Objective:To study the changes of the NKT like cells after HIV infected. Methods:We collected peripheral blood from 47 untreated HIV infected individuals and 31 healthy controls,and analyzed the expression of Annexin-V,Ki-67,HLA-DR and other surface molecules in NKT like cells by flow cytometry. Results:The NKT like cell percentage of untreated HIV infected group was (3. 03±1. 61)%,the NKT like cell percentage of normal control group was (8. 30±7. 42)%,the percentage of NKT like cells in HIV infected individuals was significantly lower than the healthy controls ( P<0. 05 );the NKT like cell HLA-DR expression of untreated HIV infected group and normal control group were (5. 40±4. 10)% and (0. 89±0. 83)%,the NKT like cell Annexin-V expression of untreated HIV infected group and normal control group were (30. 21±13. 15)% and (5. 40±8. 05)% ,and the activation and apoptosis of NKT like cells was significantly higher after HIV infection compared with health individuals (P<0. 001,P<0. 01),the degree of activation was negatively correlated with CD4 count (r=-0. 885 7,P<0. 05);and the NKT like cell Ki-67 expression of untreated HIV infected group and normal control group were (11. 15±4. 76)% and (27. 63±18. 31)%,the proliferation ability was significantly lower after HIV infection compared with healthy controls(P<0. 05). Conclusion:HIV infection can significantly reduce the number of NKT like cells and its ability to proliferate,and increase its ability to activation and apoptosis.

Objective:To better understand the changes of the NKT like cells after HIV infection and HAART treatment.Methods: Peripheral blood from HIV-infected individuals, HAART-treatment AIDS patients and healthy controls were collected, the expression of CD57 and the proliferation ability of NKT like cells before and after HAART were analyzed by flow cytometry.Results:We found that the percentage of NKT like cells before HAART was significantly lower than the healthy controls ( P<0.01 ) , and recovered after HAART treatment ( P<0.05 );the aging of NKT like cells was significantly higher before HAART compared with health individuals (P<0.01),and recovered after HAART treatment(P<0.05)the proliferation was significantly lower in vitro before HAART compared with healthy controls,and partial recovered after HAART.Conclusion: After HAART treatment,the number of NKT like cells,CD57 expression and the proliferation ability of HIV infected patients were restored.

ObjectiveTo screen mimetic HIV-1 neutralizing epitopes from plasma with high level neutralizing antibody,and to provide useful information for further study of the interaction between antigen and antibody.MethodsIn order to gain neutralizing antibody recognized mimotopes, we detected neutralizing antibodieslevelsof 11HIV-1infectedslowprogressorsbyPBMC-basedneutralization assays.High-titer HIV-neutralizing antibodies from plasma of SPs was used as the ligand for biopanning by phage-displayed random peptide library.Positive phage clones was evaluated by ELISA,sequenced,and analyzed for homology to HIV-1 env by local BLAST to deduce the neutralizing peptide.ResultsTwenty-two clones were obtained consistent with requirement through three rounds biopanning.After comparison analysis,twelve clones include C8 were obtained as mimotopes of neutralizing antibody,C40 located in gp41Ⅱ cluster with the highest titer by inhibition ratio may be as neutralizing epitope.Conclusion By the use of IgG antibodies from SPs to screen the phage random polypeptide library,one can acquire multiple phage mimetic peptides of HIV related antigen epitope.(Chin J Lab Med,2012,35:838-842 )

Objective To examine the expression of TLR7/8 in monocytes purified from HIV-1 infected individuals and to study its association with disease progression. Methods Sixty-three HIV-1 infected individuals and 18 normal controls were enrolled. Monocytes were purified by MACS system and RNA of them was extracted by RNA mini kit of QIAGEN company. TLR7/8 expression was tested by real-time RT-PCR with ABI7500. Results It was found that the expression of TLR7 was strongly correlated with absolute CD4 count (r =0.614, P＜0.01) , so was TLR8 (r =0.419, P＜0.01). The expression of TLR7 in slow progressor (SP) group was higher than that in HIV-1 infected patients group, AIDS patients group and normal group (P ＜ 0. 05 ) . HIV group and normal group were strongly higher than AIDS group (P ＜ 0. 05). It was no significant differentiation of expression of TLR7 between HIV infection group and normal control group. The expression of TLR8 in SP group and normal group were significantly higher than that in AIDS group (P ＜ 0. 05). The expression of TLR8 was no singnificantly difference between SP group and HIV group or normal control group, so was it between HIV group or normal control group and AIDS group. Conclusion The expression of TLR7/8 in monocytes from HIV-1 infected patients significantly correlated with disease progression.

