OBJECTIVETo establish NB4/VEGF-C cells xenograft in nude mice model, and explore the effect of VEGF-C on hematological malignancies

METHODSNB4/VEGF-C or NB4/pcDNA3.1 cell lines were established by transfecting the recombinant pcDNA3.1-VEGF-C plasmid and the vacant pcDNA3.1 vector into NB4 cells. The recombinant VEGF-C was identified by RT-PCR and Western blotting. Eighteen male BALB/c nude mice aged 4 - 5 weeks were equally divided into two groups. Mice irradiated by 4 Gy ⁶⁰Co were subcutaneously injected with 1 × 10⁷NB4/VEGF-C or NB4/pcDNA3.1 cells into one side of axilla. The volumes of xenograft tumor was evaluated according to L × t² × 0.52. Microvessel density (MVD) on the xenograft tumor section was detected by IHC with VWF antibody.

RESULTSNB4 cell xenograft tumors were developed in all mice of both the two groups. The growth of NB4/VEGF-C cells in nude mice was faster than in controls. There were statistically significant differences in the volume and weight of xenograft tumor between NB4/VEGF-C and NB4/pcDNA3.1 cell groups \[(631.44 ± 114.42) mm³ vs (491.22 ± 70.05) mm³\] (P = 0.006) and \[(321.78 ± 27.84) mg vs (288.57 ± 40.12) mg\] (P = 0.031), respectively. MVD in xenograft tumor of NB4/VEGF-C cells \[(50.8 ± 11.7)/mm²\] was higher than that in controls \[(18.9 ± 7.0)/mm²\] (P = 0.021). The Bcl-2 protein level in NB4/VEGF-C cells xenografts was higher than that in controls.

CONCLUSIONVEGF-C could promote proliferation of NB4 cells by inducing angiogenesis and inhibit cells apoptosis by upregulating antiapoptotic Bcl-2 protein expression in NB4 cells xenograft tumor.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Xenograft Model Antitumor Assays

BACKGROUNDGenetic factors are believed to play a role in the individual susceptibility to chronic obstructive pulmonary disease (COPD). A single nucleotide polymorphism (SNP) of tumour necrosis factor-alpha (TNF-alpha) has been reported but inconsistent results may arise from different populations and phenotypes of COPD. There are only a few published studies of interleukin-13 (IL-13) SNPs on COPD. The SNPs of TNF-alpha and IL-13 have not been studied in the Chinese population. This research was conducted to study the frequencies of IL-13 gene promoter 1055 (IL-13-1055) and TNF-alpha gene-308 polymorphisms in the patients with COPD and to investigate the effect of those genetic polymorphisms on COPD in the Chinese population.

METHODSA cohort of COPD patients and age matched controls were recruited from an inpatient hospital service in Beijing. Venous blood was obtained and genomic DNA was extracted from peripheral blood monocytes using standard method. Genomic DNA was used as a template for amplification by polymerase chain reaction (PCR) to determine the polymorphism at -1055 in the IL-13 gene promoter region. PCR restriction fragment length polymorphism (RFLP) was used to determine polymorphisms in the TNF-alpha gene-308 position. The products were investigated by sequence analysis also.

RESULTSOne hundred and eleven COPD patients and 97 controls were studied. Seventy-five cases were current smokers in COPD patients and 36 were current smokers in controls. The frequencies of TT genotype in the IL-13 gene promoter region were 11.7% (13/111) in the COPD group and 13.4% (13/97) in the controls (P = 0.713). However, the OR value of TT genotype was significantly increased to 6.4 (95% CI 1.62 - 25.39) in the smokers with COPD. TT genotype was also positively related to family history of COPD, OR = 7.7 (95% CI 1.37 - 43.80). The frequencies of A allele in the TNF-alpha gene were 5.9% in COPD and 3.1% in controls (P = 0.131). The OR value of A allele was 5.0 (95% CI 1.011 to 25.059) in smokers with COPD.

CONCLUSIONSThere is no significant difference in the frequencies of the TT genotype of IL-13-1055 or the A allele of the TNF-alpha between Han Chinese patients with COPD versus control. Thus, it does not appear that these SNPs are independent factors in COPD for Han nationality in Beijing. However, these SNPs may increase the risk of COPD among smokers.

Adult ; Aged ; China ; ethnology ; Female ; Genotype ; Humans ; Interleukin-13 ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Pulmonary Disease, Chronic Obstructive ; genetics ; immunology ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo explore the biological effect on NB4 cells proliferation, all-trans retinoic acid (ATRA) inducing differentiation and resistance to apoptosis by vascular endothelial growth factor (VEGF)-C cDNA transfection.

