Objective To validate the performance of a PMA-based assay for detection of INH-resistant mutations in MTB and investigate the mutation characteristic of INH-resistance.Methods The MTB standard strain H37Rv was from National Tuberculosis Reference Laboratory,1 wild-type strain and 1 katG S315T ACC mutant strain were from Xiamen CDC,7 MTB INH-resistant strains with known INH resistance mutations were from Shenzhen Center for Chronic Disease Control,Henan CDC,No.309 Hospital of PIA and Xiamen CDC.707 MTB clinical isolates were from Xiamen CDC,Xiamen No.1 Hospital and Zhangzhou CDC,126 sputum samples were from Xiamen Tongan CDC.The genomic DNA of the MTB standard strain H37Rv,7 MTB INH-resistant strains and 833 clinical samples,were extracted with Xiamen Zeesan Mycobacterium tuberculosis Isoniazid-resistance Mutation Test Kit using the thermal lysis method.The genomic DNA of 1 wild-type strain and 1 katG S315T ACC mutant strain were extracted with AxyPrep Bacterial Genomic DNA Miniprep Kit.The mutations were discriminated by the △Tm between the samples and wild type control in katG315 codon,inhA promoter -17 to -8 region,ahpC promoter region -44 to -30 and - 15 to 3,and inhA94 codon.A 10-fold dilution series of MTB DNA from 3 × 105 to 300 copies/ reaction obtained from a wild-type strain and a katG S315T ACC mutant strain,respectively,were prepared to determine the analytical sensitivity.Seven MTB INH-resistant strains with 9 predetermined mutations were used for the analytical specificity assay,and 5 mutants of which were used for the repeatability assay.The clinical detection performance of PMA assay were confirmed by the sequencing method in 833 samples.Results Results could be obtained within 3 hours from DNA extraction to PMA assay,including 46 samples in a standard 96-well real-time PCR instrument simultaneously.The analytical sensitivities of PMA were 300 copies/reaction for both the wild-type strain and katG S315T ACC mutant strain.Nine INH-resistant point mutations could be discriminated and 5 of which had standard deviations of melting temperature less than 0.5 ℃.Fully concordant results of mutant locus between PMA assay and sequencing were obtained in all 162 mutant samples.INH-resistant mutations in the four loci were found in 19.4％ (162/833) samples by PMA assay in Xiamen and Zhangzhou.Among the 14 lNH-resistant mutant types detected,katG S315T ( AGC→ACC),inhA promoter - 15C→T and katG S315N (AGC→AAC) accounted for 83.3％ (135/162) of the overall mutations.

Objective:To explore and discuss the effect of Xiaopi Yueyi formula on behavioral changes in rats with chronic fatigue syndrome.Methods:24 rats were divided into group model(16 rats) and control(8 rats) at random,and after produced rats with chronic fatigue syndrome,half in the former were taken orally with Xiaopi Yueyi formula while the latter were taken equally normal saline,moreover,methods of Open-Field、tail suspend time and exhaust swimming time were estimated and evaluated.Results:After succeeding in chronic fatigue manifestation in rats of group model,number of parallel and vertical movement diminished markedly,furthermore,time of tail suspend and exhaust swimming prolonged apparently.Nevertheless,Xiaopi Yueyi formula could ameliorate significantly these parameters in rats(P

Objective To study the inter-rater reliability of Wisconsin Gait Scale (WGS) and Gait Abnormality Rating Scale (GARS) in patients with stroke. Methods 20 hemiplegic patients were required to walk on their comfortable speed and videotaped from frontal, backward and lateral. The video recordings were scored with WGS and GARS by 2 experienced physical therapists. Intraclass correlation coefficient (ICC) was calculated for the scores in each category and the total score. Results ICC for the WGS were 0.372~1, and were 0~0.875 for the GARS. Conclusion WGS is more appropriater to assess the gait of hemiplegic stroke patients than GARS.

