BACKGROUND:Relative to blood samples, mouth swab samples are more beneficial for apolipoprotein E gene polymorphism analysis among large cohorts. However, agreement has not yet been reached about how to extract genomic DNA form mouth swab samples.
OBJECTIVE:To develop an appropriate method to extract genomic DNA form mouth swab samples, which are suitable for apolipoprotein E gene polymorphism analysis.
METHODS:Fifty mouth swab samples from patients with sporadic Alzheimer’s disease were col ected. Magnetic nanoparticles and PicoDNA trace nucleic acid extraction kit were used to extract genomic DNA form mouth swab samples. And the purity and concentration of the genomic DNA extracted by the two methods were analyzed. Then PCR amplifications and DNA electrophoresis were performed to confirm whether the genomic DNA was able to amplify desired DNA fragments. DNA sequencing was applied to analyze apolipoprotein E gene polymorphisms.
RESULTS AND CONCLUSION:Genomic DNA extracted by the two methods was of high purity. The concentration of genomic DNA extracted by magnetic nanoparticles was higher than by PicoDNA trace nucleic acid extraction kit, and the difference had statistical significance (P<0.05). Al the genomic DNA were able to performed PCR amplifications to obtain desired PCR products, but results of DNA electrophoresis showed that DNA fragments were more clear by nanoparticles method. The results of DNA sequencing were the same by the two methods. The distribution ofε2,ε3,ε4 genotypes of apolipoprotein E gene was 6%, 71%, 23%, respectively. Magnetic nanoparticles were better than PicoDNA trace nucleic acid extraction kit for extracting genomic DNA form mouth swab samples for apolipoprotein E gene polymorphism analysis.

Objective To investigate the expression of aquaporins-3 (AQP3) in amniotic epithelial cells regulated by cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signal pathway and to explore the mechanisms of its expression.Methods The amniotic epithelial cells were collected from 30 patients who underwent elective caesarean sections at term with normal amniotic fluid volume and primarily cultured.The cultured cells were treated with (1) forskolin groups: different concentration (0,2.5,5,50 or 100 μmol/L) of forskolin treated cells for 2 hours,and the optimal concentration of forskolin treated cells with different time (0,1,2,10 or 20 hours) ; (2)SP-cAMP groups: different concentration (0,2.5,5,50 or 100 μmol/L) of SP-cAMP treated cells for 2 hours,and the optimal concentration of SP-cAMP treated cells with different time (0,1,2,10 or 20 hours); (3)H-89 groups: different concentration (0,5,10,50 or 100 μmol/L) of H-89 treated cells for 2 hours,and the optimal concentration of H-89 treated cells with different time (0,1,2,10 or 20 hours).The level of intracellular cAMP and activity of PKA were detected by using ELISA,and immunohistochemistry was used to detect the localization of AQP3,the protein expression of total cAMP-response element binding protein (CREB) and phospho-CREB (p-CREB) and AQP3 were assessed by western blot analysis.Cell proliferation was assessed by cell counting kit-8 (CCK-8)assay.Results (1) The brown staining of AQP3 was detected in both cell membrane and cytoplasm in each group.(2) There was no significant change of the cell proliferation rate among groups with different concentration of forskolin,SP-cAMP and H-89 treatment (P ＞ 0.05).(3) After different concentration of forskolin treated 2 hours,the expression of total CREB had no significant difference among them(P ＞ 0.05).While the expression of cAMP level,PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 2.5 μmol/L,5 μmol/L,50 μmol/L forskolin group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 5 μmol/L forskolin group were higher than that in 2.5 μmol/L and 50 μmol/L (P ＜ 0.05).The optimal forskolin concentration was 5 μmol/L.(4) After different concentration of SP-cAMP treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 5 μμmol/L,50 μmol/L SP-cAMP group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 50 μmol/L SP-cAMP group were higher than that in 5 μmol/L (P ＜0.05).The optimal SP-cAMP concentration was 50 μmol/L (5) After different concentration of H-89 treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were lower in 10 μmol/L,50 μmol/L and 100 μmol/L H-89 group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 10 μmol/L H-89 group were lower than that in 50 μmol/L,100 μmol/L (P ＜ 0.05).The optimal H-89 concentration was 10 μmol/L.(6) p-CREB and AQP3 protein expression were significantly lower in 5 μmol/L forskolin combined 10 μmol/L H-89 incubating 2 hours group when compared with 5 μmol/L forskolin,but higher than that in 10 μmol/L H-89 treated group (P ＜ 0.05).Total CREB was no significant difference among the three groups (P ＞ 0.05).Conclusion cAMP-PKA signal transduction pathway may regulate AQP3 protein expression in human amniotic epithelial cells.

