In the last decade, next-generation sequencing (NGS) technology, which is characterized by being high-throughput, rapid, sensitive, and accurate, has developed rapidly. Main components of NGS are platforms: 454 sequencing; illumina sequencing; ion torrent sequencing; SOLID sequencing. NGS is used widely for the human immunodeficiency virus (HIV)-1. In this review, we focus on applications of the dynamics of HIV-1 quasispecies.
Animals ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; High-Throughput Nucleotide Sequencing ; methods ; Humans

Objective T_O analyze the electrophysiological features of shoulder-hand syndrome(SHS)fol- lowing stroke and investigate the relationships between peripheral nerve damage and the factors causing SHS.Meth- ods Fifty-eight stroke patients were divided into an SHS group(39 patients with shoulder-hand syndrome)and a model group(19 patients without shoulder-hand syndrome).Standard sensory and motor nerve conduction studies were performed with 58 of the patients,including sensory nerve conduction velocity(SCV),amplitude of the sensory nerve action potential(SNAP),distal motor latency(DMI)and the amplitude of the compound muscle action poten- tial(CMAP)of the median nerve.A needle electromyogram(EMG)test was performed on the abductor pollicis of the affected side with both groups.Results The needle EMG showed abnormal insertion potential,fibrillation po- tential and positive sharp waves in all 39 cases of the SHS group,which was significantly higher than in the model group.The amplitude of the sensory nerve action potential(7.77?4.34 mV)and the amplitude of the compound muscle action potential(10.13?3.15 mV)were significantly lower than those of the model group.Abnormal ampli- tude was more severe in sensory nerves than in motor nerves.Conclusion Peripheral nerve damage was found in the shoulder-hand syndrome patients,and this damage was mainly the dystrophy of axonal neuropathy.The damage severity was more in sensory nerves than in motor nerves.The study could offer an useful clue of prevention and treat- ment of shoulder-hand syndrome.

Objectives To understand current status of the admission and treatment for the patients with community-acquired pneumonia (CAP) in central hospitals of Shanghai area,and to evaluate the severity of patients admitted to the hospital with CAP by the criteria set in the Guidelines for Diagnosis and Treatment for CAP developed by the Chinese Medical Association in 2006 and provide evidence for its popularization and application throughout the country.Methods Medical records of 137 patients with CAP admitted to the hospital from January 1,2005 to September 30,2006 were retrospectively studied and analyzed with SPSS 10.0 software.Chi-square test and ANOVA were used to evaluate the severity of the patients with CAP by the criteria set in the Guidelines and to correlate it with pneumonia severity index (PSI).Statistical analysis was performed for the difference between length of hospitalization,cost,length of intravenous use of antibiotics,the number of risk factors,and fatality during hospitalization between three groups of patients categorized based on the severity criteria in the Guidelines.Results There existed a good relationship between the criteria for severity of CAP by the Guidelines and PSI,with a Pearson's coefficient of correlation of 0.577,P

BACKGROUND: Sepsis is a tough problem in critical ill patients. This study aimed to investigate the dynamic changes of monocyte Toll-like receptor (TLR) 4 expression in peripheral blood of septic rats and to determine the effects of transforming growth factor (TGF) -β1 on TLR4 expression. METHODS: Altogether 132 clean level SD rats were randomly divided into a control group (n=12), a sepsis model group (n=60), and a TGF-β1 intervention group (n=60). In the sepsis model group and TGF-β1 intervention group, the rats were subdivided into five groups (2-hour group, 6-hour group, 12-hour group, 24-hour group, and 48-hour group), with 12 rats in each group. Cecal ligation puncture (CLP) was performed in the sepsis model group and TGF-β1 intervention group to establish models of sepsis. The rats in the sepsis model group were injected with 1 mL normal saline at the caudal vein 0.5 hour after the model establishment; the rats in the TGF-β1 intervention group were injected with 20 ng/mL or 250 g TGF-β10.5 hour after the model establishment. Flow cytometry was used to detect the change of monocyte TLR4 in peripheral blood, and enzyme-linked immunosorbent assay (ELISA) was used to detect the change of TNF-α level in peripheral blood. RESULTS: At 6-12 hours after CLP, the monocyte TLR4 in peripheral blood started to decrease, and reached the lowest level at 12 hours. Compared to the control group, the monocyte TLR4 expression at 6 and 12 hours was lowered significantly (P<0.05). Compared to the sepsis model group at 2, 24 and 48 hours after CLP, the monocyte TLR4 expression in the TGF-β1 intervention group decreased dramatically (P<0.05), but there were no differences between the two groups at 6 and 12 hours respectively. Compared to the control group, the concentration of NF-κB in liver tissue increased significantly 6 hours after CLP (P<0.05). After use of TGF-β1, the concentration of NF-κB was decreased significantly but still higher than that of the control group. Compared to the control group, the concentration of TNF-α in peripheral blood was increased significantly at 2-48 hours after CLP (P<0.05). After use of TGF-β1, TNF-α was further increased. CONCLUSION: During sepsis, TGF-β1 can decrease the monocyte TLR4 expression and NF-κB in liver tissue, but facilitate the formation of proinflammatory mediator TNF-α. This finding indicates that TGF-β1 may play a role in promoting inflammatory response during sepsis, but this regulation is not via direct regulation of monocyte TLR4 in peripheral blood.

