OBJECTIVETo construct prokaryotic expression systems of Streptococcus pneumoniae ciaH and ciaR genes,and to determine their correlation with drug resistance.

METHODSThe total length of ciaH and ciaR genes was amplified by PCR and their prokaryotic expression systems were established by using routine genetic engineering technique. SDS-PAGE was applied to measure the outputs of target recombinant proteins rCiaH and rCiaR. Rabbits antisera and IgGs against rCiaH and rCiaR were prepared. The resistance to penicillin and cefotaxime of S.pneumoniae strains was examined after CiaH and CiaR were extracellularly and intracellularly blocked by the IgGs.

RESULTThe homogeneity of nucleotide and amino acid sequences of the cloned ciaH and ciaR genes with the reported sequences was 99.9-100% and 100%, respectively. The recombinant bacteria E.coli BL21DE3pET42a-ciaH and E.coli BL21DE3pET42a-ciaR were able to express the target recombinant proteins rCiaH and rCiaR with efficiency. The outputs of rCiaH and rCiaR were 33% and 45% of the total bacterial proteins, respectively. The double immunodiffusion titers of rCiaH antiserum,rCiaR antiserum,rCiaH-IgG and rCiaR-IgG were 1:4,1:4,1:1 and 1:1, respectively. After CiaH was extracellularly or intracellularly blocked by CiaH-IgG, and CiaR was intracellularly blocked by CiaR-IgG, the penicillin-sensitive or cefotaxime-sensitive strains developed resistance to the two antibiotics; but the blocks did not change that of penicillin-resisting or cefotaxime-resisting strains.

CONCLUSIONThe prokaryotic expression systems of S. pneumoniae ciaH/ciaR genes have been successfully constructed in this study. Both the CiaH and CiaR may be involved in penicillin and cefotaxime resistance of the bacterium.

Animals ; Bacterial Proteins ; genetics ; metabolism ; Cefotaxime ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Genetic Vectors ; Penicillin Resistance ; genetics ; Protein Kinases ; genetics ; metabolism ; Rabbits ; Recombinant Fusion Proteins ; genetics ; metabolism ; Signal Transduction ; Streptococcus pneumoniae ; drug effects ; enzymology ; genetics ; beta-Lactam Resistance ; genetics

Objective To evaluate risk factors for central venous catheter-related bloodstream infection(CRBSI)in intensive care unit(ICU)patients by Meta analysis. Methods Literatures about risk factors of CRBSI in ICU pa-tients were retrieved from databases of Cochrane Library,PubMed,Embase,CBM,CNKI,and WanFang Data,RevMan 5.3 software was used for Meta analysis.Results There are 12 literatures in accordance with the inclusion criteria,with a total sample size of 14 422 cases,5 cohort studies and 7 case-control studies,the Newcastle-Ottawa Scale(NOS)scores were 4-6 points. According to Meta analysis,duration of indwelling catheter(WMD,12.25 [95%CI,5.55-18.94]),multi-cavity catheter(OR,3.52[95% CI,1.46-8.52]),femoral vein catheterization (OR,2.44[95%CI,1.34-4.46]),parenteral nutrition(O R,2.47[95% CI,1.18-5.21]),length of stay in ICU(WMD,10.01[95%CI,4.17-15.85]),APACHE II score(WMD,4.46[95%CI,1.25-7.66]),and dia-betes mellitus(OR,1.83[95% CI,1.08-3.09])were significantly different in each group(all P<0.05). Conclusion Risk factors for CRBSI in ICU patients are duration of indwelling catheter,multi-cavity catheter,femo-ral vein catheterization,parenteral nutrition,length of stay in ICU,APACHE II score,and diabetes mellitus. However,due to the limitation of methodological quality of included studies,more strictly designed and large sam-ple prospective studies are needed to verify the result.

BACKGROUNDDespite low morbidity, acute or subacute spinal epidural hematoma may develop quickly with a high tendency to paralysis. The delay of diagnosis and therapy often leads to serious consequences. In this study we evaluated the effects of a series of methods for the diagnosis and treatment of the hematoma in 11 patients seen in our hospital.

