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

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.

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

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.

Objective To evaluate the effect of lentinan polysaccharide (LPS) on Alzheimer's disease (AD) rats, and providing a new idea for prevention and treatment of Alzheimer's disease (AD).Methods 96 SPF rats were randomly divided into control group, model group, LPS low dose-treated group, LPS middle dose-treated group, LPS high dose-treated group and positive control group(piracetam treated), each had 16 rats.The model of AD was induced by injection of Aβ25-35(80 pmol/μL), rats in four treatment groups were treated with LPS (200 mg/kg), LPS (400 mg/kg), LPS (600 mg/kg) and piracetam(600 mg/kg) orally once a day for 60 days while control group and model group were given equal-dose saline.After treatment, the ability of navigation and memory were evaluated by water maze test (MWM), and then rats in all groups were sacrificed to obtain the hippocampus.HE staining was used to observe the histopathological changes, the expressions of TACE,Aβ,BACE1 and PP2A protein in hippocampus were detected by Western blot.Results Compared with the model group, high dose LPS could significantly improve the ability of learning and memory (P<0.01) and improve the ability of locating navigation (P<0.01).HE staining results showed that LPS treatment could repair the rat hippocampus injury induced by Aβ25-35 .Western blot analysis showed that the levels of Aβ, and BACE1 were down-regulated in LPS group compared with the model group, while the protein level of PP2A and TACE was up-regulated.Conclusion LPS has a protective effect on Aβ25 -35 induced AD model, and it is expected to be a strategy for treatment of AD .

Objective To investigate the proliferation, immune phenotype and cytotoxicity on different cell lines of cytokine-induced killer (CIK) cells collected from healthy donors. Methods Peripheral blood mononuclear cells (PBMC) from healthy donors were induced to become CIK cells by adding cytokines including rhIL-2, rhIFN-γand CD3 McAb. The proliferation of CIK cells was tested by blood cell recording board. The CIK cells were analyzed on different time points by FACS. The cytotoxicity of CIK cells against different tumor cell lines, such as K562, BJAB, A549, MCF-7 and HepG2, was detected by MTT assays on day 13. Results CIK cells quickly proliferated from day 5, and expanded by 182-fold after 20-day culture. The immunophenotypes of CD3+, CD3+CD8+and CD3+CD56+were (97.83±1.03)%, (77.12±1.60)%and (27.58± 2.02)%. The percentages of CD3+, CD3+CD8+and CD3+CD56+increased noticeably (P＜0.01). According to the effector-tar-get ratio of 40∶1, the activity of CIK cells against tumor cells K562, BJAB, A549, MCF-7 and HepG2 were (88.89±7.22)%, (75.42±9.52)%, (63.19±5.67)%, (43.53±5.67)%and (42.63±7.69)%. The experiments showed that CIK cells possessed high-er antitumor cytotoxic activity. Conclusion CIK cells can be largely capacity cultured by adding cytokines in vitro. CIK cells were a highly efficient cytotoxic cell against tumors, and had clinical application potentials.

Objective To explore the anti-inflammatory mechanisms of three stains. Methods Airpouch induced acute inflammation rat model was developed by subcutaneous injection of carrageenan.Inflammatory invasion and tumor necrosis fator-alpha (TNF-α) expression levels of the inflammed skins were examined by immunohistochemical methods and digital image analysis. The anti-inflammatory action of the 3 different statins was compared. Results Stains could inhibit inflammatory invasion and the expression of TNF-α. Conclusion The statins have demonstrated strong inhibition of inflammation.

BACKGROUND:The finite element analysis method is more accurate and fast to construct the three-dimensional model of the human skeleton and design the bone surgical medical instrument.
OBJECTIVE:To establish locking plate model according to the clavicle model, analyze and evaluate stress distribution of locking plate of the finite element model under bending and torsion conditions.
METHODS:Chest scan was carried out in a healthy young adult male by adopting 64-row spiral CT and his two-dimensional image data were gotten. The obtained data were analyzed with Mimics 10.0 software to establish the three-dimensional clavicle finite element model. The clavicle locking fixation plate model was established by applying the UG software. The locking fixation plate was evaluated by utilizing the abaqus software when the plate was bent while down to give force of 200 N, and twisted while 200 N?mm, to simulate the force and analyze the stress distribution of the locking plate.
RESULTS AND CONCLUSION:Based on the original image parameters provided by CT, this experiment produced a three-dimensional model of the clavical titanium plate which fitted better to bones. This model can obtain a single individual, personalized plate by three-dimensional printing technology. The finite element analysis basical y can simulate the actual stress of the plate. For straight plate and“S”-shape plate, in lateral bending and axial torsion loads, the maximum stress distribution of the seven-hole titanium plate is located in the center of the center hole. During actual surgical procedures, clavicle fracture fragments and middle locking hole had stress superposition. If the titanium plate can avoid the stress concentration, it can effectively avoid the occurrence of the broken plate after implantation, provide theoretical guidance for clinical practice, and provide reference and technical route for biomechanical analysis of other types of titanium plate.

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

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