Objective To explore the protective effect in a model of nicotinamide riboside(NR)against carbonyl cyanide m-chlorophenylhydrazone(CCCP)-induced oxidative stress in R28 cells.Methods 4 μmol/L CCCP was used to induce oxidative stress in R28 cells,and 400 nmol/L NR was used to intervene.The cell viability was quantified by CCK-8 assay.The apoptosis was detected by Annexin-V/PI double staining and flow cytometry.Western blotting was used to examine the levels of Cytochrome C,Caspase-3,and Caspase-9 to evaluate the apoptosis.Tetramethylrhodamine ethyl ester was used to detect the mitochondrial membrane potential(MMP),MitoSOX was used to detect the mitochondrial reactive oxygen species(mtROS)levels,and adenosine triphosphate(ATP)assay kit was used to assess ATP generation ability to evaluate mitochondrial function.Results After CCCP treatment of R28 cells,the cell viability decreased,the apoptotic protein levels and apoptosis rates increased,the MMP decreased,and the mtROS generation increased(P＜0.05).After NR pretreatment,the cell viability increased,the apoptotic protein levels and apoptosis rates decreased,the MMP increased,and the mtROS generation decreased(P＜0.05).Conclusion:NR enhances the cell viability,reduces the expression of apoptotic proteins,and ultimately reduces the apoptosis of retinal ganglion cell by inhibiting oxidative stress response and protecting mitochondrial function.

Objective:To predict and analyze the T/B combined epitope of EM10 protein in Echinococcus multilocularis, and identify the expressed products of the biosynthetic EM10 multi epitopes. Methods:The gene-related information of EM10 protein was obtained through NCBI GenBank public database. Bioinformatics technique was used to predict and analyze the T/B binding epitopes of EM10 protein. The prokaryotic expession recombinant plasmid pET30a-EM10 (epitope) was synthesized, and transformed into host bacteria Ecoli. BL21 (DE3). The expression of EM10 recombinant multi-epitope protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting after induced expression by isopropyl thiogalactopyranoside (IPTG). Results:The total length of EM10 gene was 1 759 bp (GenBank registration number: U05573), and its protein amino acid sequence (GenBank registration number: AAA50580.1) was 559 amino acids. By using Phyre software for homology modeling, the tertiary structure of EM10 protein was obtained, and the T/B combined epitope of EM10 protein was successfully predicted, the dominant epitope was located at 46 - 61, 133 - 183, 239 - 255 and 442 - 475 amino acid sites. The (GGGGS)n linker sequence was used to connect the epitopes to form an EM10 recombinant multi-epitope protein with a total of 206 amino acid. The size of the DNA fragment was 618 bp and the relative molecular weight of the protein was 22.66 × 10 3. The prokaryotic expession recombinant plasmid was validated by enzyme digestion, the results showed that the plasmid size was between 5 000 and 6 000 bp, which was consistent with the length of the constructed plasmid (5 854 bp). SDS-PAGE showed that the target protein was expressed in the supernatant induced by IPTG at 37 ℃ and the effect was the best. The relative molecular weight of the protein was 22.66 × 10 3 by Western blotting, which was consistent with the constructed plasmid. Conclusions:The combined epitope of EM10 T/B is successfully designed and predicted using bioinformatics technology. A prokaryotic expression recombinant plasmid is constructed, the expression of EM10 recombinant multi-epitope protein is verified through experiments, providing an experimental basis for the construction of an EM10 dominant epitope diagnostic kit.

Zinc levels are high in pancreatic β-cells, and zinc is involved in the synthesis, processing and secretion of insulin in these cells. However, precisely how cellular zinc homeostasis is regulated in pancreatic β-cells is poorly understood. By screening the expression of 14 Slc39a metal importer family member genes, we found that the zinc transporter Slc39a5 is significantly down-regulated in pancreatic β-cells in diabetic db/db mice, obese ob/ob mice and high-fat diet-fed mice. Moreover, β-cell-specific Slc39a5 knockout mice have impaired insulin secretion. In addition, Slc39a5-deficient pancreatic islets have reduced glucose tolerance accompanied by reduced expression of Pgc-1α and its downstream target gene Glut2. The down-regulation of Glut2 in Slc39a5-deficient islets was rescued using agonists of Sirt1, Pgc-1α and Ppar-γ. At the mechanistic level, we found that Slc39a5-mediated zinc influx induces Glut2 expression via Sirt1-mediated Pgc-1α activation. These findings suggest that Slc39a5 may serve as a possible therapeutic target for diabetes-related conditions.

