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 explore the Chinese massage therapy for the treatment of infantile anorexia of spleen and stomach type clinical curative effect.Methods 2 to 6 years old anorexia children treated from September 2010 to September 2012 for general treatment and health care in our hospital were randomly recruited into a Qi deficiency of both spleen and stomach group and a Yin deficiency of both spleen and stomach group with 25 cases in each group.Both groups were treated with chiropractic therapy.On this basis,Qi deficiency of both spleen and stomach group was applied point-massage on Weishu,Pishu,and Zusanli; Yin deficiency of both spleen and stomach group was applied point-massage on Weishu,Ganshu,and Shenshu.Results ① The comparison of disease and syndromes:the total effective rate was 92％ (23/25)and 88％ (22/25) in Qi deficiency of both spleen and stomach group and Yin deficiency of both spleen and stomach group separately,showing no significant difference (x2=0.886,P＞0.05); the total symptoms improving rate was 92％(23/25) and 76％(19/25)in Qi deficiency of both spleen and stomach group and Yin deficiency of both spleen and stomach group separately,the difference showing statistically significant (x2=0.625,P＜0.05).② Symptom score comparison before and after treatment:The main symptoms score and accompanied symptoms score before and after the treatment were [(18.32±2.42) ％,(4.12±0.17) points] and [(16.80±3.77) ％,(5.30 ±0.41) points] in the Qi deficiency group and [(18.18±2.10),(4.45±0.30) points] and [(19.66 ± 3.62)％,(6.01 ± 1.02) points] in the Yin deficiency group.Both groups showed obvious improvements in the main and accompanied symptom scores after the treatment (P＜0.01),but no significant difference between the two groups (P＞0.05).Conclusion The treatment of anorexia due to deficiency of both spleen and stomch with chiropractic and point-massage was effective.

Objective To discuss the early diagnosis and interventional therapy of acute renal infarction.Methods The diagnosis and therapy of 5 cases of acute renal infarction were retrospectively analyzed.There were 4 male and 1 female patients,aged 38-68 years.The symptoms were severe back pain or upper abdominal pain,with or without nausea and vomiting and the time to hospital was 1 to 16 h.Four cases had rheumatic heart disease and 4 cases had artrial fibrillation history.The lab assay results: WBC 8.9-15.8 × 109/L,urine RBC-to + + +,albumin + to + + +,serum creatinin 66-216 μmol/L,serum LDH 350-920 U/L.The doppler ultrasound and CT scan showed large infarction in 3 cases and focal infarction in 2 cases.Percutaneous arteriography,thrombolytic therapy and thrombosuction via catheter were applied promptly.Results The 5 cases were diagnosed and treated timely and effectively.The emboli were cleared.Four patients were followed up for 3-12 months.The creatinin recovered to 53-122 μmol/L,and the blood perfusion of the infarction lesion was satisfactory.Conclusions Early diagnosis and interventional therapy is important for achieving satisfactory recovery of the acute renal infarction.Three dimensional CT angiography could conduce to early diagnosis and follow-up.Thrombosuction via catheter plus thrombolytic therapy is efficient for greater embolus.

OBJECTIVE: To establish a biofilm model of Haemophilus influenzae and observe the effect of ambroxol on biofilm of Haemophilus influenzae and bactericidal action. METHOD: Thirty strains of Haemophilus influenzae were isolated from adenoids of children with adenoidal hypertrophy. Two strains which could build stronger biofilms was selected in a 96-well plate. The effect of ambroxol on biofilms were determined by crystal violet, and the structure of biofilms were observed by scanning electron microscope (SEM). The numbers of viable bacterial in biofilm after ambroxol treatmented determined by plate culture count. RESULT: Through crystal violet assay, significant difference (P < 0.01) between the two group after treatment was found when ambroxol concentration reached at 0.25 mg/ml and 0.49 mg/ml. The biofilms was destroyed by SEM. Ambroxol had the positive effect on bacterial killing by plate culture count,and the effect was in a dose dependent. CONCLUSION Ambroxol could destroy the biofilm of Haemophilus influenzae, and had bactericidal function in vitro.
Ambroxol ; pharmacology ; Biofilms ; drug effects ; Child ; Child, Preschool ; Haemophilus influenzae ; drug effects ; Humans ; Microbial Sensitivity Tests

Objective:There are limited data describing the clinical characteristics and prognosis of culture negative pyogenic spondylitis.The aim of this study was to investigate the treatment,prognosis and clinical characteristics of culture negative pyogenic spondylitis.Methods: A retrospective study reviewed 74 patients who were diagnosed with spondylitis in Peking University First Hospital from January 2010 to December 2015.A total of 27 patients suffered from pyogenic spondylitis.According to the pa-thogenic culture results,the patients were divided into two groups: culture negative group and culture positive group.The clinical characteristics and treatment outcomes between the two groups were compared.Results: The elder were more vulnerable to pyogenic spondylitis,and of the 27 patients,12 patients were female and 15 male.All patients had no history of administration of antibiotics prior to obtaining culture samples.A causative germ was identified in 14/27 patients (51.9％) with Staphylococcus aureus being the most common pathogen.There was no significant difference between the two groups in the patient's age,gender,visual analogue score (VAS),predisposing factor,clinical symptom,sign and spinal segment (P>0.05).Erythrocyte sedimentation rate (ESR) (P=0.056) and C-reactive protein (CRP) (P=0.040) of culture negative group were lower in contrast to culture positive group.The incidence of vertebral abscess in culture negative group was higher than in culture positive group (P=0.046).After treatment,ESR dropped almost equally in both groups,and CRP dropped faster in the culture positive group (P=0.192).At last,there was no significant difference between the two groups in hospital stay,pain relief,open debridement operation rate,and recurrence rate of infection.Conclusion: ESR and CRP of the culture negative patient were lower than those of the culture positive patient,and the incidence rate of paravertebral abscess was higher than that of the culture positive patient.After administration of antibiotics,there was no significant difference between the two groups in duration of antibiotics,open debridement operation rate and recurrence rate of infection.So,culture negative may not necessarily be a negative prognostic factor for pyogenic spondylitis.However,we should watch out for the drug resistant bacteria or double infection,due to the long term use of wide-spectrum antibiotic in culture negative patients.

