Objective To study a rapid and sensitive method for determination of virulence-associated genes in O1El Tor,O139,non-O1/non-O139Vibrio cholerae strains.Methods Five pairs of primers were designed respectively ac-cording to cholera toxin sub-unit A gene(ctxA),accessory cholera enterotoxin gene(ace),zonula occludens toxin gene(zot),toxin coregulated pilus A gene(tcpA)and toxR regulatory protein gene(toxR).Multiplex PCR(MPCR)procedures for simultaneously detecting those five genes were established.The gene information of the virulence-associated genes in the Vibrio cholerae strains was obtained through agar gel electrophoresis for products of single amplification of the MPCR.Results The five virulence-associated genes in the positive control Vibrio cholerae O139(MO45strain)could be detected and the results were correct,which could meet the designed request for the method.In the other tested strains(O1EL Tor,O139,non-O1/non-O139)could be detected1to5kinds of the virulence-associated genes.Based on the results of the variety of carried virulence-associated genes,the tested Vibrio cholerae strains could be classified as5kinds of genetypes,and the Vibrio cholerae could be distinguished between toxic and non-toxic strains.The sensitivity of the MPCR approach reached to10 2 cfu/ml.Conclusion This method is rapid,specific and sensitive,which possess great value for practical application.

Objective To explore whether there is a visual spatial attention span deficit in children with Chi-nese language learning disability and the characteristics of brain electrophysiological dynamic change. Methods Adop-ting event-related potentials ( ERP ) technology and levels of visual spatial attention span with fixed location cuing pa-radigm were used to explore the characteristics of brain electrophysiological dynamic change. Results Based on the behavioral data,the repeated findings of analysis of variance on reaction time showed that both the main effect of the type of subjects[F(1,35)=4. 34,P<0. 05,η2=0. 11],and the main effect of attention span[F(1,35)=138. 75,P<0. 001,η2=0. 79] had statistical significance. The repeated analysis of variance on accuracy showed that the main effect of subjects type[F(1,35)= 4. 99,P<0. 05,η2=0. 13],and the main effect of attention span[F(1,35)=101. 83,P<0. 01,η2=0. 74] were significantly different. Under large and small attention span conditions,the children with Chinese language learning disability showed a longer time of reaction and a lower accuracy rate than the children in the control group. With the electroencephalograph( EEG) data and under large and small attention span conditions,the Chinese learning disability children showed enhanced P1 amplitude and reduced N1 amplitude. Conclusions There is a visual spatial attention span deficit in children with Chinese learning disability which may be caused by a weak prima-ry sensory processing ability and divided attention deficit.

Objective To promote rational drug use and assure the drug use in an effective and safe way by developing the monitoring role of clinical pharmacists in the adverse drug reaction .Methods The common reasons for adverse drug reaction were an-alyzed.The clinical pharmacists took advantage of their professional knowledge to deal with adverse drug reactions and reinforce the monitoring of the drug treatment.Results The clinical drug safety was ensured by reducing the adverse drug reaction incidence rate. Conclusion Pharmacy information exchange should be enhanced and the professional knowledge of clinical medical staff should be broadened to promote clinical rational drug use.

Objective To explore quality of life (QOL) of spouses of spinal cord injury patients,as well as its influencing factors.Methods Sixty-eight spouses of spinal cord injury patients were recruited as an observation group.At the same time,another 70 healthy married people who had no serious chronic diseases in the family but were matched in gender,age,occupation and education were chosen as a control group.The QOL of all the subjects in both groups was investigated by way of a questionnaire survey.The results of both groups were compared and factors that influenced their QOL were explored.Results The QOL of spouses of spinal cord injury patients,including total QOL ( 3.22 ± 0.37 ),physiological function ( 60.57 ± 14.22 ),psychological state ( 60.09 ± 8.06 ),social relations (63.37 ± 8.10 ) and the living environment (66.86 ± 11.26 ) were significantly lower than in the control group (4.62 ± 0.75 ; 82.48 ± 10.92 ; 86.02 ± 8.75 ; 85.94 ± 10.13 and 83.23 ± 9.54,respectively).The QOL of spouses of spinal cord injury patients was closely related to the patient's injury type,injury position,the treatment protocol,as well as their own gender,vocation and education.Conclusion The QOL of spouses of spinal cord injury patients is significantly lower than that of normal people,and closely related with the patients' injury type,injury position and treatment,as well as their own gender,vocation and education.

