Objective To investigate the interference effects of orexin A on cell proliferation of the insulin?secreting beta?cell line(INS?1 cells) through the orexin receptor 1(OX1R)and the AKT/PKB signaling pathway. Methods INS?1 cells were exposed to different concentrations of orexin A in vitro,and treated with OX1R antagonist(SB334867),PI3K antagonist(wortmannin),or AKT antagonist(PF?04691502). The INS?1 cell proliferation and apoptosis,insulin secretion,OX1R protein activity and AKT phosphorylation level were determined. Results Orexin A(10-10 to 10-6 mol/L)stimulated the proliferation and activation of INS?1 cells,prevented apoptpsis,and increased insulin secretion. Additionally,AKT phosphorylation was stimulated by orexin A(10-10 to 10-6 mol/L). The OX1R antagonist SB334867(10-6 mol/L),the PI3K antagonist wortmannin (10-8 mol/L)and the AKT antagonist PF?04691502(10-6 mol/L)weakened the effects of orexin A. Conclusion Orexin A activated the AKT sig?naling pathway through the mediation of orexin A?OX1R,and promoted cell proliferation in INS?1 cells.

Objective: To explore the effects of 17 β-estrogen (E2) and 2-methoxyestradiol (2ME) on hypoxic induced factor-1α (HIF-1α) and alkane hydroxylase (AlkB) in experimental rats with ovariectomy and hypoxic pulmonary hypertension. Methods: A total of 60 healthy female SD rats with castrated surgery were randomly divided into 6 groups:①Routine oxygen group,②Routine oxygen + E2 group, the rats received subcutaneous injection of E2 (20 μg/kg?d),③Routine oxygen + 2ME group, the rats received 2ME (240 μg/kg?d) and④Hypoxia group,⑤Hypoxia + E2 group,⑥Hypoxia + 2ME group.n=10 in each group and all animals were treated for 8 weeks to establish the hypoxic pulmonary hypertension model. The mean pulmonary artery pressure (mPAP) was measured after bloodletting, right ventricle hypertrophy index (RVHI) was calculated and small pulmonary artery remodeling was observed by HE staining. The expression level of HIF-1α and AlkB were examined by RT-PCR and Western blot analysis. Results: Compared with Routine oxygen group, the rats in Hypoxia group had obviously thickened small pulmonary artery wall with narrowed lumen, increased mPAP and RVHI; the above changes in Hypoxia + E2 and Hypoxia + 2ME groups were relatively smaller, their mPAP and RVHI were higher than Routine oxygen group, while mPAP and RVHI were similar between Hypoxia + E2 and Hypoxia + 2ME groups. There were no real morphological changes in small pulmonary vessels in Routine oxygen + E2 and Routine oxygen + 2ME groups. The HIF-1α expression was obviously elevated in Hypoxia group than Routine oxygen group, while the elevation was less in Hypoxia + E2 and Hypoxia + 2ME groups. HIF-1α expression had no real changes in Routine oxygen+E2 and Routine oxygen + 2ME groups. The AlkB expression was obviously reduced in Hypoxia group than Routine oxygen group, while the reduction was less in Hypoxia + E2 and Hypoxia + 2ME groups. AlkB expression had no real changes in Routine oxygen + E2 and Routine oxygen + 2ME groups. Conclusion: Estradiol E2 and 2ME could remit pulmonary hypertension which might be via up-regulating AlkB expression and down-regulating HIF-1α expression in experimental rats with hypoxic pulmonary hypertension.

BACKGROUND:Although the preparation of bone tissue engineering scaffolds can achieve satisfactory results by solvent casting/particulate leaching, in situ molding method, electrospinning, phase seperation/freeze drying, gas foaming, there are stil some deficiencies in the accuracy, pore uniformity, spatial structure complexity, personalized stents. ＜br> OBJECTIVE:To prepareβ-tricalcium phosphate bone tissue engineering scaffolds using 3D printing. ＜br> METHODS:Drug-loadedβ-tricalcium phosphate scaffolds were prepared with 3D printing, and the structure was observed to measure its porosity and mechanical strength. The scaffold was immersed in simulated body fluid for 15 weeks to observe the quality change. The scaffold was co-cultured with rat bone marrow mesenchymal stem cells for 7 days to observe celladhesion and morphological changes. Rat bone marrow mesenchymal stem cells were cultured in extracts of drug-loadedβ-tricalcium phosphate scaffold and low-glucose Dulbecco's modified Eagle’s medium containing 15%fetal bovine serum for 24, 48, and 72 hours, to determine the absorbance values and cytotoxicity grading, respectively. Meanwhile, the cells were subjected to osteogenic culture for 1 week, and ＜br> the alkaline phosphatase activities in two groups were detected. ＜br> RESULTS AND CONCLUSION:The prepared scaffold showed irregular micropores, high porosity, uniform pore distribution, high pore connectivity rate, and large compressive strength. The drug-loadedβ-tricalcium phosphate scaffold degraded completely with 15 weeks, and cancellous bone defect repair was completed in the same period. Rat bone marrow mesenchymal stem cells adhered to the surface of drug-loadedβ-tricalcium phosphate scaffold and went deep into the scaffold, showing good growth and proliferation. The activity of alkaline phosphatase was also improved. These findings indicate that the drug-loadedβ-tricalcium phosphate scaffold has good biocompatibility.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cisplatin ; administration & dosage ; Cyclophosphamide ; administration & dosage ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fluorouracil ; administration & dosage ; Humans ; Injections, Intra-Arterial ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Phytotherapy