AIM: To examine the inhibition of matrix metalloproteinase (MMP) activity by doxycycline (Doxy) and its effect on vascular smooth muscle cells (SMCs) proliferation, neointimal hyperplasia and vascular remodeling. METHODS: The model of rat common carotid artery injury was established by balloon-dilatation. Doxy was administered to the animals of treatment group at dose of 30 mg?kg -1?d -1. The activity of MMPs in the tissue of injured carotid arteries was measured by gelatin zymography. The thickness and area of neointimal, lumen area and the proliferation of SMCs were measured by histological and morphometric analysis. RESULTS: 1. After Doxy treatment, the activity of MMP-9 in the carotid arteries was reduced by 26.3% and 34.5% compared to that in rats without Doxy treatment at 24 hours and 3 days after balloon injury, respectively (P

Virtual experiments teaching has been characterized by openness, interactivity and re-source sharing. It efficiently improves the experiments teaching effects and promotes the teaching reform. At present the virtual experiment systems used by domestic universities can realize simulation of the ex-perimental principle, apparatus, object, operation and data. In the virtual experiment system students deepen the understanding of the experiments through foregrounding and the networked virtual experiment manage-ment effectively improves the effects of experiments teaching through behind-the-scenes action.

Objective To investigate the roles of phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway and its regulatory protein peroxisome proliferator-activated receptor γ (PPAR γ) and phosphatase and tension homologue deleted on chromosome 10 (PTEN) in regulating insulin sensitivity in rats with fetal growth restriction (FGR).Methods Sixteen pregnant rats were randomly divided into two groups including FGR and control groups on the 12th day of pregnancy (eight in each group).The FGR group was given low protein diet (8％ of casein) and restriction diet to establish the neonatal rat model of FGR.All maternal rats after delivery and newborn rats after weaning on 21 days after born were fed with normal diet.Each time blood samples were collected from eight newborn rats of each group to measure levels of fasting plasma glucose (FPG) and fasting insulin(FINS) at the time points of 21 days,two and four months after birth.Then insulin resistance index (IRI) and insulin sensitivity index (ISI) were calculated to evaluate insulin sensitivity.Expression of PI3K,AKT,PPAR γγ,PTEN and glucose transporters 4 (GLUT4) in skeletal muscle at mRNA and protein levels were measured at 21 days,two and four months after birth with real time fluorescence polymerase chain reaction and Western blot,respectively.Relationships between the expression of key molecules of PI3K/AKT signaling pathway and insulin sensitivity were analyzed.T-test,and Pearson's correlation analysis were used for statistical analysis.Results (1) The average birth weight of newborn rats in the FGR group was lower than that of the control group [(4.37± 0.69) vs (7.03±0.55) g,t=-20.75,P＜0.05].The incidence of FGR in the FGR group was 93.33％ (70/75).