Objective To study the inhibition of HCV IRES mediated HCV core protein expression in cells by inhibitor RNA. Methods Plasmid pcRz-IRNA, a eukaryotic expression vector with IRNA and two self cleavage ribozyme overhang at both sides respectively, was constructed and co-transfected with pcHCVcluc (containing HCV NCR, core and Luc genome) into the HHCC cell line (Human Hepatocellular Carcinoma cell line). Immunoflurescence tests were applied to detect the co-transfected cells, which were thereafter analysed with confocal microscope quantitatively. Luciferase activity was valued using Luc Assay System (Promega). Results The cotransfected cells expressed HCV core protein, and the fluorecein in which was reduced significantly in comparison with control. Conclusions IRNA can inhibit the expression of HCV IRES mediated core protein in the cotransfected cells.

Objective To discuss the microsurgery approaches and the prevention and treatment of complication about the tumors in cerebellopontine angle.Methods The 53 cases with cerebellopontine angle brain tumor were retrospectively analyzed.Results In all 53 patients,the tumor of 23 cases were full-resected,29 cases' tumor were resected more then 3/4,only one cases' tumor was partly treated.Following operation,one case died,5 cases had hearing decreasing,3 cases had severely facial nerve weakness and non hearing,10 cases had facial nerve anaesthesia,futhermore there was one case with long-period coma because of the injury of brain stem.Conclusion The important factors that decrease the complications of cerebellopontine angle tumor surgery include surgery approachs,operating experience and mechanism of complication.

Objective To investigate expressions of programmed cell death ligand 1 (PD-L1) in hepatic tissues at the different stages of hepatitis B virus ( HBV) infection, and clarify its role in the mechanism of chronic hepatitis B virus infection. Methods The expressions of PD-L1 were detected by immunohistochemistry and computer image quantitative analysis in the hepatic tissues of 65 chronic HBV infected patients and 5 healthy controls. The correlations between PD-L1 expression and inflammatory grading in the hepatic tissues, total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum HBV DNA level were analyzed. Results The PD-L1 expressions in hepatic tissues of HBV infection with G0 - G4 inflammatory grades were 3. 07 % ±0.93%, 8.01%±1.49%, 11.60%±2.60%, 18.41%±2.21% and 26. 04% ±3. 41%, respectively,which were all significantly stronger than that in controls (0. 64%±0. 28%). PD-L1 expression was a positively correlated with inflammation grading of hepatitis tissues, TBil, ALT and AST level in serum (r=0. 917, 0. 787, 0. 483, 0. 628; all P<0. 05), and negatively correlated with serum HBV DNA load (r=-0. 620, P<0. 05). Conclusion The upregulated PD-L1 expression may be probably involved in the chronicity of HBV infection.

Objective To investigate the in vitro effects of compound Chinese herbal medicine Zilongjin on prostate cancer cell lines. Methods MTT assay,flow cytometry, colony on soft agar,and fluorescence microscopy were used to observe the effects of Zilongjin on anti proliferation,inhibition of clonal growth,cell cycle arrest and apoptosis induction.RT PCR method was used to study the regulative effect of Zilongjin on prostate marker genes PSA,AR,apoptosis related genes Bcl 2,Bax and tumor suppressing gene p16. Results Zilongjin caused notable anti proliferative effect,and induced G 0/G 1 phase arrest dose dependently on LNCaP,DU 145 and PC 3 cell lines.Inhibition of cell growth by 50% (IC 50 ) was observed with Zilongjin at concentrations of 0.79,0.42 and 0.52 mg/ml,respectively.Apoptosis induction was seen in LNCaP and DU 145 cell lines.Zilongjin also caused inhibition of clonal growth of DU 145 cell line.In addition,Zilongjin down regulated the expression of PSA,AR and Bcl 2 genes in LNCaP cell line;and down regulated Bcl 2,up regulated Bax and p16 genes expression in DU 145 cell line. Conclusions It is demonstrated that Zilongjin has antitumor action through anti proliferation,inhibition of clonal growth,G 0/G 1 phase arrest,apoptosis induction and regulating the expression of PSA,AR,Bcl 2,Bax and p16 genes.

[ABSTRACT]OBJECTIVETo study of the relationship of age and climate with the nasal mucosa lesions.METHODSAll the patients met with the inclusive criteria were registered and followed-up once a month and their nasal mucosal lesions were observed through nasal speculum and/or nasal endoscope. The recording data was analyzed by SPSS17.0 software.RESULTSA total of 259 patients, 555 person-times and 957 lesion spots were observed. The nasal mucosa was injured by airflow with the highest incidence rate from October to next year April. Patients numbers gradually increased after August, and significantly increased at November, December (P<0.05). Children aged from 2 to 12 years old and the adults aged from 40 to 80 years old were susceptible population (P<0.05), the incidence rate had a negative relation with humidity (P<0.05). The susceptible spots of the lesion were ranked as follows: Little area, inferior turbinate, mucosa of the both side of the deviated nasal septum, inferior meatus, middle meatus, middle turbinate and olfactory area. CONCLUSIONNasal mucosal injury relates to age and seasons. The nasal cavity airflow may be a factor of these lesions.

