This study aimed to amplify major genome segments (VP7, VP4, VP6, VP2 and NSP2-5) of porcine G3 group A rotavirus (GARV) LLZ212 isolated in our laboratory, determine their genotypes, and explore the evolutionary relationships between G3 GARV strains isolated from humans and pigs in Lulong County, Hebei Province, China. Major genome segments of seven GARV strains were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the segments were sequenced. The genome segments of seven GARV strains were determined by the online RotaC genotyping tool (RotaC v2.0). The reference sequences of each GARV genome segment were downloaded from GenBank. Homology and phylogenetic evolutionary analyses were conducted using the MEGA 5.0 and DNAStar software packages. LLZ212 isolated from pigs in Lulong had the following genotype: G3-P[8]-I5-C1-N1-T1-E1-H1. All human GARV strains had the following genotype: G3-P[8]-I1-C1-N1-T1-E1-H1. The VP7, VP4, NSP4 and NSP5 genes of the LLZ212 strain had the highest nucleotide identities with the human GARV E885, CMH014/07, Wa and RMC321 strains, respectively, and these clustered together in a sublineage. The VP6, NSP4 and NSP5 genes of the LLZ212 strain shared the highest nucleotide identities with the porcine GARV PRG921 strain, while VP2 associated most closely with porcine GARV OSU strain, and these also clustered in a sublineage. A rare porcine G3-P[8]-I5-C1-N1-T1-E1-H1 GARV strain was identified, which may represent a reassortment between porcine and human viruses. In conclusion, the VP7, VP4, NSP4 and NSP5 genes of LLZ212 share high levels of sequence identity with human GARV, while VP2, VP6, NSP2 and NSP3 cluster with porcine GARV.
Animals ; Capsid Proteins ; genetics ; Child, Preschool ; China ; epidemiology ; Evolution, Molecular ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; veterinary ; virology ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Nonstructural Proteins ; genetics

Objective To determine whether regulatory T cells(Tr)are increased in patients with tuberculosis and whether they are associated with its immunopathology.Meantime,to investigate the possibility of tuberculosis(TB)as a model for studying Tr functions.Methods The lymphocyte subsets were isolated from peripheral blood mononuclear cells by sorting with flow cytometry.Total cellular RNA was extracted and RT-PCR was performed to detect the Foxp3 mRNA in purified CD3+CIM+T cells,CD3+CD8+T cells and non-CD3+CD4+CD8+T cells.Using FACS analysis.we further investigated the distribution of Foxp3+ population in CD4+ CD25+T cells.Finally,we compared the percentage of CD4+CD25highFoxp3+T cells present in 51 active patients with tuberculosis and 40 uninfected healthy control subjects by FACS.The detection of Tr infiltration of Foxp3+ cells were performed with immunohistochemistry(IHC)method on tuberculosis pathological sections.Results Foxp3 was specific expressed in CD3+CD4+T cells,either in tuberculosis patients or healthy control subjects.Foxp3+ T cells took about 85%fraction of CD4+ CD25highpopulation.We used CD4+CD25high Foxp3+as a detective markers for Tr in the FACS analysis.The results showed that patients with active TB had a 4.4 fold higher percentage within the CD4+T cells in peripheral blood compared to healthy control group(modian,1.01%vs 0.23%,P<0.01).Much higher frequency of Tr were found along with T cells infiltration at the tuberculosis pathological tissues.A few individuals that we can followed indicated the expanded Tr was declined after curative treatment with operation.Conclusion Tr cells are increased in tuberculosis patients and closely correlate with its immunopathology.Tuberculosis should be a valuable model for Tr functional study.

