Objective To observe enhancing effect of nerve regeneration on peripheral nerve defect models bridged by a new PRGD/PDLLA/VPA composite conduit.Methods In this study from February,2012 to March,2014,PRGD/PDLLA/VPA nerve conduits were tested in the rat sciatic nerve transection model.At different periods after operation,its ability to promote nerve regeneration was evaluated by sciatic functional index(SFI),electrophysiology (CMAPs,NCVs) and histologic assessment.Forty rats were randomly divided into 4 groups (n =10),group A:PRGD/PDLLA/VPA,group B:PDLLA/VPA,group C:PRGD/VPA and group D:autograft.Results At 12 weeks after surgery,the SFI value of group A (-45 ± 3.19)and group D (-42 ± 3.01)were significantly higher than those of group B(-79 ± 3.06) and group C(-72 ± 2.07)(P ＜ 0.05);The CMAPs of group A (24.89 ± 5.01) and group D (25.39 ± 5.63) were significantly higher than those of group B(14.88 ± 3.11) and C(15.00 ± 5.54);the NCVs of group A (31.42 ± 2.43) and group D (31.50 ± 2.16) were significantly higher than those of group B (20.11 ± 2.39) and group C(21.00 ± 2.13)(P ＜ 0.05).At 12 weeks after surgery,the numbers of regenerated nerve in the tube of group A (258 ± 6.18) and D(259 ± 5.59) were significantly higher than those of group B (231 ± 5.00) and group C(230 ± 5.07)(P ＜ 0.05).There was no significant difference between groups A and D(P ＞ 0.05).Conclusion These results illustrated that this new PRGD/PDLLA/VPA conduit could significantly facilitate the regeneration of short nerve defect and recovery of motor nerve,which provides a new thought for treatment of peripheral nerve injury.

Objective To explore the expression of PIWIL1 protein and DICER enzyme in hepa tocellular carcinoma (HCC) and their significance.Methods Immunohistochemical method was used to detect the expression of PIWIL1 and DICER in 47 cases of HCC and the adjacent HCC tissues.Western blot method was used to detect the expression of PIWIL1 and DICER in 31 cases of fresh HCC tissues and their adjacent HCC tissues.The relationship between PIWIL1 and DICER and their relationships were analysed with clinical features.12 cases of normal liver tissues were used as control group.Results The expression of PIWIL1 was high in HCC but low in normal liver tissues (P＜ 0.05).The expression of DICER was high in normal liver tissues but low in HCC (P＜0.05).The expression of PIWIL1 was positively correlated with invasion to adjacent tissues and histological differentiation (P＜0.05).The expression of DICER was negatively correlated with invasion to the adjacent tissues and histological differentiation (P＜0.05).There was a negative correlation between PIWIL1 and DICER (P＜ 0.05).Conclusions High expression of PIWIL1 and low/missing expression of DICER was related to pathological differentiation and invasion of adjacent tissues.

