Objective To investigate season distribution,biological typing and drug resistant of Haemophitus in Qingdao Central Hospital.Methods The sputum and throat swab were collected from patients with respiratory tract infection,221 Haemophilus strains were identified and typed by the manual method and MicSCAN4 automatic analyzer,HNID identification plate.Antimicrobial susceptibility was tested by Kirby-Bauer method,and cephalosporins nitrate thiophene paper method was used to detect β-lacta-mase.Results A total of 96 strains of Haemophilus influenzae(1.6%)were isolated,10(10.4%)strains of Haemophilus influenzae were identified as type Ⅰ,31(32.3%)as type Ⅱ,40(41.7%)as typeⅢand 1 5(1 5.6%)as other types.A total of 125 strains Hae-mophitus parl influenzae(2.1%)were isolated,1 5 (12.0%)strains of Haemophilus parl influenzae were identified as type Ⅰ,23 (18.4%)as typeⅡ,69(55.2%)as type Ⅲ and 18(14.4%)as type Ⅳ,other types were not identified.The highest infected rate was in winter.Resistance rate of Haemophilus influenzae and Haemophitus parl influenzae to ampicillin were 40.6% and 44.8%,to tri-methoprim-sulfamethoxazole were 5 1.0% and 66.4%.The prevalence ofβ-lactamase of all strains were 40.6%and 44.8%.But sus-ceptible rates of Haemophilus to cefotaxime,cefuroxime,meropenem,chloramphenicol were over 90.0%.Conclusion The respira-tory tract infections to Haemophilus influenzae and Haemophitus parl influenzae is more frequently found in winter.Type Ⅱ and type Ⅲ are the most prevalent types.The resistance rates of Haemophilus to ampicillin and trimethoprim-sulfamethoxazole are in-creasing,should not be used as empirical treatment of Haemophilus infection.Antibiotics such as cefotaxime,cefuroxime,meropen-em could be chosen for the treatment of respiratory tract infection caused by Haemophilus.

Objective To monitor the resistance of Streptococcus pneumoniae following the 2004 in Qingdao area, and to provide a reasonable basis for clinical application of antimicrobial agents. Methods Collecting respiratory tract, blood, cerebrospinal fluid and other specimens from out-patient and in-patients of some hospital in Qingdao from January 2005 to December 2008. According to the recommendation of NCCLS, antibiotic resistance analysis of 11 kinds of antibiotic to the isolated 231 Streptococcus pneumoniae by micro-agar dilution method, and analysis resistance trends and age differences. Results The results showed that the rate of Streptococcus pneumoniae not sensitive to penicillin is 23. 38% (PRSP: 9.52% , PISP: 13. 85% ) , resistant to cefotaxime is 9. 96% (23/231), resistant to amoxicillin is 12. 55% , resistant to erythromycin is 90. 48% (209/231). PRSP rate of patients younger than 14 years of age 27. 91% (12/43), significantly higher than that of the PRSP rate of adults 5. 38% (10/186). Conclusion The rate of resistant to penicillin Streptococcus pneumoniae increased significantly from 2004, and an increasing trend year by year, the resistance of Streptococcus pneumoniae is also a rising trend year by year. For patients infected low penicillin-resistant Streptococcus pneumoniae in this region, cefotaxime, amoxicillin are preferred drugs.

Objective To evaluate the value of spiral CT and panoramic radiographs in preoperative assessment the jaw condition of patients for dental implantation.Methods 80 patients required dental implant restoration,these patients were Manned by CT and panoramic radiographs in preoperative,then selected suitable patients for dental implant restoration,reconstructed the jaw based on spiral CT data and guided the dental implant design and implanration.Results The spiral CT examination could be more accurate selection criteria for the patients needed dental implant restoration and more accurately showed the situation of edentulous ridge bone.Conclusion The reconstructed of three-dimensional imaging of spiral CT could precisely show the bone situation in the jaw needed dental implant and the important anatomic structures,could effectively assist denture design and the direction and depth of dental implants.was more superiority than panoramic radiographs.

AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.

OBJECTIVE:To systematically evaluate therapeutic efficacies of perioperative application of single-dose and multipie-dose of antibiotics for preventing surgical site infection after closed fractures patients receive incision and internal fixation,and to provide evidence-based reference.METHODS:Retrieved from PubMed,EMBase,CBM,Cochrane Library,CBM,CJFD,VIP and Wanfang database,RCTs about perioperative application of single-dose (trial group) and multiple-dose (control group) of antibiotics for preventing surgical site infection after closed fractures patients receive incision and internal fixation were collected.Meta-analysis was performed using Rev Man 5.3 statistical software after literature screening,data extraction and quality evaluation by improved Jadad.RESULTS:Finally 8 clinical studies were included,involving 3 457 patients.The results of Meta-analysis showed that there was no statistical significance in the rate of surgical site infection [OR=1.02,95％ CI(0.63,1.66),P=0.94)].Cefazolin stadies were included in subgroup analysis according to drug use,and results of Meta-analysis showed that there was no statistical significance in the rate of surgical site infection[OR=1.33,95％CI(0.55,3.20),P=0.52].CONCLUSIONS:Prophylactic application of single-dose of antibiotics is recommended for preventing surgical site infection after closed fractures patients receive incision and internal fixation.

Tendons and bones are connected at the tendon-bone interface to transmit force and exchange biological information. However, the formation of fibrous scars after injury to the tendon-bone interface makes it difficult to recover the original structure during surgery and thus reduces its performance. Therefore, the healing of the tendon-bone interface is a hotspot in sports medicine. Numerous studies have already demonstrated that a variety of molecules and cells participate in the tendon-bone interface reconstruction process, and yet the specific mechanism remains unclear. At present, a great number of studies have been carried out on treatment methods, but clinical treatment are varied with no unification. Therefore, the authors review the advances in the biology and mechanics of healing mechanisms of tendon-bone interface as well as the main methods promoting tendon-bone interface healing, so as to provide references and new ideas for further researches on tendon-bone interface healing.

AIM:To study whether glycyrrhizic acid(GL)can resist the ototoxicity of cisplatin(CDDP)in mice and its molecular mechanism.METHODS:Male C57BL/6J mice were divided into 5 groups:control group,DMSO(5%)group,CDDP(4 mg/kg)group,CDDP+low-dose(50 mg/kg)GL group,and CDDP+high-dose(100 mg/kg)GL group(n=14).Auditory brainstem response(ABR)was used to detect hearing changes of mice.HE staining was used to observe the morphological change of cochlear stria vascular in mice.Evans blue(EB)staining was used to observe the per-meability change of the blood-labyrinth barrier(BLB).Immunohistochemical technique was used to detect the expression and distribution of adhesion protein VE-cadherin and tight junction protein ZO-1 on the cochlear stria.ELISA assay and immunofluorescence technology were employed to detect the expression of tumor necrosis factor-α(TNF-α)and interleu-kin-1β(1L-1β).RESULTS:In CDDP group,ABR waveforms of all frequencies were disturbed,the hearing threshold was significantly increased,and I wave latency was prolonged(P＜0.05).In CDDP+GL group,ABR waveforms of various frequencies were well differentiated,the hearing threshold was significantly decreased,and the latency of I-wave was shortened(P＜0.01).The disordered morphology and more vacuoles in the stria vascularis were observed by HE staining in CDDP group.The GL alleviated CDDP-induced damage in the stria vascularis.In EB staining,CDDP caused an increase in per-meability of BLB(P＜0.01),which was improved by GL treatment(P＜0.01).Immunohistochemical results showed that the expression of VE-cadherin and ZO-1 in CDDP group were decreased(P＜0.01),which was restored in CDDP+GL group(P＜0.01).The ELISA and immunofluorescence results showed that the expression of IL-1β and TNF-α was in-creased after CDDP treatment(P＜0.01),which was restored in CDDP+GL group(P＜0.01).CONCLUSION:The GL alleviates CDDP-induced hearing loss in mice by inhibiting CDDP-induced inflammation and reducing the permeability of BLB.

