnormal outline of meniscus or its edge to be coarse in 16 cases(10.5%),local hypodense in the meniscus in 107 cases(70.8%),gap sign in 9 cases(5.9%) and vacuum sign in 19 cases(12.5%).Conclusion CT scan is of benefit for evaluating the meniscus damage in degenerative osteoarthrosis of knees joint,which can be used to plan the mode of treatment.

Aim To investigate the effects of Ginkgo biloba extract(EGB)on tumor necrosis factor-alpha(TNF-?)in TNBS-induced colitis in rats and its mechanisms.Methods Colitis in rats was induced by colonic administration with 2,4,6-trinitrobenzene sulfonic acid(TNBS).Wistar rats were randomly divided into four groups,10 in each:normal group,model group,5-aminosalicylic acid(5-ASA group)and Ginkgo biloba extract group(EGB group).The levels of nitric oxide(NO),and glutathion peroxide(GSH-Px)were measured by biochemical methods.The expressions of TNF-? and nuclear factor kappaBp65(NF-?Bp65)in the colon tissues of colitis rats were detected by means of immunohistochemistry.The expressions of induce nitric oxide synthase(iNOS)in the colon tissues of colitis rats were detected by reverse transcription polymerase chain reaction(RT-PCR).The effects of EGB on colonic inflammation and macroscopic and histological damage were evaluated as well.Results Compared with the model group,treatment with EGB for 4 weeks significantly reduced colon macroscopic and histological damage,elevated the activities of GSH-Px and reduced the contents of NO,inhibited the protein expressions of TNF-? andNF-?Bp65,and decreased the mRNA levels of iNOS in the colon tissues of experimental colitis.Conclusions The probable mechanisms of EGB was that it ameliorated inflammatory injury in TNBS-induced colitis in rats by its reduction of TNF-?,NF-?Bp65 and iNOS levels.Then EGB could curb the inflammatory cascade effects of inflammatory mediators to protect ulcerative colitis.

AIM: To investigate the effect of 7-(4-methoxyphenyl)-5, 8a-diphenyl-1,2, 3, 7, 8, 8a-hexahydroimidazo[1,2-a] pyridine (TIP-6) on cell proliferation in human hepatoma cell line HepG2 and human normal hepatocyte cell line L02. METHODS: Typan blue assay was used to check the effect of TIP-6 on cell proliferation. The changes of cell morphology were observed by the phase contrast microscope. Flow cytometry (FCM) was used to check cell cycle. Autophagy and autophagic cell death were detected after acridine orange (AO) staining under fluorescent microscopy. Apoptosis was analyzed by Annexin V/7-AAD, DAPI staining and DNA ladder. NF-κB expression was detected with cellular immunochemistry. RESULTS: Cell proliferation inhibiting effect was appeared when treated with TIP6 from 60 μmol/L to 200 μmol/L, which was correlated with treated concentrations and time. The proliferation rates were just 12.10% and 18.75% (vs control) under 200 μmol/L 72 h in HepG2 and L02 respectively. Vacuolization were found more and more frequently with the increasing of TIP-6 concentrations and treated time prolonged. FCM results indicated that cells were blocked in G2/M phase, and more sensitive were found in HepG2 than L02. AO staining results indicated that the phenomenon of autophagy and autophagic cell death were occurred and appeared more potent with more TIP-6 and longer time treated. No apoptosis markers were found with Annexin V/7-AAD and DAPI staining, and no DNA ladders were found either, these indicated that TIP6 didnt induce apoptosis in these cells. NF-κB was found increased after treated with TIP-6, and the autophagic vacuole became more and more with the increasing of NF-κB protein, but the proliferation rates decreased at the same time. CONCLUSION: TIP-6 inhibited cell proliferation and induced autophagy and autophagic cell death in HepG2 and L02 cells. NF-κB activation may be involved in these effects.

