Objective:To construct a recombination plasmid containing a kanamycin resistance gene,the upstream and downstream fragment of luxS of Streptococcus mutans so that luxS can be knock out by transforming the plasmid into S.mutans later.Methods:Kanamycin resistance gene,the upstream and downstream of luxS were cloned respectively by using plasmid pEGFP-N1 and DNA of Streptococcus mutans as template.Then the genes were ligated into Multiple Cloning Site(MCS) of vector pMD19-T in certain order and transformed into E.coli Competent Cells.Finally transformants were selected for resistance to kanamycin and ampicillin.Results:Kanamycin resistance gene and the upstream and downstream of luxS were successfully ligated into accurate enzyme digestion site of vector pMD19-T,and restriction digests analysis and sequencing result was correct.Conclusion:LuxS gene knock-out of Streptococcus Mutans recombinant plasmid is constructed and built a base of constructing Streptococcus Mutans luxS mutans in the future.

Objective To observe the clinical efficacy of acupoint application in canicular days plus TDP radiation in treating primary dysmenorrhea due to cold-dampness.Method Eighty-three patients were randomized into group A of 32 cases, group B of 20 cases, and group C of 31 cases. Group A was intervened by acupoint application plus TDP, group B was by acupoint application alone, while group C was by oral administration ofTong Jing Bao granules. The Visual Analogue Scale (VAS) scores of the three groups were observed before and after treatment, and the clinical efficacies were compared.Result The total effective rate was 93.1% in group A, versus 85.0% in group B and 77.4% in group C. The total effective rate of group A was significantly different from that of group B and group C (P<0.05). The total effective rate of group B was significantly different from that of group C (P<0.05). The VAS scores were changed significantly after treatment in the three groups (P<0.05). After treatment, the VAS scores of group A and B were both significantly different from that of group C (P<0.05).Conclusion Acupoint application plus TDP is an effective method in treating primary dysmenorrhea due to cold-dampness.

In this paper, some items about the microbes listed in the current National Standard of Laboratory Animals were reviewed, including their host spectrum, impact of infection on the animals, their interference on research works and their epidemiology in laboratory animals.This paper may provide some clues for the update of our National Standard of Laboratory Animals.

Objective To evaluate the clinical efficacy and safety of a fractional bipolar radiofrequency device for the treatment of wrinkles of the face and neck.Methods This study enrolled 39 volunteers (including 37 females and 2 males aged 35-60 years) with Fitzpatrick skin type Ⅳ-Ⅴ and Fitzpatrick wrinkle scale score of 4-6.All the subjects received three sessions of treatment with a fractional bipolar radiofrequency device at intervals of 4-6 weeks.Follow-up visits were scheduled at one month after each treatment session for evaluation of efficacy (using standardized photography),subjective satisfaction and adverse effects.Results After three sessions of treatment,all the subjects experienced a significant decrease in Fitzpatrick wrinkle scale compared with baseline values,and wrinkle improvement score increased with the increase in treatment sessions.The Fitzpatrick wrinkle scale score for forehead wrinkles,glabella wrinkles,fishtail lines and neck wrinkles in these subjects after three sessions of treatment significandy differed from that before treatment (all P ＜ 0.01).Significant differences were also observed in the wrinkle improvement score for forehead wrinkles between these subjects after two sessions and one session of treatment (P ＜ 0.01),and observed in that for glabella wrinkles,fishtail lines and neck wrinkles between these subjects after three sessions and one session of treatment (P ＜ 0.05 or 0.01).Side effects were mild with no significant downtime.Conclusion Fractional bipolar radiofrequency may be a safe and effective option for the treatment of wrinkles of the face and neck in individuals with Fitzpatrick skin type Ⅳ-Ⅴ.

Objective To check the maneuverability of the nursing safety management indicators for skilled nursing facilities.Methods A hospital based skilled nursing facility in Chongqing was chosen as the main study target while a Red Cross nursing hospital in Shanghai which operated earlier was also included as a comparison object.Combination of files reference,on-the-scene observation,manager interviews,sampling survey and institution's self-assessment,scores of indicators were calculated,and the total scores for both institutions were calculated as well.Results The institution from Chongqing marked 81 points while the other one got 86 points.Conclusions The indicator system had good maneuverability and the weight of indexes was consistent with the practical requirement of safety management.This indicator system could provide reference for standardize skilled nursing facilities' management,but still needed further revision and consummation.

