Objective To study the immunoreaction of the recombinant proteins encoded by the fragments of ORF2 ( second open reading frames) gene of hepatitis E virus ( HEV). Methods The aimed sequence from the full-length ORF2 in clone PEH2 , which was derived from a Chinese strain of HEV,was amplified and cloned it into vector pcDNAS. 1 which was then transfected to 293 cell. Results The ORF2 protein was present in the soluble fractions of the cell lysate. The expressed protein of HEV ORF2 in 293 cells by using a plasmid pcDNA3. 1-based system showed positive on immunoblots probed against antibodies raised in BALB/c mice. Conclusion The experimental results laid a foundation for developing diagnostic reagent to detect HEV by using the expressed products of ORF2 protein.

Filamentous fungi have been used in industrial fermentation extensively. Based on non-phosphorylating electron transport process, alternative respiration pathway (ARP) acts as an energy overflow, which can balance carbon metabolism and electron transport, allow the continuance of tricarboxylic acid cycle without the formation of ATP, and permit the turnover of carbon skeletons. Alternative respiration pathway also plays an important role in the stress response of fungi and the physiological function of conditioned pathogen. Alternative oxidase (AOX) is the terminal oxidase responsible for the activity of alternative respiration pathway, which exists widely in higher plants, parts of fungi and algae. Owing to the property that alternative oxidase (AOX) is sensitive to salicylhydroxamic acid (SHAM) and insensitive to conventional inhibitors of cytochrome respiration, alternative respiration pathway by AOX is also named as cyanide-resistant respiration (CRR). In recent years, the study of the alternative respiration pathway and alternative oxidase has been a hot topic in the area involving cellular respiration metabolism. In this review we summarized the latest research advances about the functions of alternative respiration pathway and alternative oxidase in industrial fungi.
Electron Transport ; Fermentation ; Fungi ; enzymology ; Industrial Microbiology ; Mitochondrial Proteins ; chemistry ; Oxidoreductases ; chemistry ; Plant Proteins ; chemistry

Objective To explore whether human umbilical venous endothelial cells could be used as feeder layer to support the growth of embryonic stem cells (ESC) and keep ESC undifferentiated.Methods The venous vessels of umbilical cord obtained from healthy puerperal were perfused with collagenase.The isolated endothelial cells went through primary culture and passages for expansion.Factor Ⅷ antigens determination was implemented.Endothelial cells with good growth and 3 or above passages were treated with mitomycin-C(10 mg/L) and prepared as feeder layer,on which E14.1 ESC was transplanted for subculturing to observe the morphological characterization and determine ESC alkaline psphatase (AKP) activity and the expression of stem cell marker Oct-4.Severe combined immune deficiency(SCID) mouse in vivo terotoma formation experiment was performed to identify its pluripotent properties.Results Human umbilical vein-derived endothelial cells grew well in culturing in vitro and regenerate in large numbers.The endothelial cells maintained normal cellular morphological and biological characterization after 10 passages.The cells stopped proliferating after being treated with mitomycin-C,but its activity and morphological properties were well-maintained with 24 hours,which was a fundamental property of serving as feeder layer.E14.1 ESC remained undifferentiated in human umbilical venous endothelial cells after 3-8 passages,the cells grew in colony and showed high expression of AKP and stem cell Oct-4.In vivo pluripotency experiment showed that 6 weeks after being transplanted to SCID mice E14.1 ESC of 6 and 10 passages in endothelial cells both could form teratoma containing 3 layers of tissue cells.Conclusions Human umbilical venous endothelial cell serve as a convenient feeder layer cell with rich sources.It can effectively support ESC growth and heterogenous and prevent the heterogeneous protein pollution and pathogenic microorganisms caused by animal cell feeder layers,thus solve the problem of biological safety of ESC clinical application.

Objective To investigate the serum levels of IL-10 and TGF-β1 and their significance for patients with sys-temic lupus erythematosus( SLE) .Methods A total of 135 cases were available for the study including 63 cases in primary group(A), 40 cases in remission group(B) and 32 cases in healthy control group(C).ELISA method was used to deter-mine the concentrations of IL-10 and TGF-β1 in serum.Finally, the results of group A were analyzed on the basis of System-ic Lupus Erythematosus Disease Activity Index( SLEDAI) .Results The level of IL-10 in serum of group A was significant-ly higher than that of group B and group C.The level of IL-10 in serum of group B was significantly higher than that of group C(P<0.05).The level of IL-10 in group A had active correlation with SLEDAI.The level of TGF-β1 in serum of group A was significantly lower than that of group B and group C.There was no statistical difference between group B and group C(P>0.05).The level of TGF-β1 in serum of group A had negative correlation with SLEDAI.The cases in group A were divided into three subgroups by SLEDAI:21 cases in mild activities subgroup, 30 cases in moderate activities sub-group,and 12 cases in severe activities subgroup, the levels of IL-10 and TGF-β1 in the three subgroups were significantly different(P<0.05).Conclusions IL-10 and TGF-β1 are involved in activity and severity of SLE.

