Objective To construct recombinant full length genotype B and C hepatitis B virus (HBV)and to examine HBV DNA replication and hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)expressions in Huh7 cells. Methods The full length genotype B and C HBV DNA were extracted and amplified from two HBV infected patients. The recombinant plasmids were constructed by inserting the amplified HBV fragments into the eukaryotic expression vector,pHY106,which were then transfected into Huh7 cells. The cells transfected with blank pHY106 vector were used as control. HBV DNA replication at 72 hours of transfection was detected by Southern blot. The HBV DNA levels in Huh7 cells at 24,48,72,96 and 120 hours of transfection were determined by real-time polymerase chain reaction(PCR).Meanwhile, the HBsAg and HBeAg expression levels in the supernatants at 24,48,72,96 and 120 hours were determined by enzyme linked immunosorbent assay(ELISA).Results The recombinant plasmids expressing genotype B or C HBV DNA were successfully constructed.The HBV replicative intermediates in HBV core particles,including rcDNA dsDNA and ssDNA,were detected by Southern blot.HBV DNA level could reach 8 lg copy/mL which was by real-time PCR. HBsAg and HBeAg levels determined by ELISA peaked at 72 hours after transfection and then declined gradually. Conclusions The recombinant plasmids inserted with genotype B or C HBV DNA are constructed successfully, which can express high levels of HBsAg and HBeAg in Huh7 cells. This system provides a platform for studying the pathogenesis of B and C genotype HBV, the interaction between HBV and host, as well as exploiting new drugs against HBV.

Adult ; Bronchoalveolar Lavage ; methods ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; therapy ; Quinuclidines ; therapeutic use

Objectives To validate the Screening Tool for the Assessment of Malnutrition in Pediatrics (STAMP) in hospitalized pediatric patients, and to assess the nutritional status and nutritional risk using STAMP. Methods the nutritional status of hospitalized pediatric patients was investigated by adopting STAMP. The STAMP score≥4 was deifned as a high standard of nutritional risk. Results Among 1 506 hospitalized children, children with high nutritional risk accounted for 26.56%. The overall prevalence of malnutrition and stunting was 19.12%and 10.16%, respectively. There was signiifcant diffe-rence in stunting among each department (P<0.05), which was the highest in department of internal pediatrics. The proportion of patients receiving nutritional support was 10.09%. Among them, the rate of parenteral and enteral nutrition support was 7.84%and 2.92%, respectively. The ratio of parenteral nutrition to enteral nutrition was 2.68:1. After intervention to hospitalized pa-tients with high nutritional risk, the overall prevalence of high nutritional risk decreased from 26.56%at admission to 21.71%at discharge (P<0.05). Conclusions STAMP can objectively relfect the possible malnutritional risk in hospitalized pediatric patients. The nutrition condition of the pediatric patients can be improved through nutritional risk management.

There are disadvantages in examination in basic medical stage. For example, there is too much quantity of work in written examination and the time is short for preparation for oral examination. Basic medical college reforms the examination in the first stage. The exam is composed of two parts. One is performed in computer instead of being written on paper,the oth-er is oral test whose topic is informed one term ahead of time. These can improve the students’ ability to summarize what they have learned in class.

Objective: To investigate the cognitive impairment features in patients with amnestic mild cognitive impairment(aMCI) and vascular cognitive impairment-no dementia(VCI-ND).Methods:Sixteen normal elders,10 patients with aMCI and 12 patients with VCI-ND were recruited.The normal elders were selected from communities in Shanghai,while the aMCI and VCI-ND patients were selected from outpatient clinic.All participants ranged in age of 50～80 years,with education level of junior middle school or above,and they completed cranial CT or MRI and a series of neuropsychological tests.Results:In the three memory tests,aMCI group performed worst.The scores of both aMCI and VCI-ND groups were lower than that of the normal elders.For example,the delayed recall scores of the Rey-Osterrich complex figure test in the three groups were(18.8?9.5)(normal),(5.6?5.6)(aMCI) and(9.6?7.0)(VCI-ND)(P

Objectives To screen the differentially expressed proteins in serum of population chronically exposed to arsenic in drinking water,thus to provide candidate protein biomarkers for arsenic exposure and arsenicosis.Methods Subjects were selected from the drinking water type of endemic arsenicosis areas in Shanxi province,China.Demographic characteristics,history of arsenic exposure,cigarette smoking,alcohol drinking,health and other information were collected using questionnaire.The subjects were divided into low-arsenic group (with arsenic in drinking water ＜ 10 μg/L),medium-arsenic group( 10 - 50 μg/L),high-arsenic group( ＞ 50 μg/L),and arsenicosis group(the drinking water with arsenic ＞ 50 μg/L was replaced by low arsenic water ＜ 10 μg/L).The number of cases in each group was 30.The arsenicosis patients were diagnosed according to “Standard of Diagnosis for Endemic Arsenism” (WS/T 211-2001 ).With the principle of informed consent,blood samples were collected.Differentially expressed serum proteins of different arsenic exposure groups and arsenicosis group were screened by two-dimensional differential gel electrophoresis(2-D DIGE),and further identified by mass spectrometry (MS).Results An average of (1299 ± 167) protein spots were identified in 6 gel images and 688 protein spots were discovered repeatedly in at least 5 gels.There were 33 protein spots differentially expressed among low-,medium- and high-arsenic groups P ＜ 0.01).Fifty four protein spots were significantly different among low-,medium-,high-arsenic exposure groups and arsenicosis group(P ＜ 0.01 ).Twenty five protein spots were selected for MS analysis,and13 protein spots were identified.Compared with low-arsenic group,the expressions of apolipoprotein A-Ⅳ,retinol binding protein,and estrogen receptor hypothalamic isoform in medium- and higharsenic exposure groups were down regulated,and the expressions of component 4A and 4B were up regulated.Compared with low-,medium- and high-arsenic groups,the expressions of beta-2-glycoprotein Ⅰ,Keratin 1,hemopexin,complement C1r subcomponent,and ficolin-3 in arsenicosis group were down regulated,and the expressions of pigment epithelial-differentiating factor,alpha-1-microglobulin and carboxypeptidase N catalytic chain were up regulated.Conclusions Chronic arsenic exposure can significantly change population's serum protein expression.Differentially expressed proteins in arsenicosis patients will not decline with the decline of arsenic in a short term.Whether or not the differentially expressed proteins identified in this study can be used as biomarkers for arsenic exposure and arsenicosis needs to be further verified.