Objective To identify the relationship between viral factors and disease progression in patients with acute hepatitis B virus (HBV) infection. Methods Ninety-seven adult patients with acute HBV infection in Shanghai Changhai Hospital were enrolled in this study and followed up for 24 weeks. Epidemiological, biochemical and virological parameters of all patients were collected. HBV S region from sera of 54 patients with acute HBV infection were genotyped using direct nucleotide sequencing. Differences of means between groups were compared by t-test, and frequency between groups was compared by X test. Results The clinical manifestations of all patients were mild and the 83 patients spontaneously developed HBeAg and HBsAg seroconversion. However, 14 patients had a tendency of chronicity, with HBV DNA level higher than patients without chronicity tendency [(6. 17 ±1. 04) 1g copy/mL vs (3. 86±1. 85)1g copy/mL;t = 5. 95, P<0. 01]. Among the 14 patients, 6 obtained HBsAg seroconversion after antiviral therapy and the other 8 developed to be sustained HBV carrier who had not received antiviral therapy. The main genotypes of acute HBV infection were genotypes B and C. There were no statistically significant differences of epidemiological factors and biochemical results between patients with the two genotypes of HBV infection. High viral load at baseline was the risk factor of chronicity tendency. Conclusions The main genotypes of acute HBVinfection in Changhai Hospital in the year from 2003 to 2007 are genotypes B and C. There is no significant relationship between genotype and clinical outcome. While high viral load at baseline is significantly associated with chronicity tendency. Proper antiviral therapy can decrease sustained HBV infection rate.

ObjectiveTo investigate the effects of the imbalance between regulatory T cells (Treg) and T helper 17 cells (Th17) in patients with chronic hepatitis B virus (HBV) infection.MethodsThe serum concentration of Treg/Th17 differentiation-related cytokines in 34 patients with chronic hepatitis B (CHB),20 patients with HBV related acute on chronic liver failure (ACHBLF),and 20 healthy controls (NC) were measured by enzyme-linked immunosorbent assay (ELISA) and proportion of peripheral Th17 and Treg cells were analyzed by flow cytometry.Numeration data was analyzed by Fisher's exact propability method and measurement data was tested by one-factor analysis of variance or Turkey multiple comparison.Results The levels of Th17 differentiation-related cytokines,II-1β (3.97±2.85) pg/mL,IL-6 (12.75±-8.87) pg/mL,and IL-21 (360.0±335.7) pg/ mL in patients with ACHBLF were significantly increased than those in NC,which were (1.87 ±0.94) pg/mL(q=4.559,P＜0.01),(5.28±0.72) pg/mL(q=7.309,P＜0.01) and (46.68±20.17) pg/mL(q=6.946,P＜0.01 ),respectively.The proportion of Th17 increased markedly in patients with ACHBLF than that in NC(q=3.972,P＜0.05).However,compared to NC and patients with ACHBLF,the Treg differentiation-related cytokine,TGF-β,in patients with CHB,increased significantly (q=4.536 and 5.323,respectively; both P＜0.01).And the population of Treg also increased markedly in CHB patients.The level of IL-17A which was the characteristic effector cytokine of Th17 was the highest in patients with ACHBLF.The peripheral Th17 cell proportion was positively correlated with the level of serum total bilirubin in patients with ACHBLF (γ=0.74,P＜0.01).Conclusions Th17 and Treg imbalance including cytokine profiles and cell numbers exists in patients with chronic HBV infection.The Th17 are active in patients with ACHBLF and Treg are active in patients with CHB.

Objective To investigate the distribution of genotypes in chronic HBV infection (CHB) and acute HBV infection (AHB) patients in Shanghai. Methods Sixty-two patients with AHB and 73 patients with CHB admitted to ('hanghai Hospital of Shanghai between 2003 and 2007 were studied. Viral genotypes of all the patients were determined by direct gene sequencing.Meanwhile, epidemiological, clinical and biochemical parameters of all patients were collected. Mean values of different groups were compared by t test while frequency was compared by chi square test. Results The major prevalent genotypes in both AHB and CHB patients were genotype B and C (48.4% vs 51.6% in AHB patients and 26.0% vs 74.0% in CHB patients). The proportion of genotype B was higher in AHB patients compared to CHB patients (P= 0.02). Epidemiological factors and clinical outcomes were not statistically different among patients with different viral genotypes. The proportion of genotype C was much higher in CHB patients compared to AHB patients (P=0.006). The main transmission route of AHB was heterosexual interaction which was 18 out of 62 (29.0%), but in CHB patients, it was prenatal transmission which was 38 out of 73 (52.1%). Conclusions In shanghai, the main HBV genotypes in both AHB and CHB patients are genotype B and C. The proportion of genotype B is relatively high in AHB patients while proportion of genotype C is more common in CHB patients. There is no significant relationship between genotypes and the clinical outcomes of AI-IB patients.

