As one of the most sensitive cells of endoplasmic retieulum stress (ERS), pancreatic β-cells have an a-bundance of endoplasmic reticulum. Fatty acids cause apoptosis of β-cells and might contribute to β-cell loss in type 2 diabetes mellitus via the induction of ERS. Glucose is an amplifier of the ERS response to fatty acid, leading to increased β-cell apoptosis. ERS response mediates glucolipotoxicity-induced β-cell apoptosis.

Objective To construct and to identify eukaryotic expression vector expressing Islet-brain 1(IB1) gene.Methods Total RNA was extracted from human insulinoma.IB1 gene was amplified by PCR from human IB1cDNA library.The eukaryotic expression vector encoding IB1 was constructed by inserting the IB1 cDNA into EcoR I/Kpn I sites of the pEGFP-N1 vector with the green fluorescent.The construct was transfected into RINm5F cell line,screened by G418.The phase contrast fluorescence microscope,flow cytometer,and Western blot were used to identify the recombinant plasmid and transfeced cell line.Results The RT-PCR products for IB1(AA1-280)generated from human insulinoma was 840 bp.Sequence analysis proved the same sequence as published in Gen-Bank.Two bands showed that pEGFP-N1 vector encoding IB1 digested by EcoR I or Kpn I.Western blot showed IB1 gene was expressed in RINm5F cells.Conclusion The recombinant prokaryotic expression plasmid pEGFP-N1-IB1 has been successfully constructed.

Adult ; Aged ; Atrial Fibrillation ; etiology ; prevention & control ; Bronchoalveolar Lavage ; adverse effects ; Humans ; Male ; Middle Aged

Objective To investigate the effect of augmenter of 1iver regeneration(ALR)on hepatocytes transplanted intraperitoneally in acute hepatic failure(AHF)rats.Methods AHF SD rats were induced by D-gal.The rats were divided into 5 groups randomly:blank control group(only receiving normal saline),transplantation control group(transplanting 2 X 107 hepatocytes from SD rats),ciclosporin A group(CsA group,receiving hepatocytes and CsA 6 days after transplantation),ALRgroup(receiving hepatocytes plus ALR 6 days after transplantation),ALR control group(only receiving ALR 6 days after transplantation).All drugs and cells were intraperitoneally injected.There was a observation period of 14 days after transplantation.The survival rate of receptors and transplanted hepatocytes was observed.The expression levels of CD4,CD8 and CD68 in abdominal cells,the levels of IL-1β and TNF-a in serum and douche of peritoneal cavity were detected.Results The 2-week SuFvival rate in blank control group,transplantation control group,CsA group,ALR group and ALR control group was 0,46.7%,20%,66.7%and 14.3%respectively.Survival rate in AI.R group was higher than in blank control group(P=0.001).Serum levels of d1-2 TNF-a in ALR group was all lower than in blank and transplantation control groups(P＜(0.01).Serum levels of d1-2 IL-18 in ALR group were lower than in transplantation control group.Ascitic levels of d1-2 TNF-a and IL-1β in transplantation control group were higher than in other groups(P＜0.05,P＜0.01).The serum levels of TNF-a and IL-1β in all survival groups were similar tO normal levels on the 14th day after transplantation.The positive expression rate of d1-2 CD68 in grafts of ALR group was(0.5±0.3)%.The positive expression rate of CD68 in grafts of ALR group was obviously lower than in transplantation control group and CsA groups(P＜0.01).The expression of d1-2 CD68,CD4 and CD8 in ALR group was respectively(1.3±1.2)%,(0.1±0.3)%and(0.2±0.1)%,all obviously lower than in transplantation control group(P＜0.01,P＜0.01,P＜0.05).Assembled conglobate hepatocyte noduses were observed in abdominal cavity in rats receiving hepatocytes transplantation during first to third day after transplantation.The number of survival grafts in ALR group was more than that in transplantation control group and CsA group,and infihrated inflammatory cells were less.On the 14th day posttransplantation.small amounts of normal hepatocytes in grafts were observed only in ALR group.Conclusion After HcT plus ALR intraperitoneally,the survival rate of AHF rats was increased.ALR improved survivorship of transplanted hepatocytes in short term.

Objective To investigate whether catalpol is the active component responsible for the Yin tonic effect of Radix Rehmanniae.Methods Young NH mice were injected with triiodothyronine to produce the hyperthyroidism model,while old mice were used as the model of natural aging.The single point radioligand binding assay was carried out to determine the ?-adrenergic receptor density and M-cholinergic receptor density.The learning ability(short term memory) was determined by the Y-maze avoidance test. Results In the ?-adrenergic receptor experiment,the densities were(15.7?5.2) and(20.9?7.2) fmol/mg protein in normal control group and in T3 control group(P

OBJECTIVETo study measurement methods of acromioclavicular and coracoclavicular ligament injuries,its therapeutic effects and complications during internal fixation operation for the treatment of fresh acromioclavicular joint dislocations of Tossy type III.

