Ahigh-resolutionmethodforhumanleukocyteantigen-B(HLA-B)genotypingwasestablished based on the optimized polymerase chain reaction, cloning and sequencing technology. The exon2 and exon3 of HLA-B gene were amplified with primers based on the HLA-B gene sequence. These produced heterozygous alleles were effectively cloned into plasmid DNA based on the principle of plasmid incompatibility, and were followed by bacterial culture. Then Sanger sequencing was carried out and after analyzing the result by software ClustalX2 and IMTG/HLA database comparison, the HLA-B genotype of the samples was achieved. Seven clinical samples were detected, and the results were consistent with those of PCR-SBT genotyping method. The method was cost-effective, high-resolution and it did not require technical software. The use of universal primers simplified the cumbersome design and optimization process of specific primers.

Objective To observe the clinical effect of the treatment of bronchial asthma with the Qingfeiyin and Bailing capsule.Methods 107 cases with bronchial asthma were classified into a control group and a treatment group randomly.The control group was treated with the Ipratropine (40-80 ug for each time)and the treatment group was glven Ipratropine and Qingfeireyin(50ml for each time)plus Bailing Capsule(30pills for each time).One course of treatment was 2months.Results The therapeutic effect of treatment group was significantly better than the control group(P＜0.05).Condusion Qingfeiyin and Bailing capsule has excellent effect in treating bronchial asthma.

Objective To evaluate the feasibility of increasing the perfraction dose in treatment of neoplasms by 3DCRT (three- dimention confornial radiation therapy). Methods From May 1998 to June 2002, the radiation therapy plans of 300 out- cranial neoplasms patients were analysed retrospectively, including 143 patients with chest neoplasms and 157 patients with abdomen neoplasms. The PTV was 7.0 ~ 1 478 cm3, major PTV was encircled by 90 % isodose curve, minor 95 % PTV were encircled by 80 % isodose curve. Prescription dose was 90 % reference point dose, perfraction dose was 5 ~ 10 Gy, a majority of dose was 6 ~ 8 Gy, period of treatment was 5 ~ 15 days with an interval of 0 ~ 1 day. The general dose was given to radical cure dose or appeasement dose. The biological effect increased 10 % ~ 30 %. Results All treatment plans were accomplished and there were not complication which reduced patients' QOL. Conclusions 1.Owing to the f factor, increasing dose of perfraction, shortening general period of treatment and improving radiative biological effect were possible during the 3DCRT. 2. It was suggested that the larger out- cranial neoplasms should be treated by 3DCRT firstly, but precise plan, precise design and precise treatment can not intensely be pursued because of the limit of knowledge. 3. During the 3DCRT for out- cranial neoplasms, 2 ~ 3 times routine radiation therapy dose was secure, credible and effective according to different purpose.

Objective To systematically evaluate the association between protein C and Legg -Calve -Perthes disease.Methods A literature research was performed through PubMed,Embase,Cochrane library,Web of Science,Chinese Biomedical Literature Database(CBM),China National Knowledge Infrastructure(CNKI)and Wan-fang Database from inception to February 201 6 on the association between protein C and Legg -Calve -Perthes disease.According to the Newcastle -Ottawa Scale(NOS)criteria,the quality of studies was evaluated and data were extracted.Meta -analysis was performed with Stata 1 1 .0 software.Results A total of 1 4 articles were included.Twelve articles on protein C and Legg -Calve -Perthes disease in the study group and the control group were compared.The results of Meta -analysis showed that there was no significant difference in protein C levels between the study group and the control group[odds radio(OR)=1 .41 ,95% confidence interval(CI)(0.87,2.28),P =0.1 47];five articles on protein C and the white race of Legg -Calve -Perthes disease between the study group and the control group were com-pared,The results of Meta -analysis showed that there was no significant difference in protein C levels between the whiteskin patients′group and the control group[OR =0.61 2,95%CI(1 .83,7.29),P =0.61 2];three articles on pro-tein C and the yellow race of Legg -Calve -Perthes disease between the study group and the control group were com-pared,and the results of Meta -analysis showed that there was no significant difference in protein C levels between the yellow skin patients group and the control group[OR =0.59,95%CI(0.05,6.72),P =0.080].Conclusion There is no significant correlation between protein C and Legg -Calve -Perthes disease.

Pyrosequencing is one of the important genetic polymorphism detection methods currently, but the complicated pretreatment procedure limits its application in clinical test. To simplify the whole process of pyrosequencing, on the basis of the linear_after_the_exponential_polymerase chain reaction ( LATE_PCR) , we improved the primer design method of LATE_PCR, increased the length and the concentration of the excess primer, applied direct amplification technology with whole blood, and established a whole blood_imLATE_PCR method based on common rTaq polymerase and “HpH Buffer” ( High pH buffer ) . The amplification system was optimized, and the influences of blood anticoagulant and the amount of whole blood template were investigated. The single stranded template for the pyrosequencing was obtained by PCR amplification using a single tube in one_step process, and the alcohol dehydrogenase gene polymorphisms of 24 clinical blood samples were then detected successfully. The results could be used to guide clinical individualized medication. The genotypes of ADH1B locus of 24 samples were 6 cases of AA homozygote, 14 cases of AG heterozygote, and 4 cases of GG homozygote. The genotypes of ADH1C were 20 cases of GG homozygote, 4 cases of AG heterozygote, and no cases of AA homozygote.

