OBJECTIVE:To establish a microbial limit test method for Naoliqing capsules and validate the methodology.METHODS:In accordance with Chinese Pharmacopoeia 2005th edition,a verification test on the microbial limit test mothod of Naoliqing capsules was carried out using escherichia coli,bacillu subtilis,staphy lococcus aureus,aspergillus niger and candida albicans.RESULTS:The recovery rates of the above different test bacteria were all above 70%,and the recovery rates of 5 test bacteria in diluting agent were all above 70%.CONCLUSION:The routine method is applicable for microbial limit test of Naoliqing capsules.

Fibrodysplasia ossificans progressiva(FOP) is a rare debilitating genetic disorder characterized by malformation of the great toes during embryonic skeletal development as well as progressive heterotopic ossification postnatally. There is no known definite treatment. Here presented a review of the recent advances in the potential FOP therapies.

To explore the detection of GNAS1 gene mutation from peripheral blood and non-involved bone marrow in patients with fibrous dysplasia of bone(FD). Both peripheral blood and non-involved bone marrow collected from six FD patients were used for genetic diagnosis by next generation sequencing and direct sequencing respectively. Two patients were found to have common gene mutation(positive) by next generation sequencing, accounting for 33.3% of total six people: p.R201H(CGT>CAT) was detected in both peripheral blood and bone marrow of patient No.3, and p.R201C(CGT>TGT) was detected in both tissues of patient No.6. While direct sequencing only found p.R201H(CGT>CAT) mutation in peripheral blood and bone marrow of patient No.3. Detection of GNAS1 gene mutation may contribute to the diagnosis of FD. Hot spot mutations of GNAS1 are present in peripheral blood and non-involved bone marrow samples from some Chinese FD patients, but the diagnostic value and sensitivity of GNAS1 gene mutation in non-involved tissues of FD patients is low by next generation sequencing or direct sequencing.

Objective To explore the correlation between simple thyroid peroxidase antibody (TPOAb) positive (higher than the normal reference value) and the adverse pregnancy outcomes;and to discuss the necessity of clinical intervention on TPOAb positive and the characteristic of gestational thyroid peroxidase antibody changes.To study wether clinical intervention affects its direction.Methods Subjects were selected from TPOAb positive pregnant women during June 2012 to February 2015.They were registered regularly for thyroid function during pregnancy and 6 weeks postpartum,and timely treated for abnormal thyroid function.They were grouped according to the thyroid function:Group 1 (n =122),simple TPOAb positive group;Group 2 (n =67),patients with elevated TSH and positive TPOAb who accept the levothyroxine (L-T4) intervention;Group 3 (n =120),TPOAb negative healthy pregnant women.They were regularly tested for thyroid function during pregnancy,and with follow-up until 6 weeks postpartum.Results When Group 1 compared with Group 3:the risk of premature rupture of membranes,abnormal amniotic fluid,and fetal distress rate increased,the relative risk (RR) were 3.066,4.782,2.605,all P ＜0.01,while the gestational diabetes,gestational hypertension,preterm birth,abnormal gestation and birth showed no statistical differences (P＞0.05).When Group 2 compared with Group 3,the risk of premature rupture of membranes,abnormal amniotic fluid,and fetal distress increased (RR were 3.830,5.537,3.600 respectively,all P ＜ 0.01).While gestational diabetes,gestational hypertension,preterm birth,abnormal gestation and birth did not show any statistical difference (P＞0.05).Compared with Group 2,Group 1 had no statistical difference between the above observational parameters.The levels of TPOAb from stage T1 (the first trimester) to T3(the third trimester) showed a overall downward trend in Group 1 and Group 2,rise again six weeks after child birth.Comparison of the two groups (Group 1 and Group 2),the median value during this period did not reveal any significantly statistical differences (P＞ 0.05).Conclusion In simple TPOAb positive pregnant women,premature rupture of membranes,abnormality in amniotic fluid,and fetal distress were significantly higher than those in healthy pregnant women;even with thyroid function correction the TPOAb positive pregnant women still cannot reduce the adverse pregnancy outcomes.Levels of TPOAb from T1 to T3 presented the overall downward trend and to rise again six weeks after pregnancy.L-T4 intervention for TPOAb positive pregnancy did not show any significant effect.

