Objective:To construct the extracellular region of the human TRAIL cDNA expression vector and express and purify the extracellular region of the TRAIL protein. Methods: The mRNA of TRAIL was extracted from CD3 activated normal human PBMC and used as a template for reverse transcription. After PCR amplification, a 730 bp fragment including extracellular region was obtained and cloned into pGEX-2T.The recombinant vector was named pGEX/TRAILex. The pGEX/TRAILex vector was transformed into E.coli DH5a. After IPTIG induced at lower temperature, the collection of the sonicated extract was purified by using the GST agarose 4B. The purified fusion protein was identified by Western blotting with anti-TRAIL McAb.Results:The pGEX/TRAILex was constructed. After IPTG induced,a high level expression of the extracellular region of the TRAIL protein was obtained, SDS-PAGE analysis showed that the recombinant E. coli could express a 54 kD GST fusion protein which accounted for about 28% of the total cellular protein. The study of solubility of expression protein indicated that GST-Tex was expressed predominantly in the soluble form.The purified production was obtained 2.2 mg/L of culture media and the purity of the GST-Tex was more than 95%. GST/TRATLex protein could be recognized by anti-TRAL McAb in Western blot. Conclusion:The expression of recombinant extracellular domain of the human TRAIL protein may be useful for the study of biological functions of TRAIL and it's biotheraphy in tumor.

OBJECTIVE:To optimize base material compositions and preparation technology for compound ciprofloxacin gel.METHODS:4factors,ciprofloxacin,carbomer-940,glycerine,triethanolamine(in formula dosages)were selected as variable factors,the stability of the preparation was taken as the index for investigation and table L 9 (3 4 )was used to conduct the orthogonal experiment.RESULTS:The optimum base material composition and technology were the following,ciprofloxacin0.3%,carbomer-9401%,glycerine7.5%,triethanolamine2%.CONCLUSION:The compound ciprofloxacin gel prepared based on the optimized technology is in conformity with the specification stated in Chinese Pharmacopoeia(2000).

Objective To evaluate the incidence of postoperative epididymitis by laparoscopic cluster ligation and laparoscopic internal spermatic vein high ligation in the treatment of varicocele. Methods Forty-four cases of bilateral varicocele were retrospectively analyzed , who had undergone laparoscopic spermatic vein ligation. The 22 cases were treated with laparoscopic cluster ligation, and the other 22 cases with laparoscopic internal spermatic vein high ligation. The effects and the reasons of two kinds of surgical procedures on postoperative epididymitis were investigated. Results A total of seven patients were affected with postoperative epididymitis. The incidence of laparoscopic cluster ligation was 27.27%. The incidence of laparoscopic internal spermatic vein high ligation was 4.55%. All cases were affected within 4 weeks after surgery. Conclusion It is important to reserve the spermatic artery for decreasing the incidence of postoperative epididymitis.

Objective To analyze the initial response rate of radiotherapy and chemotherapy,and the influence of different treatment modalities on pmgnnsis of primary non-Hodgkin lymphoma(NHL)of the nascl cavity.Methods From January 1993 to December 2002,70 patients were confirmed by histopathology as NHL,including 52 T cell origin,2 B cell and 16 NK cell.The main radiotherapy portal was set in front of the nose with a spade like protrusion,supplement with a portal next to the ear on one side or both sides.Combined portal in facial cervical area was first used when there was invasion of the oropharynx.The median dose to the nasal cavity wag 54.0 Gy with a range of 336 to 66 Gy,given before or during after radiotherapy or alone,consisted of 2-6 cycles chemotherapy of COP,CHOP or COBDP.Prognostic factors were analyzed with Cox model. Results The local control rates was 12.5%, 66.7% and 74.0% in chemotherapy alone,radiotherapy alone,and chemotherapy plus radiotherapy.The 5-year survival rate Wag 12.5%,50.0% and 62.0%,respectively(P＜0.05).Multifaetor analysis showed that tumor extension and treatment methods were the most important prognostic factors besides Ann-Arbor stage,but gender,pathology,age and symptoms had little effect on prognosis.Conclusion Chemotherapy plus radiotherapy group achieved a better survival rate than radiotherapy alone.For some NHL patients with good financial condition,APBSCT is a good choice.

