2.Recombination expression and immunoprotective effects of the P6 outer membrane protein of nontypeable Haemophilus influenzae
Xiangpeng CHEN ; Jian WANG ; Lili LI ; Jing CHEN ; Zhenlong ZHANG
Chinese Journal of Microbiology and Immunology 2010;30(4):349-354
Objective To clone and express recombinant outer membrane protein P6, determine its optimal expression conditions and to investigate the immunoprotective effects of the P6 protein on mice. Methods The P6 gene of nontypeable Haemophilus influertzae(NTHi) was amplified by PCR from the NTHi genome and cloned into expression vector pET-32a (+) to generate the pET-32a-P6 recombinants. They were confirmed by nuclease digestion and sequence analysis. The verified recombinant was transformed into E. coli BL21 (DE3). Its optimal expression conditions were determined such as engineering strains, the concentration of IPTG, inducing temperature, inducing time, different medium etc. The recombinant protein was purified by Q Sepharose~(TM) XL ion exchange and gel filtration chromatography. The protein was analyzed by SDS-PAGE, Western blot and sequencing. BALB/c mice were immunized with recombinant protein be-fore challenged by NTHi through intraperitoneal injection. Then the mortality rate of different group was com-pared. Results The recombinant P6 of NTHi was successfully constructed and expressed in E. coli at a rel-atively high level. The purity was up to 95% after purification. The relative molecular mass of the protein is 14 145. 848. The recombinant protein was confirmed to show specific reaction on the antiserum through Western blot. The animal experiments showed the mortality rates of immunization groups were significantly lower than that of the control group (P < 0.05). Conclusion The successful expression of the recombinant P6 will be very helpful for the further study on development of vaccine, its purified, immunological activity and antibody preparation.
3.Non-enhanced CT predicting nonbronchial systemic arterial supply in patients with hemoptysis
Zhensheng LIU ; Jiaxiang WANG ; Xiongwei KUANG ; Zhenlong XUE ; Cheng LI
Chinese Journal of Interventional Imaging and Therapy 2009;6(4):363-366
Objective To assess the prediction value of nonbronchial systemic arterial supply in hemoptysis patients with non-enhanced CT. Methods Fifty-six consecutive patients with hemoptysis underwent non-enhanced CT. Thickness of pleural adjacent to parenchymal lesion larger than 3 mm was regarded as index of nonbronchial systemic arterial supply. Conventional angiography was used as the standard of reference. CT findings were compared with those of conventional angiography. The sensitivity, specificity, and accuracy of CT for predicting nonbronchial systemic arterial supply were assessed. Results The sensitivity, specificity and accuracy of CT for predicting nonbronchial systemic arterial supply were 72.73%, 95.00% and 91.11%, respectively. Sensitivity was higher when nonbronchial systemic arterial supply located in superolateral and posterolateral lung, and lower in anteromedial and inferior lung. Specificity and accuracy were high for predicting nonbronchial systemic arterial supply in every locations. Conclusion Non-enhanced CT can predict nonbronchial systemic arterial supply in patients with hemoptysis, which is helpful for selecting angiography and embolization.
4.Significance of the MELD scoring system in surgical treatment of obstructive jaundice
Zhenlong WANG ; Yong YU ; Boyi CHEN ; Chencai LI ; Rong LI
Chinese Journal of Hepatobiliary Surgery 2014;20(7):503-506
Objective To prospectively study the clinical significance of the MELD scoring system in surgical treatment of obstructive jaundice.Methods 112 patients with obstructive jaundice who were admitted into our hospital from January 2009 to December 2013 were divided into two groups:group A (Stage Ⅰ PTCD and stage Ⅱ open operation,n =53) and group B (1 stage open surgery,n =59).The amount of intraoperative bleeding blood loss,operation time,postoperative complications,duration of hospitalization,mortality rate,and the changes in liver function after surgery were compared between the two groups.Results The differences in the liver function index of the two groups on the same postoperative date were significantly different (P < 0.05).The liver function of group A recovered faster than group B.Patients in group A with a MELD < 10 points stayed in hospital significantly longer when compared with patients in group B.For patients in group A with MELD > 10 points,the operation time,bleeding volume,postoperative complications and hospitalization were significantly less than the patients in group B (P < 0.05).There were 3 patients (group B) who died with MELD ≥ 20 points after operation.Conclusions In patients with obstructive jaundice with a MELD score greater than 10 points,especially those with a score equal to or greater than 20,PTCD should be performed first to relieve biliary tract obstruction,followed by a stage Ⅱ open surgical operation after the liver function had improved.MELD had important clinical significance in the evaluation of operation risk in patients with obstructive jaundice.
