Objective To establish a real-time quantitative PCR assay for the identification of Campylobacter jejuni in fecal samples.Methods Specific primers of the PCR were designed according to the conserved sequences of Campylobacterjeju-ni,and the real-time quantitative PCR assay was established.150 cases of fecal samples were tested by both culture and PCR methods.With the culture testing results as the reference standard,the sensitivity,specificity,accuracy and repetition of the real-time quantitative PCR were validated.Kappa test was used to estimate the difference between the two detection meth-ods.Results The standard carve of the real-time quantitative PCR assay fitted the equationY=-3.51Log(X)+37.09 (R2=0.996)well.The sensitivity,specificity,and accuracy of the established method were 92.4%,95.8% and 94%,respective-ly.The theoretical detection limit of the PCR method was 102 CFU/ml,and its reproducibility was good (CV<5%).Statisti-cal analysis demonstrated that the results of the two methods were consistent,and the consistent strength was very strong (Kappa=0.88,P<0.05).Conclusion The established real-time PCR method can assay the Campylobacterjejuni in human fecal samples rapidly and accurately.

Objective To establish a real-time fluorescence PCR method to detect the drug resistance genes of pathogenic Campylobacter jejunum in human stool samples,and investigate the relationship between quinoloneantibiotic resistance and the related genes in Campylobacter jejuni .Methods According to the gyrA and gyrB gene sequences that related with the fluoroquinolone resistance in Campylobacter jejuni ,the primers of the PCR method was designed and synthesized.A rapid real-time fluorescence PCR method to detect the drug resistance genes in Campylobacter jejuni samples was established,and the optimum reaction system and conditions were screened through an optimized approach.The developed method was com-pared with the classical drug susceptibility assay.Results It was found in the compared results that,there were 8 inconsis-tent strains of Campylobacter jejuni ,2 of the 8 strains were drug sensitive but contented the drug resistance gene,while 6 strains were drug resistant but had no drug resistant gene.Conclusion The established method can be applied to detect the drug resistance relative genes of gyrA and gyrB in Campylobacter jejuni .There was some correlation between the drug re-sistance representation and its genotype,but this point requires further studies.

Objective To evaluate the efficacy of nucleoside analogues (NAs) antiviral therapy on clinical outcome for hepatitis B virus (HBV)-related primary hepatic carcinoma patients after hepatectomy. Methods The clinical data of 156 HBV-related primary hepatic carcinoma patients after hepatectomy were retrospectively analyzed..According to whether accepted postoperative antiviral treatment, all patients were divided into control group (n = 80)and observation group (n = 76). The serum HBV DNA capacity, recurrence-free survival (RFS)and overall survival (OS)were compared between two groups. Results One week, 1 month, 2 months and 3 months after operation , the serum HBV DNA capacity of observation group was significantly lower than that of control group(P < 0.05). One year, 3 years and 5 years after operation, intergroup comparison of RFS rate of both groups showed statistical significance (P < 0.05) and 1 year, 3 years and 5 years after operation, the difference of OS rate of both groups indicated statistical significance (P < 0.05). Conclusion Standard NAs antiviral treatment for HBV-related primary hepatic carcinoma patients after hepatectomy ,can improve prognosis and prolong survival time. The inhibition the HBV copy active may be its mechanism.

OBJECTIVETo investigate the effects of small interfering RNA (siRNA) targeting YAP on the proliferation and apoptosis of human periodontal ligament stem cells (hPDLSCs).

METHODSSynthesized sequences of siRNA were transfected into hPDLSCs by Lipofectamine™ 2000. The expression of YAP was identified by using real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. Proliferation activity was detected by using cell counting kit-8 (CCK-8). Changes in the cell cycle and apoptosis rate were detected by using flow cytometry. Results were analyzed by using SPSS 19.0, and P < 0.05 was considered statistically significant.

RESULTSExpression of YAP mRNA and protein were significantly downregulated after 48 h of transfection (P < 0.001). No obvious difference was found in the expression levels of YAP protein between 48 and 72 h, thus indicating that siRNA could inhibit the expression of YAP persistently and effectively. Proliferation activity was inhibited, and apoptosis rate was increased. Cell cycle was changed as the proportion of G₁and S phases increased (P < 0.01) and G₂ phase decreased (P < 0.05).

CONCLUSIONKnocking down YAP gene by siRNA could inhibit proliferation activity, induce apoptosis, and change the cell cycle of hPDLSCs. Thus, YAP could regulate the proliferation and apoptosis of hPDLSCs.

Adaptor Proteins, Signal Transducing ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; Humans ; Periodontal Ligament ; drug effects ; Phosphoproteins ; RNA, Messenger ; RNA, Small Interfering ; pharmacology ; Stem Cells ; drug effects ; Transfection

The medical and health organization management information system at grass-root level and platform at county level for data exchange in Sichuan Province were designed and constructed according to the health information exchange service network in Sichuan Province and standard medical CDA file, in order to implement data exchange on the medical and health organization management information system at grass-root level and platform at county level, to insure the basic medical and health service for the public, and to improve their health level.

