IL-31 expression in HIV-infected patients with different routes of disease transmission.

ABSTRACT: Acquired immunodeficiency syndrome (AIDS) is caused by the human immunodeficiency virus (HIV). AIDS is characterized by an impaired immune system and low cellular immunity. The main manifestation of AIDS is a reduction in the number of CD4+ T cells and alteration in cytokine concentration. The present work aimed to explore the expression of IL-31 in HIV infection and disease progression.Serum samples were collected from HIV-infected patients with different routes of disease transmission. The subjects included 24 patients who were infected with HIV upon blood transmission and 36 patients who had acquired the disease through sexual transmission (21 cases of homosexual transmission and 15 cases of heterosexual transmission). In addition, 20 normal healthy individuals were included to serve as the control group. The levels of IL-31 in the collected serum samples were estimated using the human IL-31 Platinum ELISA kit.The serum analysis results revealed that the concentration of IL-31 in the serum samples for the blood transmission, sexually transmission, and normal group patients was 4.07 ±â€Š1.63 pg/L, 7.43 ±â€Š1.15 pg/L, and 2.87 ±â€Š1.04 pg/L, respectively. The statistical analysis revealed that the concentration of IL-31 in HIV-1 infection was higher than that in the normal control. In addition, the expression of IL-31 was significantly higher in the sexual transmission group compared to the blood transmission group (P < .05).IL-31 could have an important role in HIV infection, although the role of IL-31 in disease progression in HIV-infected individuals requires further research.

M6A-Mediated Upregulation of LINC00106 Promotes Stemness and Metastasis Properties of Hepatocellular Carcinoma via Sponging Let7f.

Background: Hepatocellular carcinoma (HCC) cells exhibit the stemness property, which makes the patient with HCC prone to tumor recurrence and metastasis. Despite the prominent regulatory role of long non-coding RNAs (lncRNAs) in tumor stemness, the roles and molecular mechanisms of LINC00106 in HCC are poorly understood. Methods: LINC00106, let7f and periostin expression levels in tissue specimens and cell lines were assessed through qRT-PCR and immunohistochemistry (IHC). Various in vivo and in vitro assays, namely sphere/colony formation, proportion of side population cells (SP%), invasion, migration, western blot, and murine xenograft model were employed for assessing the stemness and metastatic properties of HCC cells. Luciferase reporter assays, RNA-seq, RNA pull-down, RNA immunoprecipitation (RIP) were conducted to clarificate the target gene and analyze the underlying mechanisms. Results: LINC00106 was prominently upregulated in tissues and cell lines of HCC. Patients having a high LINC00106 level exhibited a poor outcome. Under in vivo and in vitro conditions, the stemness and metastatic properties of HCC cells were augmented by LINC00106. Additionally, LINC00106 was found to sponge let7f to upregulate periostin, which lead to the activation of periostin-associated PI3K-AKT signaling pathway. Moreover, m6A methylation was found to cause LINC00106 upregulation while maintaining LINC00106 RNA transcript stability. Conclusion: m6A methylation triggers the upregulation of LINC00106, which promotes the stemness and metastasis properties in HCC cells by sponging let7f, thereby resulting in periostin activation. The findings indicate the potential of LINC00106 as a diagnostic marker and therapeutic target for HCC.

Linc8986 and linc0597 in plasma are novel biomarkers for systemic lupus erythematosus.

Despite increasing evidence that large intergenic non-coding RNAs (lincRNAs) are widely involved in human disease, the role of lincRNAs in the development of systemic lupus erythematosus (SLE) has remained largely elusive. The purpose of the present study was to investigate the expression of three lincRNAs (linc0597, linc8986 and linc7190) in the plasma of patients with SLE and their potential use as biomarkers for the diagnosis and treatment of SLE. Plasma samples were obtained from 54 patients with SLE, 24 patients with rheumatoid arthritis (RA), 24 patients with Sjogren's syndrome (SS) and 22 healthy controls. LincRNA expression levels were measured by reverse transcription-quantitative PCR. Compared with those in the healthy controls, the plasma levels of linc0597 and linc8986 were significantly increased in the patients with SLE (P<0.001), while the difference in the level of linc7190 was not significant (P=0.052). In addition, there was no significant difference in the levels of linc0597 and linc8986 among patients with RA, patients with SS and the healthy controls (P>0.05). Compared with patients with SLE without lupus nephritis (LN), the levels of linc0597 were significantly higher in patients with LN (P=0.044). For linc7190 and linc8986, there was no significant difference between patients with and without LN (P>0.05). Furthermore, complement component 3 (C3) levels were used to evaluate whether the expression of linc8986 and linc0597 is related to the activity of SLE. The results indicated that the levels of linc8986 and linc0597 were negatively correlated with the level of C3 (P<0.001 and P=0.004, respectively). Further analysis suggested that linc0597 and linc8986 were able to specifically identify patients with SLE and that a combination of linc0597 and linc8986 may improve the diagnostic accuracy. Therefore, the plasma levels of linc0597 and linc8986 may be suitable biomarkers for diagnosing SLE.

