Background The deficiency of limbus stem cells will cause various kinds of the disorder of eye surface,in serious condition,it will lead to corneal conjunctivalization,chronic inflammation,lasting deficiency of corneal epithelium and corneal opacity accompanying long-term visual loss.Objective The aim of this study was to investigate how to establish and evaluate the successful model of total limbal stem cell deficiency (LSCD).Methods Twenty Japanese white rabbits aged 3-5 months were selected.The limbus corneal epithelium was removed from 1 mm inner to 2 mm outer of the limbus 100-150 μm in deep,and the total central epithelium and shallow stroma were simultaneously removed in the left eyes using scalpel.The experimental eyes were examined by slitlamp 2,3,4,5 weeks after operation,and the inflammatory score was performed according to corneal opacity,corneal neovascularization and corneal flurescein dye using international universal rating criteria of LSCD model.Cornea tissue was obtained in various time points mentioned above to examine the structural change and goblet cells.The expression of cytokeratin 3(CK3) in the cornea was detected using indirect immuno-fluorescence staining.Fisher Exact Probability method was used to compare the successful rate of models in different time points.Results The successful rate of models rate was 12.5％,62.5％,81.3％ and 87.5％,respectively 2,3,4,5 weeks after operation,showing a significant difference between the 3-week group and 2-week group (P =0.009).However,no significant differences were found in the model successful rate between the 3-week group and the 4-week group,the 4-week group and the 5-week group (P =0.465,0.200).There was a little difference between the 3-week group and the 5-week group(P=0.049).The mean time of achieving successful model was(3.21 ±0.80) weeks.The result of hematoxylin & eosin staining showed the edema of cornea stroma,a mass of inflammatory cells infiltration and new vessels.Goblet cells could be found by Periodic acid-Schiff staining 5 weeks after operation.Expression of CK3 was absent in the corneas of successful model eyes.Conclusions The LSCD model can be successfully established by cutting off the epithelium of limbus and central cornea,and the average time of model formation is about 3.2 weeks.

Objective To determine the proper dosage of fentanyl for open heart surgery performed under cardiopulmonary bypass (CPB) without aortic cross-clamping and cardioplegia.Methods Twenty-seven ASA Ⅰ -H patients (10 male, 17 female) with fairly good cardiac function (NYHA Ⅰ - Ⅱ) scheduled for surgical repair of atrioseptal defect ( ASD) or ventricular septal defect (VSD) or mitral valve replacement (MVR) were studied. Age ranged from 18 to 44 years and body weight from 35 to 58 kg. The patients were randomly divided into three fentanyl dosage groups: group Ⅰ 10?kg ; group II 30 ?g kg-1 and group Ⅲ 50 ?g kg-1. Premedication consisted of intramuscular pethidine 1-2 mg kg-1 and scopolamine 0.05-0.06 mg kg-1 . Anesthesia was induced with midazolam 0.2 mg kg-1 and fentanyl 5 ?g kg-1 . Tracheal intubation was facilitated with vecuronium 0.15 mg g-1 . The patients were mechanically ventilated (Vr 8-10 ml kg-1 ,F 10-12 bpm,FiO2 100% ). The rest of the total dose of fentanyl (5 ?g kg-1 in group I , 25 ?g kg-1 in group II , 45 ?g kg-1 in group III) was infused after induction of anesthesia until the initiation of CPB, supplemented with inhalation of 0.6 % isoflurane. During CPB propofol was infused at 5 mg kg-1 h-1 . after discontinuation of CPB, again 0.6% isoflurane was inhaled until the end of surgery. Vecuronium 0.05 mg kg was given every 25-30 min during operation. EGG, HR, BP, CVP, SpO2, PET CO2 and body temperature (naso-pharyngeal and rectal) were continuously monitored during operation. Arterial blood samples were obtained before anesthesia (T0), 5 min after tracheal intubation (T, ) , immediately after thoracotomy (T2) , immediately before CPB (T3), 15 min after CPB was initiated (T4) , 10 min after termination of CPB (T5) and 5 min after chest was closed (T6) for blood gas analyses and determination of blood electrolytes and acid-base balance and blood concentrations of glucose, ACTH, angiotensin Ⅱ (A- Ⅱ ) and cortisol. Time of emergence from anesthesia and extubation were recorded.Results The demographic data, including age and body weight, CPB time and duration of surgery were comparable among the three groups. There were no significant changes in SpO2 , PETCO2 , body temperature, blood gases and electrolytes during operation in the three groups. MAP decreased significantly during CPB. The blood glucose, ACTH, A- Ⅱ and cortisol concentrations increased significantly during and after CPB as compared with the preanesthetic baseline (T0 ) ( P

