Objective To study the apoptosis of human leukemia K562 cells induced by snake venom of Agkistrodon Halys Pallas in Zhejiang Province.Methods IC_ 50 value and cytotoxity of K562 cell were detected by MTT method.Apoptotic cells were dyed by Hoechest 33258.Sub-G1 peak and cell cycle were detected by FCM.Protein expression of Bcl-2 gene was detected by FCM and western-blot method.Results The snake venom of Agkistrodon Halys Pallas in Zhejiang Province inhibited the growth of K562 cells,which appeared dose-dependent.The snake venom induced apoptosis of K562 cells.Meanwhile,protein expression of Bcl-2 was down-regulated.Conclusion Snake venom of Agkistrodon Halys Pallas in Zhejiang could induce apoptosis of human leukemia K562 cells.The mechanism may be related to down-regulation of Bcl-2 gene expression.

Objective To analyze the value of pulse sequential continuous cardiac output(PICCO) in chronic pulmonary heart disease with cardiogenic shock.Methods Sixty-eight patients with chronic pulmonary heart disease complicated with cardiogenic shock were randomly divided into study group(n=36) and control group(n=32).Both groups were treated with conventional therapy,while the control group underwent deep venous puncture to monitor the central venous pressure (CVP) changes.The study group underwent deep venous catheterization+ femoral artery catheterization monitoring + PICCO monitoring.The therapeutic effect and the changes of PICCO index of the two groups were observed.Results The time of administration of vasoactive drugs,hospital stay and mechanical ventilation were significantly lower in the study group than in the control group (P＜0.05).There was no significant difference in mortality rate between the two groups(P＞0.05).After treatment,cardiac output quantity (CI) in study group increased significantly and extravascular lung water index EVLWI and PVPI decreased significantly,the difference between the two groups were statistical significance (P＜0.05),ITBVI had no statistical significant difference betweent two groups(P＞0.05).CI in the survival group was significantly higher than the death group,PVPI and EVLWI were significantly lower than the death group (P＜0.05),and the two groups had no significant difference in ITBVI (P＞0.05).Conclusion PICCO can reflect the hemodynamic status of patients with chronic pulmonary heart disease complicated with cardiac shock,and it has important clinical value for guiding the treatment and prognosis of patients.

Objective To investigate the value of prenatal ultrasound observation of subaortic conical connection in the diagnosis of complete transposition of great arteries(TGA).Methods Echocardiography findings of 22 cases with complete TGA confirmed by autopsy were retrospectively analyzed and summarized.Results All cases were shown subaortic conical connection on two-dimensional echocardiography,the two great arteries were parallel in 18 cases on left ventricular long-axis view,the character of normal right ventricular outflow tract surrounding the aortic root disappeared in 17 cases.There were 13 cases with ventricular septal defect and 2 cases with left ventricular outflow tract obstruction.Aorta and pulmonary artery were shown in right anterior and left posterior position among 20 cases,and in anterior and posterior position in 2 cases.Conclusions Prenatal ultrasound observation of subaortic conical connection is of great value in the diagnosis of complete TGA.

Objecitve To investigate the relationship between the level of pregnancy-assoiated plasma protein-A(PAPP-A) and high-sensitivity C-reactive protein(hs-CRP) in patiente with acute coronary syndrome(ACS) and the role of PAPP-A in predicting the stability of atherosclerotic plaques.Methods A total of 43 patients were selected as ACS group,25 patients as stable angina(SA) and 13 persons as control.ACS group ansisted of 20 patients with unstable angina(UA) and 23 patients with acute myocardial infarction(AMI).The peripheral venous blood samples of the both control and angina groups were drawn on the morning of the following day after hospitalization, and of the AMI group were drawn 24 hours after episode.The concentration of PAPP-A was detected by enzyme-linked immune sorbent assay(ELISA),and the concentration of C-reactive protein(CRP) was detected by scatter turbidimetry method.The concentration of PAPP-A and CRP in each group was compared.Results The plasma hs-CRP and PAPP-A levels in patients with AS increased significantly compared with those in UA and healthy volunteers; PAPP-A was significantly related with hs-CRP(r=0.308);using the method of multiple regression and correlation PAPP-A had linear relationship with hsCRP in all of the cases.Conclusion PAPP-A can play a role in evaluating the clinical stability of patients with coronary disease,and may serve as one of the biomarkers of vulnerable plaques.