The genotype analysis of the hepatitis C virus in Heilongjiang Province, China.

ABSTRACT: Introduction: Hepatitis C virus (HCV) infection is a major public health issue. HCV genotype identification is clinically important to tailor the dosage and duration of treatment, and recombination in intra-patient populations of HCV may lead to the generation of escape mutants, as previously observed for other RNA viruses. Up to now, there is no study assessing HCV genotypes and subtypes in Heilongjiang Province, China.Methods: To determine genotype and phylogenetic analysis of HCV in Heilongjiang Province is crucial. In this study, we amplified 3 genome regions (5'UTR, E1, and NS5B) of 30 HCV patients in Heilongjiang Province, amplified products were analyzed by bioinformatics.Results: We found that 23 specimens had concordant subtypes in the 3 gene regions (2a and 1b), 7 HCV patients were considered the recombinants, the recombination pattern of the 7 HCV patients in the 5'UTR, E1, and NS5B region as followed: 1b/2a/1b, 2a/2a/1b, 1b/2a/2a, 1b/2a/1b, 1b/2a/1b, 1b/2a/1b, 2a/2a/1b.Conclusions: The findings in the present study showed that a higher recombination rate (23%) than other researches, and the recombination of 2a/1b in the 5'UTR, E1, and NS5B region was only found in the present study up to now.

Anti-human serum albumin autoantibody may be involved in the pathogenesis of autoimmune bullous skin diseases.

Although effective immunological diagnostic systems for autoimmune bullous skin diseases (AIBD) have been established, there are still unidentified cutaneous autoantigens. The purpose of this study is to investigative whether anti-human serum albumin (HSA) autoantibodies exist in AIBD sera and their potential pathogenesis. By immunoprecipitation-immunoblotting, immunofluorescence assay, anti-HSA autoantibodies could be detected in AIBD sera; by ELISAs, positive rates of AIBD sera for IgG and IgA anti-HSA autoantibodies were 29% and 34%, respectively. The IgG anti-HSA autoantibodies in ABID sera recognized a number of HSA antigen epitopes and therefore a polyclonal antibody against HSA were next employed to study its pathogenesis. In vitro cell and tissue culture models, anti-HSA antibody could influence DNA damage-related signaling proteins, via activation of phospho-p38 signaling pathway. This is the first report that an autoantibody may influence DNA damage-related signaling proteins. Statistical analyses also proved that anti-HSA autoantibodies were positively correlated with various known autoantibodies and clinical features of ABID patients. In summary, IgG and IgA autoantibodies to HSA may have diagnosis values for AIBD. DNA damage-related signaling proteins might be involved in the pathogenic role of anti-HSA autoantibodies in AIBD. Phospho-p38 signaling pathway is a potential target for treatment of AIBD positive for serum anti-HSA autoantibodies.

Changes of intestinal flora in patients with systemic lupus erythematosus in northeast China.

OBJECTIVE: The human gut harbors diverse microbes that play a fundamental role in the well-being of their hosts. Microbes can cause autoimmunity, trigger autoimmunity in genetically susceptible individuals or prevent autoimmunity. There were reports about intestinal flora changes in Systemic Lupus Erythematosus (SLE) patients, but no data were available in northeast China. In this study, we investigated the intestinal flora changes of SLE patients in Heilongjiang province located in northeast China. METHODS: Feces from 16 SLE patients and 14 healthy volunteers were employed to extract bacterial DNA, amplify 16s RNA of bacteria, and analyze the biological information by sequencing. The statistical analysis used the SPSS version of 17. RESULT: We found that there were 1 phylums, 4 families and 9 genera in the intestinal flora of SLE patients. And the nine differences genera can be used to distinguish SLE patients from normal people. CONCLUSION: We found an increase of Proteobacteria and a decrease of Ruminococcaceae in SLE patients in different regions. In addition, we found that some proteins, enzymes, and diseases were significantly associated with SLE.

