Objective To investigate the relationship between the changes of serum total bilirubin(TBil) and hybrid muscle crea-tine kinase isozyme(CK-MB),and its clinical implication in neonates with hypoxic-ischemic encephalopathy (HIE). Methods The levels of TBil and CK - MB were measured in 49 neonates with HIE (15 mild cases, 20 moderate cases and 14 serious cases), and 31 normal neonates on the first day and the fourth day after birth. Results The levels of TBil on the first day after birth were significantly higher than those in normal neonates;the levels of TBil were correlated with the severity of HIE(P

OBJECTIVEExpress and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.

METHODSThe expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.

RESULTSSuitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.

CONCLUSIONSWe have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.

DNA, Single-Stranded ; genetics ; metabolism ; DNA-Binding Proteins ; chemistry ; isolation & purification ; metabolism ; Hepacivirus ; Hepatitis C ; metabolism ; virology ; Humans ; Molecular Weight ; Protein Binding ; RNA, Viral ; genetics ; metabolism

Objective To investigate the expressions of indoleamine 2,3-dioxygenase (IDO) and transcription factor forkhead box P3(FOXP3)in gallbladder adenocarcinoma; and to evaluate the relationship between the expressions of IDO and FOXP3 protein,and clinicopathology and lymph node metastasis of gallbladder carcinoma.Methods The expressions of IDO and FOXP3 in 51 cases of primary gallbladder cancer,90 cases of chronic cholecystitis,and 20 cases of normal gallbladder tissues were studied by immunohistochemical staining. Results The positive expression rates of IDO and FOXP3 in gallbladder carcinoma were 72.5％ (37/51) and 60.8％ (31/51),in normal gallbladder mucosa were 5％ (1/20) and 20.0％ (4/20),in cholecystitis and gallstone were 11.1％ (10/90) and 32.2％ (29/90),respectively.There were significant differences among the three groups in IDO and FOXP3 expressions (P＜0.01).The positive expression rates of IDO and FOXP3 were 7.1％ and 14.3％ for simple hyperplasia,17.7％ and 35.3％ for atypical hyperplasia,40％ and 35％ for gallbladder carcinoma (stages Ⅰ-Ⅲ),and 77.4％ and 64.5％ for gallbladder carcinoma (stages Ⅳ-Ⅴ ).There were significant differences among the four groups in IDO and FOXP3 expressions (P＜0.01).There were significant differences among the Nevin stage groups,lymph node metastasis group and gallbladder stone in IDO and FOXP3 expressions.However,there were no significant differences among the sex groups and the age groups in IDO and FOXP3 expressions (P＞0.05).In gallbladder carcinoma,the expression of IDO had a positive correlation with FOXP3 expression in lymphocyte (γ=0.406,P=0.003).Conclusion In gallbladder cancer,a high-expression of IDO and an expression of FOXP3 in lymphocyte are closely related with immune tolerance of gallbladder carcinoma.The results provide a reference for clinical use of immunotherapy in gallbladder cancer.