Objective To investigate the diagnostic value of T-SPOT.TB in patients with fever.Methods T-SPOT.TB was carried out in 456 febrile patients who had no determined reason for fever or who could not be excluded from infection with Mycobacterium tuberculosis.These patients were followed up for their final clinical diagnosis and the diagnostic value of T-SPOT.TB was evaluated using the receiver operating characteristic (ROC) curve.Results In the present study,85 patients were clinically diagnosed with tuberculosis (TB) (including 34 cases of pulmonary TB,24 cases of TB infection defined as T-SPOT.TB positive without TB lesion,but effective with anti-TB treatment,10 cases of tuberculous pleurisy and 17 cases of other extrapulmonary TB) and 371 patients had no definite evidence of Mycobacterium tuberculosis infection (including 42 cases of suspected TB and 329 cases of other diseases).T-SPOT.TB had a high sensitivity of 84.3％ in TB patients and a high negative predictive value of 94.2 ％,but the specificity was 68.1％ and the positive predictive value was 41.4 ％.The Kappa value was 0.380 and the area under ROC curve was 0.762 (95％CI:0.707-0.817).T-SPOT.TB had a higher sensitivity in extrapulmonary TB than in pulmonary TB (88.2 ％ vs 78.1 ％),with the negative predictive values of 95.3 ％ and 92.6 ％,Kappa values of 0.438 and 0.299,and the areas under ROC of 0.791 (95 ％ CI:0.724-0.857) and 0.719 (95％CI:0.623-0.815),respectively.Ninety-nine cases were T-SPOT.TB positive out of 329 patients diagnosed with other diseases.Conclusions T-SPOT.TB test can be used as a method for TB diagnosis in patients without obvious TB lesions.Due to its relatively high negative predictive value,T-SPOT.TB can be used to rule out infection with Mycobacterium tuberculosis.

Objective To study cellular immune responses induced by DNA vaccine against Chlamydia trachomatis (Ct) serotype E. Methods BALB/c mice were divided into three groups to be intramuscularly immunized by blank plasmid (negative control group), DNA vaccine against Ct serotype E (vaccine group), and inactivated Ct elementary body (positive control group), respectively. Two weeks after the last immunization,delayed-type hypersensitivity (DTH) response was evaluated; MTT assay was performed to detect the proliferation of spleen lymphocytes, ELISA to measure the serum level of interferon-γin mice. Some immunized mice underwent a genital challenge with Ct elementary body followed by isolation of Ct from exfoliated epithelial cells in genital tract and pathological examination of cervical tissue from the challenged mice. Results Compared to negative control group, vaccine group and positive control group experienced a stronger DTH response.The lymphocyte stimulating index and serum level of IFN-γwere highest in the positive control group (3.81 ±0.30, 2891.7 ± 1048.8 μg/L), followed by vaccine group (2.35 ± 0.25, 593.3 ± 342.6 μg/L) and negative control group (1.48 ± 0.15, 309.2 ± 157.9 μg/L), and significant difference was observed between the three groups (P ＜ 0.05 or 0.01 ). After Ct challenge, Ct was isolated from exfoliated epithelial cells and cervical tissue was damaged in the negative control group, while in the other two groups, Ct was undetected and genital tract tissue was intact. Conclusions The DNA vaccine against Ct serotype E could induce Ct-specific cellular immune responses to some extent, and offer a protection against vaginal challenge with Ct.

Objective: To investigate six mineral element contents of rice bran and milled rice from three color rice cultivars (white rice, red rice, black rice). Methods: Select 36 typical cultivars and determine Se, Zn, Fe, Ca, Mn, Cu contents in their rice bran and milled rice with AFS-2201 atomic fluorescent spectrometer and AA-680 atomic absorption spectrometer. Results: 1. The six mineral element contents of rice bran were 2.58, 3.35, 10.04, 4.58, 12.79 and 6.59 times as those of milled rice respectively. 2. The Ca content of black milled rice was significantly higher than that of white and red milled rice. The Mn content in rice bran of white rice was significantly higher than those of black and red rice. Other mineral element contents were not significantly different. 3. For rice bran and milled rice of different cultivars, the contents of each mineral element in three color rices varied greatly. Conclusions: 1. The content of six minerals in rice bran was 5.66 times on average as that in milled rice in all cultivars of three color rice. 2. Eight cultivars of the highest content of each mineral in rice bran and milled rice were recommended for commercial use and human consumption.

