Bioimprinting is a new developed technique to improve the characteristics of enzymes.Bioimprinting by lauric acid was conducted to improve the esterification activity of lipase PS in sol-gel immobilization process with methyltrimethoxysila(MTMS) and tetramethoxysila(TMOS) as the precursors.Results generated by checking the esterification activity and scanning electron microscope showed that bioimprinting can enhance the specific activity and thermal stability of lipase PS.The bioimprinting system was optimized by orthogonal experiment,and the optimal condition for lipase bioimprinting is water/silane molar ration(R) 12,polyethylene glycol(PEG) 120?l,and lauric acid 0.15 mmol.Compared with the free enzyme and the non-imprinted enzymes,the specific activity of imprinted enzymes has been improved 44.3 fold and 2.4 fold,respectively.Imprinted lipase show better thermal stability,and the relative activity is 58% after incubated in 80 ℃ for 0.5 h,while no activity was detected for the free enzyme.

Objective To evaluate the clinical outcome and relative factors of intervention treatment for atherosclerotic renal artery stenosis in elderly patients.Methods The clinical data of 79 patients diagnosed as atherosclerotic renal artery stenosis by angiography and treated by revascularization were analyzed.Results There were 55(69.6%)successes and 24(30.4%)failures in decreasing blood pressure and 28(35.4%)successes and 51(64.6%)failures in improving renal function after intervention treatment.Predictors of favorable outcome of intervention treatment in decreasing blood pressure were related to lower urine protein,higher glomerular filtration rate,higher systolic and diastolic blood pressure before treatment,lower resistance index(RI)of renal artery,and no complication of cerebral vascular diseases.Predictors of favorable outcome of intervention treatment in improving renal function were related with percentage of angiographic stenosis,category of antihypertension and lower urine protein.The logistic regression analysis showed that the percentage of angiographic stenosis was the most important predictor of intervention treatment for blood pressure control,age and level of serum creatinine before intervention treatment were the most important predictors of intervention treatment for improving renal faction.Conclusion Percentage of stenosis(≥85%),age(133 ?mol/L)can be used as the predictors of therapeutic success for renovascular stenosis in older patients.

OBJECTIVE: To establish a rapid and simple gas chromatographic-mass spectric method for qualitative and quantitative analysis of lidocaine in blood and cerebrospinal fluid(CSF). METHODS: Following an acidification of HCl, blood or CSF was alkalinized with NaOH (pH=9) and extracted with ether for two times. Evaporated in a water bath and with an air velocity of nitrogen gas, extract was dissolved with ethanol and analyzed by a gas chromatographic-mass spectrum method, lidocaine was analyzed qualitatively and quantitatively by GC/MS (SIM:86, 58, 72, 87). RESULTS: Linear range of lidocaine detected in blood or CSF by this method is 1.0-60.0 microg x mL(-1) (r=0.9999), the minimum detected concentration of lidocaine was 0.02 microg x mL(-1) (S/N=3), recovery is at 85%-103%. This method was used in the determination of lidocaine in dog model died of the anesthesia with lidocaine. CONCLUSION This study provided a gas chromatographic-mass spectric analysis for lidocaine in blood and CSF. This method was more selective, little interferefering, more sensitivities and simpler. It could be used in the detection of lidocaine in biological fluids.
Anesthesia, Epidural ; Anesthetics, Local/cerebrospinal fluid* ; Animals ; Dogs ; Forensic Medicine ; Gas Chromatography-Mass Spectrometry/methods* ; Humans ; Injections, Intravenous ; Lidocaine/cerebrospinal fluid* ; Sensitivity and Specificity

