Objective To investigate the optimal operative time interval from the combined application of endoscopic sphincteropapillotomy ( EST) and endoscopic nasobiliary drainage ( ENBD) to laparoscopic cholecystecto-my ( LC) in the treatment of concurrent cholecystolithiasis and choledocholith.Methods Retrospective analysis was made in the 180 patients who had received sequential treatment of concurrent cholecystolithiasis and choledocholith with the combined application of EFS+ENBD+LC.Patients in group A received LC 3 days after EST +ENBD;patients in group B received LC 7 days after EST +ENBD;patients in group C received LC 30 days after EST +ENBD.Comparative analysis was made in the duration of LC,the intraoperative blood loss,the number of conversion to laparotomy and the total hospital expenses among patients in the three groups.Results Operation time in group A,B and C was separately (34.70 ±2.24)min,(51.00 ±2.31)min and (34.43 ±2.51)min.Patients in group B suffered significantly-extended surgical duration ( F=6.15,P<0.01);Intraoperative blood loss in the three groups was separately (28.86 ±3.35)mL,(50.43 ±3.90)mL,(28.57 ±3.40)mL.Significant increase was observed in group B(F=7.23,P<0.01).There were 1,6 and 1 patients who underwent conversion to laparotomy in the three groups. Slight increase was found in group B(χ2 =6.54,P<0.05).Total hospitalization expenses was separately (10 093.29 ± 496.05)yuan,(10 845.86 ±744.27)yuan and (15 250.57 ±1 006.67)yuan.Patients in group C had incurred a significant increase(F=6.62,P<0.01).Statistical difference was equally found.Conclusion 3 days after EST+ENBD is the optimal operative time for LC.

Objective To explore the effects of α-keto acid combined with low protein diet on oxidative stress and electrolyte levels in patients with diabetic nephropathy. Methods Ninety-eight cases with type 2 diabetic nephropathy were randomLy divided into the treatment group(low-protein diet with α-keto acid 1 tablet,n=58) and the control group(normal protein intake,n=40). The 24 h urinary albumin excretion rate,glomerular filtra-tion rate ,serum oxidative stress (SOD ,MDA ,AOPPs) and electrolyte levels were measured before and after treatment. Results After 24 weeks of treatment,the 24 h urinary albumin excretion rate of DN patients in the treatment group was significantly lower than that before treatment and the control group (P < 0.05). However, there was no significant change in the glomerular filtration rate of the patients. Compared with the control group , the serum SOD activity of DN patients was significantly increased and the levels of MDA and AOPPs were decreased(P < 0.05)after 24 weeks of treatment. Simultaneously,the levels of serum potassium and phosphorus in the treatment group was decreased ,while the levels of calcium and magnesium were improved (P < 0.05). Conclusion α-keto acid combined with low protein diet can significantly reduce the urinary albumin excretion rate,improve the body′s antioxidant activity,and maintain serum electrolytes balance.

BACKGROUND: The prompt and efficient peripheral blood stem cell (PBSC) mobilization is necessary for repairing and regenerating injured tissues. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) or its combination with a large dose of chemotherapy is usually utilized for PBSC mobilization, but the regimen requires a long time and a complex procedure. OBJECTIVE: To modify the common PBSC mobilization, and investigate mobilization effect of PBSC in KM mice with short-duration and high-dose rhG-CSF. DESIGN, TIME AND SETTING: A randomized controlled animal trial was carried out in the laboratory of Hematology, the Second Affiliated Hospital to Dalian Medical University from September 2006 to March 2007. MATERIALS: Twenty-four KM male mice of 8 months old were divided into 3 groups according to the administration: conventional regimen group, short-duration and high-dose group, and saline control group. METHODS: Mice in the conventional regimen group were injected subcutaneously with rhG-CSF (0.2 ?g one time, twice a day for 6 days). Mice in the short-duration and high-dose group were injected with equal volume of normal saline at first 4 days, and then with rhG-CSF (2.5 ?g one time, twice a day for 2 days). Mice in the saline control group were injected subcutaneously with equal volume of normal saline for 6 days. MAIN OUTCOME MEASURES: Peripheral blood white blood cell (WBC) and colony-forming unit-granulocyte macrophage in KM mice were counted. RESULTS: WBC in the peripheral blood of KM mice increased rapidly in conventional regimen group and short-duration and high-dose group. No obvious change of WBC was observed in the control group. The number of colony-forming unit-granulocyte macrophage in conventional regimen group and short-duration and high-dose group was significantly elevated over 50 folds than control group (P 0.05). CONCLUSION: There is a good efficacy and feasibility in PBSC mobilization of KM mice with short-duration and large-dose rhG-CSF, and it is a successful amelioration of PBSC mobilization of mice peripheral blood.