Objective To investigate the influences of survivin down-regulation on cell G2/M phase arrest,apeptosis and sensitivity to carbon ion irradiation. Methods Small interfering RNA (siRNA) targeting survivin mRNA was designed, in vitro chemo-synthesized and transfected into SMMC-7721 cells. Survivin mRNA expression in SMMC-7721 cells was measured by real-time PCR, and the apeptotic rates by Annexin-FTTC at 24 and 48 h after transfection. Cell G2/M phase arrest after transfection was assessed with flow eytometry as well. Cellular sensitivity to high-LET carbon ions was determined by means of colony-forming assay. Results The expressions of survivin at mRNA level were down-regulated to be 59% and 39% in relation to the non-treated cells at 24 and 48 h after siRNA transfeetion, respectively. G2/M phase arrest in SMMC-7721 cells at 24 h after transfection was observed while much more obvious at 48 h. The apeptotic rate of SMMC-7721 cells was 21.41 % at48 h after survivin siRNA transfection, which was significantly higher than that of the cells transfected with negative siRNA. Moreover, a decreased clonogenic survival in siRNA treated group was shown. Conclusion Down-regulation of survivin gene expression in SMMC-7721 cells by siRNA could effectively induce cell apeptosis and G2/M phase arrest, and enhance the cellular radiosensitivity to high-LET heavy ions.

Objective To prepare a human meningococcal reference serum and standardize IgG concentrations to capsular polysaccharides and in vitro bactericidal activities of the reference serum against serogroup A, C, Y and W135 strains.Methods Twenty healthy adults were recruited and given one dose of immunization with tetravalent (serogroups A, C, Y and W135) meningococcal polysaccharide vaccine . Plasma samples were collected and gone through a series of process treatments including defibrination , filtra-tion, and lyophilization to prepare the meningococcal reference serum Men 10.The IgG concentrations of Men10 to capsular polysaccharides of serogroups A , C, Y and W135 were calibrated by using an internation-al reference serum CDC1992 as the standard in enzyme-linked immunosorbent assay (ELISA).Provisional IgG concentrations of Men10 were intensively validated by testing a panel of 12 calibration serum samples from Centers for Disease Control and Prevention , USA ( US CDC) and a panel of 56 serum samples immu-nized with A, C, Y and W135 meningococcal polysaccharide vaccine from Lanzhou Institute of Biological Products Co., Ltd.(LIBP) with the assays using Men10 and CDC1992 as the standard and/or test sam-ples, respectively.The bactericidal titers against serogroup A , C, Y and W135 strains were measured by se-rum bactericidal assay (SBA).Results Four thousand vials (0.5 ml/vial)of lyophilized human meningo-coccal reference serum Men10 were successfully prepared with 2.5%of residual moisture .Reference serum Men10 was sterile and free from contamination by hepatitis B virus , hepatitis C virus , human immunodefi-ciency virus and syphilis .Provisional IgG concentration of Men 10 to capsular polysaccharide of serogroups A, C, Y and W135 was calibrated by using CDC1992 as the standard.Furthermore, IgG concentrations of both panels of 12 CDC calibration serum samples and 56 LIBP serum samples calibrated by using Men 10 as the standard correlated well with those by using CDC1992 as the standard (r=0.99,P<0.05).The IgG concentrations of CDC1992 as calibrated by using Men10 as the standard showed significant correlation with its previously determined values with variation <10%.SBA titers for serotype A , C, Y and W135 strains were established as well .Conclusion A panel of new human meningococcal reference serum Men 10 with accurately calibrated IgG concentration against capsular polysaccharide of serogroups A , C, Y and W135 as well as SBA titers was successfully established .

This study aims to analyze the molecular epidemiological characteristics of HIV-1 strains prevailing among men who have sex with men (MSM) in Beijing, China. The pol gene fragments from 250 newly diagnosed HIV-1-infected MSM individuals during 2006-2010 in Beijing were amplified by RT-nested PCR, sequenced, and phylogenetically analyzed. HIV-1 pol gene from 189 individuals were amplified and analyzed; 81 (42. 9%), 3 (1. 6%), 2 (1.0%), 88 (46. 6%), and 15 (7.9%) individuals were infected with HIV-1 subtypes B, B', C, CRF01_AE, and CRF07_BC, respectively. The subtypes B and CRF01_AE could both be grouped into two clusters, and CRFO7_BC strains shared high homology and were presumed to originate from a common ancestor. The HIV-1 circulating in MSM in Beijing had a lower genetic diversity than in heterosexuals. The HIV-1 epidemic (2006-2010) in MSM in Beijing was actually a rapid spread of HIV-1 CRF01 AE and B, or rather native strains of the two viruses.
Adult ; China ; epidemiology ; Epidemics ; Genetic Variation ; HIV Infections ; diagnosis ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Young Adult

