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<p><b>OBJECTIVE</b>To investigate clinical effects of improved anatomical locking plate internal fixation through tarsal sinus incision in treating Sanders III and IV calcaneal fractures.</p><p><b>METHODS</b>From February 2015 to October 2016, 35 patients with Sanders III and IV calcaneal fractures treated by improved anatomical locking plate internal fixation through tarsal sinus incision were collected, including 22 males and 13 females aged from 22 to 68 years old with an average of (42.3±12.7) years old. According to Sanders classification, 23 patients were type III and 12 patients were type IV. Postoperative complications were observed, Bö hler angle and Gissane angle before and after operation were compared, and Maryland foot function scoring standard was evaluated at 12 months after operation.</p><p><b>RESULTS</b>All patients were followed up from 12 to 20 months with an average of (14.5±2.0) months, the length of incision ranged from 4.0 to 5.5 cm with an average of (4.7±0.4) cm. Superficial infection occurred in 1 patient, delayed union in 2 patients, gastrocnemius nerve injury in 1 patient. No complications such as steel plate exposure and fracture reduction loss occurred. Wound healing time ranged from 14 to 28 days with an average of(15.4±4.7) days, the fracture healing time ranged from 8 to 14 weeks with an average of (9.8±1.9) weeks. Bö hler angle increased from preoperative (9.81±14.28)° to 3 days after operation (26.35±11.04)°, and (25.96±10.79)° at 12 months after operation(<0.05). Gissane angle ranged from preoperative (122.54±16.79)° to 3 days after operation (120.85±11.88)°, and (120.62±11.44)° at 12 months after operation and had statistical meaning. Maryland score increased from 12.66±4.10 before operation to 92.20 ±7.82 at 12 months after operation, and 32 patients got excellent results, 2 good and 1 moderate.</p><p><b>CONCLUSIONS</b>Improved anatomical locking plate internal fixation through tarsal sinus incision in treating Sanders III and IV calcaneal fractures, which has advantages of less incision, less soft tissue injury, better fracture reduction and fixation, could receive good reduction and fixation. It is an effective method for Sanders III and IV fracture of calcaneus fracture.</p>
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Tianma(the tuber of Gastrodia eleta) is a widely used and pricy Chinese herb. Its counterfeits are often found in herbal markets, which are the plant materials with similar macroscopic characteristics of Tianma. Moreover, the prices of Winter Tianma(cultivated Tianma) and Spring Tianma(mostly wild Tianma) have significant difference. However, it is difficult to identify the true or false, good or bad quality of Tianma samples. Thus, a total of 48 Tianma samples with different characteristics(including Winter Tianma, Spring Tianma, slice, powder, etc.) and 9 plant species 10 samples of Tianma counterfeits were collected and analyzed by HPLC-DAD-MS techniques. After optimizing the procedure of sample preparation, chromatographic and mass-spectral conditions, the HPLC chromatograms of all those samples were collected and compared. The similarities and Fisher discriminant analysis were further conducted between the HPLC chromatograms of Tianma and counterfeit, Winter Tianma and Spring Tianma. The results showed the HPLC chromatograms of 48 Tianma samples were similar at the correlation coefficient more than 0.848(n=48). Their mean chromatogram was simulated and used as Tianma HPLC fingerprint. There were 11 common peaks on the HPLC chromatograms of Tianma, in which 6 main peaks were chosen as characteristic peaks and identified as gastrodin, p-hydroxybenzyl alcohol, parishin A, parishin B, parishin C, parishin E, respectively by comparison of the retention time, UV and MS data with those of standard chemical compounds. All the six chemical compounds are bioactive in Tianma. However, the HPLC chromatograms of the 10 counterfeit samples were significantly different from Tianma fingerprint. The correlation coefficients between HPLC fingerprints of Tianma with the HPLC chromatograms of counterfeits were less than 0.042 and the characteristic peaks were not observed on the HPLC chromatograms of these counterfeit samples. It indicated the true or false Tianma can be identified by either the similarity or characteristic peaks on HPLC fingerprint. Comparing the Winter Tianma with Spring Tianma showed that the HPLC chromatograms of 15 winter Tianma samples and 11 spring Tianma samples were similar at the mean correlation coefficient of 0.908. But the intensity of the characteristic peaks were different between the two groups of Tianma samples, i.e. the intensity of gastrodin, paishin A and C in winter Tianma was lower than those in spring Tianma. The Winter Tianma and Spring Tianma could be discriminated by either the Fisher unstandardized discrimination function or Linear discriminant function, based on the peak areas of 11 common peaks on HPLC chromatograms as variate.