Objective:To study the application value of different operation methods in the treatment of left hepatolithiasis in special position (the stone is located in the sagittal section of left portal vein and/or near the proximal end of left hepatic vein).Methods:From July 2015 to June 2018, 58 patients with left hepatolithiasis in special position in Department of General Surgery, the First People′s Hospital of Tianmen City of Hubei Province were analyzed retrospectively. According to the different operation methods, they were divided into two groups: left liver excision under laparoscopy and gallbladder incision stone group (left liver excision group, n=28) and laparoscopic left-half liver excision and bile tube excision stone removal group (left half liver excision group, n=30). The operation time, intraoperative bleeding, postopera-tive hospital stay, postoperative biliary fistula, postoperative bleeding, postoperative stone residue and stone recurrence were compared between the two groups.The measurement data with normal distributions were represented as ( Mean± SD), and comparison between groups was done using the t test. The chi-square test and Fisher precision test were used for comparison between counting data. Results:All 58 patients underwent laparoscopic surgery. Left liver excision group surgery time, in-serum bleeding, number of days hospitalized after surgery, number of postoperative bile fistula cases, number of postoperative bleeding cases were (161.53±30.56) min, (203.45±26.69) mL, (9.26±3.3. 86) days, 1 case, 1 case, left half liver excision group (153.16±42.63) min, (198.79±30.82) mL, (9.59±4.01) days 1 case, 1 case, The difference between the two groups was not statistically significant ( P>0.05). The number of postoperative stone residues was more than that of the left liver and outer leaf excision group (4 cases) than that of the left half of the liver excision group (1 case), and the number of postoperative stone recurrence cases was more than that of the latter (1 case), all of which were statistically significant ( P<0.05). The number of stone residues and stone recurrences after left liver excision group was 4 and 5 cases respectively, and the left half liver excision nurses were 1 case and 1 case, respectively, and the difference between the two was statistically significant ( P<0.05). Conclusion:For the left hepatolithiasis in special position (the stone is located in the sagittal section of the left branch of the portal vein and (or near the proximal end of the left hepatic vein in imaging), the lower abdominal left hemihepatectomy + choledocholithotomy is a safer and more effective treatment.

Objective　To prevent the after cataract induced by lens epithelial cells proliferation from postoperative cataract, monoclonal antibody (McAb) specific for rabbit lens epithelial cell is made, it will be the carrier further to be conjugated with cytotoxin. The conjugations will inhibit lens epithelial cells growth and not damage the other tissues of eye. Thereby McAb is the experimental bases of preventing after cataract.Methods　BALB/c mice were immunized by mixture of rabbit lens epithelial cells and Freund's adjuant. The immunized mouse spleen cells were fused with parental mouse myeloma cells (BALB/c　SP2/0) using polyethylene glycol (PEG-4000). The fused cells were selectively cultured by hypoxanthine, aminopterin and thymidine (HAT) culture medium. The specificities of the supernatant from hybridomas were tested by indirect immunofluorescence and immunohistochemistry SP (Streptavidin peroxdase conjugated method). The positive hybridomas were further cloned three times by methylcellulose culture medium to ensure monoclonality. At last, the consensual reaction of McAb was tested on human eye tissues.Results　Hybridomas were produced by fusion of spleen cells of immunized mice and mouse myoloma cells (SP2/0) with PEG-4000, and grown selectively in medium containing HAT after 16 days. Antibodies of the supernatant from hybridomas were tested on frozen sections of rabbit lens epithelial cell by indirect immunofluorescence. Only a positive clone secreted McAb against antigen of rabbit lens epithelial cell. The specificity of McAb was tested on paraffin sections of whole rabbit eye and whole human eye by immunohistochemistry SP. The results indicated that McAb was only positive to rabbit lens epithelial cell membrane and it was negative to the other tissues of rabbit or whole human eye tissues.Conclusions　McAb specific for rabbit lens epithelial cell was manufactured successfully. The specificity of McAb was strong. There was no consensual reaction on human eye tissues. It might be the experimental bases of further targeting chemotherapy on after cataract.