Based on site-specific transposition of an expression cassette into a baculovirus shuttle vector (Bacmid) which propagated in Escherichia coli, the Bac-to-Bac System provides a rapid and efficient method to generate recombinant baculoviruses and is widely used for high level expression of heterologous proteins. And the efficiency of recombinant baculovirus infecting cells plays an important role on the protein expression. In this study, we introduced an EGFP expression cassette driven by polyhedrin promoter into the p74 locus of Bacmid by homologous recombination. The target Bacmid-egfp was then transformed into E. coli DH10B containing the transposition helper plasmid to gain a new transposition receipt strain E. coli DH10Bac-egfp. Because of the intact attTn7 sites and lacZ', target gene cloned in a pFastBac vector can be transposed into the Bacmid-egfp shutter vector to construct recombinant baculovirus, which would allow the tracing of the target protein expression and the recombinant Bacmid transfection or recombinant baculoviral infection under fluorescence microscopes. Recombinant virus Bac-egfp-DsRed was constructed by transposing DsRed into the Bacmid-egfp in E. coliDHl0Bac-egfp, and the Sf9 cells infected with the recombinant virus expressed DsRed and EGFP efficiently. Another protein IL-6 fused with 6 x his tag was expressed and purified sucessfully from Sf9 cells infected with recombinant virus Bac-egfp-6 x his-IL6 constructed by the improved Bac-to-Bac system.
Animals ; Baculoviridae ; genetics ; Green Fluorescent Proteins ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Spodoptera

The specific inhibition of angiotensin II action at AT(1) receptors by losartan has been shown to decrease peripheral insulin resistance in type 2 diabetic patients and animal models. We examined the effect of losartan on the expression of insulin receptor substrate 1 (IRS-1), protein kinase B (PKB) and glucose transporter 4 (GLUT4), as well as the phosphorylation status of IRS-1 and the association between IRS-1 and phosphatidylinositol (PI) 3-kinase in skeletal muscle from fat-fed and-streptozotocin (STZ)-treated rats, an animal model of type 2 diabetes mellitus. In addition, the effects of losartan on GLUT4 translocation in muscle cells and on insulin sensitivity were also evaluated. Muscle tissues were isolated from male losartan-treated and untreated normal or non-insulin-dependent diabetes mellitus (NIDDM) rats with a dose of 4 mg/kg per day for 6 weeks. Oral administration of losartan improved insulin sensitivity, which was determined by an oral glucose tolerance test (OGTT). In skeletal muscles, the protein levels of IRS-1, PKB and GLUT4 in NIDDM rats were not significantly different from those of the control rats, and they were not affected by losartan. The levels of IRS-1 tyrosine phosphorylation, PI 3-kinase activity associated with IRS-1 and PKB activation after stimulation with insulin in muscle tissue of NIDDM rats were significantly decreased (P<0.01) compared with those in the control rats, while they were not increased by losartan. Losartan had a major effect on GLUT4 translocation in myocytes, as it significantly increased (P<0.05) the insulin-induced amounts of GLUT4 in plasma membrane (PM) and T-tubules (TT) in myocytes from NIDDM rats. Consistent with these results, the plasma glucose level in losartan-treated NIDDM rats was decreased (P<0.05) compared with that in untreated NIDDM rats. Our results suggest that losartan may exert beneficial effects on insulin resistance by increasing the translocation of GLUT4 in muscle tissue, which is probably associated with a non-PI 3-kinase-dependent mechanism.
Animals ; Diabetes Mellitus, Experimental ; blood ; drug therapy ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; physiopathology ; Glucose Transporter Type 4 ; Insulin Receptor Substrate Proteins ; Insulin Resistance ; Losartan ; pharmacology ; therapeutic use ; Male ; Monosaccharide Transport Proteins ; biosynthesis ; genetics ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; Phosphoproteins ; biosynthesis ; genetics ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics ; Proto-Oncogene Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-akt ; Rats ; Rats, Sprague-Dawley

