BACKGROUND: Neural stem cells are always derived from fetal rats and adult rats, and it is complex to isolate the cells by cell culture.OBJECTIVE: To study a convenient and effective method for the isolation and the culture of neural stem cells in neonatal rats.DESIGN, TIME AND SETTING: An observation study based on cells was carried out in the Chongqing Medical University (Chongqing, China) from October 2006 to March 2007.MATERIALS: Wistar neonatal rats of 1-3 days old.METHODS: Subsequent to trypsin digestion, primary culture of the cells was performed in serum-free suspension culture medium. Then the cells were induced to incubate in DMEM/F12 containing 0.10 volume fraction of fetal bovine serum. MAIN OUTCOME MEASURES: Phase contrast microscopy was employed to observe the growth of neural stem cells and the morphology of the differentiated cells. Neural stem cells and the differentiated neurons were identified using indirect immunofluorescence cytochemistry, as well as expression of gilal fibrillary acidic protein. Moreover, the proliferation of the BrdU-labeled neural stem cells was also investigated.RESULTS: The neural stem cells isolated from neonatal rat brains had the potential of serial passage and proliferation, besides, they express neuroepithelial stem cell protein (nestin) and differentiate into neurons, astrocytes and oligodendrocytes.CONCLUSION: Neural stem cells can be harvested from neonatal rat brains at a large scale, and they maintain their undifferentiated features and have the capacity of self-renewal and pluripotentiality.

Objective To investigate the effects of intrathecal (IT) gabapentin on the analgesic efficacy of morphine in a rat model of incisional pain. Methods Forty-eight male SD rats in which IT catheters were successly inserted according to the method described by Yaksh were randomly divided into 6 groups ( n = 8 each): Ⅰ sham operation group, Ⅱ incisional pain group, Ⅲ GBP 50 μg group, Ⅳ morphine 2.5 μg group, Ⅴ morphine 5 μg group, and Ⅵ morphine 2.5 μg + gabapentin 50 μg group. In group Ⅰ , IT artificial cerebrospinal fluid (ACSF)10 μl was injected and then 1.4% isoflurane was inhaled for 5 min. IT ACSF 10 μl, gabapentin 50μg and morphine 2.5 and 5 μg were injected 30 min before the establishement of the model in group Ⅱ -Ⅴ respectively. Paw withdrawl threshold (PWT) to mechanical stimulation and paw withdrawal latency (PWL) to a thermal nociceptive stimulus were measured at 2 h after the establishement of the model. Results Compared with group Ⅰ , MWT was significantly decreased and TWL was significantly shortened in group Ⅱ , Ⅲ and Ⅳ ( P ＜ 0.05), but no significant change in MWT and TWL was found in group Ⅴ and Ⅵ (P＞0.05). Compared with group Ⅱ , MWT was significantly increased and TWL was significantly prolonged in group Ⅴ and Ⅵ (P ＜ 0.05), but no significant change in MWT and TWL was found in group Ⅲ and Ⅳ/ ( P ＞ 0.05). MWT was significantly decreased and TWL was significantly shortened in group Ⅲ and Ⅳ compared with group Ⅵ ( P ＜ 0.05). Conclusion IT gabapentin enhances the analgesic efficacy of morphine in a rat model of incisional pain.

Objective To explore the early diagnosis, option of the surgical procedures, preven-tion and treatment of the complications in patients with pancreatic trauma.Methods All patients with pancreatic trauma during the past 20 years were studied retrospectively in respect of sex, age, cause of injury, grade by AAST, style of operation, therapeutic efficacy, complications and factors for death etc.Statistical analysis was made with Chi-square test.Results In all 148 cases of the present series, 132 underwent surgical interventions including simple suture or external drainage alone, distal pancre-atectomy, distal pancreaticojejunostomy or other internal drainage, diverticularization, Whipple's pro-cedure, and Damage Control Surgery (DCS) etc.Postoperative morbidity was 27.83% with a signifi-cant difference of the incidence of pancreatic fistula between pancreas grade Ⅲ-Ⅴ injuries and grade Ⅰ-Ⅱ(P＜0.01).The mortality rate was 11.49%.The cause of the deaths was mainly massive bleed-ing due to severe associated injuries(76.47%).Among various grades, the difference of the mortality was not significant (P＞0.05).Conclusion To improve the survival rate, it is important to control massive hemorrhage from associated injuries precedes dealing with pancreas trauma.Selection of surgi-cal procedures should be based on whether the main duct is injured.The removing of devitalized tis-sue, adequate external and internal drainage are essential for treatment of pancreatic injuries.Early recognition of pancreatic injury and correct choice of surgical procedures may obviously decrease the in-cidence of postoperative complications.

