Objective To construct a sequence tag library of HepG2 cells by serial analysis of gene expression (SAGE). Methods Total RNA of HepG2 cells was extracted using a Trizol agent. The double strand cDNA were reverse-translated from mRNA which was isolated from total RNA. According to the protocol of SAGE, a sequence tag library of HepG2 cells was constructed. The transcript, which was represented by each tag, was analyzed by consulting the GenBank and UniGene, and the abundance of the tags was analyzed by SAGE analysis software. Results A total of 14 181 SAGE tags out of 15 586 tags were steadily and effectively amplified and the lengths of the tags were between 200-3000 bp. After sequence analysis of the tags, 2023 specific genes were revealed. Conclusions The use of improved reverse transcription agent kit may guarantee obtaining the full length cDNA, which provides materials for the construction of SAGE library. SAGE is a powerful method to investigate the gene expression profile, and it establishes a foundation for gene researches.

Objective To study the proliferation and differentiation patterns of hepatic stem cells from mice cultured in vitro isolated identify their characteristics.Methods Mice were divided into 5 groups according to the pregnant (embryo days,ED13,ED15 and ED18) or born age (day0 and 3).Hepatic stem cells were isolated and cultured in vitro.The amount of the stem cells as well as their growing situation and differentiation pattern were observed and compared among different groups and markers of stem cells (CD117,CD90.1,CD49f,c-Met),hepatic cells(AFP) were used to identify the cultures.Resulls The cells with best situation of growing as colonies were obtained from ED15 group.Their expression of specific markers suggested that they were hepatic stem cells.The stem cells isolated from ED15 mice in subculturing proliferated in line pattern and differentiated in reverse line pattern.The expression of AFP varied as normal distribution as cell differentiation development.Conclusion Most cells have characteristics of hepatic stem cells isolated from fetal liver and the number of these cells decreases gradually as embryo duration prolongs.The hepatic stem cells proliferate in line pattern and differentiate into hepatic cells after in vitro culture.

ObjectiveTo explore the early diagnostic value of pro-adrenomedullin (pro-ADM) in sepsis. Methods A prospective study was conducted. Eighty-two patients with acute infection admitted to Department of Emergency of Shanxi Medical University Second Hospital from April 2013 to March 2014 were enrolled. According to the diagnostic criteria of sepsis, the patients with acute infection were divided into ordinary infection group [infection without systemic inflammatory response syndrome (SIRS),n = 25] and sepsis group (infection combined with SIRS, n = 57). According to degree of severity of sepsis, the latter group was subdivided into three subgroups: sepsis group (n = 22), severe sepsis group (n = 27) and septic shock group (n = 8). Twenty-four healthy persons were included to serve as healthy control group. The venous blood from all the research objects in hospital was collected within 24 hours. The levels of pro-ADM and procalcitonin ( PCT ) were determined by enzyme linked immunosorbent assay (ELISA), and acute physiology and chronic health evaluationⅡ (APACHEⅡ) score was recorded. The relationship between pro-ADM and PCT and also APACHEⅡ score was analyzed with Pearson correlation analysis. The receiver-operating characteristic curve (ROC) of pro-ADM and PCT were used to evaluate the diagnostic acuity of sepsis.Results The plasma levels of pro-ADM, PCT and APACHEⅡ score in sepsis group were significantly higher than those in ordinary infection group and healthy control group [pro-ADM (ng/L): 66.69±1.73 vs. 53.43±2.70, 45.87±1.43; PCT (ng/L):1 336.49±40.26 vs. 1 083.09±47.99, 959.04±37.53; APACHEⅡ score: 14.60±0.81 vs. 8.10±1.14, 3.00±1.15,allP< 0.01]. With the aggravation of sepsis, the levels of pro-ADM, PCT and APACHEⅡ score were gradually increased, and there were significant differences among sepsis, severe sepsis, and septic shock groups [pro-ADM (ng/L): 64.91±2.50, 73.56±2.80, 84.67±4.52; PCT (ng/L): 1 152.65±48.62, 1 233.93±63.06, 1 475.71±109.93;APACHEⅡ score: 12.91±1.15, 14.55±1.14, 19.37±2.40,P< 0.05 orP< 0.01]. Pearson correlation analysis results showed that the level of pro-ADM was positively related with PCT (r = 0.473,P = 0.006), and it was also positively correlated with APACHEⅡ score (r = 0.707,P = 0.008). ROC curve analysis showed that area under the ROC curve (AUC) of pro-ADM for diagnosis of sepsis was 0.823 (P = 0.003). When the cutoff value was 59.40 ng/L, the sensitivity was 80.7%, the specificity was 68.0%, the positive predictive value was 85.2%, and the negative predictive value was 60.7%. AUC of the PCT for diagnosis of sepsis was 0.653 (P = 0.043). When the cutoff value was 1 194.67 ng/L, the sensitivity was 68.4%, the specificity was 64.0%, the positive predictive value was 81.8%, and the negative predictive value was 44.7%. It was proved that the pro-ADM had a higher diagnostic value for sepsis than PCT.Conclusion The plasma levels of pro-ADM can be used as an early indicator in diagnosis and severity evaluation and prognosis in patients with sepsis .