Adult ; Bone Neoplasms ; Humans ; Male ; Nose Neoplasms ; Osteoblastoma ; Skull Base Neoplasms

Retrieval of ancient DNA (aDNA) sequences from organism remains provide direct view of their evolutionary history. However, researches on aDNA have suffered from lots of technical problems. Specifically, discredited sequences were generated from damaged aDNA templates, and expensive and time-consuming methods were employed. Here, a method which could recover the endogenous aDNA as well as to reduce the cost and research period is described. This is achieved by improving the ancient DNA extraction method of isopropanol precipitation, and reevaluating the method of PCR after N-glycosylase (UNG) treatment, which could remove the damaged DNA from the aDNA extract. The efficiency of these methods were tested by comparing with traditional methods using ancient specimens of pig teeth aged between 4 300 years before present (BP) and 3 900 BP. The results showed that: firstly, the extraction efficiency of the improved method of isopropanol precipitation and current method with silica-based spin column were all 60%. Furthermore, the research period at least could be reduced by half with the application of the improved methods and the cost to 1/10 of the current method. Secondly, sequences obtained through the method of PCR after UNG treatment were 100% authentic. In contrast, 66%～ 88% sequences were authentic based on the results obtained with the method of multiple PCRs without UNG treatment. And the research cost and period needed by the method with UNG treatment were only half of the later one. These results demonstrate that the improved extraction method of isopropanol precipitation combined with the method of PCR after UNG treatment could increase the success rate of authentic DNA amplified and at least reduce the research cost and period by half. Therefore, this method can be applied in the large-scale detection of ancient specimens.

BACKGROUND: It remains unclear whether proinflammatory factor, such as tumor necrosis factor-α (TNF-α), γ-interferon and anti-inflammatory cytokine, such as interleukin-10 (IL-10), interleukin-4 (IL-4) levels can change after laryngeal transplantation. OBJECTIVE: To explore the expression of TNF-α and IL-10 in different expressive tissue layers and its relationship during the acute rejection episodes, and to evaluate the role of TNF-α and IL-10 levels in serum for prediction of early acute rejection after laryngeal transplantation. METHODS: Laryngeal heterotopic transplantations were performed in Wistar and Sprague-Dawley rats. According to different dosages of immunodepressant, all recipients were divided into three groups: Group 0 mg, Group 5 mg, and Group 10 mg. Untreated Sprague-Dawley rats served as normal control group. RESULTS AND CONCLUSION: Changes in serum TNF-α and IL-10 levels at postoperation days 3, 7 and 11 were positively correlated with these expressions in the epithelium mucosa and submucosa at various time points after transplantation. These indicate that the high-antigenicity of graft mainly concentrates on the layers of mucosa and submucosa. TNF-α and IL-10 concentrations can serve as indexes for predicting acute rejection after laryngeal transplantation.