METHODSFrom July 2003 to May 2012,127 patients with acromioclavicular joint dislocations of Tossy type III were treated with wire fixation from coracoid process to clavicle or hook-plate fixation. The patients were divided into group A (63 cases) and group B (64 cases) according to whether acromioclavicular ligament and coracoclavicular ligament were repaired or not. In group A (ligaments repaired), there were 39 males and 24 females with an average age of (33.25 +/- 8.46) years old (ranged from 17 to 59 years). And in group B (no ligaments repaired), there were 41 males and 23 females with an average age of (34.10 +/- 7.19) years (ranged from 19 to 57 years). The operation times, intraoperative blood loss, postoperative infections, internal fixation failure, recurrence and other complications, together with therapeutic effects were compared between two groups.

RESULTSThe outcome was analyzed according to Karlsson standard. In group A, 54 patients got an excellent result and 9 good according to Karlsson standard;the average operative time was (55.90 +/- 26.56) min; the average intraoperative bleeding amount was (99.80 +/- 50.30) ml; 1 patient had wire broken without re-dislocation at 16 weeks after operation, 3 patients got wound fat liquefaction and recovered after treatment, 1 patient had pain after shoulder joint motion and pain disappeared after implants were taken out. In group B, 52 patients got an excellent result and 12 good according to Karlsson standard; the average operative time was (49.50 +/- 23.14) min; the average intraoperative bleeding amount was (87.30 +/- 46.41) ml; 2 patients got wound fat liquefaction, and 2 patients had pain after shoulder joint motion. All the patients were followed up, and the duration ranged from 9 to 16 months. All internal steel-wire or hook plate were taken out during 4 to 9 months without acromioclavicular joint re dislocation. There were no significant difference in the average operative time, the average intraoperative blood less, complication recurrence rates of fixation failure, wound fat liquefaction, postoperative infection, acromioclavicular joint re-dislocation, and therapeutic effects between two groups.

CONCLUSIONBoth wire and clavicular hook plate fixation, performed for fresh acromioclavicular joint dislocation with Tossy type III, are simple, effective, less invasive method with less blood loss. In addition, the treatment without ligaments repaired could not increase incidence of complications.

Acromioclavicular Joint ; diagnostic imaging ; injuries ; surgery ; Adolescent ; Adult ; Case-Control Studies ; Clavicle ; Female ; Humans ; Joint Dislocations ; diagnostic imaging ; surgery ; Ligaments ; diagnostic imaging ; injuries ; surgery ; Male ; Middle Aged ; Orthopedic Procedures ; methods ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Antigens, CD34 ; metabolism ; Carcinoma, Signet Ring Cell ; genetics ; metabolism ; pathology ; surgery ; Codon ; Diagnosis, Differential ; Exons ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Gastrointestinal Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Gastrointestinal Stromal Tumors ; genetics ; metabolism ; pathology ; surgery ; Humans ; Melanoma ; metabolism ; pathology ; Middle Aged ; Neurilemmoma ; metabolism ; pathology ; Point Mutation ; Proto-Oncogene Proteins c-kit ; metabolism ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; metabolism

Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

Cis-1,2-dihydroxycyclohexa-3,5-diene (DHCD) can be used as a valuable chiral intermediates for applications in pharmaceuticals, aerospace, electrical and fine chemical industries. By on-line detection of toluene dioxygenase (TDO) activity in whole recombinant Escherichia coli JM109 (pKST11) cells that harbored TDO gene under a tac promoter, effects of IPTG and various benzene addition strategies on bioransformation of benzene to DHCD were investigated. When IPTG was used at the beginning of fermentation, the growth of cells was inhibited and TDO activity only maintained for 4 hours while same experiments with addition of IPTG at 6h or 8h generated TDO activity for 18 hours. Suitable induction time for IPTG was in the cell logarithmic growth phase and 0.5 mmol/L IPTG was sufficient for inducing maximum TDO activities. Benzene strongly inhibited the activity of TDO which catalyses the conversion of benzene to DHCD. It was found that both cell growth and TDO activity was remarkably inhibited by feeding of benzene vapor, only 7.5 g/L DHCD was obtained. While the benzene inhibition effect was ameliorated by two-liquid phase culture fermentation in which liquid paraffin was used as second phase in the broth. Using different initial ratios of paraffin to benzene in fed-batch culture, DHCD contents were increased to 22.6 g/L, which was 3-fold more compared with that in benzene vapor culture. A further improvement of DHCD production was achieved when the mixture of liquid paraffin and benzene was added continuously by peristaltic pump, the DHCD contents were increased to a final concentration of 36.8 g/L. It was proven that the key to improving DHCD production by recombinants is to prolong TDO activity in cells, which can be achieved by using suitable addition benzene strategies.
Benzene ; metabolism ; Cyclohexanols ; metabolism ; Escherichia coli ; genetics ; metabolism ; Fermentation ; physiology ; Oxygenases ; genetics ; metabolism