Objective To observe the effects of turicamycin-inhibitor of N-glycosylation of proteins on expression of (?1 integrin,FAK and cyclin D1 in high glucose-induced glomerular mesangial cells (CMC), and to explore the mechanism concerned. Methods Cultured HBZY-1 rat mesangial cells were divided into 5 groups:control group;high glucose group;mannitol group;high glucose plus TM group; TM group.The expression of ?1 integrin was measured by flow cytometry, expression of FAK and cyclin D1 was measured by immunohistochemistry,and proliferation of GMCs was measured by MTT. Results There was a little expression of ?1 integrin, FAK and cyclin D1 on normal mesangial cells. High glucose induced the proliferation and increased the expression of ?1 integrin and cyclin D1.There was no significant difference in mannitol group as compared to control group.The expression of (?1 integrin FAK and cyclin D1 decreased notably by TM.TM could also decrease proliferative abilitiy of cells. All the effects of TM were dose-dependent. Conclusion Through blocking glycosylation of glycoprotein, TM can suppress cellular proliferation and expression of pi integrin induced by high glucose in a dose-dependent manners,then pi integrin affects the expression of FAK and cyclin D1.

Objective:To study the influence of protein transduction domain (PTD)-oligomerization domain (OD)-HA fusion proteins on apoptosis of bcr/abl-positive cell lines. Methods:bcr/abl-positive cells were treated with PTD-OD-HA protein. The apoptoses of the cells were detected by flow cytometry (FCM), DNA ladder and transmission electron microscopy (TEM), and the levels of apoptosis-related genes bax and bcl-2 were detected by RT-PCR and Western blotting. Results:FCM examination demonstrated that PTD-OD-HA protein induced the apoptosis of bcr/abl-positive cells; DNA ladder showed that the classic DNA ladders appeared in BaF3-P210 and K562 cells after 48 h treatment with PTD-OD-HA proteins; the apoptoses of BaF3-P210 cells were observed by TEM; the levels of bax in mRNA and protein increased in BaF3-P210 and K562 cells, and bcl-2 decreased. Conclusion:PTD-OD-HA proteins specifically induced the apoptosis of bcr/abl positive cells.

Objective To investigate the response of mesenchymal stem cells in mice to mediumdose X-ray irradiation in vitro.Methods The mouse mesenchymal stem cell line C3H10T1/2 was submitted to 3.5 Gy X-ray irradiation.Hoechst33258 staining of adherent cells and Annexin V-FITC staining and flow cytometry analysis of suspension cells were performed respectively to assess cellular apoptosis at 3,6,12,24,48,72 h and 1 week after irradiation.SA-β-gal staining was performed to analyze the cellular senescence at 24,48,72 h and 1 week after irradiation.The mRNA level of both Fas with its ligand FasL and p53 with its downstream target p21 WAF1 were measured by Real-Time PCR analysis.The expression of Fas protein was determined by immunofluorescence staining.Results An increased apoptosis was observed at 3 h after irradiation with apoptosis rate 11.72％ ± 1.61％ ( t =9.01,P ＜0.01 ),the apoptosis rate reached the peak level at 12 h 20.52％ ± 1.96％ (t =16.27,P ＜ 0.01 ),and then declined progressively to normal level at 48 h 4.93％ ±0.46％ (t =2.26,P ＞0.05).The SA-β-gal positive rate of post-radiation cells at 72 h was 53.33％ ± 5.62％,significantly higher than that of normal control 3.24％ ± 0.39％ (t =17.77,P ＜ 0.01 ).The level of Fas,FasL mRNA was found to be elevated 3 h after irradiation with a peak at 12 h,and no differences were found l week later.The level of Fas protein was observed to reach the peak at 12 h after irradiation.The occurrence of peak level of Fas/FasL mRNA and protein was consistent with that of apoptosis of C3H10T1/2 cell.A transient up-regulation of p53,p21 WAF1 mRNA expression was found at 12 h after irradiation followed by a significant increase later at 72 h after irradiation.The occurrence of the two peaks of p53,p21WAF1 mRNA expression were coincident with that of cellular apoptosis and senescence,respectively.The levels of p53,p21WAF1 mRNA in senescence group were significantly higher than those of apoptosis group ( t =17.85,13.36,P ＜ 0.01 ).Conclusions The MSC cell line C3H10T1/2 was sensitive to medium-dose X-ray irradiation.Cell apoptosis occurred immediately after irradiation and cellular senescence happened at advanced stage.Both Fas/FasL and p53/p21 WAF1 signal pathway mediate the injury of C3H10T1/2 cell to medium-dose X-ray irradiation exposure.