Objective: To compare the difference in somatic gene mutation of PTC subtypes between 114 patients with papillary thyroid carcinoma (PTC) and The Cancer Genome Atlas (TCGA) database. Methods: Totally 114 PTC patients admitted to The First Affiliated Hospital of Nanjing Medical University were recruited. The 18 hotspot genes associated with thyroid cancer were detected in thyroidectomy specimens were using next generation sequencing. PTC data were downloaded from the TCGA database in the cBioPortal website, and the difference in the somatic gene mutation was compared between 114 PTC patients and the TCGA database Results: The 114 PTC patients included 73 women (64.04%) and had a mean age of (39.23±13.18) years. The prevalence of BRAF V600E (66.67% vs. 48.68%), TERTp (3.51% vs. 0.41%), PDGFRA (1.75% vs. 0%), PTEN (3.51% vs. 0.41%) and TP53 gene mutations (4.39% vs. 0.61%) was significantly higher among the 114 PCT patients than in the TCGA database (P<0.05). The prevalence of BRAF V600E (80.88% vs. 54.99%), TP53 (7.35% vs. 0.57%) and TSHR gene mutations (2.94% vs. 0%) was significantly higher in classical PTC(CPTC) patients than in the TCGA database, and the prevalence of BRAF V600E (36.84% vs.13.86%) and TERTp gene mutations (10.53% vs. 0%) was significantly higher in follicular variant PTC (FVPTC) patients than in the TCGA database. According to the American Thyroid Association Risk Stratification of Thyroid Cancer Recurrence, the prevalence of BRAF V600E and TP53 gene mutations was 77.14% and 8.57% among moderate-risk CPTC patients, the prevalence of BRAF V600E gene mutation was 27.27% among low-risk FVPTC patients, and the prevalence of TERTp gene mutation was 33.33% among moderate-risk FVPTC patients, which were all higher than in the TCGA database (55.10%, 0%, 3.28%, and 0%, respectively; P<0.05). Conclusion There are significant differences in the type and rate of somatic gene mutations between 114 PTC patients and the TCGA database.

Objective To study DNA quantification and STR typing of samples pre-treated with pyrami-don. Methods The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accor-dance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24 h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. Results In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Conclusion Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extrac-tion is the best method for STR profiling and DNA extraction.

Objective To investigate the clinicopathological characteristics and postoperative prognosis analysis of intra-thoracic localized Castleman disease (LCD).Methods The clinical data of 9 patients with intra-thoracic LCD who accepted surgical treatment were retrospectively analyzed.There were 5 males and 4 females,with age of (32.8 ± 10.9) years.Two patients complained of chest pain,1 patient suffered from paraneoplastic pemphigus,and the rest were diagnosed by physical examination.Four cases were diagnosed with LCD by preoperative CT examination.Results All patients underwent surgical resection.Four patients were performed open surgery and 5 patients had video assisted thoracic surgery.All patients accepted radical surgery.But 2 of these patients had postoperative complications.One patient was the injury of phrenic nerve and another was pericardial effusion.Patho-histological showed hyaline vascular type of Catleman disease in all patients.All patients survived without recurrence during the follow-up for 2-53 months.Conclusions Intra-thoracic is rare and liable to misdiagnosed.For increasing the preoperative diagnosis rate of LCD,the combined application of imaging tests is important,and clinicians and radiologists should also enhance the awareness of this disease.Complete surgical resection of the tumor is the best therapeutic alternative for intra-thoracic LCD.

OBJECTIVETo analyze the characteristics of autopsies in medical dispute.

METHODSThe data of 202 autopsy cases in medical disputes performed by the Department of Pathology of Jiangxi Medical College from January 2001 to December 2010 were retrospectively analyzed.

