AIM: To study the recombinant plasmid pDL121 and its expression product in E.coli. METHODS: pDL121 was analysed by using 6 different restriction endonucleases and dot blotting, and the isolated 23 kDa protein band was cut and injected twice into rabbits to raise anti - 23 kDa serum. RESULTS: The restriction map of pDL121 was quite different from other leptospiral genes reported and digoxin labeled recombinant DNA probe of pDL121 could detect the pathogenic leptospires, whereas, not the nonpathogenic leptospires. The anti - 23 kDa serum could recognize the sonicated antigen of L. interrogans serovar lai strain 017 and 23 kDa protein expressed in pDL121 and the titer of the antiserum were very high, approximately 1/12 800. Injection of the E. coli lysate of pDL121 with Freund's adjuvant into guinea pigs resulted in some protection of the animal against the challenge with strain 017. CONCLUSION: It indicated that 23 kDa protein had good imunogenicity and could serve as a candidate for protective antigen of L. interrogans and the inserted fragment of pDL121 could be a new gene .

Siderophores were produced and secreted with marine microorganisms as the highly specific iron chelators. A method of liquid chromatography-tandem mass spectrometric detection coupled with solid phase extraction pretreatment was developed for the determination of siderophores in seawater. The samples were filtered through a 0. 22 μm membrane, extracted with an ENVI-18 cartridge and then eluted with methanol. The separation of the analytes was performed on a reversed phase SB-C18 column with a gradient elution program by using 0. 1% ( V/V) formic acid and methanol as the mobile phases. Qualitative analysis was performed in multiple-reaction monitoring mode. Good linearity (R2>0. 99) was obtained for Pyoverdines-Fe, Ferrichrome, Ferrioxamine E at the concentrations of 0. 001-3. 00 μg/mL, 0. 005-15. 00 μg/mL, 0. 001-3. 00 μg/mL, respectively. The instrumental detection limits and limits of quantification for the three analytes were 0. 08, 1. 76 and 1. 36 ng/mL; 0. 27, 5. 87 and 4. 53 ng/mL, respectively. The relative standard deviations were lower than 12%, while the recoveries were 12. 1%-18. 6% for Pyoverdines-Fe, 82. 0%-97. 7% for Ferrichrome, and 70. 0%-98. 3% for Ferrioxamine E.

Objective To investigate the high risk factors,pathology and clinical features in fatal pulmonary embolism in the elderly patients with malignant tumor,and to analyze the characters of the embolus and provide experimental data for clinical prevention and treatment offatal pulmonary embolism.Methods Autopsy and clinical data of 19 malignancy cases with fatal pulmonary embolism admitted to Beijing Hospital from 1975 to 2006 were retrospectively analyzed.Results 33.9％ of total pulmonary embolism were from malignant diseases (19/56).Saddle and massive pulmonary embolism were in 84.2％ of total 19 cases,and micro-embolism cases were in only 15.8％.The 84.2％ of embolisms were from pulmonary thromboembolism,15.8％ from tumor emboli,and 5.26％ from fungi emboli.Pulmonary adenocarcinoma was the most common (36.5％),the second was pancreatic cancer (15.8％).In all the clinical symptoms,78.9％ of symptoms were dyspnea,15.8％ were syncope.Tachycardia and cyanosis were the common physical signs.Conclusions Malignant tumor is an important risk factor for pulmonary embolism in elderly patients.There are various kinds of emboli in pulmonary embolism in elderly patients with malignant tumor.We should comprehensively consider the causes of pulmonary embolism and give the reasonable and effective treatment to the patients.

AIM: To study the recombinant plasmid pDL121 and its expression product in E.coli. METHODS: pDL121 was analysed by using 6 different restriction endonucleases and dot blotting, and the isolated 23 kDa protein band was cut and injected twice into rabbits to raise anti-23 kDa serum. RESULTS: The restriction map of pDL121 was quite different from other leptospiral genes reported and digoxin labeled recombinant DNA probe of pDL121 could detect the pathogenic leptospires, whereas, not the nonpathogenic leptospires. The anti-23 kDa serum could recognize the sonicated antigen of L.interrogans serovar lai strain 017 and 23 kDa protein expressed in pDL121 and the titer of the antiserum were very high, approximately 1/12 800. Injection of the E.coli lysate of pDL121 with Freund's adjuvant into guinea pigs resulted in some protection of the animal against the challenge with strain 017. CONCLUSION: It indicated that 23 kDa protein had good imunogenicity and could serve as a candidate for protective antigen of L.interrogans and the inserted fragment of pDL121 could be a new gene .