Objective To investigate the relationship between the "prominent laterality of the posterior cerebral artery (PLPCA)" found on magnetic resonance angiography (MCA) and the size and distribution of cerebral infarction and the National Institutes of Health Stroke Scale (NIHSS)scores in patients with occlusion of the M1 segment of the middle cerebral artery (MCA).Methods Fifty patients with acute cerebral infarction caused by the occlusion of the M1 segment of MCA were divided into PLPCA positive group (n =24) and PLPCA negative group (n =26) according to MRA manifestation.the NIHSS scores,size of cerebral infarction scores,and constituent ratios of distribution in all the feeding subregions of MCA in both groups were compared.Results The proportions of the patients with ≥3 risk factors (9/24 vs.18/26,P =0.046),NIHSS scores (5.4 4.4 vs.10.4 ±4.9,t = -3.690,P =0.001),and the size of cerebral infarction scores (1.92 ± 1.10vs.2.88 ± 1.37,t = -3.690,P =0.001) in the PLPCA positive group were significantly lower than those in the PLPCA negative group.The proportions of the patients with cerebral infarction involying the middle branch of the MCA territory (6/24 vs.19/26,P =0.002) and the posterior branch of the MCA territory (2/24 vs.5/26,P ＜0.001) in the PLPCA positive group were significantly lower than those in the PLPCA negative group.The proportions of the patients whose infarction involving the area of the posterior watershed zone were significantly higher than those in the PLPCA negative group (6/24 vs.1/26,P =0.045),and the proportions of complete infarction were significantly lower than those in the PLPCA negative group (0/24 vs.6/26,P =0.023).Conclusions When MCA M1segment was occluded,if PLPCA were observed on MRA,it indicated that the infarct size was smaller and the NIHSS score was lower.The infarction was less involved in the middle and post branches of MCA,and it is prone to have posterior watershed infarction.

Good self-management behaviors can control symptoms of the patients with osteoarthritis, improve the patients' joint function and quality of life. Patients' self-management behaviors have been impacted by disease knowledge, self-effcacy, emotional state, and social support. All the above factors should been taken into full consideration when intervening. Self-management program is an intervention mode which can improve patient self-management behaviors and promote patient health.

Objective To construct a recombinant eukaryotic expression plasmid of glycosylphosphatidyl inositol(GPI)-anchored bcr/abl and explore its expression at mRNA and protein level.Methods The gene fragment encoding bcr/abl was amplified by PCR using the plasmid containing the cDNA sequence of P210 as template and then inserted into a eukaryotic expression vector pBudCE4.1.The constructed recombinant plasmid pBudCE4.1-bcr/abl was identified by restriction analysis and DNA sequencing.Lymphocytes were isolated from human peripheral blood and their total RNA was extracted.The gene fragment encoding GPI was amplified by RT-PCR using the obtained RNA as template and was inserted into the constructed recombinant plasmid pBudCE4.1-bcr/abl in order to anchor GPI and bcr/abl.The constructed recombinant plasmid pBudCE4.1-bcr/abl-GPI was transfected into COS-7 cells,and the expressions of objective fragment were detected by RT-PCR and Western blotting.Results The results of restriction analysis,PCR and DNA sequencing proved that GPI-anchored bcr/abl fusion fragment was correctly inserted into vector pBudCE4.1.The expression of bcr/abl fusion gene and fusion protein were identified in transfected COS-7 cells and on their membrane.Conclusion The recombinant plasmid pBudCE4.1-bcr/abl-GPI was successfully constructed and expressed on the membrane of COS-7 cells,which found a basis of cell immunity with GPI-anchored bcr/abl fusion gene.

Baizi Yangxin tablets are common Chinese medicine used for the treatment of heart palpitations, insomnia and irritable forgetful. To evaluate the total amount of heavy metals evaluation, artificial gastric juice was used as juice samples and the extracted soluble heavy metals were extracted by microwave digestion technology. An analytical method of bionic extraction microwave digestion with inductively coupled plasma mass spectrometry ( ICP-MS ) was established for the determination of trace metals, such as Co, Cr, Cu, Ba, Cd, Mn, Ni, Pb, Hg, Sr and Zn, in 18 batches of Baizi Yangxin tablets. The correlation coefficient of linear regression equation for different elements ranged from 0. 9989 to 1. 0000, the detection limit was 0. 19-5. 6 μg / L, and the repeatability of the method was less than 6. 2% , the precision of the RSD value was less than 5. 6% , and the recovery rate was 87. 7% -101. 9% . According to “the standard of the heavy metal in the standard of import and export of medicinal plants and preparations”, the contents of Cd, Cu and Pb in the 18 batches were not exceed the standard, but the Hg content (7. 68 mg / kg) exceeded the standard value. The bionic extraction-ICP-MS method provide a reference basis for the safety of the proprietary Chinese medicine's study.