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 .

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.

BACKGROUND:Percutaneous vertebroplasty is a minimal y invasive treatment for spinal metastasis tumor, but the mechanism of the injected polymethylmethacrylate (PMMA) bone cement in the treatment of tumor is not ful y understood. OBJECTIVE:To explore the cytotoxicity of PMMA bone cement and its monomers on tumor cel s. METHODS:PMMA extracts in the wire drawing and curing periods and different mass concentrations of monomer dilutions were co-cultured with human lung cancer cel s spc-A1. The cel morphology was observed using inverted microscope, the absorbance (A) values were detected by cel counting kit-8 assay, the relative growth rate was calculated, and the toxicity of PMAA bone cement and its monomers was evaluated at 1 and 3 days of culture, respectively. RESULTS AND CONCLUSION:At 1 day of culture, the absorbance values in the 1 and 10 g/L groups were significantly lower than that in the negative control group (P<0.01). The absorbance values in the 1 g/L, 10 g/L, 100 mg/L and wire drawing extract groups were significantly lower than those in the negative control group at 3 days of culture (P<0.05). The relative growth rate in the 1 and 10 g/L groups was 26%-29%, and the level of toxicity was grade 4 after 1-day culture;at 3 days, the relative growth rate was decreased to 12%-16%, and the level of toxicity was grades 4-5. After 3-day culture, the level of toxicity reached to grade 2 in the 100 mg/L and wire drawing extract groups. These results indicate that PMMA bone cement in wiredrawing period and its monomers exert toxic effects on tumor cels.

OBJECTIVE: To investigate the effect of estrogen on expression of the cysteine-rich secretory protein containing LCCL domain 2 (CRISPLD2) in myocardium of lipopolysaccharide (LPS)-induced mice model of sepsis. METHODS: Totally 12 female and 12 male Balb/c mice of specific pathogen-free (SPF) level with 7 weeks were served as objectives. The female and male mice were randomly divided into model groups and control groups, respectively, with 6 mice in each group. The model of sepsis was reproduced by intraperitoneal injection of 10% LPS (5 mg/kg), and the mice in control groups were injected with the same volume of normal saline. The general condition of mice during experiment was observed at 24 hours after injection. All the mice were sacrificed and the heart was harvested after collecting the whole blood. The concentration of estrogen in serum was determined by double antibody sandwich enzyme linked immunosorbent assay (ELISA). The myocardial tissue homogenate was prepared at the same time, and the total protein was extracted. The expression level of CRISPLD2 was determined by Western Blot. Pearson correlation analysis was used to analyze the bivariate correlation. RESULTS: All of the experimental mice survived at 24 hours after injection. The mice in the model groups showed disorder and gray signs of body hair, with diarrhea and decreased appetite. No significant abnormality was observed in the control groups. There was no significant difference in the body weight or concentration of estrogen in serum between model and control group of both female and male mice [body weight (g): 24.6±1.8 vs. 24.5±1.3 in male mice, 18.0±0.8 vs. 17.5±1.1 in female mice; estrogen (ng/L): 11.93±2.59 vs. 12.17±3.87 in male mice, 28.20±5.75 vs. 29.82±6.10 in female mice, all P > 0.05]. There was no statistical difference in the expression of CRISPLD2 in myocardium between male control mice and female control mice (gray value: 1.02±0.19 vs. 1.00±0.11, P > 0.05). No significant difference in the expression of CRISPLD2 in myocardium was found between female sepsis mice and female control mice (gray value: 1.05±0.13 vs. 1.00±0.11, P > 0.05). The expression of CRISPLD2 in myocardium of male sepsis mice was significantly lower than that of male control mice (gray value: 0.29±0.08 vs. 1.02±0.19, P < 0.01), and it was significantly lower than that of female sepsis mice (P < 0.01). It was shown by correlation analysis that the expression level of CRISPLD2 in myocardium of sepsis mice was significantly correlated with serum estrogen concentration [R² = 0.736, 95% confidence interval (95%CI) = 0.560-0.960, P < 0.001]. CONCLUSIONS In female mice with sepsis, the expression of CRISPLD2 is comparable to that of female healthy mice. It is suggested that estrogen can maintain the expression of CRISPLD2 in LPS-induced septic mice at the normal level.
Animals ; Cysteine ; Estrogens ; Female ; Lipopolysaccharides ; Male ; Mice ; Myocardium ; Sepsis