In this study, plate transparent circle by Davis method was introduce firstly screening ?-Rhamnosidase high-yield strain. The spore-sprouted Aspergillus niger 8-hour were mutagenized by ethyl methane sulphonate and pre-screened via transparent circle. 11% mutants yield 40% higher of ?-rhamnosidase than the original strain. A high-yield strain, T-226 with the highest ?-rhamnosidase activity of 373.4 U/mL was finally selected from these potential high-yield mutants after rescreened by shake flask fermentation twice. When the T-226 strain was fermented in 5 L bioreactor, the enzyme activity could reach to 631.9 U/mL after 84 h. Thus, the established screening method is highly efficient to isolate ?-rhamnosidase high-yield mutant of A. niger.

Objective To explore the MRI finding of female normal levator ani muscle and the levator ani muscle injuries and to evaluate the correlation between childbirth and levator ani muscle injuries. Methods One hundred asymptomatic nulliparous women(control group)and 200 vaginally primiparous women(study group)were selected as the object of this study.Moreover,the study group was divided into two subgroups:group A(100 cases)with stress incontinence,group B(100 cases)without clinical symptoms.Multiplanar proton density magnetic resonance images were obtained at 0.5 cm intervals from these study individuals.All images were used to analyze the differentiation of MRI features between normal levator ani muscle and levator ani muscle injuries.Results No levator ani injuries were identified in the control group.Fifty-four primiparous women(27%)had visible injuries in their levator ani muscles,42 in group A and 12 in group B.Injuries were identified in the puborectalis muscle in 49 cases and in the iliococeygeus muscle in 5 cases(X~2=41.447,P

Objective To induce cord blood derived endothelial progenitor cells(EPCs)into endothelial cells,and investigate the feasibility of these cells as the seed cells of tissue engineering cardiovascular replacement.Methods Mononuclear cells were isolated from fresh cord blood by 6%HES and density gradient centrifugation.Isolated cells were cultured in medium supplemented with vascular endothelial growth factor(VEGF).Attached cells were identied by morphology,immunofluorescence staining and flow cytometry.Results The percentage of mononuclear cells isolated from fresh cord blood was(3.4?2.1)?10~7/mL.The morphology of attached cells changed while being cultured and inducted,from small-sized round cells to spindle-like cells,to a typical cobblestone morphology,and the total number of cells was 10~7.After 7 days of culture,immunofluorescence staining showed that the vWF and VEGFR-2 were expressed.Compared with the original,cell markers CD_133 decreased(3.11%?1.05%) to(0.09%?0.02%),P

Objective To investigate the effects of different duration hypotension thresholds on p-Tau-181 and Aβ-42 protein expression and cognition in rats. Methods Thirty-nine healthy male SD rats were randomly di-vided into 4 groups:the control group(group C,n=9),the hypotension group(groupA1、A2、A3 ,n=10). The blood pressure of groupA1、A2、A3 was measured in different time of 2 h、4 h、6 h ,for 5 days. The antihyperten-sive group of mean arterial pressure(MAP)were maintained in the 50~55 mmHg safe range. Morris water maze was used to detect the spatial learning and memory ability of rats. The levels of Aβ42 and p-Tau-181 were detected by ELISA. Results There was no significant difference in mortality of rats in each group (P > 0.05). Compared with the group C,the escape latency and swimming distance of A2 group and A3 group were increased(P<0.05). In 3~7 days after operation,the cerebrospinal fluid P-Tau-181 and Aβ42 protein expression increased in the A2 group and A3 group compared with the A1 group(P<0.05). The escape latency and swimming distance of the A2 group and the A3 group were significantly longer than those in the control group. Aβ42 and p-Tau-181 were signifi-cantly increased in A3 group(P < 0.05). Compared with the A2 group,the increase of Aβ42 and p-Tau-181 in the A3 group was not significant(P<0.05). Conclusion Long-term controlled hypotension may lead to postoper-ative cognitive dysfunction which may relate to the increase of Aβ42 and p-Tau-181 protein expression.

