Objective To evaluate a surgical approach and anastomosis for the treatment of carcinoma of the gastric cardia. Methods Transabdominal intramediastinal esophagogastric anastomosis covered by sero-muscular flap of gastricwall for cardial carcinoma in 187 cases. Results The method reached the satisfactory surgical result in terms of tumor free cut edge on esophagus end of the resected samples. And the morbidity rate was 5.8%. Conclusion The technique enables lymphadenectomy within the lower mediastinum and a sufficiently long enough resection of esophagus. Transabdominal incision of the crus dextrum of the diaphragm makes a clear operative field for the purpose of radical operation for carcinoma of the gastric cardia. The anastomosis effectively prevents anastomotic leakage. This procedure is indicated for cardial carcinoma cases in which the esophaged involvement is within 2cm.

Objective:To investigate the Migration ability toward human pancreatic carcinoma cell line and human colon carcinoma cell line with difference HSP 70 plasma membrane expression .Methods: CD3-CD56+NK cells were obtained from human peripheral blood mononuclear(PBMC)in stem cell growth medium SCGM,2μg/ml TKD was added to the medium on 10th day,the ac-tivating receptor CD94/NKG2C expression levels on NK cells was detected with FAC after 4 days.The human pancreatic carcinoma cell line Colo357 and the human colon carcinoma cell line CW 2 were separated into Colo+and CW2+with high HSP70 expression and Colo-and CW2-with low HSP70 expression;Migration assays of NK to the four difference cell lines were performed in a Transwell cell culture system.The cytolytic activity of TKD-activated NK cells against the four subline with HSP 70 expression on their cell surface was analyzed by MTT assay.Results:Flow cytometry analysis showed that CD 3-CD56+NK cells could expanded after 2 weeks in SCGM medium,and the largest percentage of NK cell was (92.50 ±1.25 )%.CD94 expression levels on NK cells increased obviously after TKD inducement the cell surface HSP 70 expression of Colo+, Colo-were ( 78.2 ±2.2 )% and ( 27.3 ±1.2 )% separately , the cell surface HSP70 expression of CW2+,CW2-were (91.1±2.5)%and (18.2±1.0)%separately after FACS;the Migration of NK cells toward Colo+was (68.6±2.8)%,higher than the migration toward Colo-with (22.8±1.5)%;the Migration of NK cells toward CW2+was(73.5±2.7)%,higher than the migration toward CW2-with (18.2±1.3)%;the cytolytic activity of NK against Colo +was(61.2± 3.0)%compared to (24.5 ±1.5)%against Colo-when the ratio of effector cells and target cell was 20 ∶1,the cytolytic activity of NK against CW2+was (63.8±3.2)%compared to (22.4±1.8)% against CW2-when the ratio of effector cells and target cell was 20∶1.Conclusion:TKD-activated NK cells are highly efficient cytolytic effector cells which have stronger significant migration toward HSP70-positive tumor target cells on their cell surface in vitro .

Objective:To investigate the antitumor effect of TKD-activated NK cells on tumor growth inhibition of human pancreatic carcinoma in nude mice .Methods:CD3-CD56+NK cells were obtained from human peripheral blood mononuclear ( PBMC) in stem cell growth medium SCGM , 2 μg/ml TKD was added to the medium on day 10.The activating receptor CD 94/NKG2C expression levels on NK cells was detected with FAC after 4 days.The cytolytic activity of TKD-activated NK cells against human pancreatic carcinoma subline with HSP 70 expression on their cell surface was analyzed by MTT assay .Established a new model of orthotopic-transplantation tumor of human pancreas .NK cells were injected i.v.into the tail vein of tumor-bearing mice on day 15,the antitumor activity of the NK were evaluated .The capacity to infiltrate Colo 357 tumors in SCID/beige mice was detected with Immunohis-tochemistry.Results:Flow cytometry analysis showed that CD3-CD56+NK cells could expanded in SCGM medium ,and the average percentage of NK cell was (87.50 ±1.35 )%.CD94 expression levels on NK cells increased obviously ,the mean fluorescence intensity of CD94 was(220.56±1.82),compared to (68.72±1.85)of control group cell.The cytolytic activity against HSP70 membrane-positive pancreatic carcinoma sublines Colo 357 cells was high and there was significantly statistical difference between TKD-activated NK cells and unactivated NK cells.The cytolytic activity was(68.72±2.55)%when ratio of effector cells and target cell was 40:1.TKD-activated NK cells had a stronger suppressive effect on tumor growth in BALB /c nude mice bearing Colo 357 cells in vivo ,Median inhibitory rates was ( 61.3 ±1.5 )% .There was significant statistical difference compare to control group ( P <0.01 ) .The result of Immunohistochemistry indicated that predominantly NK cells induced with TKD had the capacity to infiltrate Colo 357 tumors in SCID/beige mice.Conclusion: TKD-activated NK cells are highly efficient cytolytic effector cells which have a stronger significant suppression against pancreatic carcinoma growth in vivo .

