Objective To improve the sensitivity of PCR for studying the rifampin resistant gene (rpoB) of M.Leprae from clinical samples.Method After comparing the results of PCR with Q-Solution (PCR Enhancer) and without Q-solution,Nested-PCR was then applied for the detection of rpoB gene.Results Although PCR Enhancer improves the amplification of rpoB gene, the sensitivity of routine PCR is only 45. 2%.The sensitivity of Nested-PCR for detecting mutants in rpoB gene can further increase to 90.5% after selection of optimum parameters.Conclusion The combinaion of PCR Enhancer and Nested-PCR improves the sensitivity and specificity of PCR for detection of rifampin resistant gene of M. Leprae.

AIM: To study the pathological change in mouse organs immunitied by inactivated SARS-CoV vaccine. METHODS: Inactivated SARS-CoV vaccine was injected into BALB/c and C57BL/6 mice. Anti-SARS antibody was analyzed by ELISA. After 8 weeks, the immunitied mice were killed and those organs were analyzed by pathological methods. RESULTS: Anti-SARS antibody in mice was positive after 8 days. Only minimal injury was observed in a few lungs and livers, but the other organs were not. CONCLUSIONS: Inactivated SARS-CoV vaccine induced mice to create antibody, whereas they did not cause severe injury. This result will be valuable for vaccine into clinical research. [

Objective To study the relationship between bronchial asthma and smoking status in Chinese people.Methods Asthma epidemiological survey and stratified-cluster-random method survey were performed in residents over 14 years in 8 provinces (cities) of China from February 2010 to August 2012.Asthma was diagnosed based upon case history,clinical signs and lung function test.Smoking status was investigated by questionnaire.Results Sampling population was 180 099 and 164 215 were valid.A total of 2 034 subjects were diagnosed as asthma including 79 692 men and 84 523 women.The overall prevalence rate of asthma was 1.24％ (2 034/164 215).Smokers were 23.8％ (39 137/164 215) in the whole population.Smokers were 34.5％ (702/2 034) in asthmatic patients,compared with 23.7％ (38 435/ 162 181) in no-asthmatic population.The incidence of asthma was 1.79％ and 1.06％ in smokers and nonsmokers respectively (P ＜0.001),suggesting that OR of smoking was 1.70 (95％ CI 1.55-1.86,P ＜ 0.001).According to asthma control test (ACT) score,the level of asthma control in non smoking group was higher than that in smoking group(43.2％ vs 35.3％).The times of hospitalization due to acute exacerbations (0.51 vs 0.41 events/person/year),total hospitalization rate (27.35 ％ vs 20.12％),annual emergency room visits (0.80 vs 0.60 events/person/year) and emergency room visit rate (31.77％ vs 24.47％) were all much higher in smoking asthmatic patients than those in non smoking asthmatic patients,indicating that the level of asthma control in smoking patients was significantly worse than in non smoking patients.Conclusions The smoking rate in Chinese people over 14 years is still high.The prevalence rate of asthma in smokers is significantly higher than that of non-smokers.The level of asthma control in smokers is significantly worse than that in non smokers.

OBJECTIVETo evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases.

METHODSVero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells.

RESULTSFive pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect.

CONCLUSIONsiRNA may be effective in inhibiting SARS-associated coronavirus replication.

Animals ; Cercopithecus aethiops ; RNA, Small Interfering ; pharmacology ; SARS Virus ; drug effects ; Transfection ; Vero Cells ; Virus Replication ; drug effects

Objective To provide scientific evidence to identify and confirm severe acute respiratory syndrome (SARS) by laboratory detection.Methods Multiple clinical specimens were collected serially and systematically from the 4 suspected SARS patients, which occurred between Dec.2003 to Jan.2004 in Guangdong Province. The samples were tested by serologic and molecular methods.Results IgM or IgG antibodies against SARS-CoV were detectable after 6—8 days of the onset in four patients. The four-fold or greater rising in antibodies was clearly detected in three of the four patients, while the fourth patient’s seroconversion was from negative to positive. The results analysed by enzyme-linked immunosorbent assay( ELISA), immunoflourescence assay (IFA), and neutralization test were highly correlated. SARS-CoV RNA was just detected in 3 throat swab specimens from case 1 by real-time PCR. M, N and S genes were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) from the positive samples. Sequencing results showed that they were SARS-CoV gene segments, and most closely matched SARS-CoV gene sequences were isolated from civet cats in Guangdong Province. Nevertheless, SARS-CoV was not isolated from any samples of the 4 patients.Conclusion Based on these results, the 4 reported cases were laboratorily confirmed as SARS cases.

Objective : To explore from the perspective of microorganisms the changes in plaque microbial community of children with severe early childhood caries (S-ECC) before and 3 months after dental treatment. Meanwhile to show the effect of treatment on the maintenance of long- term caries-free state. Methods: S-ECC children completed dental treatment under general anesthesia. We collected plaque from caries-free dental surfaces before treatment (caries, C) and at the postoperative follow-up review time points of 7 days (C-7D), 1 month (C-1 M), and 3 months (C-3 M). We included caries-free children (caries free, CF) as the control group to analyze the dynamic modification process of the plaque microbial community in the short-term pre- and postdental treatment. Results: Species clustering analysis showed that the compositions of the microbial communities of the S-ECC and CF groups were highly similar. The α diversity index was not statistically significant (P>0.05). From the analysis of the relative abundance, Leptotrichia spp. and Aggregatibacter spp. decreased after treatment compared with before treatment (P<0.05). Streptococcus sanguinis in the C-7D group increased compared with that in the C group and gradually decreased within 3 months. Veillonella spp., Actinomyces spp., Allprevotella spp., Capnocytophaga spp., and Streptococcus mutans differed between the C and CF groups (P<0.05), Streptococcus mutans did not differ significantly between the C-7D and C-1 M groups and the CF group after treatment, while C-3 M showed an increase compared with the CF group (P<0.01). Conclusion The rapid change in the structure of the flora of children with S-ECC after treatment. The plaque microbial community structure in a caries-free state gradually starts to be established 1-3 months after treatment. There is a "core microbiota" in the oral plaque community that jointly maintains microecological stability. Veillonella spp., Allprevotella spp. and Streptococcus mutans have potential as possible microbial markers.