METHODSOf the 11 patients (8 males and 3 females), 2 had the hematoma involving cervical segments, 2 cervico-thoracic, 4 thoracic, 1 thoraco-lumbar, and 2 lumbar. Three patients had quadriplegia, including one with central cord syndrome; another had Brown-Sequard's syndrome; and the other seven had paraplegia. Five patients were diagnosed at our hospitals within 3 - 48 hours after appearance of symptoms, and 6 patients were transferred from community hospitals within 21 - 106 hours after development of symptoms. Key dermal points, key muscles and the rectal sphincter were determined according to the American Spinal Injury Society Impairment Scales as scale A in two patients, B in 5 and C in 4. Emergency MRI in each patient confirmed that the dura mater was compressed in the spinal canal, with equal intensity or hyperintensity on T(1) weighted image and mixed hyperintensity on T(2) weighted image. Preventive and curative measures were taken preoperatively and emergency operation was performed in all patients. Open laminoplasty was done at the cervical and cervico-thoracic segments, laminectomy at the thoracic segments, laminectomy with pedicle screw fixation at the thoraco-lumbar and lumbar segments involving multiple levels, and double-sided laminectomy with the integrity of articular processes at the lumbar segments involving only a single level. During the operation, special attention was given to hematoma evacuation, hemostasis and drainage tube placement.

RESULTSNeither uncontrollable hemorrhage nor postoperative complications occurred. All patients were followed up for 1 - 6 years. A marked difference was noted between postoperative and preoperative scales (u = 3.66, P < 0.01). Most patients recovered after therapy, but the recovery of patients treated at our hospitals was superior to that of those transferred from community hospitals (t = 2.95, P < 0.05). Of the patients treated at our hospitals, 4 were cured and 1 was upgraded with scale from A to D, whereas none of those transferred from community hospitals recovered completely, even one remained scale C.

CONCLUSIONSPhysical examination plus MRI is essential to early diagnosis of acute or subacute spinal epidural hematoma. Preventive and curative measures including emergency operation are helpful to the recovery of patients' nerve function.

Acute Disease ; Adult ; Aged ; Female ; Follow-Up Studies ; Hematoma, Epidural, Spinal ; diagnosis ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged

BACKGROUNDIt is difficult and time-consuming for carrying out conventional hand-sewn bilioenteric anastomosis, especially for small bile duct anastomosis and laparoscopic procedure. In order to simplify it, we have developed a novel procedure of sutureless bilioenteric anastomosis with an intraluminal degradable stent. This study aimed to evaluate the feasibility and safety of this technique with cholangioduodenostomy in dog model.

METHODSA patent intraluminal degradable stent tube for sutureless choledochoduodenostomy in dog model was made with polylactic acid in diameter of 3 mm or 4 mm. Thirty-eight dogs were randomly divided into to a stent group (SG, n = 20) and a control group (CG, n = 18). Dogs in the SG underwent sutureless choledochoduodenostomy with intraluminal stent, while the CG underwent conventional choledochoduodenostomy (single layer discontinuous anastomosis with absorbable suture). Dogs of each group were divided into 4 subgroups according to time of death (1, 3, 6, and 12 months postoperatively) to evaluate the healing of anastomosis. Operation time, intraoperative tolerance pressure of anastomosis, rate of postoperative bile leakage, bursting pressure of anastomosis were compared between the two groups. Anastomosis tissue was observed afterwards by pathology evaluation, hydroxyproline content, serum bilirubin, liver enzyme level and magnetic resonance cholangio-pancreatography (MRCP) to assess the stricture.

RESULTSAll procedures were completed successfully. The surgical time of the SG was significantly less than the CG (SG: (19.2 ± 4.3) minutes, vs. CG: (29.2 ± 7.1) minutes, P = 0.000). One bile leakage was occurred in either group. No significant difference of intraoperative tolerance pressure of anastomosis, rate of bile leakage and postoperative bursting pressure of anastomosis, anastomotic stricture, hydroxyproline content, serum bilirubin and liver enzyme level was found between the two groups. MRCP showed no anastomosis stricture and obstruction during months of follow-up.

CONCLUSIONThe technique of sutureless choledochoduodenostomy with a degradable intraluminal stent is feasible and a safe procedure in this dog model.