Objective To investigate the clinical effects of the posterior atlantoaxial pedicle screw fixation combined with unilateral axial spinous process and lamina screws and allogeneic bone graft for atlantoaxial instability.Methods From March 2010 to April 2014,data of 10 patients with atlantoaxial instability who had undergone atlanto-axial vertebral pedicle fixation and interbody fusion combined with posterior atlanto-axial spines lamina nails were retrospectively analyzed.There were 6 males and 4 females with an average age of 39 (range,16-62) years old.The inclusion criteria was atlantoaxial instability together with unilateral vertebral artery segment high cross deformity (7 patients) or atlantoaxial joint break bad involving unilateral pedicle (7 patients).3 patients have spinal cord disease symptoms and physical signs,both suffered limited neck mobility and pain.The VAS scores were 1-8,with an average of 3.70±2.11.Preoperative X-ray,CT three-dimensional reconstruction and MRI were collected.X-ray and CT examination were performed 7 d and 1,3,6,12 months after operation to evaluate the internal fixation and bone grafting fusion.Results There's no occurrence of cervical spinal cord and vertebral artery injury.Screws were successfully implanted.1 case's incision occurred infiltration liquid 3 days after operation.For allograft rejection consideration,we changed the plan,and the infiltrating stopped 5 days later.Other incisions were all primary healing.VAS scores were 1.01±0.89 after operation,which was significantly reduced.X-ray showed good recovery of cervical sequence after the operation,and CT revealed 1 patient with atlas pedicle screw in the medial cortex damage.Spinal canal was without infringing,and the other screw positions were normal.6 months after operation,X-ray or CT examination both showed bony fusion.Conclusion Atlantoaxial pedicle screw fixation combined with unilateral axial spinous process and lamina screws and allograft bone graft fusion for the treatment of atlantoaxial instability can observe good clinical effects.

Escherichia coli AFP111 is a spontaneous mutant with mutations in the glucose specific phosphotransferase system (ptsG) in NZN111 (delta pflAB deltaldhA). In AFP111, conversion of xylose to succinic acid generates 1.67 molecule of ATP per xylose. However, the strain needs 2.67 molecule ATP for xylose metabolism. Therefore, AFP111 cannot use xylose due to insufficient ATP under anaerobic condition. Through an atmospheric and room temperature plasma (ARTP) jet, we got a mutant strain named DC111 that could use xylose under anaerobic condition in M9 medium to produce succinic acid. After 72 h, DC111 consumed 10.52 g/L xylose to produce 6.46 g/L succinic acid, and the yield was 0.78 mol/mol. Furthermore, the reaction catalyzed by the ATP-generating PEP-carboxykinase (PCK) was enhanced. The specific activity of PCK was 19.33-fold higher in DC111 than that in AFP111, which made the strain have enough ATP to converse xylose to succinic acid.
Atmosphere ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Industrial Microbiology ; Metabolic Engineering ; Mutation ; Plasma Gases ; pharmacology ; Succinic Acid ; metabolism ; Temperature ; Xylose ; metabolism