(2) Compared with normal offspring,those in the FGR group showed significantly increased FPG [two months after birth:(5.53± 0.58) vs (7.49 ± 0.38) mmol/L,t=8.08;four months afterbirth:(6.35±0.66) vs (8.94±0.90) mmol/L,t=6.58],FINS [two months afterbirth:(9.18±0.66) vs (14.67± 1.90) mU/L,t=7.71;four months after birth:(33.08±2.76) vs (56.33±2.81) mU/L,t=16.71] and IR1 (two months after birth:2.25±0.31 vs 4.90±0.81,t=8.63;four months after birth:9.30±0.90 vs 22.44±3.10,t=1 1.51),but decreased ISI (two months after birth:0.020 ± 0.002 vs 0.009± 0.001,t=-10.1 4;four months after birth:0.005±0.000 vs 0.002 ±0.000,t=-14.91) at two and four months after birth (all P＜0.05).(3) Compared with normal offspring,those in the FGR group showed decreased expression of PI3K (21 days after birth:0.082±0.028 vs 0.019±0.004,t=-6.29;two months after birth:0.020±0.003 vs 0.010±0.005,t=-4.78;four months after birth:0.014±0.004 vs 0.003±0.001,t=-7.87) and GLUT4 (21 days after birth:0.132±0.057 vs 0.041 ±0.019,t=-4.32;two months after birth:0.183±0.084 vs 0.069±0.017,t=-3.74;four months after birth:0.248±0.069 vs 0.113±0.040,t=-4.74) at mRNA level at 21 days,two and four months after birth (all P＜0.05).Compared with normal offspring,decreased expression of PPAR γ (two months after birth:0.028±0.002 vs 0.012±0.005,t=-3.70;four months after birth:0.030±0.008 vs 0.012±0.005,t=-3.80) and increased expression of PTEN (two months after birth:0.020±0.004 vs 0.045±0.014,t=5.09;four months after birth:0.023±0.007 vs 0.034±0.009,t=2.57) at mRNA level were observed in offspring of the FGR group at two and four months after birth (all P＜0.05).(4) Compared with normal offspring,expression of PI3K protein (21 days after birth:0.22±0.01 vs 0.17±0.02,t=-6.62;two months after birth:0.27±0.03 vs 0.16±0.02,t=-7.25;four months after birth:0.18±0.01 vs 0.09±0.02,t=-9.79) and GLUT4 protein (21 days after birth:0.21 ±0.01 vs 0.03±0.01,t=-27.29;two months after birth:0.10±0.01 vs 0.06t±0.01,t=-3.90;four months after birth:0.13 ±0.01 vs 0.08± 0.02,t=-8.10) decreased in offspring in the FGR group at 21 days,two and four months after birth (all P＜0.05).Compared with normal offspring,those in the FGR group showed decreased expression of PPAR γ protein (two months after birth:0.10 ± 0.01 vs 0.07± 0.01,t =-7.29;four months after birth:0.09±0.01 vs 0.08±0.01,t=-2.83),but increased expression of PTEN at protein level (two months after birth:0.10±0.01 vs 0.15±0.02,t=6.01;four months after birth:0.09±±0.01 vs 0.13±0.02,t=5.51) at two and four months after birth (all P＜0.05).(5) The IRI levels in offsprings in the FGR group were negatively correlated with the expression of PI3K,GLUT4 and PPAR γ at protein level (two months after birth:r=-0.90,-0.92 and-0.79;four months after birth:r=-0.92,-0.75 and-0.73,all P＜0.05),but positively correlated with the expression of PTEN at protein level (r=0.87 and 0.86,both P＜0.05) at two and four months after birth.Conclusions The abnormal expression of the key molecules of PI3K/AKT signaling pathway precedes the decrease of insulin sensitivity in newborn rats with FGR and the expression regulatory protein PPAR γ and PTEN are also changed,suggesting that these molecules may induce the impairment of insulin sensitivity in rats with FGR and be involved in the development of insulin resistance.