Animals ; Female ; Galactosylceramides ; metabolism ; Graft vs Host Disease ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes ; cytology ; Th2 Cells ; cytology

Objective To investigate the effect of astragalus polysaccharides (APS) on the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) to neurones, adipocytes, osteoblasts and chondrocytes, and provide basis for the development of effective and low toxic differentiation inducing agents. Methods BMSCs were isolated from SPF Wistar rats, purified, expanded and cultured to family 3. The appropriate concentration of APS was filtered out by MTT assay. The F3 cells were randomly divided into control group and induced group (neural induction, adipogenic induction, osteogenic induction, cartilage induction). The effects of APS and classical chemical drugs on differentiation were measured by toluidine blue, oil red o and alizarin red staining. The protein expression of NSE, LPL, collagen Ⅰ and collagen Ⅱ were examined by Western Blot. Results MTT assay showed that 1 g/L APS promoted the proliferation better than other concentrations especially in 48 hours. The morphologic change of the cell from BMSCs was uniformly positive to toluidine blue staining, and was negative to oil red o and alizarin red staining. Western blot showed that the protein expression of the cell from BMSCs was positive for NSE but negative for LPL, collagen Ⅰ and collagen Ⅱ. Conclusion BMSCs induced by APS can differentiate to neurone and fail to differentiate to adipocytes, osteoblasts and chondrocytes in vitro.

entrate graft can block pathologic process and improve the inclusion of femoral head and increase blood circulation of femoral head, which is beneficial to its recovery.

Objective To investigate the clinical roles of lysine methyltransferase 5A(KMT5A)in human hepatocellular carcinoma(HCC)and its functions in cell migration and invasion. Methods The expression levels of KMT5A of 60 cases were detected by immunohistochemistry(IHC). KMT5A siRNA was used to down-regulate the expression of KMT5A in SMMC-7721 cells. Cell migration and invasion were measured by wound healing assays and transwell assays,respectively. Immunoblotting was used to detect the expression of MMP-2 after siRNA trans-fection. miR-186 mimics were transfected into SMMC-7721 cells and the mRNA levels of KMT5A was detected by qRT-PCR after transfection. Results High expression of KMT5A was associated with large tumor diameter (>5 cm,P=0.047)and advanced TNM stage(Ⅲ+Ⅳ,P=0.035). The expression of KMT5A was knocked down by siRNA in SMMC-7721 cells. Down-regulation of KMT5A suppressed cell migration(P=0.031,P=0.006)and invasion(P=0.010),and impaired MMP-2 expression(P=0.040). Overexpression of miR-186 could significantly inhibit the expression of KMT5A(P = 0.007). Conclusions Over-expression of KMT5A in HCC tissues associ-ates with poor clinical features. KMT5A knockdown inhibits the migration and invasion on HCC cells.

OBJECTIVE: To discuss the application of E-commerce system in Hospital drug management. METHODS: It analyzed the defects existed in current Hospital Information System (HIS), introduced hospital E-commerce system and evaluated comparatively the drug management information before and after the application of the e-commerce system. RESULTS & CONCLUSIONS: E-commerce system has effectively made up the functional insufficiency of HIS and changed the traditional drug purchase mode; therefore, its application has a bright future.

Objective To investigate the level of psychological capital of nursing administrative personnel and analyze its influencing factors.Methods A total of 302 nursing administrative personnel of Anhui Province were investigated with Chinese Version of Nurses Psychological Capital Questionnaire (PCQ-24) and a self-designed question-naire about demographic information.Results The overall score of psychological capital was (106.23±10.67) points,and the dimension scores from high to low were selfefficacy (26.79±3.40) points,resilience (26.60±3.25) points,hope(26.56±3.55) points,and optimism(26.27±3.34) points.Multiple regression analysis showed that health?level and positions entered the regression equation,t=2.904,1.838,P＜0.05.Conclusions The overall nursing administrative personnel psychological capital is in the middle level,health level and positions were influencing factors of nurses psychological capital.It is suggested that the psychological capital should be introduced into nursing administrative field and take diversified measured to improve their psychological capital.

Objective To compare the diagnostic reliability of AO classification and Evans-Jensen classifications in X ray film and three-dimensional CT reconstruction images intertochanteric fractures, and explore advantage of the three-di?mensional CT. Methods A retrospective study was performed to evaluate 54 patients with intertochanteric fractures. Three orthopaedic surgeons were asked to make assessment of fracture classifications using X ray film and the three-dimen?sional CT images. Agreement test was performed to evaluate interobserver and intraobserver reliability for fracture classifica?tion. Results When X ray film was used, mean Kappa values of interobserver reliability for AO and Evans-Jensen classifi?cations between three surgeons were 0.597 and 0.571, that was medium consistency. While using three-dimensional CT to AO classification, lower Kappa value was 0.411, medium consistency. Evans-Jensen classification, Kappa value was 0.704, highly consistency. Each surgeon respectively using X ray film and three-dimensional CT, the mean Kappa values of AO and Evans-Jensen classification were 0.464, medium consistency and 0.191 (0.160-0.233), weak consistency. Conclusion Both classifications based on X ray film show moderate consistency, using three-dimensional CT does not improve the consis?tency of AO classification. The three-dimensional CT is not applicable in Evans-Jensen classification.