Serum vitamin D levels among children aged 0-12 years in the First Affiliated Hospital of Harbin Medical University, China.

Background: Vitamin D deficiency (VDD) and vitamin D insufficiency (VDI) are highly prevalent in the world, but the vitamin D status of children in northeast China is seldom investigated. The aim of this study was to clarify the prevalence of VDD and VDI among children in the First Affiliated Hospital of Harbin Medical University in Heilongjiang province in China. Methods: We collected data from 9795 children who were outpatients aged 0-12 years who visited the First Affiliated Hospital of Harbin Medical University from September 2014 to August 2016. Serum 25-hydroxy vitamin D [25(OH)D] levels were determined by chemiluminescent immunoassay and categorized as <20, 20-30 and >30 ng/mL. Results: The highest mean level of serum 25(OH)D was found at the 1-3 years stage (31.14 ng/mL) and the lowest at 6-12 years stage (18.58 ng/mL). The mean serum 25(OH)D level among school girls (17.86 ng/mL) was lower than that of boys (19.12 ng/mL). The prevalence of vitamin D sufficiency during 2014 was only 17.2%, but increased to ~45% in 2016. Conclusions: The prevalence of VDD and insufficiency among children in the First Affiliated Hospital of Harbin Medical University is high, especially among children aged 6-12 years.

Laminins in an in vitro anterior lens capsule model established using HLE B-3 cells.

Cataracts are the most common eye disease to cause blindness in patients. The abnormal deposition of laminins (LMs) in the lens capsule and the disruption of capsular epithelium contribute to cataract development, although the mechanism by which this occurs is currently unclear. The present study aimed to reproduce HLE B­3 basement membranes (BMs) using HLE B­3 cells and to analyze the similarities of LM expression between HLE B­3 BMs and human anterior lens capsule (ALC). Immunohistochemistry (IHC), ELISA, western blot analysis and immunoprecipitation (IP)­western blot analysis were used to detect total LMs, LM trimers and 11 LM subunits in HLE B­3 cells, HLE B­3 BMs and human ALCs. In IHC staining, HLE B­3 cells and human ALCs were positive for LMs. In LM ELISA, all samples analyzed were positive for LMs. Western blot analysis detected all LM subunits except for LMγ3 in HLE B­3 cell lysate, 4 subunits (LMα4, LMα2, LMα1 and LMγ1) in HLE B­3 cell culture supernatant, 5 subunits (LMα4, LMα2, LMα1, LMß3 and LMγ1) in HLE B­3 BMs, and 3 subunits (LMα4, LMγ2 and LMγ1) in human ALCs. The results of IP­western blot analysis revealed that the LM411 trimer was detected in HLE B­3 cell culture supernatant. These results indicated that HLE B­3 BMs were similar to human ALCs in terms of LM expression. Therefore, HLE B­3 BMs could be used as an in vitro ALC model to determine the role of LMs in ALC in the pathogenesis of cataracts and to select potential anti­cataract drugs.

Position 22 of the V3 loop is associated with HIV infectivity.

Human immunodeficiency virus subtype 1B (HIV-1B) binds to the CD4 receptor and co-receptor CCR5 or CXCR4 to enter T lymphocytes. The amino acid sequence of the HIV envelope glycoprotein V3 region determines the co-receptor tropism, thereby influencing the infectivity of the virus. Our research group previously found that the amino acid at position 22 of the V3 region may affect the infectivity of the virus, and in this study, we tested this hypothesis. We constructed pseudoviruses by changing the amino acids at position 22 of the V3 region in CCR5-tropic and CXCR4-tropic viruses and tested their infectivity. When the amino acid at V3 position 22 was altered in the CCR5- and CXCR4-tropic viruses, their ability to infect cells decreased to 20.6% and 17.14%, respectively. Therefore, we propose that residue 22 in the V3 region of subtype HIV-1B significantly influences the infectivity of the virus.

Variation analysis of E1 and E2 in HCV subtypes.