[Objective]To analyze the prognostic factors of resectable acral melanoma patients ,then develop a novel prognostic model and examined its prognostic value.[Methods]The study retrospectively analyzed clinicopathological characteristics and inflam?matory markers of 232 acral melanoma patients who underwent radical surgical resection between 2000 and 2011 at the Sun Yat-sen University Cancer Center. Kaplan-Meier curves were plotted to estimate overall survival. Significantly predictive factors were identified by multivariate Cox regression analyses and a prognostic model based on these variables was constructed to predict survival.[Results]Cox regression analysis revealed that age,lactic dehydrogenase(LDH),stage,globulin(GLB)and C-reactive protein (CRP)were independently related to survival. After computing these scores ,patients were classified into three risk groups. The new prognostic model identified three categories of patients with different prognoses(P<0.001)and significantly stratify patient prognosis in different tumor stages. The 5-year survival rate was 42.9%,25.7%,and 3.7%in groups 1,2,and 3,respectively. The AUC of new prognostic model is 0.664(95%CI:0.599-0.724).[Conclusion]Age,LDH,stage,GLB and CRP were independently related to survival in our study population,and the prognostic model is useful to stratify patients into different risk groups and it is a useful complement to AJCC staging for Asian patients with acral melanoma.

Objective To evaluate the therapeutic effect of oral and external application of Chinese medicine combined with Ilizarov external fixator and the accordion technique for severe chronic osteomyelitis of the tibia.Methods A total of 78 patients with severe chronic osteomyelitis of the tibia were randomized into a routine treatment group and a combined treatment group, 39 in each group. All the patients in the two groups received external fixation by use of Ilizarov external fixator and the accordion technique. All the patients in the combined treatment group received oralGuyu decoction. The patients who had large wound received vacuum-sealing drainage in the routine treatment group, and vacuum-sealing drainage combined with external application ofShengji-Yuhong plaster in the combined treatment group. All the patients were treated for 8 weeks and followed up for 2 years. The time to wound healing and fracture healing, and the drainage time were compared between the two groups. Functional and radiologic findings were evaluated according to Paley's criteria. The functions of the knee joint and ankle joint were evaluated using the Hospital for Special Surgery (HSS) knee score and the scoring system of Baird and Jackson, respectively.Results The time to wound healing (17.33 ± 6.21 dvs. 22.27 ± 8.12 d;t=3.018,P=0.004) and the time to fracture healing (32.25 ± 6.02 weeks vs. 36.37 ± 7.75 weeks;t=2.623,P=0.011), and the drainage time (17.01 ± 4.66 dvs. 21.51 ± 5.23 d;t=4.012, P<0.001) in the combined treatment group were significantly shorter than those in the routine treatment group. According to Paley's criteria, the patients who achieved a score of excellent or good for fracture healing (84.6%vs. 53.8%;χ2=7.282,P=0.007) and function (92.3%vs. 66.7%;χ2=6.369,P=0.012) in the combined treatment group were significantly more than those in the routine treatment group. The scores of the HSS knee score (84.56 ± 7.42vs. 78.81 ± 5.33;t=3.391,P=0.002) and the scoring system of Baird and Jackson (85.01 ± 8.21vs. 79.21 ± 6.78;t=3.402,P=0.024) in the combined treatment group were significantly higher than those in the routine treatment group.Conclusion OralGuyu decoction and external application ofShengji-Yuhong plaster combined with Ilizarov external fixator and the accordion technique can promote recovery of the joint function, fracture healing and wound healing.

Heterotopic ossification is a common complication after acetabular fractures and fractures of the elbow.Heterotopic ossification often leads to severe joint movement disorder,which brings great pain to the patient.This paper reviewed the clinical research,including pathogenesis,clinical diagnosis,prevention,treatment and future directions of heterotopic ossification to investigate the effective method in prevention and treatment of heterotopic ossification.

This paper briefly introduced seven cases of malfunctions occurred in the VARIAN IX linear accelerator during operation, i.e., control board malfunction of electronic gun (GFIL interlock), modulator malfunction (HVCB interlock), energy programming board malfunction (EXQ1), energy conversion potentiometer malfunction (CARR interlock), MLC malfunction, thermostat valve malfunction (PUMP interlock) and ionization chamber malfunction (ION interlock). Moreover, malfunction cases analyzed and troubleshooting methods presented in this paper can provide reference for coleagues.