Background Dry eye is a common disease worldwide.Cyclosporine A(CsA) is provided to be a immunosuppressive agent and is effective on dry eye.But in China,0.05％ CsA is not yet applied in dry eye treatment.Objective This study was to evaluate the efficacy and safety of 0.05％ CsA eye drops in the treatment of dry eye.Methods This was a randomized,double-blind,vehicle-controlled parallel group study.Forty eyes of 40 patients with moderate to severe dry eye were randomly divided into two groups,with the corresponding treatment of 0.05％ CsA eye drops or the vehicle emulsion.The patients in both the groups received non-preserved artificial tear.Symptoms and signs were observed before administration,(7±1),(28±2),(56±3),and (84±3) days and also 14 days after withdrawal.The clinical effective rate was considered as the primary outcome.The subjective assessment of the patients including total symptom scores and ocular surface disease index (OSDI) scores,Schirmer Ⅰ test (S Ⅰ t) with topical anaesthesia,tear film breakup time (BUT),rose Bengal and fluorescein staining scores were evaluated.The safety profile was evaluated by adverse events,visual acuity and ocular tolerance.Results At the end of this trial,the ocular symptoms scores,conjunctival hyperemia,BUT,S Ⅰ t and keratoconjunctiva staining scores of the two groups had statistically significant difference.The total effective rate of 0.05％ CsA treatment group was 75％ (15/20) and vehicle group was 25％ (5/20).There was a statistically significant difference between groups (P =0.000),and the 95％ confidence interval (C1) of the difference value of total effectiveness between the two groups was 30.80％-53.75％.At the end of this trial,there was no statistically significant difference in visual acuity distribution (P =0.890).No obvious discomfort was found in the patients received 0.05％ CsA eye drops.There were no adverse events during the follow-up duration.Conclusions 0.05％ CsA ophthalmic emulsion is an effective and safe treatment for dry eyes.

Objective Establish premature ovarian failure ( POF ) model in female Sprague Dawley by tripterygium wilfordii , and investigate the effect of bushenyangxue prescription on the levels of anti -mullerian hormone ( AMH) .Methods After POF model was established , rats were gave by gavage of different dosage of Bushenyangxue prescription for 30 days.The changes of histomorphology on rat ovarian tissue were observed by hematoxylin -eosin staining. Serum AMH concentraction , protein and mRNA expression of AMH were measure with ELISA , immunohistochemical staining and qRT-PCR, respectively .Results The follicle and corpus luteum were atrophied after tripterygium wilfordii challenge, which was improved after treatment with Bushenyangxue prescription .Serum AMH, protein and mRNA expression of AMH were decreased tripterygium wilfordii-treated rats; this decrease was inhibited after treatment with Bushenyangxue prescription .Conclusions Our study indicates that Bushenyangxue prescription could preserve the AMH levels of POF rats.These findings suggest that Bushenyangxue prescription may be a useful strategy to treat POF .

ObjectiveTo study the effect of a recombinant plasmid encoding mouse interleukin 2 (mlL-2) on the immunogenicity of DNA vaccine against Chlamydia trachomatis(Ct) serovar E.Methods BALB/c mice were divided into 4 groups to be intramuscularly inoculated with blank plasmid(negative control group),DNA vaccine against Ct serovar E(DNA vaccine group),DNA vaccine against Ct serovar E and a recombinant plasmid containing mIL-2(combination group),and inactivated Ct serovar E elementary bodies (positive control group),respectively.The immunological effects were evaluated by posterior foot pad thickness,proliferation level of spleen lymphocytes,serum level of IL-4 and interferon (IFN)-γ in mice,and the capability to clear Ct genital tract infection.ResultsThe proliferation index of spleen lymphocytes in the combination group and positive control group was similar(3.64 ± 0.41 vs.3.77 ± 0.34),but was significantly different from that in the blank control group and DNA vaccine group (1.37 ± 0.21 and 2.52 ± 0.30).The serum level of IL-4 was(38.49 ± 12.24) pg/ml in the positive control group,significantly higher than in the negative control group,DNA vaccine group and combination group ((25.37 ± 18.93),(24.75 ± 8.49),(21.74 ± 6.43) pg/ml,respectively).With respect to the serum level of IFN-γ,the combination group and positive control group were similar ((1923.3 ± 518.1) pg/ml vs.(2712.5 ± 887.2) pg/ml),but were significantly different from the negative control group and vaccine group((310.8 ± 160.7) pg/ml and(601.3 ± 357.9) pg/ml).Six days after Ct challenge,the exfoliated cells from genital tract were positive for Ct culture in the negative control group,but negative in the other 3 groups.ConclusionIL-2 genetic adjuvant can enhance the immune response,especially Th1 type response,induced by the DNA vaccine against Ct serovar E.