beta-CIT was labeled with 131I by the peracetic acid method. Cat model of Parkinsonism was set up with MPTP. Each of normal and PD model cats was given an injection of 74 MBq/0.5 ml 131I-beta-CIT into the femur vein. Then the blood samples were obtained at 4 h and 20 h, the radioactivity was counted with calibrator. The biodistribution data of 131I-beta-CIT in cat body was calculated (ID%/g). The cats were subjected to imaging at 0.5 h, 1 h, 2 h, 4 h, 20 h after the administration of radiopharmaceutical. The radioactivity in striatum and cerebellum was measured and striatal specific binding ratios were calculated. The Results showed that the radio chemical purity of 131I-beta-CIT was 97.62% +/- 0.31%. The 131I-beta-CIT remained stable for at least 4 h after incubation with water and serum respectively. Following intravenous administration in cats, 131I-beta-CIT showed high accumulation in striatum. The study of imaging in cats showed that striatal specific uptake of 131I-beta-CIT at 20 h after injection was 4.83 +/- 0.82 in normal cats and 2.92 +/- 0.66 in PD cats. There was a significant reduction of striatal tracer uptake in PD cats, compared to the controls. The results of biodistribution study was in agreement with the results of imaging study. These results suggest that beta-CIT is an ideal agent for dopamine transporter imaging and can be used for the diagnosis of Parkinson's disease.
Animals ; Brain ; metabolism ; Cats ; Cocaine ; analogs & derivatives ; metabolism ; Disease Models, Animal ; Dopamine Plasma Membrane Transport Proteins ; Female ; Male ; Membrane Glycoproteins ; metabolism ; Membrane Transport Proteins ; metabolism ; Mice ; Mice, Inbred Strains ; Nerve Tissue Proteins ; metabolism ; Parkinson Disease ; diagnostic imaging ; metabolism ; Tomography, Emission-Computed, Single-Photon

Objective To investigate the efficacy of lateral decubitus intramedullary nailing for treatment of subtrochanteric fractures of the femur.Methods From January 2012 to December 2015,23 patients with simple femoral subtrochanteric fracture were treated at Department of Orthopedic Trauma,Changhai Hospital.They were 15 males and 8 females,aged from 19 to 77 years (average,48.3 years).According to the Seinsheimer classification,there were 6 cases of type ⅡB,8 cases of type ⅡC,6 cases of type Ⅲ A,and 3 cases of type ⅢB.Their injuries were caused by traffic accident in 10 cases,falling from a height in 5 cases,and sprain in 8 cases.All patients were treated by closed reduction and anterograde intrarnedullary nailing at lateral decubitus.Their operative time,bleeding volume,fluoroscopic frequency,fracture healing time,functional recovery and complications were recorded and analyzed.Results Their operative time ranged from 55 to 80 min,averaging 65.7 min;their bleeding volumes ranged from 240 to 420 mL,averaging 304.3 mL;their fluoroscopic frequency ranged from 30 to 60 times,averaging 42.7 times.This cohort was followed up for 12 to 28 months (average,17.9 months).Their fracture healing time ranged from 4 to 10 months,averaging 5.5 months.Nonunion occurred in one patient but was cured by secondary operation.The HSS evaluation at the final follow-ups showed 17 excellent cases and 6 good ones,yielding an excellent to good rate of 100％.All the wounds healed by the first intention.No infection,deep vein thrombosis or implant failure was observed.Conclusion As lateral decubitus intramedullary nailing can achieve satisfactory clinical efficacy for subtrochanteric fractures of the femur,the body position of lateral decubitus may be a good alternative.

Donation after brain death followed by circulatory death (DBCD) is a unique practice in China. The aim of this study was to define the pathologic characteristics of DBCD liver allografts in a porcine model. Fifteen male pigs (25-30 kg) were allocated randomly into donation after brain death (DBD), donation after circulatory death (DCD) and DBCD groups. Brain death was induced by augmenting intracranial pressure. Circulatory death was induced by withdrawal of life support in DBCD group and by venous injection of 40 mL 10% potassium chloride in DCD group. The donor livers were perfused in situ and kept in cold storage for 4 h. Liver tissue and common bile duct samples were collected for hematoxylin and eosin staining, TUNEL testing and electron microscopic examination. Spot necrosis was found in hepatic parenchyma of DBD and DBCD groups, while a large area of necrosis was shown in DCD group. The apoptosis rate of hepatocytes in DBD [(0.56±0.30)%] and DBCD [(0.50 ± 0.11)%] groups was much lower than that in DCD group [(3.78±0.33)%] (P<0.05). And there was no significant difference between DBD group and DBCD group (P>0.05)). The structures of bile duct were intact in both DBD and DBCD groups, while the biliary epithelium was totally damaged in DCD group. Under electron microscope, the DBD hepatocytes were characterized by intact cell membrane, well-organized endoplasmic reticulum, mild mitochondria edema and abundant glycogens. Broken cell membrane, mild inflammatory cell infiltration and sinusoidal epithelium edema, as well as reduced glycogen volume, were found in the DBCD hepatocytes. The DCD hepatocytes had more profound cell organelle injury and much less glycogen storage. In conclusion, the preservation injury of DBCD liver allografts is much less severe than that of un-controlled DCD, but more severe than that of DBD liver allografts under electron microscope, which might reflect post-transplant liver function to some extent.