Objective To assess the effect of suppression of insulin-like growth factor-1 receptor (IGF1R)in HO8910PM cell line by small interference RNA(siRNA).Methods Transfection of siRNA using liporectamine 2000 was conducted to silence IGF1R gene expression,the expression levels of IGF1R mRNA and protein were evaluated,and the effects on the cell cycles at 48 hours of transfection were assessed by real-time PCR,western blot and flow cytometry(FCM)assay respectively.The cell growth was detected by cell counting kit-8(CCK-8)at 24,48,72,96 hours of transfection.After 24 hours of transfection,the cells were cuhured with difierent concentrations of cisplatin(DDP)for 24 hours,the cell growth inhibition rate Was evaluated by CCK-8.Following incubation with 10μg/ml DDP for 24 hours after 24 hours of transfection,the apoptosis cells and the protein expression level of apoptosis-related gene,B cell leukemia/lymphoma 2(Bcl-2),were identified by FCM and western blot respectively.Resuits (1)Expression levels of IGF1R mRNA and protein were markedly decreased respectively at 48 hours of transfection IGF1R siRNA.(2)Suppression of IGF1R accompanied the reduction of cell growth at 48,72,96 hours of transfection with IGF1R siRNA,absorbance were 1.71±0.13,2.32±0.23,2.79±0.28 respectively (P＜0.01).(3)IGF1R siRNA induces arrest of G2 phase,the G2 phase rate of cells were 24.37%(P＜0.05).(4)Following treatment with 2.5,5,10,20μg/ml DDP for 24 hours after 24 hours of transfection,the cell growth inhibition rates were(25.94±0.08)%,(40.25±0.05)%,(59.48±0.03)%and(74.18±0.08)%respectively(P＜0.01).(5)Treatment with 10μg/ml DDP for 24 hours after 24 hours of transfection,induces 17.95%of cells apoptosis(P＜0.05),and decreases Bcl-2 protein level.Conclusion RNA interference of IGF1R gene induces the IGF1R silence in HO8910PM cell line significantly,inhibits cell growth in vitro, arrests the G2 phase, and enhances the chemosensitization to DDP.

Objective: To establish the quality standard for compound Heishen pills. Methods: Scrophulariae Radix, Radix et Rhizoma and Belamcancae Rhizoma were identified by TLC. HPLC was used to determine the content of harpagoside and cinnamic acid in Scrophulariae Radix on a Welchrom-C18 column using methanol-acetonitrile-1% ethylic acid (8∶21∶71) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 30℃ and the detection wavelength was set at 278 nm. Results:The TLC method had good specificity without interference from negative control. The linear range of harpagoside was 1. 32-65. 80μg·ml-1 with the average recovery of 98. 06%(RSD=2. 16%),and that of cinnamic acid was 0. 38-19. 20 μg·ml-1 with the average recovery of 98. 78%(RSD=1. 34%). RSDs of precision, stability and reproductibility tests were all below 2%. Conclusion: The established method is accurate, feasible and reproducible. It can be used in the quality control of compound Heishen pills.

[Objective]To investigate the significance of the high-risk human papilloma virus (HPV)detection in the screening and diagnosis of cervical lesions.[Methods] The high-risk HPV DNA test results of 797 patients with cervical lesions who all accepted cytology and histopathology test were collected and analyzed retrospectively.[Results]The high-risk HPV DNA positive rates in cervicitis,cervical intraepithelial neoplasia(CIN)Ⅰ,CIN Ⅱ,CIN Ⅲ and cervical cancer were 53.41％(188/352),70.91％(117/165),87.63％(85/97),97.90％(140/143),97.50％(39/40),respectively.The sensitivity and negative predictive value of the high-risk HPV DNA detection for CIN Ⅱ and more serious lesions were 96.66％(318/329),93.29％(153/164),respectively.The detection rate of CIN Ⅱ and more serious lesions in patients with atypical squamous cells of undetermined significance(ASCUS)and positive high-risk HPV DNA was 30.03％(94/313),while the rate in patients with negative high-risk HPV DNA was 1.55％(2/129).[Conclusions] The more serious the cervical lesion is,the higher high-risk HPV DNA positive rate is.It is most closely related with CIN 11 and cervical cancer.The high-risk HPV DNA detection has high sensitivity and negative predictive value for CIN Ⅱ and more serious lesions.The high-risk HPV DNA detection has high negative predictive value in CIN Ⅱ and more serious lesions in ASCUS.

BCG Vaccine ; adverse effects ; Humans ; Immunologic Deficiency Syndromes ; complications ; Panniculitis ; etiology ; T-Lymphocytes ; Tuberculosis ; etiology

Selective salpingography and transvaginal catheter dilatation using a self- made coaxial catheter set were performed in 45 women with infertility and fallopian tube ob- struction.The successful catheterization rate was 95.5% for fallopian tubes and the recanal- ization rate proximal allogian tubes was 80%,Normal intrauterine pregnancy was achieved in total 15 women(33.3%)with in two years follow-up 50% and 17.4% pregnant rates were obtained in secondary infertility and primary infertility,respectively.The authors be- lieve that transvaginal catheter dilization is a useful method for female infertility with fallopi- an tube obstruction,especially for the secondary infertility and proximal fallopian tube ob- struction.The self-made coaxial catheter set holds the advantages of effectiveness,safety, and inexpensiveness.