Objective To investigate the killer cell lg-like receptors (KIR) gene frequency of HIV-1 infected slow progressors(SP) and typical progressors(TP), and to analyze the interaction between KIR alleles and the progression of HIV-1 infection in Chinese population. Methods Eighty-one HIV-1 posi-tive individuals including 43 SPs and 38 TPs were recruited. Carriage of KIR genes was assessed using poly-merase chain reaction sequence-specific primers (PCR-SSP) assays. Results KIR2DS3 gene frequency was significantly lower in SP group (3.6%) than that in TP group (14.2%), P =0. 018 ,OR =0. 210,95% CI =0.053-0.833. The number of activating KIR genes was less in SP group than that in TP group, but was not significant (P = 0. 208). Conclusion Lower KIR2DS3 gene frequency may potentially be associated with slower progression to AIDS in Chinese population.

Objective To investigate the association between APOBEC3G mRNA levels in peripheral blood mononuclear cells(PBMCs)of HIV/AIDS patients and disease progression in Henan province.Methods Real-time PCR was used to detect the mRNA levels of APOBEC3G in PBMCs of HIV/AIDS patients at difierent disease progression stage.Flow cytometry and automated viral load analyzer were used to count CD4+ T cells and plasma HIV viral loads.Results The mRNA levels of APOBEC3G in HIV/AIDS patients were lower than in HIV-negative controls(t=4.887,P＜0.01),and APOBEC3G levels were significantly higher in slow development group than those in HIV and AIDS groups(P＜0.05).The levels of APOBEC3G mRNA correlated positively with CD4+ T cell counts(R2=0.190,P=0.002)and negatively with HIV-1 viral loads(R2=0.094.P=0.038).Conclusions The APOBEC3G mRNA levels in PBMC of HIV infected individuals are associated with HIV disease pmgression. Higher mRNA expression levels of APOBEC3G may be one of the protective factors which can play important role in delaying disease progression.

Objective To study the B lymphocytes counts and the expression of TLR9 mRNA on B lymphocytes in peripheral blood from Chinese HIV-infected patients.Methods The cells from peripheral blood were stained with antibodies labeled with fluorescence and B lymphocytes were counted with flow cytometry.Using the method of magnetic activated cell sorting and real-time PCR,the expression of TLR9 mRNA was measured.Results The B lymphocytes counts in HIV/AIDS patients was significantly lower than that of healthy controls(P ＜0.01).The B lymphocytes counts in HIV/AIDS patients positively correlated with the CD4 +T cells counts(r =0.534,P = 0.006).The expression of TLR9 mRNA on B lymphocytes in HIV/AIDS patients was significantly lower than that of healthy controls(P =0.023),and positively correlated with the CD4 + T cells counts(r = 0.390,P = 0.040).Conclusion The B lymphocytes counts and the expression of TLR9 mRNA on B lymphocytes in Chinese HIV/AIDS patients were decreased due to HIV infection,which may correlate with disease progression.