During the anaerobic fermentation by Escherichia coli AFP111 for succinic acid production, the viable cell concentration and productivity were decreased with the raising of succinic acid concentration. In order to restore cellular succinic acid productivity and prolong fermentation time, we collected strains and refreshed medium for repetitive succinic acid production. However, productivity is lower than that in the anaerobic fermentation before reusing strains. To enhance the productivity, strains were aerobically cultivated for 3 h in pure water before anaerobic fermentation. The activities of key enzymes were enhanced for better performance in producing succinic acid at anaerobic stage. After three rounds of repetitive fermentations, succinic acid concentration and yield reached to 56.50 g/L and 90% respectively. The succinic acid productivity was 0.81 g/(L x h), which was 13% higher than the repetitive fermentations without aerobic activation of the strains.
Aerobiosis ; Anaerobiosis ; Culture Media ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Genetic Engineering ; Glucose ; metabolism ; Industrial Microbiology ; Succinic Acid ; metabolism

Escherichia coli BA002, in which the ldhA and pflB genes are deleted, cannot utilize glucose anaerobically due to the inability to regenerate NAD+. To restore glucose utilization, overexpression of nicotinic acid phosphoribosyltransferase (NAPRTase) encoded by the pncB gene, a rate-limiting enzyme of NAD(H) synthesis pathway, resulted in a significant increase in cell mass and succinate production under anaerobic conditions. However, a high concentration of pyruvate was accumulated. Thus, co-expression of NAPRTase and the heterologous pyruvate carboxylase (PYC) of Lactococcus lactis subsp. cremoris NZ9000 in recombinant E. coli BA016 was investigated. Results in 3 L fermentor showed that OD600 is 4.64 and BA016 consumed 35.00 g/L glucose and produced 25.09 g/L succinate after 112 h under anaerobic conditions. Overexpression of pncB and pyc in BA016, the accumulation of pyruvic acid was further decreased, and the formation of succinic acid was further increased.
Anaerobiosis ; Escherichia coli ; enzymology ; genetics ; metabolism ; Fermentation ; Genetic Engineering ; Glucose ; metabolism ; Industrial Microbiology ; Lactococcus lactis ; enzymology ; NAD ; metabolism ; Pentosyltransferases ; biosynthesis ; genetics ; Pyruvate Carboxylase ; biosynthesis ; genetics ; Succinic Acid ; metabolism

Objective To study the incidence of ventilator-associated pneumonia(VAP)and antimicrobial resistance of pathogens in an intensive care unit(ICU).Methods The occurrence of VAP in hospitalized patients with mechan-ical ventilation>48 hours between January 2011 and December 2012 were investigated,species and antimicrobial re-sistance of pathogens causing early onset-VAP (E-VAP,mechanical ventilation≤4 d)and late-onset VAP(L-VAP, mechanical ventilation>4 d)were compared.Results A total of 1 76 patients were investigated,incidence of VAP was 44.32% (78 cases);With the prolongation of mechical ventilation,incidence of VAP increased gradually (χ2=52.561,P<0.001).The incidence of L-VAP was significantly higher than E-VAP (58.33% [70/120]vs 14.29%[8/56])(χ2= 30.02,P<0.001).A total of 178 pathogens were isolated,gram-negative bacteria,gram-positive bac-teria and fungi were 104(58.43% ),46(25.84% ),and 28(15.73% )isolates respectively;97(54.49% )multidrug-resistance/pandrug resistance organisms (MDRO)were isolated. MDRO isolation rate in L-VAP patients was high-er than E-VAP patients([58.86% ,n= 93]vs [20.00% ,n= 4]),resistance rate of major pathogens causing L-VAP was significantly higher than E-VAP patients(allP<0.05).Fungi infection only occurred in L-VAP patients,the total antimicrobial resistance rate was 12.14% .Conclusion The prolongation of mechanical ventilation can increase the incidence of VAP,and resistance rate of pathogen in L-VAP is high.