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Glioma is the most common form of brain cancer. Despite recent advances in the treatment of solid tumors, there are few effective treatments for malignant gliomas due to its infiltrative nature. It has important significance to improve the treatment of glioma through in-depth understanding the intracerebral metabolic characteristics and pharmacokinetics of chemotherapeutics. Brain microdialysis (B-MD), an effective method to monitor central nervous system anticancer drug disposition, conditions of drugs through the blood-brain barrier, basic pathophysiologic metabolism, bioactive compounds and the changes of neurotransmitter in brain, provides the unique opportunity to allow the simultaneous determination of unbound concentrations of drugs in several tissues, and directly measure gliomas biochemistry continuously. B-MD has been able to monitor the change of brain drugs, metabolites and neurotransmitters, dynamic analysis of the drug concentration and pharmacological effect after administration, pharmacodynamic interaction between drugs, receptor mechanism of drug transport, as well as feedback information of internal environment. B-MD is expected to provide reference for clinical individual chemotherapy of glioma, but also provide powerful tools for the evaluation of new anticancer drugs in vivo. In this review, a comprehensive overview of B-MD for studies on glioma is elucidated with special emphasis on its application to neurochemistry and pharmacokinetic studies.
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Animales , Humanos , Antineoplásicos , Farmacocinética , Barrera Hematoencefálica , Neoplasias Encefálicas , Metabolismo , Glioma , Metabolismo , Espectroscopía de Resonancia Magnética , Metabolómica , Métodos , Microdiálisis , Métodos , Neurotransmisores , Farmacocinética , Preparaciones Farmacéuticas , Metabolismo , Tomografía de Emisión de PositronesRESUMEN
<p><b>OBJECTIVE</b>To investigate the expression of metabotropic glutamate receptor 4 (mGluR4) in cardiomyocytes differentiated from mouse embryonic stem cells (ES cells).</p><p><b>METHODS</b>ES cells were differentiated into cardiomyocytes with hanging-drop cultures. Retinoic acid (RA) and dimethyl sulfoxide (DMSO) were used as positive and negative controls, respectively. The co-expression of cardiac sarcomeric protein (α-actinin or troponin-T) and mGluR4 were verified by immunocytochemistry and flow cytometry analysis. The mRNA and protein expressions of mGluR4 were verified by RT-PCR and Western blot analysis, respectively. Meanwhile, the expression of mGluR4 in prenatal mouse heart was also examined.</p><p><b>RESULTS</b>mGluR4 was expressed in both mouse ES cells and ES cell-derived cardiomyocytes. The level of mGluR4 protein expression decreased during the maturation of the cardiomyocytes. The co-expression rate of mGluR4 and Troponin T in the beating embryoid bodies (EBs) was only (3.00 ±1.00)%. On the other hand, mGluR4 gene and protein expressions showed remarkable down-regulation in the development of mouse fetal heart, which was not detected in mouse adult heart.</p><p><b>CONCLUSION</b>The expression of mGluR4 is down-regulated in the cardiomyocyte differentiation of ES cells. The trend of expression is consistent with that in the prenatal mouse heart development.</p>
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Animales , Ratones , Diferenciación Celular , Fisiología , Línea Celular , Células Madre Embrionarias , Biología Celular , Metabolismo , Ratones Endogámicos ICR , Miocitos Cardíacos , Biología Celular , Metabolismo , ARN Mensajero , Genética , Receptores de Glutamato Metabotrópico , Genética , MetabolismoRESUMEN
Herein, solid lipid nanoparticles (SLN) were proposed as a new drug delivery system for adefovir dipivoxil (ADV). The octadecylamine-fluorescein isothiocynate (ODA-FITC) was synthesized and used as a fluorescence maker to be incorporated into SLN to investigate the time-dependent cellular uptake of SLN by HepG2.2.15. The SLN of monostearin with ODA-FITC or ADV were prepared by solvent diffusion method in an aqueous system. About 15 wt% drug entrapment efficiency (EE) and 3 wt% drug loading (DL) could be reached in SLN loading ADV. Comparing with free ADV, the inhibitory effects of ADV loaded in SLN on hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B virus (HBV) DNA levels in vitro were significantly enhanced.
