Objective To compare and evaluate two type-specific primers PCR genotyping methods of hepatitis B virus ( HBV) which were established by Naito et al ( Naito method) and our lab (new method). Methods The two genotyping methods were applied for detecting the plasmids containing the HBV genomes of genotype A or D or subgenotype B1 or C2 and the plasmids mixed with different proportion of subgenotypes B1 and C2. In addition, the genotypes of 113 serum samples of patients with chronic HBV infection from Shenzhen, Handan and Urumqi cities of China were identified by the two methods, respectively. The results were verified by PCR product based sequencing. Results The sensitivity of the two methods showed no difference when they were applied to detect the plasmids containing the HBV genomes of genotype A or D or subgenotype B1 or C2. While detecting the plasmids mixed with different proportion of subgenotypes B1 and C2, the sensitivity of the new method was superior than that of Naito method. Meanwhile, the specificity of the new method was obviously superior than that of Naito method. Both of the two methods were highly sensitive in identification of HBV genotypes of serum samples with a single genotype. However, the new method showed more sensitive in identification of the B/C mix strains than that of Naito method. The total coincidence rate between the two methods was 83. 2% (94/113), with the discrepancy of 16. 8% (19/113). Fifteen of the 19 inconsistent genotypes by the two methods were selected and their PCR products were sequenced directly. The sequencing results were identical with that of the new methods, but not with that of the Naito method. Conclusion The new method is more sensitive, and its specificity is superior to the Naito method. It could be used for clinical and epidemiological studies on HBV genotype and subgenotype in China.

Bone marrow mesenchymal stem cells (MSCs), multipotent stem cells, can replicate as undifferentiated cells and have the potential to differentiate into different lineages of mesenchymal tissues, including bone, cartilage,endothelial, neural, smooth muscle, skeletal myoblasts, and cardiac myocyte cells. The ischemia-induced death of cardiomyocytes results in scar formation and reduced contractility of the ventricle. Several preclinical and clinical studies have supported the notion that MSCs therapy may be used for cardiac regeneration.When transplanted into the infracted heart, MSCs prevent deleterious remodeling and improve recovery, but the mechanism is not clear. In this work,we review evidence and new prospects that support the use of MSCs in cardiomyoplasty.

Objective To establish a hepatitis B virus (HBV) nested PCR (nPCR) for detection of genotypes A-D and subgenotypes B1,B2, C1 and C2. Methods The entire HBV nucleotide sequences of genotypes A-H retrieved from GenBank were compared and analyzed by DNAStar software. The PCR primers were designed by Primer Premier 5.0 software,and the nPCR for genotyping HBV/A-D as well as subgenotyping B1, B2,C1 and C2 were established. There were 3 steps in the process:step 1 for genotypes B, D and subgenotypes C1, C2 with the amplification of Mix A; step 2 for genotype A with the amplification of Mix B; step 3 for subgenotypes B1 and B2 with the amplification of Mix C in the second-ound PCR, based on the first-round amplification procedure. A total of 68 serum samples from patients with chronic HBV infection were detected by nPCR. 15 of 68 sera were selected randomly and their PCR products were directly sequenced to confirm the accuracy of the method. Results Among 68 serum samples of patients with chronic HBV infection detected by the nPCR, 23.53% (16/68) were infected with B2, 11.76% (8/68) with C1,48.53% (33/68) with C2,1.47% (1/68) with D,11.76% (8/68) with B2C2 mix strains,1.47% (1/68) with C2D mix strains and 1.47% (1/68) with B2/C1/D mix strains. The sequencing analysis of the 15 serum samples had the same results as detected by nPCR. Conclusion nPCR is a simple,rapid method and able to detect genotypes A-D and subgenotypes B1 ,B2 ,C1 and C2 subtypes of HBV with both high sensitivity and specificity.

In order to evaluate the immunogenicity of HPV 58 L1 DNA vaccines, five DNA vaccines had been constructed with pcDNA3.1 vector containing different L1 genes of HPV 58, which were designated as L1h, L1hDeltac, L1S, L1SM and L1wt. The protein expression of DNA vaccines in vitro was tested by Western blot. The ability of forming pseudovirus was evaluated by transfecting DNA vaccine together with pcDNA3.1-h58L2 and pcDNA3.1-GFP into 293FT cells. The neutralizing antibodies and cellular immune response produced in BALB/c mice immunized with the DNA vaccines were detected by using pseudovirus-based neutralization assay and ELISPOT respectively. The results showed that the five DNA vaccines had been successfully constructed; the level of protein expression of L1hDeltac was the highest and those for L1S and L1SM were of medium, while no expressed target protein of L1wt was detected. Only L1S could form the pseudovirus while the other four vaccines could not. L1S and L1h could induce neutralizing antibody. However, the average titer of neutralizing antibody for L1S (1:6,400) was much higher than that for L1h (1:48) and the other three vaccines could not induce neutralizing antibody. No cellular immune response for all five DNA vaccines was detectable by ELISPOT. The results indicated that DNA vaccine against HPV 58 can form pseudovirus in vitro, also can induce high level of neutralizing antibodies. This provides reference for screening HPV vaccine in future.
Animals ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Female ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Models, Genetic ; Neutralization Tests ; Papillomaviridae ; immunology ; Papillomavirus Vaccines ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vaccines, DNA ; genetics ; immunology

