Objective To realize the construction and expression of human dsFv genes to rabies virus. Methods The genes of dsFv VH and VL were amplified by PCR-based point mutagenesis strategy. Then the genes were cloned into plasmid pET-22b (+). E. coli BL21(DE3) was transformed with the recombinant expression plasmid and the protein was induced in Luris-Bertani medium by addition of IPTG. The inclusion body proteins of VH and VL were denaturalized by GuHCl, and then re-naturalized in refolding solution to form dsFv fragments. Afterwards, we evaluated the antigen-binding activity of the dsFv and its stability as compared with its originative scFv. Results The fragment genes of dsFv to rabies virus were constructed successfully by PCR-based point mutagenesis strategy. Sequence analysis proved that cysteines were introduced into the position 44 aa of VH and position 100 aa of VL. The dsFv VH and VL genes were expressed in PET22b(+)/BL21(DE3). The VH and VL protein folded into the active dsFv antibody fragments which showed specific binding capability to rabies virus. Conclusion We succeeded in achieving the stabilization of the human scFv to rabies virus and obtaining the active human dsFv antibody fragments. This established a solid basis for further study of the biological activaty and clinical application of dsFv to rabies virus.

Objective To assess the changes of left ventricular function after off-pump coronary artery bypass (OPCAB) using Tei index. Methods Fourty-six patients of coronary artery disease, who had accepted OPCAB, were enrolled. Left ventricular ejection fraction(LVEF), E/A ratio at mitrial orifice(E/A), E deceleration time(EDT), isovolumic relaxation time(IVRT), ratio of systolic wave to diastolic wave of pulmonary vein flow(S/D) and Tei index were measured by transthoracic echocardiography before, immediate, 1 week, 1 month, 3 months and 6 months after OPCAB. Results Tei index decreased gradually as time went on after OPCAB and the differences were significant between different times(P

Objective To evaluate the impact of the off -pump coronary artery bypass (OPCAB) on left ventricular(LV) function with transesophageal echocardiography(TEE). Methods Blood flow velocities of pulmonary vein (PV) and mitral valve (MV) were performed in 21 patients with coronary heart disease and myocardial infarction two weeks pre-OPCAB, and the left ventricular SV, CO and LVEF were measured by biplane Simpson′s method, the data were compared with the results observed one month post-OPCAB. Results The peak flow velocities of PV had significant difference between pre- and post-OPCAB(P

Objective To summarize the experiences in clinical diagnosis and surgical treatment of chest cancers invading the superior vena cava (SVC). Method Retrospective study of 12 cases with chest cancers invading the SVC, SVC and innominate vein were reconstructed with prosthesis in 3 cases, the side wall of the SVC was excised and repaired in 9 cases. Results Complete resection was performed in 10cases,2 cases had incomplete resection. There was no death associated with operation in these patients. All patients were followed up for 1 - 7 years after operation,and 3 cases were lost to follow up,5 of remaining 9cases were dead because of relapse and metastasis, their median survival time was 8 months,other 4 cases lived, their median survival time was 42 months. Conclusions Patients with chest cancers invading the SVC should have surgical treatment of not only tumor resection but also the vessel reconstruction,the prognosis would be better than without surgical treatment. These patients have a high quality of life and live with long-term survival.

OBJECTIVE:The effects of Astragulus injection on cardiac function in myocardial infarction convalescents were observed.METHODS:42 cases were randomly divided into two groups(therapeutic group and control group).21 cases in therapeutic group were treated with routine treatment and 21 cases in control group were treated with Astragulus injection on the basis of routine treatment for three weeks.The ejection fraction(EF),early diastolic peak velocity of blood flow(E),late diastolic peak velocity of blood flow(A) and A/E of all patients were measured with pulse ultrasonic Doppler cardiography before and after the treatment.RESULTS:After the treatment,the EF,E and A/E were improved (P

Objective To investigate the relapse factors and its control strategy for peptic ulcer patients .Methods A total of 160 cases of peptic ulcer patients who visited our hospital from Jan 2011 to Aug 2013 was retrospectively analyzed .The recurred pa-tients were assigned as the experimental group , and the others as the control group .The relapse factors for peptic ulcer were observed . Results Among 160 peptic ulcers , there were 46 cases with recurrence with a relapse rate of 28.8%.Single factor logistic regression analysis showed that smoking, drinking, the spring and autumn season, h.pylori (Hp) infection, non-steroid anti-inflammatory drugs (NSAIDs), mental factor, and male were the risk factors for recurrence of peptic ulcer .Further multivariable logistic regression analy-sis showed that smoking, drinking, mental factors, history taking NSAIDs, and Hp infection were also the risk factors for recurrence of peptic ulcer .Conclusions Smoking and alcoholism history , meatal factors , history of taking NSAIDs , and Hp infection were the in-dependent risk factors of recurrence of peptic ulcer , intervention of these risk factor might reduce ulcer recurrence .

