Purpose　The aim is to optimize the liquid culture conditions of dihydropyrimidinase producing strain Pseudomonas putida 9801. Methods　Plackett-Burman design and spherical symmetric desig n were used.Results　Optimum conditions for dihydropyrimidinas e formation of Pseudomonas putida 9801 were defined:yeast extract 2.39%, Glu cose 1.81%,Uracil 0.06%,K2HPO4*3H2O 0.2%, MgCl2*6H2O 0.05%and NaCl 0 .3%,when the strain was cultured at 32℃ for 10 h,about 3.02 units/ml of hydanto inase was obtained. This value was quite consistent with the theory value(2.91 u nits/ml).Conclusion　The liquid culture conditions of dihydrop yrimidinase producing strain were optimized.

AIM　The purpose is to construct D-hydantoinase genetic engineering strain for the purpose of the industrial production of D-p-hydroxyphenylglycine. METHODS　D-hydantoinase gene was created from Pseudomonas putida 9801 by PCR technique and inserted into pMD18-T vector. The recombinant plasmid was transformed into several Escherichia coli strains. The positive transformants with D-hydantoinase activity were obtained by the two step screening, digoxigenin DNA labeling in situ hybridization and D-hydantoinase activity assay. RESULTS　The D-hydantoinase activity of the genetic engineering strain E.coli BL21/pMD-dht was 1700 U*L-1 and increased as high as 8 times compared with those of wild-type strain Pseudomonas putida 9801. The subunit molecular weight of recombinant D-hydantoinase was about 53 kDa measured by SDS-PAGE. The amount of the recombinant D-hydantoinase was about 20 percent of total bacterial soluble proteins. CONCLUSION　The genetic engineering strain E.coli BL21/pMD-dht possesses the initial industrial production prospects.

Objective To investigate the optimal operative approach for esophageal and cardiac carcinoma,in order to obtain favorable exposure and to diminish trauma.Methods A method of median-rib-Section were performed in 207 cases of esophageal and cardiac carcinoma:the 6th and 7th ribs were cut intermediately,then we peeled and cut the rib periosteum.stretching the costal interspace with a prop until the exposure was optimal.Items including traumatic degree,exposure extend and conditions of thoracic cavity closure were compared with that of traditional antemposterior-rib-section method.Results The exposure result was similar in the two methods,but the traumatic de-grce WaS less when anteroposterior-rib·section was applied,due to its avoiding cutting intercostal muscles which caused more blood loss.The median-rib-section method led to more satisfactory anatomy paratope and external appearance,less trawma and incision pain.Conclusion The method of median-rib-section is worthy of generalization because of its satisfactory exposure,less trauma,better paratope and absence of postoperative costal malformation.

Objective To examine the practicability and security of the LigaSureTM vessel-sealing system in radical gastrcctomy.Methods Seventy patients of stage Ⅰ～Ⅲ gastric cancer undergone radical gastrectomy were analyzed retrospectively.Thirty-five radical gastrectomy were performed with LigaSureTM vessel-sealing system,the other 35 eases were performed without LigaSureTM.The operative duration,volume of intraoperative hemorrhage,and postoperative complications were analyzed.Results There were significant difference in both operative duration[(159.9±24.7)min vs(172.6±23.7)min,P＜0.05]and volume of intraoperative hemorrhage[(335.7±40.3)ml vs(371.7±43.2)ml,P＜0.01].There was no significant difference in the postoperative complications between LigaSureTM group and conventional method group.All patients recovered without severe postoperative complications.Conclusion Radical resection of gastric cancer using LigaSureTM instead of conventional hand tie method appears to he shorten operative duration,decreased volume of intraoperative hemorrhage.And the technique is convenient and safe.

Objective To explore the feasibility,necessity and the skill of total mesoesophageal excision (TME ) during thoracoscopy combined with laparoscopy in radical resection of esophageal carcinoma.Methods 69 patients with esophageal carcinoma were divided into the TME group(40 cases)and the thoracotomy with triple incisions group(29 cases)according to the admission sequence.The operation time,intraoperative blood loss,total lymph nodes removed,postoperative complication rate and disease -free survival were compared between the two groups.Results The operation time of TME group was (182.85 ±26.73)min,which was significantly shorter than (295.71 ±19.50)min of the thoracotomy group (t=-19.301,P<0.001).The intraoperative blood loss in TME group was (86.43 ±59.34)mL,which was significantly less than (163.47 ±58.82)mL in the thoracotomy group (t=-5.342,P<0.001 ).No significant differences were detected between the two groups in total lymph nodes removed and incidence rate of postoperative complication (all P>0.05 ).The disease-free survival period in TME group was (14.78 ±2.14)months,which in the thoracotomy group was (13.10 ±4.09)months,the difference was significant (t=2.200,P<0.05).Conclusion TME is safe and feasible during thoracoscopy combined with laparos-copy in radical resection of esophageal carcinoma.TME is better in improving the regional control in esophageal carci-noma.

In this paper, the biological compatibilities of FGF/Collagen compound sponges are evaluated. Intracutaneous stimulation test and shortterm muscle embedding test of FGF/Collagen compound sponges made of different materials are performed. Experimental data are analyzed and evaluated according to the standard. The two sponges get marks of 0 in acute eye stimulation test and intracutaneous stimulation test, thus they are not stimulating to tissues. In shortterm muscle embedding test, they don't lead to inflammation reactions and can be degraded and absorbed in short term. FGF/Collagen compound sponge proves good biological compatibility and has a cheerful prospect in medicine.

