Objective To construct eukaryotic expression vector of human antimicrobial peptide LL-37 pPIC9-LL-37, and transform the plasmid pPIC9-LL-37 into P.pastoris GS115 to obtain the recombinant P.pastoris strains.Methods The full-length of antimicsobial peptide LL-37 gene was artificially synthesized by overlap extension method and was fused to pPIC9 and then the fused plasmid was transformed into E.coli DH5?.After analysis by PCR and sequencing,the plasmid pPIC9-LL-37 was transformed into P.pastoris.The colonies exhibiting the phenotype of His+Mut+ or His+Mut-were screened by means of MM and MD plates and the insertion was confirmed by PCR.Results The results of PCR and sequencing confirmed that the LL-37 gene was correctly inserted into pPIC9. The colonies of 10 His+Mut+ and 9 His+Mut-were obtained by means of MM and MD plates screening and were confirmed by PCR.Conclsion The recombinant P.pastoris strains containing LL-37 were successfully obtained.

Objective To study the relationship between clinical diagnosis and children ventricular premature beat(VPB) electrocardiogram location and morph.Methods Both organic heart disease and without organic heart disease relationship with 109 cases of children ventricular premature beat electrocardiogram location and morph were retrospectively analyzed.Results Children ventricular premature beat location shows that organic heart disease mostly results from left ventricle, without organic heart disease often comes from right ventricle. There was significant difference between above two groups (? 2=37.25 P

Objective To investigate the expression of pro-inflammatory cytokines in lumbar dorsal root ganglions (DRG) of rats model of high-dose fentanyl induced hyperalgesia. Methods 64 male SD rats were divided into 2 groups (n = 32), fentanyl group and normal saline (NS) group. The rats were injected with fentanyl (60 μg/kg) or NS 4 times in total subcutaneously with a 15-minute interval. Mechanical and thermal nociception were measured via the tail pressure test (tail flick thresholds, TFT) and paw withdrawal test (paw withdrawal latency, PWL) at 1 day before, at 1, 2, 3 and 4 hour and on 1 ~ 7 day after administration. 4 rats were sacrificed and the lumbar DRG were harvested to analyze the expression of PGE2 , IL-1β, IL-6 and TNF-αvia ELISA. Results There were no significant changes of TFT, PWL and the expression of pro-inflammatory cytokines in DRG compared to baseline of rats in NS group. The value of TFT , PWL in fentanyl group were above the baseline at the 1 ~ 4 hour and below the baseline at 1~3 day after fentanyl injections. PGE2 , IL-1β, TNF-α and IL-6 increased on 1,3,5,7 day after fentanyl injections significantly. Conclusions High-dose fentanyl induced significant hyperalgesia and up-regulation of pro-inflammatory cytokines in DRG. The expression pro-inflammatory cytokines peaked later and were more protracted than the change of behavior test and show no direct relationship between the two.

The study aimed to establish a population pharmacokinetic/pharmacodynamic (PPK/PD) model of warfarin. PCR-RFLP technique was used to genotype the CYP2C9 and VKORC1 polymorphisms of 73 patients. RP-HPLC-UV method was used to determine the 190 plasma concentrations of warfarin. Application of NONMEM, the clinical information and 263 international normalized ratio (INR) monitoring data were used to investigate the effect of genetic, physiological, pathological factors, other medication on clearance and anticoagulant response. The final model of warfarin PPK/PD was described as follows: CL = θCL · (WT/60)θWT · θCYP · eηCL (if CYP2C9*1/*1, θCYP = 1; if *1/*3, θCYP = 0.708); EC50 = θEC50 · θVKOR · eηEC50 (if VKORC1- 1639AA, θVKOR = 1; if GA, θVKOR = 2.01; V = θV; K(E0) = θK(E0); Emax = θEmax; E0 = θE0 · eηE0. Among them, the body weight (WT), CYP2C9 and VKORC1 genotype had conspicuous effect on warfarin PK/PD parameters. The goodness diagnosis, Bootstrap, NPDE verification showed that the final model was stable, effective and predictable. It may provide a reference for opitimizing the dose regimen of warfarin.
Anticoagulants ; pharmacology ; Body Weight ; Cytochrome P-450 CYP2C9 ; genetics ; Genotype ; Humans ; International Normalized Ratio ; Nonlinear Dynamics ; Polymorphism, Genetic ; Vitamin K Epoxide Reductases ; genetics ; Warfarin ; pharmacokinetics

OBJECTIVETo explore methods and clinical effects of composite external fixtor in treating adult patients with unstable middle 1/3 of clavicular fractures.

METHODSFrom March 2008 to March 2011,36 patients with unstable middle 1/3 of clavicular fractures were treated with composite external fixtor. There were 24 males and 12 females, aged from 18 to 55 years old with an average of 43 years old. Twenty cases on the left side and 16 cases on the right side. Time from injury to operation was 2 to 6 days (averaged 3.5 days). According to Robinson classification, there were 7 cases with type 2A2, 18 cases with type 2B1, and 11 cases with type 2B2. No vessels and nerve damage occurred before opreation. The clinical effects were evaluated according to Neer scoring.

RESULTSAll cases were followed up from 6 to 12 months with an average of 8 months. The mean Neer score was 88.3 +/- 6.2, which included pain 31.6 +/- 3.2, functional score 25.7 +/- 2.2, range of motion score 21.1 +/- 1.7, and anatomy score 8.8 +/- 0.8. There were 22 cases in excellent, good in 13, fair in 1. Two cases occurred pin tract infection.

CONCLUSIONComposite external fixtor is an optional method in treating unstable middle 1/3 of clavicular fracture, and can obtain a good clinical effects.