Objective To evaluate the preventive and therapeutic effect of oral solution of Niao Du Kang (尿毒康) on acute renal tubular necrosis (ATN) in Sprague-Dawley (SD) rats,and preliminarily approach its mechanisms.Methods The oral solution was composed of traditional Chinese medicinal herbs of Radix et Rhizoma Rhei (大黄),Radix Sanguisorbae Offieinalis (地榆),Radix Salviae Miltiorrhizae (丹参), Radix Astragali seu Hedysari (黄芪),Flos Carthami Tinctorii (红花) and so on.Sixty-four SD male rats were randomly divided into 4 groups:normal control group,ATN model group,verapamil treatment group and traditional Chinese medicine,oral solution of Niao Du Kang treatment group,every group having 16 rats.. The ATN model of SD rats was induced by intramuscular injection of 50% glycerin mixed with isotonic saline solution at the back of bilateral lower extremities.After the model was established in each group ,the levels of blood urea nitrogen (BUN),levels of blood serum creatinine (SCr),renal failure index (RFI) and renal pathological changes were detected at different time points after modeling for 12 and 24 hours,and the therapeutic effect of oral solution of Niao Du Kang was observed.Results After the model establishment, Niao Du Kang oral solution could lower the elevation of blood BUN,SCr,RFI in the model,and compared with the normal control group,there were statistical significances (all P

OBJECTIVETo observe the effect of rhynchophylline on methamphetamine-dependent zebrafish and explore the possible mechanism.

METHODSZebrafish were divided into control group, amphetamine group, low- (50 mg/kg) and high (100 mg/kg)-dose rhynchophylline groups, and ketamine (150 mg/kg) group. Conditioned place preference (CPP) was induced in zebrafish with methamphetamine, and the staying time in the drug box and the tracking map of the zebrafish were observed with Noldus Ethovision XT system. The protein expressions of TH, NR2B and GLUR2 in the brain of zebrafish with CPP were detected with Western blotting.

RESULTSCompared with the control group, zebrafish in methamphetamine group showed significant variations in the staying time and swimming distance in the drug box after conditioning (P<0.05) with obvious alterations of NR2B, TH and GLUR2 expressions in the brain (P<0.05). Treatment of methamphetamine-dependent zebrafish with high-dose rhynchophylline significantly reduced the variations in the staying time and swimming distance in the drug box (P<0.05) and in the expressions of NR2B, TH and GLUR2 in the brain (P<0.05).

CONCLUSIONRhynchophylline can inhibit methamphetamine dependence in zebrafish, the mechanism of which may involve the expressions of TH, NR2B and GLUR2 proteins in the brain.

To find out whether the mutations of HIV-1 Env have influence on the assembly of pseudovirus and their abilities to infect cells, site-directed mutation (A457D)was performed using cycling mutagenesis and selection of mutants with DpnI. Transformation and plasmid purification technologies were used to obtain mutated env clone. Then both the prototype and the mutant were co-transfected with pSG3(delta(env)) to 293FT cells, respectively. Single-cycle infection assay was employed to analyze the effect of the prototype and the mutant on the ability of functional pseudovirus assembly. The transient expression of both the prototype S12-42-1 and mutant S12-42M were confirmed by Western blot essay. The S/CO value was less than 1 for S12-42-1 and 6.65 for S12-42M, demonstrating the functional pseudovirus was generated only for S12-42M. So mutation on HIV-1 Env has influence on the assembly of pseudovirus and their abilities to infect cells.
Amino Acid Sequence ; Base Sequence ; Cell Line ; HIV Infections ; virology ; HIV-1 ; chemistry ; genetics ; physiology ; Humans ; Molecular Sequence Data ; Mutation ; Sequence Alignment ; Virion ; genetics ; physiology ; Virus Assembly ; env Gene Products, Human Immunodeficiency Virus ; chemistry ; genetics ; metabolism

