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

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

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.

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.

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 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 observe the antioxidative capability and the mRNA expression of sodium pump αl-subunit in kidney of hypothyroid rats by iodine deficiency and illuminate the pathogenesis of kidney damage.Methods Wistar rats were randomly divided into two groups as control group (NT) and hypothyroid group (HT).The rats were all fed with low-iodine diet derived from an endemic goiter area and drank deionized water containing different potassium iodide to duplicate hypothyriod animal models.We determined the morphometric parameters of kidney by routine histology method.The contents of malondialdehyde and free radical scavengers (GSH-PX and SOD) in kid-ney,as well as the activity of Na~+-K~+ ATPase were measured in two groups.The mRNA expression of sodium pump α1 subunit was determined by reverse transcription-polymerase chain reaction.Results Compared with that in the control group,in hypothyroid group (1) serum free T3,free T4,total T3 and total T4 were markedly lower.(2) mean glomerular area and volum diminished markedly.(3)the content of MDA and activity of PGx increased markedly,but the activity of SOD decreased significantly,as well as the one of Na~+-K~+-ATPase.(4)the mRNA expression level of sodium pump α1 subunit was lower.Conclusion In a hypothyroid state,the decrease of antiox-idative capability of kidney resulted in lipid peroxidative damage,atrophy of kidney,decreased activity of Na~+-K~+ -ATPase and degression of sodium pump α1 subunit mRNA expression.