Objective:To investigate the ethical significance of the extracorporeal circulation membrane oxy-genation ( ECMO) in the donation organ transplantation .Methods: Analyzing the data of ECMO protected to the organ in 13 donors after brain death , Accounting the rising costs which caused by ECMO and make the interview to the patients and family members .Results:In the period of ECMO flow , the hemodynamic of the DBD donors be-come stable gradually , the medications reduced significantly or stop , the function of organs was restored .There were 38 organs can be used for the transplantation which were proven by the pathological biopsy .Twenty six kid-neys were transplanted to 26 recipients and liver transplantation was performed in 12 recipients.All transplantations were successfully completed .Medical cost of this patients increase 5.3%, all of the family members and patients can accept the intervention of ECMO .Conclusion:ECMO is an effective method to protect and improve the utili-zation rate of the organ .the improvement of the related technical standards , legal, laws and ethics of staff will pro-mote to the development of organ transplantation .

Objective To compare clinical decision-making ability of nursing undergraduate students in different internship hospitals and provide a basis for improving the cultivation of clinical decision-making ability. Methods The instrument for measuring the clinical decision-making ability of nursing students was adopted to investigate nursing undergraduate students from four colleges and universities in Sichuan province who had completed 10～12 months clinical practice.Results Nursing undergraduate students who undertook clinical practice at affiliated hospitals of medical colleges and universities got higher scores than those at non-affiliated hospitals,and there was significant difference between the two groups(P<0.01).Conclusions The clinical nursing decision-making ability of the nursing undergraduate students who undertook clinical practice at affiliated hospitals of medical colleges and universities is better than that of those at non-affiliated hospitals.

Objective To construct a new recombinant immunotoxin expression vector by using human VEGF165 and a truncated pseudomonas exotoxin A ramification (PE38) gene, and explore the expression of the VEGF165-PE38 fusion protein in HEK293 cells. Methods VEGF165 was cloned by polymerase chain reaction (PCR). PE38 gene was gained from an vector plasmid pRB391 by restriction endonuclease digestion, and then inserted to the eukaryotic expression vector pIRES2-EGFP. After the eukaryotic recombinant vector pIRES2-VEGF165-PE38-EGFP was identified by restriction endonuclease digestion and sequence analysis, the vector was transfected into HEK293 cells by liposome protocol. RT-PCR and ELISA method was used to confirm the expression of the fusion gene in the HEK293 cells. Results Restriction endonuclease digestion and sequence analysis revealed the VEGF165-PE38 fusion gene was cloned into the eukaryotic expression plasmid vector pIRES2-EGFP successfully. The pIRES2-VEGF165-PE38-EGFP fusion gene could express in the HEK293 cells. Conclusion The result provide the basis for search of the targeted cytotoxic activity to tumor vascular endothelial cells and may have some potential value in clinical application.

ObjectiveTo define the maximum tolerated dose (MTD) of weekly cisplatin in concurrent chemoradiotherapy for Chinese cervical carcinoma.MethodsCervical carcinoma of stage ⅠB2- ⅣA were eligible for the study.PhaseⅠstudy was dose-escalation trial with 15 patients.All patients received whole pelvic radiotherapy with three dimentional conformal radiotherapy technique. Concurrent cisplatin started from the dose of 20 mg/m2 to 25 mg/m2,30 mg/m2,35 mg/m2,40 mg/m2 for the weekly schedule ( ≥3 patients per dose group) and the doses were steadily escalated to 40 mg/.m2.If the dose was increased to 40 mg/m2 without dose-limiting toxicity ( DLT),40 mg/m2 would be the maximum tolerated dose (MTD).According to the MTD dose from Phase Ⅰ study,we conducted phase Ⅱ clinical trial with 36 patients.ResultsIn Phase Ⅰ study,cisplatin dose was escalated to 40 mg/m2 and DLT had not been reached.Thirty-six patients in Phase Ⅱ study included 9 inpatients and 27 outpatients.All 9 inpatients completed 6 cycles of chemotherapy. In 27 outpatients,18 patients (66％) completed 6 cycles of chemotherapy,19 patients (70％) completed 5 cycles and 25 patients (92％) completed 4 cycles of chemotherapy.All patients completed radiotherapy.Major adverse effects were grade 1 and 2 gastrointestinal toxicities and neutropenia.ConclusionsWeekly 40 mg/m2 cisplatin concurrent with radiotherapy is well tolerated when given to Chinese patients with cervical carcinoma. For outpatients with poor performance status,the cisplatin dose needs to be reduced.

