To set up a new rapid method for the rapid determination of influenza virus H5N1 and H7N9 basing on the Multi-Analyte Suspension Array (MASA) technology. Sequence analysis and design of degenerate primers and specific probes were set in the comparison and analysis of H5, N1, H7 and N9 genes. In combination with MASA technology, these primers and probes were used for the determination of samples of H5N1 and H7N9 and other subtypes ( H1N1, PH1N1, H5N2, H3N2 and H9N2). We developed a rapid determination method. This method had high specificity and sensitivity that could detect H5N1 and H7N9 at one time, and could detect samples that containing 10 copies of H5N1 and H7N9. This determination method could be used for rapid determination of influenza virus H5N1 and H7N9 at one time.
Humans ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza A Virus, H7N9 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; virology ; Oligonucleotide Array Sequence Analysis ; methods

Objective To construct 1.3-fold-overlength infectious clone of hepatitis B virus isolated from Chinese patients , observe the expression of plasmid in Huh7 of liver cancer cells and establish genome of HBV in vitro. Methods HBV DNA in serum was extracted from HBV patient. SOE-PCR was performed to produce a 1.3-fold-overlength genome of HBV. The plasmid was named pHBV1.3 (C). After that,pHBV1.3 (C) was transfected into Huh7 cells, HBV related viral antigens and DNA were detected by ELISA,Western Blot and Fluorescence quantitative PCR. Furthermore, adefovir dipivoxil, a clinic anti-viral drug, was utilized to test the sensitivity of the new infectious clone. Results An infectious clone of pHBV1.3 (C) was successfully constructed. HBV gene carried in pHBV1.3 (C) could be efficiently replicated and expressed in Huh7 cells. Adefovir could inhibit HBV replication in this HBV cell model. Conclusions A recombinant plasmid containing 1.3-fold-overlength of HBV genotype C was successfully constructed. This construct is competent to support viral transcription and replication in vitro , suggesting that infectious cells are expected to be a new model of HBV infection in vitro.

Objective To investigate insula cortex activation evoked by stimulating different acupoints of lower extremity.Methods Twenty healthy male right-handed subjects were recruited to receive functional magnetic resonance imaging(fMRI)examination with a whole-body 3T Siemens Trio system for whole-brain imaging.Participants were at rest during scanning and were instructed to relax with their eyes closed,without falling asleep.A blockdesigned method was applied for acupoint stimulation.The examined acupoints on the right lower extremity were stimulated,respectively,resulting in 4 experimental conditions.Stimulation was performed by brushing the subject's skin over each acupoint with a sponge at a frequency of approximately 2 Hz.Images were analyzed using SPM5 software package.Group analysis was performed with the mixed-effects model.Small volume correction was used to investigate insula cortex activation under each condition at a threshold of P ＜ O.01,with a 6-mm radius centered on the local maxima of activation in insula cortex.The scope of insula cortex was determined with SPM anatomy toolbox.Results All the examined acupoints induced activation in contralateral insula cortex.Activation clusters evoked by different acupoints presented various types of locations and shapes,and all of them located in the posterior part of insula cortex.Activation in the ipsilateral insula cortex was not found for all examined acupoints.Conclusions Stimulating lower extremity acupoints can activate posterior insula cortex.This effect may promote the recovery of dysphagia via several mechanisms including facilitating the initiation of swallowing,improving oral and pharyngeal sensation,and reducing taste disturbance.