6.Analysis of CYP2C19 metabolism phenotypes distribution of 76 patients with clopidogrel treatment in Huangshi
Zhongliang ZHU ; Congrong LI ; Hongliang WANG ; Zhenlong LI
International Journal of Laboratory Medicine 2014;(11):1410-1411,1414
Objective To investigate the distribution of CYP2C19 metabolism phenotype in patients with clopidogrel treatment in Huangshi.Methods The peripheral blood samples were obtained from 76 patients with clopidogrel treatment in Cardiovascular Department.CYP2C19 genotypes were determined by the gene chip,CYP2C19 metabolism phenotype were investigated and com-pared with the data of healthy Han Chinese from the published papers.Results There were three kinds of metabolizers about the metabolism phenotypes of CYP2C19,which were extensive metabolizers,intermediate metabolizers and poor metabolizers.In the 76 patients,the ratios of these metabolism phenotypes were 39.47%,44.74% and 15.79%,respectively.The result coincided with the healthy Han Chinese.Conclusion The clinical individualized medication of healthy Han Chinese could be refered to the clinical indi-vidualized medication of patients with clopidogrel treatment in Huangshi.
7.Sentinel lymph node biopsy in patients with operable breast cancer after neoadjuvant systemic therapy
Kai LUO ; Zeping YU ; Zhenlong WANG ; Bo WU ; Jieshou LI
Journal of Medical Postgraduates 2003;0(08):-
Objective: To investigate the feasibility and accuracy of sentinel lymph node biopsy(SLN) following neoadjuvant chemotherapy.Methods:Sentinel lymph node biopsy was evaluated following two or three cycles of neoadjuvant chemotherapy with pharmorubicin /cyclophosphamide/5-FU(CEF,48 patients) or pharmorubicin /cyclophosphamide / taxotere(CET,12 patients).Two ml 1% methylene blue was subcutaneously injected at 4 sites of the skin over the tumor before radical operation.Pathohistological results were assessed for all the specimen including blue lymph node and other axillary lymph node(ALND). Results:Among the 66 patients SLN was identified in 60 of them(90%).Residual metastatic disease was identified in 23 cases on SLN,including 15 positive and 8 negative node on ALND,37 cases were completely negative on SLN including 35 negative and 2 positive node on ALND,and SLN was falsely negative in 2 cases(8%).Conclusion:The success rate of sentinel node identification and the predict ability of the SLN for the pathologic status of the adjacent non-SLNs do not seem to be altered after neoadjuvant therapy.
8.The surgical strategy for tetralogy of Fallot with pulmonary atresia
Minhua FANG ; Huishan WANG ; Hongyu ZHU ; Zengwei WANG ; Zhenlong WANG ; Chunzhen ZHANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(9):539-541
Objective To evaluate the indication and surgical technique for treating tetralogy of F allot with pulmonary atresia (TOF-PA).Methods From June 1984 to June 2009,66 patients with TOF-PA underwent 69 operations.Among them,34 were males and 32 females.Their age ranged from 6 months to 29 years.The anatomic characteristics of TOF-PA included 31 cases of Type Ⅰ,14 Type Ⅱ,12 Type Ⅲ and 9 Type Ⅳ.The operations included palliative aorto-pulmonary shunts in 11 cases,one-stage unifocalization with unpatched VSD in 2 cases,one stage complete repair in 40 cases,one-stage unifocalization with VSD repair in 13 cases,and delayed intracardiac repair after shunt procedures in 3 cases.Results There were 6 early deaths,including 1 death happened after aorta-pulmonary shunt and 5 after complete repair.The causes of death were severe low cardiac output in 3 cases,respiratory failure in 1,multiorgan function failure in 1 and severe wound infection with endocarditis in 1 after aorta-pulmonary shunt.The postoperative oxygen saturation of the patients undergone shunt and one stage unifocalization with unpatched VSD increased to 82% ~ 91%.The postoperative ratio of right ventricular pressure/left ventricular pressure after complete repair was < 0.5 in 31 cases,18 cases were between 0.5 and 7 cases > 0.75.47 patients were followed up from 3 months to 15.5 years.The heart function(NYHA) of 44 patients were in class Ⅰ or Ⅱ and 3 in class Ⅲ or Ⅳafter operation.Conclusion The surgical strategy for TOF-PA mainly depends on the anatomic characteristics of the pulmonary and aortopulmonary collateral arteries.An individualized approach based on the anatomy of the pulmonary circuits permits a better result in the patients with TOF-PA.Patients with well developed pulmonary arteries should undergo one stage complete repair as early as possible.
9.The results of the Fontan procedure in patients with visceroatrial heterotaxy syndrome and complex cardiac anomalies
Minhua FANG ; Huishan WANG ; Hongyu ZHU ; Zengwei WANG ; Zongtao YIN ; Zhenlong WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;28(9):519-521
Objective To assess the results of the Fontan procedure in patients with visceroatrial heterotaxy syndrome and complex cardiac anomalies.Methods From April 2002 through December 2010,25 patients (male 10,female 15) had undergone the Fontan procedure for heterotaxy syndrome or atrial isomerism and complex congenital heart disease 28 times.Median age at operation was (9.1 ± 5.5) years (2 to 18 years).Heterotaxy syndrome were associated with right atrial isomerism (n =18) or left atrial isomerism (n =7),asplenia (n =13) or polysplenia (n =7),double inlet of left ventricle (n =15),double inlet and outlet of left ventricle (n =5),double outlet right ventricle with pulmonary atresia (n =2) and with pulmonary stenosis (n =1),tricuspid or mitral atresia (n =2).A bidirectional cavopulmonary shunt was performed in 8 patients (bilateral in 3 patients).A cavopulmonary shunt placement,so-called Kawashima operation,was performed in 4 patients.An extracardiac conduit Fontan connection was pefformed in 15 patients and intracardiac lateral tunnel Fontan connection in one patient.Results 2 patients died in hospital caused by ventricular failure.Five patients developed early postoperative atrial arrhythmias and 2 patients had sinus node dysfunction.Mean arterial oxygen saturation at discharge was 0.86 ± 0.07 (range,0.78 to 1.00).Follow-up (range,0.5 to 7 years) was available on 15 patients.Mean arterial oxygen saturation was 0.82 ±0.08 (range,0.68 to 0.97).Ventricular function was normal in 13 patients (EF range,0.50 to 0.66) and depressed in 2 patients.Four patients had a junctional rhythm.Conclusion The Fontan procedure was still the main procedure for patients with visceroatrial heterotaxy syndrome and complex cardiac anomalies,which can reach satisfactory early and medium-term results.The choice of Fontan procedure,extracardiac conduit Fontan connection,aggressive treatment of concomitant malformations were essential to improve the outcomes.
10.The analysis of results in the young children with tetralogy of Fallot: one-stage versus staged repair
Minhua FANG ; Huishan WANG ; Zengwei WANG ; Zhenlong WANG ; Chunzhen ZHANG ; Bo LI
Chinese Journal of Thoracic and Cardiovascular Surgery 2017;33(5):267-270
Objective The study aimed to evaluate the short and middle term results in the patients with tetralogy of Fallot(TOF) after one-stage repair and staged repair.Methods A total of 459 TOF younger children less than five-year-old between January 2009 and December 2013 had received surgical repair,including 416 patients by one-stage repair(group Ⅰ)and 43 patients by staged repair(group Ⅱ).Among them,245 were male and 214 were female.The average repair surgical age was 27.8 ranged from 4 to 60 months,average palliation age was 15.4 ranged from 3 to 40 months.23 perioperative and follow-up parameters were assessed including sex,age,weight,preoperative clinic symptom,ratio of McGoon,pulmonary artery index,Z score of pulmonary annulus,cardiopulmonary bypass time,aortic cross-clamping time,type of VSD repair,type of RVOT procedure,Peak RV/LV pressure radio,RV-PA pressure grade,mortality,severely low cardiac output syndrome,hypoxemia,extubation time,ICU time,fellow-up time,left ventricular eject faction,RV index of myocardial performance (MPI),tricuspid annular plane systolic excursion(TAPSE) and pulmonary regurgitation.Results Compared with group Ⅰ,age and weight were significantly lower in patients in group Ⅱ[(19.1 ± 16.4) months vs.(21.1 ± 11.2) months,P < 0.05) and(19.1 ±16.4) kgvs.(21.1 ±11.2) kg,P <0.01].The Z score of pulmonary annulus of patients in group Ⅱ were significantly less than those in group Ⅰ (-3.69 ± 2.36 vs.-2.50 ± 1.95,P < 0.01).The ratio of repairing VSD by RV incision and using TAP in patient of group Ⅱ were significantly more than those in group(27/43 vs.71/413,P <0.01),(41/43 vs.221/413,P <0.01).There was no difference of mortality,complication,extubation time and ICU time bewteen two groups.All patients followed up 12-52 months,there was no difference of LVEF,MPI and TAPSE between two groups.However,the severity of pulmonary regurgitation in patients of group Ⅱ was significant more than those of group Ⅰ (47.6% vs.32.1%,P < 0.01).Conclusion The early and mid-term results in the the young children patients with TOF after one-stage repair or staged repair were good.Although the prior palliative shunt could promote the development of the hypoplasia pulmonary arteries in the young children patient,it may related to the technique of repairing operations and the postoperative pulmonary regurgitation.