Objective To evaluated the preserving or removing jejunal mucosa pancreatico-jejunal invaginated anastomosis in pancreaticoduodenectomy.Methods Between February 2003 and December 2012,58 patients underwent pancreaticoduodenectomy using pancreaticojejunal invaginated anastomosis in our department.In group A,28 patients received pancreatico-jejunal invaginated anastomosis using jejunum preserving technique.Briefly,3 to 4 cm remanent pancreas was inserted into jejunum and thc jcjunum was bundled up using No.7 silk thread at 2 to 3 cm distant to the cutting surface of the pancreas.In group B,30 patients received pancreatico-jejunal invaginated anastomosis using jejunum removing technique.Briefly,the jejunum mucosa was everted and the proximal 3 cmlong mucosa was removed.Then the everted jejunum was re-positioned and the ending of the jejunum was interruptedly sutured to the remanent pancreas.Finally,the covered jejunum was tied up as the group A.Results The pancrcatico-jejunal anastomosis time was 36 ±0.34 minutes shorter in group A than group B (P ＜0.001).The incidence of pancreatic fistula was higher in group B (20.0％) than group A (0％).In contrast,the incidence of post-operative pancreatic bleeding was compared between the two groups (3.6％ vs 3.3％,P =1.000).Conclusions Preserving ejunum mucosa pancreatico-jejunal invaginated anastomosis was unaffected by pancreatic texture pancreatic duct size and position.This method takes the advantages of simple operation and reducing the incidence of pancreatic fistula.

Objective:To observe the changes of choriocapillary blood flow area (CBFA) and diameter of ellipsoid zone absence (DEZA) of eyes with idiopathic macular hole (IMH) before and after surgery and its correlation with visual function.Methods:A prospective study. From May 2019 to January 2020, 23 IMH patients (IMH group and fellow eyes group) diagnosed in the Department of Opthalmology of The First Affiliated Hospital of Bengbu Medical College, and 30 age and gender-matched normal controls (30 eyes, control group) were included in this study. All patients were examined the best corrected visual acuity (BCVA) before and after surgery. The BCVA examination was performed using the international standard Snellen visual acuity chart, which was converted into logarithm of the minimum angle of resolution (logMAR) BCVA for record. CBFA and DEZA were obtained by optical coherence tomography angiography (OCTA). CBFA was measured on the choriocapillary with a circle of 1.0 mm radius centered in the fovea. DEZA was the diameter of ellipsoid zone absence horizontally. The differences of CBFA and logMAR BCVA in the three groups, and the changes of CBFA, DEZA, logMAR BCVA in IMH group before and after surgery were analyzed. The measurement data between the three groups were compared by one-way analysis of variance, and the independent samples t test was used for the comparison between the two groups. Pearson correlation analysis was used for the correlation between the index, multiple linear regression analysis were performed between postoperative visual acuity and preoperative parameters. Results:Before and 6 months after surgery, the logMAR BCVA in IMH group were 1.26±0.7 and 0.48±0.22, CBFA were 1.49±0.30 and 1.92±0.17 mm 2, DEZA were 1 080.22±576.98 and 433.78±423.04 μm, respectively. Compared with those before surgery, BCVA and CBFA were significantly improved, DEZA obviously decreased and the differences were statistically significant ( t=5.53, 7.77, 6.58; P<0.01). CBFA in IMH group was lower than fellow eyes group and control group, the difference was statistically significant ( F=14.13, P<0.01). After surgery, CBFA in IMH group increased similar to that of fellow eyes group ( t=4.32, P=0.37), but lower than that of control group with the significant difference ( t=4.07, P<0.01). The results of Pearson correlation analysis showed that logMAR BCVA and DEZA were positively correlated ( r=0.69, 0.75; P<0.01), CBFA and DEZA were negatively correlated ( r= -0.49, -0.89; P<0.05) before and after surgery. Multivariate linear regression analysis showed that the postoperative logMAR BCVA was positively correlated with DEZA ( t=2.32, P=0.02). Conclusions:After surgery, BCVA and CBFA of IMH eyes are significantly increased, DEZA is obviously reduced. Before and after surgery, logMAR BCVA is positively correlated with DEZA, and CBFA is negatively correlated with DEZA.