Background One of the features of the pathogenesis of primary angle-closure glaucoma(PACG) is anterior synechia of peripheral iris.Goniosynechialysis and combination of phacoemulsification and goniosynechialysis have been applied for the treatment of the disease recently,but the selection of operative types has great impact on clinical efficacy.Objective This study was to investigate the long-term efficacy of goniosynechialysis and combination of phacoemulsification and goniosynechialysis for the management of chronic PACG.Methods A non-randomized clinical controlled trial was designed.This clinical trial complied with Declaration of Helsinki and was approved by Medical Ethic Committee of Nanjing Medical University.Written informed consent was obtained from each patient.One hundred and ten eyes of 110 patients with chronic PACG were assigned to the goniosynechialysis group and combined operative group from March,2008 to February,2011 in Wuxi People's Hospital.180° goniosynechialysis was performed on 34 patients in the goniosynechialysis group,and phacoemulsifieation +intraocular lens (IOL) implantation + goniosynechialysis were carried out in 78 patients of the combination operative group.All the patients were followed-up for 2 years.Vision acuity,intraocular pressure (IOP),anterior chamber depth(ACD),unltrasound biomicroscopy and perimetry were recorded and compared between before and after operation.Results No significant difference was found in vision acuity between preoperation and postoperative 2 years in the goniosynechialysis group ([0.65 ± 0.15] vs.[0.45 ± 0.15]) (t =1.57,P＞0.05),but in the combination operative group,the vision acuity was significantly different between the before and after operation ([0.25±0.15] vs.[0.85 ±0.05]) (t =9.12,P＜0.001).The lOPs at 2 years after operation were (14.2±4.1) mmHgand(13.7±4.8) mmHg,respectively in the goniosynechialysis group and combination operative group and were significantly lower than(47.2 ±6.3) mmHg and(46.9±7.0) mmHg before operation(t =4.95,P＜0.001 ;t=5.03,P＜0.001).The ACD values in the goniosynechialysis group and combination operative group were(3.38±0.02)mm and (3.54±0.03) mm 2 years after operation,which were significantly increased in comparison with (1.33 ±0.24)mm and (1.56±0.37) mm before operation(t=7.65,P＜0.001;t=6.76,P＜0.001).Conclusions Both combination of phacomulsification with goniosynechialysis or 180° goniosynechialysis are effective for the treatment of PACG.Suitable operation should be alternated depending on the indicators of PACG patients.

Objective To explore the effect and mechanism of electroacupuncture at Baihui(GV20) and Zusanli(ST36) of patients with depression.Methods90 patients were randomly divided into group A electroacupunctured at Baihui(GV20) and Zusanli(ST36), group B electroacupunctured at Taichong(LR3), Sanyinjiao(SP6), Neiguan(PC6), and Shenmen(H7) and group C treated with Fluoxetine (20 mg/d, 6 weeks). The therapeutic effect was assessed by the Hamilton Depression Rating Scale (HAMD) before and after treatment. Serum cytokine levels (IL-2, IL-4, and IFN-γ) were measured by ELISA assay at admission and after treatment (6 weeks).ResultsCompared to pre-treatment, the mean levels of HAMD ( P<0.01)and IL-4 (P <0.01) decreased significantly in all groups after treatment. Rank distribution of HAMD scores had obvious difference between all groups. The mean levels of IL-2 and IFN-γ had no significant difference in group C after treatment (P>0.05). And the mean levels of IL-2 and IFN-γ increased significantly in groups A and B after treatment(P<0.01, P<0.05).ConclusionElectroacupuncture takes effect earlier than fluoxetine for depression. Electroacupuncture at Baihui(GV20) and Zusanli(ST36) was more effective than the other two methods. The three types of treatments can restore three types of cytokines and improve depressive syndrome.

Objective To investigate the levels of serum vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in patients with non-small cell lung cancer(NSCLC) and relationships with c1inicopatho1ogica1 characteristics and their clinical significance. Methods The concentrations of serum VEGF and bFGF were detected by enzyme-linked immunosorbent assay(ELISA) in 40 patients with NSCLC before and after chemotherapy. Results The level of serum VEGF in patients with Ⅳ stage NSCLC was significantly higher than that of Ⅲ stage(P