METHODSThe recombinant eukaryotic expression plasmid pcDNA3.1-VEGF-C and the vacant pcDNA3.1 vector were introduced separately into NB4 cells by lipofectamine mediation. The positive clones were screened by G418 and identified by reverse transcriptase-PCR (RT-PCR) and Western blotting. The proliferation capacity of NB4/VEGF-C cells was analysed by MTT assay and colony forming assay in vitro. After NB4/VEGF-C cells were induced by ATRA, the expression level of C/EBPalpha gene, CD11b on cells surface and morphological alteration were analysed by real-time quantitative PCR (RQ-PCR), flow cytometry (FCM), and Wright-Giemsa staining, respectively. FCM Annexin V-FITC/PI dual labeling technique was performed to investigate the etoposide (Vp16) induced NB4/VEGF-C cells apoptosis and bcl-2 gene expression level in these cells was analysed by RQ-PCR. The NB4/pcDNA3.1 cells was used as control in the above experiments.

RESULTSA stable NB4 cell line that secrets VEGF-C and its control lines were established. The proliferation capacity of the former was stronger than that of the latter. The expression level of C/EBPalpha gene of NB4/VEGF-C cells on ATRA treatment was only 1/32 that of NB4/pcDNA3.1 cells. The CD11b level and the degree of differentiation of NB4/VEGF-C were weaker than that of NB4/pcDNA3.1 cells. The percentage of apoptotic NB4/VEGF-C cells induced by Vp16 [(7.20 +/- 2.52)%] was significantly lower than that of NB4/pcDNA3.1 cells [(16.07 +/- 3.58)%] (P = 0.005), but the bcl-2 gene expression level of NB4/VEGF-C cells is 2.28-fold that of NB4/pcDNA3.1 cells.

CONCLUSIONThe VEGF-C via VEGFR-3 signaling pathway could promote the proliferation of leukemic cells by autocrine pathway and inhibit the cell differentiation mediated by ATRA and chemotherapy-induced apoptosis. VEGF-C/VEGFR-3 signaling loops might play an important role in disease progression and be potential therapeutic target for the treatment of leukemias.

Apoptosis ; drug effects ; genetics ; physiology ; Blotting, Western ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; metabolism ; CD11b Antigen ; genetics ; metabolism ; Cell Differentiation ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; genetics ; Drug Resistance, Neoplasm ; Flow Cytometry ; Genetic Vectors ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Tretinoin ; pharmacology ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; physiology

Objective To evaluate the development of the sphincter muscle complex(SMC)and defecation function in pediatric patients with congenital anorectal malformations(ARM).Methods A total of 64 children underwent MRI,among whom 39 were patients with ARM,and the others were patients without ARM undergoing MRI because of other dieases.The dimensions of the SMC in different planes were evaluated with different sequences and coils.The relationship between the SMC development and the defecation function was investigated.Results In control group,the absolute value of SMC width was (3.63?0.22)mm,which had a high correlation with age(r=0.998,P0.05).The SMCs in intermediate ARM patients[muscle index(MI)=0.47?0.05]and low ARM patients(MI=0.49? 0.05)were well developed.The SMCs in a portion of patients with high ARM(MI=0.28?0.06)were poorly developed,when MI≤0.18,anorectal contraction pressurewas significantly lower(t=3.55, P0.18[(0.85?0.20)vs(2.24?1.02)kPa].The length of anal canal with high-pressure[(10.88?3.64)vs(20.26?4.34)mm]was shorter(t=5.18,P0.18,the anorectal angle was less than 90 degrees,and normal continent function was found in 21 of 23 cases(91%).Conclusion MRI can be employed to evaluate the development of SMC in patients with ARM,MI was an objective criteria to evaluate the development of SMC.When MI≤0.18, maldevelopment of SMC will be highly suspected.

BACKGROUNDMen who have sex with men (MSM) have been impacted by HIV and now as an important driver of the HIV/AIDS epidemic in China. This study collected HIV sentinel surveillance system data on the MSM population to describe the characteristics and trends of the HIV epidemic among MSM in China from 2003 to 2011.

METHODSData on HIV prevalence and risk behaviors from 2003 to 2011 were obtained from the national HIV sentinel surveillance database.