Background Many methods are widely studied to improve the therapeutic effect of trabeculectomy for glaucoma,and the clinical effectiveness of the amniotic membrane application during trabeculectomy is one of the research hotspots.Objective This study was to evaluate the efficacy and safety between trabeculectomy with (TE-AMT) and without amniotic membrane transplantation (TE-noAMT) for treating glaucoma.Methods Articles published from 1965 to December 2010 were searched from The Cochrane Library,PubMed,EMBASE,CNKI,Chinese Biomedicine Database,internet by computer,and manual search then was performed according to the predetermined strategy.Randomized controlled trials (RCT) on TE-AMT and TE-noAMT were included.The quality of included articles were scored based on Jadad table,and the relative parameters and information were extracted,including author,design of the trials,country,number of eyes and patients,gender,follow-up duration,age,classification of glaucoma,baseline IOP and loss rate.The main analysis indicators were the percentage of IOP reduction,completely operative successful rate,qualified success rate and adverse events.The pooled estimates were carried out with RevMan version 5.0 software.Results Nineteen RCTs documents were reviewed by meta-analysis with the Jaded scores ≥3 in 2 papers and ＜3 in 17 papers.Total 977 eyes of 818 patients were included.The weighted mean differences (WMD) of the percentage of IOP from baseline were (WMD =8.47％,95％ CI:5.20-11.75) at 6 months and (WMD=9.37％,95％ CI:4.97-13.77) at 12 months postoperatively.Relative risk (RR) of complete success rate and qualified success rate at postoperative 6 months were (1.40,95％ CI:1.19-1.65) and (0.47,95％ CI:0.22-1.00),respectively.RR values of adverse event such as shallow anterior chamber,hyphema,hypotony and choroidal detachment,failed filtering blebs at 6,12 months were 0.51 (95 ％ CI:0.30-0.85),0.43 (95％ CI:0.20-0.92),0.51(95％CI:0.26-1.00) and 0.57(95％CI:0.14-2.31),0.31(95％CI:0.20-0.47),0.31 (95％ CI:0.17-0.55),respectively.Conclusions TE-AMT appears to have better efficacy in lowering IOP,increasing complete success rate and reducing adverse event I in comparison with TE-noAMT.

Objective To observe the efficacy of prophylactic use of gramsetron(Gra) combined with dexamethasone(DEX) on preventing postoperative nausea and vomiting(PONV) after umbilical one-trocar laparoscopic appendectomy(UOTLA)in children.Methods Sixty children requring UOTLA under general anesthesia were allocated randomly into 3 groups,and each of group included 20 children.Group saline(C),group Gra 40 ?g/kg,group Gra 40 mg/kg and group DEX 0.2 mg/kg(Gra+DEX) were respectively injected intravenously after appendectomy performed outside the abdominal cavity during the operation.Results The incidence of PONV decreased significantly in group Gra and group Gra+DEX than that in group C(Pa

