OBJECTIVE To analyze the relationship of Acinetobacter baumannii in Ningbo No.2 Hospital by pulsed field gel electrophoresis(PFGE) and amplified fragment length polymorphism(AFLP).METHODS The sensitivity of 13 kinds antibacterials were analyzed according to American National Committee for Clinical Labotatory 2004′s Standard.The genotypes were analyzed by PFGE and AFLP.The relationship was analyzed with cluster analysis methods.RESULTS Twenty seven strains of A.baumannii were divided into two groups by AFLP method or into four groups by PFGE method.Cluster analysis showed the concordance.CONCLUSIONS Amplified fragment length polymorphism has more clinical practice.

OBJECTIVE To analyze the existence state of drug-resistant gene,integron and Tn21/Tn501 transposon and the relationship of Acinetobacter baumannii in Ningbo No.2 Hospital by genetic mark.METHODS Twenty seven strains of A.baumannii were clinically isolated.Drug-resistant gene,integron,and Tn21/Tn501 transposon were analyzed using polymerase chain reaction(PCR).The relationship was analyzed with cluster analysis methods.RESULTS The positive rates of blaTEM,blaOXA-23 cluster,blaADC,aac(3)-Ⅰ,aac(6′)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sulⅠ,and intⅠ1 were 81.5%,44.4%,85.2%,85.2%,66.7%,81.5%,85.2%,and 85.2%,respectively.The others were negative.The result of multi-gene cluster analysis showed that the clone transmission existed.CONCLUSIONS The clone transmission of A.baumannii exists in Ningbo No.2 Hospital.The outbreak of A.baumannii may be possible because there are more than 3 clone transmission strains.

Objective The study analyse the multi-drug resistance and epidemiological investigation in Acinetobacter baumannii.Methodes Using,multi-drug resistance gene test,three-dimension test,membranes active efflux test studied multi-drug resistance and epidemiological investigation.The genetype were analyzed by Amplified Fragment Length Polymorphism(AFLP).The strains' homology were researched by multi-gene cluster analysis methods.Results Cluster analysis show that the result of drugsensitive test,multi-drug resistance gene test and AFLP are accordant.The study find at least 3 clones by multi-gene cluster analysis methods.AFLP show that all of the multi-drug-resistance of A.baumannii evolutes from a clone.Conclusions The study combined with clinical practice find that ICU in Ningbo No.2 Hospital now is the core of the arises,evolution,transmission of A.baumannii.Strengthening management of ICU is very important to pretect the prevalence of Acinetobacter baumannii in nosocomial infection.

Background Intravitreal injection with triamcinolone acetonide (TA) may cause complications,including increase of intraoculapressure (IOP),cataracand endophthalmitis.Ketorolatromethamine (Ketorolac) inew,lesadverse reactionof non-steroidal anti-inflammatory drug.The action mechanism of Ketorolaisimilato TA.Therefore,Ketorolamay be completely opartly replace Tin the treatmenof retinal edema.Objective The purpose of thistudy wato investigate the effectof Tand Ketorolaon the expressionof aquaporin-4 (AQP4) and vasculaendothelial growth facto(VEGF) in hypoxiretinal Müllecellin vitro and to explore the mechanism of treating retinal edemwith Tand Ketorolac.MethodThe propose of research and use of the animalwere approved by Animal ExperimenResearch Review Committee of Hubei University of Medicine.Twenty eyeof New Zealand albino rabbitwere extracted and the retinal tissue waisolated.The Müllecellwere cultured and passaged using the enzymatidigestion method and Müllecellwere identified using glial fibrillary acidiprotein (GFAP),vimentin and α-smooth muscle actin (α-SMA) by immunofluorescence staining.The hypoxicell modelwere established by culturing the cellin DMEM with 500 μmol/L CoCl2 fo0,6,12,24 hours.The cellof hypoxifo24 hourwere divided into normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L To50,100,200 mg/L Ketorolagroups.Corresponding drugwere added into the medium in the differengroups.The expressionof AQP4 mRNand VEGF mRNin Müllecellwere detected by semi-quantitative reverse transcription PC(RT-PCR).ResultThe cellgrew well and reached 80％ confluence with the irregulashape and ovoid nuclei 14-15 dayaftecultured.More than 95％ primary cellshowed positive reaction to GFAP,vimentin and α-SMA.The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significantly differenin varioutime point(AQP4 mRNA:F=18.70,P＜0.01 ; VEGF mRNA:F =53.20,P＜0.01),and those of 6,12 and 24 houraftecultured with CoCl2were increased than those withouCoCl2 (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significandifferenamong the normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L ToKetorolagroup(AQP4 mRNA:F =27.98,P ＜ 0.01 ; VEGF mRNA:F =10.03,P ＜0.01).Compared with the hypoxicontrol group,the expressing levelof AQP4 mRNand VEGF mRNin the Müllecellwere declined in the hypoxia+ 100,200 mg/L Tgroup and the hypoxia+100,200 mg/L Ketorolagroup (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNwere found statistically insignificandifference between hypoxia+ 100 mg/L Tgroup and hypoxia+ 100 mg/L Ketorolagroup,obetween hypoxia+ 200 mg/L Tgroup and hypoxi+200 mg/L Ketorolagroup (P＞ 0.05).ConclusionTand Ketorolacan downregulate the expressionof AQP4 and VEGF in Müllecellundehypoxiconditions,inferring thathey have similamechanism in the impacon AQP4 function in retinal edematoueye.