Objective To investigate whether tissue engineered bone with implanted vascular bun-dles can up-regulate release of vascular endothelial growth factor (VEGF) in models of femoral defect in rabbits.Methods Thirty-two rabbits were randomized into 2 even groups.In both groups, a segmental bone defect of 15 mm in length was made at the left femur before a tissue engineered bone was inserted into the defect.In the experimental group, a femoral vascular bundle was implanted into the tissue engineered bone.In the control group, there was no vascular implantation.At 2, 4, 8, and 12 weeks after implantation, samples were taken to determine new bone formation by histology and expression level of VEGF by immuno-histochemistry.Results The new bone formation was significantly higher in the experimental group at the end of 4, 8, and 12 weeks(P < 0.05) .The expression level of VEGF in the experimental group was also significantly higher than in the control group at all time points after operation, and the expression of VEGF peaked at 4 weeks.Conclusion Tissue engineered bone with vascular bundle implanted can up-regulate VEGF release in models of femoral defect in rabbits.

BACKGROUND: Neuropeptides, a kind of endogenous active substance in nerve tissues, can modulate physiological functions of multiple body systems.OBJECTIVE: To observe the effects of vascular bundles or sensory nerve tracts implanted into tissue-engineered bone for rabbit large bone defects on the expression levels of calcitonin gene-related peptide (CGRP) and neuropeptide-Y.METHODS: Fifty-four New Zealand rabbits were enrolled to make model of large bone defects, and then, the animal models were randomly divided into three groups, including sensory nerve tract, vascular bundle, and control groups (n=18 per group), followed by implanted with sensory nerve tracts, vascular bundle, and tissue-engineered bone without sensory tracts or vascular bundle, respectively. The defected bone received gross and Masson staining at 4, 8 and 12 weeks after modeling, to compare the expression levels of CGRP and neuropeptide-Y in each group.RESULTS AND CONCLUSION: mRNA expression levels of CGRP and neuropeptide-Y in the sensory nerve tract and vascular bundle groups were significantly higher than those in the control group at different time points after modeling (P < 0.05). mRNA expression levels of CGRP and neuropeptide-Y in the tissue-engineered bone began to be increased and peaked at the 8th week, and then decreased (P < 0.05), which were the lowest at the 4th week (P < 0.05).Immunohistochemical staining results showed that CGRP was mainly found in the bridge, periosteum of newly born bones and around blood vessels; while neuropeptide-Y mainly localized in the medullary cavity and around blood vessels. These results indicate that the implantation of vascular bundle and sensory nerve tracts for bone defects can upregulate the expression levels of CGRP and neuropeptide-Y, and promote bone repair. However, sensory tract implantation may cause sensory impairment; thereafter, vascular bundle implantation is more suitable for ideal tissue-engineered construction to meet physical requirements.

Objective To explore the surgical methods for the treatment of the total anomalous pulmonary venous connection (TAPVC) and choices to prevent related complications.Methods We analyzed retrospectively the clinical data of 24 cases with TAPVC admitted to our hospital from Jan 2006 to Dec 2011,including 15 male and 9 females with the age range of 50 d-14 years.There were 10 cases younger than six months,accounting for 41.7％ (10/24).The average body weight was (9.30 ± 3.96) kg.There were 8 cases 33.3％ (8/24) had a body weight of below 10.00 kg.Among the patients,16 cases (66.7％,16/24) were supracardic type,6 (25.0％) were cardiac type,and 2 (8.30％) were intracardiac type.For the treatment of the upracardiac type,5 cases were treated through the right atrium and interatrial septum incision path;Eleven cases were through the left atrial anastomosis.For the treatment of the cardiac type,the right atrial incision was used for coronary vein antrum isolation,and the patch was carefully packaged to separate the coronary sinus openings into the left atrial side.For the 2 cases of the intracardiac type,heart was slightly lift towards the right,and the venous anastomosis was performed for the left atrial posterior wall and the summary vein,and the vertical vein was then ligated.Results No surgery-related death occurred.Reoperation was performed for 1 patient occurred pulmonary edema due to pulmonary venous obstruction induced two days post-surgery.Condition was improved after the extension of left atrial side as the patient was found to have anastomotic stenosis.Postoperative arrhythmia were observed in 7 cases (29.2％,7/24),including 3 nodal arrhythmia (12.5％,3/24) and 4 (16.7％,4/24) atrial arrhythmia.Patients were followed up for 4-24 months.All children during the follow-up period were in good condition.They had significantly improved activity tolerance compared with pre-surgery.Chest X-ray showed clear markings free of congestion.Conclusion Appropriate surgical approach and routes could help improve the success rate of surgery treatment of TAPVC and reduce postoperative complications,thus achieving good therapeutic effect.

