The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.

Objective To discuss the advance and experience of postoperative management in multivisceral transplantation(MVT).Methods Five patients with abdominal multiple organs malig- nances were subjected to multivisceral transplantation.The excised specimens included the liver,pan- creas,duodenum,stomach,spleen,mesentery and all of the involved lymph nodes.The transplanted organs were liver,pancreas,duodenum and parts of intestine.Patients were monitored and sustained stably in breath,hemodynamics and homoeostasis.The coagulation was also monitored and kept in balance.The intensive immunosuppressive protocol,including steroid,tacrolimus,mycophenolate mofetil and Zenapax,was used to prevent the acute rejection.The endocrine and exocrine of pancreas were tested,color Doppler examination was done every day,and Somatostatin was used to prevent pancreatitis.Drugs to bacteria,fungus and virus infection were prophylactically used.Results All patients waked up with 4-6 h after operation and began recovering with 3-7 days.Dynamic examina- tions revealed that the functions of grafted liver,pancreas and duodenum were satisfactory.No com- plications of acute rejection,pancreatitis,bleeding and thrombosis occurred.Conclusion Many ques- tions in postoperative management related to multivisceral transplantation are devoted to study.Nice experience of management to organ transplantation,detailed monitoring and prophylactically medica- tions are safeguards to steady recovering and further prognosis.

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Objective To explore the clinical characteristics and management of acute myocardial infarction (AMI) early after kidney transplantation (＜3 months).Method Five cases of AMI early posttransplantation among 122 kidney transplant recipients from June 2011 to December 2012 were retrospectively reviewed.Results Of 5 AMI patients,there were 2 cases within one week postoperatively,one case at 11 th day postoperation,and the other two at 29th day and 46th day after operation respectively.Acute left heart failure was complicated in 3 cases within first two weeks.All the AMI patients had elevated TnⅠ levels which declined subsequently.The climax of TnⅠ levels in all the 5 AMI patients were above 5 ng/mL,and more than 20 ng/mL in two AMI patients within one week.Given by symptomatic and supportive treatment,antiplatelet and anticoagulation therapies and cardioprotective medications,all the five AMI patients were improved.Low molecular heparin was additionally administrated to the 2 cases within first week according to the severe conditions.New emerged small volume of hematocele was proved by ultrasound after 3 days and low molecular heparin was ceased.All the 5 patients survived and neither thrombolysis nor percutaneous coronary intervention therapy was given to them.Conclusion In addition to general prevention against AMI in kidney recipients with high risk factors,managing anemia and hypertensiorn,and improving graft function and systematic status are also important to decrease the risk of AMI.Moreover,cardioprotective therapy including antiplatelet therapies,beta-blockers,angiotensin-converting enzyme inhibitors (ACEI)/angiotensin-2 receptor blockers and statins,which are recommended to the general population with AMI,will also profit to the kidney transplant recipients with AMI.However,aggressive intervention therapies might be more prudent to be used in this population.

This paper expounds the concept and connotation of the nurse professionalism, introduces the re-search status of foreign nurses professionalism assessment tools,discusses the influence factors of nurse professional-ism,including:personal factors, organizational factors and occupational factors.It also puts forward following sugges-tions:constructing the nurse professionalism of theoretical system and evaluation methods, advocating the spirit of nurses professionalism cultivation,creating a health care professional environment,providing the reference the devel-opment of our country's theory and improve the nurse professionalism.

Environmental Pollution ; prevention & control ; Health Education ; Health Knowledge, Attitudes, Practice ; Humans ; Iatrogenic Disease ; prevention & control ; Medical Laboratory Personnel ; psychology

Objective To investigate the clinical value of a new anvil inserting method for esophagogastrostomy or esophagojejunostomy during laparoscopic radical proximal gastrectomy or radical total gastrectomy for gastric cancer.Methods The clinical data of 21 patients with gastric cancer who received laparoscopic radical proximal gastrectomy or radical total gastrectomy at the Southwest Hospital from March 2010 to February 2011 were retrospectively analyzed.Five trocars were inserted through the abdominal wall of the patients.After perigastric lymphadenectomy and mobilization of esophagus,an incision was made on the esophagus above the tumor,and then the anvil with drawn wire attached was inserted into the esophagus.An endo-cutter was applied to cut the esophagus adjacent to the incision left the drawn wire untouched,and then the stem of the anvil was pulled out by the drawn wire for laparoscopic anastomosis. Results The operations were successfully accomplished under the laparoscope with no conversion to open surgery.Fifteen patients received laparoscopic radical total gastrectomy and 6 received laparoscopic radical proximal gastrectomy. The mean operation time,volume of blood loss,time to off-bed activity,passage of flatus and postoperative duration of hospital stay were (257 ± 38) minutes,( 119 ± 32) ml,(2.5 ± 0.5 ) days,( 3.7 ± 0.8 ) days and (7.5 ± 2.6) days,respectively.No perioperative mortality,anastomotic bleeding or anastomotic fistula was detected.One patient was complicated with pulmonary infection + pleural effusion and was cured by conservative treatment; 1 was complicated with anastomotic stenosis which was alleviated by gastroscopic balloon dilation; 1 was complicated by incisional infection and was cured by medical treatment after drainage.No cancer cells were detected at the anastomotic ring or resection margin of the specimen.There were 4 patients with well-differentiated adenoma,8 with moderate-differentiated adenoma and 9 with poor-differentiated mucinous adenoma.There were 5 patients in stage Ⅰ,10 in stage Ⅱ and 6 in stage Ⅲ (UICC staging).Twenty-one patients were followed up for a mean period of (11 ±4) months (range,6-17 months ),no tumor recurrence or metastasis was detected. Conclusions The new technique for anvil insertion is safe,effective and easy for manipulation and learn.It offers a new approach for laparoscopic digestive tract reconstruction.

