Objective:To compare the human ??T lymphocytes expanded with HSP 70BCG and HSP 70 m-peptide complexes with those expanded with solid phase antibody in vitro.Methods:PBMCs were cultured with anti-TCR ?? antibody?HSP 70BCG and HSP 70 m -peptide complexes.The total cell number was counted.The flow cytometer was used to analyze the lymphocytes phenotypes and subtypes.RT-PCR was used to analyze the level of V? mRNA.The cytotoxitic activity of ??T cells against Daudi was determined using MTT calorimetric assay.Results:After 14 days of culture,in the antibody culture system,the total cell number increased about 30-40 fold,and the ratio of ??T cell reached to 86.3%;In the HSP 70BCG culture system,the total cell number increased about 7-8 fold,and the ratio of ??T cell reached to 71.23%;In the HSP 70-peptide complexes culture system,the total cell number increased about 4-5 fold,and the ratio of ??T cells reached to 27.26%.The antibody and HSP 70BCG activated ??T cells possessed a whole repertoire and mainly expressed V?9/V?2 subset and exhibited a strong cytotoxicity against Daudi.Conclusion:Anti-TCR ?? antibody and HSP 70BCG were able to proliferate purer ??T cells.The cells had a whole repertoire,and exhibited strong cytotoxicity against Daudi.

Objective To investigate the value of three-dimension (3D) ultrasound in identification of benign or malignant gastric ulceration. Methods A total of 88 patients with gastric ulcer were collected in Tianjin Nankai Hospital from March 2014 to May 2015, including 36 malignant cases and 52 benign cases confirmed by pathological results. Results of 2D ultrasound, 3D ultrasound and gastroscope diagnosis were compared, and the diagnostic values of the three methods were evaluated by the gold standard of pathologic results. Receiver operating characteristic (ROC) curves were plotted for the main measurement indicators (ulcer surface width, ulcer surface depth and thickness of stomach wall) of 3D ultrasound. The best cut-off value was determined. Results The positive rate of malignant gastric ulcer diagnosed by 3D ultrasound was significantly higher than that of 2D ultrasound (P=0.002), but there was no significant difference between 3D ultrasound and gastroscope detection (P=0.453). The diagnostic sensitivities of malignant gastric ulcer were 86.11%, 58.33%and 91.67%for 3D ultrasound, 2D ultrasound and gastroscope detection respectively. The diagnostic specificities were 100%, 100%and 96.15% respectively. The areas under the ROC curves measured by 3D ultrasound were 0.750, 0.940 and 0.977 for ulcer surface width, ulcer surface depth and thickness of stomach wall. And the best cut-off values of the three indicators were 16.55 mm, 8.05 mm and 9.90 mm. Conclusion The 3D ultrasound can show the form and structure of gastric ulcer more clearly and intuitively, which is valuable for the differential diagnosis of benign gastric ulcer and malignant gastric ulcer, and may be used for screening malignant gastric ulcer.

The zinc finger antiviral protein (ZAP) specifically inhibits the replication of Moloney murine leukemia virus (MLV) and Sindbis virus (SIN), but has very modest inhibitory effect on HIV. Previous studies suggest that ZAP directly binds to the viral RNA and recruit the RNA degradation machinery to degrade the target RNA. The HIV-1 Tat and Rev are regulatory proteins which bind to HIV RNA. Tat and Rev were fused with ZAP in various manners. Two fusion proteins, ZAP-Tat and ZAP-Rev were found to be able to inhibit the expression HIV-1 vector.

