Objective To express, purify and refold recombinant luteinizing hormone releasing hormone-angiogenin(LHRH-Ang) toxin using E. coli. expression system. Methods Recombinant LHRHAng expression vector was constructed by replacing of EGF fragment in plasmid pET28a/EGF-Ang with LHRH-P Ⅱ fragment amplified from plasmid pET28/MSH-PEA0. DNA sequencing would be used to verify the correction of fused LHRH-P Ⅱ -Ang gene. Then, E. coli strain BI21 (DE3) was transformed by pET28a/LHRH-Ang vector. Expression of recombinant LHRH-Ang toxin was induced by Isopropyl-β-D-Thiogalactoside( IPTG ). Refolding effects of gradient dialysis was evaluated by SDS-PAGE. Results Prokaryotic expression vector pET28a/LHRH-Ang, containing LHRH-P Ⅱ -Ang fusion gene, was constructed by PCR amplification, restriction enzyme digestion and ligation method. Sequence correction of fusion gene was confirmed by DNA sequencing After IPGT induction, recombinant LHRH-Ang protein was expressed in BL21 ( DE3 ) as inclusion body ,it took 18.43% of total protein. Inclusion body was resolved in 8 mol/L urea and purified by DEAE-Sepharose FF column, the purity was 85%. Recombinant LHRH-Ang toxin was refolded and concentrated by gradient dialysis and PEG 20000, respectively. Conclusions Recombinant LHRH-Ang protein was expressed in E. coli and refolded successfully.

Objective To explore the etiology,clinical presentation and treatment of patients with spontaneous spinal epidural hematoma (SSEH). Methods Twelve patients diagnosed as having SSEH, admitted to our hospital from January 1995 to December 2010, were chosen in our study; 10 underwent surgical operation and 2 chose conversational therapy; the characteristics of onset,clinical presentations, diagnosis, treatment and results of follow-up were analyzed. Results No obvious reasons induced the onset of SSEH in these patients and all manifested as loss/decrement of myodynamia of the involved limbs,and bowel and bladder dysfunctions.Follow-up was performed for 6-18 months; 2 patients received hematoma lesion dissection by spinal canal decompression enjoyed entire recovery of neural dysfunction, while 8 showed no significant amelioration in spinal cord function besides mild improvement of bowel and bladder function; in the 2 patients chosing conversational therapy,1 achieved total recover of the nerve funtion and the other one partially recovered. Conclusion The etiology of SSEH remains indefinite and the prognosis is usually disappointed; permanent spinal cord dysfunction is seemly inevitable in most cases; prompt surgical intervention are of great importance for prognosis of SSEH and should be the first choice of therapy.

Objective To report a new approach,endoscopic transoral approach for the resection of jugular foramen schwannoma.Methods Nine patients with jugular foramen schwannoma ( three males and six females,ranging in age from 15 to 61 years old ) were treated by direct surgery via a pure endoscopic transoral approach to the jugular foramen. Eight patients complained of hypoglossal nerve palsy with hemiatrophy of the tongue; six cases complained of vagus nerve palsy. Three cases complained of glossopharyngeal nerve palsy,one case complained of facial nerve palsy and hearing loss.Results The nerves in this area were preserved and radical intracapsular removal of the tumor was performed via endoscopic transoral approach in the nine cases.Tumor removal,as assessed by intraoperative endoscopic inspection,postoperative magnetic resonance imaging and clinical evaluation,revealed all tumors were completely removed.One patient suffered from temporary swallowing difficulties and temporary right vagus palsy Ⅰ day after surgery.There were no others intraoperative and postoperative complications.All patients were followed up for 4 -29 months,no recurrences were occured in all these patients and the muscle bulk,motor and the pre-postoperative swallowing fuction,the vagus palsy,the facial nerve palsy and hearing loss had improved in these patients.Conclusion The endoscopic transoral approach and intracapsular removal of the tumor provided for successful minimally invasive surgery in the jugular foramen schwannomas.