The optimum conditions of baicalin hydrolysis into baicalein by immobilized beta-glucosidase in a two-phase system was studied and the yield was observed. A two-phase system comprising of sodium acetate buffer and chloroform was determined by comparing the solubleness of baicalein in different solvents and partition coefficient of baicalein in related aqueous-organic two-phase system. beta-Glucosidase was immobilized by the crosslinking-embedding method using sodium alginate as the carrier The optimum reaction temperature, pH value, Michaelis constant, the thermal stability and pH stability were assayed. By comparing the yield of baicalin hydrolysis into baicalein by immobilized beta-glucosidase in two-phase system, the optimum reaction conditions were determined-the optimum reaction temperature, pH value and time were 50 degrees C, 5.0 and 10 h, respectively. The yield of baicalein was 85.28%. Compare with one-phase system, two-phase system had an advantage in reaction rate and yield.
Biocatalysis ; Drugs, Chinese Herbal ; chemistry ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; Flavanones ; chemistry ; Flavonoids ; chemistry ; Hydrolysis ; beta-Glucosidase ; chemistry

Objective To investigate the antitumor effects and the possible mechanisms of dendrit-ic cells co-cultured with cytokine-induced killer cells(DC-CIK)in combination with sorafenib on two lung adenocarcinoma cell lines,A549 cells(harboring KRAS gene mutation)and PC-9 cells(harboring EGFR gene mutation). Methods DC and CIK cells were routinely generated in vitro by stimulating PBMCs isola-ted from lung cancer patients with different cytokines and then co-cultured after a week of culturing. Flow cy-tometry analysis(FCM)was used to analyze the phenotype of DC-CIK cells after 7 days of co-culturing. The 50% inhibitory concentration(IC50 )of sorafenib against tumor cells was detected by MTT assay. The tumor cells were treated with DC-CIK cells alone or in combination with sorafenib. The proliferation of tumor cells was tested by CCK-8 kit and dynamically monitored by real-time cellular analysis(RTCA). Annexin-V/ PI staining was used to examine the apoptosis rates in each group. Real-time fluorescent quantitative PCR and FCM were respectively performed to detect the expression of natural killer group 2 member D ligands (NKG2DLs)at mRNA and protein levels after the treatment with sorafenib for 24 h. Results There was no significant difference between the IC50 of sorafenib against A549 and PC-9 cells after a 24-hour exposure(P﹥0. 05). Compared with the DC-CIK biotherapy,treating the tumor cells with DC-CIK cells in combination with sorafenib significantly inhibited the cell proliferation and increased the total apoptosis rates of tumor cells(P﹤0. 05). Moreover,the inhibition rates to tumor cell proliferation were enhanced along with the in-crease of effect-to-target ratio(E/ T). Compared with the single-factor treatment groups,the normalized cell index(NCI)in the combined treatment group was significantly decreased. Blocking NKG2D could abate the inhibitory effect of DC-CIK cells on tumor cell proliferation(P﹤0. 05). The expression of NKG2DLs(inclu-ding ULBP1,UBLP2 and ULBP3)on tumor cells at mRNA and protein levels were increased to different ex-tent after treating with 5 μmol/ L of sorafenib for 24 h. Conclusion There was no significant different be-tween the inhibitory effects of sorafenib on the proliferation of lung adenocarcinoma cancer cells harboring KRAS or EGFR gene mutation. The antitumor effects of DC-CIK cells combined with sorafenib on lung ade-nocarcinoma cells might be induced by regulating the NKG2D-NKG2DLs pathway and enhancing apoptosis. Moreover,the antitumor effects of the combined treatment were better than those of single-factor treatments.

OBJECTIVETo establish fingerprint database of drug-resistant strain of Acinetobacter baumannii with AFLP genotype.

METHODSTwenty-seven strains of A. baumannii were clinically isolated. The microbial sensitivity to 13 antibiotics was analyzed according to NCCLS 2004. The genotype was analyzed by amplified fragment length polymorphism (AFLP) method.

RESULTSThe results were consistent with those with AFLP database.