Adolescent ; Adult ; Bone Nails ; Clavicle ; injuries ; surgery ; External Fixators ; Female ; Fracture Fixation ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Young Adult

Objective]To investigate the change of spinal pro?inflammatory cytokines in a rat model of fentanyl induced acute hyperalgesia.[Methods]64 male SD rats were divided into 2 groups(n=32),fentanyl group and NS group. The rats were subcutaneously injected with fentanyl (60 μg/kg) or normal saline (1.2 mL/kg) 4 times with 15?minute intervals. Mechanical nociceptive thresholds and thermal nociceptive latency were measured via the tail pressure test(Tail flick thresholds,TFT) and paw withdrawal test(Paw withdrawal latency,PWL)on the day before,at 1,2,3,and 4 hour and on 1~5 day after injection. 4 rats were killed concomitantly and the lumber spinal cord were harvested to analysis the expression of NF-κB,PGE2,TNF-α,IL-1β,and IL-6.[Results]There were no significant changes of TFT,PWL and the expression of spinal inflammatory cytokines such as NF-κB, PGE2,IL-1β,and TNF-αcompared to baseline of rats treated with normal saline. The value of TFT ,PWL in fentanyl group raised to the highest(above the baseline)at the 1st hour after fentanyl injections and decreased thereafter,reached the lowest at the 1st day, raised increasinglyand up to baseline on the 3 day after injection. NF-κB,PGE2,IL-1β,and TNF-αincreased at the 4th hour,on 1 and 2 day and IL-6 increased at the 4 hour and onthe 1 day after fentanyl injections.[Conclusion]Subcutaneously injection of fentanyl induced significant mechanical and thermal hyperalgesia and increased spinal pro?inflammatory cytokines parallelly , indicated that fentanyl induced acute hyperalgesia is associated with spinal inflammatory reaction in rats.

The nitrite-reducing activity of aerobic denitrifying bacterial strain N6-1 was studied. It showed that the nitrite-reducing activity reached the highest at 30℃, 120 r/min, pH 8.5 and C/N ratio 12, using CH3COONa and NaNO2 as the sole carbon source and nitrogen source, respectively. When the initial NaNO2 concentration was 2 g/L, NO2--N was reduced completely after 20 hours cultivation with the reducing rate of 20.3 mg/L?h. There would be no effect on its nitrite-reducing activity in the present of 1.5% NaCl or 1% peptone. The cell concentration could reach 1.2?1011 CFU/mL after 24 hours cultivation in 10 L fermentor.

Objective To analyze the related factors associated with the effect of Roux-en-Y gastric bypass in Goto-Kakizaki rats.Methods In GK rats undergoing Roux-en-Y gastric bypass,the weight,food intake,and the fasting blood glucose were retrospectively analyzed before surgery and 1,3,6,12,24 weeks after surgery,and the glycosylated haemoglobin concentrations were retrospectively analyzed before surgery and 4,12,24 weeks after surgery.According to preoperative glycosylated haemoglobin concentrations,GK rats were divided into 3 groups(group A,HbAlc 6.5％-7.9％; group B,HbAlc 8.0％-9.9％ ; group C,HbAlc ＞ 10％).Results The fasting blood glucose and the glycosylated haemoglobin concentrations significantly decreased 1 week after surgery(P ＜ 0.01),on 24 weeks after operation,the fasting blood glucose levels decreased from(12.1 ± 3.0)mmol/L to(7.6 ± 1.3)mmol/L,and the glycosylated haemoglobin concentrations declined from(9.2％ ± 1.8％)to(6.3％ ± 0.8％).Preoperative fasting blood glucose was(11.1 ± 2.2)mmol/L and(15.8 ± 2.3)mmol/L respectively,and the preoperative glycosylated haemoglobin concentration was(8.6％ ± 1.4％)and(11.5％ ± 1.4％)respectively(P ＜ 0.01).Non-conditional Logistic regression analysis identified preoperative glycosylated haemoglobin concentrations as an independent predictor for the cure rate(P ＜ 0.01); Differences were significant between group A and group C,and group B and group C on the efficiency of the surgery(P ＜0.01).Conclusions Significant correlation established between glycosylated haemoglobin concentrations and the efficiency of the surgery,especially for those rats with preoperative HbAlc ＜ 10％.

Objective To investigate the tissue stimulation of copper ions to the smooth muscle of murine terminal rectum.Methods In this study copper needle used for electrochemical therapeutic equipment was punctured into submucosal smooth muscle tissue of terminal rectum above the dentate line in 65 male rats.Rats were then divided into 5 groups (10 rats in each group) at random and sacrificed at 1, 3, 4, 6, 8 weeks after respectively, samples were sent for observation of macroscopic and microscopic tissue reaction.Results Rats had no abnormal histological change in the puncture points grossly. Undor microscope, mild edema was detected in the submucosal layer in 1 week group. Remarkable infiltration of inflammatory cells and lymphocytes subsided 3 weeks after, and disappeared five weeks after.Smooth muscle was normal microscopically in all groups.In contrast, platinum and steel needles caused infiltration of plasma cells and neutrophils, ulcers and small abscess formation in around puncture points.Conclusion Inflammatory reaction was induced after puncturing of copper needle into the tissues and the tissue reaction disappeared after 5 Ws.In contrast with platinum and steel needles, copper needle was non-traumatic to the smooth muscle tissue.

Adult ; Humans ; Male ; Military Personnel ; Models, Biological ; Muscle Contraction ; physiology ; Physical Endurance ; physiology ; Reference Values