Objective To study the changes and the role of MCP -1,IL -8,VEGF,NO,NOS in the T2DM patients with different types of CHD.Methods According to the result of coronary arteriongraphy and clinical symp-toms,and the diagnostic code of T2DM established by Chinese Medical Association diabetology branch in 2007, 126 patients of T2DMwith CHD were chosen and divided into two groups:ACS +T2DM group (A group,74 cases) and SAP +T2DMgroup (B group,52 cases),in addition,50 healthy people were chosen as control group.The levels of MCP -1,IL -8,VEGF were measured by the method of ELISA.The level of NO was measured by the method of nitrate reductase and NOS activity was measured by the method of spectrophotometer.Then,the results were analyzed. Results The levels of MCP -1 and IL -8 in A group and B group were[(115.98 ±39.57)pg/mL,(98.76 ± 31.55)pg/mL],[(131.22 ±42.83)pg/mL,(115.75 ±40.37)pg/mL],which were all higher than those in group C [(75.63 ±23.69)pg/mL,(68.53 ±37.85)pg/mL,t =4.12,2.26,3.78,2.21,all P <0.05)],but the VEGF [(167.87 ±54.98)pg/mL,(128.38 ±36.99)pg/mL)],NO[(46.89 ±12.92)μmol/L,(51.66 ±12.49)μmol/L)]and NOS [(39.04 ±5.19)U /mL,(40.56 ±7.03)U /mL)]were lower than those in C group [(90.21 ± 32.06)pg/mL,(64.05 ±13.58)μmol/L,(47.82 ±5.93)U /mL;t =3.05,3.17,2.43,2.79,2.49,2.25,all P <0.05].The MCP -1,IL -8 levels in A group were higher than those in B group(t =3.13,2.89,all P <0.05),but the level of VEGF and NO were lower than that in B group(t =3.04,2.95,all P <0.05),NOS in A group was lower than that in B group,but there was no statistically significant difference between the two groups(t =1.06,P >0.05). MCP -1 was positively correlated with Il -8,VEGF (r =0.35,0.33,all P <0.01),and it had negative correlation with NO (r =-0.24,P <0.05).Conclusion Inflammatory factor and oxidative stress both participate in the T2DM with different types of CHD,it relates with the degree of CHD.

Objective: To discuss the action mechanisms of Takayasus arteritis (brachiocephalic artery type) of acupuncture. Methods: We applied acupuncture therapy with the principle of warming Yang and supplementing Qi, removing obstruction in the meridians and recovering pulse, to treat patients with takayasus arteritis (brachiocephalic artery type). Before and after treatment, we detected the TCD (Transcranial Doppler) changes of the average velocity of blood flow (AVBF) and arterial pulsatility index (PI) of related arteris. Results: After treatment,acupuncture and moxibustion can regulate the abnormal blood flow rate in endocranial correlated arteries by two sides, in the AVBF while, elevate the elasticity of endocranial vessels.Conclusion: Acupuncture and moxibustion can regulate cerebrovascular function of patients with takayasus arteritis (brachiocephalic artery type), increase the perfusion of cerebral blood flow, and adjust abnormal state of endocranial hemodynamics. This is possibly the important action mechanisms acupuncture in treating this disorder.

Small interfering RNAs (siRNAs) can efficiently inhibit gene expression by sequence-specific RNA interference (RNAi). A common 5' leader sequence exists in the genomic RNA and all subgenomic RNAs of SARS-CoV, and is well conserved among various SARS-CoV strains, thus providing a preferable target for RNAi of SARS-CoV replication. Here efficient depletion of the SARS-CoV mRNAs by either a synthetic siRNA or DNA vector-derived short hairpin RNAs (shRNAs) targeting the leader sequence in mammalian cell lines were reported. The siRNA or shRNAs efficiently suppressed the expression of an EGFP reporter gene which contains the leader sequence at the 5' end. Both the siRNA and shRNAs efficiently knocked down the levels of leader-containing transcripts of three SARS-CoV genes encoding the spike protein, membrane protein and nucleocapsid protein were demonstrated. The results suggest that RNAi targeting the leader sequence is a potential efficient strategy for anti-SARS-CoV therapy.

ObjectiveTo study the association of SNP276 in adiponectin gene with impaired glucose regulation (IGR) complicating with the different component numbers of metabolism syndrome (MS) in population of IGR in Han people of Shanxi region.MethodsThe study population consisted of 256 subjects with IGR which was composed of 123 subjects with component numbers of MS ＜2 (IGRA) and 133subjects with component numbers of MS≥2 (IGRB),and 128 subjects with normal healthy (Normal) who were Chinese Hans residents and in Shanxi province.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to test the adiponectin SNP276G/T polymorphism.ResultsThe distributions of genotypes and alleles of SNP276 both displayed significant difference among the three groups ( x2 =16.893,P =0.002 ; x2 =18.149,P =0.000).In the IGRA,the SNP276 non-TT ( GT + GG) genotype was no difference which increased risk of complicating with the different component numbers of MS( P =0.781,P =0.809).In IGRB,the SNP276 non-TT ( GT + GG) genotype was associated with increased risk of complicating with the different component numbers of MS,and after age and sex-adjusted,there was significant difference ( P =0.007,P =0.007).ConclusionsSNP276 in APM1 increased the risk of complicating with the components of MS in population with impaired glucose regulation in Han people of Shanxi region.

Hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs) is a key component of the endosomal sorting complexes required for transport and has been demonstrated to play a regulatory role in endocytosis/exocytosis and the accumulation of internal vesicles in multivesicular bodies. Citron kinase is a Ser/The kinase that we previously reported to enhance human immunodeficiency virus type 1 (HIV-1) virion production. However, the relationship between Hrs and citron kinase in HIV-1 production remains elusive. Here, we report that Hrs interacts with citron kinase via its FYVE domain. Overexpression of Hrs or the FYVE domain resulted in a significant decrease in HIV-1 virion production. Depletion of Hrs by RNA interference in HEK293T cells increased HIV-1 virion production and enhanced the activity of citron kinase. These data suggest that Hrs inhibits HIV-1 production by inhibiting citron kinase-mediated exocytosis.
Down-Regulation ; Endosomal Sorting Complexes Required for Transport ; genetics ; metabolism ; Endosomes ; metabolism ; Exocytosis ; Gene Expression ; Gene Silencing ; drug effects ; HEK293 Cells ; HIV Infections ; genetics ; metabolism ; virology ; HIV-1 ; drug effects ; genetics ; growth & development ; Humans ; Immunoprecipitation ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Microscopy, Fluorescence ; Phosphoproteins ; genetics ; metabolism ; Plasmids ; Protein Binding ; drug effects ; genetics ; Protein Interaction Domains and Motifs ; Protein Structure, Tertiary ; Protein Transport ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; pharmacology ; Transfection ; Virion ; drug effects ; genetics ; growth & development ; Virus Release ; drug effects ; Virus Replication ; drug effects

The zinc-finger antiviral protein (ZAP) is a host factor that specifically inhibits the replication of certain viruses, including murine leukemia virus, Sindbis virus and Ebola virus, by targeting the viral mRNAs for degradation. ZAP directly binds to the target viral mRNA and recruits the cellular RNA degradation machinery to degrade the RNA. No significant sequence similarity or obvious common motifs have been found in the so far identified target viral mRNAs. The minimum length of the target sequence is about 500 nt long. Short workable ZAP-binding RNAs should facilitate further studies on the ZAP-RNA interaction and characterization of such RNAs may provide some insights into the underlying mechanism. In this study, we used the SELEX method to isolate ZAP-binding RNA aptamers. After 21 rounds of selection, ZAP-binding aptamers were isolated. Sequence analysis revealed that they are G-rich RNAs with predicted stem-loop structures containing conserved "GGGUGG" and "GAGGG" motifs in the loop region. Insertion of the aptamer sequence into a luciferase reporter failed to render the reporter sensitive to ZAP. However, overexpression of the aptamers modestly but significantly reduced ZAP's antiviral activity. Substitution of the conserved motifs of the aptamers significantly impaired their ZAP-binding ability and ZAP-antagonizing activity, suggesting that the RNA sequence is important for specific interaction between ZAP and the target RNA. The aptamers identified in this report should provide useful tools to further investigate the details of the interaction between ZAP and the target RNAs.
Aptamers, Nucleotide ; chemistry ; genetics ; metabolism ; Base Sequence ; Carrier Proteins ; antagonists & inhibitors ; metabolism ; Genes, Reporter ; HEK293 Cells ; Humans ; Leukemia Virus, Murine ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Protein Binding ; RNA ; chemistry ; metabolism ; RNA, Viral ; genetics ; Response Elements ; SELEX Aptamer Technique

HIV-1 utilizes cellular factors for efficient replication. The viral RNA is different from cellular mRNAs in many aspects, and is prone to attacks by cellular RNA quality control systems. To establish effective infection, the virus has evolved multiple mechanisms to protect its RNA. Here, we show that expression of the Y-box binding protein 1 (YB-1) enhanced the production of HIV-1. Downregulation of endogenous YB-1 in producer cells decreased viral production. YB-1 increased viral protein expression by stabilizing HIV-1 RNAs. The stem loop 2 in the HIV-1 RNA packaging signal was mapped to be the YB-1-responsive element. Taken together, these results indicate that YB-1 stabilizes HIV-1 genomic RNA and thereby enhances HIV-1 gene expression and viral production.
5' Untranslated Regions ; Chromosome Mapping ; Down-Regulation ; HEK293 Cells ; HIV-1 ; genetics ; metabolism ; Humans ; Inverted Repeat Sequences ; Protein Binding ; RNA Interference ; RNA, Small Interfering ; metabolism ; RNA, Viral ; metabolism ; Virus Replication ; Y-Box-Binding Protein 1 ; antagonists & inhibitors ; genetics ; metabolism