BACKGROUND: Child neglect is one of the most important aspects of child abuse and has become a serious problem influencing children's development all over the world. Therefore, it is necessary to nderstand the current conditions and influential factors of child neglect and establish effective evaluation methods in China.OBJECTIVE: To investigate the current conditions and influential factors of urban child neglect in Suzhou city.DESIGN: A comparative analysis of sampling survey.SETTING: Department of Children's Health Care, Soochow University Affiliated Children's Hospital.PARTICIPANTS: This study was conducted in one public kindergarten and two private kindergartens of urban areas in Suzhou city between March and October 2003. Totally 370 children (196 boys and 174 girls) aged between 4 to 6 years old without obvious physical and mental diseases were voluntarily investigated, and they had been living with the person that answered the questionnaire regarding to child neglect in China in the recent 1 year. METHODS: Questionnaire regarding to child neglect in China was sent to each volunteer.he questionnaire evaluates the current conditions of child neglect by 20 items of 5 aspects including physical care (buy fruit for child, consider the nutritional need of child, care about the body height and body mass of child), emotional care (chat with child, play and tour with child, encourage child to do the things whatever they can, listen to what the child feels, introduce the child to the guests when they come to your home, let him or her stop immediately when child cries, care about the activities in kindergarten), education care (buy books for child, tell stories to child, recognize and correct the bad behavior of child, answer the questions asked by child seriously), safety care (protections against fire and electricity at home, warn child against matches, knives and other sharp things, teach child how to cross the road) and health care (prepare the common medicine for child at home, urge child to brush teeth, correct the bad posture). The volunteers were asked to answer each question objectively according to the conditions of child within the recent 1 year, and each question should be answered by selecting only one choice (never, occasionally, usually, always), and anyone should not be omitted. Meanwhile,the educational background and occupation of the parents were also investigated.MAIN OUTCOME MEASURES: The general status of child neglect and the effects of different family educational background and occupation of parents on child neglect were observed. RESULTS: Totally 370 questionnaires were sent out, and 370 qualified ones were recollected, the effective rate was 100%. ① In Suzhou city, the children were given better regards and good care in the aspects of education, medicine and safety, but neglects on children's physical and mental care still exists in a few families. ② Families of different educational background could think highly of the medical care, safety and psychical care of children, but neglects on education and emotional development more commonly existed in the families with the educational background under university. ③ All the families with different occupation could think highly of the medical care, safety and physical care of the children, but education and emotional development were better in the cadre families than in the ordinary families.CONCLUSION: In Suzhou city, the children are given better regards and good care in the aspects of education, medicine and safety, but some bad manifestations of child neglect exist in education and emotion, which are associated with the educational background and occupation of the parents.Further studies about the evaluation, prevention and intervention are needed for child neglect in China.

Objective To investigate the effects of the silence of teratocarcinoma-derived growth factor-1 ( TDGF-1 ) gene on invasion of human pancreatic cancer cell. Methods Three small interfering RNA (siRNA) targeting for TDGF-1 genes (S1, S2, S3 ) were designed and established, then the gene with the best silencing effects was screened. Human pancreatic cancer cell line PANC1 were transfected by siRNA with different concentrations (3. 125, 6.25, 12.5 nmoL/L), the cells without transfection, and simply treated with liposomes were controls. The expressions of mRNA and protein of TDGF-1 were determined by real time PCR and Western blot assay, respectively. The anchorage-independent growth was examined by clon formation in soft agar, and invasion ability was evaluated by boyden chamber model. PANC1 cells with transfection for 48h were injected into the nude mice to evaluate the invasion ability in vivo. Results The expressions of TDGF-1 mRNA and protein of cells transfected by siRNA were decreased in a dose-and time-dependent manner, which were significantly lower than those in liposomes group. Number of colony formation and transmembrane cell were 19.8 ± 2.2 and 49.8 + 2.6 in the control group, and 5.6 + 1.2 and 8. 1 + 1.1 in the 12.5 nmol/L transfection group. The volumes of tumor 4 weeks after transplation in the control group, liposomes group and the 12.5 nmol/L transfection group were (2.228 ± 0.016 ) cm3, ( 2.186 ± 0.028 )cm3 and ( 0.728 ± 0.023 )cm3. Conclusions TDGF-1 gene silence could inhibit invasion ability of human pancreatic cancer cell PANC1.

