Objective·To establish a cell-based screening system for identification of compounds with activity in regulating retinoic acid receptor (RARα) stability. Methods·The modified pMSCV plasmid constructs, named as RARα-EGFP-IRES-DsRed, consists of enhanced green fluorescent protein (EGFP) fusing to RARα and red fluorescent protein (DsRed) as internal references incorporating the internal ribosome entry site (IRES) as interval sequence. The RARα-EGFP-IRES-DsRed plasmid was stably transfected into NB4 cells which were named as NB4-pMGIR-RARα. Fluorescence signals of EGFP and DsRed indirectly reflecting the expression of RARα, were detected by flow cytometry in cells that were treated with all-trans retinoic acid, sodium valproate, cytarabine, lenalidomide, etoposide, montelukast and gambogic acid, respectively. Effects of these compounds on the expression of RARα protein were further examined by Western blotting. Results·A double fluorescence reporter system for screening compounds that can increase the stability of RARα protein was successfully established, and sodium valproate was identified as a potent compound to promote the stability of RARα. Conclusion·The double fluorescence reporter system can be used to screen compounds regulating the stability of RARα protein, which can be further used to identify compounds regulating the stability of other proteins.

BACKGROUND:The limitations of computer technology in the study of bone biomechanics and the prediction of bone fixation strength, stability, fatigue damage and life expectancy are more difficult. OBJECTIVE:To investigate the new progress and application of finite element analysis in orthopedics. METHODS:The first author searched PubMed (http://www.ncbi.nlm.nih.gov/PubMed) and CNKI China journal ful-text database (http://www.cnki.net/) published til November 2015. Key words were“finite element analysis, orthopedics, biomechanics”. There were 51 references in English and 320 Chinese literatures. According to the inclusion criteria, 40 literatures were selected. RESULTS AND CONCLUSION:Biomechanics of human skeleton is very complex, and most of the mechanical state is a locomotive, non-static process, thus increasing the difficulty of orthopedic biomechanics research. The prediction concerning bone fixation strength, stability, fatigue damage and lifetime is more difficult. However, the finite element analysis technology, which has been widely applied and demonstrated its reliability actual y in engineering fields, can solve these problems effectively. With the rapid development of computer technology, finite element analysis in the field of orthopedic applications has increasingly been used, which also promoted the development of orthopedic technology.

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.

[ ABSTRACT] AIM: To detect basic fibroblast growth factor ( bFGF ) expression in clinical common malignant tumor ( non-small-cell lung cancer,breast cancer, colon cancer and melanoma) , and to identify relationship between the expression and tumor clinicopathological characteristics.METHODS:Immunohistochemical SP method was used to detect the expression of bFGF at protein level in 208 cases of paraffin-embedded tissue of primary malignant tumor patients ( 68 cases of lung cancer, 80 cases of breast carcinoma, 41 cases of colon cancer and 19 cases of melanoma) .RESULTS:The bFGF protein expression levels were significantly higher in low differentiated non-small-cell lung cancer with lymph node metastasis, and were positively correlated with TNM.In addition, no significant influence of the bFGF protein expression on the patients with median survival period was observed.The protein expression of bFGF was higher in advanced breast cancer with lymph node metastasis and was commonly found in the middle/higher differentiated colon cancer with regional lymph node metastasis.Meanwhile, bFGF protein was highly expressed in advanced melanoma patients with lymph node metastasis.CONCLUSION:bFGF may participate in the process of occurrence and progression of malignant tumor.Ex-pression of bFGF protein may be an effective parameter for evaluating metastasis and prognosis of malignant tumor.

Objective To observe the effects of melatonin (MT) on the expression of heme oxygenase-1 (HO-1),phosphorylated adenine dinucleotide quinone oxidoreductase-1 (NQO-1) and nuclear factor erythroid-2-related factor 2 (Nrf2),so as to explore the mechanism of MT's action in the Nrf2-ARE signaling pathway.Methods A total of 72 Sprague-Dawley rats were randomly divided into a control group,an injury group and a melatonin group,each of 24.T11-T12 acute SCI was induced in the injury and melatonin groups using the modified Allen's method.Ten minutes after the injury,equal amounts of absolute ethyl alcohol and melatonin were intraperitoneally injected into the rats in the injury and melatonin groups.For the control group,the vertebral plate was cut to expose the T11-T12 spinal cord without any injury of the nerves.Six rats from each group were randomly selected for sacrifice at 6,12 and 24 hours after the operation,and T11-T12 spinal cord specimens were collected.The spinal cord injury and inflammatory response were observed using haematoxylin eosin staining.The expression of HO-1,NQO-1 and Nrf2 was examined using immunofluorescence,while the expression of HO-1,NQO-1 and Nrf2 protein and mRNA were detected using RT-PCRs.Results The neuronal cells in the spinal cords of the control rats were of normal shape,without edema,necrosis or obvious hemorrhagic foci.Hemorrhagic foci,significantly more inflammatory cells and some spinal cord neurons with edema and necrosis were observed in the injury group.However,significantly fewer hemorrhagic spots and cells with edema were found in the melatonin group compared with the injury group.The average expression of HO-1,NQO-1 and Nrf2 protein and mRNA was significantly higher in the melatonin group than in the other two groups.The levels in the injury group were also significantly higher than in the control group 12 and 24 hours after the experiments.Immunofluorescence showed that the greatest number of cells with HO-1,NQO-1 and Nrf2 was found in the melatonin group,followed by the injury group and then the control group,with significant differences among all 3 groups.Conclusion Melatonin can promote the expression of HO-1,NQO-1 and Nrf2 in rats with acute spinal cord injury,which might be related with its activating the Nrf2-ARE signaling pathway.