Objective To investigate the effect of pemetrexed on the immune function,quality of life and survival time of patients with malignant pleural effusion.Methods Eighty cases patients with non-small cell lung cancer from July 2013 to October 2015 in the People′s Hospital of Hunan Province were complicated with pleural effusion,and randomly divided into observation group and control group,40 cases in each group.The control group were used chemotherapy with GP(gemcitabine plus cisplatin) and the central venous catheter was placed in the thoracic cavity.The observation group was treated with retrograde injection of pemetrexed catheter on the basis of the control group.The changes of immunoglobulin and inflammatory cytokines were compared between the two groups,and the quality of life and the 1 year survival rate of the two groups were statistically analyzed.Results The IgM,IgG and IgA of the observation group after the intervention were higher than those in control group((1.65±0.03) mg/L vs.(1.31±0.02) mg/L,(9.55±0.12) mg/L vs.(8.82±0.10) mg/L,(3.99±0.20) mg/L vs.(1.81±0.13) mg/L,t=59.640,29.557,57.800,P<0.05),the TNFα-,IL-1,hs-CRP levels of the observation group after the intervention were lower than the control group((12.0±0.2) μg/L vs.(18.1±0.5) μg/L,(0.60±0.1) mg/L vs.(0.92±0.2) mg/L,(10.3±1.0) mg/L vs.(31.3±2.0) mg/L,t=71.641,9.051,59.397,P<0.05),and quality of life score after the intervention than before((81.5±2.3) points vs.(81.6±2.3) points,(35.3±1.1) points vs.(56.6±1.7) points,t=114.608,55.823,P<0.05).Quality of life of the observation group was better than the control group(t=66.530,P<0.01),1 year survival rate was higher than the control group(25.0%(10/40) vs.50.0%(20/40),U=5.903,P<0.05).Conclusion Local chemotherapy with pemetrexed could significantly improve the immune function of patients with malignant pleural effusion,reduce the inflammatory response,improve the quality of life and prolong the survival time of patients with malignant pleural effusion.

Objective To study the application of pressure regulated volume control ventilation in respiratory support after liver transplantation.Methods Twenty patients underwent liver transplantation were randomly averagely divided into two groups: pressure regulated vlume control ventilation(PRVCV) group and volume control(VC) group.The parameters of respiratory mechanics,hemodynamics and blood gas analysis of patients in two groups were compared,such as oxygen delivery(DO2),oxygen consumption(VO2),oxygen incepation ratio(O2ER),arteriovenous oxygen content difference(C(a-v)O2),cardiac output(CO),mean arterial pressure(mABP),mean pulmonary arterial pressure(mPAP),alveolar-arterial PO2 difference(P(A-a)O2),gas exchange index(PaO2/FiO2),ratio of shunted blood to total perfusion(Qs/Qt),peak inspiratory pressure(PIP) and mean airway pressure(mAP).Results The P(A-a)O2 and Qs/Qt were significantly decreased in PRVCV group than those in VC group(P(A-a)O2:(101.42?28.07) mm Hg vs.(136.76?39.13) mm Hg;Qs/Qt:(1.78?0.86)% vs.(3.28?0.99)%),P

Objective To construct the eukaryotic expression plasmid of pEGFPC1uPAR gene and explore the effect on the proliferation and invasion ability of Pam 212 cells. Methods The human uPAR cDNA was cloned by PCR, and inserted into the eukaryotic expression plasmid pEGFPC1. After identification of sequencing, the reconstructive plasmid was transformed transiently into Pam 212 cells, then the cell growth and the invasion ability were evaluated. Results The reconstructive plasmid of pEGFPC1uPAR was validated by sequencing. The reconstructive plasmid can promote the growth of Pam 212 cells and enhance the invasion ability. Conclusion The pEGFPC1uPAR plasmid was constructed successfully and uPAR was confirmed to promote the growth and the invasion ability of Pam 212 cells, which lay the foundation for further studies of uPAR in vivo.

Objective To summarize and report the experiences of surgical treatment of infective endocarditis. Methods From March 1995 to March 2004, surgical operations were performed in 25 patients with infective endocarditis, 9 of who received emergency surgery and 16 received selective operation. Replacement of mitral valve was performed in 3, aortic valve replacement in 13, mitral and aortic valve replacement in 8, Bentall procedure in 1.Associated cardiac lesions included patent ductus arterioles in 1, ventricular septal defect in 2, rupture of Valsalvaps sinus in 1. Results There was one postoperative death, which gave an overall hospital mortality of 4%. Twenty-four patients were cured and improved. Conclusion Surgical operation should be the preferred mode of surgical correction for infective endocarditis. The timing of surgery and proper perioperative management are key factors for the successful treatment of infective endocarditis.

