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.

To optimize the process of hydrogenation reduction in the synthesis of apremilast (APST), 3-nitrophthalic anhydride (4) and (S)-1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethanamine-(S)-2-acetamido-4-methylpentanoate (7) were used as starting materials to synthesize (S)-2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl)-4-nitroisoindoline-1,3-dione (8) by amination.Then compound 8 was reduced to (S)-4-amino-2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethyl) isoindoline-1,3-dione (9) with ammonium formate as hydrogen source and palladium hydroxide as catalyst.Finally, apremilast was obtained by the acetylation reaction with acetic anhydride.The structure of the products were verified by optical rotation, 1H NMR, 13C NMR, MS and elemental analysis.And the total yield of three steps was increased to 67.0%.The improved reduction process can avoid the special reaction of hydrogenation and pressurization, and reduce the safety risk and production costs with high commercial value.

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.

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 investigate the mechanism of Deinococcus radiodurans pprI gene in enhancing mice radioresistance to γ-rays by transfecting it in vivo.Methods The male Kunming mice were randomly divided into control group,irradiated group,pCMV-HA transfected group and pCMV-HA-pprI transfected group.The pCMV-HA-pprI plasmid contained pprI gene was injected into the muscle of mice which were exposed to total 6 Gy of γ-ray irradiation.After injection,the in vivo gene electroporation technology was used to transfect the pprI gene into the cells,and Western blot was used to identify the PprI protein,mammalian homolog protein Rad51 corresponding to recA gene downstream of pprI,and protein Rad52.Results In the muscle of the mice of transfected pCMV-HA-pprI group,the protein PprI expressed significantly at 1 d post-irradiation,but there was no expression of pprI gene 7 d post-irradiation and in other groups.In the mice of transfected with pCMV-HA-pprI,the expression of Rad51 protein was significantly increased in the lungs at 1,7 and 14 d post-irradiation,and significantly increased in the liver at 1 and 28 d post-irradiation and increased in the kidneys at 1 and 14 d post-irradition.However,there was no obvious change of Rad52 protein expression in the lungs and livers of mice in all groups.Conlusions The prokaryotic gene pprI could act on the mammalian homologisation analogues rad51 gene downstream of recA gene and then increase the expression level of protein Rad51 which results in the enhancement of radioresistance.

Objective To evaluate the protective effect of sinomenine (SIN) on renal ischemia/reperfusion (I/R) in mice. Methods In the experiment one, 12 C57BL/6 mice were randomly divided into 2 groups: SIN group (mice were injected with 200 mg/kg SIN by tail vein) and control group (mice were injected with equal volume of saline). Six and 24 hs later, the serum was collected and the contents of alanine aminotransferase (ALT) and creatinine (SCr) were determined. In the experiment two, C57BL/6 mice were randomly divided into 3 groups: sham-operated (SO) group, SIN group (mice were injected with 200 mg/kg sinomenine just before ischemia induction) and saline group (mice were injected with equal volume of saline at the same time). At the 6th h after reperfusion, the sera and renal samples subject to IR injury were collected. The SCr and BUN levels in serum were determined and renal histological changes were also examined. The apoptosis of renal tubular epithelial cells was measured by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay. The infiltration of F4/80 positive macrophages was measured by using immunohistochemistry and that of neutrophils with myeloperoxidase (MPO) kits. The mRNA expression of tumor necrosis factor (TNF)-α, chemokine CXC ligand (CXCL)-10, intercellular adhesion molecule (ICAM)-1 and IL-17 was detected by using real-time reverse transcription PCR. The activation of transcription factor NF-κB was measured by using Western blotting. Results In the experiment one, there was no significant difference in ALT and SCr between the two groups at 6 or 24 h. In the experiment two,levels of SCr and BUN were lower in SIN group (P＜0. 05 or P＜0. 01 ), histological damage was milder (P＜0. 01 ), and apoptosis rate of renal tubular epithelial cells apoptosis was lower than in saline group (P＜0. 05). The infiltration of macrophages, neutrophils and the mRNA expression of TNF-α, CXCL-10, ICAM-1 and IL-17 in the renal tissue in SIN group were reduced as compared with saline group (P＜0. 05 or P＜0. 01 ). The activation of NF-κB in SIN group was significantly downregulated as compared with saline group. Conclusion SIN can ameliorate the renal IR injury without hepatic or renal toxicity, which is associated with inhibition of acute inflammatory response induced by reperfusion.

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.

Objective To evaluate the effects of Alprostadil combined with Enalapril on high sensitivity C-reactive protein and Cystatin in patients with early diabetic nephropathy.Methods One hundred and fifteen cases of outpatients were randomized into 3 groups.Thirty-seven cases were assigned to Alprostadil group and treated with Alprostadil 10 μg + NS 100 ml,iv,qid;Thirty-eight cases were assigned to the Analapril group and treated with Enalapril 5 mg bid;Forty cases were assigned to the combined treatment group and treated with Alprostadil 10 μg + NS 100 ml,iv qid and Enalapril 5mg bid.All patients were observed for twelve weeks.Changes before and after treatment in the blood pressure,plasma glucose,glycolated hemoglobin (HbA1 c),24 hours urinary albumin (24 hUAE),high-sensitive C-reactive protein(hs-CRP) and Cystain C( Cys C) were observed and compared between the three groups.Results After treatment,hs-CRP,CysC and 24 h UAE were significantly improved in the three groups compared with baseline levels( P ＜ 0.05).In the combined treatment group,hs-CRP,CysC and 24 h UAER had a more significant improvement than the other two groups (P ＜0,05 ).Conclusion Alprostadil combined with Enalapril is a clinically effective strategy in the treatment of early diabetic nephropathv and can reduce the levels of CysC and hs-CRP.