Objective To find an internal quality controlling method for 17-OHP determination by time-resolved fluoroimmuno-assay .Methods 20 quality control data were collected .The data were analyzed by using L-J method ,instant method and improved instant method .Results were used for the construction of quality control charts .Result The first three quality control data had a great impact on the following judgments of internal quality controlling when instant method was used .The subsequent results might be false acceptance .Improved instant method could effectively reduce the situations of false run-away and false acceptance ,which was suitable for the internal quality control of 17-OHP determination by time-resolved fluoroimmunoassay in newborn screening . Conclusion There are many steps of manual operations in 17-OHP determination of time-resolved fluoroimmunoassay .The details of these operations have great impacts on the experimental results .Thus ,the operations of 17-OHP test should be specified and exe-cuted strictly according to requirement .

Objective To discuss the causes of misdiagnosis and therapeutic errors of tracheobronchial foreign bodies.Methods 139 cases of tracheobronchial foreign bodies hospitalized in our department during 5 years were reviewed. Retrospective analysis was performed, on statistics of case history, clinical manifestation, location of foreign bodies, the time of initial diagnosis and the time for definite diagnosis.Results 89 cases (64 03%) were misdiagnosed, and were given inappropriate treatments. The main causes include ignored or vague history of foreign body inhalation, the atypical or complex clinical manifestations, and underestimation of the importance of bronchoscopy.Conclusion We should pay more attention to history of foreign body ,bronchoscopy examination action in diagnosis and treatment and more aware of the various manifestations of tracheobronchial foreign bodies,these could contribute to decrease the misdiagnosis and therapeutic errors.

Objective To recombine the tissue plasminogen activator (TPA) gene and establish the expressing model in vitro to provide both theoretical basic and technical guidance for gene therapy of ischemic heart disease and prevention of postoperative vessel re-stenosis. Methods Expression vector pcDNA3.1 TPA was constructed, and transfected into Chinese hamster ovary(CHO) cells. Then the exogenous TPA expression was observed. Results The expression quantity of TPA in transfected CHO cells was higher than that in non-transfected cells. The exogenous TPA activity was 12 296 IU/10 6cell/24hr in the transfected cells, when measured by chromogenic substrate assay, while that of non-transfected cells was 3 176IU/10 6cell/24hr. The TPA quantity of non-transfected cells was 9 608 ng/10 6cell/24hr, when measured by enzyme-linked immunosorbent assay (ELISA), while that of transfected cells was 586 172 ng/10 6 cell/24hr, and the latter was the 60 times of the former. Conclusions When pcDNA3 1(+)TPA was transfected into CHO cells, exogenous TPA can efficiently be expressed, which provides theoretical basic for TPA clinical gene therapy.

The clinical data of 97 patients who underwent laparoscopic hepatectomy from July 2007 to November 2014 in the First Affiliated Hospital of Wenzhou Medical University was collected.Of the 97 patients,46 cases were diagnosed with hepatolithiasis,20 with primary liver cancer,26 with benign liver tumor,1 with liver abscess,and 4 with metastatic liver cancer.The surgical approaches included laparoscopic left hepatectomy (n =16),left lateral lobectomy (n =35),right hepatectomy (n =2),right posterior lobectomy (n =1),hepatic wedge resection (n =42),and left lateral lobectomy with segment Ⅶ resection (n =1),and 4 cases underwent multi-visceral resection.88 patients underwent laparoscopic hepatectomy and 9 patients were transferred to open hepatectomy.Postoperative complications included bile leakage (n =9),abdominal cavity infection (n =7),ascites (n =1),wound infection (n =1),pleural effusion (n =3) and pulmonary infection (n =1).There was no perioperative death.Taken together,laparoscopic hepatectomy is feasible and efficient for treating liver diseases and it has the advantages of minimal invasion and rapid recovery.

