Genetically engineered mice is one of the powerful research systems in exploring the gene protein life phenomenon,which is useful in generating the laboratory animal model of human diseases and developing the new drugs. With the development of functional genomics and model organism in life science area, biological medicine research is in need of genetically engineered mice deadly. There are still some problems to be resolved, including death effect, no or low expression and no phenotype. It is essential to improve the basic research related to genetically engineered mice.

A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.

A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.

Objective To evaluate the correlation between serum uric acid with cognitive disorder after acute cere?bral infarction by prospective study. Methods Four hundred consecutively enrolled patients of acute cerebral infarction were divided into no cognitive impairment group and cognitive impairment group according to the assess of Montreal Cog?nitive Assessment (MoCA). Univariate analysises were conducted in the potential risk factors of cognitive impairment in?cluding age, sex, smoking, alcohol, hypertension, diabetes, dyslipidemia, level of education, infarction in key parts, atrial fibrillation, serum uric acid, blood homocysteine between two groups. The statistically significant indicators in univariate analysises were used as independent variables and the scores of MoCA were used as the dependent variable to conduct multiple linear regression analysis. The assessment on the risk of cognitive impairment after cerebral infarction were con?ducted according to serum uric acid, sex, age and TOAST classification further. Results Serum uric acid was indepen?dent risk factors of cognitive disorder after acute cerebral infarction. The risk of cognitive disorder after acute cerebral in?farction was significantly increased in patients with high level of serum uric acid than with normal level and the relative risk was 1.35,95%CI(1.098,1.660). Especially for the young, male or patients with cerebral infarction in classification of small artery occlusion, the risk increased further, and the relative risk was 1.513, 95%CI(1.092, 2.096)1.412, 95%CI (1.125, 1.771)and 1.464, 95%CI(1.128, 1.900)respectively. Conclusion Exaltation of Serum uric acid was indepen?dent risk factor of cognitive disorder after acute cerebral infarction. The risk of cognitive disorder after acute cerebral in?farction was significantly increased in patients with high level of serum uric acid than with normal level, and especially for the young, male and patients with cerebral infarction in classification of small artery occlusion, the risk increased fur?ther.

Objective: To observe the effect of overexpression of miRNA200a (miR-200a) recombinant lentivirus on the expression of Wnt/β-catenin signaling pathway in mouse lung epithelial cell line MLE-12 induced by silica (SiO₂) . Methods: The mice were divided into SiO₂ control group (SiO₂) , virus control group (SiO₂+Lv-NC) group and overexpressing miR-200a virus group (SiO₂+Lv-miR-200a). The expression of β-catenin, MMP2, MMP9, TCF-4 and Cyclin D1 mRNA and protein were detected by realtime-PCR and western blot after incubating cells for 18 h stimulating at the final concentration of 200 μg/ml of SiO₂. Results: The expression of miR-200a in MLE-12 cells of SiO₂+Lv-miR-200 a group was significantly higher than that in SiO₂ group and SiO₂+Lv-NC group. The mRNA and protein expression of β-catenin, MMP2, MMP9, TCF-4 and Cyclin D1 in MLE-12 cells of SiO₂+Lv-miR-200a group were significantly lower than those in SiO₂ group and SiO₂+Lv-NC group (P<0.05) . Conclusion Overexpression of miR-200a can inhibit the expression of related genes of Wnt/β-catenin signaling pathway in silica-induced mouse lung epithelial cells.

OBJECTIVETo investigate the prophylactic and therapeutic effect of oxymatrine on experimental liver fibrosis and to reveal its mechanism.

METHODSBy establishing D-galactosamine-induced rat liver fibrosis model, we observed the effect of oxymatrine on serum and tissue biochemical indexes, content of liver hydroxyline, expression of TGF?1 mRNA and changes of tissue pathology.

