To determine the etiologic agent of an outbreak of influenza viruses from Changsha Foothill Mountain International School in 2009, and to analyze the HA Gene Characteristic of the H1N1 influenza viruses. Twenty-five nasopharyngeal swab specimens from the outbreak of influenza viruses were tested by conventional RT-PCR and influenza viruses isolated simultaneously. Virus isolated (A/Yuelu/314/2009) from the outbreak was sequenced by CEQTM 8000 Genetic Analysis System and the sequencing results submitted to GenBank (Accession No: FJ912843), then the sequencing data was analyzed by ClustalX and Mega4.1softwares. Results showed the influenza viruses A(H1N1) of positive were 18 cases by influenza viruses isolated tests and 21 cases by conventional RT-PCR, respectively. The nucleotide and amino acid sequence homology of the HA gene of A/Yuelu/314/2009 are 99% compare with the vaccine strain (A/Brisbane/59/2007) in 2008~2009 years. The HA sequence data also showed that had 6 amino acid mutations (V148A, S158N, G202A, I203D, A206T, W435R), and the S158N located at antigenic site B of HA protein. Nine potential glycosylation sites (27, 28, 40, 71, 151, 176, 303, 497, 536) in the HA sequence of A/Yuelu/314/2009 is the same with A/Brisbane/59/2007, and the sequences of potential glycosylation sites were conserved. In this study, laboratory evidence diagnosed seasonal influenza A virus (H1N1) as the etiologic agent of the outbreak. The virus isolated (A/Yuelu/314/2009) strain of H1N1 subtype is not a new variant in Changsha in 2009 compare with the vaccine strain (A/Brisbane/59/2007), the outbreak of influenza A virus (H1N1) from Changsha Foothill Mountain International School maybe are caused by the change in genetic characteristics between vaccine strains and the decreased of immunity to influenza A virus (H1N1) in the crowd.

Objective To investigate the efficacy of the bioactive artificial vertebrae of a nano- hydroapatite crystals and polyamide 66 composite(n-HA/PA66)to restore the height and architecture of thoracolumbar burst fracture.Methods From December 2003 to February 2006,38 patients(29 males and 9 females)with a mean age of 35.6 years(17-63 years)were treated surgically through anterior ap- proach for decompression and implanted with the bioactive artificial vertebrae of n-HA/PA66 composite to reconstruct the structure of the thoracolumbar burst fractured vertebra.Results All the patients were successfuly followed-up for an average of 8 months,ranging from 6 to 21 months.The bioaetive artificial vertebrac of n-HA/PA66 composite were fused with the receptor bone 3-4 months after operation.The neu- rological function of the patients was restored partially or completely.The thoracolumbar spine was stable during physical examination and the height of thoraeolumbar burst fractured vertebrae that had been restored did not changa during the follow-up.Conclusions Our results show the bioaetive artificial vertebrae of n-HA/PA66 can restore the height and structure of thoracolumbar burst fractured vertebrae and reconstruct the structure of the tboraeolumbar vertebrae effectively,indicating that the bioaetive artificial vertebrae of n- HA/PA66 can be used extensively in clinical spinal surgery.

OBJECTIVETo observe the effect of Gansui Banxia Tang plus-minus Gansui and Gancao anti-drug combination on hepatic and renal functions in malignant ascites rats to explore whether the efficacy or toxicity associated with the anti-drug combination.

METHODThe male wistar rats were randomly divided into a blank group, model group, furosemide group, Gansui Banxia Tang group, Gansui Banxia Tang removed Zhigancao group, Gansui Banxia Tang removed Cugansui group, Gansui Banxia Tang removed Zhigancao and Cugansui group. In addition to normal feeding, every morning except for the blank group and model group, the rest of the group was given drugs, the control group and the model group was given distilled water, the volume is 10 mL x kg(-1). Administered five days, all rats were fasted but except water for 24 hours to collect urine. Administered nine days all rats were fasted but except water for 12 hours, we need to weigh weight of rats. When we remove the ascites, we also need to weigh weight of rats. We use the weight before removing ascites minus weight after removing ascites to indirectly measure the amount of ascites. When we remove the ascites, we need to abdominal aortic blood, centrifuge testing renin, angiotensin II, aldosterone, antidiuretic hormone and other indicators.

RESULTThe effect of Gansui Banixa Tang on increasing the net weight, lowering abdominal circumference and body weight ratio, lowering renin, angiotensin, aldosterone, antidiuretic hormone is better than the other treatment group.

