Objective To investigate the effect of Ligu Capsules in preventing and treating menopausal osteoporosis in ovariectomized rats.Methods Rat models with osteoporosis were established by ovariectomy. Bone mineral density of femur,dry and wet bone weight and blood biochemical parameters were measured before and after treatment.Results Compared with the model group, Ligu Capsules increased femur bone mineral density,elevated the serum levels of Ca,P,Mg ,Zn and CT, decreased serum BGP level and hydroxyproline/creatinine(Hyp/Crea)ratio,increased dry weight, wet weight and ash weight of femur, as well as the bone length, bone volume, and bone diameter.Conclusion Ligu Capsules can prevent and treat osteoporosis in ovariectomized rats.

Objective To investigate the effects of silencing TSG-6 gene modified bone marrow mesenchymal stem cells (BMSC) transplantation on liver allotransplantation rejection in rats.Methods BMSC and KCs were isolated from rats and cultured in complete medium.We down-regulated TSG-6 expression of BMSC with lentiviral vectors carrying short hairpin RNA (LV3-shTSG-6).TSG-6mRNA and protein level of BMSC were tested respectively by quantitative real-time PCR and western blot analysis.After co-cuhured between BMSC and KCs,the expression of TNF-α and TSG-6 in cell supernatants were tested.Then,we established the orthotopic liver transplantation models in rats,the rats of each group were killed at 1 days,3 days and 7 days after operation.The serum levels of AST,ALT,TBIL,γ-GGT,TNF-α,IL-6 and IL-4 were tested with ELISA in each group,TSG-6mRNA and protein level of liver tissues obtained from each group were tested by quantitative real-time PCR and western blot analysis.The 1iver tissues of each group were stained with HE,then microstructure of liver tissues were observed under light microscope.The postoperative survival rates in other rats of each group were observed.Results The lentivirus transfection efficiency of mesenchymal stem cells was beyond 70 percent;After co-cultured between BMSC and KCs,the expression of TSG-6 in TSG-6-shRNA-BMSC + KCs group supernatant was significantly lower in different time point than that in BMSC + KCs group and TSG-6-NC-BMSC + KCs group (P ＜0.05),and the expression of TNF-α peak in BMSC + KCs group and TSG-6-NC-BMSC + KCs group supernatants were at 6 hours,which were significandy lower than those at 12 hours (P ＜0.05),but the expression of TNF-α in TSG-6-shRNABMSC + KCs group at 12 hours was significantly higher than those in BMSC + KCs group and TSG-6-NC-BMSC +KCs group (P ＜0.05);After transplantation,the serum levels of AST,ALT,TBIL,γ-GGT,TNF-α and IL-6 in TSG-6-shRNA-BMSC group was significantly higher than those in TSG-6-NC-BMSC group and BMSC group in different time point (P ＜ 0.05),but had no significant difference compared with PBS group;The serum levels of IL-4 in TSG-6-shRNA-BMSC group,TSG-6-NC-BMSC group and BMSC group was significantly higher than that in PBS group(P ＜ 0.05),but TSG-6-shRNA-BMSC group compared with TSG-6-NC-BMSC group and BMSC group,the serum levels of IL-4 was significantly lower (P ＜ 0.05);In pathological changes,we found that the degree of liver rejection in TSG-6-shRNA-BMSC group and PBS group were seriously obvious,and were graded Ⅱ-Ⅲ with Banff schedule;Comparation of postoperative survival time in each group,TSG-6-shRNA-BMSC group and PBS group were (16.6 ±4.6) d and (15.4 ± 6.7) d respectively,which were signifcantly lower than those in TSG-6-NCBMSC group (69.6 ± 28.1) d and BMSC group (69.2 ± 28.2) d (P ＜ 0.05).Conclusion Transplantation of BMSC secreted TSG-6 could,to some extent,mitigate acute liver transplantation rejection.

AIM: To examine the apoptotic pathway of norcantharidin (NCTD)-induced HeLa cells death. METHODS: MTT, photomicroscopical observation, DNA agarose gel electrophoresis, LDH release and Western blot analysis were used. RESULTS: NCTD induced HeLa cells apoptosis and the apoptosis was partially reversed by the inhibitors of caspase-family (-3, -8, -10). The activities of caspase-3, -8 and -9 were significantly increased after treated with NCTD. The expression of the inhibitor of caspase-3 activated DNase (ICAD) was decreased in a time dependent manner. CONCLUSION: NCTD induces HeLa cells apoptosis through activating caspase pathways.