RESULTSThere was a decline of liver hydroxyline and serum AST and ALT in oxymatrine group compared to those of the D-GalN group. The hydroxyline content in oxymatrine pretreatment group was (0.50 0.11)mug/mg compared with (0.99 0.14)mug/mg in D-GalN group (t=8.366, P<0.01). The content in oxymatrine treatment group was (0.44 0.04)mug/mg compared with 0.70 0.06 in D-GalN group (t=9.839, P<0.01). The SOD activity was (149.81 15.28) NU/mg in oxymatrine pretreatment group and (95.22 16.33) NU/mg in the model group (t=7.309, P<0.01); (157.68 19.54) NU/mg in the treatment group compared with (119.88 14.94) NU/mg in the model group (t=4.348, P<0.01). MDA in the pretreatment group was (2.06 0.17) nmol/mg, lower than (4.57 0.37) nmol/mg in the model group (t=17.529, P<0.01). In the treatment group, it was (1.76 0.24)nmol/mg, lower than (3.10 0.17) nmol/mg in the model group (t=12.697, P<0.01). TGF?1 mRNA reduced in the pretreatment and treatment groups as compared with that in the model group (0.21 0.01 vs 0.50 0.01, t=48.665, P<0.01; 0.18 0.02 vs 0.38 0.01, t=22.464, P<0.01). Electron microscopy showed that oxymatrine group had milder hepatocyte degeneration and less fibrosis accumulation than did the model group. Microscopy revealed wide septa expansion from the portal area to the central venous, piecemeal and confluent necrosis and pseudo-nodular formation in part of the lobular in the model group. While in oxymatrine group these lesions were much improved.

CONCLUSIONSOxymatrine shows prophylactic and therapeutic effect in D-galactosamine induced rat liver fibrosis. This is partly by protecting hepatocyte and suppressing fibrosis accumulation through anti-lipoperoxidation.

Alkaloids ; therapeutic use ; Animals ; Anti-Arrhythmia Agents ; therapeutic use ; Calcium Hydroxide ; metabolism ; Chemoprevention ; Disease Models, Animal ; Galactosamine ; Liver Cirrhosis ; chemically induced ; drug therapy ; metabolism ; pathology ; prevention & control ; Liver Function Tests ; Male ; Quinolizines ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism ; Transforming Growth Factor beta ; genetics ; metabolism

Objective:To study the different factors affecting platelet production post transplantation of hematopoietic stem cells (HSCs) isolated from different sources in order to explore novel options for treating platelet depletion following HSCs transplantation.Methods:HSCs and their downstream derivatives including myeloid and lymphoid cells (i.e., collective of mononuclear cells (MNCs)), were isolated from E14.5 fetal liver (FL) and bone marrow (BM) of 8-week-old mice by Ficoll separation technique. These cells were subsequently transplanted into the tibia bone marrow cavity of recipient mice post lethal myeloablative treatment in order to construct the FL-MNCs and BM-MNCs transplantation mouse model. Routine blood indices were examined in these recipient mice. The chimeric rate of donor cells in recipient peripheral blood cells were determined by flow cytometry. Different groups of cells involved in platelet reconstruction were analyzed. CD41 +megakaryocytes were sorted from fetal liver or bone marrow using magnetic beads, which were then induced to differentiate into platelets in an in vitro assay . Quantitative RT-PCR was used to detect the expression of platelet-related genes in CD41 +megakaryocytes from the two sources. Results:Both the FL-MNCs and the BM-MNCs transplantation groups resumed normal hematopoiesis at the 4th week after transplantation, and the blood cells of the recipient mice were largely replaced by the donor cells. Compared with the mice transplanted with BM-MNCs, the platelet level of mice transplanted with FL-MNCs recovered faster and were maintained at a higher level. At week 4, the PLT level of the FL-MNCs group was (1.45±0.37)×10 12/L, and of the BM-MNCs group was (1.22±0.24)×10 12/L, P<0.05. The FL-MNCs contain a higher proportion of hematopoietic stem cells (Lin -Sca-1 +c-Kit +)(7.60%±1.40%) compared to the BM-MNCs (1.10%±0.46%), P<0.01; the proportion of the megakaryocyte progenitor cells (Lin -Sca-1 -c-Kit +CD41 +CD150 +) and mature megakaryocyte cells (CD41 +CD42b +), also differ significantly between the FL-MNCs (3.05%±0.22%, 1.60%±0.06%, respectively) and the BM-MNCs (0.15%±0.02%, 0.87%±0.11%, respectively) groups, both P<0.01. In vitro functional studies showed that FL-MNCs-CD41 +megakaryocytes could produce proplatelet-like cells more quickly after induction, with proplatelet-like cells formation on day 3 and significant platelet-like particle formation on day 5, in contrast to bone marrow-derived BM-MNCs-CD41 +megakaryocytes that failed to form proplatelet-like cell on day 5. In addition, FL-MNCs-CD41 +cells expressed higher levels of platelet-related genes, Mpl (3.25-fold), Fog1 (3-fold), and Gata1 (1.5-fold) ( P<0.05). Conclusion:Compared with the BM-MNCs group, the FL-MNCs transplantation group appears to have a more efficient platelet implantation effect in the HSCs transplantation recipient in vivo , as well as a higher platelet differentiation rate in vitro. This might be related to a higher proportion of megakaryocytes and higher expression levels of genes such as Mpl, Fog1, and Gata1 that could be important for platelet formation in FL-MNCs-CD41 +cells. Further exploration of the specific functions of these genes and the characteristics of the different proportions of the donor cells will provide valuable clues for the future treatment of platelets reconstitution after HSCs transplantation clinically.

