BACKGROUND: In recent years, it has been confirmed by a variety of ways that myoblasts can differentiate into osteoblasts under the induction of recombinant human bone morphogenetic protein 2 (rhBMP-2).OBJECTIVE: To explore the mineralization reaction during the osteogenic differentiation of myoblasts under the induction of recombinant rhBMP-2 and the feasibility of osteogenic phenotype expression by in vitro induction. METHODS: Myoblasts were isolated and harvested from neonatal Wistar rats using differential velocity adherent technique and trypsinization method. After in vitro culture, purification and identification, myoblasts at passage 3 were induced by a medium containing rhBMP-2 for 21 days. Myoblasts in the control group were cultured in vitro in complete medium without rhBMP-2 for 21 days. RESULTS AND CONCLUSION: After rhBMP-2 induction, myoblast proliferation gradually slowed down. A small quantity of opaque secretory granules were found in the cytoplasm on day 8 after induction; the number of opaque secretory granules increased on day 14 after induction; and a great quantity of opaque secretory granules were found in the cytoplasm on day 21 after induction while the myoblasts without induction fused into contractile myotubes. The alkaline phosphatase activity of the induced myoblasts increased as time extended; myoblasts reacted positively in the alkaline phosphatase staining, immunochemical staining for type Ⅰ collagen and calcium node staining on day 21 after induction. These findings suggest that mineralization reaction is found in rat myoblasts by rhBMP-2 induction and myoblasts can differentiate into osteoblasts under certain inducing conditions in vitro.

Objective To investigate the clinicopathologieal features of serous microcystic adenoma of the pancreas. Methods Clinicopathological analysis, histochemieal and immunohistochemical studies were performed in 12 cases of serous microcystic adenoma of the pancreas with follow-up. Results The patients included 9 females and 3 males who ranged in age from 35 to 70 (mean=51.5). Seven patients were discov-ered by check-up incidentally and 5 for the symptoms as upper abdominal uncomfortable or pain. Tumors varied in size from 2.0 cm to 13 cm in maximal diameter(mean=6.0 cm), and all had a multicystic appear-ance. Histologically, all tumors were composed of numerous microcysts lined by a single layer of cuboidal or flattened cells. The tumor cells contained clear cytoplasm with glycogen and centrally located round nuclei mostly uniform without any atypia. Mitoses were very rare or absent. Histochemically, the tumor cells were positive for PAS and negative for AB-PAS. Immunohistochemieally, the tumor cells revealed diffuse positivi-ty of cytokeratins and epithelial membrane antigen, whereas CEA, CgA, Syn, CD34, Vimentin, P53 and S-100 protein were negative. Follow-up in 10 patients (range from 12 to 71 months) showed no recurrence of serous cystadenomas. Conclusion Serous microcystic adenoma of the pancreas is an extremely rare tumor. It is often non-symptomatic. Its diagnosis is mainly dependent on its unique morphological immunohistochemical features. It's biology behavior is necessary to study.

Objective To set up the filter mating method of plasmid conjugation experiment for Corynebacterium.Methods The first plasmid conjugation between different genus was performed by filter method.Three strains of highly erythromycin-resistance Enterococcus faecalis isolates[minimal inhibitory concentration(MIC)of erythromycin>256 mg/L,MIC of levofloxacin≤8 mg/L] which screened by agar dilution method were used as donators and 3 strains of Corynebacteria isolates(MIC of erythromycin≤32mg/L,MIC of levofloxacin=128 mg/L)were used as acceptors.In the second conjugation between the same genus but in different species,three Corynebacterium xerosis strains(MIC of erythromycin>256 mg/L,MIC of ceftazidime=16 mg/L)which were conjugants of the first conjugation were used as donators and 6 strains of Corynebacteria isolates(MIC of erythromycin≤32 mg/L,MIC of ceftazidime 128 mg/L)were used as acceptors.Results In the first conjugation between different genus,the plasimid conjugations were done for 9 times between 3 donators and 3 acceptors,4 of which gained the phenotype and genotype of resistance and the transfer rate was 44%.In the second conjugation experiment,the plasmid conjugations were done for 18 times between 3 donators and 6 acceptors,7 of which gained the resistant phenotype and the transfer rate was 39%.Erythromycin-resistance was transferred from the donators to the conjugants in varying degrees.The resistance was transferred from Enterococcus faecalis to Corynebacteria and then to other Corynebacteria which were of the salne genus but of different species.Conclusion Filter mating conjugation method could be used to study the mechanisms of resistance transmission of Corynebacterium.