Objective The relationship between hypertension ( HTN) and ischemic stroke recurrence is unclear , but there may be different effects of HTN on the risk of recurrence .This study aims to explore whether HTN contributes differently to the recur-rence among subtypes of ischemic stroke ( IS) . Methods We eventually enrolled 1114 patients with ischemic stroke from Jul 2008 to Dec 2012 registered in Nanjing Stroke Registry Program (NSRP) in this study.All the patients were classified according to Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria: 315 (28.3%) patiwnts were classified as Large-artery atheroselerosis (LAA), 212 (19.0%) as cardioembolism (CE), 266 (23.9%) as small-artery occlusion (SAO), and 321 (28.8%) as other de-termined and undetermined etiologies ( Other) .The association between HTN and stroke recurrence in patients with different IS sub-types was analyzed using multivariate Cox regression analysis . Results The average follow-up duration was (19.4 ±10.3) months. Of 1114 patients with IS, 158 (14.2%) patients experienced a recurrent stroke .Patients with HTN had a significantly higher stroke recurrence rate than those without (16.5%vs 10.5%, P<0.05).Multivariate Cox regression analysis indicated that HTN increased the risk of ischemic stroke recurrence (HR=1.722, 95%CI:1.181-2.512, P=0.005).After stratification by TOAST subtypes, analysis revealed an association between HTN and stroke recurrence in LAA( HR=3 .767, 95%CI:1.866-7.585, P=0.001) and SAO (HR=3.530, 95%CI:1.156-12.740, P=0.028), but not in the other subtypes (CE: HR=0.773, 95%CI:0.370-1.615, P=0.493;Other:HR=1.498, 95%CI:0.590-3.807, P=0.395). Conclusion HTN is an independent risk factor for recurrent ischemic stroke and is related to the recurrent ischemic stroke in patients with large-artery and small-vessel disease .

Objective To investigate the inhibitory effects of L. monocytogenes expressing melanomainhibiting activity(MIA)gene on malignant melanoma.Methods The plasmid pERL3-hly-MIA was constructed and used to transform live L.monocytogenes by electroporation.Atier culture.the transformed L.monocytogenes,namely LM-hly-MIA,was harvested for the detection of MIA protein by Westem Blot.A total of 24 mice were divided into three groups,namely control group,LM group and LM-hly-MIA group,to be immunized with physiological saline,L.monocytogenes and LM-hly-MIA,respectively.One day after the immunization.mice were subcutaneously inoculated with 0.1 mL of B16 cells at a concentration of 1×107/mL.One week later.reimmunization was performed.The size and weight of tumors were observed and measured in these mice.Results Enzyme digestion and Western blot confirmed the Successful construction of plasmid pERL3-hly-MIA.The average weight of tumors in mice was 4.33±0.91 gram.3.36 ±0.41 gram and 1.89±0.52 gram,respectively in the control group,LM group and LM-hly-MIA group,respectively(P＜0.05).with the inhibition rates of tumor growth being 22.4%and 61.5%in LM group and LM-hly-MIA group.respectively.Conclusion The attenuated strain of L.monocytogenes expressing MIA gene could evidently inhibit melanoma growth.