Although mesenchymal stem cells (MSCs) are increasingly used to treat graft-versus-host disease (GVHD), their immune regulatory mechanism in the process is elusive. The present study aimed to investigate the curative effect of third-party umbilical cord blood-derived human MSCs (UCB-hMSCs) on GVHD patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and their immune regulatory mechanism. Twenty-four refractory GVHD patients after allo-HSCT were treated with UCB-hMSCs. Immune cells including T lymphocyte subsets, NK cells, Treg cells and dendritic cells (DCs) and cytokines including interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-α) were monitored before and after MSCs transfusion. The results showed that the symptoms of GVHD were alleviated significantly without increased relapse of primary disease and transplant-related complications after MSCs transfusion. The number of CD3(+), CD3(+)CD4(+) and CD3(+)CD8(+) cells decreased significantly, and that of NK cells remained unchanged, whereas the number of CD4(+) and CD8(+) Tregs increased and reached a peak at 4 weeks; the number of mature DCs, and the levels of TNF-α and IL-17 decreased and reached a trough at 2 weeks. It was concluded that MSCs ameliorate GVHD and spare GVL effect via immunoregulations.
Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; methods ; Cytokines ; metabolism ; Dendritic Cells ; metabolism ; Female ; Graft vs Host Disease ; immunology ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunomodulation ; Killer Cells, Natural ; metabolism ; Male ; T-Lymphocyte Subsets ; metabolism ; Transplantation, Homologous ; adverse effects ; Young Adult

Objective To evaluate the feasibility,reliability,validity and responsiveness of a Chinese Menopause Rating Scale (CMRS).Methods Cross-sectional survey and convenience sampling were adopted. Participants:women with menopause syndrome and those in menopause but without menopause syndrome were recruited.All participants were asked to complete the CMRS,Kupperman Index,WHOQOL-BREF and MENQOL.The Self-control observation design was adopted when the responsiveness was evaluated.Patients were treated with TCM for weeks.MRSTCM was evaluated before and after the treatment.Results (1) Feasibility:3343 participants including 2320 patients and 1023 menopause women,were surveyed in 8 different settings.The recovery rate of CMRS was 100%,with a response rate as 99.7%.The completion of the CMRS took 10.30 minutes on average.(2)Reliability:Cronbach's alpha of CMRS,soma dimension,psychology dimension and community dimension of CMRS were 0.93,0.87,0.89 and 0.73 respectively,with the correlation coefficient of split half of the CMRS.Soma dimension,psychology dimension and community dimension were 0.92,0.89,0.86 and 0.73 respectively and the test-retest correlation coefficient of MRSTCM,the soma dimension,psychology dimension and community dimension were as 0.88,0.91,0.85 and 0.77 respectively.(3) Validity:CMRS was established on the basis of connotation of menopause syndrome,and a series of steps were adopted to modify the scale.CMRS was applicable for patients with menopause syndrome.CMRS seemed to have had good content-related validity.The result of exploratory factor analysis was accorded with the theory frame of CMRS by and large.The correlations between CMRS and KI,CMRS and WHOQOLBREF,CMRS and MENQOL seemed good.The CMRS was able to discriminate between groups of people with or without menopausal syndrome and bad good discriminative validity.(4) Responsibility:The CMRS was measured based on 174 patients with menopausal syndrome before and after the TCM therapy.Our result showed that the CMRS having the ability to measure the clinically important differences.Conclusion CMRS was suitable for outcome assessment of menopausal syndrome.This primary research proved that the CMRS had good feasibility,reliability,validity as well as responsiveness.

OBJECTIVETo explore the relationship between distorted weight perception and suicide ideation among normal weight adolescents in Guangdong province.

METHODSThis study used the data of Guangdong Provincial Youth Health Risk Behavior Survey in 2004 and 2007. To identify the association between distorted weight perception and suicide ideation, a logistic regression analysis was performed. The effects of age, economic status, mother's education, mental health, and depression were also adjusted.

RESULTSA total of 12 729 people participated in this study, 6096 males and 6633 females. The prevalence of suicide ideation among normal weight adolescents was 13.58% (1729/12 729) in the past 12 months, with higher prevalence in girls (16.15%, 1071/6633) than that in boys (10.79%, 658/6096) (χ(2) = 77.71, P = 0.00). It was common that the adolescents misperceived their weight. Only 44.93% (5719/12 729) of normal weight students correctly perceived their body weight while 43.52% (5540/12 729) of them overestimated their weight and 11.43% (1455/12 729) underestimated their weight. The distorted weight perception in girls (65.58%, 4350/6633) was higher than that in boys (43.39%, 2645/6096) (χ(2) = 993.91, P = 0.00). Distorted weight perception was significantly associated with suicide ideation after controlling for factors age, economic status, mother's education, mental health, and depression. The students who overestimate their body weight were more likely to have suicide ideation than that who correctly perceived their weight (OR = 1.43, 95%CI: 1.27 - 1.61).

CONCLUSIONThe prevalence of distorted weight perception was high and it significantly associated with suicide ideation.

Adolescent ; Body Weight ; Female ; Humans ; Male ; Self Concept ; Students ; psychology ; Suicide, Attempted ; psychology ; statistics & numerical data ; Surveys and Questionnaires