CONCLUSIONIn diuresis party, the group of Gansui Banxia Tang is better than the group of Gansui Banxia Tang remove Zhigancao or Cugansui or Zhigancao and Cugansui, renin-angiotensin-aldosterone system may play a diuretic effect of its one way.

Aldosterone ; metabolism ; Angiotensin II ; metabolism ; Animals ; Ascites ; drug therapy ; metabolism ; physiopathology ; Body Weight ; drug effects ; Diuretics ; pharmacology ; therapeutic use ; Dose-Response Relationship, Drug ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Renin-Angiotensin System ; drug effects

To develop a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Norwalk GII. 4 primers which recognized 6 distinct regions on the RNA-dependent RNA polymerase gene of Norwalk GII were designed and used for LAMP assay. Norwalk GII RNA was amplified under isothermal conditions (65 degrees C) for 120 min, and LAMP results were then judged with naked eye, SYBR Green I staining, electrophoretic analysis and restriction digestion. To evaluate the specificity of the RT-LAMP, 48 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses were tested. To compare the sensitivity of the RT-LAMP with that of conventional RT-PCR, Norwalk GII RNA was serially diluted and amplified by RT-LAMP and RT-PCR, respectively. With 46 fecal specimens of Norwalk GII, observation with naked eyes, SYBR Green I staining and electrophoretic analysis were able to detect the PCR products in the RT-LAMP assay. The specificity of RT-LAMP products was also confirmed by digestion of the RT-LAMP products with restriction enzymes. No RNA amplification was observed in 2 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses. The specificity of the RT-LAMP assay with regard to RT-PCR were 100% for Norwalk GII. The detection limits of RT-LAMP was 15.6 pg/tube for Norwalk GII and similar to that of a RT-PCR assay. Compared to RT-PCR, the RT-LAMP assay has been proven to be a rapid, sensitive, specific and accurate method for detection of the Norwalk GII in fecal specimens, and that RT-LAMP assay is potentially useful for the rapid detection of Norwalk GII from fecal specimens in outbreaks of infectious diarrhea.
Caliciviridae Infections ; virology ; Feces ; virology ; Humans ; Norwalk virus ; genetics ; isolation & purification ; RNA Replicase ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics

Child ; China ; epidemiology ; Genes, Viral ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology

Objective To investigate the risk of H5Nl subtype avian influenza virus (AIV)transmission in the poultry market environment in Changsha city.H5N1 antibody levels among the groups related occupational exposure and AIV nucleic acid in the environment of poultry markets were detected.The characteristics of hamagglutinin (HA) genes of H5N1 AIV in the environment were analyzed.Methods One district and one county from Changsha city were selected randomly and two poultry markets at inner city or township levels were selected in the same district or county respectively.H5N1 antibody of the occupational exposure groups in the poultry market was tested and AIV nucleic acid in the poultry market environnent monitored.One hundred and two blood samples of the occupational exposure groups were tested for H5N1 antibody with single radioimmunoassay diffusion hemolysis (SRH) while 160 environment samples(from sewage,birds stools,feathers and smearing samples of poultry cages) in the poultry market were also detected for AIV nucleic acid with real-time PCR method.Four sewage samples of H5N1 subtype AIV were collected from poultry markets in Changsha,and the HA genes of H5N1 subtype AIV amplified by RT-PCR and then sequenced with TA cloning.Amino acid sequence alignment and phylogenetic tree analysis were conducted by Lasergene and Mega 5.0 software.Results The results through H5N1 antibody monitoring program showed that H5N1 antibody positive rates from workers were 25.5％ (26/102),50.0％ (9/18) and 25.4％ (17/67) respectively in the poultry markets of township and inner cities.H5N1 antibody positive rate in the township poultry markets was higher than in the inner cities poultry markets.Results from the surveillance on AIV nucleic acid showed that the overall H5 subtype positive rate in Changsha poultry markets was 31.3％ (50/160),and the positive rate of townships poultry markets was 37.3％ (31/83 ),which were both higher than those from the inner cities poultry markets (24.7％,19/77).H5 subtype AIV positive rate was different in the tested specimens,with ranking of positive rates were sewage (50.0％,24/48),feathers (44.5％,4/9 ),birds stools ( 29.8％,14/47) and smearing samples of poultry cages (14.3％,8/56),with statistically significant differences (P＜0.01).Four H5N1 HA genes TA cloning were successfully constructed and identified as Eurasian branch,similar to viruses isolated in mainland China and Hong Kong in the same group,according to genetic analysis.Sequence data of the four HA genes showed the same feature of high pathogenicity,compared to the H5N 1 AIV from mainland China of human origin.The receptor specificities were still with avian influenza origin (QSG)and the connecting peptide between HA1 and HA2 possessing the polybasic motif (RERRRKK or RERRGKK).Conclusion One of the reasons for H5N1 antibody positive rate of 25.5％ among poultry markets workers was that there were large numbers of H5N 1 subtype AIV detected in the environment of poultry markets and HA genes of H5N 1 subtype AIV in the poultry markets environment carried molecular characteristics of highly pathogenic which could increase the risk for H5N1 subtype AIV transmission in the environment of poultry markets.

