Repair effects of bone marrow mesenchymal stem cells on benzene-induced hematopoietic injury in mice / 中国职业医学

OBJECTIVE: To explore the repair effects of bone marrow mesenchymal stem cells( BMSCs) on hematopoietic injury induced by benzene poisoning in mice. METHODS: Five specific pathogen free healthy male Kunming mice were selected to obtain BMSCs through bone marrow attachment culturing method. The Kunming mice were randomly divided into poisoning group and BMSCs transplantation group,18 mice in each group,after the benzene poisoning model was established by subcutaneous multi-point injection of benzene and oil mixture 3 times/week,10 weeks continuously. Each group was injected through tail vein with 250. 0 μL 0. 9% sodium chloride solution or 250. 0 μL BMSCs suspension( cell density 2 × 109/L) once per week for 4 weeks,respectively. The control group( 10 mice) was not given any treatment.Mice were euthanized 2 weeks after treatment. The blood routine examination was conducted. Nucleated cells in bone marrow were observed after Giemsa staining. The clones of hemopoietic progenitor cells were counted and the levels of serum interferon-γ( IFN-γ) were examined using enzyme-linked immune sorbent assay. RESULTS: The mouse model of chronic benzene poisoning was established successfully. After the BMSCs transplantation treatment,the white blood cell count,platelet count,red blood cell count,hemoglobin level and bone marrow nucleated cell as well as granulocyte-macrophage colony forming unit( CFU-GM) in benzene poisoning group were significantly decreased compared with control group( P <0. 01),while those indexes of BMSCs treatment group were higher than that of benzene poisoning group( P < 0. 05). The counts of platelet,red blood cell,bone marrow nucleated cell and CFU-GM in BMSCs treatment group were significantly lower than that of control group( P < 0. 05). The level of serum IFN-γ in benzene poisoning group was higher than that of control group( P < 0. 01),and serum IFN-γ level in BMSCs treatment group was lower than that of benzene poisoning group( P < 0. 01). There was no significant difference of IFN-γ level in BMSCs treatment group compared with control group( P > 0. 05). CONCLUSION: BMSCs have repair effects on hematopoietic system injury caused by benzene poisoning.

Application of transthoracic echocardiography in diagnosis of infective endocarditis / 中国医学影像技术

Objective To assess the clinic value of transthoracic echocardiography in diagnosis of infective endocarditis.Methods Transthoracic echocardiography features and clinic data of 114 patients with infective endocarditis were analyzed retrospectively.The form,activity and coaptation of cardiac valve were observed.The vegetation was found,and the surrounding structure of valve and fundamental heart disease were studied.Results Among 114 patients,the primary symptom presented as fever in 104 patiens.Fundamental heart disease was found in 69 patients;and systemic disease was found in 5 patients.Blood culture was positive in 35 patients.Mitral valve was involved in 46 patients.Aortic valve was involved in 48 patients.Tricuspid valve was involved in 17 patients.Pulmonary valve was involved in 1 patient.Prosthetic valve was infected in 9 patients.Multi-valves were infected in 7 patients.There were serious complications in 19 patients.Ultrasound showed vegetation in 56 of 59 patients underwent surgery.And the diagnostic accuracy rate was 94.92％ (56/59).Conclusion Transthoracic echocardiography can be used to accurately diagnose infective endocarditis.

Cloning, expression and characterization of gene encoding human stem cell growth factor-alpha and its synergetic effect with rhGM-CSF on proliferation of human umbilical cord mesenchymal stem cells / 生物工程学报

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.

Isolation of human umbilical cord mesenchymal stem cells and differentiation into adipocytes and osteblasts / 中国组织工程研究

BACKGROUND:Culture condition,isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells,which lack of unified identification standards.Therefore,it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem calls(hUCMSCs).OBJECTIVE:To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts.METHODS:The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase.The morphology,proliferation and immunophenotype of the 3rd passage cells were analyzed,and then cells were induced to osteogenic and adipogenic differentiation in vitro.RESULTS AND CONCLUSION:The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro.Flow cytometry analysis revealed that the hUCMSCs were positive for CD29 CD44,CD59,CD105,but were negative for CD40,CD86 and HLA-DR.These calls could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions.The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro.This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts.

Expression, purification and activity determination of cyanovirin-N / 生物工程学报

Cyanovirin-N (CVN) is an 11 kDa anti-HIV protein originally isolated from extracts of a cyanobacterium, Nostoc ellipsosporum. The protein binds with high affinity to the viral envelope glycoprotein gp120 and irreversibly inactivates diverse HIV strains. A fusion gene consisting of cvn, sumo and 6xHis tag was synthesized by PCR according to the codon bias of Escherichia coli. The fusion protein is expressed in the cytoplasm of E. coli in a soluble form and up to 28% of the total protein. The recombinant CVN was purified to homogeneity by 2 rounds of Ni-NTA affinity chromatography and one round of SUMO protease cleavage. Bioactivity assay demonstrated that SUMO-CVN and CVN bound to gp120 with nanomolar concentration. In addition, CVN showed potent anti-HSV-1 and anti-HIV-1 activities in in vitro cellular assays. Therefore, the 6xHis SUMO fusion expression and purification system provides a better approach for large scale production of CVN for further microbicide development.