Animals ; Choledochostomy ; methods ; Dogs ; Female ; Male ; Stents

The yeast fusant ZFF-28, which is high in biomass production and rich in selenium, was constructed after mutagenesis and protoplasts fusion between yeast strains. The total selenium content of ZFF-28 is 1.8 and 1.0 times higher than that of the parental strains Saccharomyces cerevisiae ZY-67 and Saccharomyces kluyveri SZY-198 respectively. Using single factor tests and a L16(4(3) x 2(1)) orthogonal design, the cultivation conditions was optimized as: 50mL culture in 250mL shake flasks in molasses containing 6% sugar and 60microg/mL Se at 28 degree C for 25h at 220 r/min, with the initial pH adjusted to 6.0 - 6.5. Under the optimized conditions, the biomass (dry weight) reached 8.2g/L and the Se content of the cells reached 2050microg/g, with organic and inorganic Se contents being 91% and 9% respectively.
Biomass ; Hydrogen-Ion Concentration ; Saccharomyces ; genetics ; growth & development ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Selenium ; metabolism ; Selenium Compounds ; metabolism

Objective To discuss the technique and technique-related issues of percutaneous kyphoplasty(PKP).Methods The study involved 69 vertebrae in 51 cases of painful osteoporotic vertebral compressive fractures.Under X-ray fluoroscopy monitoring,the fractured vertebral bodies were treated by kyphonplasty with inflatable balloon.The preoperative and postoperative vertebral height and Cobb angle in radiography were measured and analyzed.Results All patients tolerated the procedure well with dramatic pain relief within 72 hours after the procedure.No clinical complication was found.The loss heights of the anterior and mid portion of the vertebral body reduced from 15?4mm and 11?4mm preoperatively to 10? 4mm and 6?3mm postoperatively,respectively.Cobb angle corrected averagely from 22??6? preoperatively to 12??4?.There was significant difference between preoperative and postoperative measures (P

OBJECTIVETo discuss the effect of different positive criteria on the sensitivity and specificity of sputum cytology screening for lung cancer among Yunnan tin miners.

METHODS9223 Yunnan tin miners who received at least one annual sputum cytology screening for lung cancer during the period between 1992 and 1999 were recruited in the study. At time of enrollment, all participants were aged over 40 years old, had at least 10 years of employment as an underground miner and(or) smelter, and had not been diagnosed with malignancy. In our study, a true positive was categorized as having at least one prior positive sputum screening and a diagnosis of lung cancer, while a true negative, by our definition, signified negative sputum examinations and no diagnosis of lung cancer during the follow up time. Based on different positive criteria, sensitivity and specificity of sputum cytology were computed and receiver operating characteristic (ROC) curve analysis was conducted. Z statistic was used to test the differences of the area under ROC based on Hanley and McNeil method.

RESULTSBy the end of following up on December 31, 2001, a total 500 lung cancer cases were diagnosed among 9223 participants: most were squamous cell carcinoma (55.8% (222/398)) and central lung cancers (68.5% (316/461)). 150 lung cancer cases had a previous positive sputum screening result. When positive criteria were taken as grave atypical metaplasia, moderate atypical metaplasia and slight atypical metaplasia, the corresponding sensitivities were 30.0% (150/500), 36.4% (182/500), 53.0% (265/500) respectively; while the corresponding specificities were 98.9% (8628/8723), 95.1% (8611/8723), 77.9% (7033/8723) respectively. The areas under ROC curve according to different positive criterias were 0.645 (95%CI: 0.635 - 0.654), 0.657 (95%CI: 0.668 - 0.667), 0.655 (95%CI: 0.645 - 0.664) respectively. There were no significant differences found in the comparisons between grave and moderate atypical metaplasia, grave and slight atypical metaplasia, moderate and slight atypical metaplasia(Z statistics were 0.780, 0.645, 0.209 respectively, all P values > 0.05).

CONCLUSIONWhile the standard of positive criteria for diagnosis of lung cancer decreased, the sensitivity of sputum cytology screening increased and the specificity decreased. Since there were no significant differences of accuracy for different positive criteria.

