Objective: To obtain the full-length of platelet-activating factor acetylhydrolase (PAF-AH) gene and express it in prokaryocytes. Methods: The full-length PAF-AH gene was amplified from peripheral blood mononuclear cells (PBMCs) by RT-PCR and then cloned into the expression vector pGEX-4T-3. The PAF-AH-GST was expressed in E. coli BL21 and purified with GST purifying system. The products was identified by SDS-PAGE and Western blot, and their enzymatic activities were detected. Results: Sequence analysis indicated that the sequence of the obtained PAF-AH gene was identical with that in the GenBank. SDS-PAGE and Western blot confirmed the successful expression and purification of PAF-AH, with the molecular weight as expected. Western blotting showed that the purified protein could be recognized by specific antibodies and had immune activities. Enzymatic activity detection confirmed that the protein could specifically hydrolyzing PAF. Conclusion: Recombinant PAF-AH protein with biological activity has been efficiently expressed in E. coli in the present study.

Objective To observe the effect of electroacupuncture on the motor function of acute stroke patients received early rehabilitation.Methods 75 acute stroke patients were divided into the electroacupuncture group(38 cases) and control group(37 cases).All patients of the two groups were given clinical treatment and early rehabilitation,the patients in the electroacupuncture group were added with electroacupuncture treatment.The treatment course was 20 days.The motor function of all patients was evaluated by Nerve Function Insufficient(NFI),Fugl-Meyer Assessment(FMA),and Modified Barthel Index(MBl) before and after treatment.Results The motor scores of all patients in the two groups significantly improved after treatment(P<0.01).While, the scores of the cases in the electroacupuncture group were obviously higher than that of the control group(P<0.01).Conclusion The effect of early rehabilitation combined with electroacupuncture on acute stroke patients is better than only early rehabilitation.

To investigate the role of malnutrition on expression of thymus melatonin receptor(MR), rats were fed with low protein diet to induce an experimental animal model of malnutrition. MR was detected by radioligand binding assay. In malnourished rats before pregnancy and those during pregnancy, the thymus development in their offsprings was retarded, and the expression of MR was lower than that of the controls. After fed with standard diet,the development of the thymus and the MR in the infant rats improved significantly. The results suggest that malnutrition can retard the development of thymus, so the production of MR in thymus is lowered, and this may affect the physiological role of melatonin.

Objective To analyze the influencing factors of perioperative coagulation dysfunction in neonates with gastrointestinal surgery.Methods Coagulation indexs, including plasma prothrombin time (PT), partial activated thromboplastin time (APTT), thrombin time (TT) and plasma fibrinogen (Fib), were collected retrospectively from June 2002 to April 2016 in gastrointestinal neonatal surgery, as well as perioperative clinical data and the perioperative coagulation dysfunction associated risk factors were analyzed.Results There were 158 cases with abnormal coagulation indexs within 4 hours after operation, including 61 cases of mild abnormalities and 97 cases of obvious abnormalities.Multivariate logistic regression analysis showed that preoperative pneumonia (OR=2.880, 95%CI 1.417-5.852), perioperative low calcium (OR=2.381, 95%CI 1.167-4.857) and invasive puncture catheter (OR=2.490, 95%CI 1.299-4.773) were independent risk factors for neonatal coagulation index abnormality within 4 hours after surgery (P<0.05).Conclusion In the perioperative management, pneumonia should be actively treated, correcting hypocalcemia and choosing concentrations of less than 6.25 U/ml of heparin solution or normal saline careing arteriovenous catheter reduces the occurrence of neonatal postoperative coagulation dysfunction.