Objective:To evaluate the changes of quality of life in patients with axial spondyloarthritis (ax-SpA) after treatment with non-steroidal anti-inflammatory drugs (NSAIDs) by the 36-item Short Form Health Survey (SF-36).Methods: 120 patients diagnosed with ax-SpA were collected in the first Affiliated Hospital of Zhengzhou University from October 2014 to September 2015.They all agreed to be treated with the special drugs and assessed by special scale.Then they all signed the agreement.In the 3 months,double-blind,parallel controlled trial patients were randomized to 200 mg twice daily (bid) imrecoxib,or 200 mg twice daily (bid) celecoxib.They were assessed for the changes of quality of life at enrollment and after three months of NSAIDs therapy by the SF-36 of Chinese edition.The correlation between quality of life and erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),Bath Ankylosing Spondylitis Disease Activity Index (BASDAI),Bath Ankylosing Spondylitis Functional Index (BASFI),Spondylo Arthritis Research Consortium of Canada (SPARCC) was analyzed.Results: A total of 116 ax-SpA patients completed the study and 4 patients were lost to follow-up.We used the SF-36 scale to assess the quality of life in patients with ax-SpA before and after 3 months,NSAIDs treatment.The treatment effects were not statistically significant difference between the two drugs (P>0.05).After all the patients were treated with NSAIDs for 3 months,there was statistically significant difference (P<0.05) of the physical functioning,role-physical,bodily pain,general health,social functioning,role-emotional;and there was no statistically significant difference (P>0.05) of vitality and mental health.The positively significant correlations had been identified between BASDAI and PF,RP,BP,GH,VT,SF,RE (P<0.05),while no significant correlation was found between BASDAI and MH (P>0.05).A positively significant correlation had been identified between BASFI and PF,RP,BP,GH,SF,RE,MH (P<0.05),while no significant correlation was found between BASFI and VT (P>0.05).The ESR was positively correlated with SF,RE (P<0.05);and CRP was positively correlated with SF,MH (P<0.05);and SPARCC was positively correlated with PF (P<0.05).BASDAI and BASFI were the important influence factors of PF (P<0.05);and BASDAI was the important influence factor of BP,GH,VT,RE(P<0.05);BASFI was the important influence factor of RP,SF,MH(P<0.05).Conclusion: Non-steroidal anti-inflammatory drugs can improve the quality of life of the ax-SpA patients.Imrecoxib and celecoxib have the equivalent curative effect.SF36 scale is suitable for the assessment of the quality of life in patients with ax-SpA.

Objective:To retrospectively analyze the test results of novel coronavirus (2019-nCoV) in different samples (throat swab, sputum and feces) collected from recovered COVID-19 patients in order to provide a more reliable basis for discharge and reduce the risk of recurrence after discharge.Methods:Throat swabs and sputum were sampled in pairs from 78 patients before discharge and sampled in pairs twice from 54 cases with an interval of 1-5 d. Real-time fluorescence quantitative RT-PCR was used to detect the virus in the two types of samples. Throat swab, sputum and fecal samples of six patients were tested for 2019-nCoV during follow-up.Results:The detection rate of viral nucleic acid was 46.15% in throat swabs and 50.00% in sputum samples. Test results of the second paired samples showed that the detection rate of viral nucleic acid was 25.93% in throat swabs and 46.30% in sputum samples, and the difference between the two types of samples was statistically significant ( P<0.05). During follow-up, 2019-nCoV nucleic acid could be detected in the fecal samples of the six patients, but not in their throat swab and sputum samples. Their fecal samples remained positive up to 52 d. Conclusions:In the late convalescence, the respiratory symptoms of COVID-19 patients gradually disappeared with the improvement of clinical symptoms. Moreover, the virus might enter the gastrointestinal tract from respiratory tract, and could long-term exist in recovered patients and be excreted in feces. In order to reduce the rate of missed detection and avoid false negative results, it was suggested to test the viral nucleic acid in different types of samples before a COVID-19 patient was discharged.