Cytodiagnosis ; methods ; standards ; Female ; Humans ; Lung Neoplasms ; diagnosis ; Male ; Mass Screening ; methods ; Sensitivity and Specificity ; Sputum ; cytology

OBJECTIVETo investigate the efficacy and adverse effects of dapoxetine in the treatment of premature ejaculation.

METHODSWe randomly assigned outpatients with premature ejaculation in the proportion of 2:1 to receive 30 mg dapoxetine on demand (n =78) or 50 mg sertraline qd for one month (n = 39). Follow-up was accomplished in 95 cases, 63 in the dapoxetine group and 32 in the sertraline group. We recorded the intravaginal ejaculatory latency time (IELT), clinical global impression of change (CGIC) score, and adverse reactions of the patients and compared them between the two groups.

RESULTSIELT was significantly increased in both the dapoxetine (from [0.87 ± 0.31] to [2.84 ± 0.68] min, P < 0.05) and the sertraline group (from [0.84 ± 0.28] to [2.71 ± 0.92] min, P < 0.05) after medication. Based on the CGIC scores in premature ejaculation, the rate of excellence or effectiveness was 36.5% in the dapoxetine and 37. 5% in the sertraline group, and the rate of improvement was 63.5% in the former and 71.9% in the latter. The incidence rates of dizziness, nausea, headache, and diarrhea were slightly higher (P > 0.05) while those of fatigue, somnolence, and dry mouth significantly higher (P < 0.05) in the sertraline than in the dapoxetine group.

CONCLUSIONOn-demand oral medication of dapoxetine is effective and well-tolerated for the treatment of premature ejaculation.

Benzylamines ; adverse effects ; therapeutic use ; Double-Blind Method ; Ejaculation ; drug effects ; physiology ; Humans ; Male ; Naphthalenes ; adverse effects ; therapeutic use ; Outpatients ; Premature Ejaculation ; drug therapy ; Reaction Time ; drug effects ; physiology ; Serotonin Uptake Inhibitors ; adverse effects ; therapeutic use ; Sertraline ; administration & dosage ; adverse effects ; Time Factors ; Treatment Outcome

To establish a new Western blotting assay for PrP(Sc) detection, we optimized the Western blotting assay with a precipitation procedure of streptomycin sulfate. After digestion with PK, 10% scrapie infected hamster brain homogenates were incubated with 60 mmol/L streptomycin and the precipitated PrP(Sc) was recovered by centrifugation. The enrichment of PrP(Sc) by streptomycin sulfate precipitation was evaluated using Western blotting assay. The results showed streptomycin could bind to PK-treated PrP(Sc), forming high molecular masses, but not influence the glycosylated patterns on SDS-PAGE. Western blot assay revealed that the detective sensitivity of the streptomycin-precipitation PrP(Sc) was remarkably improved. As a sensitive, specific, rapid and flexible protocol for PrP(Sc), the protocol in this study has the potential utility, alone or combined with other techniques, for the detection of low level PrP(Sc) in the specimens from central nerve system, or from peripheral organs or body fluids.
Animals ; Blotting, Western ; methods ; Brain ; metabolism ; pathology ; Brain Chemistry ; Chemical Precipitation ; Cricetinae ; PrPSc Proteins ; chemistry ; isolation & purification ; metabolism ; Prion Diseases ; diagnosis ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity ; Streptomycin ; chemistry

In order to further study the potential interaction between PrP protein and the tubulin and identify the binding region in PrP with tubulin, native tubulin was extracted from rabbit brian tissues, while various recombinant PrP proteins were expressed and purified. The possible molecular interaction between various PrP fusion proteins and tubulin was tested by pull-down and immunoprecipitation assays. Remarkable molecular interaction between the full length PrP and tubulin was observed by pull-down and immunoprecipitation assays. Subsequently, the binding regions within PrP with tubulin were firstly mapped in the aa 23 -- 91 region within N-terminus of PrP peptide. The studies of the association of PrP with tubulin may further provide insight into the unresolved mechanism of active transport of PrP protein in neurons and possible cellular pathways by which prion causes disease.
Animals ; Binding Sites ; genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Humans ; Immunoprecipitation ; Prions ; genetics ; isolation & purification ; metabolism ; Protein Binding ; Rabbits ; Recombinant Fusion Proteins ; genetics ; metabolism ; Tubulin ; metabolism