AIM: To investigate the clinical effect for treating bullous keratopathy ( BK ) by anterior corneal stromal acupuncture combined with amniotic membrane transplantation.
METHODS: Totally 35 patients ( 35 eyes ) with bullous keratopathy were treated by corneal stromal acupuncture combined with amniotic membrane transplantation. All patients preoperative and postoperative underwent anterior segment OCT, corneal topography, corneal sensitivity and confocal microscopy. To observe postoperative ocular symptoms in patients with BK, recurrence of bulla, changes in corneal thickness, corneal sensitivity and changes in the organizational structure of the layers of the cornea.
RESULTS: Thirty-five were followed up for 6-18mo. The symptom of pain disappeared in 32 cases ( 91%) in the first day after operation and did not recur during follow-up. The symptom of pain relieved in 3 cases (9%) in the first day after operation and disappeared in 3 days. Corneal epithelium of 9 cases ( 26%) were all healed within 1wk, 21 cases (60%) were all healed within 2wk, and 5 cases (14%) were all healed within 3wk. Following up for 6-18mo, there was no recurrence of symptoms or bulla. A small amount of tiny bubbles in the surrounding area appeared in 2 cases after 3 and 4wk. All patients had no neovascularization, and had smooth corneal surface. The amnions of 30 cases (86%) were thinning after 2mo, partially dissolved and absorped, not seen with the naked eye after 3mo. Thirty-four cases (97%) had no changes in vision, one case (3%) was from the light to front of the manual. After 2mo, corneal sensation decreased in 30 cases ( 86%) , corneal thickness increased from preoperative 788±35μm to 940±43μm. After 12mo, corneal thickness increased to 1060±27μm. Results of confocal microscopy: after 3mo, the number density of the trigeminal nerve fibers under corneal basement membrane reduced, shallow stromal cells became into fibrotic stroma, deep stroma was more loose, and cells swelled significantly. The number of endothelial cells reduced and form swelled more obviously compared with preoperatively.
CONCLUSION: Corneal acupuncture combined with amniotic membrane transplantation can effectively control the symptoms of BK, prevent the recurrence of BK, and especially it is a simple, safe and practical way for patients with poor visual function.

Objective To investigate the diagnostic value of urinary light chain detection in essential hypertension renal injury. Methods According to "National Prevention and Treatment Guide of Hypertension" (2005 Edition) and the diagnostic criteria of essential hypertension renal injury, patients'medical history and damage of target organs, we selected 60 patients with essential hypertension and divided them into two groups: 28 patients without renal injury and 32 patients with renal injury. Meanwhile, Thirty healthy individuals were included in a control group. Urinary proteins and light chains were detected by immuonephelometry and the activity of NAG was determined by liquid kinetic rate assay with MNP as substrate. The urinary light chains concentration of each group were compared and analyzed with those of Alb and α1-MG. Results The results of lamU in renal injury group [8. 89(3.84-33. 30) mg] were higher than those of non renal injury group [3. 84(3. 84-10. 4) mg] and control group [3.84(0. 00-3.84) mg]. There was statistically significant difference ( H = 37.97, P ＜ 0. 01 ). The levels of kapU in renal injury group [26. 65 (6. 97-62. 5 ) mg] were higher than those of non renal injury group [6. 97 (6. 97-23.7 )mg] and control group [6. 97(6. 97-7. 20) mg], which also had statistical significance (H =40. 09,P ＜0. 01 ). All the other results in renal injury group ( Alb, α1-MG, IgG, TRF and NAG) were higher than those of non renal injury group and control group ( H =21.02, 32. 65, 25.50, 30. 45 and 30. 57, P ＜0. 01 ). Correlation analysis showed that Alb levels were highly correlated with urinary IgG and TRF levels [Correlation coefficient (r) =0. 911, 0. 965, P ＜ 0. 01]. The levels of urinary light chain lamU and kapU were also highly correlated with α1-MG ( r = 0. 804, 0. 827, P ＜ 0. 01 ). However, There was no correlationship between NAG and α1MG levels (r=0. 398,P＞0. 05). Conclusion The urinary light chain lamU and kapU can be used as indicators for evaluation of renal injury in essential hypertension, especially renal tubular injury.