This study was aimed to investigate the chimerism and fusion gene expression in patients with CML after allo-HSCT, to analyse engraftment and minimal residual disease by using STR-PCR combined with RT-PCR qualitative and quantitative assays, and to evaluate their clinical value for predicting disease relapse. 4 relapsed patients with CML after allo-HSCT were dynamically investigated. Qualitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was performed by RT-PCR. The results showed that the 100% donor chimerism appeared in 4 patients on day 28 after transplantation and bcr/abl expression was negative, but the 4 patients were in status of unstable mixed chimerism (DC: 0% - 80.4%) at the different time points during the following up with bcr/abl gene positive. 2 patients of them were continuously mixed chimerism after relapse of CML, the other 2 changed from MC to CC by intervention of clinical treatment. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and CML relapse, and bcr/abl gene expression was positive. It is concluded that the results of STR-PCR in the range of its sensitivity fully correspond with bcr/abl tests in patients. The combination of STR-PCR with RT-PCR will provide a highly sensitive and valuable tool for evaluating engraftment, graft rejection, and relapse and predicting GVHD. Furthermore, it can provide a basis for early intervention of clinical treatment, and can identify these high risk patients with molecular or cytogenetic relapse after allo-HSCT.
Fusion Proteins, bcr-abl ; genetics ; metabolism ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; therapy ; Neoplasm Recurrence, Local ; genetics ; Neoplasm, Residual ; diagnosis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Transplantation Chimera ; Transplantation, Homologous

BACKGROUNDPulmonary embolism (PE) can be difficult to diagnose in elderly patients because of the coexistent diseases and the combination of drugs that they have taken. We aimed to compare the clinical diagnostic values of the Wells score, the revised Geneva score and each of them combined with D-dimer for suspected PE in elderly patients.

METHODSThree hundred and thirty-six patients who were admitted for suspected PE were enrolled retrospectively and divided into two groups based on age (≥65 or <65 years old). The Wells and revised Geneva scores were applied to evaluate the clinical probability of PE, and the positive predictive values of both scores were calculated using computed tomography pulmonary arteriography as a gold standard; overall accuracy was evaluated by the area under the curve (AUC) of receiver operator characteristic curve; the negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were calculated.

RESULTSNinety-six cases (28.6%) were definitely diagnosed as PE among 336 cases, among them 56 cases (58.3%) were ≥65 years old. The positive predictive values of Wells and revised Geneva scores were 65.8% and 32.4%, respectively (P < 0.05) in the elderly patients; the AUC for the Wells score and the revised Geneva score in elderly was 0.682 (95% confidence interval [CI]: 0.612-0.746) and 0.655 (95% CI: 0.584-0.722), respectively (P = 0.389). The negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were 93.7%, 100%, and 100% in the elderly, respectively.

CONCLUSIONSThe diagnostic value of the Wells score was higher than the revised Geneva score for the elderly cases with suspected PE. The combination of either the Wells score or the revised Geneva score with a normal D-dimer concentration is a safe strategy to rule out PE.

Aged ; Aged, 80 and over ; Angiography ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Pulmonary Embolism ; diagnosis ; diagnostic imaging ; metabolism ; Retrospective Studies

OBJECTIVETo introduce practical diagnostic criterion of blood stasis syndrome (BSS), and to evaluate its reliability and validity.

METHODSBy referring to three diagnostic criteria of BSS [practical diagnostic criterion of BSS (criterion A), diagnostic criterion of BSS in 1986 (criterion B), Consensus of Integrative Medicine on BSS Diagnosis in 2011 (criterion C)], 712 patients from different departments of Xiyuan Hospital were recruited. The reliability of criterion A and its consistency with the other two criteria were assessed using Kappa coefficient. A Bayesian approach was also employed to assess the sensitivity and specificity of criterion A.

RESULTSAccording to the consistency check, criterion A presented good consistency when used by different researchers (the diagnostic accordance rate was 91. 96%, Kappa =0. 82, P <0.001). Meanwhile, there was an acceptable diagnostic consistency among the three diagnostic criteria. Bayesian estimation suggested that criterion A had higher sensitivity but similar specificity, as compared with criterion B or criterion C. Compared with criterion B [the median of sensitivity and specificity were 0. 762 (95% Cl: 0. 731 -0. 790) and 0. 902 (95% Cl: 0. 858 -0. 936) respectively, the median of sensitivity and specificity of criterion A were 0. 911 (95% CI: 0. 888 - 0. 930) and 0. 875 (95% CI: 0. 826 - 0. 915) respectively. Estimating the difference between criterion A and B, the median of sensitivity and specificity were 0. 149 (95% CI: 0. 112 -0.184) and -0. 026 (95% CI:-0. 085 -0. 033) respectively. Compared with criterion C [the median of sensitivity and specificity were 0. 831 (95% Cl: 0. 804 -0. 857) and 0. 892 (95% CI: 0. 848 - 0. 926) respectively], the median of sensitivity and specificity of criterion A were 0. 912 (95% CI: 0. 889 -0. 932) and 0. 880 (95%CI: 0. 833 - 0.919) respectively. Estimating the difference between criterion A and C, the median of sensitivity and specificity were 0. 081 (95% CI: 0.047 - 0.114) and -0.011 (95%CI: -0.070 -0.046) respectively.

