The Study On Mitochondrial DNA Polymorphism Of HIV/AIDS Patients And Effect Of Zhen Qi Fuzheng Granule On Immune Reconsititution Of Patients

AIDS(Acquired immunodeficiency syndrome),is a serious infectious disease caused by HIV(human immunodeficiency virus)infection,that mainly attacked CD4 positive immune cells,resulting in progressive damage to the immune function of the human body,prone to various opportunistic infections and tumors.Currently,the most effective treatment for HIV infection is the combination of a variety of an-tiretroviral drugs,that is,HAART(Highly active antiretroviral therapy),Even so,a considerable percentage of patients do not receive effective immune reconstitu-tion.Previous studies have found that mitochondrial DNA(mtDNA)polymorphism may have different effects on the disease progression of HIV infected individuals,as well as the efficacy and side effects of antiretroviral drugs,At the same time,as a useful supplement to HAART,Chinese medicine promotes the immune reconstitution of HIV/AIDS patients through "helping the healthy qi,regulating yin and Yang",and has achieved fruitful work in the prevention and control of aids.We attempted to ana-lyze the variation of base of mtDNA hypervariable region I and Therapeutic effect of ZhenQi Fuzheng granule combined with HAART on HIV/AIDS patients,in order to provide more individualized treatment strategies for HIV/AIDS patients from the molecular biology perspective,and to investigate the therapeutic effect of "Fuzheng"Chinese medicine on HIV infection.Part ? The study on mitochondrial DNA polymorphism of HIV/AIDS pa-tientsObjective:The base sequence of mtDNA hypervariable region ? in peripheral blood leu-kocytes of HIV/AIDS patients and healthy subjects was analyzed,To investigate the change of single nucleotide polymorphism in mtDNA hypervariable region ? in HIV/AIDS patients.Method:1.110 HIV/AIDS patients were selected as the case group,and 110 healthy subjects were taken as control group,the whole blood genome was extracted and sequences including mtDNA hypervariable region ? were amplified by PCR.2.PCR products were identified by agarose gel electrophoresis and purified by cen-trifugal adsorption column and were bidirectional sequencd,Compare the sequence information with the corrected standard Cambridge sequence,?2 test was used to analyze the variation of base by SPSS13.0 Software.Result:1.In the mtDNA hypervariable region ? in the case group,the average nucleotide mutation rate was 1.03%;However,that is 0.64%in the control group.The average base mutation rate in case group was significantly higher than the control group,there was statistical significance(p<0.01).2.In the case group,there were more than 20%mutation rates at 16129,16182,16183,16189,16223 and 16362 site,Compared with the control group,there was an upward trend,but there was no statistical difference.Conclusion:1.There was an increase in base mean mutation rate in the mtDNA hypervaria-ble region ? of HIV/AIDS patients.2.There were upward trend of site mutation rate of 16129,16182,16183,16189,16223,16362 in the mtDNA hypervariable region ? of HIV/AIDS patients,but further research was needed.Part ? The study on effect of Zhen Qi Fuzheng granule on immune reconsti-tution of HIV/AIDS patients and retrospective analysis of mitochondrial DNA polymorphism in patientsObjective:Zhen Qi Fuzheng granule were added to the HIV/AIDS patients for HAART,af-ter 3,6 months,investigated the difference of CD4+T cell count,effective rate of im-mune reconstitution,routine blood test,biochemical indexes between case group and control group.Retrospective analysis of the variation of base region of mtDNA hy-pervariable ? region was done in patients with different immune reconstitution effects.Method:1.In the case group of part ?,63 cases of HIV/AIDS patients matching selected condition treated by ZhenQi Fuzheng granule combined with HAART were selected as Chinese medicine+HAART group,75 other patients with HIV/AIDS treated with HAART alone served as control groud.2.The blood CD4+T cell count and routine blood test,biochemical indexes of two groups were detected before and after treatment in 3 and 6 months,variance analysis of repeated measurements,t-test and ?2-test were used to analyze the differ-ence of CD4+T cell count,effective rate of immune reconstitution,routine blood test,Biochemical indexes between the two groups through spss13.0 software.3.According to the variation of base in the first part of the study,the base muta-tion of mtDNA hypervariable region I Zhen Qi Fuzheng granule in +HAART group was analyzed through ?2-test.Result:1.Not stratified.The blood CD4+ T cells count in the two groups of patients af-ter 6 months treatment:The Chinese medicine+HAART group was 437.36+154.24;the control group was 351.04+123.98,P=0.019;The effective rate of immune recon-stitution:the Chinese medicine+HAART group was 71.2%,the control group was 52.9%,P=0.033.2.Stratified by 0?CD4+T cell count<200/?l.The blood CD4+T cells count in the two groups of patients after 6 months treatment:The Chinese medicine+HAART group was 437.3±154.24;the control group was 303.74±150.67,P=0.013;The effec-tive rate of immune reconstitution:the Chinese medicine group was 77.8%,the con-trol group was44.4%,P=0.027.3.Stratified by 200/?l?CD4+ T cell count<400/?l.The blood CD4+T cells count in the two groups of patients after 6 months treatment:The Chinese medi-cine+HAART group was 543.37±210.24;the control group was 467.53±130.41,P=0.023;The effective rate of immune reconstitution:the Chinese medicine group was 63.6%,the control group was 51.4%,P=0.30.4.Blood white blood cell(WBC)and hemoglobin(HB)index in two groups af-ter 6 months treatment:The control group:WBC:before treatment,5.15±2.03;After treatment,4.13±1.95*;P=0.034.Hb:before treatment,144.80±14.84;After treatment,123.84±14.12*;P=0.017.The Chinese medicine group:WBC:before treatment,4.55±1.06;After treatment,5.03±1.42;P=0.190.Hb:before treatment,136.30±25.46;After treatment,140.74±34.33;P=0.410.5.The rate of variation has an increasing tendency in 16129 and 16189 sites of mtDNA hypervariable region I in in the patients with poor immune reconstitution treated by Zhen Qi Fuzheng granule+HAART.Conclusion:1.After 6 months treatment,the CD4+T cell count and the effective rate of im-mune reconstitution in traditional Chinese+HAART medicine group were higher than that in control group.2.For HIV/AIDS patients(CD4+T cell count<200/?l),ZhenQi fuzheng gran-ule+HAART may have more advantages to immune reconstitution of patients.3.ZhenQi Fuzheng granule probably have protective and elevatory effect in WBC and Hb of HIV/AIDS patients undergoing HAART treatment.4.The sites of mtDNA hypervariable region I,such as 16129 and 16189,may be related to the immune reconstitution efficiency in HIV/AIDS patients.