Effects of poly(A) tails with different lengths on mRNA expression in vitro and stability of transcription template in Escherichia coli / 中华微生物学和免疫学杂志

Objective:To investigate the effects of poly(A) tails with different lengths on mRNA expression in vitro and the passage stability of transcription template with poly (A) tail in Escherichia coli ( E. coli). Methods:Plasmids with poly(A) tails of 38, 60, 103, 125 and 126 (60 nt+ 6 nt spacer+ 60 nt) nt were designed and constructed. Then the plasmids were linearized by single enzyme digestion and used as transcription template for preparing enhanced green fluorescent protein (EGFP)-mRNA. EGFP-mRNA containing poly(A) tails of different lengths were transfected into 293T cells and the expression of EGFP was detected by flow cytometry. As to stability test, the template plasmids with poly (A) tail of 125 and 126 nt were transformed into E. coli TransStbl3 and Top10 competent cells. Seven clones were selected for culture and plasmid extraction, and then the plasmids were digested by restriction enzyme and detected by capillary electrophoresis. For passage stability, three correctly sequenced clones of each group were selected for continuous passage at 37℃, and the plasmids were extracted and digested every two generations for capillary electrophoresis. At the same time, the correctly sequenced clones of 125 nt group were also passaged at 30℃, and the plasmids were also extracted and digested every two generations for capillary electrophoresis. Results:The transcription templates with poly(A) tail of different lengths were successfully constructed. Flow cytometry showed that the fluorescence expression of the template plasmids with poly (A) tail of 103 and 125 nt were significantly higher than that of 38 and 60 nt. The fluorescence expression of the plasmid with poly (A) tail of 126 nt was significantly higher than that of all other groups. The percentages of stable sequences of the template plasmid with poly(A) tail of 125 nt in TransStbl3 and Top10 competent cells were 76% and 91%, respectively. The results of continuous passage showed that poly(A) tail of 125 nt could be stable to the 4th generation at 37℃ in both TransStbl3 and Top10 competent cells, and stable to the 16th and 10th generations at 30℃. The percentages of stable sequences of the template plasmid with poly(A) tail of 126 nt in TransStbl3 and Top10 competent cells were 95% and 48%, respectively. The results of continuous passage showed that poly(A) tail of 126 nt could be stable to the 12th generation at 37℃ in both TransStbl3 and Top10 competent cells.Conclusions:The length and composition of poly(A) tail in mRNA affected the expression of target protein. Adding a spacer with a length of 6 nt to poly(A) tail and low temperature culture were both helpful to improve the stability of the template plasmid, which provided a reference for the design and preparation of in vitro transcription template of mRNA vaccine.

Improvement to colon interposition after esophagectomy / 中华胸心血管外科杂志

Objective To explore the improving safety measures of colon interposition after esophagectomy .Methods From January 2003 to December 2014, 65 cases of colon interposition after esophagectomy were performed, and some improve-ments were made in the methods and procedures .Mainly including: Preoperative evaluation to first operation; Selection of co-lon segment; Simplify vascular anatomy;Cervical anastomosis was replaced by intrathoracic anastomosis;Modified anastomosis sequence; Strengthen preoperative preparation.Results There were no perioperative deaths.Only 2 patients with cervical fis-tula, due to malnutrition automatically discharged.The rate of anastomotic leakage on neck were 27.77%(10/36) and 6.89%(2/29) in the chest.2 cases were completed intestinal obstruction after jejunostomy , 1 cases of volvulus and 1 cases of intus-susception confirmed by reoperation.Conclusion Colon interposition after esophagectomy is a very important way of surgery. The operation process is complex, but as long as mastering the point of surgery, improving the surgical procedures, the good effect can be obtained and greatly improve the safety of the operation .