Objective To explore the ability and mechanism of insulin-like growth factor l (IGF-1) induced bone mesenchymal stem cells ( BMSCs) differentiation into cardiomyocyte-like cells ( CLCs).Methods BMSCs were isolated and purified in vitro.BMSCs were treated with control medium and 15 ng/ml IGF-I for 3,7, 14 and 21 d, respectively. The expression of Troponin-T ( TNT), Troponin-I (TNI) and pIGF-1R were c~etected by immunocytochemistry and Westem blot.In another experimental setting,BMSCs were treated with control medium and 15 ng/ml IGF-l,ICF-1 antaS;onist I-Ome AG538 (300 nmol/L) and 300 nmol/L I-Ome AG538 + 15 ng/ml IGF-1 for 3 t0 48 h,respectively.Phosphorylation status of ERKl/2 and AKT,the two downstream mediators of mitogen-activated protein kinase ( MAPK) kinase and phosphatidylinositol 3-kinase ( PBK) pathways,were detected by immunocytochemistry and Yvestem blot.Results After 3 t0 21 d exposrue to IGF-1, the expression of pIGF1R,TNT and TNI were significandy higher in 'IGF-1 group than those in control group,pIGF-IR peaked 14 d (all P ＜0.05).After 3 and 6 h treatment,the ratio of pAKT/AKT(0.17 ± 0.03)and pERK1/2/ERK1/2(0.06 ± 0.03)were significantly downregulated in I-Ome AG538 group compared to control group (1.00 ±0.05) (all P ＜ 0.05).The ratio of pAKT/AKT(1.00 ± 0.07) and pERK1/2/ERK1/2 (1.00 ± 0.09) were significantly upregulated in IGF-1 group compared to control group (0.72 ± 0.05) (all P ＜ 0.05),but the ratio of pAKT/AKT(0.31 ± 0.10)and pERK1/2/ERK1/2 (0.39 ±0.04) were significantly downregulated in I-Ome AG538 group compared to control group (0.63 ± 0.05) (all P ＜ 0.05),the value of gray scale of TNT (195.06 ± 5.98) and TNI (198.32 ± 3.46) in I-Ome AG538 + IGF-1 group were significantly upregnlated than that in IGF-1 group for TNT (188.70 ± 5.35) and TNI (176.10 ± 4.96) (all P ＜ 0.05).Conclusions IGF-1 could induce BMSCs differentiation into CLCs in vitro by activating MAPK and PI3K signaling pathways.

Purpose To investigate the consistency and clinicopathologic correlation of BRAFV600E protein expression and gene mutation in papillary thyroid carcinoma. Methods BRAFV600E protein expression and genn mutation was detected respectively by immunohistochemistry of SP and real time-PCR, then the consistency between the both methods was analyzed by Kappa-test, the correlation between BRAFV600E and clinicopatho-logic parameters was analyzed by Chi-square test in papillary thyroid carcinoma. Results The gene mutation and protein expression rates of BRAFV600E were 89.3％ and 88.3％, respec-tively, the differences were not significant, the concordance rate of the both methods was 97.0％, Kappa value was 0.847, the consistence was higher, meanwhile the mutation rates between age ＜45 and ≥45 were respectively 96.8％ and 85.9％, there were significant differences, the positive rates of the both detec-tion methods were higher in thyroid capsule invaded group than non-invaded group, the differences were significant. Conclusion The both methods have higher consistency, the immunohisto-chemistry can be used as an initial screening tool for detecting gene mutation, the gene mutation of BRAFV600E is significantly associated with age and capsule invasion, the relationship is not found between BRAFV600E mutation and the other clinicopatholog-ic parameters.

