Bdellovibrio bacteria BDF-H16 was isolated from the gut of Carassius auratus gibelio with Aeromonas sobria.Its shape was ob- served by light microscopy,phase-contrast microscopy,electron microscopy and some of its biological characteristics were also studied.It was demonstrated that BDF-H16 was an gram-negative bacterium and had a bacilliform or arc bacilliform shape with a flagellum at one end.Its size was mostly 0.2?m～0.5?m?0.8?m～1.2?m.It had a wide prey area and could lyse all tested gram-negative bacteria and some gram-positive bacteria.The best lysis conditions to Escherichia coli were 6.75?10~9 cfu/mL of prey bacteria concentration,pH7.0～7.5,28℃.It could grow in the solid culture added 0.85%～5.00% NaCl and was inhibited by enrofloxacin and norfloxacin.

In the experiment, the production of plagues by Bdellovibrio bacteria in solid medium cultivation, the reproduction of Bdellovibrio bacteria in liquid medium cultivation and the attachment of Bdellovibrio bacteria to carrier were observed, which aimed to study the effects of enrofloxacin on the growth and at-tachment ability of Bdellovibrio bacteria BDF-H16. Results indicated that in solid medium cultivation, the production of plagues by Bdellovibrio bacteria BDF-H16 was inhibited by different concentrations (2 ?g/mL, 5 ?g/mL, 10 ?g/mL, 20 ?g/mL, 50 ?g/mL) of enrofloxacin and the inhibitory effects of enrofloxacin became stronger with the increase of the concentration of enrofloxacin. Similarly, in liquid medium cultivation, the reproduction of Bdellovibrio bacteria BDF-H16 was also obviously inhibited by different concentrations ofenrofloxacin and higher concentrations of enrofloxacin such as 10 ?g/mL, 20 ?g/mL, 50 ?g/mL had stronger inhibitory effects on the reproduction of BDF-H16. However, the growth tendency of Bdellovibrio bacteria BDF-H16 was not inhibited in 10 ?g/mL enrofloxacin. Additionally, when zeolite was added, enrofloxacin had also inhibitory effects on the numbers of Bdellovibrio bacteria BDF-H16 attached to zeolite. With the increase of the concentrations of enrofloxacin, the numbers of Bdellovibrio bacteria BDF-H16 attached to zeolite became smaller and smaller. However, the attachment rate of Bdellovibrio bacteria BDF-H16 to zeo-lite became higher under 2 ?g/mL-20 ?g/mL enrofloxacin. The results above showed that enrofloxacin had inhibitory effects on the plague production and reproduction of Bdellovibrio bacteria BDF-H16, but the at-tachment ability of Bdellovibrio bacteria BDF-H16 was strengthened in liquid medium cultivation with 2 ?g/mL-20 ?g/mL enrofloxacin and zeolite, and adding zeolite helped to reduce the adverse effects of en-rofloxacin on Bdellovibrio bacteria BDF-H16.

Six bacterial strains with malachite green decolorization ability were isolated from a sediment of aquaculture pond, and strain M6 was selected by further enrichment culture in nutrition broth with malachite green and decolorization rate comparison. The decolorization rate of strain M6 to malachite green was 97.14% in the conditon of 30?C and 150 r/min, and its morphology was observed by gram stain and electronmicroscopy, its physiological and biochemical characteristic was studied by ATB bacteria identification in-strument for identification of bacteria, and its 16S rDNA sequence was determined following PCR amplifi-cation, the sequence was aligned and the phylogenic tree was instructed with those bacterial strains of high identity with strain M6. In addition, its growth characteristics was also studied. The experimental results showed that strain M6 was gram negative and bacilliform with a flagellum at one end. Its size was 0.45 ?m ?0.84 ?m. Its colony produced on common agar plate appeared as round, light blue, dense, hard to choose; 16S rDNA sequence of strain M6 had high identity of 98%～99% with Pseudomonas sp. located in GenBank and strain M6 had the most close relative relation to Pseudomonas putida OW-16 (Locus number: DQ112328.1). Combined the results of the traditional morphological, physiological, biochemical character-istics and 16S rDNA sequence analysis, strain M6 was identified as Pseudomonas putida (Locus number: EU348741.1). Additionally, its growth curve in the condition of 30?C and 150 r/min was as follows: lag phase was 0～4 h, log phase was 4 h～64 h, stationary phase was 64 h～80 h, decline phase was after 80 h. Its best growth conditions were pH 7 and 30?C,and in the rotational speed of 50 r/min to 250 r/min. Its concen-tration increased with the increase in rotational speed.