Objective To evaluate the potential use of a probe melting analysis (PMA) assay in detecting the embB mutations which confer resistance against ethambutol in Mycobacterium tuberculosis. Methods The analysis sensitivity and specificity of PMA were investigated by detecting a serially diluted H37 Rv DNA and a reference panel from National Institute for the Control of Pharmaceutical and Biological Product. Six hundred and thirteen sputum samples were collected from the Xiamen Center for Disease Control and Prevention, Xiamen First Hospital and Center for Zhangzhou Disease Control and Prevention from September 2009 to April 2010. The PMA assay was then evaluated by detecting 613 clinical isolates and the results were compared with the sequencing results. Results The PMA assay could specifically detect Mycobacterium tuberculosis and had a limit of detection of 3 copies per reaction. The assay results with 613 clinical isolates showed that PMA gave a 100% concordance with sequencing in the 583 qualified samples, among which 34 were mutations at embB 306,23 at embB 378-380, 3 at embB 406 and 3 at embB 497. Conclusions PMA assay is a sensitive and specific method enabling efficient detection of common embB mutations causing ethambutol-resistance. The rapidness of this method together with its reliability would facilitate its use in routine testing.

Objective To evaluate the role of mangled extremity severity score(MESS)in res-ervation and amputation of crush lower limbs in earthquake. Methods There were 122 patients with crush lower limb injuries,with MESS≥8 points in 34 patients who were primarily amputated,M ESS 5-7points in 19 who were principally preserved and MESS＜5 points in 69 who were preserved by means of debridement,external fixators,plast splints and vaeuum sealing drainage technique.Results All pa-tients were survived.with amputation rate of 29.5%. Conclusion MESS is an important reference for evaluation of reservation and amputation of crush limb injuries caused by earthquake.

Objective To investigate the effect of unfractionated heparin (UFH) on the expression of heme oxygenase-1 (HO-1) in intestinal mucosa of mice with sepsis.Methods Thirty-six male C57BL/6J mice were randomly divided into sham group,cecal ligation and puncture (CLP) group and UHF group,n =12 in each group.Model of intestinal injury in sepsis was induced by CLP.In sham group,the mice were exposed without ligation of cecum.In UFH group,the mice were treated intravenously with 8 U of UFH via the tail vein half an hour before the operation and 12 hours after the surgery respectively.Six mice in each group were randomly chosen at 4 hours and 24 hours after operation to collect inferior vena venous blood samples and terminalileum tissues.The serum levels of interleukins (IL-1 β,IL-6),and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA).The serum level of D-lactate was determined by colorimetry.Pathological changes of ileum tissue and Chiu score were observed after hematoxylin eosin (HE) staining.The HO-1 expression was detected immunohistochemically.Results In sham group,no significant changes in the serum levels of IL-1 β,IL-6,TNF-α and D-lactate were observed.Twenty-four hours after the operation,the structure of intestinal mucosa was basically normal without obvious pathology change and no HO-1 positive cells were found.The serum levels of IL-1 β,IL-6,TNF-α,and D-lactate in CLP group were gradually increased,and they were significantly increased as compared with sham group [IL-1 β (ng/L):40.87±2.88 vs.22.60±2.05 at 4 hours,113.73±3.96 vs.22.07±2.74 at 24 hours;IL-6 (ng/L):63.89±3.26 vs.44.89±3.38 at 4 hours,176.56±5.45 vs.45.76±4.02 at 24 hours;TNF-α (ng/L):194.62± 14.13 vs.152.05±6.22 at 4 hours,599.62± 10.20 vs.155.90± 14.18 at 24 hours;D-lactate (mmol/L):0.24± 0.02 vs.0.19 ± 0.01 at 4 hours,0.33 ± 0.04 vs.0.20 ± 0.02 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,edema and inflammation in ileal mucosa,intestinal villi structural damage were observed,the Chiu score was significantly higher than those in the sham group [4.5 (3.0-5.0) vs.0 (0-1.0),P ＜ 0.05],and a small amount of HO-1 positive cells were localized in the intestinal mucosa.Compared with CLP group,the serum levels of IL-1 β,IL-6,TNF-α,and D-lactate of UFH group were significantly decreased [IL-1 β (ng/L):31.53 ± 2.90 vs.40.87 ± 2.88 at 4 hours,61.13 ± 2.80 vs.113.73 ± 3.96 at 24 hours;IL-6 (ng/L):51.16 ± 5.68 vs.63.89 ± 3.26 at 4 hours,81.16 ± 4.54 vs.176.56 ± 5.45 at 24 hours;TNF-α (ng/L):171.76± 5.60 vs.194.62± 14.13 at 4 hours,328.48 ± 10.79 vs.599.62± 10.20 at 24 hours;D-lactate (mmol/L):0.21 ±0.01 vs.0.24±0.02 at 4 hours,0.24±0.02 vs.0.33±0.04 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,intestinal injury was ameliorated,the Chiu score was significantly lower [1.5 (1.0-5.0) vs.4.5 (3.0-5.0),P ＜ 0.05],and HO-1 positive cells in the intestinal mucosa was remarkably increased.Conclusion UFH can enhance the expression of HO-1 in intestinal mucosa,reduce the release of inflammatory factors,ameliorate the intestinal inflammatory response,and thus play a protective role in intestinal tissue in mice with sepsis.

Objective To investigate the optimal time for double balloon enteroscopy (DBE) in patients with obscure gastrointestinal bleeding (OGIB).Methods Data of 78 patients with OGIB who underwent DBE from January 2009 to November 2013 were retrospectively analyzed.They were classified into two groups:emergency DBE and non-emergency DBE.The demographic and clinical features and outcomes of DBE,the time of examination and complications were compared.Results The overall diagnostic yield of DBE was 48 lesions (61.54 ％).The overall diagnostic yield of emergency DBE group was 77.14％,which was significantly higher than that in non-emergency DBE group (48.83％) (P =0.019).The time of examination in emergency group was shorter than that of non-emergency group with significant difference (P =0.031).Conclusion Emergency DBE takes less time and yields a higher rate of detection.Patients withOGIB should receive DBE as soon as possible.

Objective To find the plantar pressure readings which best indicate foot inversion during the stance phase of walking among hemiplegic stroke survivors.Methods Twenty-two hemiplegic stroke survivors who were able to walk without extra aid were recruited as the experimental group, while 17 healthy elderly men of similar age and body weight were selected as the control group.Those in both groups were asked to walk at their preferred speed over a Footscan device which measured medio-lateral pressure ratios, maximum plantar pressures and the contact areas of both feet.The Clinical Spasticity Index (CSI) was used to evaluate the affected feet.Results The average medial forefoot pressure of the affected side in the experimental group was significantly lower than that of the control group, but their average mesopodium and heel pressure was significantly higher.The average pressure applied by the great toe on the uninjured side in the experimental group was significantly smaller than that of the control group, but the average mesopodium and heel pressure of both feet among the hemiplegics were significantly higher than those of the control group.Among the experimental group, the average medial forefoot pressure of the affected foot was significantly greater than that of the healthy foot.The average contact area of the great toe on the affected side was significantly bigger than was observed in the control group.That of the medial forefoot was, however, significantly smaller than in the control group.There was no significant difference in the contact area between the healthy and affected feet in the experimental group, though the maxmium medio-lateral pressure ratios of their full feet and forefeet on the affected side were significantly lower than those in the healthy group.No significant differences in the maxmium medio-lateral pressure ratios of the heel were observed between the two groups, nor of the full feet, forefeet and heels of the affected and unaffected sides in the experimental group.The patients demonstrated consistently reduced joint mobility on both sides during the stance phase, coinciding with increased inversion.A significant negative correlation was found between the maxmium medio-lateral pressure ratios of the full foot and the maximum pressure of the lateral part of forefoot in the experimental group, but there was no significant correlation with contact area or CSI.Conclusions Plantar pressure data can be used to describe the amount of foot inversion in the stance phase of walking with hemiplegic patients after stroke.The maxmium medio-lateral pressure ratios can effectively reflect their foot inversion.

A new dibenzocyclooctadiene lignan, renchangianin E (1) was isolated from the stems of Kadsura renchangiana. Its structure and stereochemistry were elucidated by spectroscopic methods, including 2D-NMR techniques.

Objective To establish an in vitro accelerated release method of triptorelin acetate microspheres with good in vi-tro/in vivo correlation(IVIVC). Methods The in vivo release of triptorelin acetate from microspheres was obtained by residual method in rats. Influences of pH value,concentration of ethanol,temperature,rotation speed and concentration of antiseptic on the in vitro accel-erated release were studied,then the correlation between in vitro accelerated release and in vivo release of the microspheres was estab-lished by adjusting the release conditions. Results The in vitro accelerated release medium of triptorelin acetate microspheres composed of 15%ethanol solution(containing 0.06%Tween 80 and 0.1%benzalkonium chloride)at 55℃with rotating rate of 200 r/min. The cumulative release of triptorelin acetate from microspheres was 87.35%at 30 h under accelerated release condition,equivalent to in vivo release for 30 days. The established in vitro accelerated release had a good correlation with that of in vivo(y=0.8845x+12.4510, R2=0.9938). Conclusion The in vitro accelerated release of triptorelin acetate microspheres could correlate well with in vivo release and has a potential application in rapid and effective evaluation of triptorelin acetate microspheres.