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Humanos , Adenina , Farmacocinética , Aminas , Antivirales , Farmacocinética , Línea Celular , Sistemas de Liberación de Medicamentos , Fluoresceína-5-Isotiocianato , Colorantes Fluorescentes , Glicéridos , Virus de la Hepatitis B , Nanopartículas , Nanotecnología , Organofosfonatos , FarmacocinéticaRESUMEN
Tomatoes ( Lycopersicon esculentum Mill.) are the principal dietary source of Lycopene which is one of carotenoid and is highly beneficial in preventing some diseases such as the cancer and the heart disease. Suppressing the expression of Lcy gene, the main gene regulating the transformation of the lycopene, is a convenient and effective way to enhance the content of lycopene. The primers were designed according to the gene sequence(U46919)and (X86452) in GenBank. The fruit-specific promoter--phytoene desaturase gene(Pds) promoter and the DNA segment of the Lcy gene were isolated from the genome DNA of tomatoes. The 3'end of Lcy DNA segment was connected together by an intron to inform the RNA interferential segment then which was inserted in the expression vector with the Pds promoter to inform the fruit-specific expression vector. The vector was transformed into the tomatoes through the Agrobacterium tumefaciens. Five transformants were obtained. And the PCR proved that the extra-gene was integrated into the tomato genome. The lycopene in the transgenic tomatoes fruit was increased significantly through analysing the contents of lycopene. These results show that regutating biosynthetic enzyme in carotenoid pathway by RNAi can improve the lycopene content of plant-derived products.
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Agrobacterium tumefaciens , Genética , Carotenoides , Metabolismo , ADN de Plantas , Genética , Frutas , Genética , Metabolismo , Liasas Intramoleculares , Genética , Metabolismo , Solanum lycopersicum , Genética , Metabolismo , Oxidorreductasas , Genética , Proteínas de Plantas , Genética , Metabolismo , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , Genética , Interferencia de ARN , Transformación GenéticaRESUMEN
<p><b>OBJECTIVE</b>The prevalence of non-alcoholic fatty liver disease (NAFLD) has markedly increased. Insulin resistance has been implicated in the pathogenesis of NAFLD. This study was aimed at observing the relationship between insulin resistance and NAFLD, and evaluating the role of pioglitazone (PGZ) acting as insulin-sensitizing agents in the prevention and treatment of rat fatty liver induced by high fat feeding.</p><p><b>METHODS</b>The rats were separated randomly into 6 groups: model group I were fed high fat diet for 8 weeks, PGZ prevention group were given PGZ 4 mg/(kg.d) simultaneously, while control group I were fed normal food for 8 weeks; model group II were fed high fat diet for 16 weeks, PGZ treatment group were given PGZ 4 mg/(kg.d) orally simultaneous with high fat diet for 8 weeks after high fat feeding for 8 weeks, control group II were fed normal food for 16 weeks. The rats were sacrificed after 8 weeks and 16 weeks respectively. Liver weight, body weight, serum activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), tumor necrosis factor alpha (TNF-alpha), fasting blood glucose (FBG), fasting plasma insulin (FINS), HOMA (homeostasis model assessment) insulin resistance index (HOMA-IR), and the liver histology of rats of all groups were assayed.</p><p><b>RESULTS</b>After 8 weeks, the liver in model group I showed typical steatosis, accompanied with mild to moderate lobular inflammatory cell infiltration, liver indexes and serum levels of ALT, AST, ALP, TNF-alpha were significantly increased (P<0.05) compared with control group I. Whereas, the degree of hepatic injury was attenuated in PGZ prevention group, liver indexes and serum levels of ALT, ALP were significantly decreased (P<0.05) compared with model group I. After 16 weeks, notable steatosis, and lobular inflammation were observed in model group II rat liver, while the degree of hepatic injury was attenuated in the PGZ treatment group. Liver index, serum levels of ALT, AST, ALP, FINS and HOMA-IR were significantly increased (P<0.05) in model group II compared with control group II. Whereas, in PGZ treatment group, serum levels of AST and FINS showed decreasing tendency, liver indexes, serum levels of ALT, ALP, TNF-alpha and HOMA-IR were significantly decreased compared with model group II.</p><p><b>CONCLUSION</b>Insulin resistance plays a role in the pathogenesis of NAFLD in rats. Pioglitazone can attenuate insulin resistance and biochemical and histological injury in high fat-induced fatty liver in rats.</p>