Objective:To explore the clinical characteristics, diagnosis and treatment of allergic bronchopulmonary aspergillosis(ABPA) with eosinophilic granulomatous with polyvasculitis(EGPA) as a comorbidity.Methods:We collected the clinical data of a patient with EGPA who sought treatment with ABPA as a comorbidity. We summarized the diagnosis and treatment process of the patient, and reviewed the literature. After that, we discussed the relationship between the pathogenesis of ABPA and EGPA and the diagnosis and treatment experience.Results:A 61-year-old male patient suffered from repeated coughing, expectoration, hemoptysis, wheezing. His blood eosinophils count and immunoglobulin (Ig)E level were elevated. He was tested positive for aspergillus fumigatus. His Computer Tomography (CT) showed pulmonary nodules and bronchiectasis. He was diagnosed as ABPA. He also suffered limb numbness, sinusitis, and renal dysfunction and was diagnosed as EGPA. His condition improved after treatment with glucocorticoids, immunosuppressants and antifungal agents. We reviewed the relevant literature and retrieved 10 case reports, of which 5 cases were diagnosed as ABPA first and then EGPA, 3 cases were diagnosed as EGPA first and then ABPA, 2 cases were diagnosed simultaneously. We found that there was a certain correlation between them in the pathogenesis, and the main treatment is glucocorticoids, immunosuppressants and antifungal drugs.Conclusion:ABPA with EGPA as a comorbidity is rarely reported, which reminds us that when diagnosing one of the diseases in clinical work, we should be alert to the coexistence of another disease to avoid misdiagnosis.

Background: Insulin resistance (IR) is the key pathological basis of many metabolic disorders. Lack of asialoglycoprotein receptor 1 (ASGR1) decreased the serum lipid levels and reduced the risk of coronary artery disease. However, whether ASGR1 also participates in the regulatory network of insulin sensitivity and glucose metabolism remains unknown. Methods: The constructed ASGR1 knockout mice and ASGR1-/- HepG2 cell lines were used to establish the animal model of metabolic syndrome and the IR cell model by high-fat diet (HFD) or drug induction, respectively. Then we evaluated the glucose metabolism and insulin signaling in vivo and in vitro. Results: ASGR1 deficiency ameliorated systemic IR in mice fed with HFD, evidenced by improved insulin intolerance, serum insulin, and homeostasis model assessment of IR index, mainly contributed from increased insulin signaling in the liver, but not in muscle or adipose tissues. Meanwhile, the insulin signal transduction was significantly enhanced in ASGR1-/- HepG2 cells. By transcriptome analyses and comparison, those differentially expressed genes between ASGR1 null and wild type were enriched in the insulin signal pathway, particularly in phosphoinositide 3-kinase-AKT signaling. Notably, ASGR1 deficiency significantly reduced hepatic gluconeogenesis and glycogenolysis. Conclusion The ASGR1 deficiency was consequentially linked with improved hepatic insulin sensitivity under metabolic stress, hepatic IR was the core factor of systemic IR, and overcoming hepatic IR significantly relieved the systemic IR. It suggests that ASGR1 is a potential intervention target for improving systemic IR in metabolic disorders.

Background: Insulin resistance (IR) is the key pathological basis of many metabolic disorders. Lack of asialoglycoprotein receptor 1 (ASGR1) decreased the serum lipid levels and reduced the risk of coronary artery disease. However, whether ASGR1 also participates in the regulatory network of insulin sensitivity and glucose metabolism remains unknown. Methods: The constructed ASGR1 knockout mice and ASGR1-/- HepG2 cell lines were used to establish the animal model of metabolic syndrome and the IR cell model by high-fat diet (HFD) or drug induction, respectively. Then we evaluated the glucose metabolism and insulin signaling in vivo and in vitro. Results: ASGR1 deficiency ameliorated systemic IR in mice fed with HFD, evidenced by improved insulin intolerance, serum insulin, and homeostasis model assessment of IR index, mainly contributed from increased insulin signaling in the liver, but not in muscle or adipose tissues. Meanwhile, the insulin signal transduction was significantly enhanced in ASGR1-/- HepG2 cells. By transcriptome analyses and comparison, those differentially expressed genes between ASGR1 null and wild type were enriched in the insulin signal pathway, particularly in phosphoinositide 3-kinase-AKT signaling. Notably, ASGR1 deficiency significantly reduced hepatic gluconeogenesis and glycogenolysis. Conclusion The ASGR1 deficiency was consequentially linked with improved hepatic insulin sensitivity under metabolic stress, hepatic IR was the core factor of systemic IR, and overcoming hepatic IR significantly relieved the systemic IR. It suggests that ASGR1 is a potential intervention target for improving systemic IR in metabolic disorders.