OBJECTIVE: To investigate the role of long-chain non-coding RNA MALAT1 in modulating paclitaxel resistance in breast cancer cells. METHODS: Breast cancer SK-BR-3 cells were treated with gradient concentrations of paclitaxel to induce paclitaxel resistance of the cells. The resistant cells were transfected with si-NC, si-MALAT1, pcDNA, pcDNA-MALAT1, miRNC, miR-485-3p mimics, si-MALAT1+anti-miR-NC, or si-MALAT1+anti-miR-485-3p liposomes. Following the transfections, the cells were examined for changes in IC of paclitaxel using MTT assay; the protein expression of P-gp, Bcl-2 and Bax were detected with Western blotting, and a dual luciferase reporter assay was used to detect the binding of MALAT1 to miR-485-3p. RESULTS: Compared with paclitaxel-sensitive SK-BR-3 cells, paclitaxel-resistant SK-BR-3 cells showed significantly increased the IC of paclitaxel with up-regulated MALAT1 expression and down-regulated miR-485-3p expression ( < 0.05). Silencing MALAT1 or overexpressing miR-485-3p obviously lowered the IC of paclitaxel and the expression of P-gp and Bcl-2 and increased the expression of Bax in SK-BR-3/PR cells ( < 0.05). miR-485-3p was identified as the target of MALAT1, and inhibiting miR-485-3p significantly reverse the effect of MALAT1 silencing on IC of paclitaxel and the expressions of P-gp, Bcl-2 and Bax in SK-BR-3/PR cells ( < 0.05). CONCLUSIONS MALAT1 can modulate paclitaxel resistance in breast cancer cells possibly by targeting miR-485-3p to down-regulate P-gp and Bcl-2 and up-regulate Bax.
Cell Line, Tumor ; Humans ; MicroRNAs ; Paclitaxel ; RNA, Long Noncoding ; genetics

OBJECTIVETo evaluate relevant prognostic factors of unrelated single unit umbilical cord blood hematopoietic stem cell transplantation (sUCBT), and to explore the correlation between cryopreservation time of cord blood and cell viability and outcome of sUCBT.

METHODSRetrospective analysis of 137 patients undergoing sUCBT with cord blood provided by Shanghai Cord Blood Bank from Mar. 15, 2007 to Dec. 26, 2013 were performed in this study. The mean cryopreservation time of 137 units cord blood was 698(96-1968) days, with mean cell viability of 87.4% after thawing.

RESULTSNo statistical difference on cell viability, hematopoietic reconstitution, graft failure, acute graft versus host disease (GVHD) and overall survival (OS) was found between patients transfused with cord blood preserved below and above 2 years. The 5-year OS of patients transfused with cord blood preserved below and above 2 years were 55.6% and 67.9%, respectively (P=0.124). OS of the UCBT at 2011 and before, and after 2011 was 48.7% and 79.6%, respectively (P=0.001). Age above 16-year-old (RR=2.830, P=0.027) and UCBT at 2011 and before (RR=0.203, P<0.001) were two risk factors of treatment related mortality.

CONCLUSIONOutcome of sUCBT in China had significant improvement in recent 2 years. Cryopreservation time of cord blood had no statistical correlation to cell viability after thawing and clinical outcome.

Cell Survival ; China ; Cord Blood Stem Cell Transplantation ; Fetal Blood ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Retrospective Studies

OBJECTIVETo investigate the impact of cryopreservation duration of umbilical cord blood (UCB) on quality of hematopoietic stem cell and outcome of clinical transplantation.

METHODS605 units of UCB which had been used in clinical transplantation were previously cryopreserved for 820 (88-2651) days in average. UCB was detected for total nucleated cell count, CD34+ cells count, cell recovery rate, cell viability and CFU-GM after thawing.

RESULTSNo statistical correlation was found between cryopreservation duration and cell recovery rate, cell viability. CFU-GM decreased along with the extension of cryopreservation duration (P=0.011), ranging between 109.6 and 105.7/1 × 10⁵. There was no significant difference on hematopoietic reconstitution time, graft failure, acute GVHD and overall survival among groups with different cryopreservation duration.

CONCLUSIONCryopreservation duration has no significant effect on cell recovery rate, cell viability and clinical transplantation outcome. Extension of cryopreservation duration may reduce CFU-GM of stem cells with fluctaion still in normal range. UCB could maintain cell viability and function to achieve satisfactory clinical transplantation outcome even when thawed after 3 to 7 years' cryopreservation.

Cell Count ; Cell Survival ; Cryopreservation ; Fetal Blood ; Graft vs Host Disease ; Granulocyte-Macrophage Progenitor Cells ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Treatment Outcome