OBJECTIVETo investigate the expression of HERC4 in human breast cancer tissues and its relationship with the clinicopathological features.

METHODSRT-qPCR was used to detect mRNA expression of HERC4, and Western blotting and immunohistochemistry were employed to detect protein expression of HERC4 in 67 breast cancer tissues and adjacent breast tissues.

RESULTSThe results of RT-qPCR showed a significantly higher mRNA expression of HERC4 in breast cancer tissues than in the adjacent breast tissues (P<0.05). Western blotting demonstrated HERC4 over-expression in breast cancer tissues compared with the adjacent breast tissues (P<0.05). Immunohistochemistry revealed HERC4 expression located predominantly in the cell cytoplasm. Positive HERC4 expression was detected in 61.2% of the breast cancer tissues as compared to 19.4% in the adjacent breast tissues, and its expression level was closely correlated with TNM stage and the histological grade (P<0.05).

CONCLUSIONHERC4 is correlated with the tumorigenesis and progression of breast cancer and may serve as a potential biomarker for early diagnosis and also as a potential therapeutic target in breast cancer.

Blotting, Western ; Breast ; metabolism ; Breast Neoplasms ; metabolism ; Disease Progression ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Real-Time Polymerase Chain Reaction ; Ubiquitin-Protein Ligases ; metabolism

Objective To analyze the knowledge and perceptions of brucellosis and human behavior in different populations in Jingyuan County Gansu Province,and to provide a scientific basis for carrying out health education and prevention accurately on brucellosis.Methods In 2015,using two-stage cluster sampling method,nine towns with more accumulative incidence of brucellosis in the past 3 years in Jingyuan Country were selected,5 villages were selected from each town,occupational exposure and patients with brucellosis were selected in each village as respondents.A questionnaire survey was carried out to collect basic information,brucellosis related knowledge,population's behavio.Results The overall awareness rate of prevention and control knowledge on brucellosis was 44.10％ (12 943/29 348),included 809 people with the occupation exposure population and 203 patients with brucellosis,among them,the rate of patients with brucellosis knowledge was 50.40％ (2 967/5 887),occupational exposure population was 42.52％ (9 976/23 461);There were significant differences in the awareness rate of knowledge on prevention and control of brucellosis in population of different gender,age,education and years of work experience (x2 =84.413,166.100,207.200,16.822,P ＜ 0.01);of the following parameters:shared water,peel dead lamb,How to deal with flow products,treatment of abortion without gloves,masks,not wearing gloves,masks when lambing ,to give livestock vaccines and drugs,eat dead cow,lamb,and sale of diseased,dead livestock,slaughter livestock,there were statistical significant differences between patients with brucellosis and exposure people (x2 =13.940,27.965,30.031,19.575,22.597,21.139,14.524,436.450,8.482,P ＜ 0.05).Conclusions The occupational exposure population has a low knowledge awareness rate;high risk behaviors have higher risk of brucellosis infection in Jingyuan County.We should carry out health education and high-risk behavior intervention in targeted population.

Objective To construct double expression retroviral vectors targeting chronic myeloid leukemia (CML) b3a2-type mRNA and investigate its effect on the phenotype of K562 cells. Methods The eGFP coding sequence was inserted into the retroviral vector pMSCV-neo to construct pMSCV/GFP, then H1-RNA pol III-based transcription cassettes was subcloned into it to form pMSCV/GFP-H1-BCR/ABL40AS. Two control vectors pMSCV/GFP-H1-BCR/ABL40S & pMSCV/GFP-H1-BCR/ABL80AS were constructed in addition. All these constructions were identified by restriction enzyme analysis and DNA sequencing. After that, the recombinant vectors were transferred into retrovirus packaging cell line PT67 by using lipofectamine2000, and G418 were used to select stable virus-producing cell lines. Viral titer was determined by infection of NIH3T3 cells sequentially. The cell-growth curve was assayed, cell apoptosis was checked with Annexin V-PE/7AAD double staining and flow cytometry analysis after 24-hour infection, the PKR phosphorylation was assayed by Western blotting. Results The plasmids were successfully constructed. Four cell lines, named as PT67-MSCV/GFP, PT67-40as, PT67-40s and PT67-80as were gained by G418 selection, and virus titers were 6.2?10~ 5 , 5.6?10~ 5 , 4.6?10~ 5 and 6.0?10~ 5 CFU/ml respectively. PT67-40as suspensions could induce K562 cell apoptosis by (22.54?3.19)%, significantly different from PT67-MSCV/GFP or PT67-40s (P