Objective To provide the evidence for health management decision-making and rational use of drugs grass root clinics by studying their drug prescription rules.Methods The prescribed drugs in clinics of 5 township health centers from September 2012 to September 2014 were retrieved from The Management Information System of Sichuan Grass Root Medical Institutions.Their big data were analyzed using R language.Results The commonly pre-scribed drugs in clinics were vitamin B6, vitamin C and cefixime tablets, which were usually used in combination. Conclusion Health administrative organizations can strengthen their supervision and management of prescribed drugs and promote their rational use in grass root clinics using unified management information system of grass root medical institutions in combination with information technology .

Objective To analyze the expression of triggering receptors expressed on myeloid cells-1(TREM1)in esophageal cancer and its relationship with the overall survival rate of patients,and to explore its effect on the migration ability of KYSE30 cell in e-sophageal cancer.Methods The expression of TREM1 in tumor tissue and its relationship with patient prognosis were analyzed by bioin-formatics and clinical specimens.KYSE30 cells were selected and divided into negative control(NC)group,negative control+lipopo-lysaccharide(NC+LPS)group,interference TREM1 group,interference TREM1+lipopolysaccharide group.Compare the changes in cell proliferation and migration ability of different groups using Transwell method;Western blot was used to detect changes in EMT-relat-ed proteins among different groups;real-time quantitative polymerase chain reaction(RT-qPCR)was used to compare interleukin-1β(IL-1β),interleukin-6(IL-6)and interleukin-8(IL-8)of different groups of cells;overexpression of TREM1 interfered with IL-6,and the changes in EMT-related proteins were compared at this time.Results TREM1 was highly expressed in esophageal cancer tissue and serum,and was associated with lymph node metastasis and overall survival rate in patients;TREM1 can promote the in-vasion and metastasis of KYSE30 cells,and the related mechanism may be by promoting the expression of IL-6,thereby initiating EMT in esophageal cancer cells.Conclusion TREM1 is highly expressed in esophageal cancer tissue,and promotes tumor EMT through up-regulation of IL-6,which is beneficial for the invasion and metastasis of esophageal cancer,leading to poor prognosis in patients.

Objective To explore the therapeutic effect of K8 short wave therapeutic apparatus on facial sensitive skin.Methods 130 patients with facial sensitive skin admitted in our hospital from November 2015 to September 2017 were selected as the subjects.According to the different surgical methods,65 cases in the control group were treated with intensive care cream.On the basis of the control group,65 cases in observation group were given K8 therapeutic apparatus.Then we compared the treatment effect between the two groups.Results K8 shortwave therapeutic apparatus to treat facial sensitive skin can achieve higher total effective rate,reduce the skin area score and and erythema parameters,reduce the skin water loss and improve skin stratum corneum moisture content.After two weeks of treatment,the total effective rate of the observation group (98.46％) was higher than that of the control group (86.15％),the skin area score [(2.28 ± 0.26) piont],the erythema parameter (180.28 ± 29.87) and the epidermis water loss [(9.85 ±2.25)g/(cm2 ×h)] were lower than that in the control group [(4.54 ± 1.14) point,223.87 ± 30.47,(13.02 ± 3.25) g/(cm2 × h)],while the water content of the skin cuticle [(88.75 ± 10.35) ％] was higher than that of the control group [(78.98 ± 9.58) ％] (all P ＜ 0.01).Conclusions K8 short-wave therapeutic apparatus combined with intensive care cream has a better clinical effect and certain advantage in the treatment of facial skin sensitive skin than the use of intensive care cream alone.

Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.

Objective To study the mechanical response of tibial prostheses and the distal tibial cancellous bone after implantation of radial and axial functionally graded materials(FGM)into the ankle joint.Methods Three FGM were used:titanium alloy-bioactive glass composite FGM(FGM-Ⅰ),titanium(Ti)alloy-ideal bone elastic composite FGM(FGM-Ⅱ),and Ti alloy-hydroxylapatite composite FGM(FGM-Ⅲ).A three-dimensional finite element model of total ankle arthroplasty(TAA)was established,and the simulation software ABAQUS was used for secondary development based on Fortra.By changing the volume fraction,the mechanical properties of the FGM tibial prosthesis can be adjusted both axially and radially.The stress distributions of the tibial prosthesis and cancellous bone after FGM axial and radial tibial component implantation in the standing position were analyzed.Results Compared with Ti alloy tibial prosthesis,three kinds of FGM could effectively reduce the stress concentration on the tibial prosthesis,and the overall effect of FGM-Ⅲ tibial prosthesis was better than that of FGM-Ⅰ and FGM-Ⅱ tibial prosthesis;the radial distribution of FGM could effectively reduce the maximum von Mises stress of the prosthesis.For the tibial cancellous bone,the three types of FGM radial tibial prostheses and FGM-Ⅲ axial tibial prostheses could effectively increase the distal stress,thus,relieving the stress shielding on the cancellous bone;the FGM-Ⅲ radial tibial prosthesis was the most effective in improving the stress level of the cancellous bone.Conclusions FGM-Ⅲ radial ankle prosthesis can effectively reduce the stress concentration phenomenon and the stress shielding effect on the prosthesis to prolong its life,with potential application prospects.