The personnel training mode of school-enterprise cooperation, interaction of study and work is the inevitable trend of senior vocational education reform and development at present. The Changsha Social College cooperates closely with rehabilitation agencies and implements the personnel training mode of the “love to help people with disabilities, interaction of study and work, integration of courses and certificate” for training professionals of rehabilitation therapy, has gained some experience and achievements.

Objective The purpose of this study was to develop a database program based on Microsoft Access 2003 for patients with breast cancer to save and manage clinical data.This makes it easier to analyze data of the database.Methods A total of 1177 cases with breast cancer who were performed surgical treatment from December 1998 to June 2007,the clinical data collection which included regular fouow-up data of these patients after therapy,the database was designed by using Microsoft Access 2003 included the creation of tables,forms and queries according to the therapy guidelines for breast cancer.Results The clinical data for patients with breast cancer can be stored in an Access database that is both user-friendly with reliable data administration and that can be shared in network,and it is capable of recording regular updated data.Conclusion This program will not only convenient for researchers to statistic and analyze their outcome data,but also for minimizing data entry errors and reducing the time spent on data query.

Objective To evaluate the efficacy of zero ischemia index (ZⅡ) in predicting the complexity and perioperative outcomes of off-clamp nephron-sparing surgery (NSS).Methods The patients between June 2016 and June 2017 in our institution who underwent off-clamp NSS were prospectively evaluated.ZⅡ was defined as the product of the tumor diameter and depth within renal parenchyma.The ZⅡ ＞6 defined as higher risk while ZⅡ ≤ 6 defined as low risk.The operating time,estimated blood loss,hospital stay,drainage,and complication rate were analyzed.Results There were 35 males and 10 females with average age of 42 y(range 23-76y).Mean tumor size was 2.4 cm (range 0.8-4.2 cm).Mini-flank approach open NSS was performed in 33 cases and laparoscopic NSS was performed in 12 cases.Off-clamp NSS was successfully performed in 44 patients except for renal artery occlusion in 1 case.Mean operative time was (95.0± 17.5) min (range 50-150 min);The average estimated blood loss was (152.4 ± 134.2) ml (range 20-600 ml);Mean postoperative drainage was (97.3 ± 59.7) ml (range 50-300 md);Mean postoperative hospital stay was (6.1 ± 1.3) d (range 5-8 d).Not severe post operative complication was observed.There were 37 patients in low risk group and 8 patients in high risk group.Operating time was significantly longer in high risk group [(118.8 ± 14.6 min) vs.(89.9±13.4) min,P＜0.01].EBL [(375.0±158.1) ml vs.(104.3 ±61.4) ml,P＜0.01] and drainage [(161.2±91.3)ml vs.(83.5 ±40.4)ml,P ＜ 0.01] were also significantly higher in high risk group.But there was no significant difference in hospital stay between two groups.The postoperative pathology indicated that 35 cases of clear cell carcinomas,2 cases of chromophobe renal cell carcinomas,one case of papillary carcinoma and seven cases of angiomyolipomas.Conclusions The ZⅡ is a novel and effective measurable criterion which can help predict the risk of perioperative outcomes of off-clamp NSS.ZⅡ =6 is established as a preliminary threshold for patient selection of off-clamp NSS.

Objective:To evaluate the role of extracellular signal-regulated kinase (ERK)1/2 in glutamate-induced ferroptosis in PC12 cells.Methods:PC12 cells were divided into 6 groups ( n=21 each) using a random number table method: control group (C group), glutamategroup (Glu group), glutamate+ ERK1/2 over-expression group (Glu+ ERK1/2-OE group), glutamate+ ERK1/2 plasmid empty vector group (Glu+ Vec group), glutamate+ ERK1/2 knockdown group (Glu+ si-ERK1/2 group)and glutamate+ ERK1/2 SiRNA negative control group (Glu+ si-NC group). Cells were treated with glutamate at a final concentration of 6 mmol/L for 72 h in Glu group and with the equal volume of PBS buffer for 72 h in C group. Glu+ ERK1/2-OE group was transfected with ERK1/2 overexpression plasmid, Glu+ Vec group was transfected with plasmid empty vector, and Glu+ si-ERK1/2 group was transfected with ERK1/2 siRNA, Glu+ si-NC group was transfected with siRNA negative control for 48 h, and then glutamate at a final concentration of 6 mmol/L was added and cells were treated for 72 h. The cell viability, lactic dehydrogenase (LDH)activity and contents of glutathione (GSH), ferrous ions and malondialdehyde (MDA) were measured by enzyme-linked immunosorbent assay. Mitochondrial membrane potential (MMP) and lipid reactive oxygen species (Lip-ROS) were measured by flow cytometry. Results:Compared with C group, the cell viability, GSH content and MMP were significantly decreased, and the LDH activity, ferrous ions content, MDA content and Lip-ROS levels were increased in Glu group ( P<0.05). Compared with Glu+ Vec group, the cell viability, GSH content and MMP were significantly increased, and the activity of LDH, contents of ferrous ions and MDA, and Lip-ROS levels were decreased in Glu+ ERK1/2-OE group( P<0.05). Compared with Glu+ si-NC group, the cell viability, GSH content and MMP were significantly decreased, and the LDH activity, contents of ferrous ions and MDA, and Lip-ROS level were increased in Glu+ si-ERK1/2 group ( P<0.05). Conclusions:ERK1/2 is involved in glutamate-induced ferroptosis in PC12 cells.