Objective To investigate the influence of case environment, extracting position and method on STR typing of nail DNA. Method Decayed body nails were selected from cases. Nails nuclear DNA were extracted from different positions, digestion system and time. After phenol/chloroform extracting and PCR reaction, the STR typing had been studied. Results Nail samples from middle and root positions offer satisfactory results. Conclusion The nail is a useful sample for forensic cases.

Objective To construct the expression vector and express the recombinant human L-selectin in prokaryotic system, and purify the aimed protein for the preparation of rabbit anti-human L-selectin polyclonal antibody. Methods The partial cDNA of human L-selectin was amplified from the open reading frame (ORF) of N-terminus sequence of L-selectin containing 153 amino acids by PCR, then cloned into the prokaryotic expression vector pQE40 at restriction sites BamH Ⅰ and Hind Ⅲ. The recombinant expression plasmid pQE40-L-selectin was transformed into E. coli M15 for expression. The fusion protein including 6-His-tag was purified by Ni-NTA chromatographic column and analysed by SDS-PAGE. The purified protein was used to immune rabbit for preparing polyclonal antibody. Western blotting analysis and dot immunoblot assay (DIBA) were used to test the titer of the antiserum. Results The expression plasmid pQE40-L-selectin was constructed and confirmed with restriction enzyme digestion. The quantity of the purified protein from E. coli M15 was 600 mg ? L-1. By immuning the rabbit, the polyclonal antibody was successfully prepared. The results of dot immunoblot assay (DIBA) showed that the antiserum had the high titer (1 : 1 000). Conclusion The recombinant human L-seleclin prolein can express with high efficiency in E. coli M15. The prepared polyclonal antibody has a high titer.

AIM To establish a dysmenorrhea model in mice. METHODS The mice were given with some kinds of oestrogens once a day for 3～25 days. On the last day, the mice were injected intraperitoneally with oxytocin and the number of twisting body was recorded to evaluate the intensity of dysmenorrhea. The optimum conditions to establish the model was analysed statisticly. RESULTS The optimum oestrogens was stilbestrol. Stibestrol should be given for 12～15 days. The regression equation of the dose effect curve of stibestrol was =0 03?0 04 X, r =0 9688. The optimum dosage of oxytocin was 20 U?kg -1 . The dysmenorrhea model in mice could be preserved for about 7 days. 90% of the twisting body reactions concentrated in 0～30 minutes after oxytocin was given. The effect of oxytocin (20 U?kg -1 ) had significent difference with that of prostaoglantin (1 3 mg?kg -1 ). The test of uterus in vivo showed that stilbestrol could increase the uterine contraction frequency and strengthen the contractility. The dysmenorrhea model in mice was testified by some anti dysmenorrhea drugs. CONCLUSION Compared with the hypodermic implantation in rats, the dysmenorrhea model in mice was simple, reliable, economical and testifiable,etc.

Objective To study the value of 320 slice volume CT venography (CTV)and ultrasound in the diagnosis of lower limb deep vein thrombosis(DVT).Methods 51 patients with DVT confirmed by DSA were analyzed retrospectively,with comparing detection rate by direct method of CTV and ultrasound of the emboli in different parts of lower limb.Results In 5 1 patients,48 cases with DVTs were detected by CTV,including 124 thrombi,and 46 cases by ultrasound,finding 86 thrombi.CT diagnosed 34 pelvic deep vein thrombi,and ultrasound found 10.CT diagnosed 25 tibiofibular vein thrombi,and ultrasound found 5.CT diagnosed 2 femoral deep vein thrombi,and ultrasound found 1 1.Conclusion Direct method of CTV and ultrasound have high clinical value in the diagnosis of deep venous thrombosis,the former is better for the thrombosis in the pelvic deep veins and tibiofibular vein,while the latter is better for the thrombosis in the femoral deep vein.

