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

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.

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 level of foot care behavior and analyse related influencing factors in patients with high risk of type 2 diabetic foot (DF).Methods Apply the Chinese version of the Nottingham Assessment of Functional Footcare to 300 cases of patients with high risk of DF and take oneway ANOVA and multiple stepwise linear regression analysis on the general information and current foot condition.Results The score of foot care behavior was (45.58 ±7.83) scores.52.3 percents patients' scores were less than 45.Gender,previous education of diabetic foot care and history of diabetic foot ulcer were influencing factors of foot care behavior in patients with high risk of type 2 diabetic foot (t=3.196,3.392,2.462,P＜0.05).Conclusions The current level of foot care behavior was relatively low in patients with high risk of type 2 DF.It was also approved that female patients with high risk of DF who had received diabetic foot care education and suffered from diabetic foot ulcer before score much higher on foot care behavior.

Objective To explore the expression profiles and their clinical significance of microRNA (miRNA) molecules in different stages of chronic hepatitis B virus (HBV) infection.Methods The miRNA molecule expressions of 12 patients with chronic hepatitis B,12 chronic HBV carriers,12 inactive hepatitis B surface antigen (HBsAg) carriers,and 9 healthy controls were screened using miRNA microarray.The miRNA profiles were validated by the real time fluorescence quantitative polymerase chain reaction (PCR).The t-test was used for comparison between twogroups,whereas one-way ANOVA and SNK-q tests were used for multiple comparisons.Mann-Whitney and Kruskal Wallis H tests were used for comparison of two or more groups of data with skewed distribution.The receiver operation characteristic (ROC) curve was constructed to evaluate the diagnostic significance of miRNA molecules.Results Compared with the healthy controls,significant differences in the expression profiles of miRNA molecules were found in peripheral blood mononuclear cells of chronic HBV carriers (2 molecules up-regulated,and 18 down regulated) and chronic hepatitis B patients (33 molecules up-regulated and 19 down-regulated).No significant difference was found between inactive HBsAg carriers (2 molecules up regulated,and 3 down-regulated) and controls.The results of six miRNA molecules detected by real-time fluorescence quantitative PCR were basically consistent with the results detected by microarray.The area under ROC curve of the six miRNA molecules of hsa miR-4711-3p,hsa-miR-3191 5p,hsa-miR-5704-5p,hsa-miR 548ah-5p,hsa-miR-146a-5p and hsa-miR-29b-3p in distinguishing immune tolerance and clearance of chronic HBV infection were 0.994,0.984,0.967,1.000,1.000 and 0.996,respectively.Conclusions The different expression profiles of miRNA molecules could be used to distinguish the different stages of chronic HBV infection,and are closely related with the immune tolerance and activation in chronic HBV infection.

Objective To observe the effect of comprehensive nursing of traditional Chinese medicine and Western Medicine on children′s supracondylar humerus fracture of ulnar deviation on postoperative recovery. Methods From November 2015 to February 2016, 52 inpatients were divided into 2 groups by random digits table method after the external fixation removal surgery with 26 cases each. The observation group was intervened by integrated nursing and was given TCM rehabilitation guidance, while the control group was intervened by regular nursing and was given regular rehabilitation guidance. After 4 weeks, the therapeutic effect was evaluated according to Chinese Medicine Orthopedic Syndrome Diagnosis and their elbow joint function was evaluated by Flynn score. In 8 weeks of follow up, all the patients were valued by their quality of life. Results In the process of intervention, with 1 case fell off in the observation group, 3 cases fell off in the control group, so finally the observation group with 25 cases, the control group with 23 cases. Fifteen cases were cured, 7 cases were improved, 3 cases were not cured at 4 weeks after surgery in the observation group, 8, 6, 9 cases in the control group, and the difference was statistically significant (Z=-2.120, P=0.034). At 4 weeks after surgery, 10, 6, 6, 3 cases were excellent, good, medium and poor evaluated by Flynn score in the observation group, 5, 4, 5, 9 cases in the control group, and the difference was statistically significant (Z=-2.051, P=0.040). Incidence of postoperative complications was 8.0% (2/25) in the observation group, 30.4% (7/23) in the control group, and the difference was statistically significant (χ2=2.622, P=0.047). Physical, psychological, social and environmental fields of quality of life were 77.84 ± 12.15, 81.13 ± 15.72, 79.95 ± 20.68, 79.44 ± 11.57 in the observation group, 63.31 ± 15.86, 63.07 ± 12.63, 65.65 ± 18.51, 61.32 ± 17.24 in the control group, and the difference was statistically significant (t=2.516- 4.363, P<0.05). Conclusions Integrated nursing intervention with TCM rehabilitation guidance is a method which can effectively help in improving the elbow movement function and the life quality of children with supracondylar humerus fractures of ulnar deviation.