Objective To explore the clinical features and gene mutations of blepharophimosis-ptosis-intellectual-disability syndrome (BPID). Methods The clinical data, diagnosis and treatment of a child with BPID in neonatal intensive care unit (NICU) were reviewed. Based on the literature retrieved from PubMed database, the common classification, clinical features, diagnosis and genetic counseling of BPID and its affiliated blepharophimosis-mental retardation syndromes (BMR) were reviewed. Results This male infant was 39 weeks of gestational age with birth weight of 1920 g, and was admitted to NICU 15 min after birth due to dyspnea. The main clinical manifestations were facial deformity such as biepharophimosis, ptosis and micromandible, inspiratory dyspnea with laryngeal cartilage softening, malformations of the thorax and feeding difficulties. A heterozygous mutation in UBE3B gene was identified by complete exon sequencing and he was diagnosed of BPID, a rare genetic disorder. Reviewing the literature, there was no relevant report in domestic. While one foreign literature was found to report 5 patients from 4 families having a subtype of BMR, a kind of autosomal recessive diseases caused by mutations in the UBE3B gene. Conclusion BPID is a rare clinical entity of BMR. Complete exon sequencing can be used to diagnose the disease.

Objective To study the effects of the intensive treatment with continuously subcutaneous in-sulin infusion(CSII) in patients with gestational diabetes and the nursing during the treatment. Methods From Jan 2007 to Aug 2008, 42 patients with gestational diabetes underwent continuously subcutaneous insulin infu-sion, and a standard nursing plan of patients was made up before treatment. The efficacy of treatment and nursing were analyzed. Results After using the CSII, the blood glucose of all patients was controlled at normal level after 7.0±1.5d. 88.1% of patients were satisfied with the effect and one patient was dissatisfied with the treatment because of losing the fetus for sever bleeding caused by placenta previa. 40.5% of patients with different degrees of anxiety accepted CSII therapy after psychological care by nurses;In 9.5% of patients the infusion bump a-larmed because of needle blockage or infusion tube bending, it restarted working and ran normally once the needle replaced or bending tube unwinded. 7.1% of patients developed hypoglycemia during CSII and recovered after eating. Conclusions Intensive treatment with CSII in gestational diabetes patients can effectively control blood glucose level The effect can be improved by taking specific nursing measures.

Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .

Objective:To investigate the activation of peripheral basophils from patients with rheumatoid arthritis ( RA) and its mechanisms. Methods:The activation markers including CD203c and the proportion of IL-4 positive rate of peripheral basophils from RA patients and healthy controls were detected by flow cytometry. Serum levels of IgE in RA patients and healthy controls were detected by electrochemilu minescence immunoassay. Basophils were negatively isolated and co-cultured with or without purified IgE,anti-IgE as positive control,then,the expression of CD203c and propotion of IL-4 positive basophils were detected by flow cytometry. Results:The expression of CD203c and IL-4 positive rate of basophils from RA patients were higher than that of healthy controls (P<0. 05). Serum levels of IgE in RA were higher than that of healthy controls (P<0. 05). After co-cultured with isolated IgE from RA patients,basophils negatively isolated from healthy controls were activated,and higher expression of CD203c and proportion of IL-4 (P<0.05). Conclusion:Basophil activation is related with development of RA,and its activation is mainly mediated by IgE. Targeting basophil and its activation pathway would be expected to provide new strategies for the treatment of RA.

Objective To analyze the current situation and trends of nursing home research to provide reference for further study.Methods The GoPubMed retrieval tool was used to retrieve the research literatures on the nursing home in PubMed database with the subject term "Nursing Homes",then the Excel2010 was used to statistically analyze the items such as the published literature's time distribution,geographical distribution,journals distribution,high-authors,related subjects.Results In total,46 436 articles were retrieved out.The publication amount was increased year by year.The largest publication amount was the United States,but China ranked at 21.Meanwhile the top 10 cities,top 10 journals and top 10 authors in publication amount were mainly in European and American countries,while China did not enter the ranks.The main research fields were community care,long-term care,etc.Conclusion In recent years,the research literatures about nursing homes present upward trend as a whole,but which are mainly concentrated in European and American countries.China is relatively weaker,so there is still much rising space.