Objective　To ascertain the natural infection rate of hemorrhagic fever with renal syndrome virus (HFRSV) among its hosts and the type of the natural foci for providing some baseline data for the immigrant health and epidemic prevention of the Three-Gorge region. Methods　Epidemiological survey on the field was performed including epidemiological data collection, ecology of rodents and pathogen detection. HFRS virus antigen of hosts were detected by the direct immunofluorescent assay (DIFA) technique and determination of HFRSV-RNA by ISH were carried out from HFRSV-Ag-positive animals. Results　HFRSV-Ag-positive animals were found in 5 migration areas ie Baitao Town of Fuling Section, Wansheng Village of Fengjie County and Dachang Town of Wushan County. The positive hosts were as follows, Rattus Norvegicus, Apodemus agrarius, Anourusurex squamipes, Mus musculus and Rattus flavipectus. The positive rate of HFRSV in the mice of 5 migration areas were 19.4%, 17.0%, 14.0%, 13.7%, and 8.5% respectively. The results showed that the lung tissues of some hosts in all five migration areas were HFRSV-RNA-positive. Baitao Town and Peishi Town were attributed to mixture type epidemic areas while. Kangle Town, Wansheng Village and Dachang Town were domestic rats type epidemic areas. Conclusion　This study shows that the five migration areas are natural epidemic foci of HFRS. It is predicted that maximum risk of HFRS breakout or epidemic may take place after the completion of the San Xia Reservoir(the Three-Gorges Reservoir), which results from rodent moving toward higher land. Therefore, deratization and preventive measures for rat are important in migration areas.

OBJECTIVETo study the regularity of migration and distribution of bone marrow stromal cells (BMSCs) in injured spinal cord with intradural space transplantation.

METHODSForty Wistar rats were randomly assigned into 4 groups. The spinal cord injury model was prepared according to the modified Allen method. BMSCs were labeled by CM-Dil. And 5.0 multiply 10(6) cells were transplanted by different channels including intraventricular injection (Group A),injured spinal cord intrathecally injection (Group B), remote intrathecally injection at the L(3)-L(4) level (Group C), and intravenous injection (Group D). Spinal cord was dissected at 24 hours, 1, 2, 3 and 4 weeks after transplantation. Sections of 4 micromolar were cut on a cryostat and observed under fluorescence microscopy.

RESULTSNo fluorescence was observed 24 hours after transplantation in spinal cord injury parenchyma except Group B. One week later, BMSCs in Groups A and C began to migrate to the injured parenchyma; 2-4 weeks later, BMSCs penetrated into the injured parenchyma except Group D. The number of BMSCs decreased at 3-4 weeks after transplantation. The number of cells in Group B decreased faster than that of Groups A and C.

CONCLUSIONSBMSCs transplanted through intraventricular injection, injured spinal cord intrathecally injection and remote intrathecal injection could migrate to the injured parenchyma of spinal cord effectively. The number of BMSCs migrated into injured spinal cord parenchyma is rare by intravenous injection.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; methods ; Cell Movement ; physiology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; pathology ; surgery ; Stromal Cells ; cytology ; transplantation

OBJECTIVETo investigate the impact of experimental varicocele (EV) on the ipsilateral testis in rats.

METHODSEV was induced by partial ligation of the left renal vein in male SD rats, the control rats subjected to sham operation, and the testes of the EV models and controls were extirpated 6, 12, and 18 weeks later. Johnson's score, ultrastructure of seminiferous tubules, intratesticular testosterone concentration (ITC) and germ cell apoptotic index (AI) of each left testis were evaluated.

RESULTSJohnson's scores were (6.92 +/- 0.52), (4.83 +/- 0.41) and (2.95 +/- 0.26), ITCswere (6.32 +/- 0.85), (5.17 +/- 0.76) and (4.11 +/- 0.69) and AIs were (5.32 +/- 1.23), (15.21 +/- 0.97) and (21.13 +/- 1.12) respectively in the 6 w , 12 w and 18 w EV groups, significantly lower than in the corresponding control groups, (9.56 +/- 0.35, 9.63 +/- 0.31, 9.39 +/- 0.46), (9.64 +/- 1.23, 9.38 +/- 0.69, 9.73 +/- 0.49) and (3.21 +/- 1.15, 3.43 +/- 1.21, 3.61 +/- 1. 15) (P < 0.05), the former two showing a gradual decline while the latter a significant elevation with the increasing duration of varicocele. The damage to the ultrastructure of seminiferous tubules was aggravated with the prolonging of varicocele.

CONCLUSIONEV can cause a progressive decline of ITC, dyszoospermia and increased AI of germ cells.