Strategies of surgery treatment for cervical esophageal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the methods and means of surgical treatment for cervical esophageal cancer in order to get the best efficacy.</p><p><b>METHODS</b>Retrospective investigation and analysis were carried out in 137 patients with cervical esophageal cancer undergoing operation in our hospital from January 2005 to December 2013. According to lesion locations, these cases were divided into two groups: larynx preservation group(Group A) and laryngectomy group(Group B). Surgery options of Group A included inversion esophagectomy without thoracotomy, three-incision esophagectomy, end to end anastomosis of cervical esophagus after local resection. Group B included gastric-pharyngeal anastomosis, pharyn-esophagus plasty, colon replacement of the esophagus or jejunal interposition. All the patients received postoperative adjuvant therapy.</p><p><b>RESULTS</b>There were no perioperative deaths. The main postoperative complications included anastomotic fistula in 20 cases(14.6%), postoperative massive bleeding after inversion esophagectomy in 3 cases, chylothorax in 1 case, anastomotic stenosis in 9 cases, severe gastroesophageal reflux in 6 cases and serious aspiration pneumonia in 2 cases. All these patients had complete resolution of disease with good postoperative quality of life. The 1-year, 3-year and 5-year survival were 73.7%, 48.4% and 26.8% respectively. The most important causes of postoperative death included local recurrence, cervical or mediastinal lymph node metastases, cachexia and multiple organic metastasis.</p><p><b>CONCLUSION</b>It is critical to select reasonable methods of operations to reduce postoperative complications and to receive proper adjuvant therapy.</p>

An analysis of 5-year survival after pneumonectomy and adjuvant chemotherapy in lung cancer / 中国癌症杂志

10.3969/j.issn.1007-3969.2013.05.010

Treatment for prosthetic valve endocarditis after mitral valve replacement therapy / 中国综合临床

Objective To review the experience of treatment for prosthetic valve endocarditis(PVE)after mitral valve replacement(MVR)in 16 cases.Methods From September 1979 to December 2010,16 patients were diagnosed as PVE after MVR by modified Duke University diagnostic criteria.There were 10 males and 6 females.Their ages ranged from 19 to 55 years old(mean 28 years old).The incidence of PVE was 0.97％(16/1657)after MVR.Blood culture positive was in 5 cases.Medical treatment alone was performed in 10 patients who were treated by using Penicillin or Vancomycin with other broad-spectrum antibiotics,using Fluconazole and Amphotericin if necessary.Combined medical and surgical management were performed in 6 cases.One emergency case was operated because of obstinate heart failure.Five cases underwent operation after adequate antibiotics treatment and general condition improvement.The infective tissue and vegetation were aggressively debrided after the infective prosthetic valve removed.Before the new valve was replaced paravalvular tissue must be flushed with diluted iodine solution and large quantities of normal saline.Tricuspid valve repair (TVR)was performed during the same period in 3 cases.Results Hospital death occurred in 8 patients and only 2 patients were recovery in group with medical treatment only.The main reasons of death were infective shock and cardiac failure in 4 patients,cerebral complications(embolism or bleeding)and multiple organs failure in 4 cases.While 6 patients cured after combined medical and surgical management.Follow-up had been carried out in 8 cases for 1.7 to 15 years(mean 5.1 years).Eight years later one patient was re-operated because of severe tricuspid regurgitation and paravalvular leak.There was no PVE recurrence in others.Conclusion Combined medical and surgical management for PVE get a better result than medical treatment alone.Good prognosis lies in timely diagnosis,adequate antibiotics,aggressive debridement of infective tissue and better myocardium protection during operation.

Determination of tetrahydropulmatine content in treatment of winter's diseases in summer path by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a high performance liquid chromatography method for the determination of tetrahydropulmatine content in treatment of winter's diseases in summer path.</p><p><b>METHOD</b>The analysis was performed on the Hypersil ODS2 column (4.6 mm x 250 mm, 5 microm) with methanol and 0.1% phosphoric acid solution (adjusted pH to 6.28 with triethylamine) as mobile phase at 63:37. The flow rate was 1.0 mL x min(-1) and the UV detection wavelength was 280 nm.</p><p><b>RESULT</b>The linear range of tetrahydropulmatine was obtained from the range of 2.08 - 49.92 mg x L(-1) (r = 0.999 8). The average recovery was 97.85% with RSD 2.49%.</p><p><b>CONCLUSION</b>This method is simple, reliable and can be used for the determination of tetrahydropulmatine in this preparation.</p>

Analysis of median-term therapeutic effect of coronary artery bypass grafting in the elderly patients / 中国综合临床

Objective To summarize the clinical experiences and early-to median effect of coronary artery bypass grafting(CABG)in patients aged 70 years or above.Methods From Dec.2006 to Dec.2007,86 patients aged 70 or above underwent CABG,of whom 71 cases underwent off-pump coronary artery bypass grafting(OPCAB),15 cases had CABG with cardiopulmonary bypass(CPB).5 cases underwent mitral valve replacement,3 underwent aortic valve replacement besides CABG,and one case underwent removal of ventricular aneurysm.Their clinical data were retrospectively analyzed.Results There was one case who died of enterobrosis after operation.The average ICU stay time was(77.4±49.6)h,the average postoperative stay time was 15 d,and the average drainage was 530 ml.22 postoperative complications were found.76 cases were followed up for 10-22 months.Follow up effect was better showing cardiac classification I-II and no angina pectoris,myocardial infarction and late death.Conclusion Patients with coronary heart disease in good condition will have not remarkable improvement in mortality as compared with ordinary patients but attention should be paid to their preoperative pulmonary function and angina pectoffs.OPCABG would have encouraging result in reducing the rate of postoperative complications and hospital death,which should be recommended as the first choice for the elderly patients.

