Objective To investigate the gene expression of GDNF mRNA on both ends of the severed sections of sciatic nerve in rats. Methods The specimen of sciatic nerve proximal and distal to the cut off site was taken respectively before and after sciatic nerve severance. The level of GDNF mRNA on both cutting sections was observed and compared by using of semi quantitative RT-PCR method with ? Actin as a inner consult before nad after sciatic nerve cut off. Results A minimum expression of GDNF mRNA was found on sciatic nerve before the operation, however, there was an increase of 20% at 24 hours in the distal section after sciatic nerve cut off, 60% at 7th day, 85% at 14th day and 90% at 28th day. While it decreased by 10%, 38%, 45% and 52% at 1st day, 7th day, 14th day and 28th day in the proximal section respectively. Conclusion The decreasing and increasing of GDNF mRNA level at the proximal and distal sciatic nerve section is the result of the disruption of “neuron axon target”axis after sciatic nerve cut off. This may provide a theory basis in treatment of SCI by use of the foreign GDNF.

0 05) While patients′ satisfactory rate was significantly higher in electrical stimulation group than in medical group( P

Baculovirus has many advantages as vectors for gene transfer. We demonstrated that recombinant baculovirus vectors expressing p35 (Ac-CMV-p35) and eGFP (Ac-CMV-GFP) could be transduced into human kidney 293 cells efficiently. The level of transgene expression was viral dose dependent and high-level expression of the target gene could be achieved under the heterogonous promoter. MTT assay suggested that both Ac-CMV-p35 and Ac-CMV-GFP did not have cytotoxic effect on human embryo kidney 293 cells. Cell growth curve showed the Ac-CMV-p35 and Ac- CMV-GFP transduced and non-transduced cells had similar proliferation rate, so baculovirus-mediated p35expression had no adverse effect on cell proliferation. In addition, baculovirus-mediated p35 gene expression protected human embryo kidney 293 cells against apoptosis induced by various apoptosis inducers such as Actinomycin D, UV or serum-free media. These results suggested that the baculovirus vector mediated p35 gene expression was functional and it could be widely used in molecular research and even gene therapy.

Objective To assess the effect of radiofrequency ablation ( RFA) combined with transcatheter hepatic arterial chemoemboli-zation ( TACE) and radiofrequency ablation alone for the treatment of early hepatocellular carcinoma ( HCC) with liver cirrhosis. Methods The data of all the patients that were given RFA+TACE (n=51) and RFA (n=53) treatments in our hospital from January 2008 to De-cember 2013 were analyzed. The demographic data,process of operation,postoperative complications,postoperative recovery and follow-up of the patients in two groups were carefully compared. Results The average follow-up time was (37. 6 ± 20. 7)months,38 cases were dead and 76 cases were survival. The overall survival rates of 1 years,3 years,5 years after operation were respectively 96. 5%,78. 7%,65. 9% in RFA+TACE group and 94. 3%,75. 6%,62. 3% in RFA group. Tumor-free survival rates of 1 years,3 years,5 years were 75. 4%,47. 3%, 32. 6% in RFA+TACE group,and 63. 1%,37. 2%,22. 4% in RFA group. Conclusion For patients with early hepatocellular carcinoma with liver cirrhosis,RFA combined with TACE have higher tumor free survival rate than RFA alone in the treatment of early hepatocellular carcinoma,but no difference in overall survival rate. The results still need to be validated by prospectively randomized controlled trials.