Objective To compare the influence for intravenous dexmedetomidine and midazolam during combined spinal and epidural anesthesia (CSEA) on sedation,respiratory and circulatory.Methods Ninety patients with lower extremity fractures and internal fixation,were divided into dexmedetomidine group,midazolam group and control group by random digits table with 30 cases each.CSEA was performed at L3-4 interspace.After block reached T8 level,dexmedetomidine,midazolam and 0.9％ sodium chloride were given to the three groups.Ramsay score,mean arterial pressure (MAP),heart rate (HR),partial pressure of carbon dioxide in end expiratory gas (PErCO2),respiratory rate (RR) were recorded before anesthesia(T0),after CSEA (T1),and 10,15,30,45,60 min after giving drug (T2-T6),and intraoperative awareness was recorded.Results Ramsay score in dexmedetomidine group and midazolam group at T2-T6 were higher than those in the group T0,T1 and concurrent control group (P ＜ 0.05),MAP were lower than those in the group T0,T1 and concurrent control group (P ＜ 0.05).HR in dexmedetomidine group at T2-T6 were lower than those in concurrent midazolam group and control group (P ＜ 0.05).PETCO2 in midazolam group at T2-T6 were higher than those in concurrent dexmedetomidine group and control group (P ＜ 0.05),RR were lower than those in concurrent dexmedetomidine group and control group (P ＜ 0.05).The rate of intraoperative awareness in dexmedetomidine group and midazolam group was lower than that in control group [16.7％(5/30) and 13.3％(4/30) vs.93.3％(28/30),P＜0.05].Conclusions Dexmedetomidine and midazolam provide good sedation to reduce intraoperative awareness,slight inhibition of blood pressure.Dexmedetomidine can decrease HR,but it does not influence respiratory function.Midazolam restrains respiratory function.

Objective To summarize the experience of application of artificial chordae transplantation in mitral valve repair.Methods One hundred and five patients with mitral regurgitation due to mitral degeneration,underwent mitral valve repair with artificial chordae transplantation.Operative technique included simple Gore-Tex artificial chordae transplantation in 25 cases,artificial chordae transplantation plus quadrangular resection of the posterior leaflet in 67 cases,artificial chordae transplantation plus quadrangular resection of the posterior leaflet and Sliding technique in 13 cases.Results No early death occurred after operation.The postoperative complications were caused in 30 patients including sinus bradycardia in 8 cases,supraventricular tachycardia in 20 cases,late cardiac tamponade in onecase,pacemarker implantation in one case.Hospital time was 9-21 (14 ±4) d.Follow-up was done to 96 patients for 3 months to 5 years with a follow-up rate of 91.4％(96/105).Among 96 cases,one died of cerebral infarction after 13 months,one died from accident while the remaining were alive,74 cases were with cardiac function (NYHA) of grade Ⅰ and 20 cases with grade Ⅱ.No regurgitation was found in 92 cases,mild regurgitation in 2 cases.There were no artificial chordae ruptures.Conclusions The key to improve the early and midterm results of artificial chordae transplantation are to choose patients strictly,to grasp proper surgical skills,to do a good job in intraoperative shaping effect evaluation and the myocardial protection during extracorporeal circulation.

