Effect of Wnt/β-catenin signaling pathway in neural differentiation of human bone marrow mesenchymal stem cells / 中国修复重建外科杂志

OBJECTIVE@#To explore the effect of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and the combination of bFGF and EGF in the neural differentiation of human bone marrow mesenchymal stem cells (hBMSCs), and the role of Wnt/β-catenin signaling pathway in this process.@*METHODS@#The identified 4th-generation hBMSCs were divided into five groups according to different induction conditions, namely control group (group A), EGF induction group (group B), bFGF induction group (group C), EGF and bFGF combined induction group (group D), and EGF, bFGF, and Dickkopf-related protein 1 (DKK-1) combined induction group (group E). After 7 days of continuous induction, the cell morphology was observed by inverted fluorescence phase contrast microscopy, levels of genes that were related to neural cells [Nestin, neuron-specific enolase (NSE), microtubule-associated protein 2 (MAP-2), and glial fibrillary acidic protein (GFAP)] and key components of the Wnt/β-catenin signaling pathway (β-catenin and Cyclin D1) were detected by RT-PCR, and the levels of proteins that were related to neural cells (Nestin and GFAP) as well as genes that were involved in Wnt/β-catenin signaling pathway [β-catenin, phosphorylation β-catenin (P-β-catenin), Cytoplasmic β-catenin, and Nuclear β-catenin] were explored by cellular immunofluorescence staining and Western blot.@*RESULTS@#When compared to groups A and B, the typical neuro-like cell changes were observed in groups C-E, and most obviously in group D. RT-PCR showed that the relative expressions of Nestin, NSE, and MAP-2 genes in groups C-E, the relative expressions of GFAP gene in groups D and E, the relative expression of NSE gene in group B, the relative expressions of β-catenin gene in groups C and D, and the relative expressions of Cyclin D1 gene in groups B-D significantly increased when compared with group A ( P<0.05). Compared with group E, the relative expressions of Nestin, NSE, MAP-2, GFAP, β-catenin, and CyclinD1 genes significantly increased in group D ( P<0.05); compared with group C, the relative expression of Nestin gene in group D significantly decreased ( P<0.05), while NSE, MAP-2, and GFAP genes significantly increased ( P<0.05). The cellular immunofluorescence staining showed that the ratio of NSE- and GFAP-positive cells significantly increased in groups C-E than in group A, in group D than in groups C and E ( P<0.05). Western blot assay showed that the relative expression of NSE protein was significantly higher in groups C and D than in group A and in group D than in groups C and E ( P<0.05). In addition, the relative expression of GFAP protein was significantly higher in groups C-E than in group A and in group D than in group E ( P<0.05). Besides, the relative expressions of β-catenin, Cytoplasmic β-catenin, Nuclear β-catenin, and the ratio of Nuclear β-catenin to Cytoplasmic β-catenin were significantly higher in groups C and D than in group A and in group D than in group E ( P<0.05), whereas the relative expression of P-β-catenin protein was significantly lower in groups C and D than in group A and in group D than in group E ( P<0.05).@*CONCLUSION@#Different from EGF, bFGF can induce neural differentiation of hBMSCs. In addition, EGF can enhance the hBMSCs neural differentiation of bFGF, while the Wnt/β-catenin signaling pathway may play a positive regulatory role in these processes.

Study on NAEK-labelled HiD-Hin47 fusion protein as a carrier for pneumococcal polysaccharide conjugate vaccine / 中华微生物学和免疫学杂志

Objective:To develop an unnatural amino acid-labelled HiD-Hin47 fusion protein as a novel carrier.Methods:Twenty versions of the fusion protein were designed, each of which contained a different amino acid site replaced by an azide-bearing amino acid, N6-(2-azidoethoxy) (carbonyl)-L-Lysine (NAEK). These fusion proteins were constructed, expressed and purified, and the yield were evaluated by SDS-PAGE. Based on the highest protein yield, which was approximately 70% of the wild-type yield, the fusion protein with the unnatural amino acid in E677 site was selected. The pneumococcal polysaccharides of serotype 3 (F3) and serotype 6B (F6B) were coupled to the selected fusion protein through a "click" reaction. The conjugates were purified and compared in animal studies with other F3 and F6B conjugates that were coupled to CRM197, tetanus toxoid (TT) and HiD by conventional methods.Results:The immunogenicity of F3 conjugate using HiD-Hin47 as carrier (F3-HiD-Hin47) was slightly better than that of other F3 conjugates. F6B-HiD-Hin47 was significantly better than F6B-TT and F6B-HiD in terms of immunogenicity, but showed no significant difference with F6B-CRM197.Conclusions:NAEK-labelled HiD-Hin47 had the potential as a carrier for pneumococcal polysaccharide conjugate vaccine and was worthy of further study.