Objective To investigate the molecular mechanism of Shema Zhichuan Liquid in the treatment of neutrophilic asthma(NA)based on network pharmacology and in vivo experiments.Methods(1)The TCMSP,literature search and Swiss ADME and Swiss Target Prediction databases were used to search and screen the active components and their targets of Shema Zhichuan Liquid.OMIM,GeneCards,DisGeNET and DrugBank databases were used to search and screen NA disease-related targets.The intersection of the active components and NA disease-related targets of Shema Zhichuan Liquid was obtained through the microbiology platform to obtain the potential targets of Shema Zhichuan Liquid for the treatment of NA(common targets).Cytoscape 3.8 software was used to construct the network of"Chinese medicinals-active components-potential targets".The PPI network of potential targets was established by STRING database,and the core targets were obtained by analysing the built-in Mcode plug-in.The Metascape platform was used to enrich the gene ontology(GO),Kyoto Encyclopaedia of Genes and Genomes(KEGG)pathways for the potential targets.(2)BALB/C mice were acclimatised and fed for 1 week and randomly divided into a blank group,NA model group,low-dose group(2.5 g·kg-1)and high-dose group of Shema Zhichuan Liquid(10 g·kg-1),and control group of Dexamethasone(1 mg·kg-1);the NA mouse model was replicated by intraperitoneal injection of sensitizer(OVA+CFA)and nebulized inhalation excitation.OVA/CFA(20 μg OVA+75 μg CFA,0.3 mL)was injected intraperitoneally to sensitize on days 0,7 and 14 respectively,and 5%OVA suspension was nebulized on days 21-30(8 mL each time,40 minutes each time,once a day);1 hour before nebulisation,each group was administered by gastric gavage,and the Dexamethasone control group was administered by intraperitoneal injection once a day.The pathological changes of mouse lung tissue were observed by HE staining;IL-8 content in alveolar lavage fluid was detected by ELISA;mRNA expression levels of NLRP3 and CXCR2 were detected by RT-qPCR;and p-mTOR protein expression levels was detected by immunohistochemistry.Results(1)A total of 826 active component targets and 154 NA disease-related targets were obtained,and 51 potential targets(common targets)for the treatment of NA were obtained from the intersection of the active component and the NA disease-related targets of Shema Zhichuan Liquid.Through the network analysis of"Chinese medicinals-active components-potential targets",quercetin,lignocerotoxin,kaempferol,stigmasterol,naringenin and other key active components were obtained.The PPI network analysis of potential targets yielded 29 core targets,including AKT1,IL6,TNF,EGFR,NLRP3,RELA,MIF,CXCR2,VEGFA,etc..The GO functional enrichment analysis yielded 882 biological process entries,33 cellular component entries,and 61 molecular function entries;KEGG analysis yielded 142 signaling pathways,mainly involving TNF signaling pathway,influenza A signaling pathway,Toll-like receptor pathway,MAPK signaling pathway,mTOR signaling pathway and so on.(2)Results of animal experiments:compared with the blank group,mice in the NA model group showed obvious damage to the airway mucosa,structural disorders,a large number of inflammatory cells infiltration,mucosal congestion,oedema,obvious thickening of the alveolar wall,and narrowing of the alveolar lumen;the level of the inflammatory factor IL-8 in the alveolar lavage fluid was significantly elevated(P＜0.05);the mRNA expressions of NLRP3 and CXCR2 in the lung tissues of the mice were significantly up-regulated(P＜0.01),and the protein expression of p-mTOR was significantly increased.Compared with the NA model group,the structural arrangement of bronchial epithelial cells in the mice in the low-and high-dose groups of Shema Zhichuan Liquid was slightly disordered,with a small amount of inflammatory cell infiltration around the airways and blood vessels,and the congestion and edema of the bronchial mucosa were significantly reduced;the mRNA expression of CXCR2 in the lung tissues of the mice was significantly down-regulated(P＜0.01),and the level of expression of p-mTOR protein was significantly reduced.The IL-8 level in the vesicular lavage fluid of mice in the high-dose group was significantly reduced(P＜0.05);the mRNA expression of NLRP3 in the lung tissue of mice in the low-dose group was significantly down-regulated(P＜0.05).Conclusion The therapeutic effect of Shema Zhichuan Liquid on NA may be achieved through the key active components,such as quercetin,lignocerol and kaempferol,acting on the core targets,such as NLRP3 and CXCR2,and regulating the key signaling pathways,such as the TNF signaling pathway,the MAPK signaling pathway,the Toll-like signaling pathway,and the mTOR pathway.