To investigate the distribution of possible novel mutations from parkin gene in variant subset of patients with Parkinson's disease (PD) in China and explore whether parkin gene plays an important role in the pathogenesis of PD, 70 patients were divided into early-onset group and late-onset group; 70 healthy subjects were included as controls. Genomic DNA from 70 normal controls and from those of PD patients were extracted from peripheral blood leukocytes by using standard procedures. Mutations of parkin gene (exon 1-12) in all the subjects were screened by PCR-single strand conformation polymorphism (SSCP), and further sequencing was performed in the samples with abnormal SSCP results, in order to confirm the mutation and its location. A new missense mutation Gly284Arg in a patient and 3 abnormal bands in SSCP electrophoresis from samples of another 3 patients were found. All the DNA variants were sourced from the samples of the patients with early-onset PD. It was concluded that Parkin point mutation also partially contributes to the development of early-onset Parkinson's disease in Chinese.
DNA Mutational Analysis ; Exons ; Genotype ; Parkinson Disease/*genetics ; *Point Mutation ; Polymorphism, Single-Stranded Conformational ; Ubiquitin-Protein Ligases/biosynthesis ; Ubiquitin-Protein Ligases/*genetics

Objective To construct tissue engineering nucleus pulposus by culture of rabbit bone marrow mesenchymal stem cells (rBMSCs)-nucleus pulposus acellular matrix scaffold (NPAMS)complexes (rBMSCs-NPAMS).Methods Several NPAMS were prepared,and rBMSCs was seeded into NPAMS.The scaffolds and complex were detected by general observation,HE stai-ning,immunohistochemical,qRT-PCR,scanning electron microscopy.Results The scanning electron microscopy showed the seed cell in nucleus pulposus ECM-derived scaffold could adhesion and growth.The cell attachment and proliferation were observed by HE staining.Immunohistochemical examination with typeⅡ collagen showed positive results.qRT-PCR revealed the time-depend-ent of the mRNA expression of collagen Ⅱ,and which was smaller than positive control(P<0.01).The relative expression of Ag-grecan mRNA grew in a time dependent fashion,which was still lower than positive control(P<0.01).Scaffolds and normal nucle-us pulposus had no statistical significance of the compression load under the same displacement of the compression(P>0.05).Con-clusion Natural nucleus pulposus acellular matrix scaffold composite allogeneic bone marrow mesenchymal stem cells can be suc-cessfully built into tissue engineering nucleus pulposus.

Mutations in the parkin gene have recently been identified in familial and isolated patients with early-onset Parkinson disease (PD) and that subregions between exon 2 and 4 of the parkin gene are hot spots of deletive mutations. To study the distribution of deletions in the parkin gene among variant subset patients with PD in China, and to explore the role of parkin gene in the pathogenesis of PD, 63 patients were divided into early onset and later onset groups. Exons 1-12 were amplified by PCR, templated by the genomic DNA of patients, and then the deletion distribution detected by agarose electrophoresis. Four patients were found to be carrier of exon deletions in 63 patients with PD. The location of the deletion was on exon 2 (1 case), exon 3 (2 cases) and exon 4 (1 case). All patients were belong to the group of early onset PD. The results showed that parkin gene deletion on exon 2, exon 3 and exon 4 found in Chinese population contributes partly to early onset PD.
Exons/*genetics ; *Gene Deletion ; Parkinson Disease/*genetics ; Point Mutation ; Ubiquitin-Protein Ligases/*genetics

A pathogenic bacterial strain X1 was isolated from Siberian sturgeon (Acipenser baerii) suffering with bacterial septicemia. The 50% lethal concentration (LC50) of strain X1 was 5.62?105 CFU/mL, which showed that strain X1 was rather strong virulent to Acipenser baerii. Strain X1 was gram negative and 1.0 ?m～1.2 ?m ? 2.1 ?m～2.4 ?m in size with peritrichous flagella, and had ?-hemolytic activity on rabbit blood agar. By means of ATB expression identification and 16S rDNA sequence analysis, strain X1 was identified as Aeromonas hydrophila (locus number: EU669667), which was the closest relative to Aeromo-nas hydrophila strain ATCC35654 (locus number: X74676.1) with 99% homology. In addition, strain X1 was highly sensitive to cefoperazone and cravit, and intermediately sensitive to ten kinds of antibiotics in-cluding tobramycin, norfloxacin, sulperazone, kanamycin, gentamycin, fortum, vancomycin, neomycina, polymyxin B and lomefloxacin.

