2.Subtype and characterization of gag gene of HIV-1 strains prevalent in some areas of China
Li-Li, CHEN ; Yong-Jian, LIU ; Han-Ping, LI ; Zuo-Yi, BAO ; Dao-Min, ZHUANG ; Si-Yang, LIU ; Zheng, WANG ; Zhang-Li, LAI ; Lin, LI ; Jing-Yun, LI
Bulletin of The Academy of Military Medical Sciences 2009;33(6):505-508
Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.
3.Role of microRNAs in endothelial function.
Xian-bing CHEN ; Si-dao ZHENG ; Hong-jin WU
Chinese Medical Journal 2013;126(9):1779-1786
4.Roles and mechanisms of ginseng in protecting heart.
Si-Dao ZHENG ; Hong-Jin WU ; De-Lin WU
Chinese journal of integrative medicine 2012;18(7):548-555
Ginseng, the root of Panax ginseng C. A. Mayer, has long been used clinically in China to treat various diseases. Multiple effects of ginseng, such as antitumor, antiinflammatory, antiallergic, antioxidative, antidiabetic and antihypertensive have been confirmed by modern medicine. Recently, the clinical utilization of ginseng to treat heart diseases has increased dramatically. The roles of ginseng in protecting heart are foci for research in modern medical science and have been partially demonstrated, and the mechanisms of protection against coronary artery disease, cardiac hypertrophy, heart failure, cardiac energy metabolism, cardiac contractility, and arrhythmia are being uncovered progressively. However, more studies are needed to elucidate the complex mechanisms by which ginseng protects heart. All such studies will provide evidence of ginseng's clinical application, international promotion, and new drug development.
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Cardiotonic Agents
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chemistry
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pharmacology
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Energy Metabolism
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drug effects
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Heart
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drug effects
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physiopathology
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Humans
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Myocardial Contraction
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drug effects
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Panax
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chemistry
5.Clinical observation of axial offset after treatment by Ilizarov bone transport technology.
Jing-shuang WANG ; Si-bin HU ; Hong-hui SUN ; Ji-hui ZHENG ; Jun-fu ZHAO ; Dao-kuo LIU ; Liang LIN ; Hai-fenz DENG ; Yong-bo ZHANG
China Journal of Orthopaedics and Traumatology 2016;29(1):73-76
OBJECTIVETo observe the incidence, causes and deviation angle of axial offset in patients with fracture ununited treated by Ilizarov bone transport technology.
METHODSFrom January 2007 to December 2012, 10 patients with fracture ununited were treated by Ilizarov bone transport including 8 males and 2 females with an average age of (30.3 ± 10.6) years old ranging from 18 to 49 years old. The segment of bone defect involved upper tibial in 2 cases, medial tibia in 2 cases, lower tibial in 5 cases, upper femoral in 1 case. For Paley type of bone defect, 6 cases were type B1, 4 cases were B3. The incidence and deviation angle of axial offset after Ilizarov bone transport technology were observed and evaluated on bone result by Paley assessment.
RESULTSAll patients were followed up from 19 to 32 months with an average of (22.0 ± 5.6) months. Three cases were natural healed at fracture ends, the other 7 cases were healed after bone graft. The time of external fixator was 16 to 28 months. At the last follow-up, there were 3 cases occurred coronal angulation of angle 5° to 11° with an average of (8.7 ± 3.2). Sagittal angulation was in 4 cases, angle 6° to 9° with an average of (8.5 ± 2.1)°. There were 4 cases occurred axial offset. In the last follow-up, according to Paley evaluation criteria, osseous results were excellent in 7 cases, good in 3 cases; functional results were excellent in 6 cases, good in 4 cases.
CONCLUSIONAxial deviation after the Ilizarov bone transport treatment is relatively common, which will result in delayed healing of bone and poor limb alignment. In order to improve the bone healing, corresponding measurements should be taken to avoid or reduce the incidence of axial deviation during and after the operation.
Adolescent ; Adult ; Female ; Fracture Healing ; Fractures, Ununited ; surgery ; Humans ; Ilizarov Technique ; adverse effects ; Male ; Middle Aged
6.Competitive capacity of HIV-1 strains carrying M184I or Y181I drug-resistant mutations.
Jue LI ; Lin LI ; Han-ping LI ; Dao-min ZHUANG ; Si-yang LIU ; Yong-jian LIU ; Zuo-yi BAO ; Zheng WANG ; Jing-yun LI
Chinese Medical Journal 2009;122(9):1081-1086
BACKGROUNDVirus with nucleoside reverse transcriptase inhibitors (NRTIs) or nonnucleoside reverse transcriptase inhibitors (NNRTIs) resistant mutations show different evolution tendencies when the anti-viral therapies are interrupted. Understanding the replication fitness of drug-resistant virus is important for the study of the prevalence of drug-resistance. For this purpose, we characterized the replication capacity of HIV-1 virus carrying lamivudine (3TC) or nevirapine (NVP) resistant mutations.