BACKGROUNDWide application of umbilical cord blood transplantation (UCBT) in adult patients is limited by low cell-dose available in one umbilical cord blood (UCB) unit. The aim of this study was to investigate the safety and long-term outcomes of UCBT from unrelated donors in adult and adolescent patients with leukemia.

METHODSThirteen patients with leukemia received double-unit UCBT with human leukocyte antigen (HLA) mismatched at 0 - 2 loci. We analyzed the engraftment, graft-versus-host disease (GVHD) and survival.

RESULTSTwelve evaluable patients (92.3%) had neutrophil and platelet engraftment at a median of 21 days (range, 16-38 days) and 34 days (range, 25 - 51 days), respectively. At day 30, engraftment was derived from one donor in 8 patients (66.7%, 95%CI 40.0% - 93.4%), and from both donors in 4 patients (33.3%, 95%CI 6.7% - 60.0%) with 1 unit predominated. Unit with larger nucleated cell (NC) dose would predominate in engraftment (P = 0.039), whereas CD34(+) cell dose or HLA-match failed to demonstrate any relationship with unit predominance. Only one patient developed grade II acute graft-versus-host disease (aGVHD). Chronic GVHD (cGVHD) was observed in 2 of 11 patients who survived more than 100 days, and both were limited. The median follow-up after transplantation for the 13 patients was 45 months (range 1.5 - 121.0 months) and 72 months (range 41.0 - 121.0 months) for the 8 alive and with full donor chimerism. The 5-year cumulative disease free survival (DFS) was (61.5 ± 13.5)%. Of the 13 patients, 5 patients died in 1 year and 1-year transplantation related mortality (TRM) was 23.1% (95%CI 0.2% - 46.0%).

CONCLUSIONDouble-unit UCBT from unrelated donors with HLA-mismatched at 0-2 loci may overcome the cell-dose barrier and be feasible for adults and adolescents with leukemia.

Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; adverse effects ; methods ; Disease-Free Survival ; Female ; Graft vs Host Disease ; etiology ; Humans ; Leukemia ; immunology ; mortality ; therapy ; Male ; Treatment Outcome ; Young Adult

The aim of this study was to investigate the incidence, risk factors of acute renal failure (ARF) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), and evaluate its effect on the prognosis of patients after allo-HSCT. A retrospective analysis was performed in 86 patients undergoing allo-HSCT at Peking University First Hospital from June 2003 to April 2007. ARF is defined as a doubling of baseline serum creatinine at any time during the first 100 days post-transplant. The risks of ARF and mortality after ARF were examined using univariate analysis and multivariate unconditional logistic regression. The correlation of ARF and survival was examined using Cox regression. The results indicated that 27 patients (31.40%) developed ARF at a median of 59.5 days after transplant (range 1 to 93 days). The univariate analysis showed that elevated risks were severe acute GVHD (OR 6.196; 95% CI 1.121 - 34.249, p = 0.033), sepsis or septic shock (OR 4.184; 95% CI 1.314 - 13.325, p = 0.018) and hyperbilirubinemia (OR 3.709; 95% CI 1.428 - 9.635, p = 0.006). Renal disease before transplant (OR 6.711; 95% CI 1.199 - 37.564, p = 0.027), hypertension (OR 2.067; 95% CI 0.739 - 5.782, p = 0.165), the use of vancomycin (OR 2.133; 95% CI 0.844 - 5.392, p = 0.106) or foscarnet sodium (OR 2.133; 95% CI 0.844 - 5.392, p = 0.106) may be potential risks. Multivariate logistic regression analysis showed that renal disease before transplant (OR 6.288; 95% CI 1.218 - 32.455, p = 0.028), sepsis or septic shock (OR 3.614; 95% CI 1.040 - 12.544, p = 0.043) and hyperbilirubinemia (OR 4.448; 95% CI 1.563 - 12.665, p = 0.005) appear to be independently associated with an increased risk of ARF. Age, gender, baseline serum creatinine level, advanced malignant disease, unrelated-donor, total body irradiation (TBI) and cyclosporine levels were not associated with the development of ARF. Cox regression showed that ARF (RR 2.124; 95% CI 1.016 - 4.441, p = 0.045) was independently associated with survival of patients after allo-HSCT. The mortality of patients with ARF within 6 months post-transplant was significantly higher than that of those without ARF (44.4% vs 8.47%, p < 0.001). It is concluded that the cumulative incidence of ARF after allo-HSCT remains high. Renal disease before transplant, hyperbilirubinemia and sepsis or septic shock are all related factors which can increase the risk of ARF. ARF appears to be independent factor influencing survival of patients after allo-HSCT.
Acute Kidney Injury ; etiology ; Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Incidence ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Transplantation, Homologous ; Young Adult

