Calbindin 2 ; metabolism ; Heart Neoplasms ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; MyoD Protein ; metabolism ; Myogenin ; metabolism ; Rhabdomyosarcoma, Embryonal ; metabolism ; pathology ; surgery

Objective To investigate the resistant mechanism of imipenem-resistant K. pneumoniae.Methods The minimal inhibitive concentrations (MICs) of the antimicrobial agents were determined by Etest.Isoelectric focusing electrophoresis (IEF),plasmid extraction,conjugation, transformation,PCR amplification,cloning and sequencing were carried out for analyzing the encoding gene of ?-1actamases.Results Three kinds of ?-1actamases were detected with pIs of 7.2,6.7,and 5.4.in a clinical strain of K.pneumoniae.These ?-1actamases were TEM-I (pI,5.4),SHV-12 (pI,8.2) and KPC-2 ( pI,6.7 ) confirmed by sequencing of the PCR products.Only one band of ?-1actamase with pI 6.7 was displayed in the transformant.A 1500 bp segment,which contained the KPC-2 gene confirmed by nucleotide sequence analysis,was cloned from a 60 000 bp plasmid of the transformant.Conclusion The strain of K.pneumoniae resistant to imipenem produces a plasmid-mediated carbapenemase KPC-2 which belongs to Bush group 2f,class A ?-1actamase.

Objective To investigate the prevalence,genotype and epidemiology of plasmid- mediated AmpC enzyme of Escherichia coli and Klebsiella pneumoniae.Methods A total of 67 clinical isolates of nonrepetitive cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae collected by Fuzhou General Hospital and Fujian Provincial Hospital during a period of Sept.2004 to Mar.2005 were detected by three-dimensional extract test for AmpC enzyme,and PCR for AmpC enzyme and other ?-lactamase gene amplification and DNA sequencing were carried out for genotype of ?-lactamase.Plasmid transformation experiment was used to study the transfer of cefoxitin resistance.The homology of the isolates was determined by ERIC-PCR fingerprinting.Results At two hospitals in Fuzhou,the prevalence of plasmid-mediated AmpC enzyme among cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae were 16.7% and 10.5%, 8.0% and 0,respectively.Two isolates of Klebsiella pneumoniae produced DHA-1 plasmid-mediated AmpC enzyme,and 4 isolates of Escherichia cob and one strain of Escherichia coli produced CMY-2 and CMY-22 plasmid-mediated AmpC enzyme respectively.Furthermore,5 strains of Escherichia coli with CMY AmpC enzyme were also found simuhaneously to produce TEM-144,CTX-M-27,CTX-M-14 and TEM-1 ?-lactamase respectively.Three strains of Escherichia coli and one isolate of Klebsiella pneumoniae could transfer cefoxitin resistance to acceptant bacillus.ERIC-PCR fingerprinting reveals 2 strains of Klebsiella pneumoniae came from same clone,but 5 strains of Escherichia coli came from different clones.Conclusions The clinical isolates of Klebsiella pneumoniae producing DHA-1 plasmid-mediated AmpC enzyme and Escherichia coli producing CMY-2,CMY-22 plasmid-mediated AmpC enzyme are found in Fuzhou.CMY-22 AmpC enzyme and TEM-144 ?-lactamase are the first reported in the world,GenBank accession number: DO256079,DO256080

OBJECTIVETo investigate the changes in cerebral blood flow in patients with maxillary defect treated with prosthesis insertion.

METHODSThirty patients with maxillary defect receiving obturator prosthesis insertion were enrolled with another 30 subjects without dentition defect as the control. The cerebral blood flow rate was recorded before and at 5 and 10 min during mastication, and the results were analyzed statistically.

RESULTSThere was no significant difference in Vs, Vd or Vm between the two groups at the time points for measurement.

CONCLUSIONThe blood supply by the middle cerebral artery is similar between the patients receiving obturator prosthesis insertion for maxillary defect and the subjects with full denture.

Adult ; Aged ; Case-Control Studies ; Cerebrovascular Circulation ; Denture, Complete ; Female ; Humans ; Male ; Mastication ; physiology ; Maxilla ; injuries ; Maxillofacial Prosthesis ; Middle Aged ; Middle Cerebral Artery

Objective To observe the expression of STEAP4 gene(a novel obesity-related gene) during the period of human preadipocyte differentiation and to explore the relationship between the STEAP4 gene expression and adipocytes differentiation,adipogenesis.Methods Human preadipocytes were cultured and differentiated into the matured adipocytes in vitro.Adipocytes morphology and lipid accumulation were observed during this process.Total RNA was extracted from adipocytes at various time points (preadipocyte,Day 0,Day 4,Day 6,Day 8,Day 11,Day 14,and Day 17) and the level of STEAP4 mRNA expression was measured by fluorescent real-time quantitative reverse transcriptase-polyme-rase chain reaction(RT-PCR).Results The level of STEAP4 mRNA expression remained high in preadipocytes.In the presence of differentiation medium (Day 4),there was a transient upregulation in the expression of STEAP4 gene.After that,with the human preadipocytes being differentiated into matured adipocytes,the expression of STEAP4 mRNA was downregulated and reached the lowest level in fully differentiated adipocytes.There was a significant difference between any 2 detected phases in the level of STEAP4 mRNA expression (Pa