Objective Isobaric tag for relative and absolute quantitation (iTRAQ) was applied for the quantitative analysis of serum protein in psoriasis vulgaris patients with damp heat syndrome and healthy volunteers to explore the differentially expressed proteins, thus to reveal the nature of damp heat syndrome from the proteomics level.Methods The serum from 15 psoriasis vulgaris patients with damp heat syndrome and 10 healthy volunteers was purified and treated with removal of abundance. After separation with sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and hydrolysis of peptide specific iTRAQ marker enzyme, the differentially expressed proteins of serum samples were identified by tandem mass spectrometry. Results A total of 787 proteins were identified, and 718 proteins had the functional annotation showed by gene ontology ( GO) in psoriasis vulgaris pa tients with damp heat syndrome. A total of 651 differentially expressed proteins were identified by compared with healthy volunteers ( P<0.05) , and markedly significant difference was shown in 418 proteins ( P<0.01) . Conclusion Psoriasis vulgaris patients with damp heat syndrome have differentially expressed proteins which are different from those in the healthy volunteers, but which kind of protein being specific for damp-heat psoriasis vulgaris as well as the relation between the damp heat symptoms and proteins still need to be further studied.

AIM: To explore the effect and the mechanism of sulphated heparin on the proliferation and the apoptosis of human hepatocellular carcinoma cells. METHODS: The human hepatocellular carcinoma cell line (HepG-2) was used to identify the expression of ras gene protein and to study the effect of sulphated heparin on proliferation and the apoptosis in vitro . RESULTS: The sulphated heparin downregulated the ras protein expression and inhibited the cell growth in HepG2 cells. In the presence of sulphated heparin, the apoptosis rate of HepG2 increased. CONCLUSION:The data suggest that the effects of sulphated heparin on the proliferation and the apoptosis of the human hepatocellular carcinoma cell are correlated with the signaling transduction mediated by ras gene protein.

In order to improve the Chinese medical students' ability to gain the latest medical information and adapt the development of modern medical education,we carried out bilingual teaching of Urology in undergraduate students.Based on students English level,we teach students step by step in accordance with their aptitude and build English study group.The practice shows that we will achieve better teaching effect if we sum up our experiences constantly.

Objective To assess the coordinated regulation and the molecular mechanisms of 17β-estradiol and 1,25-dihydroxyvitamin D3 [ 1,25-( OH ) 2 D3 ] on the proliferation and differentiation of osteoblastic MC3T3-E1 cells.Methods MC3T3-E1 cells were cultured in phenol-red free α-MEM medium supplemented with 10％ FBS,MTT assay was performed to determine the effects of 17β-estradiol and 1,25-( OH )2 D3 on MC3T3-E1 cells proliferation.After cells were treated with different agents,cell cycle related genes [ cyclin E,proliferation cell nuclear antigen ( PCNA ),and cyclin-dependent kinase inhibitor 2b ( Cdkn2b ) ] and markers of osteoblastic differentiation [ type Ⅰ collagen ( COL Ⅰ ),alkaline phosphatase ( ALP),osteopontin ( OPN ) ] were detected with SYBR green-based quantitative PCR.ALP activity was detected with BCIP/NBT method.Results 17β-estradiol could promote proliferation of MC3T3-E1 cells,which was accordant to its ability to increase cyclin E and PCNA and to inhibit Cdkn2b mRNA expression in MC3T3-E1 cells.However,1,25-( OH)2D3 had no effect on the proliferation of MC3T3-E1 cells and also did not enhance the proliferation of MC3T3-E1 cells stimulated by 17β-estradiol.On the other hand,17β-estradiol promoted the gene expression of differentiation markers Col Ⅰ,ALP,and OPN,and 1,25-(OH) 2 D3 synergistically increased the expression of these genes with 17 β-estradiol.Conclusion As two of the most important hormones which regulate bone metabolism,estrogen and vitamin D may coordinately promote osteoblast differentiation,but may not regulate osteoblasts proliferation synergistically.

