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

AIM: To adopt the foam separation to get the best condition of the process about separation-purification sapindus-saponin. METHODS: The orthogonal experiments was used to analyse the results of the process. RESULTS: The best condition of the process was 2.5 g/L feed concentration,0.9 L/min gas flow rate,pH 4.8 and the temperature of 30 ℃.Under this condition,the yield,concentration ratio and purity of sapindus-saponin was 69.42%,2.48,and 67.78%,respectively. CONCLUSION: The process is very simple and practical,which provides a base for the application of natural sapindus-saponin.

Chromatography, Gas ; methods ; Dimethoate ; blood ; Humans ; Insecticides ; blood ; Methyl Parathion ; blood

Objective To investigate the electrophysiological characteristics of the ventralis intermedius nucleus (Vim) in order to find an easy and safe way to confirm the target in Vim-thalamotomy. Method In microelectrode-guided selective Vim-thalamotomy for 23 Parkinson's disease patients, the background activity, amplitude and discharge frequencies of Vim were compared with its surrounding structures. The response of kinesthetic neuron and tremor cell to microstimulation was also compared. Result There were differences in backgroud activity and discharge amplitude for Vim, ventralis lateralis nucleus (VL), ventralis caudalis nucleus (VC), and internal capsule. Based on the response to active or passive movement of contralateral limb tremor cells were divided into two subgroups, which were different in localization. Contralateral tremor showed different response when the two subgroups of tremor cells were mircrostimulated. Conclusion The anterior border of Vim was easily found by microrecording. Only by combining microstimulation with microrecording could the posterior border of Vim and its interior and lateral ordination of target were identified exactly and safely. Kinesthetic neurons and tremor cells which responded to the movement of contralateral limbs should be destroyed.

Objective To analysis the varying degrees of the M protein staining after immunofixation electrophoresis(IFE)and study its applications in clinical diagnosis.Methods 196 cases of clinical serum samples were tested by using IFE,we analyzed the positive electrophoretic bands of M protein and performed statistical analysis by using SPSS17.0.The M proteins were analyzed ret-rospectively.Results 103 patients were diagnosed with monoclonal gammopathy in 196 patients with positive M protein bands,in-cluding 96 cases of multiple myeloma(MM)and 7 cases of other monoclonal gammopathy;93 patients were non-monoclonal gam-mopathy.By analyzing the M band staining in different clinical groups,we found that M bands were mainly with dense and thick staining in monoclonal immunoglobulin group,the dense staining rate of MM was 90.6%,and the difference between MM and the other monoclonal gammopathy was not significant(P >0.05).In contrast,M bands were in light and narrow staining in non-mono-clonal immunoglobulin group,the rate of which was 25.8%,the difference between non-monoclonal immunoglobulin group and monoclonal immunoglobulin group was statistically significant(P <0.01).The proportion of allelic band in MM,other monoclonal gammopathy,non-monoclonal gammopathy were 39.6%,28.6% and 2.2% respectively,the differences were statistically significant (P <0.01).Conclusion The M band,accompanied by allelic band in IFE staining,is helpful in the diagnosis of monoclonal gam-mopathy,especially MM.The appearance of M protein provides early warning of monoclonal gammopathy.

Objective To observe the fluctuations of blood glucose in type 2 diabetic patients treated by insulin hypodermic injection in different sites.Methods 30 patients with type 2 diabetes were randomly divided into three groups:abdominal,edltoid region and lateral femoral subcutaneous injection group.After half a year of insulin therapy,dynamic glucose monitoring for consecutive(71±10)hours was conducted,and(847±80)results of glucose Level were obtained.Results The average glucose had no difference in three group,but diurnal maximum and maximum fluctuation amplitude in patients of abdominal subcutaneous injection group were significantly lower than the Other groups(P＜0.05).The time percentage of glycemic fluctuations within 3.6～10.0mmol/L and diurnal minimum was higher compared to other groups(P＜0.05).Conclusion The site choice of insulin subcutaneous injection can influence the fluctuations of blood glucose in type 2 diabetes mellitus and abdominal subcutaneous tissue was the most ideal of the three subcutaneous injection sites.

OBJECTIVETo discuss the mechanism of gambogenic acid (GNA) in inducing the apoptosis of melanoma B16 cells.

METHODThe inhibitory effect of GNA on the proliferation of B16 cells was measured by the methyl thiazolyl tetrazolium (MTT) assay. The effect of GNA on B16 cells was detected by the Hoechst 33258 staining. The transmission electron microscopy was used to observe the ultra-structure changes of B16 cells. The changes in PI3K, p-PI3K, Akt, p-Akt, p-mTOR, PTEN proteins were detected by the Western blotting to discuss the molecular mechanism of GNA in inducing the apoptosis of B16 cells.

RESULTGNA showed a significant inhibitory effect in the growth and proliferation of melanoma B16 cells. The cell viability remarkably decreased with the increase of GNA concentration and the extension of the action time. The results of the Hoechst 33258 staining showed that cells processed with GNA demonstrated apparent apoptotic characteristics. Under the transmission electron microscope, B16 cells, after being treated with GNA, showed obvious morphological changes of apoptosis. The Western blot showed a time-dependent reduction in the p-PI3K and p-Akt protein expressions, with no change in p-PI3K and p-Akt protein expression quantities. The p-mTOR protein expression decreased with the extension of time, where as the PTEN protein expression showed a time-dependent increase.

CONCLUSIONGNA could inhibit the proliferation of melanoma B16 cells and induce their apoptosis within certain time and concentration ranges. Its mechanism in inducing the cell apoptosis may be related to PI3K/Akt/mTOR signaling pathways.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Melanoma ; metabolism ; pathology ; ultrastructure ; Mice ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; metabolism ; Terpenes ; pharmacology ; Xanthenes ; Xanthones ; pharmacology

This article introduced the development and application effect appraisal of Microbial Engineering CAI courseware for bio-engineering specialization. The courseware focuses on knowledge system integrity, content-rich and gives prominence to the key points. Pictures, animation and video, and audio effects are also utilized appropriately to achieving stimulate students interest in learning and then improve teaching and learning performance. The courseware concentrates on core content of the course, such as fermentation parameters detection and automatic control, and fermentation equipments. The courseware was manufactured using the Powerpoint software. Animation was established with Flash 4 software and the scanning pattern was edited using Adobe photoshop. And chapters of the courseware were composed and administrated using Courseware Master Software. A two-year survey showed that 85% of students satisfied with this courseware.

Objective To study the effect of thyrotrophin releasing hormone (TRH) analogue, YM14673, on the brain edema and blood brain barrier after brain injury in rat.Methods The model of brain injury of rats was built by Feeney's methods. The Evans blue solution had been injected i.v. into the rats before the models were made. The rats were divided into four groups: normal, treated with saline, treated with YM14673(Ⅰ:0.1mg/kg and Ⅱ:1mg/kg). The water content in brain was measured 24 h after brain injured. The concentration of Evans blue in brain tissue and blood was measured with fluorometry.Results The rats treated with saline after traumatic injury showed significantly high water content compared with normal group(P<0.01)and the water content of the left hemisphere, which was hit straightly, was higher significantly than that of the right global(P<0.01). The brain water content decreased in the rats treated with YM14673 in all global(P<0.05).There was no significant different between the treated group Ⅰ and groupⅡ.After brain injury, the concentration of Evans blue in brain tissue showed a higher level contrasted with normal group. YM14673 did not influence the concentration of Evans blue in brain tissue. Conclusion YM14673 can decrease the brain edema after brain injury but it cannot decrease the permeability of blood brain barrier.

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.