Objective To detect the serum expression level of Pokemon and explore its clinical value combines with AFP,AFP-L3 or DCP in hepatocellular carcinoma (HCC).Methods In this study,the serum levels of Pokemon,AFP,AFP-L3 and DCP in 30 patients with HCC,34 patients with benign liver diseases and 30 healthy controls were examined by enzyme-linked immunosorbent assay.Then,the clinical significance of Pokemon expression level was analyzed.In the end,the diagnostic evaluation of four serum biomarker alone detection was used to analyze the sensitivity,specificity,the area under the curve and Youden index in the diagnosis of HCC.Results The serum expression level of Pokemon in patients with HCC was (22.63 ± 6.82) ng/ml,which was significantly higher than that in the group with benign liver diseases and healthy controls [(3.54±1.26)ng/ml,t =8.594,P＜0.001;(1.95 ±0.57)ng/ml,t =10.021,P＜0.001].The expression level of Pokemon in HCC patients was correlated to tumor size (t =2.678,P =0.021) and TNM stag (t =2.578,P =0.034).The serum expression levels of AFP,AFP-L3 and DCP in patients with HCC were (774.56 ± 96.52) ng/ml,(26.37 ± 2.54) ng/ml and (93.49 ± 30.45) mAU/ml,which were significantly higher than those in the group with benign liver diseases [(22.21 ± 3.57) ng/ml,(7.05 ± 2.71) ng/ml,(34.68 ± 10.13) mAU/ml] and healthy controls [(14.65 ± 4.45) ng/ml,(3.84 ± 1.09) ng/ml,(25.87 ± 9.01)mAU/ml,F=58.46,P=0.000;F=12.47,P=0.000;F=23.41,P=0.000].The sensitivity and specificity of Pokemon,AFP,AFP-L3,DCP in the diagnosis of HCC were 84.6％ and 87.5％,52.5％ and 78.9％,95.0％ and 80.0％,77.1％ and 87.5％,respectively.Conclusion The enhanced Pokemon serum level,with its sensitivity and specificity higher than AFP,points out Pokemon may be a potentially useful biomarker to diagnose HCC.

Objective To study the dosimetry characteristics of VMAT plan in the esophageal carcinoma radiotherapy.Methods Application of 0.6 cm3 ionization chamber and COMPASS threedimensional dose verification system,20 cases of upper and middle chest esophageal carcinoma on the VMAT plans for absolute dose and relative dose verification.Dose volume histogram (DVH) comparison treatment target,lungs,heart,and differences in the spinal cord irradiation dose and volume,and analyses γpass rate of GTV,CTV,PTV and organs at risks.Results The center dose of upper and middle chest esophageal carcinoma accurate rates were above 99％.Thoracic segment esophageal:GTV,PTV and organs at risks of γ pass rate above 97％.D95％ and Dmean of GTV,CTV and PTV were relatively undervalued within 3％.D1％ of spinal cord is 2.21％ overvalued.Left and right pulmonary V5 were slightly overvalued by about 0.5％,V10-D30,Dmean undervalued within 2％.In period of middle chest esophageal carcinoma:the gamma passed rate of GTV,CTV,PTV and organs at risks of above 97％,GTV,CTV,PTV D95％,Dmean were relatively undervalued within 2％.Spinal cord D1％ is 2.04％ overvalued.Left and right pulmonary V5-D30 to V10 as a trend of gradually to be underestimated,at 1.5％.Heart Dmean was undervalued by 2.68％.Conclusion VMAT technology is applicable in the chest esophageal carcinoma radiotherapy.

ObjectiveTo observe the clinical efficacy of combined irbesartan and alprostadil treatment of early diabetic nephropathy (DN).Methods A total of 120 patients with early type 2 diabetes of hospitalization were randomly divided into 3 groups: irbesartan group of 40 patients (150 mg, once per day)(group irbesartan),40 patients treated with alprostadil (physiological saline and alprostadi 10 μg))(group alprostadi) ,40 patients treated with alprostadil combined irbesartan (dose same as the other two groups)(combined group).All cases were observed for 4 weeks.Comparison of serum creatinine (Cr), blood urea nitrogen(BUN) ,24 hour urinary albumin(24hUAE) changes after treatment.ResultsAfter treatment 4 weeks 24hUAE of the three groups were significantly decreased (t = 2.07, t = 2.01 and t = 3.15, Ps ＜ 0.05) .The decrease of 24hUAE in the combined treatment group ([252.69 ± 33.56]mg/24h) was better than that in the irbesartan group([268.75 ± 34.42)](t = 2.11, P ＜ 0.05)and in the alprostadil group ([267.95 ± 30.75])mg/24h (t = 1.998, P ＜ 0.05) .No significant difference were observed between the Irbesartan and alprostadil group.During treatment, several patients affected by swell, uncomfortable in the alprostadil group, but improved after treatment.No other adverse effect was observed.Conclusion Alprostadil combined with irbesartan treatment of early diabetic nephropathy is an effective way.