Objective To explore the potential of converting the chloramphenicol resistant clinical E. Coli isolates to chloramphenicol sensitive ones by employing external guide sequence(EGS) technique in vitro. Methods Recombinant plasmids with EGScat 1+2 and tetracycline resistant gene, named PAlterl EGScat1+2, was constructed. Routine Cacl 2 method was used to introduce recombinant plasmid into the chloramphenicol resistant clinical isolates E. Coli 4758. Colony PCR was used to test and A600 was used to detect growth rates in liquid and solid culture of various concentrations of chloramphenicol. Results The chloramphenicol resistant clinical isolates E. Coli 4758 grew well in chloramphenicol(35 ?g/ml, 70 ?g/ml, 105 ?g/ml, 170 ?g/ml) plates whereas the transformants tE4758 with PAlter1 EGScat 1+2 failed to grow in these concentrations, which indicated its resistance to the chloramphenicol was reversed. Conclusions EGS molecules are able to convert the drug resistance in clinical E. Coli isolates in vitro.

The gene and the amino acid sequence of the structural and regulatory region of the Pseudomonas aeruginosa with different resistance patterns were analyzed. Six strains with different resistance patterns were selected and the AmpC beta-lactamase was identified. The objective gene fragment was amplified by colonies PCR. The sequences of the PCR-products were analyzed. The DNA sequence of the structural gene ampC and the regulatory genes ampR, ampD and ampE was detected. The 6 strains and the wild-type Pseudomonas aeruginosa are highly homogeneous in structural and regulatory region. Some new mutant points were found.
Bacterial Proteins/*genetics ; DNA, Bacterial/genetics ; Point Mutation/*genetics ; Pseudomonas aeruginosa/enzymology ; Pseudomonas aeruginosa/*genetics ; Sequence Analysis, DNA ; beta-Lactam Resistance/*genetics ; beta-Lactamases/*genetics

To study the resistant mechanism and clinical significance of pseudomonas aeruginosa to beta-lactam antibiotics, the outer membrane permeability rate of 30 P. aeruginosa strains to 5 beta-lactam antibiotics was measured and their production of beta-lactamase and the beta-lactamase genes they carried detected. Furthermore, the relationship between the permeability, beta-lactamase and the clinical effects of beta-lactam antibiotics was observed. By using 14C-penicillin and liquid-scintillant isotope assay, the affinity of penicillin binding proteins (PBPS) was measured and their roles in the resistant mechanism studied. It was revealed that the permeability rate was higher in sensitive strains than in resistant ones (P < 0.05). All strains harbored 1-4 beta-lactamase genes and produced beta-lactamase. Higher permeability rate and higher degree of stability to beta-lactamase indicated better clinical therapeutic effects. The affinity of PBPs changed little without regard to the permeability and beta-lactamase. These results suggested that the permeability of outer membrane and beta-lactamase, but not PBPs, played important roles in the resistant mechanism of P. aeruginosa to beta-lactam antibiotics and affected the clinical therapeutic effectiveness of some patients.
Anti-Bacterial Agents/*pharmacology ; Bacterial Outer Membrane Proteins/metabolism ; Microbial Sensitivity Tests ; Permeability ; Pseudomonas aeruginosa/*drug effects ; beta-Lactam Resistance/*genetics ; beta-Lactamases/metabolism ; beta-Lactams/*pharmacology

To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the morphology of L02/pcDNA3.1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/ pcDNA3.1-HBx; (3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P<0.01) and the apoptosis rate of HepG2/pcDNA3.1-HBx was lower than that of HepG2 cells (P<0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P<0.01), and the expression of XIAP in HepG2/pcDNA3.1-HBx was about 4 times that in HepG2 (P<0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of hepatic carcinoma by regulating the expression of XIAP.

Objective:To observe the effect of Shuganlidanpaishiwan and its different extractive fractions on gallbladder motility of rabbits. Methods:Establishing 100 rabbit animal models,the rabbits were randomly divided into 10 groups:the low and high dose groups of Shuganlidanpaishiwan,methanolic extract,petroleum ether and ethylacetate,negative and positive control groups,to observe the gallbladder motility of every group of animals. Results:The contraction rate of gallbladder on 30th minute after Adm were(9.40?3.36) ,(10.00?2.00) ,(14.00?2.00) on high dose of Shuganlidanpaishiwan group,methanolice extract group and the low dose of petroleum ether group respectively,which were significantly higher than that of control group. Conclusion:Shuganlidanpaishiwan,methanolice extract and petroleum ether extract could improve the gallbladder contractility.