Objective To investigate the expression of microRNA-548ah (miR-548ah) and microRNA-4804 (miR-4804) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis B virus infection and their clinical significance.Methods PBMCs were collected from 29 patients with chronic hepatitis B (CHB),30 hepatitis B virus carriers (HBVC),26 inactive HBsAg carriers (IASC) and 28 healthy controls in Taizhou People's Hospital during September 2012 and August 2013.Expressions of miR-548ah and miR-4804 in PBMCs were detected by fluorescence real-time quantitative RT-PCR (qRT-PCR).Receiver operating characteristic (ROC) curve was used to evaluate the expression of miR-548ah and miR-4804 in distinguishing immune tolerance phase and clearance phase of HBV infection.Pearson correlation analysis was performed to assess the correlations of miRNAs expression with clinical markers alanine aminotrans ferase (ALT) and HBV DNA loads.Results There were significant differences in expressions of miR-548ah and miR-4804 in PBMCs between CHB,HBVC,IASC groups and control group (F =28.16 and 83.17,P ＜ 0.01).Compared with control group,miR-548ah was up-regulated in CHB group(P ＜ 0.01),and down-regulated in HBVC and IASC groups (P ＜ 0.01) ; miR-4804 was up-regulated in CHB group (P ＜ 0.01),down-regulated in HBVC group (P ＜ 0.01),while there was no significant difference between IASC group and control group in miR-4804 expressions(P ＞ 0.05).The areas under ROC curve (AUCs) of miR-548ah and miR4804 in differentiation of immune tolerance and immune clearance were 0.966 and 0.997,and the sensitivities and specificities were 89.7％,96.6％ and 99.6％,100.0％,respectively.No significant correlation was found between the expression of miR-548ah,miR-4804 and ALT,HBV DNA loads (r=0.14,0.18,-0.20 and-0.19,P＞0.05).Conclusion miR-548ah and miR-4804 may be involved in the immunopathogenesis of CHB,and their expression levels in PBMCs are helpful in differentiation of immune tolerance and immune clearance in HBV infection.

Objective Mining and exploring the positive experience of family caregivers of diabetic foot,aims to deepen the conceptual framework of positive aspects of caregiving and to provide a basis for interventions. Methods A qualitative study design was used guided by the conceptual framework of positive aspects of caregiving.Ten family caregivers of patients with diabetic foot were surveyed. The Colaizzi qualitative analysis method and the framework method were used to analysis the qualitative data. Results The caregivers′positive experience was described from 4 aspects:quality of caregiver/care receiver daily relationship, caregiver′s feeling of accomplishment, meaning of the caregiver′s role in daily life and the caregivers′gain. The caregivers′positive experience was influenced by the caregiver′s sense of self-efficacy and the enrichment events in daily life. Conclusions Family caregivers of diabetic foot get positive experiences in caregiving. The conceptual framework of positive aspects of caregiving explains the relationship between caregivers′positive experience and the determine factors, and it will be helpful to the designing and implementing of interventions. The positive experience should be considered when an intervention program will be conducted for supporting family caregivers.

Objective To study the influence of hydrogen sulfide (H2 S)on the mitochondrial function of pulmonary artery smooth muscle cells of the rats with high pulmonary blood flow pulmonary hypertension, and to clarify the function and its mechanism in the occurrence of high pulmonary blood flow pulmonary hypertension.Methods A total of 27 rats were randomly divided sham operation group,operation group,and operation+sodium hydrosulfide (NaHS)group (n=9).The pulmonary hypertension rat model was built by left lung resection.After fed for 35 d, the mean pulmonary artery pressure (mPAP),ratio of right ventricle/body weight (RV/BW),and ratio of right ventricle/(left ventricle+septum)[RV/(LV + S)]of the rats in three groups were measured.The plasma H2 S levels and the CSE activities in pulmonary artery tissue of the rats were detected;the activities of total mitochondrial ATP enzyme,superoxide dismutase (SOD),glutathione peroxidase (GSH-Px)and malondialdehyde (MDA)levels were determined;the ultrastructure of pulmonary artery smooth muscle mitochondria was observed through transmission electron microscope.Results Compared with sham operation group,the plasma H2 S level and the CSE activity in pulmonary artery tissue of the rats in operation group were decreased(P<0.01);the mitochondrial membrane of pulmonary tissue was swelling, and the mitochondrial activity was decreased (P<0.01);the mitochondrial ATP enzyme,SOD and GSH-Px activities were significantly decreased,and the MDA level was significantly increased (P<0.01).Compared with operation group,the H2S level in plasma and the activity of CSE in pulmonary tissue of the rats in operation + NaHS group were increased (P<0.01 );the mitochondrial membrane swelling was reduced, and the vitality was restored;the ATP enzyme, GSH-Px, and SOD level in pulmonary tissue were significantly increased (P<0.01),and the MDA level was significantly reduced (P<0.01).Conclusion H2 S can enhance the activities of mitochondrial ATP enzyme,GSH-Px,and SOD,and decrease the mitochondrial lipid peroxidation level,thus it plays a protective effect on rat pulmonary artery smooth muscle.