Objective:To evaluate the correlation of inter-individual variation of cyclosporine dosage and blood concentration and CYP3A4 and CYP3A5 polymorphism in renal transplant recipients. Methods:Two hundred and twenty-one renal transplant recipients treated with cyclosporine were genotyped for CYP3A4 rs4646437C>T and CYP3A5 6986G>A using ligase detection reactions. The effects of genetic polymorphisms of CYP3A4 and CYP3A5 on cyclosporine trough concentration (C0/D) and 2 h post-dose concentra-tion (C2/D) during the period of 6 months, 6-24 months and above 24 months after renal transplant were studied. Results:CYP3A5 6986G>A genotype affected C0/D during the period of 6 months, 6-24 months and beyond 24 months (P<0. 05) with the order of CYP3A5 6986GG>GA>AA. CYP3A4 rs4646437C>T and CYP3A5 6986G>A genotype affected C2/D during the period above 24 months (P<0. 05) with the order of CYP3A4 rs4646437 CC>CT>TT and CYP3A5 6986GG>GA>AA. Conclusion: CYP3A5 6986G>A genotype affects C0/D and C2/D of cyclosporine, and CYP3A4 rs4646437 C>T genotype affects C2/D of cyclosporine. The effects of genotypes are varied in different stages.

BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research.
OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s.
METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by
immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein.
RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.

Objective To assess the efficacy of one-stage posterior median incision via costotransverse joint in treating thoracic spinal tuberculosis.Methods Thirty three patients with tuberculosis of thoracic spine undergoing one-stage posterior thoracic spine debridement,bone grafting fusion and posterior instrumentation from July 2011 to October 2013 were included in the study.There were 18 males and 15 females.The age was from 17 to 39 years old with an average of 29.5.The course was from 5 to 11 months with an average of 7.3.The Cobb angle was from 19° to 42° with an average of 29.8°.There were 5 in upper thoracic spine,17 in middle thoracic spine and 11 in lower thoracic spine.6 were in Frankel scale grade C,11 were in grade D and 16 were in grade E before surgery.Postoperative kyphosis correction,recovery of neurological function and bone fusion were observed.Results All the surgeries were completed successfully.The operation time was from 120 to 230 min with an average of 183.The blood loss during operation was from 480 to 700 ml with an average of 530.All 33 patients were followed-up for 12-36 months with an average of 22.7 months.The Cobb angle was from 8° to 15° with an average of 10.2°,the correction rate was 77％.The Frankel scale of 11 patients recovered from D to E,2 recovered from C to D and 4 recovered from C to E.The postoperative kyphosis correction and Frankel scale were significantly improved,all patients had a 100％ bone fusion rate and there were no internal fixation loosened or shift of graft bone at the last follow-up.ESR and CRP were returned to the normal range and the tuberculosis symptoms disappeared.Conclusions One-stage posterior median incision via costotransverse joint can complete surgery by the same position and the same incision in treating thoracic spinal tuberculosis with safety and good clinical efficacy.

Objective The prognosis of patients with acute myocardial infarction (AMI) is related to age,comorbidities,and other factors,in which non-thyroid sick syndrome (NTIS) may also be an important factor.In this study,determination of blood free triiodothyronine (FT3) was used to explore the short-term and long-term mortality relationship of NTIS with acute myocardial infarction.Methods A total of 1 019 cases of newly diagnosed patients with acute myocardial infarction were enrolled.According to FT3 levels,the enlisted subjects were divided into Quartile 1-4 groups; survival group and non-survival group; normal thyroid function and NTIS group.The enrolled subjects were followed-up for 6-90 months,with the median follow-up time of 44.5 months.Using logistic regression and Cox hazards model,the relationships of short-term and long-term mortality in AMI with NTIS or FT3 were compared.Results The incidence of NTIS in patients with AMI was 27.78％.With the progressively decreasing FT3 and FT3/FT4 ratio,the mortality rates were progressively increased (Quartile 1 group 9.4％,Quartile 2 group 13.8％,Quartile 3 group 14.3％,Quartile 4 group 34.0％,P＜0.01).After being adjusted,FT3 was the strongest influencing factor of mortality within 30 days (RR =0.212,95％ CI 0.125-0.359).Multivariate Cox regression analysis showed that FT3 was independently associated with long-term mortality (RR =0.674,95％ CI 0.514-0.885).Kaplan-Meier showed significant difference in mortality between quartile 1-3 groups and the Quartile 4 group.Even FT3 level was within the low normal range,it was related with the mortality in AMI.Conclusions NTIS is common in patients with AMI.After being adjusted,FT3 was the strongest predictor of mortality within 30 days,and low FT3level in AMI patients was an independent risk factor for long-term all-cause mortality.Even FT3 level was within the normal range,it was still related with mortality in myocardial infarction.

Objective To evaluate and compare C5 palsy and closure of the opened lamina after expansive open-door Laminoplasty (EOLP) with miniplate or suture/anchor fixation.Methods Between January 2011 and January 2013,a total of 142 patients with cervical myelopathy who were treated by EOLP were divided into hinge-side fixation group (fixed with suture/anchor,78 cases)and open-side fixation group (fixed with miniplate,64 cases).The Japanese Orthopaedic Association (JOA) score was used for neurological assessment and recovery rate (RR) counting.Opening angles,cervical curvature index (CCI),posterior shifting of spinal cord (PSSC) and severity of cord compression were recorded and compared.Results All patients in both group were followed up for more than 12 months.All incisions healed by first intention.C5 palsy occurred in 9 patients (9/78,11.5％) of hingeside fixation group,and 1 patients (1/64,1.6％) of open-side fixation group,showing significant difference (P=0.047).Opening angles and PSSC in hinge-side fixation group were greater than that in open-side fixation group.PSSC of 10 patients with C5 palsy were 3.97±1.19 mm,and greater than that of other patients without C5 palsy 2.57± 1.01 mm.There was no significant difference in CCI before (12.23％±3.70％,11.38％±4.29％) and 1 week (12.12％±3.77％,11.31％±4.35％) after operation.No significant difference was found in JOA scores (12.35±1.09,13.55±0.91),JOA improvement rate (64.24％±9.49％,61.78％±11.48％) and cord compression (0.74±0.71,0.75±0.67) at 12 months after operation.In 6 months postoperatively,27％ of patients in hinge-side fixation group,none in open-side fixation group were identified with 10％ decrease or more in opening angles of lamina.Conclusion EOLP with miniplate fixation has the same clinical outcome as fixed with suture/anchor,but will reduce the incidence of C5 palsy and prevent further closure of the opened lamina.

Objective To analyze the quality of life among adults with chronic suppurative otitis media,and to research the chan-ges of the life quality between preoperative and postoperative.Methods We modified the chronic ear survey(CES)through inter-view adults with chronic suppurative otitis media.The modified scale (Chronic Suppurative Otitis Media Outcome Survey)was ad-ministered to 110 patients in a prospective manner,and then was validated according to established criteria for reliability and validi-ty.Then we assessed the outcomes of surgeries for chronic suppurative otitis media.Results The chronic suppurative otitis media outcome survey includes 17 entries,and was divide into four dimensions.Excellent test-retest reliability was obtained for the survey score (R=0.967).Cronbach′s α correlation coefficient were calculated as 0.864 for the total survey.Criterion validity showed a high correlation between scores on chronic suppurative otitis media outcome survey and scores on CES (R=0.977,P <0.01).U-sing principal components extraction with orthogonal rotation,it was performed on the composite data set,and this yielded a four-factor solution that explained 70.394% of the variance.The average score of patients before surgery was 51.660±10.762,post-op-erative scores was 75.893 ± 7.734.The total score wasn′t significantly changed after the surgery,and the average value was 24.23±7.67 (t=24.653,P <0.01).Conclusion The chronic suppurative otitis media outcome survey is a reliable and valid meas-ure of quality of life for adults with chronic suppurative otitis media,and it is more suitable than the CES in outcomes studies and clinical trials.