OBJECTIVETo study the protective effect of puerarin on MPP(+) -induced SH-SY5Y cells by chaperone-mediated autophagy (CMA).

METHODThe Parkinson's disease cell model was established by injuring SH-SY5Y cells with 1 mmol x L(-1) MPP+. The CCK-8 staining was adopted to detect the effect the puerarin of different concentrations on the survival rate of MPP(+)-induced SH-SYSY cells. The autophagosome formation was observed under transmission electron microscope. The AO staining showed the changes in the lysosome activity. RT-PCR was used to detect the changes in Lamp2a and Hsc70 mRNA expressions. The western blotting was adopted to test the expressions of Lamp2a, Hsc70 and alpha-synuclein protein in cells.

RESULTWithin the concentration range of 12. 5-50.0 micromol x L(-1), the pretreatment with puerain for 30 minutes could protect the injury of MPP+ in SH-SY5Y cells, and showed a certain dose-effect relationship. The AO staining and electron microscope showed the effect of puerain within the concentration range of 12.5-50.0 micromol x L(-1) on 1 mmol x L(-1) MPP(+)-induced SH-SY5Y cells; autophagosomes emerged in cells, and increased along with the rise in the puerarin dose. The results of the flow cytometry revealed that 50.0 micromol x L(-1) of puerarin could protect against the increase of the ROS level in 1 mmol x L(-1) MPP(+) -induced SH-SY5Y cells and prevent the oxidative injury. The results of RT-PCR and western blotting indicated that puerain within the concentration range of 12.5-50.0 micromol x L(-1) alleviated the MPP(+)-induced SH-SY5Y cell injury, and inhibited the accumulation of alpha-synuclein proteins in MPP(+) -induced SH-SY5Y cells by up-regulating Hsc70, Lamp2a mRNA and protein level.

CONCLUSIONPuerarin could protect against the MPP(+) -induced cell injury, whose protective mechanism may be related to the chaperone-mediated autophagy pathway of interventional molecules.

Autophagy ; drug effects ; genetics ; HSC70 Heat-Shock Proteins ; genetics ; Humans ; Isoflavones ; pharmacology ; Lysosomal-Associated Membrane Protein 2 ; genetics ; Molecular Chaperones ; genetics ; Parkinson Disease ; drug therapy ; genetics ; Phagosomes ; drug effects ; genetics ; Piperidines ; pharmacology ; Pyrazoles ; pharmacology ; Tumor Cells, Cultured ; Up-Regulation ; drug effects ; genetics

Jin's three-needle technique is named after Professor JIN Rui. The connotation of Jin's three-needle technique is summarized by his followers through long-term teaching and clinical practice. His principle and method of point combination are based on syndrome differentiation of acupuncture. And his unique needling technique and treating principle of mental tranquilization also play an important role in his clinical practice.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Needles

China ; History, 20th Century ; History, 21st Century ; Pathology ; Periodicals as Topic ; history

NDV strain ZJ1 strain , a highly virulent NDV strain, has been prevalent among the waterfowls in China mainland in the past years. Multi-basic amino acid sequence distribute in the protease cleavage site of F protein of this strain. Recombinant expressing plasmid pCI-FT, was generated by converting multi-basic amino acid sequence of 112, 115, 117 of the protease cleavage site of F_ 0 protein, to the non-basic amino acid sequence characteristic of avirulent NDV strain. The result from co-expression of mutant or parental F protein with homologous HN protein in COS-1 cells revealed that both mutant and parental F protein had fusion activity. The result from co-expression of mutant or parental F protein with homologous HN protein in CEF cells showed that the cleavage activity of mutant F protein was significantly reduced. The study built a foundation for mutagenesis of amino acid sequence of the protease cleavage site of F_ 0 protein at the full-length cDNA clone level, study on factors contributing to virulence and construction of candidate vaccine strain, and so on.

A high productive poly ?-glutamic acid(?-PGA) strain PGA-N in a culture medium containing no L-glutamine was isolated from fermentation products.With the following identifications of colony mor-phology,physiological and biochemistry experiments,and genetics,the strain PGA-N was classified as a Bacillus licheniformis.According to the product environment,the base culture medium having no L-glutamine was simulated.In order to enhance the production of the strain PGA-N,the fermentation condi-tions,such as carbon source,nitrogen source,were optimized and the ?-glutamic acid production reached 5.16 g/L after getting the optimum formulation of this culture medium.PGA-N was mutagenized with com-bination of NTG and UV.A mutant PGA-N-C10 was screened which PGA production was increased from 5.16 g/L to 8.82 g/L.The study also investigated the effects of agitation speed on the cell biomass,?-PGA production and the ?-PGA molecular weight.The ?-PGA yield of PGA-N-C10 was as high as 11.00 g/L when the agitation speed was 400 r/min.

