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Volume 27 ， Issue 3 ， 2012
- Abstract：Objective:To reconstruct pEGFP-C2-L539fs/47,a HERG nonsense mutant in eukaryotic expression plasmid,and observe the fusion protein expressed in HEK293 cells（human embryo kidney cells）.Methods:After double digestion of pcDNA3-L539fs/47 and pEGFP-C2-HERG with sbf I and Eco91 I,the small product fragment,from pcDNA3-L539fs/47,was subcloned into the big fragment of pEGFP-C2-HERG under T4 ligase.pEGFP-C2-L539fs/47 was identified by agarose gel electrophoresis and sequencing.pcDNA3-L539fs/47 and pEGFP-C2-L539fs/47 were transiently transfected into HEK293 cells by Lipofect,respectively.The expression of fusion protein in HEK293 cells was detected through immunofluorescence,laser confocal imaging scanning in vivo,Western blot and PCR.Results:Mutation region cDNA fragment（about 1 kb） and target vector fragment（about 7.2 kb） were ligated after purification and gel recovery.Agarose gel electrophoresis and sequencing successfully demonstrated eukaryotic expression plasmid pEGFP-C2-L539fs/47,constructed approximately 8.2 kb,sequencing consistent with template gene.The transfection efficiency of recombinant plasmid by fluorescence microscopy was more than60%.Western blot analysis detected pcDNA3-L539fs/47 expression of the protein size 60 KD,the expression of pEGFP-C2 fusion protein size of approximately 90 KD.The L539fs/47 gene expression in HEK293 cells was significant by PCR analysis.Confocal laser imaging showed that pEGFP-C2-L539fs/47 protein was successfully expressed in cytoplasm and cytomembrane of HEK293 cells.Conclusion:pEGFP-C2-L539fs/47 containing the HERG gene mutant was successfully constructed by double digestion method and expressed fusion protein in HEK293 cells,which laid a foundation for the further study on L539fs/47.0|51|2Updated：2026-03-12
- Abstract：Objective:To investigate whether remifentanil induced cardioprotecting effect is associated with expression of toll-like receptor 4（TLR4）,nuclear factor κB（NF-κB） and serum interleukin-6（IL-6）.Methods:Fifty rabbits were randomly divided into 5 groups（n=10） according to the treatment:sham operation group（group A）,ischemia-reperfusion group（group B）,low-dose remifentanil group（group C）,mediate-dose remifentanil group（group D）,and high-dose remifentanil group（group E）.Myocardial TLR4 mRNA levels,NF-κB protein expression and serum levels of IL-6 were observed in 120 min after reperfusion.Results:The myocardial expressions of TLR4 mRNA,NF-κB protein and IL-6 level in sera of groups B,C,D and E were elevated compared with group A.However,remifentanil significantly reduced the levels of TLR4 mRNA,NF-κB protein expression and serum IL-6 in groups C,D and E compared with group B.There were remarkable differences between the groups（P<0.01）.Conclusion:Intravenous remifentanil has protective effect against rabbit myocardial ischemia/reperfusion injury.This effect may be associated with TLR4,NF-κB expressions on myocytes and serum level of IL-6 in a dose-dependent manner.0|31|2Updated：2026-03-12
- Abstract：Objective:To improve the diagnosis and therapeutic effect of occurrence and development of hyponatremia and disorder of acid-base balance among patients with hepatic encephalopathy（HE） by elucidating the regularity and mechanism,as well as its influence on prognosis.Methods:327 HE patients admitted to our hospital from January 1990 to June 2010 were enrolled.Meanwhile 316 patients hospitalized in the medical department of the same hospital were chosen as the control group.Patients in both groups were given the same methods to measure arterial blood gas parameters（pH value,PaCO2,[HCO3-],TCO2,BE and SaO2）,blood biochemistry（[Na+],[K+],[Cl-]）,liver function,kidney function and blood glucose,serum sodium,and thereupon tocalculate the anion gap（AG） and the potential [HCO3-],and acid-base balance disorder.Results:Among the 327 HE patients,hyponatremia was found in 188 cases（57.4%）,of whom 132 patients died（70.2%）.While among the 316 patients in control group,68 presented with hyponatremia（21.5%）,and 19 died（27.9%）.The incidence and mortality were significantly different between the two groups（P<0.001）.All the 327 patients presented with different degrees of acid-base balance disorder and 178 died（54.4%）,in whom 164（50.2%） belonged to simple acid-base balance disorder and 74（45.1%） died,136（41.6%） were dual acid-base balance disorder and 80（58.8%） died,27（8.2%） were triple acid-base disturbance and 24（88.9%） died.Whereas in the control group only 83 patients（26.2%） were recognized as simple and dual acid-base balance disorder,and 18（21.7%） died.There was higher incidence of acid-base balance disorder and mortality rate in HE group than control one（P<0.001）.Conclusion:Hyponatremia is valuable to judge HE patients’ prognosis.The key parameters in the judgment and evaluation on acid-base balance disorder among HE patients are the change of pH values and serum electrolyte values.When pH value ≤ 7.30 or > 7.55,it generally suggests a poor prognosis.0|12|2Updated：2026-03-12
- Abstract：Objective:To determine the prognostic factors of ruptured hepatocellular carcinoma（HCC） and report the management of patients with spontaneous rupture of HCC in a single center during a 5-year period and to evaluate one-stage hepatectomy.Methods:A series of 4,209 patients with HCC were collected at Eastern Hepatobiliary Surgery Hospital from April 2002 to November 2006,of whom 200 patients（4.8%） with ruptured HCC were studied retrospectively regarding their clinical characteristics and prognostic factors.The one-stage therapeutic approach to manage ruptured HCC consisted of initial management by conservative method,transarterial embolization（TAE） or surgical hepatectomy.Results of various treatments were evaluated and compared in the randomly selected 202 patients with no history of rupture during the same study period.Results:A total of 200 patients with spontaneous rupture of HCC were studied who underwent surgical treatment（n=105）,TAE 33 and conservative treatment（ConT 62）.A multivariate analysis using the Cox hazard regression model（including all the patients;n=200） identified surgical hepatectomy as the only independent factor determining a relatively long survival period（P<0.0001）.On the other hand,in a further analysis of the patients in whom surgical hepatectomy was successfully performed（n=105）,which identified a maximum tumor size exceeding 6 cm as significant determinants of a poor 12-month（P=0.036）,and a multivariate analysis did not identify as any inverse independent factor determining relatively long-term survival,only a maximum tumor size exceeding 6 cm exhibited a tendency toward being a determinant factor（P=0.083）.Conclusion:Considering the high propensity to spontaneous rupture,as long as preoperatively clinical evaluation meet surgery requirements,elective one-stage hepatectomy for patients with ruptured HCC is the first treatment option.Prolonged survival could be achieved in selected patients with hepatic resection,although the survival results were inferior to those of the patients who did not have the complication of rupture.0|16|5Updated：2026-03-12
- Abstract：A female patient at the age of 17 received dilator implant on the right part of her face to prepare skin flap for her facial scar.The skin flap was in good conditions before scar cutting,but showed poor blood circulation after being transferred.Hyperbaric oxygen therapy was given at 0.2 MPa（2ATA） in double pure oxygen cabins for 90 min every day for 10 d.The skin flap showed in sound viability and sound blood circulation after the treatment.Hyperbaric oxygen therapy can be used to treat blood circulation disorder in transferred skin flap.0|16|2Updated：2026-03-12
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