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Articles by M. Zhang
Total Records ( 9 ) for M. Zhang
  Y Yuan , J Tan , Y Wang , C Qian and M. Zhang
 

Chitosan (CS), a biocompatible and biodegradable material, can act as a non-viral delivery vehicle with low toxicity. In this study, plasmid DNA (pDNA) and siRNA were encapsulated in CS nanoparticles (NPs) to prepare CS–DNA and CS–siRNA NPs using a complex coacervation process. The CS–DNA particle size was within the range of 180–370 nm with a surface charge ranging from 0 to 18 mV at pH 5.5. The stability of pDNA in CS–DNA was investigated by pDNA release study and DNase I protection assay. The release of pDNA from NPs was studied in pH 7.4 phosphate-buffered saline at 37°C and the CS–DNA NPs could delay the DNA release. Results of DNase I protection assay showed that CS–DNA NPs could protect the encapsulated pDNA from nuclease degradation. In the transfection study, it was found that the transfection efficiency in vitro was dependent on the molecular weight, charge ratio, and DNA concentration of the CS–DNA NP as well as the type of cell transfected. Moreover, the morphology of HeLa cells transfected with CS–siRNA complexes was studied using atomic force microscopy. The results suggest that CS may be more capable than liposome in delivering siRNA to target cells. In summary, our analysis suggests that pDNA and siRNA can be encapsulated in CS NPs without being damaged.

  Z. Pei , X. Chen , C. Sun , H. Du , H. Wei , W. Song , Y. Yang , M. Zhang , W. Lu , R. Cheng and F. Luo
 

Aims

To examine single nucleotide polymorphisms in the protein tyrosine phosphatase N22 gene (PTPN22) and to study their association with Type 1 diabetes in a Chinese cohort.

Methods

Three hundred and sixty-four young patients with Type 1 diabetes and 719 healthy children were included in this case-controlled study. The genotypes of rs1217385, rs2488457 (-1123C>G), rs1217414, rs1217419, rs3765598 and rs2476601 (1858C>T) in the PTPN22 gene were determined using the SNaPshot method. Alleles, genotypes and haplotype frequencies were compared between patients with Type 1 diabetes and healthy control subjects. The association between single nucleotide polymorphisms and clinical traits/autoantibody status was also analysed.

Results

The single nucleotide polymorphism, rs1217419, located in the second intron of the PTPN22 gene was associated with Type 1 diabetes (odds ratio 1.5, 95% CI 1.14-1.97, P = 0.003). An additional single nucleotide polymorphism, rs1217385, was also associated with Type 1 diabetes; however, the association was secondary to that of rs1217419. The previously reported single nucleotide polymorphism that is associated with Type 1 diabetes (-1123G>C) had only marginal association with Type 1 diabetes in our study. A marginal association was also identified between -1123G>C and glutamic acid decarboxylase autoantibody positivity in patients with Type 1 diabetes. There was no association between the single nucleotide polymorphism 1858C>T and Type 1 diabetes in our studied cohort.

Conclusions

Our study confirmed that PTPN22 is a gene that contributes to Type 1 diabetes susceptibility. The primary association occurs with single nucleotide polymorphism rs1217419 and there is clear heterogeneity of the association between PTPTN22 polymorphisms and Type 1 diabetes in a Chinese population compared with other populations.