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Animales , Masculino , Ratas , Alanina Transaminasa , Sangre , Fosfatasa Alcalina , Sangre , Aspartato Aminotransferasas , Sangre , Hígado Graso , Quimioterapia , Metabolismo , Patología , Resistencia a la Insulina , Hígado , Patología , Ratas Sprague-Dawley , Tiazolidinedionas , Usos Terapéuticos , Factor de Necrosis Tumoral alfaRESUMEN
Objective To characterize the antibiotic resistance,homology and carbapenemase genotypes of imipenem resistant Acinetobac1ter baumannii isolated from our hospital,and analyze the clonal relatedness of the test strains.Methods Ninety five strains of imipenem resistant A.baumannii were isolated from August 2003 to December 2004 in the First Affiliated Hospital, College of Medicine,Zhejiang University.The MICs of 16 antimicrobial agents against these strains were determined by agar dilution and E-test method.The homology of these isolates was analyzed by pulse-field gel electrophoresis(PFGE).The coding gene of carbapenemases was amplified.PCR products were purified,cloned and sequenced.Plasmid DNA was extracted and purified.Conjugation and Southern blot were performed to locate the position of oxa 23 gene.Results The resistance rates to ampicillin-sulbactam and cefoperazone sulhactam were 67.9% and 30.2%.Polymyxin E had the lowest resistance rate of 17%. The resistance rate to other antimicrobial agents was higher than 90%.The 95 strains,isolated from 10 clinical units,were classified into 6 clones.Clones A and B were predominant clones.All strains produced carbapenemases which were confirmed as OXA 23 by PCR and sequencing analysis.No plasmid was extracted and conjugation was not successful.Southern bolt showed that oxa-23 gene was located on Apal-digested chromosomal segments about 220 kb and 200 kb in Clones A and B,re spectively.Conclusions OXA 23-producing A.baumannii has become one of the most important multi-resistant pathogens in our hospital.Clones A and B have widely spread in our hospital.Oxa-23 gene is located on chromosomal DNA.
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<p><b>OBJECTIVE</b>To explore the estrogenic activity of ethanol extract from adzuki bean Phaseolus angularis and its effect on progesterone receptor (PR) level of human breast cancer MCF-7 cells.</p><p><b>METHOD</b>The modified MCF-7 cell proliferation assay was used to evaluate the estrogenic activity of adzuki bean extract. And its effect on PR mRNA and protein were addressed using RT-PCR measurements and western blotting analysis respectively in which pure estrogen receptor antagonist ICI 182,780 was employed as the tool.</p><p><b>RESULT</b>The estrogenic activity of adzuki bean extract at various concentrations (10-200 microg x mL(-1)), expressed as proliferative effect (PE) relative to that of solvent control, was examined. The results indicated that the extract of adzuki bean was able to induce MCF-7 cell growth with a maximum at 100 microg x mL(-1). The results of RT-PCR and western blotting showed that PR mRNA and protein could be significantly induced by adzuki bean extract in MCF-7 cells. Moreover, the specific estrogen receptor antagonist ICI 182,780 could block these reactions.</p><p><b>CONCLUSION</b>Ethanol extract of adzuki bean has the estrogen-like activities through the estrogen response pathway.</p>
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Femenino , Humanos , Neoplasias de la Mama , Metabolismo , Patología , Línea Celular Tumoral , Proliferación Celular , Medicamentos Herbarios Chinos , Farmacología , Estradiol , Farmacología , Antagonistas de Estrógenos , Farmacología , Phaseolus , Química , Fitoestrógenos , Farmacología , Plantas Medicinales , Química , ARN Mensajero , Genética , Receptores de Progesterona , Genética , Semillas , QuímicaRESUMEN