Objective To analyze the association of the morbidity,the management of blood glucose,and the prognosis of patients with hyperglycemia in the medical intensive care units(ICU).Methods Medical records of ICU patients of Renji Hospital from 2002 to 2009 were reviewed using Medical Record Inquiry System,and the data were retrospectively analyzed.Results(1)2631 subjects were included in the present study,blood glucose was determined at least once during hospitalization in 2168 of them.The incidence of hyperglycemia was 26.3%,in which 12.9% presented a known history of diabetes and 13.4% without.In the patients with diabetes history,93.2% of them received anti-diabetic treatment during hospitalization.mainly with oral anti-hyperglyeemic agents (53.0%)or subcutaneous insulin injection(24.9%).However,in the patients without diabetes history,84.4% were not treated against hyperglycemia.The mortality was increased in the latter group(30.4% vs13.9%,P<0.01).(2)In the patients with diabetes history,the mortality in patients whose blood glucose>10 mmol/L was higher than those with blood glucose≤7.0 mmol/L(20.5% vs 9.9%,P<0.05):while in the patients without diabetes history,the mortality began to rise as blood glucose>7.0 mmol/L(P<0.01).(3)Multiple stepwise regression analysis revealed that the average blood glucose level was an independent risk factor for death(OR=1.26).Conclusions The ICU patients showed a high prevalence of hyperglycemia,the management of hyperglycemia should be emphasized.Hyperglycemia in critically ill patients might be an independent risk factor of increased mortality.

Objective To validate the clinical effect of evidence-based nursing on early rehablilitation of patients with acute myocardial infarction undergoing PTCA and intracoronary stenting. Methods 47 patients with acute myocardial infarction undergoing PTCA and intracoronary stenting were divided into the nursing group(24 cases)and the control group(23 cases).The nursing group received nursing with evidence-based nursing pattern while the control adopted routine nursing method. The nursing effect in the two groups was compared in the following aspects such as the incidence rate of adverse effect,average hospitalization days, athletic ability and psychological status. Results The incidenece rate of adverse effect Was lower(P＜0.05)and the average hospitalization days were shorter(P＜0.01)than those of the control group with addition of cardiovascular complications(P＞0.05).The athletic ability and psychological status were also superior to those of the control group(P＜0.05).Conclusion Application of evidence-based nursing could hasten the rehabilitative process of patients undergoing PTCA and intracoronary stenting.

By skin prick test,three kinds of mite allergens(Dermatophagoides pteronyssinus,Dermatophagoides farinae,Blomia tropicalis)were tested in a group of asthma children in Jinhua area from Oct 2005 to Sep 2006.The pos-itive rate to allergen from D.pteronyssinus and D.farinae was 80.6% and 77.8% respectively,higher than that of Blomia tropicalis(61.1%)(?2=21.39,P

AIM: To study the isolation,expansion and purification of mesenchymal stem cells(MSCs) from human umbilical cord blood(UCB),and investigate some biological identities of MSCs.METHODS:(1) MSCs of UCB,adult bone marrow(BM) and fetus BM were isolated by centrifugation with Ficoll,and the different kinds of MSCs were observed everyday.(2) Surface markers of MSCs were identified by flow cytometry.(3) The level of HGFs(TPO,SCF,FLT-3L,IL-6) secreted by different sources of MSCs was checked by ELISA method.RESULTS:(1) No difference in morphology of the colonies between UCB MSCs and BM MSCs was observed.However,the mononuclear cells needed in culture of UCB MSCs was about 3 times more than that in culture of BM MSCs.The times of UCB MSCs colony formation and confluencing were longer than that of BM in primary culture.(2) After passaged,there was no significant difference in the proliferation rates of 3 kinds of MSCs.Only 4 of 15 UCB samples contained a homogeneous population of MSCs.(3) UCB MSCs shared the same markers with BM MSCs.Neither hematopoietic marker nor immunologic recognition antigens were expressed.(4) The level of hematopoietic growth factors(HGFs) secreted by 3 kinds of MSCs was similar.CONCLUSIONS:(1) MSCs were isolated from UCB,but the amount of MSCs in UCB was smaller than that in BM,and just seldom samples of UCB contained homogeneous MSCs.(2) MSCs from UCB and BM shared the same biological characteristics,such as proliferation ability,surface markers,immunophenotypes and HGFs secretion.