Application of upper arm totally implantable venous access ports in 34 patients with tumor / 中华临床营养杂志

Objective To investigate the safety,feasibility and clinical application effect of upper arm totally implantable venous access port (TIVAP) inserted by the combination of oncologists and nurses in patients with tumor.Methods A total of 34 patients,who needed long-term transfusion treatment,were included in this study with upper arm TIVAP from March 2017 and December 2017.There were 20 males and 14 females.The median age was 63 (35 ～ 83) years.Upper arm TIVAP was implanted by both doctors and nurses into the patients with tumor,and the TIVAP related success rate,complications and patients satisfaction were recorded.Results 34 patients all succeeded in TIVAP implantation with the operation success rate of 100％.The average operation time was about 40 minutes (30 to 60 minutes) from the disinfecting cloth to the end of the suture.The operations of all patients were successful.After the operation two patients died of cancer progression,one patient had soft tissue infection around capsular bag,None of the patients had other complications such as blocked infusion,catheter shift,port reversal,and so on,and the incidence of complication was 2.94％ (1/34).Conclusions Upper arm TIVAP has the advantages of safe puncture,shorter operation time,few intra-operative and post-operative complications and higher feasibility for operation by both oncologist and nurse,which can supplement the limitations and deficiencies of the chest wall port and PICC in a certain extent,therefore is a good choice for clinical application.

Characteristics and risk factors analysis of cognitive decline in elderly patients with cerebral infarction / 中国脑血管病杂志

Objective To investigate the characteristics and risk factors for cognitive decline in elderly patients with cerebral infarction.Methods A cross-sectional study was performed to investigate the population of Jiang'an Town,Rugao,Jiangsu Province from November 13,2014 to December 21,2014.The samples from the Rugao longitudinal study of aging included 1 788 individuals from 31 villages aged from 70 to 84 years.They were all Han nationality,including 830 males and 958 females.History of cerebral infarction was identified according to the neurological diagnosis confirmed by the secondary hospital and above or brain CT.The modified Kyohko Hasegawa dementia scale was used to evaluate the cognitive function.Five dimensions (orientation,memory,near memory,computing power,and common sense) of this population were assessed.The total score >21.5 was non-cognitive impairment and ≤21.5 was cognitive impairment.Multivariate logistic regression was used to analyze the risk factors for cognitive decline in elderly patients with cerebral infarction.Results (1) In the 1 788 subjects,133 had cerebral infarction (7.4%),and 1 655 did not have cerebral infarction.The proportion of hypertension in patients with cerebral infarction was higher than that without cerebral infarction (63.9% [n=85] vs.41.7% [n=690]).The high-density lipoprotein cholesterol level was lower than that of the non-cerebral infarction group (1.40±0.29 mmol/L vs.1.47±0.33 mmol/L).There was statistically significant difference (all P<0.05).(2) The patients with cerebral infarction were partially impaired in orientation and computational power,and the overall cognitive function score was 20±7,which was significantly lower than patients with non-cerebral infarction (21±6).The difference between the two groups was statistically significant (P<0.05).(3) In 133 patients with cerebral infarction,76 had cognitive impairment,the incidence was 57.1%,and 59 of them were women.The average value of serum creatinine in patients with cognitive impairment was 59±15 μmol/L,which was significantly lower than those with non-cognitive impairment (66±14 μmol/L).There was significant difference (P<0.05).(4) Multivariate logistic regression analysis showed that the education level below primary school (OR,2.86,95%CI 2.19-3.72) and female (OR,1.85,95%CI 1.50-2.28) were the independent risk factors for cognitive decline in elderly patients with cerebral infarction.High serum creatinine concentration (OR,0.96,95%CI 0.95-0.97) was a protective factor for it.Conclusion The cognitive function of the elderly was decreased after cerebral infarction,especially in the aspect of orientation and calculation.The education level below primary school and women were the independent risk factors for cognitive impairment,and high serum creatinine concentration had a certain protective effect.

Association between ACTN3 gene R577X polymorphism and blood lipid levels in an elderly Chinese Han population in Rugao,China / 复旦学报（医学版）

Objective To explore the association between ACTN3 gene R577X polymorphism and serum lipid levels in an elderly Chinese Han population.Methods This study was based on the ageing arm of The Rugao Longevity and Ageing Study (RtLAS).Genotyping was performed by Taqman MGB method.Lipids includedtotal cholesterol (TC),triglyceride (TG),high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C).Cut points of dyslipidemia were based on that reported in the NCEP ATPIII.We explored the associations between R577X polymorphism loci and dyslipidemia by using analysis of covariance and logistic regression analysis.Results Overall 1 618 samples was included (741 males and 877 females) in this study.Covariance analysis found that TC level of RR,XX and RX genotype in the general population were 1.34,1.37,1.43 mmol/L with an increasing trend (P =0.024);TC and LDL-C levels of RR,XX and RX genotype (TC:5.13,5.29,5.43 mmol/L,P =0.004;LDL-C:2.76,2.88,3.00 mmol/L,P =0.004) were significantly different in females.Logistic regression analysis showed that the increased copies of X allele were significantly associated with increased TC and LDL-C levels in the general population and females.For the general population,TC:OR=1.184,95％CI:1.030-1.361,P =0.018;LDL-C:OR =1.334,95％CI:1.101-1.588,P=0.003;For females,TC:OR=1.332,95％ CI:1.102-1.616,P =0.003;LDL-C:OR =1.549,95 ％CI:1.208-1.986,P =0.001.After adjusting for other covariates,the above associations remained significant.Conclusions ACTN3 gcnc R577X polymorphism is associated with plasma TC and LDL-C levels in elderly woman of Han populations in Rugao,China.