ObjectiveTo introduction of perforator flaps,muscle flaps pedicled by descending branch of lateral circumflex femoral artery,method and their clinical application that multiple soft tissue defects of hand are repaire by muliplefoliated tissue flap only branch lateral circumflex femoral artery.MethodsFifteen patients with multiple soft tissue defects of hand were repaired muliplefoliated tissue flap only pedicled bydescending branch lateral circumflex femoral artery.At first,the anterolateral thigh perforator flap was designed and harvested according to the soft tissue defects of hand, then the descending branch lateral circumflex femoral artery was dissected at the same time the segmented perforator flap,fascia lata flap,rectus femoris muscle flap, vastus lateralis muscle flap, vastus intermedius muscle flap and distal spatium intermusculare flap were harvested in need according to distance among soft tissue defects.The muliplefoliated tissue flap was harvested only pedicled by descending branch lateral circumflex femoral artery, at last muscle flaps and fascia lata flaps were covered by skin graft, so the multiple soft tissue defects of hand were repaired in one time.ResultsNo vascular crisis happened. All skin grafts survived well, the contour of all repaired soft tissue defects was good and protective feeling was recovered by skin grafts of all flaps. All cases were got follow-up and the range was from 6 to 20 months(the average was 8.7 months).Wound of donor site healed well, muscle strength of quadriceps and motion of knee were normal. Three cases were excellent,nine cases were well and 3 cases were good, according to upper extremity function evaluation criteria of Chinese Medical Society for the Surgery of the Hand, the rate of good was 80 percent.ConclusionMultiple soft tissue defects of hand can be repaired by muliplefoliated flap only pedicled by descending branch of lateral circumflex femoral artery. Its advantages included reduction of operation time and treatment, good recovery of hand contour and function. It is a good method to repair multiple soft tissue defects of hand.

Homo sapiens longevity assurance homologue 2 (LASS2) is a novel gene isolated from a human liver cDNA library by our laboratory, and it is a human homologue of the yeast longevity assurance gene LAG1 (Saccharomyces cerevisiae longevity assurance gene). According to our previous results, LASS2 could interact with subunit c of vacuolar type H(+)-ATPase (V-ATPase), and the overexpression of LASS2 could inhibit the cell growth of a human hepatocellular carcinoma (HCC) cell line, SMMC-7721. In order to understand the role of the interaction between LASS2 and V-ATPase in HCC cell growth, we transiently transfected plasmid pCMV-HA2-LASS2 into HCCLM3, a HCC cell line without the significant expression of endogenous LASS2. The pH-sensitive fluorescence probes, BCECF and BCECF-AM, were used to measure the intracellular and extracellular H(+) concentrations of HCCLM3 cells respectively. The effect of LASS2 gene on apoptosis was evaluated with Annexin-V/FITC and propidium iodide (PI) by flow cytometry. Western blot was used to detect cytochrome c (Cyt c) in the cytosol and mitochondria, as well as pro-caspase-3 in cytosol. The results showed that the cell growth of LASS2-transfected HCCLM3 cells was significantly inhibited compared with that of the mock control. LASS2 transfection increased intracellular H(+) concentration of HCCLM3 cells, while decreased extracellular H(+) concentration. Moreover, LASS2 transfection significantly enhanced the apoptosis of HCCLM3 cells. In LASS2-transfected cells, the amounts of Cyt c increased in the cytosol, while decreased in the mitochondria. Meanwhile, the expression of pro-caspase-3 in the cytosolic extracts was decreased. These results implicate that LASS2 gene might increase intracellular H(+) of HCC cells via the interaction with V-ATPase, thereby inducing cell apoptosis through mitochondrial pathway.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Sphingosine N-Acyltransferase ; metabolism ; Transfection ; Tumor Suppressor Proteins ; metabolism ; Vacuolar Proton-Translocating ATPases ; metabolism

OBJECTIVETo observe the therapeutic effect of Yangxue Qingnao granule (YXQNG) in treating chronic cerebrovascular insufficiency (CCI) and its possible mechanism.

METHODSEighty-three patients with CCI were randomly divided into YXQNG and nimodipine (ND) groups, the score of vertigo and the change in cerebral blood velocity before and after treatment were observed. And in the animal experiment, the authors adopted bilateral ligation of cervical carotid communis artery to establish CCI rat models in order to observe the effect of YXQNG and ND on incubation period of vertigo in rats and on memory performance.