OBJECTIVE: The study was carried out on time limit of ACP and sperm positive-detected in mixed spot. METHODS: 600 vaginal swabs from living body after sexual intercourse were tested by routine and three-blindness methods. RESULTS: The longest time limit of ACP which was positively detected is 255 h, average time limit is 52 h, 41.3% of those samples was ACP-detected in positive degree (++). The longest time limit of sperm positively detected is 132 h, average time limit is 29 h. The number of sperms observed in 34.3% of those samples with positive result, is from 6 to 10/HP. CONCLUSION This study is helpful to forensic identification of sex cases.
Acid Phosphatase/analysis* ; Coitus ; Female ; Forensic Medicine ; Humans ; Male ; Spermatozoa ; Time Factors ; Vaginal Smears

Objective To compare the clinical decision-making ability of baccalaureate nursing students from different colleges and universities and provide a basis for these schools to strengthen the cultivation of clinical decision-making ability. Methods The Instrument for Measuring the Clinical Decision-making Ability of Nursing Students was adopted to investigate baccalaureate nursing students from four colleges and universities in Sichuan Province who had completed 10 ～ 12 month clinical practice. Results Differences in clinical decision-making ability of baccalaureate nursing students existed among the four schools of Sichuan Province.The highest score was ( 94. 06 ± 5.09 ) and the lowest score was ( 89. 01 ± 5.68 ). The score between universities showed statistical differences( P ＜0. 05 ). Conclusions The clinical nursing decision-making ability of baccalaureate nursing students of the key university in Sichuan Province, authorized directly by the Ministry of Health, is generally better than that of ordinary universities and colleges. Nursing education in the key university in Sichuan Province is more beneficial to the cultivation of clinical nursing decision-making ability than that in ordinary universities and colleges.

Objective To compare the critical thinking ability between the baccalaureate nursing students in the four-year nursing program and the five-year nursing program and provide a basis for these schools with different school systems to strengthen the cultivation of critical thinking ability. Methods The Critical Thinking Disposition Inventory-Chinese Version (CTDI-CV) was adopted to investigate baccalaureate nursing students from four colleges and universities in Sichuan Province who had completed 10 ～ 12 month clinical practice. Results The overall mean score of critical thinking ability of baccalaureate nursing students in Sichuan Province was (287.37 ± 32. 52). The overall mean scores of critical thinking ability of students in the four-year nursing program and the five-year nursing program were (295.32 ±29. 76) and (282.63 ±33.23 ) respectively and a difference existed between them ( P ＜ 0. 01 ). Except the dimension of self-confidence, the differences between the two programs showed a statistical significance in the other six dimensions (P ＜0. 05). Conclusions Differences in critical thinking ability exist between baccalaureate nursing students in different school systems and the nursing education of four-year nursing program is more beneficial to the cultivation of critical thinking ability than that of five-year nursing program.

Objective To investigate baccalaureate nursing students'subjective well-being before clinical practice and discuss the influential factors in order to help students build a positive state of mind to participate in the clinical practice.Methods The convenience sample consisted of 144 baccalaureate nursing students who would take part in clinical practice and the General Self-efficacy Scale (GSES) and General Well-being Schedule (GWB) were adopted.Results The baccalaureate nursing students' subjective well-being was generally at a little higher level than the intermediate.Among the dimensions,the scores of life satisfaction,positive affect and negative affect were (4.69 ± 0.61 ),(4.12 ± 1.13 ) and (2.44 ± 0.89 ),respectively.Among the students,those who were the only child in the family or who were excellent in their study had higher subjective well-beings,with ( 24.98 ± 5.64 ) and ( 24.44 ± 4.40 ),respectively.The main influential factors included self-efficacy,self-esteem and the number of children in the family.Conclusions Nursing educators and psychologists should attach importance to baccalaureate nursing students' subjective well-being before clinical practice.Self-efficacy and self-esteem can obviously predict the change of subjective well-being.