In order to provide anatomical basis for transoral approach (TOA) in dealing with the ventro lesions of craniocervical junction, and the design and application of artificial atlanto-odontoid joint, microsurgical dissecting was performed on 8 fresh craniocervical specimens layer by layer through transoropharyngeal approach. The stratification of posterior pharyngeal wall, course of vertebral artery, adjacent relationship of atlas and axis and correlative anatomical parameters of replacement of artificial atlanto-odontoid joint were observed. Besides, 32 sets of atlanto-axial joint in adults' fresh bony specimens were measured with a digital caliper and a goniometer, including the width of bony window of anterior arch of atlas, the width of bony window of axis vertebra, the distance between superior and inferior two atlas screw inserting points, the distance between two axis screw inserting points etc. It was found that the width of atlas and axis which could be exposed were 40.2+/-3.5 mm and 39.3+/-3.7 mm respectively. The width and height of posterior pharyngeal wall which could be exposed were 40.1+/-5.2 mm and 50.2+/-4.6 mm respectively. The distance between superior and inferior two atlas screw inserting points was 28.0+/-2.9 mm and 24.0+/-3.5 mm respectively, and the distance of bilateral axis screw inserting points was 18.0+/-1.2 mm. The operative exposure position through TOA ranged from inferior part of the clivus to the superior part of the C3 vertebral body. Posterior pharyngeal wall consisted of 5 layers and two interspaces: mucosa, submucosa, superficial muscular layer, anterior fascia of vertebrae, anterior muscular layer of vertebrae and posterior interspace of pharynx, anterior interspace of vertebrae. This study revealed that it had the advantages of short operative distance, good exposure and sufficient decompression in dealing with the ventro lesions from the upper cervical to the lower clivus through the TOA. The replacement of artificial atlanto-odontoid joint is suitable and feasible. The design of artificial atlanto-odontoid joint should be based on the above data.
Atlanto-Axial Joint/*anatomy & histology ; Atlanto-Axial Joint/*surgery ; Bone Plates ; Bone Screws ; Cadaver ; Cervical Vertebrae/*anatomy & histology ; Cervical Vertebrae/surgery ; Equipment Design ; Internal Fixators ; Joint Prosthesis ; Models, Anatomic ; Odontoid Process/*surgery ; Prosthesis Design

Objective To compare the clinical efficacy and fusion rate of unilateral and bilateral C1 and C2 pedicle screw fixation of unstable Jefferson fractures.Methods This retrospective casecontrol study enrolled 22 patients with unstable Jefferson fractures admitted between April 2012 and May 2015.There were 18 males and four females,with the mean age of 52.9 years (range,35-67 years).Mean preoperative visual analogue scale (VAS) was 6.09 points (range,4-8 points).According to the American spinal injury association (ASIA) classification,two patients were rated grade D and one patient grade C.Mean Japanese orthopedic association (JOA) score was 12.3 points.Bilateral C1 and C2 pedicle screw fixation was performed for 15 patients (bilateral group).Advantage side unilateral C1 and C2 pedicle screw fixation was performed for seven patients with extremely unstable fracture or narrow pedicle (unilateral group).Operation time,blood loss and surgical complications were recorded.VAS was used to evaluate the improvement of neck pain after operation.ASIA classification and JOA score were used to assess nerve function recovery.Atlanto-dental interval (ADI),srew position and bone fusion were evaluated after operation.Results All patients successfully completed the operation.Operation time was (119.5 ±21.2)min,and blood loss was (280.1 ±83.1)ml.A total of 74 screws were placed and CT scan showed satisfactory position of the screws.No complications were noted either during the operation or after surgery.All patients were followed up for mean 20.7 months (range,13-33 months).VAS was improved in both groups after operation (P ＜ 0.01),and there was no significant difference between the two groups (P ＞ 0.05).Two patients with ASIA grade D in bilateral group were improved to ASIA grade E after operation.One patient with ASIA grade C in unilateral group was improved to ASIA grade D after operation.JOA score increased to mean 15.7 points at last follow-up.ADI were decreased in both groups after operation(P ＜0.05),but there was no significant difference between the two groups (P ＞ 0.05).All patients had bony fusion 6 months after operation,with similar fusion rate between the two groups (P ＞ 0.05).Conclusion Advantage side unilateral screw fixation can be used for the patients with bilateral C1 and C2 pedicle screw fixation failure,for the technique can improve cervical pain and provide relatively high stability and fusion rate.