Objective To improve the orotracheal intubation verifying technique and reduce the complication of backflow in rat experiment.Methods A new position evaluation of anti-backflow device was designed and made of safety IV catheter and closed IV catheter system.60 adult male Sprague Dawley rats 216±20 g were randomly assigned to two groups: group A (n=40) for verifying placement, group B (n=20) for anti-backflow test.Group A was further divided into group A1 using self-designed positioning device, group A2 using aerosol, group A3 taking cotton fiber for positioning judgment.The group B was divided into two subgroups, B1 and B2, counting escaped bubbles as a means of positioning observation, the difference is that group B1 using frustum of a cone shape anti-backflow device, while the group B2 using common airway tube.Routine endotracheal intubation was performed to observe and record the time of positioning, the location of exhalation phase, and the length of inspiratory phase countercurrent water column.The group A1 further performed tracheotomy under direct vision clearly to confirm the anatomic positioning status.Results During the exhalation cycle,three or more bubbles were observed to escape continuously, indicating that the intubation tube was properly placed and open in the airway.Positioning time: It took 1.75±1.02 respiratory cycles in group A1,3.30±0.95 respiratory cycles in group A2 and 4.10±0.99 respiratory cycles in group A3 to complete the assessment the positioning status.There was no statistically significant difference between groups A2 and A3 (P> 0.05).The time needed for group A1 was significantly shorter than that of groups A2 and A3 (P < 0.01).The longest countercurrent water column length in group B1 was 3.23±0.53 cm, and 8.48±1.01 cm in the group B2.Conclusions The new designed anti-backflow positioning evaluation device is a simple and convenient appliance to evaluate the location of orotracheal intubation in rat experiment.It can effectively improve the positioning efficiency and has practical application value.

Objective To develop perfluorocarbon lipid particles and investigate their basic properties,and target them to human hepatocellular carcinoma cells in vitro by hepatoma monocolonal antibody HAb18 with avidin-biotin interaction.Methods Rotary evaporation and high pressure homogen were used to prepare perfluorocarbon lipid particles, and the appearance and distribution of them were investigated by microscope and electron microscope, the concentration and the size and electric potential were detected.The biotinylated monoclonal antibody HAbl8 was prepared, then the biotinylated degree of the antibody was determined.The biotinylated perfluoroearbon lipid particles labelled with NBD were prepared and targeted to human hepatocellular carcinoma cells in vitro with avidin-biotin interaction.Results These perfluorocarbon lipid nanoparticles were uniform and stable,and the mean diameter of them was 171.9 nm.Hepatocellular carcinoma cells were surrounded by the biotinylated particles labelled with NBD.Conclusions A steady perfluoroearbon lipid particles were prepared and the biotinylated particles can be targeted to hepatocellular carcinoma cells with avidin-biotin interaction.

The effects of dietary supplementation with Clostridium butyricum on growth performance and humoral immune response in Miichthys miiuy were evaluated. One hundred and fifty Miichthys miiuy weighing approximately 200-260 g were divided into five groups and reared in 15 tanks with closed circuiting culture system. The animals were fed 5 diets: basal diet only (control) or supplemented of the basal diet with C. butyricum at doses of 10(3) (CB1), 10(5) (CB2), 10(7) (CB3) or 10(9) (CB4) CFU/g. Compared with the control, the serum phenoloxidase activity was significantly increased by the supplementation (P<0.05), acid phosphatases activity was increased significantly (P<0.05) at the doses of 10(9) CFU/g. Serum lysozyme activity peaked at dose of 10(7) CFU/g and in the skin mucus at dose of 10(9) CFU/g. Immunoglobulin M level in the serum and skin mucus was increased except at dose of 10(3) CFU/g (P<0.05). The growth at the dose of 10(9) CFU/g was higher than that of the control (P<0.05). It is concluded that supplementation of C. butyricum can mediate the humoral immune responses and improve the growth performance in Miichthys miiuy.
Animal Feed ; microbiology ; Animals ; Antibody Formation ; physiology ; Clostridium butyricum ; immunology ; Dietary Supplements ; microbiology ; Fishes ; growth & development ; immunology ; Probiotics ; administration & dosage

Adult ; Carcinoma, Squamous Cell ; etiology ; pathology ; Humans ; Lichen Sclerosus et Atrophicus ; complications ; pathology ; Male ; Penile Neoplasms ; etiology ; pathology ; Time Factors

Objective To clone the cDNA encoding human HMGB1, express it in E. coli, and identify its biological activity. Methods Human HMGB1 cDNA was amplified by RT-PCR and cloned into vector pUC19. After sequence analysis, the cDNA was ligated into prokaryotic expression vector pQE-80L and induced by IPTG to express HMGB1. The protein was purified with Ni~(2+)-NTA chromatography and polymyxin B affinity column. To identify the function of purified protein, the product was co-cultured with THP1 cells. Results Recombinant expression plasmid pQE-80L/HMGB1 was constructed successfully. After purification, the protein purity reached 96%. The recombinant HMGB1 stimulated THP1 to secrete TNF-? . Conclusion The highly purified HMGB1 was obtained successfully, which showed biological activity. These results lay the foundation for further research on the function of human HMGB1.