Objective To establish a new multiplex-PCR assay to improve the detection rate of mutations in the DMD gene in Chinese patients. Methods A retrospective review of DMD deletion spectrum of 355 DMD patients with deletions all over the gene was performed. All deletions were confirmed by " one-step approach" diagnostic procedure and MLPA analysis. The exons with high frequency of mutations were identified to constitute the amplification system and the PCR conditions were optimized. Results Two new multiplex-PCR assays were established. Assay one was used to detect 10 exons including exon 5, 8, 17, 44, 45, 47, 49, 50, 51 and 52 of DMD gene, in two PCR sets. The theoretical detection rate would be 92% (326/355). Assay two was used to detect 5 exons including exon 12, 19, 35, 43 and 54, which could be used to screen additional 5% (17/355) deletion cases. The method was validated in other 22 DMD patients. Multiplex-PCR results were completely identical to the MLPA results in all 22 DMD patients. Conclusions The two multiplex-PCR assays were established based on the analysis of 355 Chinese DMD patients with gene deletions. It is believed that the new approach would be more applicable for deletion detection on the Chinese DMD patients since the DMD cases involved were from the whole country.

β- N-Acetyl- D-glucosaminidase ( NAGase, EC3.2.1.52) is a composition of the chitinases and cooperates with endo-chitinase and exo-chitinase to disintegrate chitin into N-acetylglucosamine. Pacific White Shrimp (P. vannamei) NAGase is involved in digestion and molting processes. Some pollutants in seawater affect the enzyme activity causing loss of the biological function of the enzyme, which affects the exuviating shell and threatens the survival of the animal. The effects of acetic anhydride on the enzyme activity for the hydrolysis of pNP-NAG have been studied. The results show that acetic anhydride can lead to reversible non-competitive inhibition at appropriate concentrations, and the IC50 is estimated to be 9.0 mmol/L. The equilibrium constants have been determined for acetic anhydride binding with the enzyme and/or the enzymesubstrate complexes. Inhibition kinetics of acetic anhydride on the enzyme has been studied using the kinetic method of the substrate reaction. The results suggest that at pH 6.2, the action of acetic anhydride on the enzyme is first quick equilibrium binding and then slow inhibition. The microscopic rate constants have been determined for inhibition and reactivation. The results show that k + 0 is much larger than k - 0, indicating the enzyme is completely inactivated at sufficiently large modificator concentration.

Objective To investigate the surgical efficacy for the treatment of ruptured anterior communicating artery aneurysms with different dome projections in acute phase via supraorbital lateral approach.Methods From January 2014 to March 2017,the clinical data of 79 patients with acute ruptured anterior communicating artery aneurysm with different dome projections in acute phase treated via supraorbital lateral approach in the First Affiliated Hospital of Soochow University were retrospectively analyzed.The operative efficacy was analyzed.The Glasgow outcome scale (GOS) score was used to evaluate the prognosis at 3-36 months after procedure.Results In the 79 patients,34 were superior dome projections,37 was inferior dome projections,and 8 were complicated dome projections.All the anterior communicating artery aneurysms were successfully clipped.The patients were followed up for 3-36 months after procedure.GOS showed good prognosis in 70 cases (88.6％),of which 26 were superior dome projections,36 were inferior dome projections,and 8 were complicated dome projections;9 suffered mild disability (11.4％),of which 8 were superior dome projections and 1 was inferior dome projection.There were no severe disability,persistent vegetative state,and death.The prognosis in patients with inferior dome projection was better than that of superior dome projection (97.3％ [36/37] vs.76.5％ [26/34]).The difference was statistically significant (χ2=5.19,P<0.05).Conclusions The lateral supraorbital approach is an effective option for treating ruptured ACoAA with different dome projections in acute phase.The prognosis in patients with inferior dome projection is better than that of superior dome projection.

Objective To detect the expression of Zbtb7 gene in Phorbol esters(PMA) induced differentiation of leukemia cell lines.Methods 50 nmol/L PMA was used to induce the differentiation of U937 and K562 cell lines.The expression of Zbtb7 was detected by real-time PCR in the cells after induction on day 0,1,3 and 5,respectively. Results The expression of Zbtb7 in leukemia cell lines was markedly enhanced after treated by PMA(P

Objective To study the effect of electroacupuncture (EA) combined with repetitive transcranial magnetic stimulation (rTMS) on endogenous neural atem cells, as well as learning and memory in rats with cerebral ischemia. Methods One hundred and twenty male Wistar rats were randomly divided into 5 groups: a normal group, a model group, an EA group, a rTMS group and an EA plus rTMS group. After establishing acute middle cer-ebral artery occlusion, the rats in the various experimental groups were treated with EA, rTMS or EA plus rTMS. The rats' ability to avoid an electric shock was observed using a jumping stand, Immunohistochemical techniques were used to measure the infarct area and the number of Brdu-positive cells. Results A small quantity of Brdu labelled cells was observed in the subgranular zone (SGZ) and the subventricular zone (SVZ) in rats in the normal group. In the EA, rTMS and EA plus rTMS groups, the numbers of Brdu labelled cells in the SVZ and SGZ was significantly higher than in the model group after both 7 and 14 days. The expression of Brdu around the SVZ and SGZ of the 3 treatment groups increased, especially in the EA plus rTMS group. At 7, 14 and 28 d there was significant improve-ment in the neural function and electric shock avoidance test scores in the EA, rTMS and EA plus rTMS groups com-pared with the model group,with the EA plus rTMS animals scoring best. Conclusion EA plus rTMS can promote the proliferation of nerve stem cells, facilitate neurological recovery and improve learning and memory in rats after cerebral ischemia.