Pegylated interferon and ribavirin combination therapy effectively suppresses viral replication in 50 %-60 % of hepatitis C virus (HCV)-infected patients. However, HCV-infected patients often display varied responses to therapy, and strains of subtype lb (the most widespread HCV subtype worldwide) have more-severe clinical manifestations, greater viral loads, and poorer responses to interferon treatment. Therefore, understanding the genomic variability of HCV is crucial to treatment of HCV infection. In this study, we used the appropriate software to analyze the nucleotide, and amino acid sequences of the envelope proteins (E1 and E2) of HCV to investigate the extent of their variability in several HCV subtypes (1a, 1b, 2a, 2b, 3a, 4a, 5a and 6a) and calculated the ratio of nonsynonymous to synonymous substitutions (dN/dS) in these proteins to investigate the immunological pressure acting on them. We also predicted the N-glycosylation sites in E1 and E2 to determine their association with viral neutralization. We found that E1 is more variable, has a higher dN/dS ratio, and has more N-glycosylation sites than E2 in HCV subtype 1b. This indicates that the variability of E1, its dN/dS ratio, and its degree of N-glycosylation might play an important role in the treatment of infection with HCV subtype 1b.

Trends in HIV infection in the First Affiliated Hospital of Harbin, China.

BACKGROUND: Major hospitals in most Chinese cities have the capability to perform HIV testing. However, it is not a routine test for all patients and, as a result, many patients are not aware of their HIV status. To understand the rate of HIV infection and the factors associated with infection, we tested serum to determine HIV status and analyzed factors associated with HIV infection. METHODS: We collected blood samples from 348,151 patients who visited the First Affiliated Hospital of Harbin Medical University from 1 January 2007 to 31 December 2012. Serum was screened with an ELISA. If the test was positive, we conducted two additional ELISAs: a repeat with the initial kit and one with another kit. If there was a positive result with either of these two ELISA kits, western blotting was performed at Harbin Centers for Disease Control and Prevention. RESULTS: The HIV positivity rate of inpatients significantly increased during the course of this study. HIV infection in patients appeared to differ by sex, age, occupation, marital status, educational level, and route of infection. HIV was more prevalent in men than in women. More than 80% of HIV-positive patients had not received higher (>12 years) education. From 2007 to 2012, HIV-positive patients were mainly infected through sexual transmission. For sexually acquired infections, the rate of HIV infections through homosexual contact has increased rapidly in recent years, and ranged from 36.4% to 65.1%. CONCLUSIONS: The number of patients diagnosed as HIV positive has increased in recent years. Offering routine HIV testing in hospitals is feasible and can increase linkage to HIV care and treatment for many individuals.

Toll-like receptor 7 promotes the apoptosis of THP-1-derived macrophages through the CHOP-dependent pathway.

Macrophage apoptosis is a prominent characteristic of advanced atherosclerotic plaques and leads to plaque destabilization. Certain studies have confirmed that influenza virus A (IVA) infection is related to acute myocardial infarction (AMI). However, it remains unknown as to whether this phenomenon is associated with Toll-like receptor (TLR)7, since single-stranded RNA (ssRNA) of IVA is a natural ligand of TLR7. Thus, in the present study, THP-1­derived macrophages were infected with IVA or treated with imiquimod (IMQ) in the presence or absence of pre-treatment with oxidized low-density lipoprotein (oxLDL). The macrophages were pre-treated with oxLDL (5 µg/ml) for 24 h to mimic high lipid conditions. Cell viability and apoptosis were detected by 3-(4,5-dimethylthiazol-2-y-1)­2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and flow cytometry, respectively. Our results revealed that TLR7 played an important role in macrophage apoptosis and cytokine secretion. Both IVA infection and IMQ treatment increased TLR7 expression, as well as the secretion of pro-inflammatory cytokines [interleukin (IL)-6, monocyte chemotactic protein (MCP)-1] and apoptosis. However, this increase in cytokine secretion occurred independently of cell apoptosis. oxLDL had potential synergistic pro-apoptotic effects combined with TLR7 activation. To determine whether endoplasmic reticulum (ER) stress plays a role in cell apoptosis, the mRNA and protein expression of known markers of ER stress [glucose-regulated protein (GRP)78 and C/EBP homologous protein (CHOP)] was detected by reverse transcription PCR (RT-PCR), quantitative reverse transcription PCR (qRT-PCR) and western blot analysis. Our results revealed that apoptosis aggravated ER stress, as shown by the overexpression of the pro-apoptotic sensor, CHOP. In conclusion, our study demonstrates the converging role of oxLDL pre-treatment, IVA infection and IMQ in ER stress-induced cell apoptosis.

Self-medication of psoriasis in southwestern China.