Objective:To increase the understanding of neuroleptic malignant syndrome, rhabdomyolysis and acute renal injury in advanced-aged patients with Parkinson's disease after abdominal surgery.Methods:We report a case of malignant syndrome, rhabdomyolysis and acute renal injury in an 85-year-old patient with Parkinson's disease after abdominal surgery in our department.The diagnosis and successful treatment experience were summarized, and a literature review was conducted.Results:The body temperature was as high as 40.5℃ in this patient, accompanied by stiffness, sustained involuntary shaking, increased muscle tone, serum creatine kinase at 104 615 U/L, tachycardia, low blood pressure, accelerated breathing rate, disturbance of consciousness, excessive sweating and other clinical manifestations, which met the diagnostic criteria for neuroleptic malignant syndrome.The patient had complications including concurrent rhabdomyolysis, acute renal injury and shock.The emergency was resolved after an early diagnosis and proactive treatment.Conclusions:If patients with Parkinson's disease have a high fever with rigidity or sudden aggravation within a short period of time after medication, the possibility of neuroleptic malignant syndrome should be considered and the causes should be screened.

Objective To study the diagnosis and treatment of Mycobacterium avium complex lung disease with severe renal insufficiency,and to provide a basis for improving the clinical diagnosis and treatment of MAC lung disease with severe renal insufficiency.Methods Clinical data of an elderly patient with MAC lung disease confirmed by induced sputum culture and complicated with chronic kidney disease (CKD 4) was reported.The related literatures at home and abroad were reviewed.Results A male patient aged 80 years with poor nutrition was suffering from progressive weight loss,exhaustion and night sweats.His weight was only 43-44 Kg.Chest CT showed that multiple small nodules,small spot pieces of shadow and ground glass shadows in bilateral lung tissues.PET/CT indicated that spot pieces and nodules with metabolic activity in high performance.Bronchoalveolar lavage fluid (BALF) inspection was negative.MAC was identified by induced sputum culture through high permeability brine induced sputum.It was difficult to choose drugs for the treatment of MAC lung disease due to his poor kidney function [GFR of left kidney:9.0 ml /min,GFR of right kidney:18.8 ml/min].Conventional anti-mycobacterium drugs showed a low to high resistance to MAC.Moxifloxacin was discontinued for renal insufficiency.His condition was stable after choosing trimodality therapy including azithromycin 0.25 g/d tiw,rifapentine 0.3 g/d biw,ethambutol 0.375 g /d tiw and the joining immunotherapy.Conclusions PET/ CT is not useful in identifying tuberculosis and MAC lung disease.The treatment of MAC lung disease is difficult in elderly patients with severe renal insufficiency and low weight.Individual therapy combined with immunotherapy and improving the nutrition state is a good choice for the treatment of MAC lung disease in elderly patients with severe renal insufficiency.The anti-MAC drugs should be carefully selected and the adverse reactions should be closely observed in order to obtain the satisfactory clinical results.

Objective To evaluate the therapeutic effects and complications of modified facial mask for non-invasive ventilation (NIV) in elderly patients with respiratory failure.Methods A total of 132 elderly patients(107 males and 25 female,aged 78.5±8.6 years) treated with NIV from February 2008 to May 2011 were randomized into two groups:modified facial mask(group A,n=68,56 males and 12 females,aged 78.8±22.2 years) and control facial mask(group B,n=64,64 males and 13 females,aged 76.6±20.4 years).Duration of NIV,time in RICU(respiratory intensive care unit),length of hospital stay,risk for hospital acquired pneumonia (HAP),risk for invasive ventilation,cure rates,in-hospital mortality,NIV failure rate and cost were compared between the two groups.The complications of NIV,such as oropharyngeal dryness,skin damage of face and nose,abdominal bloating,gas leakage from mask were also compared between the two groups.Results Compared with group B,duration of NIV(12.2±2.3 d vs.18.4±3.6d),time in RICU(7.3±3.2d vs.14.6t5.4d),length of in hospital stay(16.6±4.2d vs.28.2±6.2)d,and cost(2.23±0.12 ten thousand yuan vs.4.23± 0.24 ten thousand yua) in group A were significantly decreased(t=9.72,14.91,13.08,10.81 respectively,all P＜0.05).The risk for invasive ventilation [2.9％ (2 cases) vs.43.8％(28 cases)],NIV failure rate [5.9％ (4 cases) vs.12.5％ (28 cases)] were also decreased in group A compared with group B(x2 =31.26,25.74,both P＜0.05).Compared with group B,The complications of NIV such as skin damage of face and nose[4.4％ (3 cases) vs.37.5％ (24 cases)],abdominal bloating [2.9％ (2 cases) vs.28.1％ (18 cases)],gas leakage from mask [8.8 ％ (6 cases)vs.50％(32 cases)] in group A were significantly decreased(x2 =31.26,25.74,all P＜0.05).Conclusions Modified facial mask for NIV is effective in the treatment of elderly patients with respiratory failure.The complications and in-hospital mortality are reduced with the application of modified facial mask for NIV and it is highly tolerated by patients.Modified facial mask for NIV is the first choice in the treatment in elderly patients with respiratory failure.

