Objective:To study the ethical attitude of infertility patients and the public towards the artificial insemination technology,with the aim of its better clinical application.Methods:A questionnaire among 220 infertility patients and 720 persons of different social walks,followed by a contrastive study of the results.Results:There is distinctive difference(P

We studied the function of connexin 43 (Cx43) gene in Xiao Meishan swine bone marrow mesenchymal stem cells (BMSCs) resulting from overexpression. Cx43 eukaryotic expression vector (pEGFP-Cx43) was constructed and transfected into BMSCs by nucleofector, after detecting the transfection efficiency; the expression of Cx43 was verified by RT-PCR, immunofluorescence and western blotting. Furthermore, we detected its cell cycle and apoptosis through flow cytometry. Our results show that pEGFP-Cx43 plasmid was successfully constructed, and green fluorescence in pEGFP-Cx43 transfected BMSCs was highly expressed with 60% transfection efficiency. In transgenic Xiao Meishan swines BMSCs, the expression level of Cx43 mRNA and protein were up-regulated. Meanwhile, the ability of cell proliferation was significantly increased, and the apoptosis rate was significantly reduced. Taken together, Cx43 overexpression could promote the proliferation of Xiao Meishan swine's BMSCs and markedly reduce their apoptosis, which provides evidence for in vivo research.
Animals ; Animals, Genetically Modified ; Apoptosis ; Cell Proliferation ; Connexin 43 ; metabolism ; Flow Cytometry ; Genetic Vectors ; Mesenchymal Stromal Cells ; metabolism ; Plasmids ; Swine ; genetics ; Transfection

Objectives To explore intraocular drug concentration changes and the pharmacokinetics after topically applied of bevacizumab with armexin A5-associated liposome on rabbit eyes.Methods A total of 105 healthy New Zealand white rabbits were selected and divided randomly into 3 groups (group A,B and C),and each group had 35 rats.Bevacizumab with annexin A5-associated liposome,bevacizumab liposome and bevacizumab were topically applied 50 μl respectively on right eyes of rabbits in group A,B and C,respectively.Aqueous,vitreous body and retina/choroid were obtained at 5,15,30 minutes and 1,2,4,8 hours and the free bevacizumab concentrations in these ocular tissues were measured by ELISA (enzyme linked immunosorbent assay).DAS 2.1.1 software was used to fit the pharmacokinetic parameters.Results The peak drug concentrations in aqueous humor of the eyes in group A,B,C were at 15 minutes after topical administration and the difference was statistically significant (F=301.061,P＜ 0.01).The peak drug concentrations in vitreous of the eyes in the group A,B,C were at 2 hours after topical administration and the difference was statistically significant (F=885.997,P＜ 0.01).The peak drug concentrations in retina/choroid of the eyes in the group A,B,C were at 1 hour after topical administration and the difference was statistically significant (F=644.908,P＜0.01).Least significant difference pair-wise test found that the drug concentrations in aqueous humor,vitreous and retina/choroid of group A was higher than that of the group B and C respectively (P＜ 0.05),while that of the group B and C had no significant different (P＞ 0.05).Pharmacokinetic fitting analysis found that the half-life (t1/2) of bevacizumab in aqueous humor were 1.14,1.29,1.29 hours,the distribution t1/2 were 1.40,1.50,1.42 hours and the eliminated t1/2 were 2.62,2.84,2.73 hours in vitreous,the distribution t1/2 were 2.61,2.99,2.70hours and the eliminated t％ were 2.61,2.99,2.70 hours in retina/choroid respectively for the 3 groups.Changes of bevacizumab concentration in aqueous humor of rabbit eyes for 3 groups was complied with one compartment model,and that in vitreous body and retina/choroid complied with two compartment model.Conclusions Topically applied annexin A5-associated liposome has higher ocular concentrations of bevacizumab than those of controls.Changes of bevacizumab concentration in aqueous humor of rabbit eyes was complied with one compartment model,and that in vitreous body and retina/choroid complied with two compartment model.

ObjectiveTo establish a method to induce heterotopic ossification (HO) after spinal cord injury. Methods10 New Zealand White rabbits underwent a complete transection of the thoracic (T10) spinal cord. Just after transection, the right hind limb of every rabbit was immobilized with a plaster support to keep the knee in extension and the hip unrestricted. The plaster was temporarily removed once a day (six times per week) to allow the knees passively mobilized at the maximum range for 5 min. Local swelling, local skin temperature and grade of ossification of the paraplegic limbs were observed. After 5 weeks, the rabbits were sacrificed, and the quadriceps muscles of the right hind limbs were removed for the histological examination. ResultsAfter 5-week immobilization and temporary passive mobilization, HO was successfully induced in all 10 rabbits. There was local skin temperature increase and local swelling in the right hind limbs. The histological changes were similar to those observed in clinical HO. Conclusion5-week immobilization and temporary passive mobilization approach may successfully induce HO, and it may be used to study the pathogenesis of HO after spinal cord injury.

Formative evaluation is a medical education evaluation form used to correct its own track in the course of activities so as to get better effect. In the medical education, it can significantly improve the medical students' ability such as active learning ability, relearning ability, ability to master and apply know l edge, ability of scientific thinking and debating, ability of innovation and analysis, ability of association and communication, and so on.