Objective: To investigate the mechanisms of electroacupuncture (EA) at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) in intervening diabetic gastroparesis (DGP) based on calcium-activated chloride channel. Methods: Forty Sprague-Dawley rats were randomly divided into four groups, including a normal control group (group A), a model group (group B), an EA group (group C) and a metoclopramide group (group D), with 10 rats in each group. A single intraperitoneal injection of 2％ streptozotocin (STZ) combined with 8-week high-glucose high-fat diet was used to establish a DGP rat model. After intervention, gastrointestinal propulsive rate was observed; the expression level of transmembrane protein 16A (TMEM16A) was examined by immunohistochemistry; the Ca2+ concentration in interstitial cells of Cajal (ICCs) was detected by immunofluorescence; and whole-cell patch-clamp technique was applied to detect the current intensity of calcium-activated chloride channel (ICaCC) in ICCs in gastric antrum. Results: After modeling, the blood glucose levels in group B, group C and group D were significantly increased compared with group A (all P<0.01); after intervention, compared with group B, the blood glucose levels in group C and group D were significantly decreased (P<0.05, P<0.01); the intra-group comparison of blood glucose level between after modeling and after intervention found significant difference only in group C (P<0.01). The gastrointestinal propulsive rates in group B, group C and group D were significantly different from that in group A (P<0.01 or P<0.05); the gastrointestinal propulsive rates were markedly higher in group C and group D than in group B (P<0.01, P<0.01). The expressions of TMEM16A in group B and group C were decreased compared with group A (P<0.01, P<0.05); the expressions of TMEM16A in group C and group D were increased compared with group B (P<0.01, P<0.05). The fluorescence intensity of Ca2+ was significantly lower in group B than in group A (P<0.01); the fluorescence intensity of Ca2+ was significantly higher in group C and group D than in group B (P<0.01, P<0.05). ICaCC in ICCs in group B was significantly decreased compared with group A; ICaCC in group C and group D were increased compared with group B. Conclusion: EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) can significantly improve gastrointestinal motility in DGP rats by up-regulating the ICaCC in ICCs.

Mutations in the parkin gene have recently been identified in familial and isolated patients with early-onset Parkinson disease (PD) and that subregions between exon 2 and 4 of the parkin gene are hot spots of deletive mutations. To study the distribution of deletions in the parkin gene among variant subset patients with PD in China, and to explore the role of parkin gene in the pathogenesis of PD, 63 patients were divided into early onset and later onset groups. Exons 1-12 were amplified by PCR, templated by the genomic DNA of patients, and then the deletion distribution detected by agarose electrophoresis. Four patients were found to be carrier of exon deletions in 63 patients with PD. The location of the deletion was on exon 2 (1 case), exon 3 (2 cases) and exon 4 (1 case). All patients were belong to the group of early onset PD. The results showed that parkin gene deletion on exon 2, exon 3 and exon 4 found in Chinese population contributes partly to early onset PD.
Adult ; Aged ; Exons ; genetics ; Female ; Gene Deletion ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; Point Mutation ; Ubiquitin-Protein Ligases ; genetics

The aim of the present study is to investigate the differences in cardiac function, and the expression and activity of calcium regulatory proteins between rabbit systolic heart failure (SHF) and diastolic heart failure (DHF) models. New Zealand white rabbits were randomly divided into three groups: sham operation (SO) group, DHF group (receiving abdominal aortic constriction) and SHF group (receiving aortic valve destruction and abdominal aortic constriction). The cardiac function was detected by echocardiographic and hemodynamic assays. The mRNA expression levels of sarcoplasmic reticulum Ca(2+) ATPase 2a (SERCA2a) and phospholamban (PLB) were evaluated by RT-PCR. The protein expression levels of SERCA2a, PLB, phosphoserine 16-PLB (pSer-16-PLB) and protein kinase A (PKA) were evaluated by Western blot, and the phosphorylation status of PLB was determined by the ratio of pSer-16-PLB protein level to that of PLB. The activity of SERCA2a was measured through inorganic phosphate. The activity of PKA was measured by gamma-(32)P ATP-binding assays. Compared with SO group, there were significantly increased ventricular wall thickness, raised left ventricular end diastolic pressure (LVEDP), reduced diastolic function in DHF group (P<0.05 or P<0.01), and significantly increased ventricular cavity size and LVEDP, reduced systolic function in SHF group (P<0.05 or P<0.01). The expression levels of SERCA2a in DHF and SHF groups were lower than that in SO group (P<0.05), while the expression and activity of PKA in DHF and SHF groups were higher than that in SO group (P<0.05 or P<0.01), and there was no significant difference between DHF and SHF groups. The expression levels of PLB and pSer-16-PLB as well as the phosphorylation status of PLB and activity of SERCA2a in SHF group were lower than those in DHF and SO groups respectively. Posing a contrast, the phosphorylation status of PLB and activity of SERCA2a in DHF group were higher than that in SO group (P<0.05). These results indicate that the SHF and DHF models were successfully established, and there are some differences in the expression and activity of calcium regulatory proteins between two models.
Animals ; Calcium-Binding Proteins ; metabolism ; Disease Models, Animal ; Heart Failure, Diastolic ; metabolism ; Heart Failure, Systolic ; metabolism ; Rabbits ; Sarcoplasmic Reticulum ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism