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.

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.

Objective To explore the efficacy of hormone replacement therapy (HRT) for rheumatoid arthritis (RA).Methods The literature about HRT in the treatment of rheumatoid arthritis were searched.Eleven papers were subjected to a Meta-analysis and a heterogeneity test was conducted to evaluate the efficacy of HRT.Results HRT reduced the level of erythrocyte sedimentation rate (ESR) in RA patients (P=0.016),the SMD was-0.22(-0.40,-0.04); improved bone mineral density (BMD) in RA patients (P=0.022),the WMD was 2.83(0.41,5.26); decreased clinical parameters for disease activity evaluations of RA patients (P=0.048),the SMD was-0.19 (-0.38,0.00); decreased the level of C-reactive protein (CRP) in RA patients,the SMD was-0.08 (-0.37,0.21),but the difference was not statistically significant (P=0.591).Conclusion The findings from this Meta-analysis indicate that HRT can reduce the ESR level,improve clinical indexes and improve BMD level of RA patients.HRT may suppress disease activity and osteoporosis of RA patients,so it may be used as an auxiliary therapy in the treatment of RA.

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.

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.

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.

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

Objective To investigate the efficacy and safety of amisulpride and clozapine in schizophrenia patients with predominantly negative symptoms.Methods Totally 166 cases of schizophrenia patients with predominantly negative symptoms from May 2013 to May 2016 in The Sixth People's Hospital of Hebei were divided into observation group and control group,83 cases in each group.Patients in the observation group were treated with amisulpride,and control group were treated with clozapine.The clinical effect,SANS scores,and occurring rate of abnormal electrocardiogram were compared.Results The clinical effect,emotional insipid (blunting),and attention dysfunction scores from SANS of observation group were significantly better than those of control group on week 4,8,and 12,respectively (P ＜ 0.05);The occurring rate of abnormal electrocardiogram in observation group was significantly lower than that of control group on week 12 (P ＜ 0.05).Conclusion Compared with clozapine,amisulprid has better efficacy and safety in schizophrenia patients with predominantly negative symptoms,and can effectively improve of symptom of insipid (blunting) and attention dysfunction.