Objective To construct prokaryotic expression vector carrying jc virus(JCV)t-antigen gene,express and purify this fusion protein.Methods The JCV t-antigen gene from a cerebrospinal fluid sample was amplified using polymerase chain reaction(PCR)method.After sequencing.the gene was cloned into plasmid pET32a(+)to construct recombinant prokaryotic expression vector pET32a(+)-t.The t-antigen fusion protein was expressed by isopropy-~D-thiogalactoside(IPTG)induction and prepared in large scale,then purified by Ni+affinity column chromatography.The polyclonal antibody was obtained from the BAI.B/C mouse immunity by the purified protein.Results The relative molecular nlass of recombinant protein expressed by pET32a(+)-t was about 41 000.Sodium dodeeylsulfate-polyaerylamide gel electrophoresis(SDS-PAGE)showed that the fusion protein W&S highly expressed after 3.5～20.Oh of IPTG induction.The antigenicity of the purified protein Was well confirmed by Western blot.The anti-mousepolyclonal antibody was obtained successfully from immunized BALB/c mice.Conclusions The prokaryotic expression vector pET32a(+)-t is successful constructed and the fusion protein is expressed and purified.Furthermore,the antibody of JCV small envelop protein t is successfully prepared.This work provides vMuable information for further study on epidemiology and biological function of t antigen.

Acclimatization ; physiology ; Adult ; Altitude ; Fatty Acid-Binding Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Myelin Basic Protein ; metabolism ; Nervous System Diseases ; metabolism ; Occupational Exposure ; Succinate Dehydrogenase ; metabolism

Objective To evaluate the effect of BML-111 on NF-κB pathway during acute lung injury induced by hemorrhagic shock and resuscitation in rats.Methods Thirty-two adult male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 4 groups (n =8 each) using a random number table:sham operation group (group S),hemorrhagic shock and resuscitation group (group HSR),BML-111 group,and BML-111 + BOC-2 (lipoxin A4 receptor antagonist) group (group BOC-2).The animals were anesthetized with intraperitoneal pentobarbital sodium.Hemorrhagic shock was induced by blood letting and maintained for 30 min.The animals were then resuscitated for 30 min by infusion of the shed blood and lactated Ringer's solution.In group BOC-2,BOC-2 (50 μg/kg) was injected intraperitoneally before blood letting.In BML-111 and BOC-2 groups,BML-111 (1 mg/kg) was injected intraperitoneally at the beginning of resuscitation.The rats were sacrificed at 2 h after the end of resuscitation and lungs were removed for determination of pathological changes,myeloperoxidase (MPO) activity,intercellular adhesion molecule-1 (ICAM-1) expression (by immunohistochemistry),tumor necrosis factor-alpha (TNF-α) content (by ELISA),and NF-κB p65 and IκB-α expression (by Western blot).Results Compared with group S,the MPO activity,ICAM-1 expression,and TNF-α content were significantly increased,NF-κB p65 expression was up-regulated,and IκB-α expression was down-regulated in group HSR.Compared with group.HSR,the MPO activity,ICAM-1 expression,and TNF-α content were significantly decreased,NF-κB p65 expression was down-regulated,IκB-α expression was up-regulated,and pathological changes of lung were attenuated in group BML-111.Compared with group BML-111,the MPO activity,ICAM-1 expression,and TNF-α content were significantly increased,NF-κB p65 expression was up-regulated,and lκ:B-α expression was down-regulated,and pathological changes of lung were aggravated in group BOC-2.Conclusion BML-1 11 inhibits activation of NF-κB pathway and inflammatory responses,thus mitigating acute lung injury induced by hemorrhagic shock and resuscitation in rats.

OBJECTIVE To investigate the uses of antibacterials,the characteristics of bacterial distribution and the drug resistance and provide a foundation for reasonable application of antibacterials.METHODS Germs isolation and cultivation were carried out according to National Clinical Testing Operation Procedures(2nd edition).Germs definition and resistance test were carried out by using VITEK 2 Compact microbiological assay system.Statistical analysis was processed by using WHONET 5.4.RESULTS Totally 7815 strains of bacteria were isolated from clinical samples,most of which were Gram-negatives(G-)(58.0%,56.3% and 59.5%,respectively in the 3 years).The tendency of Gram-positives(G+) didnot charee obviously(about 20%).Theresistance were increasing.The resistance rate of Staphylococcus aureus was more than 70.0 % to the most antibacterials.Up to now,there was no vancomycin-resistant isolate.CONCLUSIONS We should use antibacterials reasonably and efficiently to delay and control drug resistance,according to the susceptibility test.