Objective To study the amino acid mutations in neutralizing antibody 2F5 and 4E10 conserved epitopes ELDKWA and NWFDIT of HIV-1 membrane proximal external region(MPER)in 92 HIV-infected individuals and AIDS patients in China,and to provide a basis for the neutralizing antobodies immunotherapy and a design of vaccines. Methods Nest-PCR methods were used to amplity genes of the HIV-1 env gp41 region.The amplified fragments were sequenced by double-deoxygen terminal method and translated into amino acids for analysis.The mutations of 2F5 and 4E10 neutralizing epitopes were identified by comparison with the epitopes reference data in HIV-1 Sequence Database.Results There were mutations on both 2F5 and 4E10 neutralizing epitopes.2F5 conserved neutralizing epitopes major mutations tocused on E662A(14.1%),K665S(17.4%),A667K(16.3%),and 4E10 conserved neutralizing epltopes major mutations included N671S(13.0%),D674S(3.3%),T676S(16.3%).The mutation rates of 2F5 and 4E10 epitopes were significanfly different between CRF_B'C-clade and B'-clade(P<0.05).The mutata rates of CRF_B'C-clade were higher than that of CRFOI_AE-clade in 2F5 epitopes(P<0.05).The mutation rates of B'-clade in 4E10 eiptopes showed significant difference in slow progressors,HIV-infected individuals and AIDS patients,respectively(P<0.05).Conclusion The HIV-1 patients in China are demonstrated diversified mutations in 2F5 and 4E10 neutralizing epitopes.The mutation degrees of amlno acids in conserved neutralizing epitopes are different in different subtypes.There may be a correlation between neutralizing epitopes mutations of 4E10 with disease progression.

Objective To investigate the clinical features,complications,treatment and prognosis of patients with Pcutz-Jeghers syndrome.Methods The clinical data of thirty-one cases of Pcutz-Jeghers syndrome treated in the First Hospital of Shanxi Medical University from January 1998 to December 2016 was analyzed retrospectively.Results There were11 cases with definite family history(35%,11/31).There were 21 cases of mucocutaneous pigmentation(67.7%,21/31),31 cases of gastrointestinal polyps(100%,31/31), including 8 cases of canceration(25.0%,8/31),24 cases of digestive tract bleeding(77.4%,24/31),20 cases of intussusception(64%,20/31),and 1 case of intestinal perforation(3.2%,1/31).6 cases were treated with endoscopic high frequency electrocoagulation,12 cases received polypectomy,17 cases underwent resection of intestinal tract.3 patients with intractable intussusception underwent plication of small intestine,and 4 patients with colon cancer underwent total colectomy combined with lymph node dissection.Conclusion Peutz-Jeghers syndrome often induce intractable intestinal obstruction and gastrointestinal tumor,resection of intestinal lesions should be favorable.For small intractable intussusception,the plication of small intestine has certain value; for its potential malignancy,it should be strictly monitored and actively treated.Gene therapy and drug therapy will guide for the future development.

Objective:To study the relationships between neutralizing antibody response against autologous virus and disease progression of HIV-1 B'/C infected individuals in China.Methods:Twenty-four primary HIV-1 isolates were incubated with autologous plasma collected either freshly or at approximately six months intervals thereafter.Normal human peripheral blood mononuclear cells were incubated with the virus-serum mixtures for 7 days and then the production of p24 antigen was measured.The neutralizing titer of a particular plasma and virus was defined as the reciprocal of the highest dilution giving a 50% reduction in p24 Ag compared with NHP control wells.More than 1∶8 were considered significant and were scored as positive.Results:In neutralizing antibody(Nabs) response against contemporaneous virus,Nabs were produced in all slow progressors(SP) individuals,while only four in 21 of HIV group had.There was statistically significance of the neutralizing antibody titers between SP and HIV.When plasma samples of six months later were tested for their ability to neutralize autologous virus,all of SPs had higher neutralizing antibody titers and the titers of neutralizing antibody in HIV group had increased in different rate.Among the twenty-one individuals of HIV group,12 of these individuals had neutralizing antibody response against autologous virus and other 9 of these individuals had not.NAb titers of SP in six months later plasma were higher than those of HIV.There was a negative correlation between the generation of the neutralizing titer against autologous virus and the plasma HIV RNA level in HIV-1 B'/C infected individuals(including SP,HIV).Conclusion:Neutralizing antibody against autologous viruses in HIV-1 B'/C infected SP is higher than those of HIV group,suggesting that neutralizing antibodies play a vital role in delaying disease progression in these individuals.