CONCLUSIONCompared with criterion B and C, criterion A not only had better reliability, but also could significantly improve the sensitivity without obviously lowering the specificity.

Bayes Theorem ; Consensus ; Hematologic Diseases ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Reproducibility of Results ; Sensitivity and Specificity

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.
China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Molecular Sequence Data ; Parvoviridae Infections ; epidemiology ; virology ; Phylogeny ; Seasons

OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.

METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.

RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.

CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.

Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea

Abstract:This study aims to investigate the genetic characteristics of group A rotavirus (GARV) G9P[8] strains from infantile diarrhea samples in Hebei Lulong region from 2009 to 2011. We randomly selected five GARV G9P[8] strains in Hebei Lulong region from 2009 to 2011, amplified the 11 gene fragments of GARVs by RT-PCR, and analyz their full-genome sequences by homology and phylogenetic analysis with DNAStar and MEGA. The nucleotide homology between strains LL11131077 and LL11131083 in 2011 was significantly higher than hat etween them and the other three strains in 2009 and 2010. The G9P[8] GARVs circulating in Hebei Lulong region from 2009 to 2011 elenged to the same genotype as the prevalent G9P[8] GARVs in other parts of the world. However,the two strains in 2011, compared with those in 2009 and 2010, were located in a different sub-branch of the phylogenetic tree and had amino acid mutations at many sites.
China ; Feces ; virology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Viral Proteins ; genetics

Objective To investigate the effect of the Gardenia extracts T9 on herpes simplex virus (HSV) viral protein 16(VP16) in HSV-1 infected mice's brains and its potential anti-viral mechanism. Methods HSV-1 infection was induced in BALB/c mice, the expression of HSV VP16 was detected by RTPCR on the 4th, 7th, 10th, 14th, and 21st day. Results The expression of VP16 mRNA decreased in both groups of large and low doses compared with virus group at the same time. Conclusion Gardenia extracts T9 can down-regulate the expression of VP16 mRNA in HSV infected mice's brains, and it may be one of its anti-HSV mechanisms.

ObjectiveTo study the value of liquid-based cytology and colposcopy in screening for cervical lesion among urban community women of Beijing.MethodsA total of 795 women aged 20～54 years with sexual activity living in Zhanlanlu Community of Beijing were screened for cervical lesion Cervcal specimen was collected for thin-layer,liquid-based cytology test (LCT) from each of the participants in gynecologic examination.Colposcopy and biopsy were performed for the women with positive LCT.ResultsForty-five of 795 (5.7%) women were positive for LCT[≥ASC-US (atypical squamouscell of undetermined significance)],with 33 of ASC-us,eight of low-grade squamous intraepithelial lesion (LSIL),three of high-grade squamous intraepithelial lesion (HSIL),one of atypical glandular cells (AGC).Five of 45 women with positive LCT refused to accept colposcopy.Among 40 women with colposcopy and biopsy,chronic cervicitis was diagnosed in 11(27.5%),cervical condyloma acurninatum in 14(35.0%),cervical intraepithelial neoplasia (CIN) 1 in seven(17.5%),CIN 2 in three (7.5%),CIN 3 in four(10.0%),and early invasive cervical cancer in one(2.5%).In 750 women with negative LCT,cervical condyloma acuminature was diagnosed in two(0.3%),CIN 1 in five(0.7%)and low-grade glandular intraepithelial lesion (LGIL) bin one(0.1%).Sensitivity and specificity of LCT screening for cervical lesion(≥CIN 1)were 71.4%and 94.2%,respectively,with positive and negative predictive values of 37.5%and 99.2%.respectively,and those screening for cervical lesion more than CIN 2 were 100.0%, 96.0%,20.5%and 100.0%,respectively.ConclusionsMore attention should be paid to early screening for cervical lesion in urban community women.LCT combined with colposcopy and biopsy provide very helpful information in screening for early cervical cancer.