  Y Hirabayashi and M. Zhang
  No Description
  W. Wang , L. Wang , J.N. Li and M. Zhang
  Nine bacterial strains (LA1, LA2, LA4, JA1, CA3, CA4, BA1, BA18 and EA9) were isolated from different aquaculture animals with haemorrhagic septicemia and then they were identified as A. hydrophila (LA4, JA1, CA3 and BA1), A. sobria (LA1 and EA9), A. caviae (BA18) and A. veronii (CA4 and LA2), respectively by morphological and biochemical characterization. All isolates were found to be pathogenic to experimental zebrafish (Danio rerio) by artificial infection test. The outer membrane protein Aha1 is a major adhesin of A. hydrophila and also highly conserved in different serotypes of A. hydrophila. In order to ascertain the conservation of aha1 protein among mesophilic motile aeromonads, full length aha1 gene from all isolates was detected, cloned and sequenced. As the results show, the aha1 genes were amplified in all strains and the ORF size of the aha1 gene from A. hydrophila and other phenotypic species of aeromonas isolates was 1,068 and 1,038 bp, respectively. Four Anhui A. hydrophila isolates and six A. hydrophila reference strains formed a cluster together with 91.4-99.7% nucleotide identity and 91.9-99.7% amino acid identity of the aha1 gene. Five Anhui other phenotypic species isolates formed another cluster, they shared 79.5-81.1% nucleotide identity and 79.6-81.6% amino acid identity of the aha1 gene compared with A. hydrophila and the major sequence variations were observed between amino acids 85-134, 176- 227, 243-263, 280-295 and 321-336.
  X.M. Song , S.S. Lu , M. Wang , Q.Y. Li , D. Li , X.G. Yang , Y.Q. Lu , M. Zhang and K.H. Lu
  This study evaluated the effects of Glutathione (GSH) supplemented in the semen extender during the buffalo sperm sorting process on sperm quality, embryonic development after IVF and pregnancy rate after AI. The percentage of sperm motility, plasma membrane integrity and DNA fragmentation were detected by flow cytometry or by microscopy in stained, sorted and frozen semen treated with or without 0.75 mM GSH during the flow sorting procedure. The cleavage and blastocyst rates were examined at day 2 and 6-8 after IVF with frozen semen treated with or without 0.75 mM GSH. Pregnancy diagnosis was determined by transrectal palpation at 90 day after AI with frozen semen treated with or without 0.75 mM GSH. The percentage of sperm with Progressive Motility (MP, %) was significantly higher (p<0.05) in sorted semen supplemented with 0.75 mM GSH than that in the control. The percentages of moribund, dead and Phosphatidylserine (PS) translocated sperm detected by flow cytometry were significantly decreased (p<0.05) in frozen semen supplemented with GSH compared to the control. Higher blastocyst and pregnancy rates (p<0.05) were found after IVF and AI with frozen sperm treated with 0.75 mM GSH than that in the control group. In conclusion, addition of 0.75 mM GSH to the semen extenders (stained, sorted and frozen) during the sperm sorting process can improve sperm quality in vitro embryonic development and in vivo fertility after AI thus indicating potential for commercial application in buffalo sperm sorting.
  X.B. Zheng , X.M. Cen , L.X. Wang , G.Y. Xin , X.H. Fu , P. Liu , G.H. Li , M. Zhang , S.S. Lu and K.H. Lu
  Nanog is one of important transcription factors to maintain characteristics of pluripotent stem cells. The present study was to clone Nanog of Capra hircus to express His-Nanog protein in E. coli BL 21 cells and further to purify it. The total RNA was extracted from primordial genital ridge tissues of a fetal lam and by means of RT-PCR, Nanog gene was amplified which was subcloned to pET32a to construct its prokaryotic expression vector. Confirmed by restrictive endonuclease digestion and DNA sequencing, the recombinant plasmid was transformed into E. coli BL21(DE3) and His-Nanog fusion protein was expressed by the induction of IPTG and identified with SDS-PAGE analysis. Under denaturing condition, the His-Nanog protein was purified by using Ni-NTA resin and verified by Western blotting assay. The results showed that The Open Reading Frame (ORF) of Nanog gene in Capra hircus is composed of 903 nucleutide acids, coding 320 amino acids; SDS-PAGE assay showed that His-Nanog fusion protein was efficiently expressed in form of inclusion bodies in E. coli BL21 (DE3) inclusion bodies were solubilized in 6 mol L-1 GuHCl, His-Nanog fusion protein with higher purity was purified by using Ni-NTA resin; Western blotting assay showed that the purified His-Nanog could bind to anti-His tag antibody specifically indicating the expected immunogenicity. This recombinant protein could be used directly to prepare polyclonal or monoclonal anti-Nanog antibody which will lead to study Nanog’s function or characteristics of pluripoten stem cells (such as iPS cells) in Capra hircus.
  X Zhang , L Wang , A Lu and M. Zhang
 

