Objective To establish a method for detecting the EGFR gene mutations by the real-time fluorescence quantification PCR combined with Sanger sequencing and to preliminarily explore its clinical application value .Methods With EGFR gene hotspot mutations region exon 19 and 21 as the research locus ,the specific amplification and the sequencing primer were designed ,the known wild-type and mutant samples were utilized to construct the corresponding plasmid as the standard substance by the TA clone technique .Then the EGFR gene mutation detection method by the real-time fluorescence quantification PCR combined with Sanger sequencing was established and the methodological and the application evaluation were performed .Results The wild-type and mutant standard plasmids of the EGFR gene exon 19 and 21 were constructed successfully .The EGFR gene mutations detection method of the real-time fluorescence quantification PCR combined with Sanger sequencing was established ,which had high sensitivi-ty(101copies/μL)andgoodrepeatability(intra-assayCVandinter-assayCVofthereal-timefluorescencequantificationPCRofex-on 19 and 21 were 1 .42% /3 .52% and 0 .97% /2 .44% ,respectively ) .20 clinical samples were simultaneously detected by this method and the traditional Sanger sequencing ,the results were completely consistent .Conclusion The EGFR gene mutations detec-tion method of the real-time fluorescence quantification PCR combined with Sanger sequencing is successfully established ,which can be used in the clinical sample detection .

Objective To determine the independent risk factors for postoperative nausea and vomiting (PONV) after endoscopic retrograde cholangio-pancreatography (ERCP) under total intravenous anesthesia.Methods Ninety patients,47 males and 43 females,aged 18 years old or more,falling into ASA physical status Ⅰ-Ⅲ,undergoing ERCP under total intravenous anesthesia from July 18,2016 to August 31,2016 in Ruijin Hospital were included in our study.General information and intraoperative situation of patients were collected.The incidence of PONV within 24 h was recorded.A multivariate Logistic regression model was performed to analyze the independent risk factors of PONV after ERCP under total intravenous anesthesia.Results In total,90 patients were included in this study,with 33 patients (36.7%) suffering PONV.Logistic analyses showed that female (OR=3.73,95%CI 1.36-10.27),history of PONV/motion sickness(OR=4.39,95%CI 1.40-13.76),and serum-amylase greater than 3 times the normal value (OR=5.22,95%CI 1.30-20.95) were the independent risk factors for PONV after ERCP under total intravenous anesthesia.Conclusion Female,history of PONV/motion sickness,and serum-amylase greater than 3 times the normal value were the independent risk factors for PONV after ERCP under total intravenous anesthesia.

Objective To research and compare the effectiveness distinction between applying exclusive salcatonin and combining salcatonin and Xianling Gubao to treat osteoporotic pain patients.Methods 84 cases of osteoporotic pain patients in our hospital from Feb.2013 to Feb.2014 were selected and divided into two groups.Patients in treatment group were received combination of salcatonin and Xianling Gubao treatment.Patients in control group received salcatonin treatment only.To apply VAS judge the pain degree of patients.The clinical performance and results were observed and recorded.The degree of satisfaction of patients was surveyed through questionnaire.Results The VAS of patients in treatment group was (2.17 ±1.02), while it was (4.26 ±1.37)in control group.There were 15 cases of excellent,and 23 cases of efficacious in treatment group and the total effective rate was 90.48%.There were 10 cases of excellent,and 19 cases of efficacious in control group and the total effective rate was 60.95%.The degree of satisfaction of patients in treatment group was 95.24%,while it was 71.43% in control group.Conclusion Combining salcatonin and Xianling Gubao has better clinical effect than applying exclusive salcatonin in treatment of patients with osteoporotic pain.The combination of them could quicken the speed of recovery and increase the degree of satisfaction of patients.

[Objective]To initially approach the role of lumbar facet joint derived inflammatory factors in degenerative lumbar spinal canal stenosis. [Methods]Totally 75 cases of degenerative lumbar spinal canal stenosis(LSCS)(n=41)and lumbar intervertebral disc herniation(LDH)(n=34) undergoing posterior lumbar spinal surgery in our department were evaluated in terms of the extent of degenerative arthrosis according to the Weishaup grading criteria.The grading of backleg pain,melosalgia and functional impairment were recorded.The excised lumbar facet joints were collected as species.The content of interleukin-1? and tumor necrosis factor-? in the species were determined by ELISA.[Results]There was no TNF-? detected in both of the two groups.More IL-1? was detected in degenerative lumbar spinal canal stenosis group than that in lumbar intervertebral disc herniation group.It was demonstrated that the content of IL-1? in the species increased as the degeneration of lumbar facet joint sharpened.IL-1?-positive cases in degenerative lumbar spinal canal stenosis group showed higher grading of backleg pain,melosalgia and functional impairment.[Conclusion]The cartilage of degenerative lumbar spinal canal produced more IL-1?.Lumbar facet joint derived inflammatory factors might be one of the reasons that cause backleg pain and melosalgia and functional impairment in degenerative lumbar spinal canal stenosis patients.