METHODS3TC and NVP resistant variants were induced in vitro by selecting wild type virus in the presence of drugs. For the competitive replication assay, drug-resistant variants were cocultured with wild-type virus in the presence or absence of drugs. The ratios of the viral species were determined over time by using a real-time RT-PCR-based assay.
RESULTS3TC-resistant (M184I mutation) and NVP-resistant (Y181I mutation) virus should be selected in vitro in two different ways. The competitive replication assay showed that the ratio of virus carrying a M184I mutation increased from 98.8%, while the wild type virus decreased to 1.2% after 4 passages in the presence of 3TC; the percentage of virus carrying the Y181I mutation increased to 90.5%, while wild type virus decreased to 9.5% in the presence of NVP. In the absence of drugs, the ratio of virus carrying the M184I mutation decreased to 5.3%, while wild type virus increased to 94.7%; the ratio of virus carrying Y181I increased to 75%, while wild type virus decreased to 25% after 4 passages.
CONCLUSIONSThe NVP-resistant virus is fitter than wild type virus even in the absence of NVP that may be the reason that NNRTIs-resistant virus is spreading quickly.
Cell Line ; Drug Resistance, Viral ; genetics ; HIV-1 ; drug effects ; genetics ; growth & development ; physiology ; Humans ; Lamivudine ; pharmacology ; Mutation ; Nevirapine ; pharmacology ; Reverse Transcriptase Inhibitors ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Virus Replication ; genetics ; physiology
7.Allele-specific real-time PCR testing for minor HIV-1 drug resistance mutations: assay preparation and application to reveal dynamic of mutations in vivo.
Dong-Xing GUO ; Han-Ping LI ; Lin LI ; Dao-Min ZHUANG ; Li-Yan JIAO ; Zheng WANG ; Zuo-Yi BAO ; Si-Yang LIU ; Yong-Jian LIU ; Jing-Yun LI
Chinese Medical Journal 2010;123(23):3389-3395
BACKGROUNDIt is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20% - 30%. The aim of this study was to evaluate allele-specific real-time PCR (ASPCR) to detect the resistance-related mutations located at positions 103, 184 and 215.
METHODSWe developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140 clinical samples using this method.
RESULTSThe sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M184I, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.
CONCLUSIONSDrug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Y emerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.
Alleles ; Drug Resistance, Viral ; HIV-1 ; drug effects ; genetics ; Humans ; Mutation ; Real-Time Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity
8.Shenfu Injection suppresses inflammation by targeting haptoglobin and pentraxin 3 in rats with chronic ischemic heart failure.
Si-Dao ZHENG ; Hong-Jin WU ; Shao-Ping YU ; Jian-Xun REN ; Wei-Wei DUO ; Zeng-Chun MA ; Yue GAO ; Sheng-Qi WANG ; Yu-Na LIU
Chinese journal of integrative medicine 2015;21(1):22-28
OBJECTIVETo investigate the regulatory effects of Shenfu Injection (SFI, ) on hemodynamic parameters and serum proteins in rats with post-infarction chronic heart failure (CHF).
METHODSForty-five healthy Wistar rats were randomized into three groups: sham, heart failure (model) and SFI group. The CHF was induced by left coronary artery ligation. Seven days after the surgical operation, animals in the sham group and the model group received saline (6.2 mL/kg/d), while animals in the SFI group received SFI (6.2 mL/kg d) intraperitoneally. Four weeks later, cardiac hemodynamic parameters were measured via the carotid route. The expression of serum proteins was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS).
RESULTSRecording of hemodynamic parameters showed that left ventricular systolic pressure (LVSP), maximum rate of left ventricular pressure (+dp/dtmax) rise, and maximum rate of left ventricular pressure (-dp/dtmax) decrease, while the left ventricular end diastolic pressure (LVEDP) rose in the model group compared to those in the sham group (P <0.05). The results of the MALDI-TOF MS indicated that haptoglobin (HP), pentraxin 3 (PTX3) and alpha-1-antitrypsin were up-regulated, while serum albumin and 40S ribosomal protein were down-regulated in the model group (P <0.05). Compared with the model group, LVSP, +dp/dtmax and -dp/dtmax were higher, while LVEDP was lower in the SFI group (P<0.05). Expression levels of HP and PTX3 were lower than in the model group (P<0.05).
CONCLUSIONSFI could improve hemodynamic function and decrease inflammatory reactions in the pathophysiology of CHF. The serum proteins HP and PTX3 could be potential biomarkers for chronic ischemic heart failure, and they could also be the serum protein targets of SFI.
Animals ; Blood Proteins ; metabolism ; C-Reactive Protein ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Electrophoresis, Gel, Two-Dimensional ; Haptoglobins ; metabolism ; Heart Failure ; blood ; complications ; drug therapy ; physiopathology ; Heart Function Tests ; Hemodynamics ; Hydrogen-Ion Concentration ; Imaging, Three-Dimensional ; Inflammation ; complications ; drug therapy ; Male ; Myocardial Ischemia ; blood ; complications ; drug therapy ; physiopathology ; Phytotherapy ; Proteome ; metabolism ; Rats, Wistar ; Serum Amyloid P-Component ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
9.Regulatory Mechanisms of the Molecular Pathways in Fibrosis Induced by MicroRNAs.