Objective To study the biological exposure limit in bone metabolism damage caused by coexposure to fluorine and arsenic from coal burning in exposed population.Methods One hundred and ninety-eight cases of fluoride and arsenic co-exposed people from Liuchang village,Qinzhen city,Guizhou province were enrolled in the study.Urinary fluorine (UF),urinary arsenic (UAs),urinary hydroxyproline (UHYP),ross-linked Ntelopeptides of type Ⅰ collagen(UNTX) and bone strength index(STI) were detected.BMDS Version 2.1 software was used to calculate UF,UAs benchmark dose (BMD) and its lower confidence limit (BMDL) on the damage of bone metabolism caused by co-exposure to fluorine and arsenic from coal burning.Results The BMD and BMDL range of UF caused by co-exposure to fluorine and arsenic from coal burning were 0.68-1.35 mg/g Cr,0.57-1.11 mg/g Cr.The BMD and BMDL range of UAs caused by co-exposure to fluorine and arsenic from coal burning were 8.36-18.77 μg/g Cr,7.12-15.40 μg/g Cr.Conclusion The biological exposure limits of UF and UAs for bone metabolism toxicity are proposed as 0.57 mg/g Cr and 7.12 μg/g Cr in co-exposure to fluoride and arsenic from coal burning,respectively.

Objective To detect and explore the expression of ARD1 and its clinical significance in the nasopharyngeal in-flammatory tissue ,nasopharyngeal carcinoma group and its subgroups .Methods Expression of ARD1 in nasopharyngeal carcinoma (56 cases) and nasopharyngeal inflammatory tissue (20 cases) were detected by immunohistochemical staining SP ,the correlation between the expression of ARD1 and age ,gender ,histological grade ,TNM clinical stage and tumor metastasis were analysed . Results The positive expression rate of ARD1 were 10 .00% (2/20) ,55 .35% (31/56) in the nasopharyngeal inflammatory tissue and nasopharyngeal carcinoma ,respectively .The expression level of ARD1 in nasopharyngeal carcinoma was significantly higher than in the nasopharyngeal inflammatory tissue ,the difference was significant (P< 0 .05) ;expression of ARD1 in the nasopharynge-al carcinoma was correlated with the histological grade of nasopharyngeal carcinoma(P< 0 .05) and the expression was increased in poor differenciation tissue .But there was no statistical difference between the expression of ARD1 and the patient′s age ,gender , TNM clinical stage ,tumor metastasis(P> 0 .05) .Conclusion The expression of ARD1 is high in nasopharyngeal carcinoma ,and has a closely correlation with diffferentiation level of tumor ,which suggested that ARD1 may be involved in the the occurrence and development of nasopharyngeal carcinoma .However ,further research needs to be done for its mechanism in the nasopharyngeal car-cinoma .

Objective To evaluate the safety and feasibility and clinical effectiveness of living relative donor kidney transplantation(LDKT)and summarize its clinical experience. Methods The clinical data of 30 cases of LDKT were retrospectively analyzed.Except for 2 cases being donated by spouse,the others were donated by blood relative donors.6 cases shared two haplotypes,and 22 cases shared one haplotype,and one case 4 mismatched,and 1 fully mismatched.All donors underwent open nephroectomy,in which 7 cases donated right kidneys and 23 donated left kidneys.In 30 cases of recipients,1 case received cadaver donor kidney transplantation and lost her allograft because of superacute rejection.Triple-combined immunosuppressive protocols consisted of calcineurin inhibitor (CNI),mycophenolate mofetil(MMF)or azathioprine(AZa)and steroid. Results All donors'hospital stay was 7 to 10 days postoperatively without any surgical complications. All donors kept their normal kidney function within 3 to 6 months'follow-up.Except for 1 case of death because of lung in fection,29 cases of recipients survived,in which 28 cases kept their normal function kidney within 1 to 4 years of follow-up and 1 case occurred chronic allograft nephropathy after one year.Except one case of DGF,29 cases of recipients retained their normal kidney function in 3 to 5 days postoperatively.Rejection episodes occurred in 4 cases,of which 3 cases were reversed by methylprednisone and 1 case by antithymocyte globulin(ATG)and Tacrolimus.Pneumonia developed in 3 cases,of which 2 cases were cured and 1 case failed.Hematoma was found around allograft in 1 case and wag surgi cally removed.Urinary leakage was happened in 2 cases of recepients and were cured by conservative treatment. Conclusions LDKT is safe and feasible with good long-term results and more advantages such as optimal HLA matches and less ischemia time and lower acute rejection,low-dose immunosup pressants.