To investigate the monitoring of acute rejection of porcine pancreaticoduodenal allografts, 12 pigs were subjected to pancreaticodudenal allografts, among them 6 pigs received triple therapy for 7 days. Serial biopsies were performed in pancreas allografts for diagnosis of pancreatic acute rejection. Amylase secretion was determined periodically.The levels of TNF ? and NO in serum were serially monitored.The results showed that amylase secretion decreased,TNF ? and NO concentration in serum increased significantly as acute rejection was detected histo pathologically in pancreatic grafts. It suggested that monitoring of amylase secretion, serum TNF ? and NO concentration could be used to decide the proper time of performing biopsy for early diagnosis of acute rejection of the pancreas allograft.

Objective:To investigate the feasibility of marrow stromal cells/ceramic bovine bone/hydrogel in the repair of autogenous calvarial defects. Methods:The complex of ceramic bovine bone,hydrogel,rhBMP-2,TGF-? and marrow stromal cells(MSCs) induced by minerlization inducing medium was prepared and applied in the repair of autogenous calvarial defects in 8 SD rats(group A).Ceramic bovine bone /MSCs was used as control in another 8 SD rats(group B). 4 and 8 weeks after operation 4 rats in each group were sacrificed, the osteogenesis was examined by HE staining,modified Mallory's trichrome staining and image analysis.Results:4 weeks after operation, osseous plerosis can be seen in both groups, the quantity of bone-like tissues formation(?m~2) in group A and group B was 6 813.09?96.32 and 3 839.25?104.52(P

With the continuous development of imaging techniques,patients with congenital heart dis-ease,especially in complicated congenital cardiac malformations,have more choices for detection methods of ear-ly diagnosis. Nowadays,the widely used diagnostic imaging methods included echocardiography,CT,MRI,cardi-ac catheterization,and prenatal ultrasound examination. Clinicians should choose the appropriate means of ima-ging test depending on the situation. This article will focus on the clinical application of imaging diagnosis in complicateol congenital heart disease.

Objective: To study the clinical features of depressive patients consulting neurological clinics in a general hospital and the therapeutic effect of Paroxetine in treatment of depression in this group Methods: 46 patients with depressive symptoms were collected consecutively, 24 among them were treated by Paroxetine The changes of scores of HAMD and Clinical Global Impression(CGI) before and after treatment were used to assess the therapeutic effect Results: Patients complained a variety of somatic symptoms involved nearly all systems, After four weeks’treatment of Paroxetine, both HAMD and CGI scores decreased dramatically The change of HAMD became significant at the end of the first week of treatment Conclusion: Depressive patients consulting general hospital have many somatic complaints Paroxetine has both anxioletic and antidepressant effects in this group of patients

Objective To summarize the clinical experience of retroperitoneoscopic surgery in the treatment of adrenal tumors. Methods Retroperitoneoscopic resection of adrenal tumors was performed in 35 patients(12 males and 23 females) aged 25~72 years(mean,49.8 years).The tumor was left-sided in 15 patient and right-sided in 20 patients.There were 5 cases of pheochromocytoma,10 cases of primary hyperaldosteronism,2 cases of Cushing's disease,2 cases of paraganglioma,and 16 cases of dysfunctional adenoma.The operation was performed through 3 lumbar ports.A self-made water-filled balloon dilator was employed to expand the retroperitoneal cavity,and then the tumor was removed by using a harmonic scalpel.Results The procedure was successfully completed in all the 35 patients,with no need of conversions to open surgery.The intraoperative blood loss was 10~500 ml(mean,74 ml).No blood transfusion was required.The operation time was 45~200 min(mean,90 min).Conclusions Retroperitoneoscopic adrenalectomy has advantages of satisfactory effects,short operation time,little blood loss and invasion,and quick recovery.It can replace most of traditional open operations for adrenal tumors.

To investigate the therapeutic effect of ascitic fluid ultrafiltration refusion and perfusion of ?-elemene milk into abdominal cavity on carcinomatous ascites. Seventy-two cases of malignant tumor complicated with carcinomatous ascites were randomized to two groups: group A treated with perfusion of ?-elemene milk into abdominal cavity after ascitic fluid ultrafiltration refusion and group B with perfusion of ?-elemene milk into abdominal cavity after letting-out of ascitic fluid by regular and repeated abdominal paracentesis. Therapeutic effect was compared and ascitic protein (AP) , plasma albumin(PA) content, urine volume and Karnofsky scores were observed before and after treatment as well as the toxic and side effects.In group A, 9 cases were completely relieved (CR) , 22 partially relieved (PR) , 12 not changed (NC) and the effective rate was 86.1%;in group B,4 were CR, 20 PR, 12 NC and the effective rate was 66.7% respectively, the differences being significant (P