CONCLUSIONThe database of AFLP fingerprint may have practical value for identification of drug-resistant Acinetobacter baumannii.

Acinetobacter baumannii ; genetics ; Amplified Fragment Length Polymorphism Analysis ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Databases, Nucleic Acid ; Drug Resistance, Bacterial ; genetics ; Genotype ; Microbial Sensitivity Tests ; Polymorphism, Genetic

OBJECTIVETo observe therapeutic effect of point-through-point acupuncture on cerebellar ataxia after apoplexy and evaluate the safety.

METHODSRandom, parallel control, single blind and multicentral study method was used and 224 cases from 4 hospitals were divided equally into a treatment group and a control group, 112 cases in each group. The treatment group were treated with point-through-point acupuncture and the control group with general needling method. Their symptoms and signs, and the effect on transcranial Doppler's method (TCD) were investigated.

RESULTSThe total effective rate was 93.3% in the treatment group which was better than 77.4% in the control group, with a significant difference between the two groups (P < 0.01), and the point-through-point acupuncture could significantly improve TCD of basilar artery, vertebral artery and posterior inferior cerebellar artery (Vs, Vm, Vd, PI, RI), superior to the control group.

CONCLUSIONThe point-through-point acupuncture has obvious therapeutic effect on cerebellar ataxia after apoplexy and good safety.

Acupuncture Points ; Acupuncture Therapy ; Cerebellar Ataxia ; Humans ; Single-Blind Method ; Stroke ; therapy

OBJECTIVEStudy the risk factors that impact the effectiveness of mass hepatitis B vaccination, and try to amend them in the future.

METHODBased on the national surveillance of hepatitis B, all the 1734 of 1-15 years old children from Hebei Province were enrolled in the present study and they were divided into case and control group according to their sera HBsAg were positive or not.

RESULTSMother sera HBsAg positive and the hospital the children were born and earlier year of birth were risk factors.

CONCLUSIONThe effectiveness of mass neonate hepatitis B vaccination has greatly improved and the future focus should be on finding pregnant HBsAg positive women, and encourage them to give birth in better hospitals, and at the mean time, try to make the neonate hepatitis B vaccination perfect, especially in country areas.

Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; China ; Hepatitis B ; immunology ; prevention & control ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Humans ; Immunization Programs ; methods ; Infant ; Male ; Risk Factors ; Vaccination

ObjectiveTo analyze Echinococcus infection in definitive and intermediate hosts in different zones of Qinghai plateau,Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,and to provideascientificbasisfor developing controlstrategiesagainstEchinococcosisinfection. Methods Echinococcosis infection in definitive hosts,dogs and foxes,was identified by morphological observation; in domesticated and wild intermediate host animals was identified by anatomy and pathology; some of the suspected samples were further identified by molecular biological methods.ResultsStray dogs in different zones of Qinghai plateau were infected with Echinococcus granulosus,the infection rates were 38.71％(300/775),49.60％(124/250),and 9.76％(4/41 ) in Qinghai southem plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant(x2 =25.72,P ＜ 0.01 ).in addition,only Qinghai southern plateau dogs were infected with Echinococcus multiloularis,and the infection rate was 16.04％(98/611).The infection rates of fox with Echinococcus multilocularis were 22.89％(38/166) and 30.77％(12/39) in Qinghai southern plateau and Qilian mountain-Hehuang valley,respectively,and wolves were also found to be infected with Echinococcus granulosus in the same areas.The infection rates of domesticated sheep,yaks,goats and pigs with Echinococcosis were significantly different statistically in those different areas(x2 =82.70,41.82,212.63,194.58,all P ＜ 0.01 ).The infection rates of sheep and yaks were higher[43.43％(5664/13 042),49.47％(2917/5896),52.99％ (887/1674),42.18％ (779/1847),50.70％ (1049/2069),52.90％ (685/1295) ] in three areas.The infection rates of goats and pigs [3.26％ (7/215),0.00％ (0/108)] in Qinghai southern plateau were lower than that of other two areas[ 19.51％(119/610),26.91％(43/1598),47.91％(343/716),21.91％(71/324)].The infection rates of Ochotona curzoniae with Echinococcosis were 6.21％ (243/3910),1.80％ (3/167) and 0.00％ (0/199) in Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant (x2 =18.50,P ＜ 0.01 ).Moreover,wild intermediate hosts of Echinococcosis,such as Microtus fuscus,Lepus oiostolus,Pseudois nayaur,Procapra picticaudata,and Prodorcas gutturosa were found to be infected only in Qinghai southern plateau.ConclusionsHuman is faced with a threat of Echinococcosis infection from various definitive hosts in different zones of Qinghai plateau.And stray dogs are the most crucial factor.The life-cycles of Echinococcus are very complicated in Qinghai plateau.Qinghai plateau is a key area in prevention and control of Echinococcosis infection in China.