Objective Dyslipidemia and chronic inflammation are risk factors of cardiac fibrosis.This study was aimed to investigate their possible synergetic effects and underlying mechanisms on progression of cardiac fibrosis in apolipoprotein E knockout (ApoE-/-) mice.Methods Twenty-four ApoE-/-mice were divided into normal chow diet (control),high fat diet (HFD group),and HFD plus subcutaneously injection of 10％ casein (inflammation group) for 8 weeks.Lipid profile and serum amyloid A (SAA) were examined by clinical biochemical assays and Enzyme-Linked Immunosorbent Assay,respectively.Hematoxylin-eosin staining (HE) and Masson staining were used to evaluate the myocardial accumulation of lipid and collagen.Collagen Ⅰ protein expression was detected by immunohistochemical staining.Endothelial-to-mesenchymal transition related protein expressions were determined by Western blot.Results Serum SAA level was significantly higher in inflammation group [(127.42 ± 26.99) ng/ml] than in control [(15.40 ± 7.62) ng/ml] and HFD [(8.17 ± 0.72) ng/ml] group (all P ＜ 0.01).However serum levels of triglyceride,total cholesterol,and low density lipoprotein (LDL) cholesterol were significantly higher in HFD group than in inflammation and control groups[TG (7.53 ± 2.05) mmol/L vs.(3.43 ± 0.79) mmol/L ; TC (27.80 ± 3.99) mmoL/L vs.(14.94 ± 1.92) mmol/L ; LDL-C (11.56 ±2.56) mmol/L vs.(9.46 ± 1.31) mmol/L,all P ＜ 0.05).Foam cell formation in cardiac vessels.myocardial collagen deposit,protein expressions of collagen Ⅰ,CD31,and alpha-smooth muscle actin (α-SMA) were all significantly higher in inflammation group than in HFD group (all P ＜ 0.05) suggesting that inflammation contributes to the phenotype endothelial-to-mesenchymal transition in heart.Conclusion Inflammation exacerbates dyslipidemia mediated cardiac fibrosis in ApoE-/-mice partly through enhancing myocardial endothelial-to-mesenchymal transition.

Objective To identify the impact of age and gender on cardiac structure and left ventricular function in normal Chinese by echocardiography. Methods Cardiac structure, valve flow velocity and cardiac function were measured by echocardiography in 15 692 healthy volunteers. Subjects were grouped by age at 5 years interval in population older than 5 years. Children under 5 years were divided into 3 age groups(<1 years,1-3 years,4-5 years). Hierarchical cluster analyses were performed for ages, based on indexes of cardiac structure and function respectively. Results Six groups (< 1 years, 1-3 years,4-5 years, 6-10 years, 11-20 years, ≥21 years) were generated after the age hierarchical cluster analyses based on index of cardiac structure. Four groups (≤30 years, 31-50 years, 51-80 years,≥81 years) were generated based on spectral current flow. Six groups (< 1 years,1-3 years,4-5 years, 6-10 years, 11-15 years, ≥16 years) were generated baaed on left ventricular systolic function and five groups (≤15 years, 16-30 years, 31-50 years, 51-80 years, ≥81 years) were generated based on left ventricular diastolic function. Cardiac structure index were similar between male and female in age groups ≤ 10 years and significantly lower in females than males in age groups ≥ 11 years (P < 0.05). Valve flow velocity was similar between male and female in various age groups (P >0.05). Left ventricular systolic function was similar between male and female in age groups ≤10 years but was significantly higher in males than females in age groups ≥11 years(all P <0.05). Left ventricular diastolic function was similar between female and male in various age groups (P > 0.05) and equally decreased with aging in both female and male subjects. Conclusions The cardiac development in Chinese population can be divided in 6 phases and becomes stable in subjects older than 21 years, left ventricular systolic function becomes stable in subjects older than 16 years and the left ventricular diastolic function declines physiologically with aging.