Animals ; Apoptosis ; Disease Models, Animal ; Infertility, Male ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Epithelium ; cytology ; ultrastructure ; Testis ; chemistry ; metabolism ; Testosterone ; metabolism ; Varicocele ; metabolism ; physiopathology

OBJECTIVETo project the future impacts of climate change on heat-related mortality in shanghai.

METHODSThe statistical downscaling techniques were applied to simulate the daily mean temperatures of Shanghai in the middle and farther future under the changing climate. Based on the published exposure-reaction relationship of temperature and mortality in Shanghai, we projected the heat-related mortality in the middle and farther future under the circumstance of high speed increase of carbon e mission (A2) and low speed increase of carbon emission (B2). The data of 1961 to 1990 was used to establish the model, and the data of 1991 - 2001 was used to testify the model, and then the daily mean temperature from 2030 to 2059 and from 2070 to 2099 were simulated and the heat-related mortality was projected. The data resources were from U.S. National Climatic Data Center (NCDC), U.S. National Centers for Environmental Prediction Reanalysis Data in SDSM Website and UK Hadley Centre Coupled Model Data in SDSM Website.

RESULTSThe explained variance and the standard error of the established model was separately 98.1% and 1.24°C. The R(2) value of the simulated trend line equaled to 0.978 in Shanghai, as testified by the model. Therefore, the temperature prediction model simulated daily mean temperatures well. Under A2 scenario, the daily mean temperature in 2030 - 2059 and 2070 - 2099 were projected to be 17.9°C and 20.4°C, respectively, increasing by 1.1°C and 3.6°C when compared to baseline period (16.8°C). Under B2 scenario, the daily mean temperature in 2030 - 2059 and 2070 - 2099 were projected to be 17.8°C and 19.1°C, respectively, increasing by 1.0°C and 2.3°C when compared to baseline period (16.8°C). Under A2 scenario, annual average heat-related mortality were projected to be 516 cases and 1191 cases in 2030 - 2059 and 2070 - 2099, respectively, increasing 53.6% and 254.5% when compared with baseline period (336 cases). Under B2 scenario, annual average heat-related mortality were projected to be 498 cases and 832 cases in 2030 - 2059 and 2070 - 2099, respectively, increasing 48.2% and 147.6% when compared with baseline period (336 cases).

CONCLUSIONUnder the changing climate, heat-related mortality is projected to increase in the future;and the increase will be more obvious in year 2070 - 2099 than in year 2030 - 2059.

China ; Climate Change ; Greenhouse Effect ; Humans ; Models, Theoretical ; Mortality ; Risk Assessment

OBJECTIVE Gastric cancer is one of the most common malignant tumors,and the inci-dence rate is the highest in all kinds of tumors in China. However,it remains unclear that how signifi-cantly gastric cells are dependent on glycolysis,and which type of gastric cells are sensitive to glycolysis inhibition. In this study, several kind of gastric cancer cell lines were used as the research object, and the metabolic characteristics of different cell lines were systematically analyzed to provide theoretical support for the accurate treatment of gastric cancer. METHODS We examined the energy metabolism of four gastric cancer cell lines(MGC-803,SGC-7901,HGC-27 and BGC-823)by using glycolysis inhibitor, 2-deoxy-D-glucose(2-DG)and inhibitor of oxidative phosphorylation,oligomycin.Oxygen consumption rates(OCR)and L-lactate were also measured with an XF96 Analyzer(Seahorse Biosciences)to deter-mine the significance of metabolism of oxidative phosphorylation and aerobic glycolysisin gastric cells. In addition, western blot was used to detect the contribution of AMP-activated protein kinase (AMPK), and anti-apoptotic proteins(Bcl-2 and survivin)to clarify the mechanism of death or survival of gastric cancer cells treated by 2-DG or oligomycin. RESULTS In this study, it was shown that the growth of gastric cell lines were suppressed by 2-DG.However,the sensitivity to 2-DG was quite different among cell lines:IC 50 of 2-DG was from 3.28 mmol·L-1(MGC-803)to 15.57 mmol·L-1(BGC-823).MGC-803 was relatively sensitive to 2-DG (IC 50:3.28 mmol·L-1), consumed more glucose and produced more lactate (waste product of glycolysis) than the three other cell lines. Consequently, MGC-803 could be more dependent on glycolysis than other cell lines, which was further confirmed by the fact that glucose (+)FCS(-)medium showed more growth and survival than glucose(-)FCS(+)medium.Alternatively, BGC-823, most resistant to 2-DG (IC50: 15.57 mmol·L- 1), was most sensitive to oligomycin, and showed more growth and survival in glucose(-)FCS(+)medium than in glucose(+)FCS(-)medium. Thus,we had reasons to think BGC-823 cells depended on oxidative phosphorylation for energy production. In BGC-823,AMPK,which is activated when ATP becomes limiting,was rapidly phosphorylated by 2-DG, and expression of Bcl-2 was augmented,which might result in resistance to 2-DG.Interestingly,AMPK phosphorylation and augmentation of Bcl-2 expression by 2-DG were not observed in MGC-803,which is 2-DG sensitive. CONCLUSION There is a large metabolic difference between gastric cancer cell lines,which will facilitate the future gastric cancer therapy by targeting metabolic pathways.

