Objective To evaluate the inhibitory effect of the small interfering RNA(siRNA) on hepatitis B virus(HBV)in vivo which targets HBV S gene region.Methods An animal model of HBV infection was developed hydrodynamically by injecting pcDNA3.1-HBV together with siRNA through the tail vein of Balb/c.HBsAg was analyzed by time resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR(FQ-PCR),HBV S-mRNA was detected by semi-quantitative RT-PCR,and viral specific proteins(HBsAg and HBcAg)in the liver were assayed by immunohistochemical staining.Results In the mice,the siRNA could effectively inhibit the secre- tion of HBsAg,reduce the titers of HBV DNA,and immunohistochemical results also indicated that the number of HBsAg and HBcAg positive cells was reduced.The inhibitory effect of siRNA on HBV lasted 3 clays at least.Conclusion These results demonstrate that the siRNA targeting HBV S gene region can substantially and specifically inhibit HBV replication and expression in vivo.

Objective To explore the activation of the primary somatosensory cortex (S_Ⅰ) and secondary somatosensory cortex (S_Ⅱ) undermild and moderate pain. Methods 7 healthy right-handed volunteers accepted electrostimulation of 1 and 2 times pain threshold respectively.The severity of pain was determined with the Visual Analogue Score (VAS), and they received functional magnetic resonance imaging(fMRI) simultaneously. Results 1 and 2 times pain threshold electrical stimulation caused mild and moderate pain respectively. Bothmild and moderate pain activated contralateral S_Ⅰ similarly, and bilateral S_Ⅱ with pain intensity. Conclusion S_Ⅰ and S_Ⅱ respond to the mildand moderate pain in different ways.

A total of 1151 subjects were enrolled in this study.Metabolic syndrome (MS) was diagnosed according to the International Diabetes Federation (IDF) criteria.Significant differences in waist circumference,body mass index(BMI),diastolic blood pressure(DBP),systolic blood pressure(SBP),fat mass,Fat％ in different serum TSH levels were found.There were positive relation between fasting plasma glucose,DBP,SBP,and serum FT4 levels,between high density lipoprotein-cholesterol,DBP,SBP,waist circumference,fat mass,Fat％,and serum FT3 levels,even after adjustment for age and sex.Serum FT3 and FT4 levels were higher in the MS group than those in the control group.

Objective To evaluate the safety and efficacy of magnetic resonance-guided focused ultrasound surgery (MRgFUS) in treatment of symptomatic uterine leiomyoma among Chinese reproductive age women.Methods From April 2010 to April 2012,80 premenopausal women with symptomatic leiomyoma volunteered to participate in this prospective study in Department of Outpatient of Peking Union Medical College Hospital.Among 23 reproductive aged patients with size of uterus less than 16th gestational weeks,2.5 to 10 cm of diameter of myoma,less than 10 myomas and expressing symptoms clearly were treated by MRgFUS.Treatment data,non-perfused volume ratio (NPVR) and adverse events were recorded.After treatment,patients were followed up at 1 week,1,3,6,12 and 24 months,respectively.Patients at initial screening and each time of the follow-up filled out uterine fibroid symptoms quality of life (UFS-QOL),which include symptoms severity score (SSS) and health-related quality of life (HRQL).The volumes of leiomyoma and uterine were evaluated on MRI before and after the treatment (at 6 and 12 months,respectively).Before operation,routine blood test were performed on all patients,anemia patients at 3 months and 1 year after treatment were checked with blood test.Results (1) Treatment data and adverse events:the mean therapeutic temperature was (69 ± 7) ℃,the mean treatment time was (144 ±62) min,the mean NPVR was (62 ±23)％.Adverse events included mild erythema(1/23),abdominal cramp(8/23),vaginal discharge (5/23),and leg numbness (4/23).(2) The rate of secondary surgery:one patient was treated by myoectomy and hysterectomy within one year following up and 4 patients chose surgical treatment during the second-year follow-up.(3) Volume change:the volumes of leiomyoma before the treatment and 6,12 months after the treatment are 75.6 (P25 =43.8,P75 =128.9),52.3 (P25 =23.8,P75 =111.2),45.9 (P25 =26.3,P75 =71.7) cm3,respectively; and the volumes of uterine before the treatment and 6,12 months after the treatment are 270.0 (P25 =208.4,P75 =390.3),216.4 (P25 =151.1,P75 =290.0),200.0 (P25 =149.1,P75 =267.6) cm3,respectively.Both leiomyoma and uterine volumes decreased significantly after treatments (P ＜ 0.01).(4) UFS-QOL change:the symptoms severity score (SSS) before the treatment and 3,12 months after the treatment are (34 13),(22 ± 11),(19 ± 12),which decreased significantly (P ＜ 0.01).The health-related quality of life (HRQL) before the treatment and 3,12 months after the treatment are (74 ± 15),(82 ± 13),(89 ± 10),which increased dramatically (P ＜0.01).(5) Hemoglobin(HGB) change:eleven patients suffered from anemia before treatments,the mean HGB before treatment was (87 ±6) g/L and were (106 ± 14) g/L 3 months after treatment,(112 ± 10) g/L 12 months after treatment.The HGB was increased significantly after treatments (P＜0.01).Conclusions MRgFUS is a safe and effective non-invasive management for symptomatic uterine leiomyoma in short-term follow up.But there is additional treatment ratio after MRgFUS.