To monitor the trans-differentiation from adult stem cells to germ cells, we analyzed the vasa expression of goat testicular tissues in different ages and constructed the germ cell specific reporting vector pVASA-EGFP. The expression of vasa was verified by RT-PCR and immunofluorescence. The vector pVASA-EGFP was constructed by molecular technology, then transfected into goat bone mesenchymal stem cells (BMSCs) by Lipofectamine 2000. Moreover, we observed the expression of the vector through green fluorescent protein (GFP). Immunofluorescence results show that Vasa was expressed in all groups of goat testicular tissues, RT-PCR results show that the levels of vasa mRNA in 3-month group and 10-month group were significantly higher than that in 10-day group. Sequencing and restriction enzyme results show that the vector was successfully constructed. After transfection and RA treatment, GFP expression was observed, which proved the validity of our reporting system. All the results proved that vasa was expressed in different ages in goat testicular tissues, and the vector pVASA-EGFP is efficient in monitoring the trans-differentiation in vitro, which paves the way for further characterization and screening of the trans-differentiation of goat BMSCs.
Animals ; Cell Transdifferentiation ; Genes, Reporter ; Genetic Vectors ; Germ Cells ; cytology ; Goats ; Green Fluorescent Proteins ; Male ; Mesenchymal Stromal Cells ; RNA, Messenger ; Testis ; metabolism ; Transfection

Hot Temperature ; Humans ; Industry ; Occupational Exposure ; Occupational Health

This study was to explore a better three-dimensional (3-D) culture method of chondrocyte. The interpenetrating network (IPN) gel beads were developed through a photo-cross linking reaction with mixed barium ions and calcium ions at the ratio of 5:5 with the methacrylic alginate (MA), which was a chemically conjugated alginate with methacrylic groups. The second generation of primary cartilage cells was encapsulated in the MA gel beads for three weeks. In the designated timing, HE stain, Alamar blue method and Scanning electron microscopic were used to determine the cartilage cells growth, proliferation and the cell distribution in the scaffolds, respectively. The expression of type II collagen was investigated by an immunohistochemistry assay and the glycosaminoglycan content was quantitatively evaluated with the spectrophotometry of 1, 9 dimethylene blue assay. Compared to the alginate control group, the deposition of glycosaminoglycan was significantly upregulated in IPN-MA gel beads with higher cell proliferation. The secretion of extracellular matrix and proliferation of chondrocyte in methacrylic alginate gel beads were higher than that in Alginate beads. Cells were able to attach, to grow well on the scaffolds under scanning electron microscopy. The result of immunohistochemistry staining of collagen type II was positive, confirming the maintenance of chondrocyte phenotype in methacrylic alginate gel beads. This study shows a great potential for three-dimensional culture of cartilage.
Alginates ; chemistry ; Barium ; chemistry ; Calcium ; chemistry ; Cartilage ; cytology ; Cations ; Cell Culture Techniques ; instrumentation ; Cells, Cultured ; Chondrocytes ; cytology ; Collagen Type II ; chemistry ; Glucuronic Acid ; chemistry ; Glycosaminoglycans ; chemistry ; Hexuronic Acids ; chemistry ; Metals ; chemistry ; Microscopy, Electron, Scanning

Objective To explore a new peptide-based approach independent of HLA to generate antigen-specific CD+ CD8+T cells. Methods Peripheral blood mononuclear cells(PBMC) were stimula- ted for 6 h with IE-1 peptide pool. Then the activated IFN-γsecreting ceils were tested by immunomagnetic selection. And the selected cells were cultured with radio-inactivated PBMC in medium with 100 IU/ml IL-2 for 4 weeks. Results The generated T cell lines consisted of IE-1 specific CD4+ T (6.88%) and CD8+ T cells 92.99%, which demonstrated antigen-specific killing and cytokine secretion. Conclusion T ceils can be proliferated with this new procedure, and maintain its phenotype and antigen-specific function.

Aim To investigate the effects of valsartan on activation of signal transducers and activators of transcription 1,3 in glomerular mesangial cells(GMCs) under high concentration of glucose.Methods we used high concentration glucose and valsartan to stimulate the cultured rat GMCs in vitro.The protein expressions of signal transducer and activatior of transcription 1,3(STAT1,STAT3),p-STAT1 and p-STAT3 were observed by Western blot.The protein synthesis of TGF-?_1,fibronectin and type IV collagen in the supernatants of the GMCs were detected by enzyme-linked immunoadsorbent assay(ELISA) and radioimmunoassay.TGF-?_1 mRNA was measured by reverse transcription and polymerase chain reaction(RT-PCR).Results Compared with low glucose control group,the expressions of p-STAT1,p-STAT3 and TGF?_1 mRNA were significantly increased in GMCs under high concentration glucose medium and there observed the high concentration of TGF-?_1,fibronectin and type IV collagen in the supernatants.The expression levels of p-STAT1,p-STAT3 and TGF-?_1 mRNA were significantly lower in the valsartan group than in those the high concentration glucose group.The concentration of TGF-?_1,fibronectin and type IV collagen in the supernatants in the valsartan group were lower than that in the high concentration glucose control group.Conclusion Valsartan can inhibit overproduction of TGF-?_1 and ECM proteins in GMCs under high concentration of glucose,partly by regulating the phosphorylation of STAT1 and STAT3.