Objective To investigate the distribution and antimicrobial resistance of gram-negative bacilli isolated from wound specimens of orthopedic patients,and provide reference for the rational use of antimicrobial agents. Methods 682 isolates of gram-negative bacilli were collected from orthopedic department in a comprehensive hospi-tal between January 2011 and December 2013, antimicrobial susceptibility testing results were analyzed. Results The main gram-negative bacilli isolated from wound specimens of orthopedic patients were Pseudomonas aeruginosa (P .aeruginosa)(16.72%),Escherichia coli (E.coli)(15.40%),and Enterobacter cloacae (E.cloa-cae)(12.76%).The detection rates of extended-spectrum beta-lactamase-producing E.coli and Klebsialla pneu-moniae (K .pneumoniae)were 54.29%(57/105)and 31 .43% (22/70)respectively,and mainly distributed in the trauma orthopedic department,accounting for 49.12% and 45.45% respectively.The susceptibility rates of E.coli, K .pneumoniae ,and E.cloacae to meropenem and imipenem were all 100.00%.The susceptibility rates of E.coli and K .pneumoniae to amikacin,piperacillin-tazobactam and amoxicillin/clavulanic acid were all >80%.Suscepti-bility rate of E.cloacae to most antimicrobial agents were 71 .26% -100.00% except for piperacillin(64.37%). Susceptibility rates of P .aeruginosa to most antimicrobial agents were >85% except for cefepime (78.95%)and aztreonam (65.79%).Conclusion Gram-negative bacilli are the most common pathogens in wound infection of or-thopedics patients.In order to use antimicrobial agents rationally and improve clinical treatment effect,it is impor-tant to realize the distribution of pathogens and antimicrobial resistance.

BACKGROUND:Hyaluronic acid or compound betamethasone for treatment of knee osteoarthritis can reduce inflammatory reactions in the joints, relieve pain, and protect the function of the knee joint. OBJECTIVE:To investigate the clinical efficacy of intra-articular injection of hyaluronic acid combined with compound betamethasone on middle-stage knee osteoarthritis as wel as protective effect on joint function. METHODS:A total of 100 patients with II, III knee osteoarthritis were randomly divided into two groups: intra-articular injection of hyaluronic acid (2 mL) as control group and intra-articular injection of hyaluronic acid (2 mL) combined with compound betamethasone (1 mL) as testing group. The intra-articular injection was carried out once a week, totaly for consecutive 5 weeks. Visual analog scale score, Lysholm knee joint score and erythrocyte sedimentation rate were compared between two groups before and after treatment. RESULTS AND CONCLUSION:After treatment, the visual analog scale score, Lysholm knee joint score and erythrocyte sedimentation rate were improved significantly in the two groups as compared with those before treatment (P < 0.05); in addition, these parameters were al better in the testing group than the control group (P < 0.05). No systemic adverse reactions occurred in the two groups. These findings indicate that the intra-articular injection of hyaluronic acid combined with compound betamethasone can reduce inflammatory reactions, relieve pain, and improve joint function.

Objective: To evaluate the effects of functional appliance and multi-bracket appliance on Angle Class II malocclusions coupled with vertical and transversal problems. Methods:Headgear-activator, Herbst appliance and multi-bracket appliance were used to treat 20 patients with Class II malocclusions coupled with vertical and transversal problems aged from 10 to 15 years. The lateral cephalograms were measured with Winceph 8. 0 software and statistical analysis was carried by SPSS 13. 0 software. Results:The sagi-tal, vertical, transversal problems had mainly been resolved at the end of functional appliance treatment. The SNB were increased( P<0. 05), ANB were decreased(P<0. 05), upper posterior teeth were intruded and distally moved(P<0. 05), lower posterior teeth were extruded and mesialy moved(P<0. 05). After multi-bracket appliance treatment, the anterior teeth achieved normal overbite and over-jet. The molar relationship changed from class II to class I. Mandibular plane angle was not clockwise rotated(P>0. 05). The upper and lower dentition were harmony in sagital, vertical and transversal. Conclusion: Functional appliance combined with multi-bracket appliance can be used effectively and conveniently for the treatment of Class II malocclusions coupled with vertical and transversal prob-lems.

OBJECTIVE:To select a technique for preparation of fluconazole for injection and to establish a method for determination of its content.METHODS:The formula was selected on the basis of pH value and species of solution adjuvant.The content was determined by HPLC.RESULTS:The preparation was stable when pH value was between6.5～8.5,and its clarity could be increased by propylene glycol.The detectable concentration of fluconazole showed a good linear correlation in the range of40～200?g/ml.The average recovery was100.37%,RSD=1.37%(n=3).CONCLUSION:The fluconazole for injection prepared by the present technique is stable in quality and the method for content determination is accurate and practicable.