Cui YANG ; Si-Dao ZHENG ; Hong-Jin WU ; Shao-Jun CHEN
Chinese Medical Journal 2016;129(19):2365-2372
OBJECTIVEMicroRNAs (miRNAs or miRs) play critical roles in the fibrotic process in different organs. We summarized the latest research progress on the roles and mechanisms of miRNAs in the regulation of the molecular signaling pathways involved in fibrosis.
DATA SOURCESPapers published in English from January 2010 to August 2015 were selected from the PubMed and Web of Science databases using the search terms "microRNA", "miR", "transforming growth factor β", "tgf β", "mitogen-activated protein kinase", "mapk", "integrin", "p38", "c-Jun NH2-terminal kinase", "jnk", "extracellular signal-regulated kinase", "erk", and "fibrosis".
STUDY SELECTIONArticles were obtained and reviewed to analyze the regulatory effects of miRNAs on molecular signaling pathways involved in the fibrosis.
RESULTSRecent evidence has shown that miRNAs are involved in regulating fibrosis by targeting different substrates in the molecular processes that drive fibrosis, such as immune cell sensitization, effector cell activation, and extracellular matrix remodeling. Moreover, several important molecular signaling pathways involve in fibrosis, such as the transforming growth factor-beta (TGF-β) pathway, mitogen-activated protein kinase (MAPK) pathways, and the integrin pathway are regulated by miRNAs. Third, regulation of the fibrotic pathways induced by miRNAs is found in many other tissues in addition to the heart, lung, liver, and kidney. Interestingly, the actions of many drugs on the human body are also induced by miRNAs. It is encouraging that the fibrotic process can be blocked or reversed by targeting specific miRNAs and their signaling pathways, thereby protecting the structures and functions of different organs.
CONCLUSIONSmiRNAs not only regulate molecular signaling pathways in fibrosis but also serve as potential targets of novel therapeutic interventions for fibrosing diseases.
Animals ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Fibrosis ; genetics ; metabolism ; Humans ; MicroRNAs ; genetics ; Mitogen-Activated Protein Kinases ; genetics ; metabolism ; Transforming Growth Factor beta ; genetics ; metabolism
10.Immunogenicities of Env glycoproteins from circulating HIV-1 isolates in China focusing on the strategy of "DNA prime plus protein boost".
Zheng WANG ; Shi-xia WANG ; Si-yang LIU ; Zuo-yi BAO ; Dao-min ZHUANG ; Lin LI ; Chun-hua ZHANG ; Lu ZHANG ; Jing-yun LI ; Shan LU
Chinese Medical Journal 2009;122(19):2339-2345
BACKGROUNDThe adenovirus-based HIV-1 vaccine developed by Merck Company suffered from an unexpected failure in September 2007. This generated a big shift in the strategy of HIV vaccine development with renewed focus on the induction of neutralizing antibodies. A major challenge in developing an HIV-1 vaccine is to identify immunogens and adopt delivery methods that can elicit broadly neutralizing antibodies against primary isolates of different genetic subtypes.
METHODSMost circulating HIV-1 isolates in China are composed of clades Thai-B, CRF_BC and CRF01_AE. In order to construct DNA vaccines against these 3 HIV-1 subtypes, DNA vaccines carrying the gp120 regions from HIV-1 isolates of GX48(AE), GX79(AE), NX22(BC), GS22(BC), HN24(Thai-B) were constructed. Expression of gp120 from these DNA vaccines was detected by Western blotting in transiently transfected 293T cells. Pilot immunizations of New Zealand white rabbits were performed using the strategy of "DNA prime plus protein boost" and the neutralizing antibody response was detected in a Tzm-bl cell based assay against different HIV-1 strains.
RESULTSResponse of gp120-specific antibody was relatively low after DNA primes (mean titer = 10(4.72)); however, the titer of gp120-specific antibody went up with 2 protein boosts (mean titer = 10(6.81)). Above all, neutralizing antibody (Nab) titers induced by this combined approach were much better than those elicited by DNA or protein used alone (P < 0.01). Neutralizing activities of immunized rabbit sera against several pseudoviruses and laboratorial strains were evaluated, most rabbit sera primed with monovalent vaccine were capable of neutralizing only 1 of 5 viruses, however, sera primed with the polyvalent DNA vaccines were able to neutralize at least 2 of 5 viruses.
CONCLUSIONPolyvalent DNA prime plus protein boost is an effective immunization strategy to broaden the neutralization breadth and further research should be performed on the basis of this pilot study.
AIDS Vaccines ; immunology ; Animals ; Antibodies, Neutralizing ; blood ; Female ; HIV Envelope Protein gp120 ; genetics ; immunology ; Humans ; Immunization ; Immunoglobulin G ; blood ; Phylogeny ; Rabbits ; Vaccines, DNA ; immunology