Objective To investigate the feasibility and efficacy of tubed pectoralis major myocutaneous flap in the reconstruction of circumferential defects following resection for locally advanced hypopharygeal and cervical esophageal carcinoma. Methods From Dec. 2004 to Oct 2008, 30 patients underwent immediate reconstruction by tubed pectoralis major myocutaneous flap for circumferential defects following resection of primary tumours. Among them, 22 were hypopharyngeal carcinoma, 7 were cervical esophageal carcinoma and one was recurrent laryngeal carcinoma involved the hypopharyngeal lumen. Five of 30 patients had received previous radiotherapy and three had failed in the previous surgical procedure. In this series, 12 patients had total pharyngolaryngectomy and 18 had total pharyngolaryngectomy and partial cervical esophagectomy. Results Postoperative pharyngocutanous fistula formation occurred in 4 patients, 2 of them with previous radiotherapy and 2 with diabetes, and the fistulae healed later. Two patients developed anastomotic strictures at the upper junction, but they had good respondses to dilatation treatment and had satisfactory oral intake. The postoperative follow-up time ranged from 8 to 56 months. Median follow-up was 18 months. One-year survival rate was 71.4% and three-year survival rate was 42.5%. Conclusions The tubed pectoralis major myocutaneous flap is a reliable procedure to reconstruct hypopharyngeal circumferential defects following resection of advanced hypopharygeal and cervical esophageal carcinoma. This method may be the optimal choice for the reconstruction of hypopharyngeal circumferential defects following resection of recurrent carcinoma. The incidence of fistula and stenosis could be kept at an acceptable level.

AIM To establish the quality standard for Weigela japonica Thunb.var.sinica (Rehd.) Bailey (W.j.).METHODS TLC was adopted in this medicinal material's qualitative identification after morphological identification and microscopic identification.The contents of water,total ash,acid-insoluble ash and extracts were detected according to Chinese Pharmacopoeia methods.Then the contents of scopoletin and total coumarins were determined by HPLC and UV,respectively.RESULTS The morphologies and microscopic characters of W.j.could be distinguished from other same generic plants.The clear TLC spot displayed good resolution.The contents of water,total ash,acid-insoluble ash,water-soluble extract and acid-soluble extract were no more than 12.0％,no more than 2.0％,no more than 0.5％,no less than 5.0％ and no less than 4.5％,respectively.Scopoletin and total coumarins showed good linear relationships within the ranges of 1.25-40.0 μg/mL(r =0.999 7)and 2.0-64.0 μg/mL (r =0.999 9),whose average recoveries were 98.19％ (RSD =0.90％) and 99.21％ (RSD =2.5％),respectively.CONCLUSION This accurate and reliable method can be used for the quality control of W.j..

Objective To analyze the status of quality indicators(QI) on specimen acceptability and establish preliminary qual ity specification.Methods Web based External Quality Assessment system was used to collect data of laboratories partici pated in "Medical quality control indicators in clinical laboratory" from 2015 to 2017,including once in 2015 and 2017 and twice in 2016.Rate and sigma scales were used to evaluate incorrect sample type,incorrect sample container,incorrect fill level and anticoagulant sample clotted.The 25th percentile (P25) and 75th percentile (P75) of the distribution of each QI were employed to establish the high,medium and low specification.Results 5 346,7 593,5 950 and 6 874 laboratories sub mitted the survey results respectively.The P50 of biochemistry (except incorrect fill level),immunology and microbiology reach to 6σ.The P50 of clinical laboratory is 4 to 6σ except for incorrect sample container.There is no significant change of the continuous survey results.Based on results in 2017 to establish the quality specification,the P25 and P75 of the four QIs is 0 and 0.084 4 ％,0 and 0.047 6 ％,0 and 0.114 2 ％,0 and 0.078 4 ％,respectively.Conclusion According to the results of the survey,most laboratories had a faire performance in biochemistry,immunology and microbiology,and clinical laboratory needs to be strengthened.Laboratories should strengthen the laboratory information system construction to ensure the actual and reliable data collection,and make a long time monitoring to achieve a better quality.