Objective The aim of this study was to explore the effect of low density lipoprotein ( LDL) on the proliferation and differentiation of MC3T3-E1 osteoblasts, as well as the expression of low density lipoprotein receptor-related protein 5(LRP5) and dickkopf-1(DKK1) mRNA of MC3T3-E1 osteoblasts. The possible mechanisms of the treatment of atorvastatin on LDL expression in MC3T3-E1 osteoblasts were also investigated. Methods Proliferation, osteocalcin expression, LRP5, and expression of DKK1 mRNA of MC3T3-E1 with interaction of LDL at 0. 05, 0. 10, 0. 20 mg/ml levels after 24 h, 48 h, 72 h were detected by CCK8, ELISA, and fluorescence quantitative PCR. Furthermore, proliferation, osteocalcin expression, LRP5 and DKK1 mRNA of MC3T3-E1 after the treatment of atorvastatin of 10-6 mol/L and 10-5 mol/L were also be studied, respectively. Results The effect of LDL on proliferation, expression of osteocalcin and expression of LRP5 and DKK1 mRNA in MC3T3-E1 osteoblasts was the most obvious under LDL with 0. 20 mg/ml level. Under that level, atorvastatin (10-6 mol/L or 10-5 mol/L) was able to make the proliferation of MC3T3-E1 osteoblasts in 48 h and 72 h be decreased, while significantly caused upregulation of osteocalcin, LRP5 mRNA expression; and down regulated DKK1 mRNA expression ( all P<0. 05). Conclusions Atorvastatin can reduce the inhibitory effect of LDL on the proliferation and differentiation of osteoblasts. The mechanisms of atorvastatin on osteoblasts are possibly related to the osteoblast proliferation and expression of LRP5 mRNA and DKK1 mRNA of osteoblasts of wnt signal pathway.

Objective To explore the detailed underlying molecular and signaling mechanisms in the effects of icariin on bone formation by an in vitro cell model. Methods The proliferation of MC3T3-E1 osteoblast-like cells was evaluated by MTT, and gene expression of cell cycle related proteins in MC3T3-E1 cells after icariin treatment was detected by real-time PCR. The phosphorylation of MAPK signals, including ERK, P38, and JNK was determined by Western blot, and then the inhibitors of MAPK signals were used to treat cells with icariin alone or together to determine the role of MAPKs in the process of icariin treatment on MC3T3-E1 cell proliferation. Alkaline phosphatase and Alizarin red staining were used to detect the formation of mineralization nodules, and gene expressions of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in osteoblasts after being treated by icariin were evaluated by real-time PCR. ICI182780, and nilutamide was used to decide the participation of estrogen and androgen receptor signals in the process of icariin treatment on the differentiation and mineralization of MC3T3-E1 cells. Results Treatment with icariin promoted MC3T3-E1 cell growth in a time- and dose-dependent manner. This treatment also revealed that icariin increased the expression of mRNAs encoding both cyclin E and PCNA, positive regulators of cell growth, but decreased levels of mRNAs encoding Cdkn2b, a negative regulator of cell cycle progression. When MC3T3-E1 cells were cultured in a differentiated condition, icariin enhanced mineralized nodule formation and increased the expression of mRNAs encoding alkaline phosphatase, type Ⅰ collagen, and osteocalcin. Treatment with icariin significantly induced phosphorylation of both ERK and JNK and this phosphorylated effect occurred very rapidly within 5 minutes and reached peak at 15 minutes. Furthermore, the stimulated effects of icariin on proliferation and gene expression of cyclin E, PCNA, and Cdkn2b in MC3T3-E1 cells were dramatically attenuated by treatment with both U0126 and SP600125, inhibitors of MAPKs. Interestingly, such stimulating effects of icariin were at least partly reduced by treatment with ICI182780, an inhibitor of estrogen receptor. Icariin induced mineralized nodule formation and gene expression of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in MC3T3-E1 cells were also partly reduced when the cells were treated with ICI182780. Conclusions Our findings indicate that the anabolic effect of icariin on bone formation is, at least partly, mediated through the MAPK signaling pathway in order to modulate osteoblast proliferation and differentiation.