Objective To analyze the surveillance results and grasp the situation of Keshan disease in Wudalianchi city Heilongjiang province.Methods In 2009,Kaifa village was selected as the surveillance point in Wudalianchi city,total resident population were monitored by routine clinical examination and 12-lead electrocardiogram(ECG) tracing.Suspected cases with Keshan disease were taken chest X-ray,and Keshan disease was diagnosed based on Keshan Disease Diagnostic Criteria (WS/T 210-2011).Results A total of 795 people were investigated,including 397 males and 398 females.Eighteen people were found to be the patients with Keshan disease,of which 13 cases were latent Keshan patients,5 cases were chronic Keshan patients.The overall detection rate was 2.27％,aged 24 to 83 years old.There was no acute type and subacute type of Keshan disease in the surveillance point.Twenty nine cases of abnormal ECG were detected,the detection rate was 3.65％ (29/795),of which the 18 patients with Keshan disease were all had abnormal ECGs,mainly taken the form of ST-T changes and completely right bundle branch blocked.Six cases of male patients with Keshan disease were detected,the detection rate was 1.52％ (6/397); 12 cases of female patients with Keshan disease were detected,the detection rate was 3.01％ (12/398).Conclusions There is still potential and chronic Keshan disease cases in Wudalianchi city.We must keep on the monitoring on Keshan disease,master the dynamical changes of the disease conditions,and carry out the targeted prevention and control of Keshan disease.

OBJECTIVEThe prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.

METHODSPrimary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.

RESULTSThe Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).

CONCLUSIONSiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.

Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; genetics ; Bone Marrow Cells ; cytology ; metabolism ; Caspase 3 ; metabolism ; Cells, Cultured ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Small Interfering

Overexpression of recombinant Human Cu, Zn-Superoxide Dismutase (rhCu, Zn-SOD) in E. coli results in the form of insoluble inclusion body. Purity of rhSOD inclusion body was over 80% by isolation and purification. After preliminary renaturation by conventional dilution or dialysis, enzyme preparations was respectively purified by using Copper Metals-Chelating Affinity Chromatography (Copper-MCAC). RhSOD specific activity purified by MCAC (from the sample renatured partly by dialysis) was 2.2 times as much as that by dialysis and protein recovery was 64%. RhSOD specific activity purified by MCAC (from the sample renatured partly by dilution) was 5.3 times as much as that by dilution and protein recovery was 25%. The two rhSOD preparations purified by MCAC had specific activities about 5000 u/mg and activity recoveries were all over 130% of the enzyme activities in the samples renatured partly by dilution or dialysis. The above-mentioned results indicated that Copper-MCAC resulted in a purification and further renaturation of target protein. SDS-PAGE showed that the target protein rhSOD (19 kD) was purified homogeneously and NBT activity identification proved that the purified and renatured rhSOD had very strong SOD activity. In conclusion, Copper Metals-Chelaing Affinity Chromatography appears to be a simple, rapid and efficient procedure for purifying and further renaturing rhCu, Zn-SOD by dilution or dialysis. The method provided a new idea for purifying and renaturing recombinant proteins expressed in the form of inclusion body in E. coli.
Chelating Agents ; chemistry ; Chromatography, Affinity ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Inclusion Bodies ; genetics ; Protein Renaturation ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Superoxide Dismutase ; biosynthesis ; genetics

Animals ; Fatty Liver ; metabolism ; pathology ; Glycine ; pharmacology ; Liver ; metabolism ; pathology ; Male ; PPAR gamma ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the incidence of JAK2V617F gene point mutation in patients with myeloproliferatives diseases (MPD) and its clinical significance.

METHODSGenomic DNA from bone marrow and peripheral blood cells were extracted from 68 patients with MPD. Allele specific polymerase chain reaction was used to amplify the exon 12 of JAK2 gene which harbours V617F mutation. The PCR products were identified by DNA sequencing. JAK2V617F gene point mutation and its impact on peripheral blood cells were analyzed.

RESULTSThe incidence of JAK2V617F mutation in 68 patients with MPD was 65.28 %. The positive rate of JAK2V617F point mutation was 77.77 % in patients with PV (36/59), 56.52 % in patients with ET (23/59) and 44.44 % in patients with IMF (4/9). In all groups, the incidence of JAK2V617F point mutation in bone marrow and peripheral blood were equal. Patients with JAK2V617F mutation in PV group had higher counts of white blood cell and hemoglobin in peripheral blood than patients without JAK2V617F point mutation (P <0.05). Patients with JAK2V617F mutation in ET group had higher counts of white blood cell than those without JAK2V617F mutation (P <0.05); there was no significant difference in platelet count.

CONCLUSIONJAK2V617F point mutation can affect the hematologic features, which may be of diagnostic value for MDP with negative BCR-ABL gene.

Adolescent ; Adult ; Aged ; Amino Acid Substitution ; Base Sequence ; Female ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Myeloproliferative Disorders ; enzymology ; genetics ; Point Mutation ; Young Adult