Objective To evaluate the osteogenic characteristics of an injectable silk fibroin (SF) enhanced calcium phosphate cement (CPC) composite loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2) on lumbar interbody fusion in sheep. Methods Twenty-four mature sheep were randomly divided into two groups. Each sheep underwent L1.2, L3.4 and L5.6 lumber interbody fusion, and the three disc spaces were randomly implanted with three of the following materials: SF/CPC, CPC/rhBMP-2, SF/CPC/rhBMP2 and autogenous iliac crest bone. One group was killed at 6 months and the other at 12 months. The fusion segments were observed and analyzed by manual palpation, CT scan, undestructive biomechanical testing, undecalcified histology, and histomorphology. Results The fusion rates of SF/CPC, CPC/rhBMP-2, SF/CPC/rhBMP-2 and autogenous bone assessed by manual palpation were 0, 33.33%, 55.56% and 77.78% respectively at 6 months. At 12 months, the fusion rates improved to 11.11%, 44.44%, 77.78% and 77.78%, respectively.The biomechanical results showed that fusion stiffness was significantly greater in autograft compared with SF/CPC/rhBMP-2, CPC/rhBMP-2, and SF/CPC in 4 degrees of freedom (flexion, extension, right bending, and left bending) at 6 months. The SF/CPC/rhBMP-2 composite showed similar stiffness as autograft, which was significantly greater than CPC/rhBMP-2 and SF/CPC at 12 nonths. Both CPC/rhBMP-2 and SF/CPC/rhBMP-2 showed significantly greater stiffness at 12 months compared with that of at 6 months. The results showed that bone volume was significantly greater in autograft compared with SF/CPC/rhBMP-2, CPC/rhBMP-2, and SF/CPC at 6 months. There was significant difference among ceramic residue among SF/CPC, CPC/rhBMP-2 and SF/CPC/rhBMP-2, with SF/CPC the greatest and SF/CPC/thBMP-2 the least. At 12 months, the bone volume of SF/CPC/rhBMP-2 composite was comparable with autograft, and greater than that of CPC/rhBMP-2 and SF/CPC. The bone volume of SF/CPC, CPC/rhBMP-2 and SF/CPC/rhBMP-2 was significantly greater at 12 months than that of at 6 months. The ceramic residue of SF/CPC, CPC/rhBMP-2 and SF/CPC/rhBMP-2 were significantly decreased. Conclusion The SF/CPC/rhBMP-2 composite had excellent osteoconduction and osteoinduction, and balanced degradation and osteogenesis, which may be a kind of ideal bone grafts in spinal fusion.

BACKGROUND: High molecular materials have been proved to enhance the mechanical properties of calcium phosphate bone cement, as well as attenuate the injectability of composite materials. It thereby influences the clinical application of composite materials.OBJECTIVE: To observe the compressive strength and injectability of silk fihroin compound calcium phosphate bone cement, to evaluate the effect of silk fibroin on calcium phosphate, and to investigate the feasibility of applying silk fibroin as an injectable hone substitute to repair hone defects.DESIGN, TIME AND SETTING: Controlled study was performed in the central laboratory of Analysis and Testing Center, Soochow University from September to December in 2007.MATERIALS: Calcium phosphate cement was purchased from Shanghai Rebone Biomaterials Co., Ltd; silk fibroin was offered by Institute of Material & Engineering, Soochow University.METHODS: Six groups were set with different mass fractions (0.5%, 1%, 1.5%, 2%, 2.5%, 3%) of silk fibroin, which were mixed with bone cement at a certain liquid/solid ratio of 0.4 mL/g to prepare the calcium phosphate composite. The calcium phosphate cement without silk fibroin was served as control group.MAIN OUTCOME MEASURES: The compressive strength and injectability were determined. The characteristic microstructure was observed using scanning electron microscope.RESULTS: The compressive strength increased firstly and then decreased with the addition of silk fibroin. The compressive strength of the experimental groups was remarkably higher than the control group when the silk fibroin content was 1%-2.5% (P<0.05). The injectability of the paste diminished with the addition of silk fibroin, which was statistically different when the silk fibroin content was 1.5%-3% (P<0.05). Scanning electron microscope result revealed that the silk fihroin penetrated throughout calcium phosphate crystals, which were tightly connected.CONCLUSION: Silk fibroin can improve the compressive strength of silk fibroin/calcium phosphate cement composites without significant influence of manipulation, and can widen the application field of calcium phosphate bone substitute.