OBJECTIVETo investigate the clinical effects and prevent the complications of posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure.

METHODSFrom 2008 January to 2011 December, 17 patients with cervical anterior internal fixation failure were treated with posterior and anterior decompression and internal fixation. There were 12 males and 5 females, aged from 26 to 68 years old with an average of 44.1 years. The lower screw loosening was found in 6 cases, the upper screw loosening in 5 cases, titanium mesh caving in 3 cases, the upper screw breakage in 2 cases, the lower screw breakage in 1 case. Informations of bone fusion were observed by X-ray, CT, MRI. Clinical effects were evaluated by modified JOA score.

RESULTSAll the revision operations were successfully completed. One case with poor blood coagulation function before operation resulted in postoperative hematoma and occurred neurological symptoms; after hematoma removal and fresh frozen plasma infusion later, neurological symptoms of the patient disappeared. All patients were followed up from 6 to 38 months with an average of (22.4±10.0) months. Postoperative at 2 weeks, 3 months, and final follow-up, JOA score had obviously improved and respectively was 13.1±1.6, 13.4±1.6, 14.2±1.5. All internal fixation locations were good after revision,and obtained bone fusion at 10 months after operation, with an average fusion time of 6 months.

CONCLUSIONThe combined posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure is safe, can achieve thoroughly decompression, maintain the cervical curvature, reconstruct the three column stability, and it may be used for the patients of cervical anterior fixation failure.

Adult ; Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

OBJECTIVETo explore the clinical value of CT-guided percutaneous needle biopsy (PCNB) for diagnosis of atypical pulmonary tuberculosis.

METHODSThe data from 15 patients undergoing diagnostic PCNB for pulmonary tuberculosis were retrospectively analyzed. Prior to PCNB, definitive diagnosis failed to be obtained from other examinations including sputum smear in 10 cases, fiberoptic bronchoscopy in 12, chest X-ray in 13, CT scan in 15, and position emission tomography (PET) with CT examination in 1 case.

RESULTSSputum smears and fiberoptic bronchoscopy both failed to produce positive results and pulmonary and/or mediastinal lesions were shown by chest X-ray, CT, and PET-CT. Pulmonary tuberculosis were diagnosed in the 15 cases by histopathological examination following PCNB, and pneumothorax in relation to PCNB occurred in 3 cases and hemoptysis in 1 case.

CONCLUSIONPCNB provides a safe and accurate means for diagnosis of pulmonary tuberculosis when imaging and other diagnostic modalities fail to result.

Adolescent ; Adult ; Biopsy, Needle ; methods ; Female ; Humans ; Male ; Middle Aged ; Radiography, Interventional ; Reproducibility of Results ; Retrospective Studies ; Sensitivity and Specificity ; Tomography, X-Ray Computed ; Tuberculosis, Pulmonary ; diagnosis ; diagnostic imaging ; pathology

OBJECTIVETo study the effects of Rapamycin (RPM) on angiogenesis and tumor progression in human hepatocellular carcinoma (HCC) implantation mice.

METHODSTumor tissues of HCC were implanted into the liver of nude mice. Then, nude mice were treated with RPM and cyclosporine A (CsA). Real-time PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF). Immunohistochemical stain and image analysis were used to detect the protein expression of VEGF and proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) was counted by endothelial cells immunostained by anti-CD34 antibody. The concentration of VEGF in the peripheral blood was detected by ELISA.

RESULTS(1) The tumor weights were (372 +/- 35) mg, (769 +/- 39) mg and (751 +/- 42) mg in RPM, CsA and control group respectively. The tumor weight was significantly decreased in RPM group and no difference in CsA group compared with control group. (2) The expression of VEGF mRNA, VEGF and PCNA protein in tumor tissues and concentration of VEGF in the peripheral blood were remarkably down-regulated in RPM group compared with control group (P < 0.05) and were not remarkably different in CsA group from in control (P > 0.05).(3) Comparing with the control, the tumor MVD was remarkably decreased in RPM group (P < 0.05), and no difference in CsA group (P > 0.05).