Objective To investigate the management of pseudomyxoma peritonei originated in the appendix.Methods The clinical data of 51 patients with pseudomyxoma peritonei originated in the appendix who were admitted to the Beijing Hospital from 1970 to 2010 were retrospectively analyzed.The results of operation,reoperation,adjuvant treatment and follow-up were analyzed.The time from pseudomyxoma peritonei recurrence to the reoperation between patients who did or did not receive chemotherapy was compared by two tailed t test.Results Of the 51 patients,48 received operation,and the operation time was (135 ± 72 )minutes.Tumor recurrence was observed in 34 patients,and 16 of them received cytoreduction procedure,and 33 cytoreduction procedures were performed in total.The median time of follow-up was 49.7 months (range,3-132 months).The disease-specific survival was observed in 25 patients and disease-free survival in 16 patients.Four patients died of tumor recurrence or progression.The results of postoperative pathological examination confirmed that 19 patients were with benign disseminated peritoneal adenomucinosis (DPAM),26 were with malignant peritoneal mucinous carcinomatosis (PMCA) and 6 were with intermediate subtype (PMCA-1).The 3-,5- and 10-year survival rates were 75％ (38/51),55％ (28/51) and 22％ ( 11/51 ),respectively.The survival time and reoperation time interval for patients who received postoperative chemotherapy were ( 21 ± 4) months and ( 10 ± 6 ) months,which were longer than (19 ±7 )months and (7 ±4)months of those who did not receive postoperative chemotherapy (t =1.027,0.361,P ＞ 0.05).The median survival time of patients with benign DPAM,PMCA-1 and malignant PMCA were 96,63,23 months,respectively.The tumor recurrence interval for patients with benign DPAM and those with malignant PMCA were ( 15 ± 5 ) months and (7 ± 4) months,with significant difference ( t =2.193,P ＜ 0.05 ).Conclusions An active cytoreduction surgery is feasible for patients with pseudomyxoma peritonei originated in the appendix in improving survival.Repeated cytoreduction is a treatment of strategy to prolong the recurrence time and improve the prognosis of selected patients.

AIM: To investigate the effect of glucocorticoid receptor beta(GR?) on the transcriptional activation function of glucocorticoid receptor alpha(GR?) and elucidate the biological significance of glucocorticoid receptor beta. METHODS: GR? (pRSH-GR?) and GR? responsive reporter gene cat (pMMTV-cat) were cotransfected into human osteosarcoma cell line (HOS-8603). Transient transfectants were treated with dexamethasone and the activity of cat was determined by the method of ELISA. The expression p21 messenger RNA was detected by the method of reverse transcription-polymerase chain reaction. RESULTS: GR? repressed the expression of reporter gene-cat in a dose-dependent manner. The expression of messenger RNA of p21, an endogenous target gene of GR?, was also inhibited by GR?. CONCLUSION: GR? can inhibit the transcriptional activation function of GR?. GR? may be an endogenous inhibitor of GR?.

AIM: To explore the possible role of p21 wafl gene in the effect of 1,25-dihydroxyvitamin D 3 on a human osteosarcoma cell line (HOS-8603) proliferation and differentiation. METHODS: The effect of 1,25(OH) 2D 3 on the expression of p21 wafl mRNA was determined by quantitative reverse transcription-polymerase chain reaction. The human osteosarcoma cell line stably expressing the p21 wafl mRNA, which named HOS-p21 wafl , was constructed by lipofectamine transfection, and the expression of p21 wafl protein was detected by Western blot. Then the growth curve of the HOS-p21 wafl cells and the expression of alkaline phosphatase (AKP) in the HOS-p21 wafl cells were investigated. RESULTS: The expression of endogenous p21 wafl mRNA in HOS-8603 cells was induced by 1,25(OH) 2D 3 in a time-dependent manner, reaching the maximum at 4 h, and remaining the inducible action for up to 24 h over basal levels. The HOS-p21 wafl cells grew much slower than the control cells, only reaching 50% of the control ones when cultured for up to 6 days. Histochemical analysis showed that the expression of alkaline phosphatase (AKP) in the HOS-p21 wafl cells increased remarkably.CONCLUSION: These results suggest that the p21 wafl mRNA expression induced by 1,25(OH) 2D 3 is one of important mechanisms responsible for the cellular effects of the hormone on HOS-8603 cells.

Objective To investigate the effects of exosomes from cultured human retinal pigment epithelium (ARPE-19) cells affected by oxidative stress on the proliferation and expression of vascular endothelial growth factor-A (VEGF-A) and Akt of ARPE-19 cells. Methods Culture ARPE-19 cells. The concentration of 2.5μmol/L rotenone was selected to simulate oxidative stress and isolated ARPE-19-exosome. Exosomes were isolated by ExoQuick exosome precipitation solution. Transmission electron microscopy was used to identify the morphology of exosomes. Western blot was used to detect exosomes’ surface-specific maker protein CD63. ARPE-19 cells affected by oxidative stress were cultured with exosome as experimental group, normal ARPE-19 cells were cultured with exosome as control group. The cell proliferation was examined by methyl thiazolyl tetrazolium assay. Western blot and immunofluorescence assay were used to detect the expression levels of VEGF-A and Akt protein. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the levels of VEGF-A mRNA and Akt mRNA. Results The diameter of normal ARPE-19-exosomes ranged from 50 to 150 nm. The isolated exosomes expressed CD63. AREP-19 cells were cultured with ARPE-19 (affected by rotenone)-exosome, the cell viability in experimental group was significantly reduced than in the control group. Green fluorescence was observed in the cytoplasm under fluorescence microscope. Compared with the control group, VEGF-A was up-regulated expressed and Akt was down-regulated expressed. Western blot results showed that, VEGF-A protein expression in the experimental group were higher than the control group. Akt protein expression in the experimental group were less than the control group. The difference was statically significant (t=3.822, 6.527;P<0.05). RT-PCR results showed that VEGF-A mRNA expression levels was higher in the experimental group than the control group. Akt mRNA expression levels was lower in the experimental group than the control group. The difference was statically significant (t=8.805,?7.823;P<0.05). Conclusions Exosomes from ARPE-19 cells affected by oxidative stress inhibit the proliferation of normal ARPE-19 cells, increase the expression of VEGF-A and reduce the expression of Akt.