BACKGROUND: Fixation of bone fracture is one of the fundamental methods for bone fracture healing. The technique of AO has a lot of defects, such as negative effect induced by "stress dodging". Recently, the technique of CO is praised highly by national and international scholars. OBJECTIVE: To observe the influence of external fixation of small splint on healing of long bone fracture of rabbit, and compare to the internal fixation of steel plate.DESIGN: Randomized and controlled animal trial. SETTING: Research Institute of Orthopedics, Hubei College of Traditional Chinese Medicine.MATERIALS: The experiment was performed at the Laboratory of Orthopedics, Hubei College of Traditional Chinese Medicine from April 2006 to April 2007. Thirty rabbits were randomly divided into small splint fixation group and steel plate fixation group with 15 rabbits in each group. Small splint was self-made of fir-barks with good elasticity, and composed of exterior, interior, front and back splints. The upper part of the splint was wide and the lower part was narrow. We sting an eyelet in the small splint that is used in front and behind part. A hole was drilled in the front and back splints close to the tubercle of tibia. Steel plate was provided by Jiangsu Golden Deer Group (Type HA2.0). METHODS: The standard models of transverse fracture of 3 mm in the meta-infer 1/3 of left tibia were established. In small splint fixation group (SSF group), the fracture was fixed by plaster stone, and 5 days later, replaced by external fixation of small splint. The steel fixation group (SF group) was fixed by steel plate with 4 holes. Animals were executed 14, 24, and 34 days after surgery, respectively. The growth condition of bony callus in fracture sites was observed, and the histomorphology of bony callus and bone cell production during fracture healing was observed. MAIN OUTCOME MEASURES: Macroscopic observation of rabbit tibial bony callus, and histomorphology of bony callus and bone cell formation. RESULTS: In SSF group, the bony callus formed early, and there were plentiful and active osteoblast. Thirty-four days after surgery, bony union was observed in fracture sites. In SF group, there was little fibrous bony callus in the fracture ends 14 days after surgery, accompanied by granulation tissue. Twenty-four days after surgery, sparing cartilage synostosis was observed. On day 34 days, bony callus span the fracture ends, but fracture ends did not connect completely yet. Compared with the SF group, the quantity of bony callus and the speed of fracture healing were superior in SSF group. CONCLUSION: The external fixation of small splint can promote osteoblastic differentiation and proliferation, absorption of hematoma, calcification of the bony callus, and the growth and rebuilding of bone trabecula.

Objective To explore the total treatment effect,layered diagnostic treatment effect,the relationship between effect and site of infection and common side effects of commonly used antifungal agents such as itraconazole,Voriconazole,caspofungin and amphotericin B liposome in invasive fungal infection with hematologic malignancies.Methods The clinical data of 117 cases of patients with hematologic malignancies combining with invasive fungal infection hospitalized in our department from Jan,2005 to Aug,2008 were retrospectively analyzed.Results The total effect rates of itraconazole,Voriconazole,caspofungin and amphotericin B liposome were 69.0%(40/58),77.4%(24/31),64.7%(11/17)and 63.6%(7/11)respectivley(P=0.726).In patients with pulmonary infection,the effect rates of the agents were 63.0%(17/27),85.7%(12/14),50.0%(4/8)and 62.5%(5/8)respectively(P=0.283),which itraconazole group was higher than that of the other agents.The effect rates of the 4 groups have similar rates in patients with liver and spleen candidiasis,fungemia and infections with unknown origin.The rates of 6 weeks survival were 86.2%,87.1%,70.6% and 72.7% respectively.The common side effects of itraconazole and Voriconazole were mainly gastrointestinal reaction and mild hypokalemia.There were few patients showed gastrointestinal reaction(12.1%) and hypokalemia(20.7%)in the former.Some individual showed visual abnormity(9.7%)and external vertebral body symptoms(6.4%).caspofungin showed mild toxic and side effects,which was only gastrointestinal reaction(15.4%),while they were common in amphotericin B liposome group,which were Chill and fever(81.8%),hypokalemia(100%),gastrointestinal reaction(18.2%)and liver damage(9.1%).Conclusion The total effect,layered diagnostic effect and 6 weeks survival rates were similar in itraconazole,Voriconazole,caspofungin and amphotericin B liposome in patients with hematologic malignancies combining with invasive fungal infection.

