AIM: To get large amounts of pure antigens to raise specific antibodies and to perform quantifications.METHODS: CYP2B6 (cytochrome P) cDNA fragments was ligated into BamHI restricted PGEX-3b to generate recombinants PGEX/2B6. We identified recombinants PGEX/2B6 by EcoRI digestion. The expression of fusion proteins were induced by adding isopropyl-thiogalactoside(IPTG). Several clones showed high-level expression of fusion proteins. Insoluble proteins was isolated from the bacteria and the fusion proteins was recovered and purified from a preparative (2mm) SDS-PAGE. The polyarcrylamide gel containing the fusion proteins glutathione S-transferase(GST-2B6) were used to immunize BALB/C mice from which polyclonal ascites fluid was prepared. The purified fusion proteins GST-1A1(GST fusion protein of CYP1A1 cDNA246～386aa expressed in this library ,purified by preparative SDS-PAGE), GST-2B6 were used to test the specificity of 2B6pAb. RESULTS:Fusion proteins constructed between GST and CYP2B6 was expressed in Escherichia coli DH5?. Mouse antibodies are raised against the fusion proteins GST-2B6. 2B6pAb was fond to be specific antibody.CONCLUSION:Recombinant PGEX/2B6 were constructed and purified fusion proteins GST-2B6, and specific 2B6pAb were obtained.

Objective To explore the drug resistance and genotypes in 30 strains of multi-drug resistant Escherichia coli isolated from ICU patients to provide the basis for rational use of antibacterial drugs in ICU patients .Methods The MIC method was a-dopted to analyze the antimicrobial susceptibility of Escherichia coli and the Enterobacteriaceae intergenic repetitive sequence (ERIC)-polymerase chain reaction (PCR) method was adopted to perform the genotyping of multi-drug resistant Escherichia coli screened out by the antimicrobial susceptibility .Results 30 strains of Escherichia coli were isolated from ICU and divided into two genotypes of type A and B2 ,among which 16 strains were isolated in sputum (5 strains of type B2) ,9 strains in blood (7 strains of type B2) ,3 strains in lung lavage fluid and 2 strains in urine (1 strain of type B2) .The drug susceptibility test results showed 13 strains of type B2 and their resistance rates to amoxicillin ,cefuroxime ,piperacillin ,ticarcillin were over 90% .17 strains were type A and their resistance rates to amoxicillin ,cephalothin ,gentamicin ,ticarcillin and piperacillin reached more than 70% .The multi-drug resistance in type B2 was higher than that in the type A .Conclusion Type B2 strain of Escherichia coli shows multi-drug resist-ance and strong pathogenicity and mainly distributes in the blood samples .The use of antibacterial drugs in ICU patients should be reasonably adjusted according to different genotypes of Escherichia coli infection .

Chronologicaol order of acupuncture,as one of the in uential factors of acupuncture therapeutic e ect,has been commonly ignored at present.This article is to explain the rationality and necessity of chronologicaol order of acupuncture through following aspects.One is based on TCM theory,such as the'branch'and the'root',gentleness and urgency,qi and blood,reinforcing de ciency and reducing excess.The other is ndings of modern clinical and experimental researches.

Objective To evaluate the possible roles of IL-4,IL-18 in hepatitis B virus (HBV) infection.Methods Serum IL-4,IL-18 level was measured using ELISA,and serum HBV-DNA content was detected by FQ-PCR.Results There was significant in serum IL-4,IL-18 level between the fulminant hepatitis patients and chronic hepatitis patients or normal controls (P

AimTo establish a model of the injury of the vascular endothelial cell(VEC) in rats and observe the function of Chinese herbs of Li Qi Huo Xue(LQHX) will prevent and treat the VEC injury. Methods30 SD rats weredivided into the control, the model and LQHX groups. The model of the VEC injury was established. It tried to demonstrate the effect of LQHX for the coagula tion and fibrinolysis function of the VEC by the C EC count,t-PA, PAI activity, 6-keto-PGF1αcontent and PAgTmax. Results In LQHX group as compared with those of the medol group, The CEC count was redued obviously(P＜0.01). t-PA activity was increased(P＜0.01), so did the percentage of active t-PA(P＜0.01), but PAI activity decreased (P＜0.05), 6-keto-PGF1α content increased (P＜0.01)α the PAgTmax decreasd (P ＜0.01) Conclution LQHX can enhance the anticoagulation and fibrinolysis activities. It is a more effective measure for the VEC protection.

Purpose To investigate the diagnostic value of high-resolution CT reconstruction techniques on the same slice in hypertrophy of transverse process of the fifth lumbar vertebra (HTPL5V), and to provide a basis for clinical diagnosis and treatment. Materials and Methods Twenty-two cases of clinically diagnosed HTPL5V and 20 normal adults were examined with GE LightSpeed 16-slice spiral CT (36 cases) and Philips iCT 256-slice (6 cases). L5 transverse process and the fifth lumber nerve were reconstructed and observed on the workstations. Results In 22 cases of HTPL5V, there were 26 pseudarthrosis formation and 2 sides with L5 transverse process touching the sacral ala. In 28 sides the iffth lumber nerve traveled through false foramina of the HTPL5V including 6 cases of bilateral compression and 16 cases of unilateral compression. In 21 cases, the nerve was compressed by hyperosteogeny on 27 sides (96.4%) and 1 side due to stenosis (3.6%). On 25 sides (89.3%) the compressed nerves were curved in shaper. There was bulging and/or herniated lumbar disc on 9 sides in 7 cases (32.1%). Conclusion High-resolution CT reconstruction techniques can demonstrate the iffth lumbar nerve of HTPL5V and provide evidence for clinical diagnosis and treatment.