Objective To observe the effects of transcutaneous electric acupoint stimulation(TEAS) at zusanli on fatigue after training.Methods 40 soldiers with exercise fatigue after marine training were divided into 2 groups.The soldiers in treatment group were treated with TEAS on Zusanli(ST36).Those in control group recovered naturally.Results The fatigue symptoms were improved,the consistency of the hemoglobin(HGB) were significantly increased,and the levels of lactic acid(LA) were significantly decreased in treatment group(P<0.01),which seemed better than that of the controls(P<0.05).Conclusion TEAS can facilitate the recovery from fatigue after training.

Objective To observe the efficacy of extensive decompression through transforaminal lumbar interbody fusion (TLIF) pathway combined with pedicle screw fixation on treating degenerative lumbar stenosis in the dderly.Methods Seventy-five elderly patients (28 males and 47 females) with degenerative lumbar stenosis were treated with extensive decompression through transforaminal pathway at our hospital from Jan.2007 to Aug.2010.The operation is through the TLIF pathway to resect part of the articular facet,and expose unilaterally the intervertebral vertebral foramen.Decompression of the vertebral canal was conducted by removing the disc.In the end,we performed posterolateral fixation with pedicle screw and placement of bone graft in posterolateral part of the lumbar or did the interbody fusion.JOA scores were obtained before and 1 day after operation and in 3 month follow-up consultation.The intervertebral height and bone fusion were observed by X ray.Results The follow-up period of the 75 patients was 6-36 months with an average of 12 months.There was significant difference (t =20.79,P ＜ 0.05 ;t =25.89,P ＜ 20.05) in JOA score between 3 month follow-up (21.08 ± 3.60) and preoperation (10.91 ± 2.23),between 1 d follow-up (22.72 ± 3.26) and preoperation (10.91 ±2.23),respectively.The rate of improvement was (88.6 ± 10.8)％,with 98％ of excellent or good in 3-month follow-up.Lumbar plane films showed neither instability or internal fixation loosening,breakage or distortion in follow-up consultation.There were 2 cases in whom Cage dislocation occurred without any neurological symptoms.Conclusion Extensive decompression through TLIF pathway combined with pedicle screw fixation is an efficacious method of treating degenerative lumbar stenosis in elderly patients.This method can retain the structure of lumbar posterior complex,reduce the risk of low back pain.It is a safe choice for treatment of degenerative lumbar stenosis in the elderly.

Objective To investigate the effects of 6-minutes of walking exercise (6-MWE) on the exercise tolerance and left ventricular diastolic function (LVDF) of heart failure patients with a normal ejection fraction (HFNEFs).Methods Ninety grade Ⅱ or Ⅲ HFNEFs of the New York heart association (NYHA) were randomly divided into an exercise training group and a control group with 45 cases in each.The control group was treated with routine drugs.The exercise training group was treated with the same drugs plus 6-MWE.Before and after the sixmonth period of treatment,plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) levels were determined,each subject's left atrial volume index (LAVI) was measured with a color ultrasonic cardiogram (UCG),and their 6-minute walk distance (6-MWD) was measured.Results Plasma NT-proBNP levels and 6-MWD improved significantly comparing with before treatment in both groups.The average 6-MWD,LAVI and plasma NT-proBNP level all improved significantly more in the experimental group.Conclusion 6-MWE can significantly improve the exercise tolerance and LVDF of HFNEFs,and improve their quality of life.Walking can be helpful in delaying the development of HFNEF.