Objective To explore protective effect of Heme oxygenase-1(HO-1) on irradiation-induced endothelial cell apoptosis.Methods Human endothelial cell line EA.hy926 were administered with or without HO-1 activator Copp and/or HO-1 inhibitor Znpp,respectively.Then,cells were treated with or without 8 Gy radiation.The HO-1 protein expression of cells were assessed with Western blotting and apoptosis of cells treated with irradiation were evaluated with flow cytometry.Moreover,cytochrome C releasing into cytosol were also determined by Western blotting. Results In PBS+R group,HO-1 protein expression of EA.hy926 was low posterior to irradiation.When cells were preconditioned with Copp and/or Znpp,then recieved with 8Gy irradiation,the HO-1 protein expression of EA.hy926 increased significantly in comparision with the PBS+R group(P

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.

An extremely thermoacidophilic isolate K4-1 was obtained from an acidic hot spring in Teng- chong Rehai, Yunnan province. Morphology, growth characteristics, utilization of carbon compounds, en- ergy sources and 16S rRNA gene sequence of K4-1 were studied. Cells of K4-1 are irregular cocci with monotrichous flagella. The strain grew aerobically in either a lithotrophic or a heterotrophic mode. Growth on elemental sulfur occurred through oxidation of sulfur. It grew optimally at 75?C and pH 3.5. On the basis of 16S rRNA gene sequence similarity, strain K4-1 was shown to belong to genus Sulfolobus, being related to the type strains of genus Sulfolobus (86.6%~94.3% similarity), and being most closely related to strain Sulfolobus tengchongensis RT8-4 (98.9% similarity). The GenBank accession number of strain K4-1 16S rRNA gene sequence is EU729124.

OBJECTIVETo evaluate the efficacy and safety of "Three-Typed Syndrome Differentiation" (TTSD) in treating type 2 diabetes mellitus patients.

METHODSA systematic review and meta-analysis was done based on the clinical diabetes treatment literature of the "TTSD". Overseas databases like the PubMed/MEDLINE, EMBASE, Cochrane Library and Cochrane Central Register of Controlled Clinical Trials, and China databases like China Biology Medicine Disc (CBM), Chinese national Knowledge Infrastructure (CNKI), Wanfang database, and VIP database, without limitation on language, were included with the time limitation from Jan 1982 to Dec 2012 by retrieval of relative original clinical research articles.

RESULTSNineteen articles where contains 1,840 diabetes patients were obtained, in which no adverse reactions were reported. Of these, 14 literatures involved the effect of fasting blood glucose (FBG), 10 involved that of postprandial 2-h blood glucose (P2hBG), and 19 involved the overall efficacy based on the national Chinese medicine (CM) diagnosis and treatment standard of diabetes. All the meta-analysis results prefer to the "TTSD" groups (CM+Western medicine Based on TTSD). The results show that, beside the efficacy of Western medicine, the concentrations of FBG and P2hBG in "TTSD" groups continue to drop with statistical significance. For "TTSD" groups, the FBG subsequently dropped 1.03 mmol/L, 95%CI [1.24,0.82] P <0.00001), the P2hBG subsequently dropped 1.09 mmol/L, 95% CI [1.61, 0.57] (P <0.0001), and the overall efficacies benefit 3.46 times those of Western medicine alone, 95% CI [2.67,4.48] (P <0.00001).

CONCLUSIONSThe CM by the diagnosis and treatment of type 2 diabetes based on TTSD might be safe and effective, and could better improve both blood glucose and the overall status of patients, including symptoms.

Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Fasting ; blood ; Humans ; Medicine, Chinese Traditional ; Postprandial Period ; Syndrome ; Treatment Outcome

Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers.The fragments,amplified with primer Ⅰ to Ⅶ,were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain.The eukaryotic expression vector pCI-L was constructed by subcloning the fragments,amplified with the primer Ⅴ,Ⅵ and Ⅷ,into the expression vector pCI-neo.The full-length cDNA clone,pNDVZJI,with three helper plasmids,pCI-NP、pCI-P and pCI-L,were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase.After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF)flock,The NDV of ZJI strain was rescued successfully,which laid a good foundation for the further related research.