Objective To investigate the effects of captopril on reperfusion arrhythmias in atherosclerotic rabbits.Methods A total of 32 New Zealand white rabbits were randomly divided into the control group,cholesterol food group(CF group) and captopril plus cholesterol food group(Cap group).The levels of endothelin(ET),malonaldehyde(MDA) and the content of nitric oxide(NO) were measured in different stage.The duration of ventricular tachycardia and the incidence of ventricular premature beats in 30 minutes after reperfusion were detected.Results The levels of ET and MDA were significantly decreased and the content of NO were significantly increased in the Cap group compared with that of the CF group.The average duration of ventricular tachycardia and the incidence of ventricular premature beats in 30 minutes after reperfusion were significantly reduced in the Cap group compared with the CF group.Conclusion The incidence of arrhythmias induced by the ischemia reperfusion injury may be decreased with the treatment of captopril in the atherosclerotic rabbits.

Objective To study the diagnosis and surgical treatment of Mirizzi syndrome (MS).Method The clinical data of patients with Mirizzi syndrome treated in our center from July 2001 to July 2011 were retrospectively studied and the diagnostic methods,operative strategies and outcomes of surgical treatment were analyzed.Results Mirizzi syndrome (MS) was identified in 56 out of 13800patients who received cholecystectomy (0.4％). MS was diagnosed preoperatively in 30 patients (53.6％).There were 29 patients with Mirizzi syndrome type Ⅰ,17 patients with type Ⅱ,9 patients with type Ⅲ,and 1 patient with type Ⅳ using the Csendes's classification.In two patients (3.6％) coincidental gallbladder carcinoma was detected.An initial laparoscopic approach was attempted in 33patients,and 16 were converted to open surgery.The remaining 23 patients underwent open operation.Surgical procedures included cholecystectomy,choledochotomy and T-tube insertion,simple closure and drainage (via T tube) of the biliary fistula,Roux-en-Y hepaticojejunostomy,radical resection of gallbladder and hepaticojejunostomy.Inadvertent bile duct injury occurred in 2 patients who had an initial laparoscopic approach for a preoperative undiagnosed MS. Postoperative morbidities included biliary leak (n =4) and residual common bile duct stone (n=2).All patients recovered completely and there was no hospital mortality.Conclusions Preoperative diagnosis of Mirizzi syndrome is still challenging despite the availability of multiple imaging modalities.Open surgery remains the standard of care,although laparoscopic treatment may be used in selected patients,especially for type Ⅰ Mirizzi syndrome.Patients with Mirizzi syndrome should be managed differently,basing on intraoperative findings and the type of Mirizzi syndrome.

Objective:To analyze gene expression profiling of left ventricular myocardium in patients with chronic congestive heart failure(CHF)caused by rheumatic heart disease(RHD)with the normal controls in order to identify CHF associated target genes. Methods:The gene expression profiles of left ventricular myocardium from patients with CHF by RHD and normal controls were obtained from six human whole-genomic oligonucleotide microarrays(Affymetrix HG U133 Plus 2.0 GeneChip). GeneSpring software was used to identify the differentially expressed genes in both groups,and bioinformatic analysis was applied to analyze the target genes associated with CHF. Real-time PCR was carried out to validate the expression of three target genes. Results:We identified 102 target genes associated with CHF which were classified into 7 gene clusters. Microarray results were further confirmed by real time PCR for three genes. ATF3 was markedly down-regulated,IGFBP2 and NPPB were notably up-regulated in the left ventricular myocardium samples from CHF patients. Conclusion:A lot of differentially expressed genes,obtained by using the whole-genomic expression profiling technology,might be a contributory factor for the initiation and progression of CHF and it helpful for the understanding of underlying pathophysiological implications. Further investigation on these genes would provide a strategy to identify genetic markers and molecular events associated with CHF caused by RHD.