Objective To report the experience on the ureteroscopic treatment of ureteral fibro-epithlial polyp by Holmium:YAG laser resection.Methods Of five cases,the polyp was located in the upper 1 third of the ureter in 2 cases,and in middle 1 third or ureter in 2 cases,in lower 1 third of ureter in 1 case.The length of the polyps ranged from 3 to 16 cm.Three patients presented with flank pain,4 with hematuria and 1 with hydronephrosis.Five patients underwent rigid ureteroscopic treat-ment.TUR was performed in the 2 polyp cases with prolapsing from the ureteral orifice.A Holmium:YAG laser was used to resect ureteral polyps.At the end of the procedure,a 7 F double-J ureteral stent was placed and indwelling for 6- 8 weeks.Results All operations were successfully done.The pathologic diagnosis were fibroepithelial polyp.Histologically,the polyps were composed of a central fibrovascular core surrounded by hyperplastic benign urothelium.The stroma of polyp consis-ted of fibrous connective tissue with minimal cellular infiltration,and occasional epithelial cell nests were seen.The average length of hospital stay was 3 d (range 2 to 5).The mean follow-up was 24 months (range 3 to 51),and all patients remained no recurrence.One patient developed a ureteral stricture 3 months after the treatment,and relieved by endoscopic incision by Holmium:YAG laser.Conclusion Endoscopic management of ureteral fibroepithelial polyps could be a treatment modality with minimal morbidity and good treatment results.

Objective To investigate the expressions of ABCG2 and V-ATPase in NSCLC and their expression rates in pathological classification, TNM stages and pathological grades and the expression correlation between ABCG2 and V-ATPase. Methods Expressions of ABCG2 and V-ATPase were accessed with EnVinsion immunohistochemistry in tumor samples from 92 NSCLC patients. The corresponding data was analyzed statistically. Results Expressions of ABCG2 and V -ATPase were found both in the lung adenocarcinoma and lung squamous cell cancer, and the difference between these two kinds of tumors was significant (P =0.003,0.000). ABCG2 expression was significantly different among TNM stages of lung adenocarcinoma (P=0.004) as well as among pathological grades of lung adenocarcinoma (P =0.028) and squamous cell carcinoma (P =0.000), while no significant difference was found among TNM stages of squamous cell lung carcinoma. The level of V-ATPase expression was associated with TNM stages of lung adenocarcinoma (P =0.026) and pathological grades of lung squamous cell carcinoma (P =0.002), however, among TNM stages of lung squamous cell carcinoma and pathological grades of lung adenocarcinoma, the difference was not significant. Additionally, the significant correlation was found between expression of ABCG2 and V-ATPase in all samples, adenocarcinoma and squamous cell carcinoma (P<0.001). Conclusion The significant correlation is found between expression of ABCG2 and V-ATPase, which indicate that they may co-work to participate in the mechanism of anticancer drug resistance.

STIM1 is recognized as an ER Ca2+ sensor of calcium release-activated calcium (CRAC) channel that is constructed by membrane protein Orai1, However, this regulatory system may also be regulated by other proteins. Reticulocalbin 2 (RCN2) was purified and identified from STIM1-Orai1 complex. Confocal microscopy revealed that RCN2 co-localized with STIM1 in ER before and after Ca2+ store depletion. Single cell [Ca2+]I measurements of RCN2 EF hands mutant showed slight influence on SOC electrophysiological characters. Furthermore, a novel collar form aggregation of RCN2 surrounding STIM1 clusters suggested that RCN2 potentially plays a role of structure maintenance in STIM1 clustering.