Clinical analysis of 1386 patients undergoing off-pump coronary artery bypass grafting / 中国综合临床

Objective To summarize the clinical experiences of 1386 patients undergoing off-pump coronary artery bypass grafting(OPCAB)during Jan 2002 to Mar 2008.Methods OPCAB was done through midsternumtomy under ordinary-temperature anesthesia after radial artery,left breast artery and vena saphena magna were taken.Patients over 70 years old accounted for 32.35%,and the oldest was 99 years old.Emergency and subemergency OPCAB was done in 111 cases,re-do CABG with off-pump technique was done in 52 patients.Results 24 cases died during perioperation,with mortality of 1.73%and all other patients discharged from the hospital.Following operation,the symptoms of angina pectoris were relieved and the life quality was much beaer than that before operation.Conclusion OPCAB is feasible for older patients,who are complicated with other chronic diseases for elder patients.Skillful technique of surgeon,right indications for OPCAB and management of peri-and postoperation are key to success of operation.

Peri-operative management of off-pump coronary artery bypass grafting / 中国综合临床

Objective To retrospectively analyse the clinical data of off-pump coronary artery bypass grafting surgery(OPCAB)in our department and summarize the clinical experience of peri-operative management.Methods From January 2007 to May 2008,253 patients underwent OPCAB.Seventy-six cases with left main disease(including 18 cases of single left main lesion and 58 cases of complex lesion);13 cases had single vessel disease;32 cases had double vessels disease and 190 cases had triple vessels disease.Eighteen cases received emergency OPCAB.All vessel anastomosis were performed with coronary artery stablizer and shunt through median sternotomy.Resuits Three cases died after operation(1.2%).Intra-aortic balloon pump was used in 11 cases.Mechanical ventilation time was 3-168 h[(24.1±22.3)h],and ICU stay time was 1-14 day[(2.8±2.0)d].Postoperative hospital stay time was 8-42 days[(15.6±6.3)d].Conclusion OPCAB is reliable for most bypass cases.With appropilate peri-operative management,a satisfied result will be acceptable.

Correlation between OPN and P21 in non-small cell lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>At present, little is known about the correlation between osteopontin (OPN) and P21 in primary non-small cell lung cancer (NSCLC). The aim of this study is to investigate the expression of OPN and P21 and their correlation in NSCLC.</p><p><b>METHODS</b>The expression of OPN and P21 was detected in 58 NSCLC tissues and 18 pulmonary inflammatory pseudotumor (PIP) as controls by immunohistochemical technique.</p><p><b>RESULTS</b>The positive rate of OPN and P21 in NSCLC was 58.6% and 70.7% respectively, which was significantly higher than those in PIP (11.1% and 16.7%) (P=0.0008, P < 0.0001). There was a positive correlation between OPN and P21 expression (P < 0.0001). Lymph node metastasis was closely related to OPN expression (P=0.0367), but not to P21 expression (P=0.4898).</p><p><b>CONCLUSIONS</b>The abnormal expression of OPN and P21 in NSCLC may act in a synergistic way. Immunopositivity of OPN may suggest a potential risk of metastasis in NSCLC.</p>

Analysis on Safety of Drug Use in 291 Surgical Patients of Our Hospital in 2009 / 中国药房

OBJECTIVE:To probe into the situation and safety of drug use in surgical patients of our hospital and new prob-lems resulted from ppatients controlled analgesia(PCA). METHODS:Medication and the safety of drug used for PCA in 291 surgical patients of orthopedic department and general surgery department of our hospital in 2009 were analyzed. RESULTS:90 catego-ries of drugs were involved and utilization frequency was 2 627 with each person of 9.03. 70.45% of patients received PCA after operation. There were statistical significance in difference of applying anesthetic agents,analgesics and other drugs in different departments(P