RESULTSMSM sentinel surveillance data for 2011 showed that proportions of MSM who consistently used condoms during anal sex in the last six months and at last anal sex encounter were 43.3% and 74.1%, respectively. Between 2005 and 2011 there were no significant changes in the proportion of consistent condom use. The proportion of MSM who had multiple male sex partners in the last six months increased. Overall HIV prevalence over the years showed a rising trend from 0.9% in 2003 to 6.3% in 2011. The syphilis antibody positive rate was 7.8% in 2011. In addition, the proportion of MSM in heterosexual marriages was rising, AIDS awareness was also increasing, and the proportion of MSM who had taken an HIV test in the last year and was aware of the results rose. The proportion who had received intervention services in the last year stabilized.

CONCLUSIONSHIV prevalence in MSM populations is rapidly and widely distributed. MSM who are in heterosexual marriages may be a bridge group transmitting HIV to their heterosexual partners. The risky sexual behavior among MSM will remain a serious and important driver of China's AIDS epidemic over the next period of time. Intervention need to be strengthened, as does the effective implementation of measures to control AIDS and prevent it from spreading further.

Adolescent ; Adult ; China ; epidemiology ; HIV Infections ; diagnosis ; epidemiology ; prevention & control ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Prevalence ; Young Adult

Objective To effectively monitor the time trend of HIV prevalence and epidemic among injecting drug users ( IDUs ), female sex workers (FSWs), sexually transmitted disease (STD)clinic attendants, men who have sex with men (MSM) and pregnant women etc., to provide information for evaluation on strategies regarding comprehensive HIV prevention and effectiveness of intervention, in China. Methods Cross-sectional studies had been repeatedly conducted on sentinel surveillance program with uniformed questionnaire and collection of venous blood specimen among IDUs, FSWs, STD clinic attendants, MSM and pregnant women, from 1995 to 2009. Results HIV prevalence (median) among IDUs had been increasing rapidly since 1996, reaching the plateau (19.2%) in 1999-2000, then gradually decreasing in 2001-2003, and leveling off since 2004 in those relatively hard-hit epidemic provinces. HIV prevalence among drug users had increased a bit since 2001, achieving high level (2.9%) in 2004 before leveling off, showing a slight downward trend in the past two years among the medially-hit epidemic provinces, while it maintained at a low level in those low epidemic provinces. HIV prevalence (median) among FSWs had been slightly increasing,reaching a high level ( 1.5% ) in 1999, then decreased and maintained at a low level in relatively hard-hit provinces. HIV prevalence among FSWs maintained at a low level in medially-hit and low epidemic provinces. HIV prevalence(median) among STD clinic attendants had been maintained at a lower level, lower than 0.7%. HIV prevalence among MSM had been consistently greater than 1%, and was increasing year by year, becoming one of the most important drivers of the HIV/AIDS epidemic over the past years. HIV prevalence (median) among pregnant women had slightly been increasing since 1997, then leveling off over the past years in those relatively hard-hit provinces (0.3%). HIV prevalence among pregnant women had maintained at a low level in the medial and low epidemic provinces. Conclusion Data from sentinel surveillance programs showed a leveling-off situation on HIV prevalence rates among IDUs, FSWs, sexually transmitted infections clinic patients and pregnant women. However, HIV prevalence among MSM had increased significantly since 2005, indicating the high speed of transmission among this population. Epidemic among IDUs had still been concentrated in those relatively hard-hit epidemic provinces. As for FSWs and STD patients, the overall epidemic remained at low level, but the areas of epidemic kept expanding.

Objective To analyze the hepatitis C virus (HCV) prevalence among 8 populations in 2009-2012 in China,using the HIV sentinels surveillance data.Methods Cross-sectional studies had been repeatedly conducted on HIV Sentinel Surveillance with uniformed questionnaire and collection of venous blood specimen among 8 populations including drug users (DUs),men who have sex with men (MSM),female sex workers (FSW),male STD clinic attendees,male long distance truck drivers,male mobile populations,pregnant women and young students from April to June 2009 to 2012 on the yearly base.Blood testing for HIV,syphilis and HCV infections have been routinely performed on all participants of 8 populations recruited during the surveillance period.Results HCV prevalence among drug users had been relatively high,between 40.0％-43.0％ in 2009-2012.HCV prevalence of injection drug users (IDUs) showing a slight increasing trend in the past four years from 55.0％ in 2009 to 65.0％ in 2012.HCV prevalence rates among other 7 populations maintained low remained less than 1.0％ in the past three years.Conclusion Data from sentinel surveillance programs showed that HCV prevalence of DUs maintained high but the other 7 populations at low levels in the HIV sentinel surveillance areas in China.Blood-borne was the most common transmission pathways of HCV.IDU was one of the predominant models of HCV transmission in China and continued to fuel the HCV epidemics throughout the country.