RESULTSThe number of autopsy in medical disputes increased year by year. Neonatal, infant and 30 ≊ 60y were more common age groups in dispute; the gender ratio of male to female was 2:1 and more cases were from hospitals in rural areas. Most medical disputes came from in departments of pediatric, obstetrics and gynecology and general surgery. Death caused by cardiovascular diseases ranked at the first place.

CONCLUSIONAutopsy is important in medical disputes to define the cause of death and to preserve evidence.

Adolescent ; Adult ; Aged ; Autopsy ; statistics & numerical data ; Cardiovascular Diseases ; mortality ; Cause of Death ; Child ; Child, Preschool ; Dissent and Disputes ; Female ; Hospital Departments ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Male ; Malpractice ; Middle Aged ; Retrospective Studies ; Sex Ratio ; Young Adult

Objective To explore the anatomical features and the dissection technique of thyrothymic ligament (TTL),and to explore the clinical significance of protecting the inferior parathyroid gland (IPTG) with this structure.Methods Patients who received the initial thyroid surgery in the Department of Thyroid Surgery of the First Affiliated Hospital with Nanjing Medical University from May.2017 to Dec.2017 were prospectively analyzed.We dissected TTL,identified and located the IPTG,described the structural features of TTL,and investigated the position relationship of TTL and IPTG to evaluate the possibility and value of protecting IPTG in situ.Results About 121 patients underwent the dissection,totally 194 sides dissected that included 96 left sides and 98 right sides.TTL was found in 143 sides (73.7％),78 left sides (81.3％) and 65 right sides (66.3％).Nearly 70.6％ IPTG can be proactively identify and located by the TTL during the operation.TTL was a kind of adipose connective tissue that was wide at the bottom and narrow at the top,accompanying with the inferior thyroid vein,from the thymus to the thyroid.76.2％ TTL were attached to the lower pole and the lower 1/3 dorsal of thyroid,containing fat and vessels.33.5％ IPTGs were located in the area surrounding around the ends of the TTL.25.3％ IPTGs were located in the TTL.4.6％ IPTGs were located in the thymus and 7.2％ IPTGs surrounding around the TTL.The incidence rate of post-operation hypoparathyroidism was 14.9％.Conclusions TTL commonly exists and has significant relationship with IPTG.TTL connects thymus and IPTG,which would be considered a complex (thymus-thyrothymic ligament-IPTG complex,TLIC).The meticulous TTL dissection technique will help proactively identify,locate and protect IPTG during operation,and reduce the incidence rate of post-operation hypoparathyroidism.

Objective To investigate the expression of melanoma antigen- encoding gene (MAGE) A1 protein in esophageal squamous cell carcinoma, and explore its correlation with the clinicopathological factors and prognosis. Methods A retrospective analysis was performed on 197 patients with esophageal squamous cell carcinoma who accepted radical surgical treatment from January 2006 to December 2012. The expressions of MAGEA1 protein in these specimens of cancer tissue and cancer adjacent tissue were detected by immunohistochemistry with tissue microarray technology. Results MAGEA1 protein was expressed in cytoplasm and nucleus of tumor cells. The positive expression rate of MAGEA1 protein in cancer tissue was significantly higher than that in cancer adjacent tissue: 73.6％ (145/197) vs. 5.6％ (11/197), and there was statistical difference (P<0.01). The positive expression of MAGEA1 protein had no correlations with sex, age, history of smoking/drinking, family history of upper gastrointestinal cancer, depth of tumor invasion, lymph node metastasis, tumor differentiation, location and TNM stage (P>0.05). Kaplan-Meier survival analysis result showed that the 5-year survival rate in patients with MAGEA1 protein positive expression was significantly lower than that in patients with MAGEA1 protein negative expression (37.2％ vs. 53.8％), and there was statistical difference (P=0.018). Multivariate analysis result showed that MAGEA1 protein positive expression was an independent predictor of prognosis in esophageal squamous cell carcinoma patients (HR=1.91, 95％CI 1.22 to 2.98, P = 0.004). Conclusions The expression of MAGEA1 protein is abundant in esophageal squamous cell carcinoma, and is related to worse clinical outcome. MAGEA1 protein could be a candidate target for tumor immunotherapy.