Objective To explore the effects of oxidative damage and selenium on the apoptosis of articular chondrocytes and the expression of selenoprotein genes.Methods C28/I2 chondrocytes were preincubated for 24 h,using sodium selenite (Na2SeO3) or t-butyl hydroperoxide (tBHP) for 24 h.The experiment was divided into six groups,including control group (C,0.00 mg/L Na2,SeO3 + 0.00 μmol/L tBHP),selenium beforehand protection group (S2,0.10 mg/L Na2SeO3),oxidative damage group (O,150.00 μmol/L tBHP),low dose selenium protection group (OS 1,0.05mg/L Na2SeO3 + 150.00 μmol/L tBHP),medium dose selenium protection group (OS2,0.10 mg/L Na2SeO3 + 150.00 μmol/L tBHP),and high dose selenium protection group (OS3,0.15 mg/L Na2SeO3 + 150.00 μmol/L tBHP).After 24 h,Hoechst 33342 staining method was used to observe apoptosis,mRNA expression of glutathione peroxidase 1 (GPX1),GPX4,deiodinase 2 (DIO2),DIO3,selenoprotein P (SEPP1),thioredoxin reductase 1 (TrxR-1) and selenoprotein W(Sel W) was detected by Real-time PCR,both experiments were done three times.Results Apoptotic rates of C,S2,O,OS1,OS2,OS3 groups [(0.78 ± 0.06)％,(13.61 ± 7.11)％,(92.27 ± 3.44)％,(71.38 ± 5.22)％,(44.31 ± 9.16)％,(72.46 ± 4.69)％] were compared between groups,the differences were statistically significant (F =120.10,P ＜ 0.01).The apoptotic rates of O group was significantly higher than that of C group (P ＜ 0.05);compared to O group,the apoptotic rates of OS1,OS2,OS3 groups decreased significantly (P＜ 0.05),OS2 group was the most obvious.DIO2,SEPP1,GPX1,GPX4,TrxR-1,Sel W mRNA levels were compared in the six groups,the differences were statistically significant (F =24.60,14.53,127.60,30.60,637.10,59.64,P ＜ 0.01).Compared to C group (1.00 ± 0.00),the mRNA levels of GPX1 (0.10 ± 0.05),GPX4 (0.43 ± 0.09),TrxR-1 (0.11 ± 0.05) and Sel W (0.72 ± 0.15) in O groups were decreased significantly (P ＜ 0.05);compared to 0 group,the mRNA levels of GPX1 in OS1 (0.20 ± 0.03),OS2 (0.74 ± 0.10),and OS3 (0.30 ± 0.07) were increased significantly (P ＜ 0.05).Conclusion Down-regulated expression of selenoprotein genes are involved in the regulation process of articular cartilage apoptosis caused by oxidative stress,selenium also has a regulatory role in selenoprotein gene expression in articular chondrocytes.