Objective To explore the expression of mismatch repair gene (MMR) (MLH1,MSH2,PMS2 and MSH6) in endometrioid adenocarcinoma,and to analyze its clinical pathological significance.Methods Inmunohistochemical EnVision method was used to detect the expressions of DNA mismatch repair proteins (MLH1,MSH2,PMS2 and MSH6) in 101 cases of endometrial adenocarcinoma.The phenotypes of tumor microsatellite instability (MSI) and microsatellite stable were determined,and its relationship with onset age,differentiation,depth of myometrial invasion,lymphatic metastasis and prognosis of tumor were analyzed.Results There were 67 cases of hysterectomy and 34 cases of endometrial biopsy in 101 cases of endometrial adenocarcinoma.52 cases of surgical specimens and 14 cases of biopsy specimens were included in 66 cases that had complete follow-up data.Rates of negative expression were as follows:MLH1 31.6 ％ (32/101),MSH228.7 ％ (29/101),PMS2 16.8 ％ (17/101),MSH6 8.9 ％ (9/101).MSI phenotype 53.5 ％ (54/101),MSS phenotype 46.5 ％ (47/101).Univariate analysis showed that prognoses of MSI-L&MSS group (34 cases) were better than those of MSI-H group (18 cases) in hysterectomy organization (P =0.041).The differences between depth of myometrial invasion,degree of differentiation,age and prognosis of endometrial adenocarcinoma were statistically significant (P ＜0.05).Cox multivariate analysis showed that differences between depth of myometrial invasion,age and prognosis of endometrial adenocarcinoma were statistically significant (P =0.034,P =0.009),while there was no significant difference between MSI-L&MSS group and MSI-H group in prognosis (P ＞ 0.05).Conclusions MSI is one of the molecular events that occur in the endometrioid adenocarcinoma.Age and depth of tumor invasion are independent prognostic factors of endometrioid adenocarcinoma.In view of inconsistency between univariate analysis and multivariate analysis,whether MSI can be used as deterministic accordance for endometrioid adenocarcinoma prognostic evaluation requires further verification.

OBJECTIVE:To establish a HPLC method for the determination of paracetamol,caffeine and aspirin in compound paracetamol tablets.METHODS:The chromatographic column was ODS,the mobile phase was methanol-4.2% glacial acetic acid solution(3∶7)with a flow rate at1.0ml/min;the detecting wavelength was 275nm,the sample size was 20?l and the column temperature was 25℃.RESULTS:The linear ranges of paracetamol,caffeine and aspirin were 16.08～428.80?g/ml(r=0.9981),3.42～91.20?g/ml(r=0.9988) and 23.87～769.92?g/ml(r=0.9999) respectively;The average recovery were 99.2%(RSD=0.23%),100.4%(RSD=0.63%)and 99.3%(RSD=0.11%)respectively.CONCLUSION:This method was simple,fast and accurate,which can be used for the content determination of compound paracetamol tablets.

The research hot spots and development in domestic neurology were analyzed by co-words analysis, visual analysis and the combination of them, which showed that the co-occurrence relation between high frequency words could display the structure and hot spots of subject headings in neurology, analysis of the cluster evolution and position could dynamically reveal the hot spots and development in neurology, and comprehensive analysis of words and their classes could directly and overall demonstrate the hot spots and development in neurology.

Rheumatoid arthritis (RA) is a common systemic autoimmune disease characterized by chronic inflammation of joints,bone and cartilage erosion,synovial hyperplasia,and the pathogenesis of RA is not clear.γδT cells are a new kind of phenotype and function T lymphocyte subsets,which mainly distribute in the mucosal and epithelial tissue and account for 1％-10％ of the total T cells in the peripheral blood,and bridge innate and adaptive immunity.γδT cells play an important role in the pathogenesis of RA by the functions of antigen-presenting capacity,secretion of proinflammatory cytokines,immunomodulatory effects,and auxiliary function for B cells.