BACKGROUND: Medication adherence is essential for psoriasis management. Although studies have determined the prevalence and correlative factors of non-adherence, as far as we know, there has been no study specifically addressing them in China. METHODS: Anonymous 23-item questionnaires were distributed to 324 psoriatic patients from June 2012 to June 2013. χ(2) test, normality test, Student's t test or Mann-Whitney U test, Bonferroni correction and the binary logistic regression model were applied. RESULTS: Self-medication of psoriasis was high in China (82.4%). Joint involvement, absence of communication with families and short communication duration with doctors were the three top factors affecting adherence. Besides low efficacy (16.0%) and recurrence frustration (27.0%), we found that patients' deception by sham advertisings (24.1%) is an non-negligible issue. CONCLUSION: Psoriasis self-medication in China was high. Improvement of patients' communication with families and/or doctors were suggested to be crucial to enhance adherence. Meanwhile, extermination of sham advertisings by administrative agencies is of great concern.

Frequency of primary glomerular disease in northeastern China

Most frequently reported Chinese renal biopsy data have originated from southeastern China. The present study analyzed the renal biopsy data from northeastern China. The records of 1550 consecutive native patients who were diagnosed with primary glomerular diseases (PGD) after renal biopsy at our hospital during 2005-2009 were used. These patients were divided into four age groups for stratified analysis: <15, 15-44, 45-59, and ≥60 years old. Among PGD, minimal change disease (MCD) was the most common histologically diagnosed disease (30.7 percent), followed by IgA nephropathy (IgAN), mesangial proliferative glomerulonephritis (MsPGN), membranous nephropathy (MN), membranoproliferative glomerulonephritis (MPGN), focal segmental glomerulosclerosis (FSGS), and endocapillary proliferative glomerulonephritis (EnPGN). MCD was the disease most frequently observed (43.7 percent) in the <15-year-old group. MsPGN was the most common disease in the elderly group (38.1 percent). MsPGN was more prevalent in females (27.8 percent), whereas MCD was more prevalent in males (35.3 percent). Primary glomerular diseases constituted the most commonly encountered group of diseases with a high prevalence of MCD, which predominantly affected males and young adults. The prevalence of MCD was high in northeastern China. Further study is necessary to expand the epidemiologic data available for renal disease in China.

Frequency of primary glomerular disease in northeastern China.

Most frequently reported Chinese renal biopsy data have originated from southeastern China. The present study analyzed the renal biopsy data from northeastern China. The records of 1550 consecutive native patients who were diagnosed with primary glomerular diseases (PGD) after renal biopsy at our hospital during 2005-2009 were used. These patients were divided into four age groups for stratified analysis: <15, 15-44, 45-59, and ≥60 years old. Among PGD, minimal change disease (MCD) was the most common histologically diagnosed disease (30.7%), followed by IgA nephropathy (IgAN), mesangial proliferative glomerulonephritis (MsPGN), membranous nephropathy (MN), membranoproliferative glomerulonephritis (MPGN), focal segmental glomerulosclerosis (FSGS), and endocapillary proliferative glomerulonephritis (EnPGN). MCD was the disease most frequently observed (43.7%) in the <15-year-old group. MsPGN was the most common disease in the elderly group (38.1%). MsPGN was more prevalent in females (27.8%), whereas MCD was more prevalent in males (35.3%). Primary glomerular diseases constituted the most commonly encountered group of diseases with a high prevalence of MCD, which predominantly affected males and young adults. The prevalence of MCD was high in northeastern China. Further study is necessary to expand the epidemiologic data available for renal disease in China.

Improved expression of secretory and trimeric proteins in mammalian cells via the introduction of a new trimer motif and a mutant of the tPA signal sequence.

Ideal immunogenicity in antigens is a prerequisite to eliciting a sufficiently strong immune and memory response via either DNA or protein vaccines. To improve immunogenicity, efforts have focused on high-level expression of target proteins and on maintaining their natural conformations. In the present work, two trimer motifs (MTQ and MTI) were designed and introduced into a plasmid vector with the tissue plasminogen activator signal peptide (tPA-SP). Next, we examined the efficacy and the efficiency of the two motifs as well as the introduction of tPA-SP and its mutant forms, 22P/A and 22P/G, in facilitating the secretory expression of trimeric proteins in mammalian cells. We found that both trimer motifs could produce the target protein in a trimeric form at a high level. Introduction of tPA-SP 22P/A markedly increased the secretory expression level. The combination of the trimer motif, MTQ, and the signal peptide, 22P/A, may serve as a universal mammalian vector for producing trimeric proteins in vaccine development.

Position 22 of the V3 loop is associated with co-receptor usage and disease progression in HIV-1 subtype B isolates.