AIM: To observe the changes of interleukin1 receptor associated kinase-4(IRAK-4) in ischemia/reperfusion(I/R) liver pretreated with lipopolysaccharide(LPS) and to explore the protective mechanisms of LPS pretreatment against hepatic I/R injury.METHODS: Male Sprague-Dawley rats,weighing 240-280 g,were divided into three groups: control,ischemia/reperfusion group(I/R group) and LPS-pretreated group(LPS group).On the first day,LPS group received 0.1 mg/kg LPS via the tail vein,followed by 0.5 mg/kg on the 2nd,3rd,4th and 5th day.I/R group received the equivalent volumes(0.5 mL) of sterile PBS.Experiments of I/R injury was induced by temporary ischemia of the left lateral liver lobe for 90 min followed by 3 h reperfusion on 2 days after the last LPS treatment.At 0 min,60 min and 180 min after reperfusion,the expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blotting.The activity of NF-?B and the serum TNF-? level were also detected by ELISA.RESULTS: Although the level of IRAK-4 gene and protein were higher in the LPS group than that in I/R group and control group(P0.05) at 0 min after reperfusion.However,all those indexes were evidently lower in the LPS group than those in I/R group(P

A terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay was used to determine that apoptosis causes HeLa cell death induced by pseudolaric acid B. The c-Jun N-terminal kinase (JNK) inhibitor SP600125 decreased p53 protein expression during exposure to pseudolaric acid B. SP600125 decreased the phosphorylation of p53 during pseudolaric acid B exposure, indicating that JNK mediates phosphorylation of p53 during the response to pseudolaric acid B. SP600125 reversed pseudolaric acid B-induced down-regulation of phosphorylated extracellular signal-regulated protein kinase (ERK), and protein kinase C (PKC) was activated by pseudolaric acid B, whereas staurosporine, calphostin C, and H7 partly blocked this effect. These results indicate that p53 is partially regulated by JNK in pseudolaric acid B-induced HeLa cell death and that PKC participates in pseudolaric acid B-induced HeLa cell death.
Tumor Suppressor Protein p53/metabolism/*physiology ; Protein Kinase C/metabolism ; Phosphorylation ; JNK Mitogen-Activated Protein Kinases/*physiology ; Humans ; Hela Cells ; Diterpenes/*pharmacology ; DNA Fragmentation/drug effects ; Cell Death/*drug effects ; Anthracenes/pharmacology

Pseudolaric acid B was isolated from Pseudolarix kaempferi Gordon (Pinaceae) and was evaluated for the anti-cancer effect in HeLa cells. We observed that pseudolaric acid B inhibited cell proliferation and induced apoptosis in a time- and dose-dependent manner. HeLa cells treated with pseudolaric acid B showed typical characteristics of apoptosis including the morphological changes and DNA fragmentation. JNK inhibitor, SP600125, markedly inhibited pseudolaric acid B-induced cell death. In addition, Bcl-2 expression was down-regulated while Bax protein level was up-regulated. Caspase-3 inhibitor, z-DEVD-fmk, partially blocked pseudolaric acid B-induced cell death, and the expression of two classical caspase substrates, PARP and ICAD, were both decreased in a time- dependent manner, indicative of downstream caspase activation.
Anthracenes/pharmacology ; *Apoptosis ; Caspases/antagonists & inhibitors/*metabolism ; Cell Proliferation/drug effects ; Cysteine Proteinase Inhibitors/pharmacology ; Diterpenes/*pharmacology ; Down-Regulation ; Enzyme Activation ; Hela Cells ; Humans ; JNK Mitogen-Activated Protein Kinases/drug effects/*metabolism ; Oligopeptides/pharmacology ; Protein Kinase Inhibitors/pharmacology ; Proto-Oncogene Proteins c-bcl-2/metabolism ; Signal Transduction/*drug effects ; Up-Regulation