Objective To investigate the role of c-Jun N-terminal kinase (JNK) signaling pathway in hyperoxia-induced expression of connective tissue growth factor (CTGF) in A549 cells.Methods The cultured A549 cells were seeded in 6-well culture plates at a density of 1 × 105 cells/ml (0.5 ml/well,24 wells in total) and were randomly divided into 4 groups (n =6 each) using a random number table:control group (group C);95％ oxygen group (group 95％ O2);room air plus SP600125 (JNK inhibitor) group (group C+SP);95％ oxygen plus SP600125 group (group 95％ O2+SP).SP600125 5 μmol/L was added to each well in C+SP and 95％ O2+SP groups.The A549 cells were incubated for 48 h in an incubator filled with room air (C and C+SP groups) or 95％ oxygen (95％ O2 and 95％ O2+SP groups).The expression of phosphorylated JNK (p-JNK) and CTGF mRNA was determined using Western blot and realtime reverse transcriptase polymerase chain reaction,respectively.Results Compared with group C,no significant change was found in the expression of p-JNK and CTGF mRNA in group C+SP (P＞0.05),and the expression of p-JNK and CTGF mRNA was significantly up-regulated in group 95％ O2 (P＜ 0.05).Compared with group 95％ O2,the expression of p-JNK and CTGF mRNA was significantly down-regulated in group 95％ O2+SP (P＜0.05).Conclusion JNK signaling pathway activation is involved in up-regulation of hyperoxia-induced CTGF mRNA expression in A549 cells.

OBJECTIVE:To explore the safety and effects of liver function in patients with liver cirrhosis following virus B hepatitis(called“hepatitis B”for short)on drug half-life and analgesic effect of target controlled infusion of remifentanil. METH-ODS:100 patients with liver cirrhosis following hepatitis B underwent liver and gallbladder surgery under selective general anesthe-sia were collected from our hospital and divided into group A(mild abnormal liver function)and group B(severe abnormal liver function,3 cases withdrew from the test and 47 cases completed the test),with 50 cases in each group,according to Child-Pugh grading of liver function. Both group were given phenobarbital sodium 0.1 g+scopolamine 0.3 mg intramuscularly 0.5 h before oper-ation;midazolam 0.04 mg/kg+propofol 1.5 mg/kg+atracurium 0.6 mg/kg intravenously;target controlled infusion of Remifentanil hydrochloride for injection during operation with 0.125-0.250 μg/(kg·min). The distribution half-life and the elimination half-life of remifentanil were determined, and temperature pain perception threshold (tPDT) and electrical pain perception threshold (ePDT) were measured immediately after the operation;the occurrence of ADR was observed. RESULTS:The distribution and elimination half-life of remifentanil were (4.52 ± 1.25)min and(24.64 ± 1.30)min in group A and (4.68 ± 1.31)min and(25.45 ± 2.08)min in group B respectively,there was no statistical significance between 2 groups(P>0.05). tPDT and ePDT of group A were(8.88± 1.66)mA and(1.54±0.09)mA respectively,and those of group B were(9.16±1.58)mA and(1.34±0.15)mA,there was no sta-tistical significance between 2 groups (P>0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:The abnormal liver function of patients with liver cirrhosis following hepatitis B have no significant effect on drug half-life and analgesic effect of remi-fentanil with good safety.

Objective To seek the effect of SGK1 on the protection and prognosis of cerebral ischemia reperfusion. Methods Animal model of rats was used to construct them into experimental group(A)and control group(B). The impact of cerebral ischemia reperfusion on the hippocampus neuron cell apoptosis was simulated by making the overexpression of SGK1 and using PI3K inhibitors LY294002 to deal with animal models. Results The overexpression of SGK1 could reduce the neuron cell apoptosis caused by cerebral ischemia reperfusion to a certain extent,reverse the expressions of Cleaved aspas 3 protein,pro-apoptotic protein Bax and inhibitor of apoptoasis protein Bcl-2 through the PI3K/Akt-mediated signaling pathway. Conclusion SGK1 can protect the tissues with ischemia-reperfusion,which may provide the biological evidence for future clinical applications.