CONCLUSIONRPM can inhibit angiogenesis and tumor progression of HCC by down-regulated the gene and protein expression of VEGF and inhibited the growth of tumor.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Nude ; Neovascularization, Pathologic ; drug therapy ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; genetics ; Sirolimus ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Xenograft Model Antitumor Assays

BACKGROUNDMostly because of the limited number and proliferative ability of the transplanted hepatocytes, hepatocyte transplantation offers only temporary support to the hepatic function with rather poor functional replacement of the damaged liver parenchyma. This study aimed to observe the therapeutic effect of human thioredoxin (hTrx) gene-modified hepatocytes on experimental acute liver failure in rats.

METHODShTrx cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma 143 (TK-) cells to construct the recombinant retrovirus vector pLEGFP/hTrx, which was packaged into PA317 cells to collect the recombinant retrovirus containing hTrx gene. After titration and characterization, the recombinant retrovirus was applied to primary cultured rat hepatocyte for infection to generate hTrx gene-modified rat hepatocytes, whose viability and antioxidative capacity were examined by immunohistochemistry and MTT assay, respectively. In a Sprague-Dawley (SD) rat model of acute liver failure, the modified hepatocytes were injected into the spleen, and the hepatic function and survival rate of the recipient rats were evaluated at different time points after the transplantation.

RESULTSNIH3T3 cells infected by the recombinant retrovirus were capable of expressing bioactive hTrx in the form of fusion proteins. Immunohistochemistry demonstrated normal function of the hTrx gene-modified hepatocytes, which possessed strong antioxidative capacity as shown by MTT assay. Transplantation of the modified hepatocytes in rats with acute liver failure resulted in significantly lowered serum alanine aminotransferase (ALT) and total bilirubin (TBIL) levels (P < 0.05). The hepatocytes exhibited long-term survival and efficient proliferation after transplantation. Fourteen days after the operation, the rat models receiving hTrx gene-modified hepatocytes had significantly higher survival rate than those without the transplantation.

CONCLUSIONhTrx gene-modified hepatocyte transplantation can effectively alleviate acute liver failure in rats.

Animals ; Hepatocytes ; metabolism ; transplantation ; Humans ; Liver Failure, Acute ; therapy ; Mice ; NIH 3T3 Cells ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Thioredoxins ; genetics

OBJECTIVETo study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice.

METHODSHepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay.

RESULTSThe tumor weight was (352+/-38) mg, (683+/-53) mg and (675+/-45) mg in SRL, FK506 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.01), and there was no difference between FK506 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01).

CONCLUSIONSRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.

Animals ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Immunohistochemistry ; Liver ; blood supply ; pathology ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Proliferating Cell Nuclear Antigen ; metabolism ; Sirolimus ; administration & dosage ; pharmacology ; Tacrolimus ; administration & dosage ; pharmacology ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

Objective To estimate the prevalence of overweight and obesity , and their relationship between overweight , obesity and chronic diseases among adults in Pinghu City . Methods A total of 3 106 local registered permanent residents aged above 18 years were selected by multi-stage random sam-pling and questionnaire surveys were conducted .The blood glucose , blood lipid , blood pressure , height and weight of the participants were tested . Results The prevalence rate of overweight was 31 .6%( the standardized rate was 29 .5%) and the prevalence rate of obesity was 8 .1% ( the standardized rate is 7 .8%) in residents aged above 18 years in Pinghu City .The prevalence rate of overweight and obesity was highest among population aged from 50 to 59 years.The prevalence rates of overweight of the population in different ages and with different education backgrounds had significant differences ( P <0.05 ).The prevalence rates of diabetes , hypertension and dyslipidemia among overweight and obese population were higher than those with normal weight ( P <0 .01 ) .BMI was the risk factor of hypertension , diabetes mellitus and dyslipidemia , and the OR value was 1 .508 , 2 .127 and 1 .571 , respectively . Conclusion The prevalence of overweight and obesity in Pinghu City is serious and has close relation with chronic diseases .Prevention and intervention measures are necessary for the overweight and obesity population .