Recently, suppressor of cytokine signaling-3 (SOCS3) has been shown to be an inducible endogenous negative regulator of Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway which is relevant in inflammatory response, while its functions in acute liver failure and HBV-induced acute-on-chronic liver failure (HBV-ACLF) have not been fully elucidated. In this study, we explored the role of SOCS3 in the development of mouse hepatitis virus strain 3 (MHV-3)-induced acute liver failure and its expression in liver and peripheral blood mononuclear cells (PBMCs) of patients with HBV-ACLF. Inflammation-related gene expression was detected by real-time PCR, immunohistochemistry and Western blotting. The correlation between SOCS3 level and liver injury was studied. Our results showed that the SOCS3 expression was significantly elevated in both the liver tissue and PBMCs from patients with HBV-ACLF compared to mild chronic hepatitis B (CHB). Moreover, a time course study showed that SOCS3 level was increased remarkably in the liver of BALB/cJ mice at 72 h post-infection. Pro-inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, were also increased significantly at 72 h post-infection. There was a close correlation between hepatic SOCS3 level and IL-6, and the severity of liver injury defined by alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, respectively. These data suggested that SOCS3 may play a pivotal role in the pathogenesis of MHV-3-induced acute liver failure and HBV-ACLF.

Objective:To investigate the expression of urinary exosomal miR-150-5p in different age groups and its relationship with kidney aging.Methods:Seventy-six subjects were recruited at the geriatrics department of Beijing Friendship Hospital from January 2014 to December 2016, with 15 in the young group, 35 in the middle-aged group, and 26 in the elderly group.We detected the expression of urinary exosomal miR-150-5p by real-time PCR from urine samples of different age groups and used Logistic regression analysis to evaluate the correlation of different levels of miR-150-5p with age, gender, history of hypertension, history of coronary heart disease, and hyperlipidemia.Additionally after overexpression and knockdown of miR-150-5p in human renal tubular cells of the HK2 cell line, we analyzed the expression levels of epithelial-to-mesenchymal transition(EMT)-related proteins by Western blotting.Results:Logistic regression analysis showed that age was closely related to estimated urinary exosomal miR-150-5p levels( P<0.05), and estimated urinary exosomal miR-150-5p levels in the elderly group(1.33±0.41)were significantly higher than those in the young group(0.88±0.40)( P<0.05). In addition, estimated urinary exosomal miR-150-5p levels became inversely proportional to the glomerular filtration rate as age increased. In vitro experiments showed that the expression levels of ZO-1 and E-cadherin in epithelial cells were slightly decreased after knockdown of miR-150-5p in renal tubular epithelial cells of the HK-2 cell line. Conclusions:The expression of miR-150-5p in urinary exosomes may be slightly increased in the elderly.It could inhibit the occurrence of renal EMT, and its expression is correlated with renal function.Mir-150-5p may also affect the expression of adhesion molecule-related proteins in HK-2 cells.MiR-150-5p in urinary exosomes is expected to become one of the genetic markers associated with renal aging.

BACKGROUNDNow lung volume reduction surgery (LVRS) has become one of the most effective methods for the management of some cases of severe chronic obstructive pulmonary disease (COPD). We evaluated the mid-term effects of LVRS on pulmonary function in patients with severe COPD.

METHODSTen male patients with severe COPD aged 38 - 70 years underwent LVRS and their pulmonary function was assessed before, 3 months and 3 years after surgery. The spirometric and gas exchange parameters included residual volume, total lung capacity, inspiratory capacity, forced vital capacity, forced expiratory volume in one second, diffusion capacity for CO, and arterial blood gas. A 6-minute walk distance (6MWD) test was performed.

RESULTSAs to preoperative assessment, most spirometric parameters and 6MWD were significantly improved after 3 months and slightly 3 years after LVRS. Gas exchange parameters were significantly improved 3 months after surgery, but returned to the preoperative levels after 3 years.

CONCLUSIONSLVRS may significantly improve pulmonary function in patients with severe COPD indicating for LVRS. Mid-term pulmonary function 3 years after surgery can be decreased to the level at 3 months after surgery. Three years after LVRS, lung volume and pulmonary ventilation function can be significantly improved, but the improvement in gas exchange function was not significant.

Adult ; Aged ; Exercise Tolerance ; Humans ; Lung Volume Measurements ; Male ; Middle Aged ; Pneumonectomy ; adverse effects ; methods ; Pulmonary Disease, Chronic Obstructive ; pathology ; physiopathology ; surgery ; Respiratory Function Tests ; Time Factors