RESULTSAfter clinical treatment, the vertigo score of YXQNG group was 2.34, and that of the ND group was 4.18, the comparison between the two groups showed that the difference was significant (P < 0.05). After treatment, the middle cerebral artery mean velocity (MCA Vm) of YXQNG group was 64.78 cm/s, vertebral artery mean velocity (VA Vm) was 29.78 cm/s, while that of ND group was 60.34 cm/s and 23.23 cm/s respectively, the comparison between these two groups showing statistical significance and the difference being obvious (P < 0.05). Experimental study showed that the rats in the model group after 12 weeks learning and memory were markedly lowered, the vertigo incubation period significantly lengthened, and compared with that of the model group, learning and memory of the YXQNG group was markedly improved and vertigo incubation period shortened, with the difference from that of the ND group insignificant, P > 0.05.

CONCLUSIONYXQNG could effectively improve CCI patients' vertigo and other clinical symptoms and increase the cerebral blood flow, lessen the vertigo incubation of model group rats, elevate model group rats' memory performance.

Aged ; Aged, 80 and over ; Animals ; Blood Flow Velocity ; drug effects ; Cerebrovascular Circulation ; drug effects ; Cerebrovascular Disorders ; complications ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Learning ; drug effects ; Male ; Memory ; drug effects ; Middle Aged ; Nimodipine ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Vasodilator Agents ; therapeutic use ; Vertigo ; etiology ; physiopathology

OBJECTIVETo investigate the risk factors in postoperative cognitive dysfunction (POCD) induced by patient self-controlled intravenous analgesia (PCIA).

METHODSA case-control study included 103 patients with POCD(P group), assessed by Mini-Mental-State test and 103 cognitive normal controls (NP group). The cases and controls were matched for age, gender,history of operation and PCIA volume dose. The relationship between POCD and various factors was analyzed by univariate and multivariate analysis. Spss 11.5 of statistical software was used for data analysis.

RESULTSData from univariate analysis showed that the history of cerebral trauma, education level and VAS score had significant differences between P group and NP group. Multivariate analysis conformed that the history of cerebral trauma, VAS score and education level were significantly related to POCD induced by PCIA and their ORs (95% CI) were 4.261 (1.344-13.508), 2.364 (1.209-4.624), 0.312 (0.170-0.573) respectively.

CONCLUSIONPatient's history of cerebral trauma and low VAS score were independent risk factors of POCD induced by PCIA and high education level seemed to be a protective factor.

Aged ; Analgesia, Patient-Controlled ; adverse effects ; Case-Control Studies ; Cognition Disorders ; etiology ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Risk Factors

OBJECTIVETo explore the health-related quality of life (QOL) status of patients with chronic rhinosinusitis and nasal polyps.

METHODSOne hundred and twenty patients with chronic rhinosinusitis and nasal polyps and two hundred individuals passing health examination were enrolled by random and their QOL scores were assessed by using QOL instruments including existing SF-36 questionnaire (Chinese version ) and SNOT-20 questionnaire translated into Chinese, of which clinimetric and psychometric properties were tested.

RESULTSThe feasibility, reliability, validity, and responsibility of Chinese version of SNOT-20 questionnaire all passed the test. By the assessment of SF-36 questionnaire, it revealed that scores of six domains such as physical functioning, role physical, bodily pain, mental health, vitality, and general health from patients with chronic rhinosinusitis and nasal polyps were lower than that of control group except social functioning and role emotional (P < 0.05). It showed by Chinese version of SNOT-20 questionnaire that patients exceeded healthy individuals in the scores of twenty items of three domains including physical problems, functional limitations, and emotional consequences, of which the most five important items affecting health status were respectively need to blow nose, thick nasal discharge, lack of a good night's sleep, dizziness, and post-nasal discharge (P < 0.05).

CONCLUSIONSQOL instruments such as Chinese version of SF-36 and SNOT-20 questionnaires can effectively differentiate the QOL status between patients with chronic rhinosinusitis and nasal polyps and healthy individuals. The negative impact of chronic rhinosinusitis and nasal polyps on patients' QOL includes physical functioning, role physical, bodily pain, mental health, vitality, general health, and emotional consequences. The problems of nasal discharge, sleep, and dizziness should be sufficiently emphasized in clinical treatment.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; China ; epidemiology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Nasal Polyps ; epidemiology ; ethnology ; Quality of Life ; Sinusitis ; epidemiology ; ethnology ; Surveys and Questionnaires