AIM:To investigate rat Urotensin-II(ra t U-II)-induced vasoconstriction of rat main pulmonary arteries and the role of mitogen-activated protein kinase(MAPK). METHODS: The main pulmon ary artery was dissected from the male Sprague -Dawley rats and artery ring width was 3-4 mm. Concentration-response curves wer e gene rated to rat U-II(0 03 nmol/L-30 nmol/L).Inhibitor of MAPK,PD 98059(0 1 ?mol/ L -10 ?mol/L) were added into the medium after rat U-II(30 nmol/L)induced vasoc onstriction had reached plateau to construct the relaxant concentration-respons e curves and their EC 50 and E max . RESULTS: Rat U-II was a potent vasoconstrictor of isolated rat main pulmonary arteries [EC 50 =7 95?0 4 0, E max =(14 28?6 34)% of the response to 60 mmol/L KCl]; PD 98059 caused c oncentration-dependent relaxations of rat U-II precontracted arteries [EC 50 =5 91?0 45, E max =(81 39?13 65)%]. CONCLUSION: Rat U- II was a potent vasoconstrictor of rat main pulmonary arteries and this response was med iated through MAPK.

OBJECTIVE: To investigate whether the in vitro sensitivity of cefazolin sodium to the bacteria was altered after nanoparticlization. METHODS: The minimal inhibitory concentrations (MIC) of cefazolin sodium before and after nanoparticlization to S. aureaus and E. coli. were determined by microdilution. RESULTS: The MIC of nanoparticlized cefazolin sodium to S. aureaus and E. coli. had no significant change compared with that of non-nanoparticlized one. CONCLUSION Nanoparticlization will not decrease the sensitivity of cefazolin sodium to the bacteria.
Anti-Bacterial Agents ; pharmacology ; Cefazolin ; pharmacology ; Escherichia coli ; drug effects ; Microbial Sensitivity Tests ; Nanostructures ; Staphylococcus aureus ; drug effects

OBJECTIVE: To explore the effect of suppressive oligodeoxynucleotides (Sup ODN) on the Th1 differentiation of CD4(+)T splenetic lymphocytes in mice. METHODS: The splenetic lymphocytes of BALB/c mice were separated, and then CD4(+) cells were purified with immune magnetic CD4(+) microbeads (positive selection). The purification was examined by fluorescence-activated cell sorter. CD4(+) cells, anti-CD3epsilon, anti-CD28, IL-12 and Sup ODN or control oligodeoxynucleotides (Con ODN) were co-incubated for 72 h. IFN-gamma and IL-4 in the supernatant were detected using enzyme-linked immunosorbent assay. The expression of T-bet mRNA in CD4(+) cells was tested by reverse transcription polymerase chain reaction. RESULTS: Sup ODN could significantly inhibit the release of INF-gamma and increase IL-4 production respectively (P<0.01). T-bet mRNA of CD4(+) lymphocytes was remarkably inhibited by Sup ODN as well (P<0.01). CONCLUSION In the presence of pro-Th1-cytokines, Sup ODN may affect the differentiation of CD4(+) T lymphocytes in vitro. Sup ODN can promote CD4(+) T cells to differentiate into Th2, and suppress them into Th1.
Animals ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; Cell Differentiation ; drug effects ; Female ; Interferon-gamma ; antagonists & inhibitors ; physiology ; Interleukin-12 ; antagonists & inhibitors ; physiology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides ; pharmacology ; T-Lymphocyte Subsets ; Th1 Cells ; cytology ; immunology

OBJECTIVETo establish a new method for the determination of cyclovirobuxine-D in huangyangning tablets.