Incidences of malnutrition and abdominal obesity in elderly type 2 diabetes / 中华临床营养杂志

Objective To investigate the incidences of malnutrition (including undernutrition, overweight, and obesity) and abdominal obesity in elderly type 2 diabetes. Methods Totally, 133 elderly type 2 diabetes patients [study group, aged (66.9 5±.4) years] and 133 age-matched healthy subjects [control group, aged (66. 3 ±5.8) years] who met entry criteria and obtained informed consent were randomly enrolled into this study. Body weight, total body fat (TBF), abdominal fat, visceral fat, visceral fat area, and waist-to-hip ratio (WHR) were measured by multi-frequency bioelectric impedance analysis. The incidences of undernutrition, overweight, obesity, and abdominal obesity judged by BMI and WHR respectively were compared between the two groups. Results Com- pared to control group, BMI [(25.7 3.8) vs. (24.2 2.2) kg/m2, P = 0.001 ], TBF [ (20.1±6.9) vs. (17.4 5.0) kg, P = 0.001], WHR (0. 92±0.10 vs. 0.87±0.06, P =0.001), abdominal fat [(10.2 3.4) vs. (8.6 2.5)kg, P= 0.001], visceral fat [(2.7±0.9) w. (2.3 0.7)kg, P =0.001 ], and visceral fat area [ (89.1±28. 8) vs. (75. 7±21. 6) cm2, P =0.001 ] significantly increased in study group. The incidences of undernutrition (BMI<18.5, 3.8% vs. 0, P=0.024) and obesity (BMI

Inhibitory effects of nm23-H1 gene on proliferation and invasion of A549 cell line / 中国病理生理杂志

AIM: To study the inhibitory effects of nm23-H1 gene on proliferation and invasion of human lung adenocarcinoma A549 cell line. METHODS: Recombinant eukaryotic expression vector pcDNA3.1-nm23-H1 containing full length of human nm23-H1 cDNA was constructed and transfected into a human lung adenocarcinoma A549 cell line by lipofectamine. Cell strain that expressed nm23-H1 stably was screened out by G418 and named pcDNA-nm23-A549. Expression of nm23-H1 was identified by RT-PCR and immunohistochemistry. Growth curves were drawn to detect the inhibitory effects on cell proliferation. Cell cycle of pcDNA-nm23-A549 was examined by flow cytometry. Atomic force microscopy was used to observe the filopodia on the surface of the cells. RESULTS: Introduction of nm23-H1 obviously inhibited the proliferation of A549. Expression of nm23-H1 did not induce apotosis in A549 cells but increased the percentage of phase G_1 cells and decreased phase S cells. Meanwhile, phase G_1 to phase S transition was restrained. Filopodia in the cell surface was much fewer and its structure changed in cells transformed. CONCLUSION: nm23-H1 is capable of inhibiting A549 proliferation and decreasing its metastatic ability, probably by interfering with cell cycle and cell surface structure.

Recent advances in angiogenesis and tumor angiogenesis inhibitor in prostate cancer / 中国病理生理杂志

A Review Angiogenesis plays a key role in progression of prostate cancer. Antigiogenesis becomes a new treament target for prastate cancer. In this review, we focus on the current knowledge of angiogenesis and tumor angiogenesis inhibitor in prastate cancer. [

A Case Study of career Identity Development / 中国临床心理学杂志

Objective: Exploring the developmental vitial moments and process of career identity.Methods: A case subject was administered structured interview and tested by Career Identity and the Related Factors Questionnaire,Chinese Self-Efficacy Scale,QZPS and MBTI-G.Results: There were several vital moments in the process of career identity,such as the first time to think about career identity,and select a major,the first time to choose career and get employed,meet the organizational changes,find a new job again and get promotion.The present crisis of career identity reflected some unresolved problems of previous developmental periods.Conclusion: The individual's objective behavior toward career choice,decision-making,continuous exploration and efforts were of great benefit to career identity.

Expression of recombinant human tumor suppressor NDPK-A in E. coli / 中国病理生理杂志

AIM: To construct E. coli expression plasmid of recombinant human NDPK-A with a 6?His tag, optimize the expression condition and identify the activity of the product. METHODS: nm23-H1 was subcloned from plasmid pBVNMH1 to pQE40 which contain 6?His purification tag. The expression condition was modulated in grades to get the optimal expression. We purified protein with the Ni+-NTA affinity chromatography column, identified the immunogenicity of the product with Western blot, and measured the kinases activity with HPLC. In addition, angiogenesis inhibition activity of rhNDPK was identified by CAM. RESULTS: The sequence of nm23-H1 subclone in pQE40 was exactly correct. The expression rate of rhNDPK-A was 49 6%. Purified rhNDPK-A specially recognized the antiserum of NDPK-A. It also inhibited angiogenesis. CONCLUSION: PQE-nm23H1 containing 6?His can express target protein at high level. This purification method is simple than other methods, and the product has the same activity as natural human NDPK-A.