Objective To investigate the effects of glutamine (Gln) on proliferation and survival of small cell lung cancer H446 cells, and further to explore the potential mechanism. Methods The proliferation of H446 cells was detected at different time points (0, 24, 48, 72 and 96 h) by CCK-8 assay in Gln (+) group and Gln (-) group, and an optimal time was selected. Under the optimal time, Annexin V-FITC/PI staining, CellTiter-Glo? assay kit and flow cytometer were used to detect cell survival, cellular adenosine triphosphate (ATP) and reactive oxygen species (ROS) levels. Gln (-) group was used as the control group, under the condition of Gln deficiency, cellular ATP, cell proliferation and survival were detected after adding oxaloacetic acid (OAA) or dimethyl-α-ketoglutarate (DM-αKG). Gln (-) group was used as the control group, cellular ROS, cell proliferation, colony and survival were detected after treated with ROS scavenger N- acetyl cysteine (NAC). With different concentrations (0, 2, 5, 10 μmol/L) of glutaminase inhibitor BPTES, the optimal concentration was selected through the colony assay. The cellular ATP and ROS levels and cell proliferation were detected under the optimal concentration. H446 cells were treated with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES), ROS inducer hydrogen peroxide (H2O2) or the combination of them, and cell survival ratio was compared between two groups. Results The proliferation levels of H446 cells at 24, 48, which were decreased most significantly in 72 h in Gln (-) group. When 72 h was used as the optimal time, the cell survival ratio and ATP level were decreased, and the ROS level was increased, in Gln (-) group compared with those of Gln (+) group (P<0.05). There was a higher survival ratio in H446 cells in Gln (-)+OAA group and Gln (-)+DM-αKG group than that of Gln (-) group (P<0.05), but there were no significant differences in cell proliferation and ATP levels between Gln (-) group, Gln (-)+OAA group and Gln (-)+DM-αKG group. The ROS level was reduced, the cell proliferation, colony level and survival ratio were increased in Gln (-)+NAC group compared with those of Gln (-) group (P<0.05). Cloning assay showed that 10μmol/L was the optional concentration. Under this concentration, the proliferation and ATP level were decreased in Gln(+)+BPTES group (P<0.05), and cellular ROS level was up-regulated compared with Gln(+) group. The survival ratio was significantly lower in BPTES+H 2O2 group compared with BPTES (+) group or H2O2 (+) group. Conclusion Glutamine deficiency inhibits the proliferation and survival ratio of H446 cells through enhancing ROS level. BPTES and H2O2 show synergistically inhibitory effect on the survival of H446 cells.

[ ABSTRACT] AIM:To investigate the relationship between morphological changes of autophagy and apoptosis in the PC12 cells induced by oxygen-glucose deprivation and reoxygenation .METHODS:The PC12 cells were randomly di-vided into normal control group , oxygen-glucose deprivation and reoxygenation group , autophagy inhibitor group and auto-phagy activator group .The cells in oxygen-glucose deprivation and reoxygenation group , autophagy inhibitor group and au-tophagy activator group were exposed to reoxygenation (12 h) after 3 h of oxygen-glucose deprivation, and autophagy inhib-itor 3-methyladenine and autophagy activator rapamycin were added into the cells at the same time .Using transmission elec-tron microscope and monodansylcadaverine fluorescence staining , the morphological changes of autophagosome were ob-served.The apoptosis of the PC12 cells were analyzed by flow cytometry with Annexin V-FITC/PI staining and TUNEL method.RESULTS: Compared with normal control group , the numbers of autophagosomes and the apoptotic rates in-creased in oxygen-glucose deprivation and reoxygenation group (P<0.05).Compared with oxygen-glucose deprivation and reoxygenation group , the numbers of autophagosomes decreased obviously ( P<0.05 ) and the apoptotic rates increased markedly in autophagy inhibitor group (P<0.05).The numbers of autophagosomes increased obviously (P<0.05), the apoptotic rates decreased markedly ( P<0.05 ) , the autophagosomes became bigger in size , and autolysosomes was also found in autophagy activator group .CONCLUSION: Oxygen-glucose deprivation and reoxygenation induce autophagy in PC12 cells, and autophagy inhibits cell apoptosis to play a protective role .