Objective To investigate the role of RhoA /Rho-kinase pathway in rat models of left heart disease-as-sociated pulmonary hypertension ( PH-LHD) .Methods Twenty male SD rats (3-4 week-old, 90-100 g) were randomly divided into two groups (10 rats in each group):the group C ( control group) with sham operation, and group H ( pulmo-nary arterial hypertension) .The rat model of left heart disease-associated pulmonary hypertension was established by supra-coronary aortic banding in the group H, and the sham surgery was applied for the rats in the group C ( The titanium clip was fixed at the mediastinal tissue adjacent to the artery rather than the ascending aorta).On day 60 after the operation, the cardiac functions, including right ventricular systolic pressure and pulmonary artery pressure were evaluated.After that, all rats were sacrificed and treated with cardiopulmonary lavage in vivo until the lung became white.Then the left lung tissues were fixed in 4%paraformaldehyde for pathological observation while the right lung tissues were frozen for mRNA detec-tion.Results Compared with the group C, both ventricular systolic pressure and pulmonary artery pressure in the group H were increased significantly (P<0.01).Pathological data demonstrated that the pulmonary artery walls in H group were much thicker than that in the group C.Moreover, vascular wall hypertrophy index in the group H was increased greatly compared with that in the group C (P<0.01).QPCR data showed that mRNA levels of Rho kinase, RhoA and ET-A R in the group H were up-regulated compared with the group C ( P<0.01) .Conclusions Rat model of left heart disease-asso-ciated pulmonary arterial hypertension can be successfully established by supracoronary aortic banding.Rho-kinase-media-ted pathway may contribute to the pathogenesis and progress of left heart disease-associated pulmonary arterial hypertension.

AIM: To investigate the effect of microwave ablation of liver cancer on the cellular immunity in mice. METHODS: A C57BL/6J mouse model of liver cancer was established by subcutaneous injection of Hepa 1 - 6 cells. The tumors were subjected to microwave ablation under the ablation condition of 45 ℃, 50 ℃, 55 ℃ or 60 ℃ for 180 s. The CD4~+ T cells, CD8~+ T cells and natural killer cells (NK) in peripheral blood were detected by FACS. The cytotoxicity of splenic NK and splenic cytotoxic T lymphocytes (CTL) activated by inactivated Hepa 1-6 cells was assayed by LDH method. RESULTS: The proportions of CD4~+ T cells, CD8~+ T cells and NK cells in peripheral blood in 50 ℃ and 55 ℃ group at 21 d after ablation were significantly increased and that of NK cells in 60 ℃ group was significantly increased. There was no significant difference between those in group 42 d after ablation and control. The cytotoxicities of splenic CTL and NK cells in 50 ℃ and 55 ℃ groups at 21 d or 42 d after ablation were significantly increased, and they were much higher than those in 45 ℃ group at the same time. The cytotoxicities of splenic CTL in 50 ℃ and 55 ℃ groups at 21 d after ablation were much higher than that in 60 ℃ group at the same time. CONCLUSION: Under a certain ablation temperature, microwave ablation of liver cancer promotes the cellular immunity.

ObjectiveTo introduce a new technique to create a pulmonary valve biorifice for reconstruction of right ventricular outflow tract in tetralogy of Fallot (TOF),and to summarize its initial clinical experience and therapeutic effect.MethodsThe new technique regarding reconstruction of right ventricular outflow tract with a pulmonary valve biorifice was used in a total of 53 TOF cases (the observation group).The conventional technique regarding reconstruction of right ventricular outflow tract was used in other 50 TOF cases (the control group).The clinical dates of all cases were reviewed retrospectively.ResultsThe ages,weights,cardiopulmonary bypass time,cardiac arrest time,as well as the post operation ventilation support time were not different significantly between two groups.Compared with the contrul group,patients from the observation group had shorter duration of ICU stay.After operation,in the observation group,only 2 cases had large amount of pleural effusion,1 case meddle,and 8 cases little amount of pleural effusion; whereas,in the control group,the corresponding numbers were 1,5 and 17,respectively.At the time point of 1 week after operation,all patients were rechecked by echocardiography,no pulmonary valve stenosis was found.Moderate pulmonary valve regurgitation was found in 8 cases,mild regurgitation in 15 cases from the observation group; and severe regurgitation in 3 cases,moderate regurgitation in 17 cases,and mild regurgitation in 16 cases from the control group.A total of 33 cases from the observation group were rechecked at the time point of half year after operation,and moderate - mild pulmonary regurgitation were found in 3 cases.A total of 18 cases of them were rechecked 1 - year latter,no pulmonary regurgitation was found.ConclusionsThe new technique to create pulmonary valve biorifice can reduce the pulmonary valve regurgitation and postoperative pleural effusion,and improve the early outcomc.