ABSTRACT
To develop a genetic transformation method of Aureobasidium pullulans and T-DNA insertion for high-efficient screening of polymalic acid (PMA) producing strain. Agrobacterium tumefaciens-AGL1, containing the selection genes encoding hygromycin B phosphotase or phosphinothricin acetyltranferase, was used to transform Aureobasidium pullulans CCTCC M2012223 and transformants were confirmed by colony PCR method. Transferred DNA (T-DNA) insertional mutants were cultured in microwell plate, and screened for high-titer PMA producing strain according to the pH response model. DNA walking was used to detect the insertion sites in the mutant. Results show that the selection markers could stably generated in the transformants, and 80 to 120 transformants could be found per 10(7) single cells. A high-titer PMA mutant H27 was obtained, giving a good PMA production caused by the disruption of phosphoglycerate mutase, that increased by 24.5% compared with the control. Agrobacterium tumefaciens-mediated transformation and high-efficient screening method were successfully developed, which will be helpful for genetic transformation of Aureobasidium pullulans and its functional genes discovery.
Subject(s)
Agrobacterium tumefaciens , Ascomycota , Genetics , Metabolism , DNA, Bacterial , Malates , Metabolism , Polymerase Chain Reaction , Polymers , Metabolism , Transformation, GeneticABSTRACT
Objective To investigate the protective effect of somatostatin (SS)combined with growth hormone (GH) in treatment of intestinal mucosal barrier injury in rabbits with severe acute pancreatitis (SAP), as well as its clinical significance. Methods Seventy-two rabbits were equally assigned into model group (SAP group), SS treated group (SS group) and SS combined with GH treated group (SS + GH group). SAP models were induced by retro-injection of 5% sodium taurocholate into the pancreatic duct. After modeling, all rabbits were given 5 % glucose saline daily.The rabbits in SS group and SS+GH group were continuously Given SS (3.5μg·kg-1·h-1)for 48 hours. Besides, the rabbits in SS+GH group were subcutaneously injected with 0.15 IU/kg of GH at the 1st and the 24th hours after modeling. The levels of serum amylase, serum tumor necrosis factor-α (TNF-α) and plasma diamine oxidase were measured at the 6th, 12th, 24th and 48th hours after modeling. The pathological changes of pancreatic tissue and ileal mucosa were observed. Survival rate was calculated. Data were analyzed using SPSS 16.0 software. The univariate analysis was used to compare the difference among groups. Results In SS+GH group, the levels of serum TNF-α and plasma diamine oxidase were (2. 43 ± 0. 14) pg/ml and (4. 61 ± 0. 45) U/L at the 24th hour respectively, and were (2.08±0.23) pg/ml and (3.75±0.47) U/L at the 48th hour, respectively,which were lower than those in SAP group and SS group [(2.80 0.30) pg/ml and (8.74 ± 1.77)U/L, respectively, at the 24th hour; (2. 45±0.12) pg/ml and (5. 02±0.95) U/L, respectively, at the 48th hour)]with significant difference (P＜0.05). The inflammation in pancreas and ileal mucosa was alleviated, and the integrity of bowel mucosa was improved. Survival rate of SS+GH group was significantly higher than SAP group and SS group. There was no significant difference in level of serum amylase between SS+GH group and SS group. Conclusion The combination of SS with GH may enhance the function of intestinal mucosa barrier and improve the prognosis of SAP in rabbits.
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<p><b>BACKGROUND AND OBJECTIVE</b>Transbronchial needle aspiration (TBNA) and endobronchial ultrasound-guided TBNA (EBUS-TBNA) have been applied to the diagnosis for mediastinal lymph nodes. The aim of this study is to evaluate the clinical value and safety of TBNA and EBUS-TBNA on hilar and mediastinal lymph nodes of lung cancer patients.</p><p><b>METHODS</b>Two hundred fifty patients with suspected lung cancer were enrolled. All petients with hilar and/or m lymphoadenopa-ediastinal thy found by CT scan received TBNA, biopsy and brushing. EBUS-TBNA was performed in 15 patients among them.</p><p><b>RESULTS</b>Lung cancer were confirmed in 180 patients by TBNA, biopsy and brushing. The positive rates were 82.86%, 51.24% and 45.45%. Fifteen patients after EBUS-TBNA had a positive rate of 91.67%.</p><p><b>CONCLUSION</b>TBNA and EBUS-TBNA were proved to be safe procedure with a high yield for the diagnosis ofhilar and mediastinal lymph nodes in lung cancer patients.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Biopsy, Fine-Needle , Methods , Bronchi , Diagnostic Imaging , Pathology , Endosonography , Methods , Lung Neoplasms , Diagnosis , Diagnostic Imaging , Pathology , Lymph Nodes , Pathology , Mediastinum , PathologyABSTRACT
Objective To investigate the infectious features of rubella virus (RV) JR 23 strain in central nervous system (CNS). Methods RV JR 23 strain infected human primary cerebral neural cell culture in vitro and Balb/c mice, which were given dexamethasone and cytoxan before infection, via peritoneal injection. Viral pathogenecity was observed postinfection and RV antigens were detected in human cerebral neural cells by IFA and immunohistochemical method. Cerebral tissues were observed by HE staining and ABC methods postinfection.Results JR 23 strain didn't produce cytopathic effect (CPE). The proliferation of JR 23 strain reached highest titer of 10 3TCID 50 /ml at 72 h postinfection and decreased gradually. RV antigens were positive in cerebral neural cells, especially around the nuclei. Focal cytopathic areas were observed in cerebral cortical area and so did neuron necrosis around by gliacytes formed stellitorsis, neuronophagia and glial nodule. RV antigens could be seen in all cerebral area, but most localized in cortical area. Pathological features were basically the same among the infected groups. The infection rates of de xamethasone, cytoxan group and the group without intervention were 60%、90% and 50%, repectively. Conclusion RV JR 23 strain is not cytocidal to human neural cells and the pathological lesions induced by JR 23 strain in CNS of mice are mainly focal or dotted neuron necrosis.
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AIM: To investigate the expression of vascular endothelial growth factor (VEGF) and tumor necrsis factor-alpha (TNF-?) in synovium of rats with adjuvant arthritis (AA) and the relationship between the pathological score and the expression of VEGF and TNF-? protein. METHODS: AA was produced in Wistar rats by inoculating complete Freund's adjuvant (CFA). The arthral pathological score was calculated, production of VEGF and TNF-? protein were assayed by histoimmunochemical staining at different stage after CFA inoculation. RESULTS: In AA group, the pathological score and expression of VEGF protein in synovium increased significantly (P