A Strategy was established for construction of repertoire antibody library with affinity chromatography purifying antigen, antigen immunizing human lymphocytes, RT-PCR and phage display technology. The colorectal carcinoma-associated antigen CA-Hb3 was purified with affinity column and analysed with SDS-PAGE and Western-blot, then applied for immunizing peripheral blood lymphocytes (PBL) in vitro. The PBL were isolated from ten patients with colorectal carcinoma and cultured with interleukin 2 and pokeweed mitogen, lymphoblast-like cells occurred and colonies formed after immunization. Three VH-CHl(?) and five VL-CL(?,?) genes were amplified from their total RNA and mRNA with RT-PCR. Three VH (?) and 8 VL (?,?) genes were reamplified and randomly combined to construct 24 single-chain variable fragments (ScFv) genes through (Gly_(4)Ser)_(3) linker. ScFv genes digested with Sfi I were cloned into fUSE 5 vector and transformed into MC1061 with electroporation. Repertoire antibody library was obtained with 10~(6) tetracycline-resistant colonies, in which the percentage of ScFv inserts was 85 % . This strategy might be used for humanizing mouse-original monoclonal antibody.

Spinal cord injury (SCI) is a common traumatic disease. SCI causes nutritional and metabolic abnormalities including malnu-trition and overnutrition. This article reviewed nutritional metabolism assessments, the changes and reasons after SCI. Spinal Nutrition Screening Tool (SNST) and Malnutrition Universal Screening Tool (MUST) are commonly used to evaluate malnutrition in adult. Screening Tool for the Assessment of Malnutrition in Paediatrics (STAMP) is used to evaluate malnutrition in child. Body mass index, waist circumfer-ence and dual-energy X-ray are used to evaluate overnutrition. SCI causes increasement of total caloric intake, disturbance of carbohydrate metabolism, disturbance of fat and protein metabolism, increasement of metabolic syndrome incidence, high risk of coronary heart disease, and abnormal bone metabolism. Individual diet program should be provided to SCI patients for balanced nutrition. Periodic assessments are also necessary.

Objective To summarize the mid-term result and clinical experience of individual treatment for Stanford type B aortic dissection.Method 213 patients (172 male,41 female) with Stanford type B aortic dissection were admitted between Apr 2008 and Jan 2013.Age ranged from 23 to 83 averaging 54 ± 9.63 patients were treated conservatively and 47 patients underwent open surgery,103 patients by endovascular repair.Result Patients in conservative group were older than those in open surgery group.Preoperative waiting time was the longest in open surgery group.Concomitant procedures adopted in open surgery group included replacement of ascending aorta and aortic arch in 3 patients and fenestration of stented elephant trunk in 19 patients; 31 patients received bypass graft of cervical vessels during endovascular repair.Mortality was 11.1％,4.3％,3.9％ in conservative group,open surgery group and endovascular group,respectively.Compared with open surgery,endovascular repair had a lower incidence of perioperative complications (P ＜ 0.05).186 patients were followed-up with a mean follow-up duration of (26± 17) months.There were no difference in mortality and complications between the three groups.Conclusions In Stanford B aortic dissection,individualized therapy based on patient's condition achieves a favorable mid-term result with low mortality and complication rate.

Objective To observe the influence of spine kyphotic deformity on postmenopausal osteoporosis patients' pulmonary function and physical activity,and then provide a guide for the rehabilitation training of patients with osteoporosis and a kyphotic spine.Methods Sixty postmenopausal women with osteoporosis were recruited from the osteoporosis outpatients of the Third Hospital of Peking University.Half of them suffered from spine kyphosis (the spine kyphosis group) and the other half did not (the non-spine kyphosis group).There was no significant difference in the average ages and body mass indexes of the two groups.The research applied Cobb's angle and wall-occiput distance (WOD) to assess kyphosis severity,a visual analogue scale (VAS) to evaluate the patient's pain level,plus pulmonary function assessments and thoracic expansion assessments in both groups simultaneously,and the 6-minute walking test (6-MWT) to assess the patients' physical activity.Results The spine kyphosis group said they experienced significantly more pain than the non-kyphosis group,and they had worse pulmonary function and poorer physical activity.These differences were statistically significant.Pulmonary function was significantly correlated with the severity of spine kyphosis,back ache,thoracic expansion as well as physical activity.Conclusion Spine kyphosis deformity can aggravate back ache among postmenopausal osteoporosis patients,and affect their pulmonary function and physical activity.When planning clinical exercise training for such patients,physicians should pay attention to respiratory function training so as to improve the patients' pulmonary function,physical activity and endurance as much as possible,and thus improve their quality of life.

Objective To investigate the expression of Skp2 and C-myc in hepatocellular carcinoma and its relationship with clinicopathologic characteristics.Methods The expression of skp2 and C-myc was evaluated in hepatocellular carcinoma tissues of 48 patients,liver cirrhosis tissues of 20 cases,and normal liver tissues of 16 patients by immunohistochemistry(PV9000 two steps methods).Results The expression rate of skp2(33.3%) in hepatocellular carcinoma tissues was significantly higher than that in liver cirrhosis tissues((negative) expression)(P=0.008)and normal liver tissues(negative expression)(P=0.020).The(expression) of skp2 was correlated with histological differentiation and metastasis(P﹤0.001 and P=(0.017)),but not with tumour size(P=0.058),and it was not expressed in highly differentiated(hepatocellular) carcinomas.The expression rate of C-myc protein in hepatocellular carcinoma tissues((58.3)%) was significantly higher than that in liver cirrhosis tissues(15%)(P=0.001) and normal tissues((negativ)e expression)(P