Objective To explore whether the respective implantation of vascular bundles and sensory nerve tracts into a tissue-engineered bone will affect the expression of CGRP (Calcitonin gene related peptide) and its receptor. Methods Fifty-four New Zealand rabbits were randomly divided into 3 even groups for implantation of sensory nerve tracts (group A),implantation of vascular bundles (group B),and a control group of simple tissue-engineered bone (group C) . Animals were sacrificed 4,8,12 weeks after implantation,respectively. Masson staining was conducted to observe the process of bone formation and re-molding. CGRP and CGRPR-1 expressions in the new bone were measured by immunohistochemistry and Real-time PCR at 4,8 and 12 weeks after implantation. Results At all time points,the CGRP and CGRPR-1 expressions in groups A and B were significantly higher than in group C (P<0.05),and those in group A were higher than in group B too (P<0.05) . Over time,the expressions of CGRP and CGRPR-1 mRNA in each group in the new bone tissue were gradually reduced after an initial increase. The neuropeptide expression at the 8th week was higher than those at the 4th and 12th weeks. The neuropeptide expression at the 4th week was the lowest. The expression of CGRP was mainly localized in the periphery of newly generated bone,periosteum and the blood vessels. The expression of CGRPR-1 was mainly localized in the periphery of osteoblasts. Conclusions Implantation of either vascular bundles or sensory nerve tracts can promote neuropeptide secretion. The vascular bundle implantation may result in higher expressions of CGRP and CGRPR-1 than sensory nerve tract implantation.

Objective To investigate the inhibitory effect of live-attenuated Listeria monocytogenes (LM)-based vaccines expressing the gene encoding a melanoma differentiation antigen,MART-1,on malignant melanoma and their mechanism.Methods The constructed plasmid pERL3-MART-1 was used to transform live-attenuated LM by electroporation to construct recombinant LM.i.e.△inlB LM-MART-1 and △actA/△inlB LM-MART-1.The half lethal dose (LD50) of attenuated listeria strains was determined by concentration gradient dilution method.C57BL/6 mice were randomly divided into three groups,namely PBS group,△inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group.Mice were inoculated by intraperitoneal injection of O.1 LD50 of each rLM strain or PBS only.One week later,the mice were injected subcutaneously with 1×105 B16F10 cells(a mouse melanoma cell strain)in 200μl of PBS.Reimmunization was performed on day 14 and 21.Subsequently,the growth of tumor and survival of tumor bearing mice were observed.All mice were killed on day 28,and tumor tissue as well as splenocytes were obtained from these mice for the detection of MART-1 gene expression by real-time quantitative PCR and the percentage of CD4+CD25+T cell by flow cytometry.Results The recombinant △inlB LM-MART-1 and △actA/△inlB LM-MART-1 were constructed successfully.The LD50 of △inlB LM and △actA/△inlB LM was lower than LM-EGDe by 100 and 10 000 times respectively.Compared with PBS,the tumor growth was inhibited with △inlB LM-MART-1 by 46.95%(F=6.3,P<0.05),and by 83.96% with △actA/△inlB LM-MART-1(F=37.8,P<0.01).The relative expression level of MART-1 in △inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group was 8.988±0.207 and 11.315±0.445 times that in PBS group (both P<0.05).The percentage of CD4+CD25+T cells in splenocytes was (2.52±0.20)%,(1.14±0.13)% and (0.44±0.15)% in PBS group,△ialB LM-MART-1 group and △actA/△inlB LM-MART-1 group,respectively;the differences were statistically significant between the three groups (all P

Listeria monocytogenes (LM), a Gram-positive facultative intracellular bacterium, can be used as an effective exogenous antigen expression vector in tumor-target therapy. But for successful clinical application, it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen. In this study, attenuated LM and recombinants of LM expressing melanoma inhibitory activity (MIA) were constructed successfully. The median lethal dose (LD(50)) and invasion efficiency of attenuated LM strains were detected. The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma. The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction (PCR) with specific sequence, meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot (ELISPOT) assay. The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM, and attenuated LM expressing MIA, especially the double-genes attenuated LM recombinant, could significantly induce anti-tumor immune response and inhibit tumor growth. This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.