To investigate the contamination conditions of Campylobacter spp.in duck production chain and its antimicrobial resistance,virulence gene distribution,samples were collected at the duck slaughterhouse according to GB 4789.9-2014.Triplex PCR assay was applied to identify the species of Campylobacter and the recovered Campylobacter strains were tested for the antimicrobial susceptibility against 8 kinds of antimicrobial agents using a broth microdilution method,the susceptibility results were determined according to the NARMS criteria (2011).Subsequently,4 virulence genes were detected by PCR method.The result showed that 187 Campylobacter isolates were obtained from 489 samples,including 160 C.jejuni,130 C.coli and 10 unidentified Campylobacter isolates.The total isolation rate of Campylobacter was 38.24％.The prevalence of Camnpytobacter before slaughtering,at depilation stage,evisceration stage and duck products was 76.33％,5.62％,24.00％,and 0％,respectively.The Campylobacter isolates were most frequently resistant to tetracycline (95.72 ％),followed by resistance to clindamycin(90.91％),the resistance rate of azithromycin (63.64％) was in the middle,the resistance rates of ciprofloxacin(31.02％),gentamicin(34.76％),nalidixic acid (37.43 ％),erythromycin (41.18 ％),chloramphenicol (41.18％) were relatively low.The multi-drug resistance was common among Campylobacter isolates with a rate of 72.19％.The prevalence of adhesion-associated gene cadF,flagellin gene flaA,invasion associated protein gene iamA,toxin regulation gene cdtB was 100％,80.75 ％,71.12％ and 94.65％,respectively.The results indicated that the Campylobacter contamination occurred in the slaughtering procedures of duck,and the antimicrobial resistance of Campylobacter isolates was relatively serious.In addition,the virulence-associated genes were detected widely among Campylobacter isolates.Therefore,the supervision of antimicrobial agents using at rearing stage should be strengthened,along with health management in duck production chain.

ObjectiveTo evaluate hybrid operation for arteriosclerosis obliterans (ASO) of the lower extremity.MethodsClinical data of 35 ischemic limbs in 32 ASO cases receiving hybrid operation from May,2007 to August,2009 were retrospectively analysed.The indications,clinical result,complications,perioperative mortality,vascular patency rate and limb salvage rate was evaluated.ResultsThirty-five ischemic limbs in 32 cases underwent hybrid operation. Procedures were successful in 94％ cases (33/35). The average postoperative ABI significantly increased from 0.49 ±0. 18 to 1.06 ± 0. 17 ( one day after surgery) or 0. 96 ± 0. 16 ( six months after therapy). One patient suffered limb amputation due to surgical failure, one case was complicated with cerebral infarction and the operation was terminated. No patient died in perioperative period. Twenty-six cases (28 ischemia limbs) were followed-up from 2 month to 28 months, the follow-up rate was 87％ (26/30). Vascular patency rate in 6 months after operation was 93％ (26/28), and limb salvage rate was 96％ (27/28). Two cases suffered from below-knee reocclusion 5 -6 months after therapy, and one of these two cases needed a limb amputation.ConclusionHybrid operation is the therapy of choice for multifocal lesions in arteriosclerosis obliterans of the lower extremities with a low risk and higher patency in short term.

To research the structure-activity relationship (SAR) of glycinamide-bearing compounds that used as inhibitors of dipeptidyl peptidase IV (DPP-IV), P32/98 and compound A were chosen as the leading compounds, heterocycles containing nitrogen atom were introduced to form amide, and different residues on a-position of carbonyl were designed. The nineteen designed compounds were synthesized by a simple route and were evaluated as inhibitors of DPP-IV. All of the structures were characterized by 1H NMR and HRMS. The preliminary SAR result was obtained.

?AlM: To observe the effect of Qingguang'an on elastic fiber, MMP-7, TlMP-1 in scarring area of filtration canal after glaucoma surgery through the four Qingguang'an effective groups and Qingguang'an granules, to discuss and compare their mechanism of action on scarring area of filtration canal.?METHODS:Four effective components of Qingguang'an were used in groups D, E, F, G and H after glaucoma surgery, compared with group A ( blank ) , group B (model) and group C ( MMC) to observe the effect of elastic fiber, MMP-7, TlMP-1 in scarring filtration canal.?RESULTS:Compared with the preoperative basic lOP and 2d , 1, 2, 4wk postoperative lOP of groups C, E and H, the lOP of three group rose up slower than other groups, and kept the lowest data at 28d. There was significant difference compared with the rest of A, B, D, F, G groups (P<0. 05). The area and density of elastic fiber in surgery group were significantly different with that of black control group ( P<0. 05 ), but there were no statistical differences between groups C and H, groups C and F, groups H and E (P>0. 05). The difference was statistically significant among other groups (P<0. 01).?CONCLUSlON:The scarring area of filtration canal after glaucoma surgery is the major reason which lead to the failure of surgery. Qingguang'an effective group 2, Qingguang'an granules and MMC could reduced the scar tissue by restrained the elastic fiber, TlMP - 1 and increased the MMP-7. By observing the experimental results that both Qingguang'an effective group 2 and Qingguang'an granules could restrained the scarring area of filtration canal, the effects were unbiased, Qingguang'an granules group is better than effective group 2.

With the development of cell separation technique, hepatocyte transplantation becomes a hot topic; however, the application is limited by donor deficiency and immunological rejection. Microencapsulated hepatocytes contribute to the promotion and application for liver cell transplantation, for which provide a large amount of high activity and good function of liver cells, in this paper, liver cell microencapsulation technology and its progress in applications were reviewed, providing prospective way for large-scale and high-active culture in vitro and long-term cryopreservation.