Objective To investigate the role of S100B protein in the pathogenesis of patients with Japanese encephalitis (JE).Methods A total of 45 patients were enrolled in the Second Hospital of Hebei Medical University from August 2013 to October 2013,who were diagnosed as JE on the basis of clinical features and positive IgM antibodies against JE virus measured by enzyme-linked immunosorbent assay (ELISA) from the Center of Disease Control of Shijiazhuang.The JE patients were divided into initial phase group,acute phase group and convalescence group based on the course,mild JE group,moderate JE group and severe JE group based on the severity,MRI-no-lesion group and MRI lesions group based on the imaging findings of JE.Twelve cases with no evidence of infection in central nervous system in the meantime were chosen as control.The S100B protein was measured by ELISA.Results The content of S100B protein in cerebrospinal fluid was as follows:522.76 (393.35,620.37) pg/ml in mild JE group (acute phase group:609.77 (549.27,779.71) pg/ml,convalescence group:420.48 (344.36,453.19) pg/ml),792.09 (705.47,1 108.96) pg/ml in moderate JE group (acute phase group:770.19 (646.31,1 069.54) pg/ml,convalescence group:803.45 (602.90,1 396.84) pg/ml),and 1 021.94 (680.84,1 302.15) pg/ml in severe JE group (acute phase group:981.82 (680.84,1 826.28) pg/ml,convalescence group:989.00 (553.62,1 207.67) pg/ml).The S100B protein content was 561.52 (454.36,814.56) pg/ml,803.45 (602.90,1 104.01) pg/ml,762.22 (594.95,1 044.97) pg/ml,581.76 (442.51,1 069.10) pg/ml in MRI-no-lesion group,MRI lesions group,total acute phase group and total convalescence group,respectively.While in control group,the S100B protein content was 266.71 (205.72,390.05) pg/ml.The contents of S100B protein in moderate JE group,severe JE group,total acute phase group,total convalescence group,MRI-no-lesion group,MRI lesions group were higher than that in control group (H =4.864,5.497,5.075,3.918,2.971,4.981,P =0.000,0.000,0.000,0.000,0.009,0.000).The contents of S100B protein in mild JE group was lower than that in moderate JE group and severe JE group (H =-2.786,-3.514,P =0.032,0.003).Conclusions The level of S100B protein in cerebrospinal fluid is related with the severity,duration and imaging presentation of JE patients.The dynamic monitoring of S100B protein levels is of great significance for assessment of the patients' condition and curative effect.

Purpose To analyze the hemodynamic characteristics of brain stem in patients with basilar artery hypoplasia (BAH) by magnetic resonance perfusion-weighted imaging (PWI).Materials and Methods According to the inclusion and exclusion criteria,51 patients with BAH were selected as the BAH group,and 79 patients without BAH were selected as the non BAH group.All patients were examined by MRI,3D-TOF and PWI,and magnetic resonance angiography was acquired after the three examinations.The regional cerebral blood flow (rCBF),regional cerebral blood volume (rCBV),regional mean transit time (rMTT) and time to peak (TTP) values of pontine area were measured.Results The rCBF value of the BAH group [(17.10±6.52) ml/(100 g · min)] was significantly lower than that of the non BAH group [(29.06± 13.32) ml/(100 g · min)] (P＜0.05);the rCBV value of the BAH group [(1.41 ±0.26) ml] was significantly lower than that of the non BAH group [(2.62± 0.82) ml] (P＜0.05);the TTP value of the BAH group [(6.14± 1.31) s] was significantly higher than that of the non BAH group [(5.39 ± 1.08) s] (P＜0.05);the rMTT value of the BAH group [(20.78±3.48) s] was significantly higher than that of the non BAH group [(19.01 ±2.39) s] (P＜0.05).TTP was the most sensitive index of cerebral perfusion injury,and the incidence of TTP extension was 41.18％ in the BAH group.Conclusion PWI can detect the abnormal cerebral hemodynamics in patients with BAH,which provides the basis for the timely treatment and prevention of irreversible injury in the ischemic area of the brain.

The noninvasive measurement of liver stiffness (LS) was evaluated by transient elastography (FibroScan) and the possible influencing factors from the patients' clinical situations including age, gender, liver inflammation represented by alanine transaminase (ALT) and total billirubin (TBIL) level, HBV replication (HBV DNA loads), portal vein pressure (portal vessel diameter, PVD), splenic thickness (SPT) and body mass index (BMI) were analyzed in patients with chronic hepatitis B (CHB). A total of 466 patients including 31 patients with acute-on-chronic liver failure (ACLF), and 435 patients with chronic hepatitis B (CHB) among which 82 patients were diagnosed with liver cirrhosis (LC) by clinical manifestations and liver B-type ultrasonic inspection were enrolled at Tongji Hospital from April to December 2009. LS was measured by a FibroScan device (EchoSens, France). Simultaneously, ALT and TBIL levels, HBV DNA loads, PVD, SPT and BMI in all patients were also tested. Forty-one healthy volunteers served as controls. The values of LS were correlated positively with ages of CHB patients and significantly higher in males than in females. In patients with BMI>28 kg/m(2) (obesity) and abnormal levels of ALT and TBIL, LS values were significantly increased as compared with those having normal levels of ALT and TBIL. The patients with ACLF had the highest LS value. Furthermore, LS values in the patients with LC were significantly higher than those in patients without LC. It is concluded that noninvasive measurement of liver fibrosis by FibroScan provides an alternative method to evaluate liver fibrosis of patients with CHB. In order to properly illustrate the stiffness value taken by transient elastography, patients' gender should be taken into consideration and it is also suggested to avoid possible influencing factors including liver inflammation (high levels of ALT and TBIL) and obesity (high BMI).

The gene and the amino acid sequence of the structural and regulatory region of the Pseudomonas aeruginosa with different resistance patterns were analyzed. Six strains with different resistance patterns were selected and the AmpC β-lactamase was identified. The objective gene fragment was amplified by colonies PCR. The sequences of the PCR-products were analyzed. The DNA sequence of the structural gene ampC and the regulatory genes ampR, ampD and ampE was detected. The 6 strains and the wild-type Pseudomonas aeruginosa are highly homogeneous in structural and regulatory region. Some new mutant points were found.

To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the morphology of L02/pcDNA3. 1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/pcDNA3.1-HBx;(3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P＜0.01) and the apoptosis rate of HepG2/pcDNA3. 1-HBx was lower than that of HepG2 cells (P＜0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P＜0.01), and the expression of XIAP in HepG2/pcDNA3. 1-HBx was about 4 times that in HepG2 (P＜0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of hepatic carcinoma by regulating the expression of XIAP.