In order to investigate the transmission risk of H5N1 avian influenza viruses (AIV) from sewage in Changsha poultry markets, the evolution relationship and molecular characteristics of non-structural (NS) genes of H5N1 AIV from sewage were analyzed. Nine H5N1 AIV environmental sewage specimens were collected from Changsha poultry markets. The NS genes were amplifyed by PCR and then sequenced with TA cloning. Amino acid(aa) sequence alignment and phylogenetic tree analysis were conducted by Lasergene and Mega5 software. Eight NS genes TA cloning were constructed successfully. Phylogenetic tree indicated that they were belonged to the allele A subgroup. Aa homology analysis showed 90.1% 92.5% identity in NS1 proteins and 91.0% - 92.6% identity in NS2 proteins compared with reference viruses of the allele A (A/chicken/ Hubei/ w h/ 1999). The homologies of the amino sequences of NS1 and NS2 in this study were 93.8%-100.0% and 98.4%-100.0%, respectively. The C terminal of all eight H5N1 NS1 proteins from sewage in poultry markets carried a ESEV of PL motif and the 92 amino acids were E, furthermore, the 80 to 84aa were missed which were the characteristics of highly pathogenic AIV. The NS genes of H5N1 AIV from sewage in poultry markets have molecular characteristics of highly pathogenic and have the potential risk of H5N1 virus spreading.
Amino Acid Sequence ; Animals ; Influenza A Virus, H5N1 Subtype ; chemistry ; classification ; genetics ; isolation & purification ; Influenza in Birds ; transmission ; virology ; Molecular Sequence Data ; Phylogeny ; Poultry ; Sequence Homology, Amino Acid ; Sewage ; virology ; Viral Nonstructural Proteins ; chemistry ; genetics

OBJECTIVETo observe the engraftment, survival and graft-versus-host disease (GVHD) after 2 units unrelated cord blood (UCB) transplantation for treatment of adult hematological malignancies.

METHODSAmong twelve patients with hematological malignancies, ten were in stable stage and 2 in advanced stage. Conditioning regimen was Bu/Cy or Cy/TBI in 11 cases, and 1 received nonmyeloablative regimen. Antithymocyte globulin (ATG) was administered in all patients. GVHD prophylaxis consisted of cyclosporine A (CsA), mycophenolate mofetil (MMF) and short course methotrexate (MTX). Each patient received 2 units UCB of HLA mismatched at 0 -2 loci. Median total nucleated cells (TNC) infused was 5.55 x 10(7)/kg [range (2.99 -8.18) x 10(7)/kg].

RESULTSOne patient showed primary graft failure. The other 11 patients showed neutrophil engraftment at a mean time of (21.6 +/- 5.1) days and platelet engraftment at (34.9 +/- 9.5) days. One patient showed secondary graft failure and died of leukemia relapse at 3 months after transplantation. Ten patients (83.3%) gained sustained engraftment. In 9 patients the UBC unit with larger TNC dose predominated engraftment, and only 1 patient showed the unit with smaller TNC predominated (P = 0.011). Acute GVHD was observed in 6 patients, including grade I in 5 and grade II in 1. Limited chronic GVHD was observed in 2 of 10 patients survived more than 100 days. Of the total 12 patients, eight were still alive in event-free status and 3-year event-free survival(EFS) was (66.7 +/- 13.6)%. Of the 10 patients in stable stage at the time of transplantation, the probability of EFS was (70.0 +/- 14.5 )%.

CONCLUSIONSTwo UBC units transplantation with HLA mismatched at 0 - 2 loci is feasible as a treatment modality for adult hematological malignancies, and the unit with larger TNC dose would predominate the engraftment.

Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; Female ; Follow-Up Studies ; Graft vs Host Disease ; prevention & control ; Hematologic Neoplasms ; drug therapy ; therapy ; Humans ; Male ; Survival Rate ; Transplantation Conditioning ; Young Adult