Objective To study the current status of antibiotic usage in Southwest hospital, so as to improve the rationality in applying antibiotic. Methods A total of 1 668 patients in Southwest hospital during Jan 1 to Dec 30, 2003 were investigated in antibiotic usage and the data were sorted and analyzed. Results Antibiotics were administered in 1 247 cases (74.76%), including only 76 cases (6.09%) based on susceptibility test, 39 cases (3.12%) occurred hospital infection. 297 cases (23.81%) abused antibiotics, among which 131 cases (38.04%) were without definite symptoms or with weak signs, 93 cases (31.31%) used more than two kinds of antibiotics, 24 cases (8.08%) frequently changed antibiotic in the treatment, 27 cases (9.09%) inappropriately prolonged administration and 22 cases (7.40%) improperly combined the same antibiotics. Conclusion Irrational utilization of antibiotics exits in clinical work. Therefore, the rational use of antibiotic based on susceptibility tests should be emphasized.

High intensity focused ultrasound(HIFU) is a technique that ultrasound energy is focused on tumor target to function the treatment through the biological effects including heating effect, cavitation effect and mechanical effect of body to caused cell coagulation necrosis in vivo. Ultrasound contrast agent is one of the most important applicational agent for the treatment of hepatocellular carcinoma(HCC). We discussed in this paper the application of ultrasound contrast agent in the diagnose and focusing of target before the treatment and the assesement of treatment of HCC. Considering the enhancement of cavitation effect, better acoustic environment that enhance biological efficacy of HIFU during the treatment of HCC, ultrasound contrast agent is drawing more attention as an HIFU enhancement agent.

Objective To establish hypoxic-ischemic brain damaged (HIBD) rat model,investigate whether H2S and cystathionine-β-synthase (CBS), the key enzyme for its generation, may be a mediator of electro-acupuncture(EA) stimulation treatment for HIBD. Methods Thirty-two healthy Sprague-Dawley neonatal rats were divided into four groups randomly: sham control group ( n = 8 ); sham + EA group ( n =8); HIBD control group ( n = 8); and HIBD + EA group ( n = 8 ). HIBD rat models were established on their 7-day-old. From the next day ,rats of sham + EA group and HIBD + EA group were electric stimulated 30 min daily for 14 d,BAIHUI and DAZHUI as the acupoints. Control ones were just fixed at the same time,without acupuncture. The rats were sacrificed on the 22 nd day, one day after the treatment course. Cortical H2S concentration was measured by sensitive sulphur electrode assay. The CBS protein expression was measured by western blot analysis and immunohistochemistry for localization. Results The concentration of cortical H2S in HIBD control group was (26. 83 ± 4. 31 ) nmol/mg protein, which was significantly higher than that of sham control group[(22. 78 ± 1.54) nmol/mg protein]( P ＜ 0. 01 ). The H2 S levels in HIBD + EA group and sham + EA group were ( 18.08 ± 2.71 ) nmol/mg protein and ( 18.91 ± 2. 78 ) nmol/mg protein, respectively. Compared with corresponding control group, they were much lower( P ＜ 0. 01 ). The expression of CBS protein in rats with EA stimulation decreased in cortex compared to corresponding control group( P ＜0. 05 ).Conclusion EA can down-resulate H2S/CBS pathway. This may be one of the mechanisms of how EA contributes to the recovery of brain damage.

Objective To observe and analyze the correlation between the DNA methyltransferase-3B (DNMT3B) gene single nucleotide polymorphisms (SNP) rs 1569686 and the susceptibility to colorectal cancer (CRC) in Tianjin Han population.Methods A case-control study was performed.89 CRC patients and 94 normal controls were examined,and Sequenom MassArray system was applied to detect the genotype of rs1569686 in DNMT3B gene polymorphisms.The association results were analyzed of the SNP and CRC.Results The frequencies of genotypes TT,GT and GG at SNP rs1569686 were 74.2％,24.7％ and 1.1％ in CRC patients respectively,which there were no significant differences from that in controls(73.4％,23.4％ and 3.2％; P＞0.05).Conclusion It suggests that the SNP rs1569686 of DNMT3B gene may not be related to susceptibility of CRC in Tianjin Han population.