ObjectiveTo investigate the etiology, exacerbating factors and outcome of geriatric patients with acute renal failure (ARF) in emergency and critical internal medicine. MethodsClinical features of elderly inpatients with ARF who were diagnosed in internal medicine of our hospital during the past 10 years were retrospectively analyzed. Patients were divided into community-acquired ARF (CA-AFR) group and hospital-acquired ARF (HA-ARF) group. Comparisons between CA-ARF group and HA-ARF group, and between elderly patients and non-elderly patients with ARF in internal medicine were performed. ResultsAmong 381 elderly ARF patients in internal medicine, there were 218 (57.2%) CA-ARF patients and 163 (42.8%) HA-ARF patients. Most of the HA-ARF patients (153 cases) were distributed in medical intensive care. Compared with CA-ARF group, patients in HA-ARF group were older, with more underlying diseases, a higher ratio of infections and/or heart failure and more severe acute renal failure. Infection and heart failure/severe myocardial isehemia were the major exacerbating factors for HA-ARF. Of the 163 patients with HA-ARF, 146 patients died and 17 patients survived. Patients in the death group had more underlying diseases, a higher ratio of severe infections and heart failure/severe myocardial ischemia, a higher ratio of cases with multiple organ dysfunction syndrome(MODS) and a higher score of APACHE Ⅱ.When compared with non-elderly patients with ARF, the elderly patients had a higher proportion ofHA-ARF, a higher ratio of MODS, a higher score of APACHEⅡ and a higher mortality of ARF.ConclusionsThe elderly critical patients are susceptible to HA-ARF with more chronic underlying diseases. Infection and heart failure/severe myocardial ischemia are the major exacerbating factors.The prognosis depends on the treatment of predisposing diseases and removal of exacerbating factors.

Objective To investigate the methods of prevention and treatment of presacral venous plexus bleeding in pelvic operation.Methods The clinical data of 8 cases of presacral venous plexus bleeding in pelvic operation from 1998 to 2013 were analyzed.Results All 8 cases succeeded in controlling bleeding,The amount of bleeding was 1 000-4 000 mL,the average amount is 2 600 mL.Conclusions The key to prevention of presacral venous plexus bleeding was thorough familiar with pelvic anatomy and a dexterous technique of careful dissection.Massive hemorrhage occurred direct electric coagulation hemostasis was available,the method was simple.

Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals ; Base Sequence ; Cloning, Organism ; Electrophoresis ; Endonucleases ; genetics ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; Gene Targeting ; methods ; Goats ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Zinc Fingers

Objective To establish a doxycycline-controlled immortalized pre-cartilaginons stem cells (IPCSCs) strains, clone parathroid hormone-related peptide[PTHrP(1-36)] gene and construct re- sponsive plasmid, pTRE-PTHrP (1-36). Methods Plasmid pTet-on was transfected into IPCSCs by using LipoinfectaminTM 2000 and then the stable clones were obtained by G418 screening. The doxycyc- line was added into the medium of monoclonal cells that were transiently transfected with plasmid pTRE- 2Hyg-Lue. The total RNA was extracted from PCSCs and the PTHrP(1-36) gene obtained by RT-PCR method. Then, the PTHrP (1-36) gene was subcloned to plasmids of Tet-responsive element with the se- lection marker of hygromycin pTRE-2Hyg to construct recombinant eukaryotic expression plasmid pTRE- PTHrP(1-36). After transferred into E. coli-DH5α, the clone was amplified, the recombinant plasm0ids were purified and identified by double-enzyme digestion. Results The doxycycline induced IPCSCs line was obtained, with 50 times higher than the non-induced cell line. Double enzyme digestion analysis and sequencing showed that the target gene was cloned into recombinant plasmid. Conclusions The induced IPCSCs line can be used to highly express alien genes. The responsive plasmid containing PTHrP (1-36) gene may be premising for rigorous control of PTHrp (1-36) gene expression.