OBJECTIVE:To prevent and reduce the occurrence of hospital onset of infection of fungus in our hospi-tal.METHODS:The case history of patients with fungus infection in our hospital during the period from October2002to September2004were consulted and analyzed.RESULTS:Of the107patients with fungus infection in our hospital,72%of whom were above the age of60;the predilection sites were respiratory tract which accounted for81.3%,digestive which ac-counted for14.0%and urinary tract which accounted for10.3%;the main pathogenic bacteria were Candida albicans which accounted for92.5%of the total.All of the infected people were found to have used broad-spectrum antibiotics in a large quantity and some had used corticoid drugs.CONCLUSION:Rational drug use of broad-spectrum antibiotics and corticoid is an important means to prevent hospital onset of infection of fungus.

Objective To explore the effects of applying microsurgical technique in the one-stage urethroplasty for hypospadias with longitudinal preputial island flaps of dorsum penis. Methods Forty-two hypospadias cases with obvious phallocampsis were reviewed. The patients aged from 1 to 19 years (median, 6.4 years). Among them, 6 cases were of coronary suleus type, 28 cases of pe-nis body type, and 8 cases of penis and scrotum type. All patients went through the operative proce-dures by microscope with four-times magnification and microsurgical instruments. Longitudinal prepu-tial island flaps of dorsum penis was employed for the treatment of 33 cases of these patients, and 9 cases underwent longitudinal preputial island flaps of dorsum penis combined scrotal septal pedicle flaps. Results Thirty-eight cases were cured by one-time surgery (90.5%). One case with urethral fistula after operation was cured by surgical repair of urethral fistula. Two cases with urethral meatus stricture after operation and one with urethral stoma stricture were cured by dilatation. Patients were followed for 9 to 52 months (mean, 27 months). Conclusion Use of microsurgical technique with longitudinal preputial island flaps of dorsum penis in one-stage urethroplasty for hypospadias improved the success rate of surgery with few complications.

Objective To understand the basic situation and the influencing factors of medical expenses of the inpatients in Haikou city for the government to developing a scientific control policy of medical expense or improving the existing measures for controlling the medical expense.Methods Totally 463 inpatients from 10 hospitals were conducted the questionnaire survey and the survey data were analyzed.Results The difference of medical costs constituent existed in different medical institutions and the me-dian of drug costs to the total medical costs in general hospital was 42.13%.30.02% (139/463)of the surveyed inpatients thought that the use of “new techniques,new drugs and new equipments”was the primary cause of medical costs rising.51.19%(237/463) of patients thought that the primary considered factor is “cure”when seeing a doctor.24.84%(115/463)of patients did not known the modification of medical insurance policy.46.00%(213/463)and 28.51%(132/463)of patients thought that the main responsi-bility main body for controlling the medical costs rising is government and hospital respectively.Conclusion Increasing the medical insurance policy advocacy,promoting the reform of medical insurance payment methods,strengthening the dominant position of gov-ernment control medical costs and rationally allocating the medical resources conduce to reduce the medical costs rising.

Objective To investigate the different components of Acorus tatarinowii and Polygala tenuifolia(volatile oil, aqueous extract)on the expression of phosphorylated Tau protein at site Ser396 and Tau-5 in the hippocampus of rats with Alzheimer′s disease(AD). Methods Male Sprague Dawley rats were randomly divided into 8 groups:normal control group, model control group,low-,middle-,and high-dose groups of volatile oil of Acorus tatarinowii and Polygala tenuifolia,and low-, middle-,and high-dose groups of aqueous extract of Acorus tatarinowii and Polygala tenuifolia. The subacute aging model was established by intraperitoneal injection of D-galactose( D-gal). Rats were given different components of Acorus tatarinowii and Polygala tenuifolia(crude drug dosage,0.6,1.2,1.8 g·kg-1 )in experimental groups,and 0.9% sodium chloride solution in normal control group and model control group,by gavage for 28 days.The levels of phosphorylated Tau protein at site Ser396 and Tau-5 were detected in hippocampal tissues by Western blotting and immunohistochemistry. Results The levels of phosphorylated Tau protein at site Ser396 were significantly enhanced in the model control group,as compared with those in normal control group (P<0.01).The relative expression levels of Tau protein Ser396 were 3.83±0.10,3.35±0.01,3.11±0.01,2.75±0.03,2.93±0.01,2.55± 0.07,and 2.23±0.08 in model control group,low-,middle-,and high-dose groups of volatile oil of Acorus tatarinowii and Polygala tenuifolia,and low-,middle-,high-dose groups of aqueous extract of Acorus tatarinowii and Polygala tenuifolia,respectively.Two components of Acorus tatarinowii and Polygala tenuifolia could dephosphorylate Tau protein Ser396 to vary degrees in a dose-dependent manner.The aqueous extract component was slightly better than the volatile oil component,especially in the high-dose group(P<0.01).But Tau-5 did not change significantly after treatment with Acorus tatarinowii and Polygala tenuifolia(P>0.05). Conclusion Acorus tatarinowii and Polygala tenuifolia could promote the dephosphorylation of Ser396 site of Tau protein in the hippocampus of AD rats,with the aqueous extract component having better effects.