OBJECTIVETo explore the effect of drug-containing serum of Chinese herbal compounds [Xiongshao Capsule (XS, for activating blood) and Huanglian Capsule (HL, for dispelling toxin)] on tumor necrosis factor-alpha (TNF-alpha)-induced adherence between human umbilical vein endothelial cells (HUVECs) and polymorphonuclear neutrophils (PMN), inflammatory reaction and expression of related proteins in mitogen-activated protein kinase (MAPK) pathway.

METHODSThirty-two rats were randomly divided into four groups (8 in each group) using random digit table: the blank control group treated with distilled water, the test group I treated with Chinese herbal compound of XS (0.135 g/kg), the test group II treated with Chinese herbal compound of HL (0.135 g/kg), and the test group Ill treated with Chinese herbal compound of XS (0.135 g/kg) and HL (0.135 g/kg). All medication was given by gastrogavage once a day for a week. Rats' blood serum was harvested 1 h after the last administration to prepare drug-containing serum. HUVECs were exposed to TNF-alpha (100 ng/mL) to induce cell injury model and incubated with corresponding drug-containing serum (10%) for 24 h. Normal rats' serum was given to cells in the blank control group and the model group, while XC + HL containing serum was given to cells in the rest 3 groups. The adherence of HUVECs and PMN cells was detected by using rose bengal strain. Levels of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and interleukin-1beta (IL-1P) in the supernatant of cultured HU-VECs were determined by ELISA. Protein expressions of mitogen-activated protein kinases p38 (p38MAPK) and extracellular signal-regulated kinase 1/2 (ERK 12) were determined by Western blot.

RESULTSCompared with the blank control group, HUVECs were seriously injured; PMN adherence amount significantly increased; levels of E-selectin, ICAM-1, and IL-1beta increased; expression levels of p-p38MAPK and p-ERK 1/2 in the supernatant of HUVECs significantly increased in the model group (all P < 0.01). Compared with the model group, HUVECs-PMN adherence amount decreased (P < 0.05); levels of E-selectin, ICAM-1, and IL-1 beta in the supernatant of HUVECs decreased (P < 0.01, P < 0.05); expression levels of p-p38MAPK and p-ERK 1/2 of endothelial cells decreased in the test group I, II, and III (P < 0.01).

CONCLUSIONSDrug-containing serums of activating blood, activating blood and dispelling toxin could attenuate TNF-alpha induced injury of HUVECs, inhibit HUVECs-PMN adherence and the release of adhesion factors. Its mechanism might be involved with protein phosphorylation of p38MAPK and ERK 1/2 in the MAPK pathway.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; E-Selectin ; Endothelial Cells ; physiology ; Human Umbilical Vein Endothelial Cells ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1beta ; Mitogen-Activated Protein Kinase 3 ; Neutrophils ; Rats ; Serum ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.

Objective Rapid elevation of the IgG antibody against Candida Enolase ( Eno ) has been observed in patients with invasive candidosis in an early stage .The present study was to confirm the association of Candida colonization with humoral im-mune anamnestic response of the dominant antigen . Methods Twenty-four mice were randomized into group 1 treated by oral Candi-da colonization plus intraperitoneal infection ( immunocompetent , n=8) , 2 treated with immunosuppressant in addition to the treatment of group 1 ( immunocompromised , n=8) , 3 treated by oral Candida colonization only ( immunocompetent , n=4) and 4 treated by in-traperitoneal injection only( immunocompetent, n=4).The number of Eno-specific memory B-cells in the spleen and the levels of IgG , IgM and IgA antibodies were determined in the peripheral blood of the immunocompetent and immunocompromised invasive candidiasis mice . Results At 7 days after invasive infection , there were significantly more Eno-specific memory B-cells in the mice of groups1 ( 47.25 ± 13.81) and 2 (43.14±15.95) than in groups 3 (8.00±3.74) and 4(8.50±2.38) (P<0.01), with no statistically significant differences between either groups 1 and 2 or groups 3 and 4 (P >0.05).Eno-IgG antibodies were detected in the serum of the mice of the first two groups in the early stage of invasive infection and positively corre -lated with antigen-specific memory B-cells (r=0.737,P <00.1 ). Conclusion Rapid elevation of the Eno-IgG antibody level in the early stage of invasive infection after Candida colonization may be attributed to the rapid proliferation of humoral immune memory cells.