Objective To investigate whether human p27kip1 gene transfection mediated by nanoparticles( NP) can express p27kip1 protein and inhibit the neointimal formation in rat vein grafting models. Method Autogenous vein graft model was established in 120 Wistar rats by transplanting internal branch of jugular vein to carotid artery. Rats were divided into three groups: (1) p27 group; p27kip1 gene mediated by NP were transfected into the veins before anastomosis; (2) control group; the veins were transfected by simple NP; (3) grafting group;the veins were grafted without transfection. The grafted veins were harvested at 3 d, 7 d, 14 d and 28 d respectively after the operation. The exogenous p27 kip1 protein expression in veins was determined by Western blot. Intimal hyperplasia (IH) and PCNA were observed by pathology and analyzed by computer digitizing system. The presence of apoptotic VSMC was demonstrated by TUNEL method. Result p27kip1 gene transfection mediated by NP complex increased protein expression of p27kip1 gene and there was a significant decrease in the intimal average thickness at 7 d, 14 d and 28 d in p27 group (P

Objective To screen a human single-chain variable fragments(scFv)against antiGBM antibody.Methods Using phage display technique,the phage antibody library was panned by antiglomerular basement membrane(GBM)antibody which was coated in a micro-titer plate,one clone was found to have high affinity to anti-GBM antibody.The DNA sequence of the positive clone was determined.Results Along with the increase of rounds anti-GBM antibody specific phage antibody was highly enriched and screening efficiency was increased 137 folds than the firest round.ELISA and competition inhibition assay showed that the scFv had a specific combination character with anti-GBM antibody.DNA sequencing confirmed that the whole gene of scFv was 750 bp,and in accordance with humanized single-chain variable region antibody sequence structure.Conclusion The results suggested that the scFv fragment to anti-GBM antibody could be successfully selected by recombinant phage antibody technique,which will laid an experimental foundation for further research of the therapy of Goodpasture syndrome.

Objective Hypoalbuminemia is a typical symptom of nephrotic syndrome ( NS) , which may result from the loss of much protein with urine.Hyperbilirubinemia is also a common symptom in patients with NS.This study is intended to reveal the relationship between hypoalbuminemia and hyperbilirubinemia in patients with NS by investigating urine bilirubin, albumin( ALB ) and 24-hour urine protein in the NS group, chronic glomerulonephritis ( CGN ) group and postoperative gastroparesis syndrome ( PGS) group ( ALB<35 g/L) .Methods Totally 187 patients with NS, 70 patients with CGN and 64 patients with PGS ( ALB <35 g/L ) were recruited before ALB, urinary protein ( UPR ) , urinary microalbuminuria/creatinine(Umalb/cr) and total bilirubin(TBIL) were detected.SPSS 17.0 Software was used to analyze the difference between the three groups and to reveal the correlations between TBIL and UPR, ALB.Results TBIL, ALB, UPR and Umalb/Cr levels were significantly different between NS, CGN and PGS groups ( one-way ANOVA test, P <0.05), and TBIL was positively correlated with ALB but negatively correlated UPR and Umalb/Cr in both NS and CGN groups (Spearman′s rho test,P<0.05);but no correlation was found between these items in PGS group (Spearman′s rho test,P>0.05) .Conclusion Serum bilirubin of patients with NS is at a low level and shows significant correlations with serum albumin and urinary protein levels.No similar association is found with the other two groups.The results in this study show that the causes of low serum bilirubin in patients with NS may related to the large amount of protein lost in urine.