OBJECTIVE To probe into the anti-esophagus cancer activity and mechanisms of DN3, a novel natural diterpenoid derivative. METHODS The anti-tumor activity in vitro of DN3 was evaluated by MTT, and by using human esophageal carcinoma cells xenografted into athymic mice model in vivo. The specific mechanisms of DN3, as a dual inhibitor of glycolysis and oxidative phos-phorylation(OXPHOS)were explored through cell and molecular biology techniques.For instance,the manner of cancer cell death induced by DN3 was characterized by hoechst33342, FITC-Annexin V/PI staining and flow cytometric analysis,then these changes of glucose consumption,glucose uptake and lactate production in glycolysis, as well as oxygen consumption rate (OCR) and ATP content in OXPHOS caused by DN3 were performed separately through related kits and SeahorseBioscience XF24 Extra-cellular Flux Analyzer.Furthermore,in order to obtain a clear understanding of the inhibition of DN3 to glycolysis and OXPHOS, these regulatory factors were investigated by Western blot, such as PI3K/AKT, c-Myc and p53 of glycolysis, Bax and HK2 of mitochondrial function. RESULTS DN3 inhibited the growth of esophagus cancer cell EC9706, EC109 and EC1 cells in a dose and time dependent manner,but showed no significant effects on human esophageal epithelial cells(HEECs).DN3 caused significant G2/M arrest of esophagus cancer cell lines and induced apoptosis of these cell lines, which indicated DN3 inhibited the growth of esophagus cancer cell through blocking cell cycle and inducing apoptosis in a dose and time-dependent manner. Importantly, 8 μM DN3 decreased the extracellular acidification rate (ECAR) by 45% in EC109, which indicated glycolysis was inhibited by DN3. Mean-while, DN3 decreased the oxygen consumption rate (OCR) and the OCR linked to intracellular ATP production in EC109 cells,but that was not obvious in HEECs,so which indicated that DN3 could selec-tively block OXPHOS of cancer cells. In addition, the accumulation of reactive oxygen species (ROS) and the drop of mitochondrial membrane potential (MMP) were also observed in EC109 incubated by DN3,which suggested mitochondrial biological function was disturbed.Furthermore,the expression of PI3K/AKT, c-Myc and HK2 related to glycolysis were down-regulated by DN3, but the p53 and Bax were up-regulated in esophageal carcinoma cells. The changes of these enzymes accounted for the decreased glycolysisand OXPHOS in esophageal carcinoma cells treated by DN3. CONCLUSION The new compound DN3 has a strong anti-esophageal carcinoma activity,and it is tolerable that DN3 is seen as a dual inhibitor of glycolysis and oxidative phosphorylation.