Our goal in this study was to analyze position 22 of the V3 loop associated with co-receptor usage and disease progression in human immunodeficiency virus type 1 (HIV-1) subtype B infection. Bioinformatics approaches were used to compare the amino acid sequence and secondary structure of the V3 loop of the CCR5-tropic virus and CXCR4-tropic virus in HIV-1 subtype B. HIV-1 subtype B V3 amino acid sequence files in the FASTA format were collected from the HIV Sequence Database. The amino acid sequences of different tropism were multiple-aligned with CLUSTAL W program, and the frequencies of the amino acids at each position of the V3 loop sequences of two groups were calculated and sorted in descending order. The secondary structure of the consensus V3 amino acid sequences from CCR5-tropic and CXCR4-tropic viruses were predicted with the APSSP2 method. The amino acids at positions 11, 22, and 25 of V3 were different between the CCR5-tropic virus and CXCR4-tropic virus. The consensus amino acid frequencies were found to be 71.9% S, 66.7% A, and 56.0% D for the CCR5-tropic virus and 50.0% R, 57.1% T, and 26.2% Q for the CXCR4- tropic virus at positions 11, 22, and 25, respectively. There was a strong association between the identity of the residues at position 11, 22, and 25 of the V3 loop amino acid sequence and CD4+ T cell counts of different patients. The change of the residue at position 22 in the R5-tropic or X4-tropic viruses is expected to likely change the secondary structure to be similar to the X4-tropic or R5-tropic viruses. Our study indicates that position 22 of the V3 loop amino acid sequence is significantly associated with viral tropism and disease progression in HIV-1 subtype B.

Biosorption of cadmium(II), zinc(II) and lead(II) by Penicillium simplicissimum: Isotherms, kinetics and thermodynamics.

The isotherms, kinetics and thermodynamics of Cd(II), Zn(II) and Pb(II) biosorption by Penicillium simplicissimum were investigated in a batch system. The effects of pH, initial metal ions concentration, biomass dose, contact time, temperature and co-ions on the biosorption were studied. Adsorption data were well described by both the Redlich-Peterson and Langmuir model. Chemical ion-exchange was found to be an important process based on free energy value from Dubini-Radushkevich isotherm for all metal ions. The results of the kinetic studies of all metal ions at different temperature showed that the rate of adsorption followed the pseudo second-order kinetics well. The thermodynamics constants DeltaG degrees , DeltaH degrees and DeltaS degrees of the adsorption process showed that biosorption of Cd(II), Zn(II) and Pb(II) ions on Penicillium simplicissimum were endothermic and spontaneous.

Subtype and sequence analysis of HIV-1 strains in Heilongjiang Province.

BACKGROUND: Human immunodeficiency virus (HIV-1) is divided into two types, HIV-1 (groups M, N and O) and HIV-2. Heilongjiang Province located in the northeast of China, and the feature of the subtype distribution and sequence characteristics of HIV-1 strains prevalent in Heilongjiang Province is still uncertain. The aim of this study was to investigate the subtype distribution and genetic characteristics of HIV-1 strains in one hospital in Heilongjiang Province. METHODS: HIV-1 env gene was amplified by nested-PCR from peripheral blood mononuclear cells (PBMCs) obtained from 19 HIV-1 seropositive individuals in Heilongjiang Province. The C2-V3 region was sequenced. Aligned the nucleotide sequence of 19 samples with CLUSTAL W (BioEdit) software, results were acquired and used for phylogenetic tree analysis after artificial adjustment. Reference sequence, downloaded from Los Alamos HIV Sequence Database, was used to identify the subtype of obtained sequence. Genetic distance between sequences was assessed using the software MEGA 3.1 Kimura 2-parameter, and the Phylogenetic tree was reestablished with Neighbor-Joining method. RESULTS: Phylogenetic tree analysis showed that 19 Heilongjiang strains clustered closely to subtype B strain from Thailand and were far from other international subtype reference strains. Statistical test showed no significant discrepancy between the genetic distance of interclass and intra-class (P > 0.05). The analysis of V3 loop amino sequence of 19 Heilongjiang B strains revealed that V3 tip motif of 10 samples (52.63%) was GPGQ, and of 4 samples (21.53%) was GPGR. CONCLUSIONS: The subtype of 19 HIV-1 seropositive individuals in Heilongjiang Province is B', and it is introduced from He'nan Province. V3 tip motifs of the HIV-1 isolates are mainly GPGQ and GPGR.