Analysis of influencing factors of undergraduate teaching quality and countermeasures / 中华医学教育探索杂志

An overall improvement of teaching quality is the core task of the reform and the development of higher education. Based on the analysis of the main factors of affecting the teaching quality in universities and combined with the implementation of “views on improving the overall qual-ity of higher education”by the Education Ministry, countermeasures for improving the teaching quality of undergraduates have been proposed such as increasing higher education funds investment to im-prove the condition of running a school, paying attention to the construction of teaching management staff to improve the level of teaching management, establishing Teacher Development Research Center to consolidate the fundamental position of undergraduate teaching , utilizing the high-quality social resources to implement the collaborative education innovation, and improving the personnel training system to strengthen the teaching quality standard .

Proliferation, senescence and differentiation of mesenchymal stem cells:canonical and non-canonical regulations of Wnt signaling pathway / 中国组织工程研究

BACKGROUND:As mesenchymal stem cells are commonly used as seed cells in studies of regenerative medicine and tissue engineering, the regulatory mechanism of their biological characteristics is a current research focus. OBJECTIVE:To summarize the regulations of Wnt signaling pathway on proliferation, senescence and differentiation of mesenchymal stem cells. METHODS:PubMed database and CNKI database were retrieved by computer using the key words of“mesenchymal stem cells, Wnt signaling pathway, proliferation, senescence, differentiation”in Chinese and English, respectively, between 2002 and 2014. Final y, 44 articles were included in result analysis. RESULTS AND CONCLUSION:Wnt signaling pathway is widely involved in the regulations of the biological characteristics of mesenchymal stem cells. Canonical Wnt signaling pathway reveals a bi-directional regulation effect on cellproliferation and osteogenic differentiation, and enhances senescence and neural differentiation, but inhibits adipogenic differentiation;non-canonical Wnt signaling pathway enhances senescence and osteogenic differentiation, and inhibits proliferation and adipogenic differentiation of mesenchymal stem cells, but it takes no part in neural differentiation of mesenchymal stem cells. So the regulations of Wnt signaling pathway on the biological characteristics of mesenchymal stem cells can be used as the new therapeutic targets of bone tissue engineering, nerve injury repair, and so on.

Effect of quercetin combined with cisplatin on proliferation and apoptosis of human osteosarcoma cell line MG-63 / 中国药理学通报

Aim To investigate the effect and mecha-nism of quercetin combined with cisplatin on prolifera-tion and apoptosis of human osteosarcoma cell line MG-63 . Methods MG-63 cells were treated with quercetin alone or combined with cisplatin. Cellular morphologic changes were observed under inverted phase contrast microscope. The effects of proliferation inhibition were assayed by CCK-8 method. The combination effect was judged through Chou-Talaly analysis. The apoptosis ra-tios of cells were analyzed by flow cytometry. The gene expression of Bcl-2 and caspase-3 was detected by RT-PCR assay. The protein expression of Bcl-2 and caspase-3 was measured by Western blot assay. Re-sults Quercetin alone or combined with cisplatin could inhibit the proliferation, but induce the apoptosis of MG-63 cells. Combination of quercetin and cisplatin revealed a synergistic effect on cell proliferation and apoptosis as it reduced the expression of Bcl-2 but en-hanced that of caspase-3 at both gene and protein lev-els. Conclusion Synergistic effect of quercetin com-bined with cisplatin on cell proliferation and apoptosis of MG-63 cells is possibly due to reduction of Bcl-2 and enhancement of caspase-3 expression.

Measure Analysis and Practical Thinking of Curriculum Quality Evaluation / 中华医学教育探索杂志

Curriculum quality evaluation is a connecting medium of building internal quality guaranty system and external Teaching evaluation system, It is an important means of propelling connotative construction and accelerating quality upgrade. This paper introduces the construction principle,executive plan and practical evaluation result of curriculum quality evaluation, and conducts information comparison and statistical analysis from three levels of macrocosm, mid-scope and microcosm. The problem of evaluation index and the adjusting strategy of evaluation system are inspected during the practice.