BACKGROUND:Artificial chordae transplantation and saddle ring annuloplasty are the key steps in mitral valvuloplasty. However, there are no large-size studies addressing the safety, validity and efficacy of these treatments in China.
OBJECTIVE:To summarize the efficacy and safety of artificial chordae transplantation and saddle ring annuloplasty in the treatment of degenerative mitral regurgitation.
METHODS:Eighty-five patients with degenerative mitral regurgitation underwent mitral valve repair with artificial chordae (Gore-Tex sutures) transplantation and saddle ring (SJMTM rigid saddle-shaped) annuloplasty from January 2009 to May 2013 in General Hospital of Shenyang Area Military Command, China. Operative technique included simple artificial chordae transplantation in 41 cases, artificial chordae transplantation plus quadrangular resection of the posterior leaflet in 22 cases, artificial chordae transplantation plus quadrangular resection of the posterior leaflet and sliding technique in 23 cases. Al cases received saddle ring annuloplasty.
RESULTS AND CONCLUSION:Fol ow-up after treatment was done among 78 patients for 6 months to 4 years, with a fol ow-up rate of 91.7%(78/85). Among the 78 cases, one case died of cerebral infarction after 13 months, one died from accident, and the remaining 76 were alive. According to the evaluation of cardiac function (NYHA), 59 cases were in grade I and 17 cases were in grade II. Color ultrasound displayed that, no regurgitation was found in 67 cases, and mild regurgitation in 9 cases. The echocardiography showed that postoperative left atrium diameter (P<0.05 or P<0.01), left ventricular end-diastolic diameter (P<0.05 or P<0.01), left ventricular end-systolic diameter (P<0.05 or P<0.01), and the ratio of regurgitation beam area and left atrial area (P<0.05 or P<0.01), mean pulmonary artery pressure (P<0.05 or P<0.01) were significantly decreased compared with that before operation. Ejection fraction was significantly increased after operation (P<0.05 or P<0.01). No systolic anterior motion occurred. The postoperative complications included sinus bradycardia in 12 cases and paroxysmal supraventricular tachycardia in 25 cases, late cardiac tamponade in 1 case at 1 week postoperatively, and pacemarker implantation in 1 case (who exhibited bradycardia-tachycardia syndrome before operation). There was no ring rupture, ring avulsion, hemolysis, left ventricular outflow tract infarction and artificial chordae rupture or splitting. No cases needed reoperation on valve replacement. Application of artificial chordae transplantation and saddle ring annuloplasty is a safe and effective means for treating degenerative mitral regurgitation, with excellent midterm outcomes.

Objective:To investiate biological properties of Mesenchymal stem cell(BMSC) from rat bone marrow in vitro,and their effects in inducing immune tolerance.Methods:BMSCs were obtained from rat bone marrow by density gradient centrifugation and selected by cell attachment.The morphology of BMSCs was observed by electron microscope.The expression of surface molecules of CD34 and CD44 was analyzed by flow cytometry.The cells were injected ex vivo intravenously after in vitro culture and then CD4+ CD25+ Treg ratio were determined.The proliferation of thymic cells was estimated by 3H-TdR incorporation method.Results:BMSCs were fibroblast-like cells.The expression ratio of CD44 in BMSCs was 98%,but expression of CD34 was negative.After administration of BMSCs,the ratio of CD4+CD25+Treg(2.5%?0.69%) from peripheral blood of rats was higher than in normal group(0.8%?0.14%) and PBS control group(1.0%?0.23%),P

Objecive To explore the significance of RECK expression in breast cancer .Methods Im-munohistochemical staining was used to analyze RECK expression levels in patients with breast cancer .We com-pared these data with the clinicopathological features of these patients .Rseults Breast cancer patients with nega-tive RECK expression had significantly lower DFS and 5-year survival rates than patients with positive RECK expression.In addition,for node-negative breast cancer ,negative RECK expression indicated markedly unfavor -able survival rate than positive arm .Multivariate analysis further confirmed that RECK expression was an inde -pendent prognostic factor for patients with breast cancer .Conclusion The loss of RECK expression indicates un-favorable survival rate for patients with breast cancer .RECK expression is a new ,important risk factor for recur-rence in breast cancer .