Objective To obtain a novel tool-enzyme for genetic engineering with good solubility,strong specificity of enzyme digestion and maintaining the enzyme activity at low temperature by using E.coli expression system to express self-processed re-combinant MBP-HRV 3C fusion protease.Methods The cDNA encoding HRV 3C protease was cloned into pRSF-Duet vector.The recombinant plasmid was transferred into E.coli BL21 (DE3)for expression.HRV 3C protease was obtained through Nichol col-umn affinity purification.The cleavage activity of HRV 3C protease was determined by in vivo experiment.Results HRV 3C prote-ase was highly expressed in E.coli expression system,and the obtained HRV 3C protease could recognize and digest HRV 3C site. Conclusion A novel tool-enzyme for genetic engineering is obtained.

Objective:To investigate risk factors for failure of non-invasive ventilation(NIV)in elderly patients with acute hypercapnic respiratory failure(AHRF).Methods:Retrospective analysis was performed on the data of 100 elderly patients hospitalized for NIV treatment for AHRF at Beijing Hospital between August 2015 and August 2017.The patients were divided into an NIV failure group and an NIV success group according to patient treatment results.Logistic regression was used to analyze the related factors of NIV treatment failure in elderly patients with AHRF.Results:Of the 100 patients, there were 53 men(53.0%), with an age of(79.4±6.4)years, and 43(43.0%)failed with NIV treatment.Before NIV treatment, concurrent severe pneumonia( OR=5.109, 95% CI: 1.203-21.702, P=0.027), chest X-ray showing lesion infiltration≥3 quadrants( OR=5.903, 95% CI: 1.514-23.013, P=0.011)and a simplified acute physiology score Ⅱ(SAPS Ⅱ)≥34 points( OR=7.046, 95% CI: 2.164-22.944, P=0.001)were independent risk factors for failure of NIV treatment in elderly patients with AHRF. Conclusions:Elderly patients with AHRF face the risk of failure with NIV therapy.Before NIV treatment, concurrent severe pneumonia, chest X-ray showing lesion infiltration ≥3 quadrants and SAPS Ⅱ≥34 points are independent risk factors for failure of NIV treatment in elderly patients with AHRF.Evaluation before NIV therapy is important to prevent NIV treatment failure.

Objective To evaluate the effects,safety and economic cost of long-term home noninvasive ventilation (NIV) therapy in elderly patients with chronic hypercapnic respiratory failure.Methods A total of 128 elderly patients with chronic hypercapnic respiratory failure were randomly assigned to two groups:the NIV group (n=66) with conventional therapy in addition to long-term home NIV therapy,and the control group (n=62) with conventional therapy alone.Compared were parameters before and after two year follow up,which included dyspnea grade,scale for accessory muscle use,scoring for emotional disorders,mean pulmonary pressure (mPAP) by electrocardiography,arterial blood gas,the times of pulmonary infection and hospitalization rates,the duration of hospitalization invasive ventilation,the duration of in RICU and in hospital stay,tracheal intubation rates and mortality.The medical cost was calculated.Results After two years,the differences in the dyspnea grade,scale for accessory muscle use,anxiety scores,depression scores,mPAP,arterial PaCO2 and PaO2,hospitalization rates,the times of pulmonary infection,the days of hospitalization for exacerbation in the home NIV group [2.2± 0.2,2.4 ± 0.3,4 ± 1,5.3 ± 1.2,(36.6±5.2)mm Hg,(50.2±4.5)mm Hg,(63.5±4.2)mm Hg,(1.3±0.2) times/year,(2.4±0.2) times/year,(15.8 ± 4.4) days/times] were statistically significant compared to the control group [4.1±0.2,4.9±0.5,12±3,11.3±1.6,(45.2±5.2)mm Hg,(67.3±4.5) mm Hg,(48.3±4.3)mm Hg,(5.4±0.4)times/year,(8.9 ±0.3) times/year,(38.5± 6.3) days/times] (all P＜0.01).The duration of invasive ventilation,the days in RICU and in hospital stay,tracheal intubation rates on admission to the hospital were significantly decreased in the home NIV group [(8.2 ± 2.2)days,(9.6±3.1) days,(15.8±4.4) days,(2±0.2) times/two years],as compared with the control group [(15.8±3.4) days,(18.6±4.4)days,(38.5±6.3)days,(8.0±0.8) times/two years].The mortality was decreased significantly in the home NIV group (3.0 ％)compared with the control group (29.0％) (P＜0.05).The medical cost in two years was significant lower in the home NIV group [(6.4 ± 0.5) thousand yuan],compared with the control group (18.4 ±0.6) thousand yuan (P＜0.01).Conclusions Long-term home NIV therapy in patients with chronic hypercapinc respiratory failure is effective,safe and can decrease the mortality and medical cost.