<p><b>OBJECTIVE</b>To reproduce an experimental model of alcoholic liver disease in rats and to investigate the preventive and treatment effects of tea polyphenols on alcoholic liver disease.</p><p><b>METHODS</b>68 male Sprague-Dawley rats were randomly divided into 3 groups: alcohol group (gastrically infused with 56% of ethanol once a day with a dose of 7 g/kg body weight for 4, 12 and 24 weeks), tea polyphenols group (gastric infusion with alcohol same as in the alcohol group and with tea polyphenols at 0.25 g/kg bw) and control group (gastric infusion with normal saline). At the end of 4, 12 and 24 weeks, blood samples were collected and then the rats were sacrificed. Liver samples were obtained for routine histological examination and the degree of hepatic steatosis and alcoholic hepatitis were examined. Blood specimens were used for evaluation of alanine transaminase (ALT) and aspartate aminotransferase (AST).</p><p><b>RESULTS</b>(1) The levels of the two transaminases were elevated with the increase of the duration of ethanol feeding and the difference is significant. TP significantly mitigated the increase of ALT and AST activities induced by the alcohol. (2) Histological changes of the liver injury indicated that piecemeal or focal necrosis of hepatocytes was present in the centrilobular area. As fibrosis advanced, broader septa were formed with central-central and centra-portal bridging necrosis. In the TP infusion group, the severity of the pathological changes was significantly milder.</p><p><b>CONCLUSION</b>The results of this study revealed that TP mitigated the development of alcoholic liver disease, and TP may be a potential drug for treatment of alcoholic liver disease.</p>
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Animales , Masculino , Ratas , Flavonoides , Usos Terapéuticos , Hepatopatías Alcohólicas , Quimioterapia , Fenoles , Usos Terapéuticos , Fitoterapia , Polifenoles , Distribución Aleatoria , Ratas Sprague-Dawley , Té , QuímicaRESUMEN
<p><b>OBJECTIVE</b>To evaluate the effect of tea polyphenol(TP) on the rat with alcoholic liver damage.</p><p><b>METHOD</b>Rats were divided into 3 groups, in which 2 groups were stomach perfused with alcohol to result in ALD, and 1 group of them stomach perfused with TP simultaneously. Another group was normal control groups (stomach perfused with drinking water). In the end of 12 weeks, the liver specimen of each rat was observed by anglicizing its tissue damage, and all data collected was performed by statistical analysis in quantum and semi-quantum. Meanwhile cytokines gene express of each group is determined.</p><p><b>RESULT</b>In the end of 12 weeks, alcoholic hepatitis appeared in rat liver. Hepatic injury in alcohol group and TP group were found, but could not be found in normal group. Compared with pure alcohol group, alcoholic liver damage mainly showing with steatosis in TP group were slight, in addition showing liver cellular swelling with small area, with less spot and focal necrosis, none bridging necrosis. Steatosis were slight relatively, mega-bubble steatosis were less found. Collagen deposition of TP group were less than those of pure alcohol group. Gene expression of. cytokine have diversity statistically such as IL-3, IL-4, IL-1R2, IL-6R, IL-7R2, IL-3Ra, IL-R1, IL-13, IL-1R1, IL-7R2, EPO-R, LIFR, IL-1R2, IL-5R2, CSF1, CD27, IL-6R.</p><p><b>CONCLUSION</b>TP is able to attenuate alcoholic liver damage. It's mechanism is possibly due to modulating cytokines gene expression of cytokine.</p>
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Animales , Ratas , Flavonoides , Farmacología , Expresión Génica , Interleucinas , Genética , Hígado , Metabolismo , Patología , Hepatopatías Alcohólicas , Genética , Metabolismo , Patología , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenoles , Farmacología , Plantas Medicinales , Química , Polifenoles , ARN Mensajero , Genética , Distribución Aleatoria , Ratas Sprague-Dawley , Receptores de Interleucina , Genética , Té , QuímicaRESUMEN