OBJECTIVE: To investigate occupational stress and its influencing factors in workers of electronic manufacturing.METHODS: The judgment sampling method was used to select 2 251 workers as study subjects. The Simple Occupational Stress Questionnaire and the Effort-Reward Imbalance( ERI) Questionnaire were used to investigate job demand-control(JDC) and ERI occupational stress respectively. RESULTS: Among the 2 251 workers,the prevalence rates of high occupational stress in JDC and ERI models were 56. 4% and 10. 1%,respectively. The positive rate of JDC occupational stress was higher than that of ERI( P < 0. 01). The multivariate logistic analysis indicated that workers with assembly line had higher risk of JDC and ERI occupational stress than workers without assembly line( P < 0. 05). Workers with labor dispatching had higher risk of JDC and ERI occupational stress than workers with long-term contract( P < 0. 05). Workers exposed to occupational hazards had higher risk of JDC and ERI occupational stress than workers not exposed to occupational hazards( P < 0. 05). Workers with disease had higher risk of JDC and ERI occupational stress than healthy workers( P < 0. 05). CONCLUSION: JDC is the main occupational stress model in workers of electronics manufacturing factory. The main influencing factors are assembly line,labor dispatching,weekly work time,exposure to occupational hazards and illness.

Objective: To explore the role of serum microRNA (miRNAs) levels in the detection of pneumoconiosis, and to establish a combined application model of multiple serums miRNAs for pneumoconiosis diagnosis. Methods: 152 cases were selected in the case group and the control group respectively. The TaqMan Low Density Array method was used to screen out the candidate miRNAs for early screening of pneumoconiosis, and RT-qPCR was used to verify. According to the area under the curve (AUC) , the sensitivity and specificity of the candidate indicators were investigated. The logistic regression model was established by the two-class logistic regression model. Results: The expression of 7 candidate miRNAs in the serum of pneumoconiosis patients was significantly different (P<0.05) . The receiver operating curve (ROC) of the above 7 miRNAs was analyzed, miRNA-21, miRNA-200c, miRNA-16, miRNA-206, miRNA-155, miRNA-29a had statistical significance, and their ROC-AUC is 0.629~0.932. Logistic regression model was: logitP=13.769+0.536×miRNA-21-0.878×miRNA-200C-0.012×miRNA-16-0.111×miRNA-206+0.117×miRNA-155-1.192×miRNA-29a. Conclusion Multiple serum miRNAs combined application models may be used for the diagnosis of pneumoconiosis patients.