METHODAgilent Zorbax Extend C18 (4.6 mm x 250 mm, 5 microm) column with temperature 40 degrees C was used. The mobile phase was 0.3% diethylamine methanol solution and 0.3% diethylamine water solution, with a linear gradient of 0.3% diethylamine methanol solution from 78% to 95% in 9 minutes, and then maintained for 6 minutes. The flow rate was 0.8 mL x min(-1). Evaporative light scattering detector was used.

RESULTThe calibration curve was linear at a range of 0.57-11.44 microg for the cyclovirobuxine D (r = 0.9996). The average recovery was 97.2% and RSD was 1.1% (n = 9).

CONCLUSIONThis method is rapid, simple, and reproducible, and can be used as a quality control method for this preparation.

Buxus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Tablets

OBJECTIVETo explore the dynamic changes of fetuin-A expression and the influences of the changes on liver damage, hepatocyte apoptosis and inflammation in a mouse fulminant hepatic failure (FHF) model.

METHODSThe changes of fetuin-A expression were investigated by semi-quantitative RT-PCR and Western blot. Immunohistochemical staining was used in TNFa and fetuin-A detection. Hepatocyte apoptosis was detected by TUNEL.

RESULTSFetuin-A mRNA expression decreased after the FHF model was established for 3 hours (compared with the normal group, P less than 0.01), while the protein expression decreased after nine hours (compared with the normal group, P less than 0.01). Fetuin-A expressions were negatively correlated with the liver pathological scores and TNFa levels.

CONCLUSIONIn our mouse FHF model, fetuin-A is a possible protective factor for liver damage.

Animals ; Blood Proteins ; metabolism ; Female ; Liver ; metabolism ; pathology ; Liver Failure ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; alpha-2-HS-Glycoprotein

OBJECTIVETo observe whether H. pylori inoculated by oral route could arrive in livers and cause liver inflammation as an independent etiological factor.

METHODSC57BL/6 mice were orally inoculated with H. pylori SS1 strains and fed for 8 months. H. pylori colonization and pathologic consequences were studied in the liver and gallbladder tissues of the mice; the blood, liver tissue and gastric mucosa were obtained and cultured for H. pylori growth; The bacterial DNA extracted from the liver, bile and blood was examined by nested PCR for H. pylori genes. 16S rRNA PCR amplicons were sequenced and compared with the sequencing results of 16S rRNA PCR amplicons of the bacteria cultured from gastric mucosa and the inoculated H. pylori SS1.

RESULTSThe bacterial DNA extracted from the liver, bile and blood of the infected mice was detected for H. pylori genes by nested PCR. Six of the 15 samples were positive (40%) in the liver, 6 of 10 samples in the bile (60%), and 2 of 10 samples in the blood (20%). Sequencing results of 16S rRNA PCR products of the livers showed 100% homogeneity when compared with the cultured H. pylori from gastric mucosa and inoculated H. pylori SS1. H. pylori was found in 4 liver tissues of the 15 infected mice (26.7%) and 6 in the gallbladders (40%). Infiltrations of lymphocyte cells along hepatic sinusoids and a lower degree infiltration around interlobular arteries and veins were observed; ballooning degeneration was also observed in some hepatocytes.

CONCLUSIONH. pylori inoculated by oral route could arrive in the liver and cause inflammation as an independent etiological factor. The routes which the microorganisms took to reach the livers may involve hematogenous and/or biliary system dissemination.

Animals ; Helicobacter Infections ; Helicobacter pylori ; pathogenicity ; Liver ; microbiology ; Male ; Mice ; Mice, Inbred C57BL ; Rats