Objective To investigate the cardioprotective effects of 3-nitropropionic acid (3-NPA) pretreatment against ischemia-reperfusion (I/R) injury. Methods Sixteen male Wistar rats weighing 200-250 g were randomly divided into 2 groups ( n = 8 each):(1) 3-NPA group received intraperitoneal 3-NPA 4 mg?kg-1 24 h before the animals were sacrificed and (2) control group received normal saline instead of 3-NPA. The animals were sacrificed and the hearts were immediately removed and mounted on Langendorff apparatus and perfused with K-H solution saturated with 95% O2 and 5% CO2 at 37℃ . After being perfused for 30 min the hearts were subjected to 30 min global ischemia by suspension of perfusion followed by 60 min reperfusion. The HR, left ventricular developed pressure (LVDP) and ? dp/dtmaxd were recorded before ischemia and at 30 and 60 min of reperfusion. Coronary effluent was collected at 15 min of reperfusion for determination of CK and LDH activity. At the end of 60 min reperfusion the hearts were removed for determination of myocardial MDA content and SOD activity.Results LVDP and ? dp/dtmax recovered significantly better in 3-NPA group than in control group. The myocardial MDA content, CK and LDH release were significantly lower in 3-NPA group than in control group. The myocardial SOD activity was significantly higher in 3-NPA group than in control group. Conclusion Chemical preconditioning with 3-NPA protects the heart from I/R injury.

Objective To improve the traditional percutaneous cholecystolithotripsy in order to elevate the success rate and to discuss its indications. Methods With selfmade apparatus, we modified the open procedure which is otherwise impossible in removing the stones. We have followed 120 patients for 3 to 36 months to look into the recurrence of cholelithiasis. Results We have succeeded in 287 operations in a total of 291 (98 6%) cases. Among which 99 cases (34 5%) received modified open procedures, and the postoperative course was all uneventful. 120 patients were followed up postoperatively from 3 to 36 months and the stone recurrence rate was 13 3% (16 cases). Statistical analysis showed that the stone recurrence rate in patients with 3 stones and up, or poor gallbladder function was significantly higher than those with less than 3 stones and or good gallbladder function.Conclusions The percutaneous cholecystolithotripsy with improved procedures can effectively increase the success rate of operation. According to our result the indications should be restricted to those with less than 3 stones and or with good gallbladder functions.

Objective: To observe the effect of on modified St.Thomas solution with mitochondrial ATP-sensitive potassium channel opener diazoxide on guinea pig papillary muscles protection after myocardial hypoxia. Methods: Twenty-four guinea pigs were randomly divided into three groups. In control group, cardioplegia was routine St.Thomas solution. In treatment group, cardioplegia was used modified St.Thomas solution. In blocker group, the muscle was treated with the specific potassium channel blocker glibenchamide 15 minutes before arrest used diazoxide cardioplegia. Myocardial electrophysical before and after cardioplegic arrest in guinea pig papillary muscles were studied. Results: 1, Time of recovery was shortened significantly in treatment group (P

[Summary] As the gradual deepening understanding of gastroesophageal reflux disease ( GERD ) , more extraesophageal symptoms are noted .The concept and therapeutic strategy for GERD has also undergone a quiet revolution over recent years .The application of proton pump inhibitor ( PPI) has been a landmark of medical treatment for GERD , and the invention of laparoscopic fundoplication and endoscopic radiofrequency energy delivery to the lower esophageal sphincter are marking a new era of comprehensive therapy for GERD .Good short and long term outcomes have been obtained since endoscopic radiofrequency energy delivery to the lower esophageal sphincter was applied on typical symptoms of GERD .Furthermore, the technique has also been successfully used in treating extraesophageal symptoms induced by proximal reflux in China .With more simple and less invasive features compared with anti-reflux surgery, the radiofrequency procedure has broad application prospects .

Objective To compare the impact of conventional vs. zero-balanced ultrafiltration on serum pro-inflammatory factors,acute kidney injury and clinical prognosis after cardio-pulmonary bypass procedure. Methods Forty patients receiving cardiac surgery under cardio-pulmonary bypass procedures in Xiangyang Central Hospital during January 2013 to June 2013 were randomly divided into conventional ultrafiltration group(group A,n=20)and zero-balanced ultrafiltration group(group B,n=20). Blood and urine samples were collected on different time points( pre-operation,T0;pre-ultrafiltration,T1;immediately after ultrafiltration,T2;24 hours post-operation,T3;48 hours post-operation,T4;7 days post-operation, T5). TNF-α,IL-6,KIM-1,CysC,serum creatinine and urea nitrogen were detected and compared. Pre-and post-operative clinical data were also collected. Results There was no difference in baseline data or intra-operative data(p>0. 05). TNF-αand IL-6 started to increase when the operation began. Compared with conventional ultrafiltration,zero-balanced ultrafiltration alleviated the increase of TNF-α and IL-6,espe-cially on T2,T3,and T4(p<0. 05). Zero-balanced ultrafiltration also decreased CysC,Rbp,serum creati-nine and urea nitrogen on T2,T3,and T4 compared with conventional ultrafiltration(p<0. 05). A total of 11 patients experienced AKI( >grade I)in group A and 2 patients experienced in group B(p<0. 01). There were significant differences of ventilation time,total complication incidence and ICU stay time be-tween two groups. There was no difference in other complications,post-operative days in hospital or death rate within 30 days. Conclusion Though there is a trend of more patients receving renal replacement therapy,no statistical difference has been achieved. In conclusion,zero-balanced ultrafiltration can effec-tively decrease the concentration of serum pro-inflammatory factors,alleviate acute kidney injury and improve the clinical prognosis after cardio-pulmonary bypass procedures. It is a safe and reliable method valuable for promotion.