ObjectiveTo assess the value of the area of ground glass opacities (GGOs) in lungs displayed by high-resolution computed tomography (HRCT) in paraquat (PQ) poisoned patients in evaluating prognosis. Methods Clinical and imaging data of 137 patients with acute PQ poisoning admitted to Affiliated Hospital of the Medical College of Chinese People's Armed Police Forces from January 2012 to August 2014 were analyzed retrospectively. The plasma concentration of PQ on admission and the area of GGOs were compared between two groups. The lung HRCT within 10 days of poisoning was performed every 3 days, and the areas of GGOs were evaluated on five levels, including aortic arch, aortic pulmonary window, left upper lobe bronchial, right inferior pulmonary vein, and left diaphragmatic dome. Receiver operating characteristic curve (ROC) was plotted to evaluate the value of all the parameters for prognosis.Results Among 137 patients, 45 died within 28 days after poisoning, with the mortality rate of 32.85%. The plasma PQ level in the non-survivors was significantly higher than that in the survivors (mg/L:7.06±0.67 vs. 3.51±0.34,t = 5.280,P = 0.000). The areas of GGOs at three time points in the non-survivors were significantly higher than those in the survivors [1-3 days: (32.0±5.0)% vs. (2.5±0.4)%,t = 7.860,P = 0.000;4-6 days: (45.5±5.7)% vs. (2.8±0.5)%,t = 12.420,P = 0.000; 7-10 days: (68.0±4.8)% vs. (3.0±0.6)%, t = 23.950,P = 0.000]. ROC analysis demonstrated that the area under the ROC curve (AUC) of GGOs in 7-10 days was 1.000, which could be used to determine the prognosis, but it was too late for the treatment. The AUC of GGOs in 4-6 days was 0.979, with the threshold of> 12.0%, the specificity of 96.15%, the sensitivity of 85.19%, the positive predictive value of 88.46%, and the negative predictive value of 94.94%, which presented good effect in predicting prognosis in the early stage of acute PQ intoxication. But plasma PQ concentration was relatively poor for determining prognosis, AUC was 0.821, with the threshold of> 1.95 mg/L, the specificity of 34.52%, the sensitivity of 88.64%, the positive predictive value of 41.49%, and the negative predictive value of 85.29%.Conclusions The area of GGOs displayed by HRCT can be used to evaluate the fully developed acute PQ lung injury, and it is superior to plasma PQ concentration. The area of GGOs displayed by HRCT 4-6 days after intoxication can be used for the evaluation of PQ induced pulmonary injury in the early stage and the evaluation of clinical prognosis.

OBJECTIVE To investigate the spectrum of organisms causing purulent endophthalmitis and to identify the variance of bacterial spectra and their sensitivities to commonly used antibiotics.METHODS The medical records of patients with purulent endophthalmitis receiving pathogen examination in Zhongshan Ophthalmic Center from Jan 2000 to Dec 2006 were analyzed retrospectively.RESULTS Of the 416 cases,82.0% purulent endophthalmistis were caused by ocular trauma and the organisms were isolated in 181(43.5%)of them.Among the pathogens isolated,14.9%,47.5%,9.9% and 27.6% were fungi,Gram-positive cocci,Gram-positive bacilli and Gram-negative bacilli,respectively.The fungal endophthalmistis was sporadic per annum.The most common bacterial pathogen was staphylococcus epidermidis(33.1%) which increased year by year.The most common Gram-negative bacterium was Escherichia coli(10.5%).Among the common pathogens,the Gram-positive cocci were sensitive to fluoroquinolones and cefoperazone,and the Gram-positive bacilli were sensitive to aminoglycosides and cefoperazone while the Gram-negative bacilli were resistant to various kinds of antibiotics in different degree.CONCLUSIONS The purulent endophthalmistis should be monitored in patients with open ocular injury.Fluoroquinolones with cefoperazone to treat the ocular inflammation are a preferred therapeutic regimen for patients with bacterial endophthalmitis with unknown etiopathogenesis.

Background and purpose: Urokinase-type plasminogen activator receptor is related to invasion and metastasis of tumor. Inhibition of uPAR expression in tumor cells results in reducing its metastasis. This study was aimed to construct an expression vector with short hairpin RNA (shRNA) of uPAR, which could pave the way for RNAi-mediated tongue squamous cell carcinoma therapy. Methods: Genome sequences of uPAR gene were retrieved from Genhank and cDNA was designed to code expression of shRNA for uPAR gene. The cDNA was synthesized and inserted into the eukaryotic expression vector pWH1, and the recombinant pWH1-uPAR expression vector was identified by enzyme cutting method. Then, pWH1-uPAR expression vector was transfected into tongue squamous cell carcinoma Ts cells by Lipofectomine 2000. At last, the expression of uPAR in Ts cells transfected with pWH1-uPAR expression vector was observed by RT-PCR, immunocytochemistry staining and Western blot. MTT assay was performed to measure the proliferation of Ts cell. Results: The uPAR shRNA eukaryotic expression vector was successfully constructed. Compared with Ts cells and Ts cells transfected with plasmid pWH1, the Ts cells transfected with pWHI-uPAR expression vector showed a lower mRNA and protein expression of uPAR. The inhibition rate of proliferation was 32.9% of Ts cells by transfected with pWHl- uPAR. Conclusion: The constructed uPAR shR.NA expression vector could inhibit the expression of uPAR in tongue squamous cell carcinoma, which may be helpful for further research on the function of uPAR and provide effective methods for therapy of tongue squamous cell carcinoma.