<p><b>OBJECTIVE</b>To study the characteristics of the SiO2/R2O3 in non-zonal purple soil in the light of the difference between the high yield and low yield of Salvia miltiorrhiza.</p><p><b>METHOD</b>Both the high yield and low yield drugs developed from the same parental material (k1cy) were chosen in the ecology area in Zhongjiang county, Sichuan Province. By using the air-dried drugs as sample, quadrat sampling method was used to calculat the production. Gravimetric method, volumetric method, and colorimetric method and neutralization titration were used to measure the content of adhesive particle colloids element in the soil. And T-test was used for the variable-difference analysis.</p><p><b>RESULT</b>There was a sharp contrast between the production of high yield and low yield drugs (P < 0.001). The characteristic value of the high yield soil embryology was much higher than that of the low yield soil.</p><p><b>CONCLUSION</b>The characteristic value of the purple soil embryology is one of the important factors of the soil adaptability of the drug and its quality. The weathering degree of the soil is closely related to the yield of the drug.</p>
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Aluminio , Óxido de Aluminio , Ecosistema , Compuestos Férricos , Hierro , Plantas Medicinales , Salvia miltiorrhiza , Silicio , Dióxido de Silicio , Suelo , TitanioRESUMEN
<p><b>OBJECTIVE</b>To evaluate the indications and surgical procedure of bronchial and pulmonary artery sleeve resection for patients with centrally located non-small cell lung cancer, and how to prevent complications.</p><p><b>METHODS</b>From July 1989 to Aug 2000, 32 cases of central NSCLC were treated with bronchial and pulmonary arterial sleeve resection and reconstruction. The results were retrospectively analyzed.</p><p><b>RESULTS</b>The complication rate was 25.0% (8/32), the mortality rate in 30-day postoperation was 6.3% (2/32), the overall 1-, 3- and 5-year survival rate was 82.8% (24/29), 50.0% (11/22) and 33.3% (4/12), respectively.</p><p><b>CONCLUSION</b>Bronchial and pulmonary arterial sleeve resection and reconstruction in the treatment of patients with central NSCLC can not only maximize preservation of functional pulmonary parenchyma and improve patients, quality of life, but also provide an opportunity for those patients with poor pulmonary function to receive surgical resection of the tumor.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Bronquios , Cirugía General , Carcinoma de Pulmón de Células no Pequeñas , Cirugía General , Neoplasias Pulmonares , Cirugía General , Complicaciones Posoperatorias , Arteria Pulmonar , Cirugía General , Procedimientos de Cirugía PlásticaRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of tea polyphenol (TP)'s on liver fibrosis in rats and its possible mechanism.</p><p><b>METHOD</b>Rats were divided into 5 groups, 4 groups of which were stomach perfused with alcohol resulting in alcoholic liver fibrosis, and 3 groups of which were stomach perfused with TP simultaneously. Another group was normal control one. Histological change of rat liver was investigated and quantitative analysis was made in 24 weeks, and rat liver anti-oxidation index and serum endotoxin were determined at the same time.</p><p><b>RESULT</b>Liver fibrosis in TP group was slight, and anti-oxidize index and endotoxin level were markedly improved in comparison with those of alcohol groups.</p><p><b>CONCLUSION</b>Tea polyphenol can protect hepatocytes from fibrosis. Its mechanism is possibly due to cleaning up overfull lipid per-oxidation and reducing the level of endotoxin.</p>
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Animales , Femenino , Masculino , Ratas , Colágeno , Metabolismo , Endotoxinas , Sangre , Flavonoides , Farmacología , Glutatión , Sangre , Hígado , Metabolismo , Patología , Cirrosis Hepática , Sangre , Metabolismo , Hepatopatías Alcohólicas , Malondialdehído , Sangre , Fenoles , Farmacología , Polifenoles , Sustancias Protectoras , Farmacología , Ratas Sprague-Dawley , Superóxido Dismutasa , Sangre , Té , QuímicaRESUMEN
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.