AIM:Atherosclerotic calcification is highly linked with plaque instability and cardiovascular events .Adenosine monophosphate-activated protein kinase ( AMPK) has been involved in the pathogenesis of various cardiovascular disease .The contributions of AMPKαsubunits to the development of atherosclerotic calcification in vivo remained unknown .We hypothesized that AMPKαsubunits may play a role in the development of atherosclerotic calcification .METHODS: Atherosclerotic calcification was generated by 24-week fed of western diet in ApoE-/-background mice .Calcification was evaluated in aortic roots and innominate arteries of ApoE-/-mice or in mice with dual deficiencies of ApoE and AMPKαsubunits globally ( AMPKα1 and AMPKα2 ) , or vascular smooth muscle cell ( VSMC)-specific or macrophage-specific knockout of AMPKα1 with atherosclerotic calcification pone diet . The mechanism of AMPKα1 in regulating Runx2 was further explored in human aortic VSMC .RESULTS: Ablation of AMPKα1 but not AMPKα2 in ApoE-/-background promoted atherosclerotic calcification with increased Runt -related transcription factor ( Runx2 ) expression in VSMC compared with ApoE-/-mice.Conversely, chronic administration of metformin, which activated AMPK, markedly reduced ath-erosclerotic calcification and Runx2 expression in ApoE-/-mice but had less effects in ApoE-/-/AMPKα1 -/-mice.Furthermore, VSMC-but not macrophage-specific deficiency of AMPKα1 in ApoE-/-background promoted atherosclerotic calcification in vivo com-pared with the controls .AMPKα1 silencing in human aortic VSMC prevented Runx 2 from proteasome degradation to trigger osteoblastic differentiation of VSMC .Conversely , activation of AMPK led to Runx 2 instability by inducing its small ubiquitin-like modifier modifi-cation (SUMOylation).Protein inhibitor of activated STAT-1 (PIAS1), the SUMO E3-ligase of Runx2, was directly phosphorylated by
AMPKα1 at serine 510, to enhance its SUMO E3-ligase activity.Ablation of PIAS1 serine 510 phosphorylation inhibited metformin-in-duced Runx2 SUMOylation, and subsequently prevented the effect of metformin on reducing oxLDL-triggered Runx2 expression in hu-man aortic VSMC.CONCLUSION:Deficiency of AMPKα1 in VSMC increases Runx2 expression and promotes atherosclerotic calcifi-cation in vivo.AMPKα1 phosphorylates PIAS1 to enhance Runx2 SUMOyalation and subsequent degradation .

Objective To evaluate the effect of magnetic resonance thoracic ductography (MRTD)in the diagnosis of right thoracic duct.Methods MRTD data were analyzed retrospective,and the detection rate of right thoracic duct was summarized and compared with that of lymphoscintigraphy,direct lymphangiography and operation.Results 12 cases of right thoracic duct were detected in 1547 cases of MRTD.The detection rate was 0.78%,in which 1 case was complied with total internal organs inversion,and 1 case with right aortic arch.Lymphoscintigraphy were performed in all 12 cases and right thoracic duct were detected in 4 cases.Direct lymphangiography were performed in 4 cases and right thoracic duct were observed in all of them.7 cases of them received right lum-bar duct adhesiolysis.Conclusion MRTD is a noninvasive method for diagnosis of right thoracic duct,which providing useful guid-ance for surgical operation.Its detection rate and diagnostic accuracy are higher than those of lymphoscintigraphy.

Objective To develop whole breast irradiation with volumetric-modulated arc therapy (VMAT)-based hybrid intensity-modulated radiotherapy (IMRT) after breast conserving surgery for breast cancer,and to evaluate its value in clinical application.Methods Ten patients with breast cancer undergoing breast conserving surgery were enrolled.Two radiotherapy plans were designed based on hybrid fixed-beam IMRT/IMRT technique and hybrid VMAT/IMRT technique,respectively.One group received whole breast irradiation with a prescribed dose of 50 Gy in 25 fractions.The other group received whole breast irradiation with a prescribed dose of 50 Gy in 25 fractions,as well as simultaneous integrated boost to the tumor bed with 60 Gy in 25 fractions.The dosimetric parameters and delivery efficiency were compared between the two plans using paired t test.Results For patients treated with whole breast irradiation alone,there were no significant differences in conformity index and homogeneity index of target volume between the two plans (P=0.866,0.056);however,compared with the hybrid fixed-beam IMRT/IMRT technique,the hybrid VMAT/IMRT technique significantly increased the doses to organs at risk (OARs) and the number of monitor units (MUs) in the intensity-modulated field (P=0.000-0.050,P=0.002).For patients treated with whole breast irradiation with a simultaneous integrated boost to the tumor bed;however,the hybrid VMAT/IMRT technique significantly reduced the doses to the lung and spinal cord,number of MUs in intensity-modulated field,and delivery time compared with the hybrid fixed-beam IMRT/IMRT technique (P=0.004,0.001,0.000,0.000).Conclusions For patients treated with whole breast irradiation with a simultaneous integrated boost to the tumor bed,the hybrid VMAT/IMRT technique is highly promising for clinical application due to better OAR sparing and higher delivery efficiency.