10.3969/j.issn.1000-3606.2013.06.010

Objective To explore the correlation between brainstem auditory evoked potential (BAEP) combined with mesencephalic morphology and the prognosis of coma patients with severe craniocerebral injury.Methods Forty-seven coma patients with severe craniocerebral injury were examined with BAEP and moaitored dynamically in early period of coma,and all these patients got head CT scans and the ratio of anteroposterior diameter and transverse diameter of mesencephalon were measured at the same time.Results There was a significantly negative relationship between the first BAEP classification and GOS score at the end of the follow-up (r =-0.755,P =0.000 ).Among all of the patients,there were 27 patients with the ratio of anteroposterior diameter and tansverse diameter of mesencephalon from 0.9 to 1.1,19 patients (70.4％,19/27) with good prognosis;20 patients with the ratio ＞1.1 or ＜ 0.9,6 patients (30.0％,6/20 ) with good prognosis.The rate of good prognosis between the two had significant difference ( x2 =7.521,P =0.006).The sensitivity and specificity were 73.7％(14/19),82.6％ (19/23) at the first BAEP combined with mesencephalic morphology,88.2％ (15/17),100.0％ (21/21) at the second BAEP combined with mesencephalic morphology.Conclusion BAEP combined with mesencephalic morphology can evaluate effectively the prognosis of coma patients with severe craniocerebral injury.

BACKGROUND:There is a linker between Hsp65 and Esat6 coding a segment of water repellent polypeptide.It is very gentle and easy to be folded,which is profitable to translate the keno-folding correctly between the two proteins and to make the space structure of fusional proteins consistent with those two native ones.Then,a correct structure is formed and the immunogenicity of the fusional proteins is improved.OBJECTIVE:This study was to clone the fusional genes Hsp65-Esat6 from mycobacterium tuberculosis(MTB)H37Rv, then to construct into a eukaryotic expressing vector which contains an enhanced green fluorescent protein(EGFP) reporter gene,and finally to identify the expression of Hsp65 protein and Esat6 protein by IHC methods.DESIGN:Single sample experiment.SETTING:Department of Microbiology ＆ Immunology,Medical College of Jinan University.MATERIALS:Plasmid pVAE was donated by Professor Cao from South China University of Technology.MTB H37Rv,E.coil DH5 α,expressing plasmid pEGFP-C1 and Hela cells were donated by Doctor Hu Ping.METHODS:The experiment was conducted at the Department of Microbiology ＆ Immunology and the Medical Experimental Center of Jinan University between September 2005 and June 2006.The whole-genome was extracted from MTB H37Rv by molecular cloning technique,and used it as template to amplify Hsp65 (no terminator) gene by polyrnerase chain reaction (PCR),to recombine it with pEGFP-C1 vector after purification to construct pEGHsp65(no terminator)recombination vector.pVAE vector was used as template to amplify Linker-Esat6 gene(with terminator)by PCR, and then to recombine it with pEGHSP65(no terminator)to construct an eukaryotic expressing vector pEGHsp65-Esat6 with the fusional genes Hsp65-Esat6 inside.Finally,genes Hsp65-Esat was checked by molecule biology methods such as PCR, restriction endonuclease and DNA sequencing.Hela cells were transfected with pEGHsp65-Esat6 and the expression of EGFP and the efficiency of transfection were observed.The expressions of Hsp65 protein and Esat6 protein were detected by IHC methods.MAIN OUTCOME MEASURES:①The size of pEGHsp65-Esat6.②The DNA sequencing result of the pEGFP-C1.③The expression of EGFP in the transfected Hela cells.④The IHC results of Hsp65 protein and Esat6 protein in Hela cells.RESULTS:①The size of pEGFP-C1 was 4.7 kb,that of pEGHsp65 was 6.4 kb,and that of pEGHsp65-Esat6 was 6.7 kb.There were differences between their speeds In agarose electrophoresis.②The results showed that it was the same as reported Hsp65 sequence and Esat6 sequence of MTB H37Rv.③After transfecting pEGHsp65-Esat6 for 24 hours,EGFP was found in 30% of Hela through Laser scanning confocal microscope. But there was no EGFP in non-transfected Hela.④Hsp65 protein and Esat6 protein with biological activities were detected in transfected Hela cells by IHC methods.CONCLUSION: Using Hsp65 and Esat6 as immunogens,we have successfully cloned and constructed a eukaryotic expressing vector which contain fusionaI genes Hsp65-Esat6 of MTB H37Rv and EGFP.