Objective To investigate the effect of arginase (Arg) inhibitor N-ω-Hydroxy-L norArginine (nor-NOHA) on high glucose cultured rhesus macaque retinal vascular endothelial cell line (RF/6A) in vitro.Methods The RF/6A cells were divided into the following 4 groups:normal control group (5.0 mmol/L of glucose,group A),high glucose group (25.0 mmol/L,group B),high glucose with 125 mg/L nor-NOHA group (group C),and high glucose with 1％ DMSO group (group D).The proliferation,migration ability and angiogenic ability of RF/6A cells were measured by Methyl thiazolyl tetrazolium (MTT),transwell chamber and tube assay respectively.The express of Arg Ⅰ,eNOS,iNOS mRNA of RF/6A cells were measured by real-time polymerase chain reaction (RT-PCR),Enzyme-linked immuno sorbent assay (ELISA) was used to detect the expression of NO and interleukine (IL)-1b of RF/6A cells.Results The proliferation,migration,and tube formation ability of group A (t=2.367,5.633,7.045;P＜0.05) and group C (t=5.260,6.952,8.875;P＜0.05)were significantly higher than group B.RT-PCR results showed the Arg Ⅰ and iNOS expression in group B was higher than that in group A (t=6.836,3.342;P＜0.05) and group C (t=4.904,7.192;P＜0.05).The eNOS expression in group B was lower than that in group A and group C (t=4.165,6.594;P＜0.05).ELISA results showed NO expression in group B was lower than that in group A and group C (t=4.925,5.368;P＜0.05).IL-1b expression in group B was higher than that in group A and group C (t=5.032,7.792;P＜0.05).Conclusions Nor-NOHA has a protective effect on cultured RF/6A cells in vitro and can enhance its proliferation,migration and tube formation.The mechanism may be inhibiting the oxidative stress by balancing the expression of Arg/NOS.

Objective To seek the predictive value of pudendal nerve function that need preventive anti-incontinence surgery at the same time following pelvic prolapse surgery in severe pelvic organ prolapse (POP) patients. Methods Seventy women completed this study from January 2014 to June 2015 in Fuzhou General Hospital of Nanjing Military Command, dividing into four groups: POP with or without coexisting occult stress urinary incontinence (OSUI) in preoperation, women with persistent stress urinary incontinence (SUI) in postoperation, women without SUI in postoperation. The pudendal nerve function in preoperation was measured by using Solar Urodynamic Neuro Module, including pudendal nerve terminal motor latency (PNTML), and amplitude. Results There were statistical significance on bilateral PNTML between POP coexisting OSUI group and only severe POP group [(2.62±0.23) versus (2.40±0.26) ms in right of PNTML, (2.55 ± 0.21) versus (2.37 ± 0.30) ms in left of PNTML; all P<0.05], but no statistical significance on bilateral amplitude (P>0.05). Compared de novo SUI group with POP group in postoperation, de novo SUI group′s right of PNTML was significantly increased [(2.74 ± 0.16) versus (2.47 ± 0.26) ms; P<0.05]; and the right of PNTML was extending 2.5 standard deviation at least compared with the health′s [(2.10±0.20) ms]. Conclusions The PNTML of pudendal nerve of POP coexisting OSUI is severe than only severe POP, the velocity of nerve conduction is slowing, and PNTML extension has a predictive value for postoperative urinary incontinence. When the right of PNTML of preoperative POP increased by at least 2.5 standard deviations than health′s, the risk of SUI postoperative strongly increased, and a anti-incontinence surgery at the same time following pelvic prolapse surgery should be adviced.