Objective: To summarize the clinical characteristics of idiopathic pulmonary haemosiderosis (IPH) to explore the aetiopathogenesis, risk factors, diagnosis and experiences in therapy of IPH. Methods: The documents of 28 IPH cases, who were hospitalized in Children’s Hospital of Fudan University between February 1989 and June 2009 were reviewed. Results: (i) fifteen cases were males and 13 were females, and 88.5% of the cases had first onset under the age of 10 years; (ii) the triad occurred in 57.1% cases; (iii) radiographic features of IPH including diffuse alveolar-type infiltrates, ground glass attenuation, interstitial reticular and micronodular patterns; (iv) haemosiderin-laden macrophages were found in 60.7% of the cases;(v) the trend of positive correlation was found between the severity of ventilatory restrictive pattern and the disease courses (r = 0.229, p = 0.237); and (vi) glucocorticosteroids can control the symptoms. Conclusion: (i) the clinical presentations are not classical. If long-term anaemia exists without reason, this case must be considered; (ii) corticosteroid can control the symptom; and (iii) IPH may be associated with the imbalance of immune system.

  X. j Cai , L Chen , L Li , M Feng , X Li , K Zhang , Y. y Rong , X. b Hu , M. x Zhang , Y Zhang and M. Zhang
 

Adiponectin is an important antiatherogenic adipocytokine that inhibits inflammation, insulin resistance, and oxide stress. Inflammation in the vascular adventitia is a crucial factor in the pathogenesis of atherosclerosis. Adventitial fibroblasts (AFs) can proliferate, divide into myofibroblasts, and migrate to the intima to become a new component of atherosclerotic plaque under inflammation and atherosclerosis. We investigated whether adiponectin might prevent AFs from proliferating, migrating, and transforming into myofibroblasts. Cultured AFs were stimulated with lipopolysaccharide (LPS) in the presence or absence of adiponectin. Methyl thiazolyl tetrazolium assay and migration and scratch-wound assays demonstrated that adiponectin reduced the AF proliferation and migration induced by LPS, respectively, whereas treatment with AdipoR1 small interfering (si) RNA (siAdipoR1), AMP-activated protein kinase (AMPK) siRNA (siAMPK), and an AMPK inhibitor reversed the effect. Immunocytochemistry and Western blot revealed that adiponectin reduced the transition of AFs to myofibroblasts, and treatment with siAdipoR1, siAMPK, and the AMPK inhibitor increased the transition. RT-PCR, Western blotting, and nitric oxide (NO) assay showed that adiponectin reduces induced NO synthase (iNOS) and nitrotyrosine expression and NO and ONOO production induced by LPS. Treatment with siAdipoR1, siAMPK, and the AMPK inhibitor significantly attenuated adiponectin-induced phosphorylation of AMPK and its downstream target acetyl-coenzyme A carboxylase and up-regulated iNOS mRNA and protein expression, which resulted in a marked increase of NO and ONOO production. In apolipoprotein E-deficient mice, immunohistochemistry of treated vascular adventitia showed that both iNOS expression and ONOO production could be reversed with an adenovirus-adiponectin vector. Taken together, these results suggest that adiponectin reduces LPS-induced NO production and nitrosative stress and prevents AFs from proliferating, transforming to myoflbroblasts, and migrating to the intima, thus worsening atherosclerosis, by inhibiting the AdipoR1-AMPK-iNOS pathway in AFs.

  L.V. Raymond , M. Zhang and S.M. Roknul Azam
  Edible coatings, thin layer formed on the surface of a product, have been used to preserve fresh-cut fruits and vegetables since they can improve quality by affecting respiration and moisture loss. Green pepper slices were subjected to chitosan coating treatment achieved by dipping, afterwards stored at 5°C for a period of 15 days. The effect of various chitosan concentrations (0%, 0.5%, 1.0% and 2% (w/v) chitosan) on some physico-chemical and microbial characteristics of the slices was subsequently analyzed during storage. The data indicated that the performance of chitosan treatments was better than that of control. The decrease of surface lightness was delayed by chitosan treatments; and the surface green colour was kept under marginal changes compared to control. On the other hand, a greater reduction of fungal incidence, carbon dioxide concentration and electrolyte leakage of the chitosan treated samples was observed with increased chitosan concentration treatment. Microbiological evaluation revealed that total viable cells count decreased with increasing chitosan concentration. This correlated well with the external changes which were affected to a lesser extent in the 2% chitosan compared to the control; in addition, delaying of changes was significantly chitosan concentration dependent.
 
 
 
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