OBJECTIVETo explore the viral and host causes of hepatosteatosis in Chinese patient with chronic hepatitis B.

METHODSA total of 562 patients (450 males and 112 females, age range 13-80 years) with biopsy-proven chronic hepatitis B were enrolled in the study. The patients were divided into two groups: group without steatosis (460 patients) and group with steatosis (102 patients). The groups were compared in terms of age, gender, body mass index (BMI), liver enzymes, cholesterol, triglyceride, APO-A, APO-B, urine acid (UA), fasting serum glucose (FSG) and HBeAg, viral load.

RESULTSSteatosis was present in 102 patients (18.15%). The degree of liver steatosis in 97 (95.10%) patients were less 30%. Steatosis was found in 98 (21.78%) of male patients and 4 (3.75%) of female patients (P < 0.01). In the group of chronic hepatitis B with steatosis, the prevalence of obesity, diabetes mellitus, dyslipidaemia, alcoholic consumption, the BMI, cholesterol, triglyceride, UA and FSG levels were significantly higher than those in the group without steatosis (P < 0.01). No significant difference was found in the mean age, HBeAg, viral load between the two groups (P > 0.05). Logistic regression analysis demonstrated that the present of steatosis was positively correlated to BMI, TG and UA.

CONCLUSIONHepatosteatosis in chronic hepatitis B appears to be a result of metabolic factors of the host rather than the effect of viruses.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Fatty Liver ; complications ; genetics ; pathology ; virology ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; complications ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Prevalence

The purpose of this study is to explore depression metabolic markers in rat hippocampus and to investigate the anti-depressant effect of genipin and its mechanisms using nuclear magnetic resonance (NMR) metabonomics. Chronic unpredictable mild stress (CUMS) procedure was conducted to establish the depressive rat model. At the beginning of the third week, genipin low dose (25 mg x kg(-1)), middle dose (50 mg x kg(-1)), high dose (100 mg x kg(-1)), and venlafaxine (50 mg x kg(-1)) were given to the CUMS rats separately once daily for two weeks except control and model groups. Rat hippocampus was analyzed by 1H NMR based metabonomics after drug administration for 2 weeks. Significant differences in the metabolic profile of rat hippocampus of the CUMS treated group and the control group were observed with metabolic effects of CUMS including decreasing in glycine and N-acetylaspartate, increasing in inositol, glutamate, lactate, glutamine, taurine and alanine. Genipin showed ideal antidepressive effects at a dose of 50 mg x kg(-1) in rats, decrease of inositol, glutamate, lactate, alanine were observed, while glycine and N-acetylaspartate were increased. Important influence has been found on normal nervous system function of these significant changed metabolites, which suggests that the antidepressant effect of genipin may be played by enhancing the activity of neurons in hippocampus, repairing and improving the function of the neuron. The metabonomics approach is an effective tool for the investigation of the anti-depressant effect and pharmacologic mechanisms of genipin.
Alanine ; metabolism ; Animals ; Antidepressive Agents ; isolation & purification ; pharmacology ; Aspartic Acid ; analogs & derivatives ; metabolism ; Behavior, Animal ; drug effects ; Chronic Disease ; Depression ; drug therapy ; metabolism ; physiopathology ; Gardenia ; chemistry ; Glutamic Acid ; metabolism ; Glycine ; metabolism ; Hippocampus ; drug effects ; metabolism ; Inositol ; metabolism ; Iridoids ; isolation & purification ; pharmacology ; Lactic Acid ; metabolism ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley