Objective The aim of this study is investigate the relationship between ACE gene I/D polymorphism and cerebral stroke.Methods 131 patients with cerebral stroke were recruited. The control group includes 50 age-match health cases.The I/D polymorphism of ACE gene was analyzed with PCR amplification.To avoid the mistyping, we used an insertion-specific prime.Results DD genotype shows clear differences among control, infarction and hemorrhage groups(?~2=13.87,P=0.008)。 The frequencies of DD genotype in the patients with cerebral stroke was higher than normal controls(?~2=9.21,P=0.002;?~2=8.76, P=0.003). The D allele also exhibits among control, infarction and hemorrhage groups(?~2=14.23,P=0.005)。 The frequencies of D allele in the patients with cerebral stroke was higher than normal controls(?~ 2 = 11.17,P=0.001;?~2 = 10.87,P= 0.001).Conclusions There was a positive association between the D allele of the ACE polymorphism and cerebral stroke.

Objective To construct the recombinant human metapneumovirus(hMPV) (defined as rhMPV NL/1/00 GFP) in vitro by reverse genetics technique. Methods BSR-T7 cells were transfected using LipofectAMINE 2000 with the full-length cDNA plasmid, and four major protein expressing plasmids, pCITE-N, pCITE-P, pCITE-M2.1 and pCITE-L. After 3 d, cells were subjected to one -80℃ freeze-thaw cycle to prepare lysates. The supernatant of lysate was used to inoculate Vero-E6 cells. After 1-4 d, cells were found for the obvious development of cytopathic effects under light microscope and green fluoroscopic signals under fluorescence microscope, and were observed up to 10 d. The supernatant were collected to de-tect virus titer. Viral RNA was extracted from the supernatant and reverse transcriptase polymerase chain re-action (RT-PCR) was used to amplify N, F and G genes of rescued virus. Results Cytopathic effects and green fluoroscopic signals was readily and obviously observed after 1-4 d post-inoculation in Vero-E6 cells, then cytopathic effects got worse and green fluoroscopic signals became stronger gradually up to 10 d. The ti-ters of the 1st, 5th, 10th,15th and 20th generation virus ranged from 105.0 to 106.5 TCID50/ml. Amplicons with size of 910 bp (N), 450 bp (F) and 980 bp (G) by RT-PCR were accordant with expectant. Nucleotide sequence analysis of above cDNA fragments showed 100% similarity with reported sequence of hMPV NI/1/00 strain. The recombinant virus was genetically constant and GFP-labeled after 20 passages in Vero-E6 cells. Conclusion Recombined hMPV was successfully rescued by reverse genetics technique. This study lays ground for exploring pathogenesis of hMPV infection and development of hMPV attenuated vac-cines.

Objective To evaluate the reliability and accuracy of three-dimensional digital fusion anatomy in the preoperative evaluation and therapeutic strategy choice of intracranial tumors.Methods MRI scan,including regular MRI,MRA,MRV and DTI,were performed in 87 case.Then tumor themselves as well as tumor-associated structures were reconstructed and fused through iPlan 2.6 software.Based on the reconstructed images,therapeutic strategy were established,preoperative and intraoperative imags were compared.Results The digital reconstruction were successfully finished in all cases.Meanwhile,digital images,originally radiological images and actual images matched well.No approach-associated complication were met in our series.Among tumors in the convexity,the relationship of the reflux veins and the tumors were divided into three types:anterior(12 cases),posterior(19 cases) and overriding(3 cases).All of the relationships were seen in the preoperative fusion image,and the veins were all effectively protected during operation.The tumor-associated arteries could be pushed or wraped by the tumors,and the three-dimensional fusion image could provided their virtually aberrant pathway as well as their relationship with tumor.During microsurgical managment of tumors in the deep brain parenchyma,safe approach were found with the help of comprehensive understanding of the tumors and their adjacent structures.Conclusion Three-dimensional digital fusion anatomy can vividly and accurately display full rang of information about the tumor,and facilitate tumor treatment safely.

Objective To establish the plaque assay for the titration of GFP-labeled recombinant human metapneumovims(rhMPV). Methods Vero-E6 cells were selected as host cells for titration. GFP-labeled hMPV was serially diluted and added to each well to infect the cells. The plates were covered with low melting agarose overlay and incubated for different days in incubator. The plates were then observed under fluorescence microscope for plaques with green flourecence, at the same time, the number of plaques was counted by blue plaque-forming. Results Under the low melting agarose overlay, Vero-E6 cells grew well until the CPE caused by hMPV was seen. Clear green flourescence could be observed the first day post infection, much clearer on the third day post infection but showed somewhat fusion between plaques later on.Blue plaques on the fifth day after infection were large and easy to observe. The recombinant GFP-labeled hMPV could replicate up to 1 × 106 PFU in the Vero-E6 cells. Conclusion Plaque assay for titration of recombinant GFP-rhMPV has been sucessfully established. This methodology will offer a solid base for further studies on pathogenesis and vaccine development of this virus.

Objective To evaluate the efficacy and safety of triethanolamine cream in the treatment of skin ulcer. MethodsA multicenter,single-blind,randomized,positive-control study was conducted.One-hundred and twenty patients aging 18-65 years with skin ulcer were randomly classified into the test and control group at a ratio of 2 ∶ 1 to be treated with triethanolamine cream and recombinant bovine basic fibroblast growth factor gel respectively for 4 weeks.The healing rate of ulcer,granulation tissue production rate and epithelialization rate were calculated.Results After the beginning of treatment,the condition of all patients was improved with time.In total,76 out of 80 triethanolamine-treated patients and 38 out of 40 basic fibroblast growth factor gel-treated patients completed the 4-week trial.Significant differences were observed in the healing rate of ulcer,epithelialization rate and granulation tissue production rate between the test and control group (71.05％ vs.34.21％,P =0.0002; 85％ vs.50％,P =0.0001; 66.25％ vs.37.5％,P =0.0035).No adverse events occurred in any of the patients.Conclusions Triethanolamine cream seems superior to recombinant bovine basic fibroblast growth factor gel with regard to the healing rate of ulcer,epithelialization rate and granulation tissue production rate,and may be a promising drug for the treatment of skin ulcer.

Objective To analyze the risk factors associated with periprosthetic femoral fracture following hemiarthroplasty (HA) for displaced femoral neck fracture in aged patients.Methods From January 2013 to June 2016,120 patients over 80 years old were treated by HA for displaced femoral neck fractures.They were 45 males and 75 females,with an average age of 85.2 years (from 80 to 97 years).Their fractures were Garden type Ⅲ (72 cases) and Garden type Ⅳ (48 cases).The time from injury to operation averaged 5.1 days.The patients were divided into a fracture group and a non-fracture group according to the presence or absence of the periprosthetic fracture.The general data of the 2 groups were compared;multivariate logistic regression analyses were done to indentify the influencing factors associated with periprosthetic femoral fracture.Results The 120 patients obtained a mean follow-up of 26.1 months (from 13 to 48 months).Periprosthetic femoral fracture occurred in 11 cases,giving an overall incidence of 9.2％ (11/120).Compared with the non-fracture group,the average age was significantly older,the incidence of past fractures was significantly higher,significantly more types of uncemented stem were used,and American Society of Anesthesiologists (ASA) grading was significantly more severe for the fracture group (P ＜ 0.05).There were no significant differences between the 2 groups concerning the general data (P ＞ 0.05).Multivariate Logistic regression analyses revealed that age [OR =1.268,95％ CI (1.059,1.517),P =0.010] and type ofuncemented stem [OR =0.072,95％ CI (0.008,0.625),P =0.017] were independent risk factors for periprosthetic fracture.Conclusions The incidence of periprosthetic femoral fracture in the elderly patients may be high following HA for femoral neck fractures.Since age and uncemented stem may be independent risk factors for periprosthetic femoral fracture,surgeons should pay enough attention to them in clinic.

Objective: To compare the performance of Miseq and Ion Torrent PGM platforms and library construction method for next-generation sequencing (NGS) technology for formalin-fixed and paraffin-embedded (FFPE) samples. Methods: A total of 204 FFPE cancer samples including 100 non-small cell lung cancers at the First Affiliated Hospital of Zhejiang University, and 104 colorectal cancers at West China Hospital of Sichuan University were retrospectively selected from January 2013 to December 2016. By using the same samples, DNA was extracted, and the same amount of DNA was used for library construction with the same kit, and sequenced on Miseq and Ion Torrent PGM respectively, after passing the quality control. Any discordant mutations between two platforms were validated by amplified refractory mutation system-polymerase chain reaction (ARMS-PCR) method and Sanger sequencing. Results: A total of 204 FFPE samples were included and 197 samples were successfully analyzed by both platforms. The number of reads generated by the samples on Miseq platform sequencing was higher than PGM platform (median 391 634 vs. 298 030, P<0.01). Alignment with human reference genome showed that the mapping rate of Miseq platform was higher than PGM platform (median 100.0% vs. 99.7%, P<0.01). The median sequence depth of samples on Miseq was higher than PGM platform (median 853× vs. 698×, P<0.01). A total of 236 mutations were detected by two platforms, of which 221 were detected on both platforms, with a 93.6% concordance. Miseq platform detected 11 mutations not detected on PGM platform, while PGM platform detected 4 more mutations not detected on Miseq platform. With validation by ARMS-PCR and Sanger sequencing, Miseq platform was more reliable for low-frequency mutations. The main reasons for the discordant mutations between two platforms were that mutation frequency on undetected platform was lower than mutation reporting range (5%) and FFPE samples were stored for a long time. Conclusions Compared with PGM, Miseq platform shows higher sequencing quality in terms of the number of reads, alignment results and coverage depth, and the test results are more reliable. In clinical practice, the appropriate platform should be chosen based on sample size and actual throughput requirements to aid in the molecular characterization of tumors.

This article discusses the design and implementation strategy of Autopsy and Accident Authentication case seminar, with the aim of linking pathology to clinic medicine, theory to practice, and cultivating and improving the thinking ability of analyzing and solving problems for medical students in their medical study. The article investigates the effects and feedback of six rounds of classes, reports three aspects as strengthening the systemic understanding of basic medicine, improving the application and thinking ability, and promoting the thinking of medical humanism, and also discusses the teaching effects, students' feedback, and the significance and existing defects of the seminar.

The residents who had lived for at least 5 years and aged over 20 years old were sampled from urban to rural districts of Jiangsu Province with a stratified cluster sampling technique. B mode ultrasonography and thyroid function determination were carried out in 6 128 persons. The location, diameter, number, boundary, and calcification in thyroid nodules were described by using 7.5 MHz/50 mm transducer of thyroid ultrasonography. TSH was measured by chemiluminescence immunoassay. Free triiodothyronine(FT3)and free thyroxin(FT4)were measured when TSH was abnormal. The crude prevalence of thyroid nodules was 21.12% in total population, 14.55% in male, and 25.24% in female. The standardized prevalence was 15.69%, 11.20%, and 20.40%, respectively. The prevalence was lower in male than in female, and increased with age(P＜0.05). Thyroid nodules in Jiangsu Province were highly prevalent and more attention should be paid to the follow-up, early diagnosis, and treatment.

Objective To investigate the prevalence and epidemiologic characteristics of hypothyroidism among community population in Jiangsu province. Methods The residents who had lived for at least five years and aged more than 20 years old were sampled from six layers in urban and rural districts of Jiangsu province by a stratified cluster sampling technique. Serum was sampled from 7 122 subjects and sTSH was measured by chemiluminescence immunoassay, and FT3 and FT4 were determined in the subjects with abnormal sTSH level. Results (1) The crude prevalences of overt hypothyroidism and subclinical hypothyroidism were 0.66% and 7.53% respectively in total population, with the respective standardized rates of 0.43% and 6.28%. (2)The prevalences of hypothyroidism and subclinical hypothyroidism were significantly higher in females than in males (both P <0.05). (3) The prevalence of subclinical hypothyroidism was significantly increased with advancing age in both female and males (P<0.05). Conclusion Comparing with hypothyroidism, subclinical hypothyroidism shows higher prevalence in Jiangsu province, and more attention should be paid to the follow-up and diagnosis of subclinical hypothyroidism.