Objective To evaluate the methods of standard patient (SP)combined with case-based learning (CBL ) in humanistic quality cultivation for interns in the department of thoracic surgery. Methods Medical interns in the department of thoracic surgery were divided randomly into experimental(n=30)and control group(n=30)respectively. In experimental group,SP combined with CBL was used while in control group traditional teaching was used. At the end of the study,teaching effect was assessed by the final examination and questionnaires and the data were processed by t test andX2 test. P<0.05 signifies statistical differences. Results There was no significant difference in test scores between the two groups (P>0.05). Scores of history-taking,medical check-up,paper writing quality in the final examination in experimental group were significantly higher than those in control group(P<0.01). Satisfaction degrees of doctor-patient communication,responsibility and service attitude in experimental group were better than those of control group(X2=16.484,10.335,20.376,32.851,P<0.01). Conclusions SP combined with CBL teaching method can stimulate the students' interests in thoracic surgery practice,improve the doctor-patient relationship and strengthen the ability of human-istic concern. It is a better clinical thoracic practice teaching method and it is beneficial to cultivating medical students with humanistic concern.

[Objective] To summarize image manifestations in intestinal tuberculosis (ITB) examined by multi-slice computed tomography enteroclysis (MSCT) and evaluate the diagnostic value of MSCTE in ITB,[Methods] The imaging findings were retrospectively analyzed in 15 cases of ITB that were confirmed by endoscopy or postoperative pathologic examination,including the location,number,shape,edge,surrounding tissue alterations of ITB and other associated changes in the peritoneum,mesentery and solid abdominal organs,and compared with endoscopy and pathology.[Results] In 15 patients,ileocecum was involved in 13 cases (87％),4 cases (27％) showed multi-segmental symmetric intestinal mural thickening,9 patients (60％) showed solid masses,1 case (7％) showed multi-segmental symmetric intestinal mural thickening and solid masses,1 case (7％) showed homogenous enhanced masses and perforation,12 cases (86％) showed enlarged lymph nodes (LNs) with rim enhancement,and 2 cases (13％) showed the comb sign of enhanced mesenteric vessels.Based on enhancement pattern of MSCTE,ITB was divided into three types:homogenous enhancement type (n=5);target sign type (n=2);caseous necrosis type (n=2).[Conclusion] The imaging features of ITB are diverse.MSCTE can clearly display the shape of intestinal mucosa,the alterations of intestinal wall and the relationship between lesion and adjacent tissues,which provides valuable information for the clinic diagnosis of ITB.

AIM: To obtain a vaccine with sLAG-3 as immunoadjuvant and investigate its biologic activity in order to establish the safe and effective way for asthma as one of the specific immunotherapy.METHODS: The coding sequence of LAG-3 was amplified by polymerase chain reaction,the expression vector pcDNA-sLAG-3-Ig was constructed by inserting the PCR products of sLAG-3 and Fc sequence of IgG.With electroporation transfection,pcDNA-sLAG-3-Ig was transfected into COS-7 cells and its biologic activity was investigated by Western blotting analysis.RESULTS: By temperature induction,the LAG-3-Ig was highly expressed in E.coli DH5?.LAG-3-Ig fusion protein was observed by SDS-PAGE and Western blotting,the results showed that the LAG-3-Ig protein was an antagonist of the IL-4-induced synthesis of IgE in B cells.CONCLUSION: A new vaccine with sLAG-3 as immunoadjuvant was obtained.It could inhibit synthesis of IgE in B cells.Thus,LAG-3-Ig would be hopeful to establish the safe and effective way for asthma as one of the specific immunotherapy.

BACKGROUND: Recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) could stimulate the proliferation of fibroblasts, keratinocyte and skin mucosae cells to different degrees.OBJECTIVE: To observe the effect of rhGM-CSF on the healing of drug exosmose induced skin ulcers.DESIGN: A randomized and controlled animal experiment.SETTING: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out at the Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University of Chinese PLA from June to November 2004. Totally 20 male Kunming white mice, with body mass of 18 to 24 g, were chosen.METHODS: Prepared skin ulcers animal models were randomly divided into control group and treated group with 10 white mice in each group.Mice in the control group were given 1mg phentolamine ,20 mg lidocaine and 1mg dexamethasone diluted by normal saline to 0.5 mL ,then sealed up , once a day for 7 days; 25 μg rhGM-CSF was diluted by normal saline to 0.5 mL , then the solution was injected into the periphery of ulcers of mice in treated group , once every other day, for 7 days. Healing time and histological change of skin tissue at ulcer were observed.MAIN OUTCOME MEASURES: To observe the effect of rhGM-CSF on the healing time of drug exosmose induced skin ulcer and anabrosis and histological changeRESULTS: Totally 20 mice entered the stage of result analysis. ①Healing time: the healing time of ulcer and erosion was significantly longer in control group than in treated group [(20-24,8-12)d,t=2.264,P=0.01];②Histological observation: hyperplasia of granulation tissue was not obviously on 7 days after treatment in control group; Hyperplasia of granulation tissue appeared and the newly born blood vessel was abundant on 7 days after treatment in the treated group.CONCLUSION: rhGM-CSF can promote the wound healing of drug induced anabrosis and ulcer.