Capsid Proteins ; genetics ; therapeutic use ; Chicken anemia virus ; genetics ; Gene Transfer Techniques ; Genes, Viral ; Hep G2 Cells ; Humans ; Laryngeal Neoplasms ; genetics

In Chongqing Medical University,Introduction to Clinical Medicinewas first launched in 2002. In this article, based on the summary of 2014 to 2015, we summarized respectively from teaching material construction, evaluation system with combination of formative assessment and summative assess-ment, network support platform, teaching quality monitoring system, and student questionnaire survey and evaluation. Through the study of the course, 99.5% (836/840) of the students think their team conscious-ness and cooperation ability were improved, 94.2% (791/840) of the students consider this course can im-prove their innovative ability of self-learning, 87.0% (731/840) of the students think it is beneficial to the improvement of their self expression ability and more than 78.0% of the students think they have achieved the goal of early clinical contact, expanding knowledge, and enhancing the basic professional quality.

As urine flow dynamic test is widely used in clinical,merge detrusor dysfunction of benign prostatic hyperplasia treatment research also more and more.For normal detrusor contraction force and slight damage (Pdet ＞ 20 cmH2O) of the operative efficacy of BPH patients have reached a consensus of scholars both at home and abroad.For merger severely impaired detrusor contraction force (Pdet 0-20 cmH2O) at odds with the treatment of benign prostatic hyperplasia patients,in combination with related literature,now just urethral catheterization or colostomy,the transurethral resection of the prostate in the same period the bladder again after colostomy,the comprehensive treatment evaluation IⅡ transurethral resection of the prostate surgery and review these main treatment.

The objective of this study was to screen out the effective shRNA which can inhibit the gene expression of tumour necrosis factor-alpha (TNF-alpha), to construct the recombinant plasmid and to determine its sequence so as to provide the new approach for searching gene therapy of TNF-alpha related diseases. The primary macrophages were added into 15% DMEM, then cells were adjusted as 2 x 10(7) cells/L and were inoculated in 6-well plate with 3 ml/well, and were cultured at 37 degrees C in a fully humidified atmosphere with 5% CO(2). Cells were stimulated with lipopolysaccharide (LPS) and the concentration of TNF-alpha in the supernatant at different time points was determined by enzyme-linked immunosorbent assay (ELISA). The 5 synthesized DNA sequences which can be transcripted into shRNA were transfected into cells with lipofectamine 2000, then the cells were stimulated with LPS for 24 hours. The concentration of TNF-alpha in the supernatant and the expression of TNF-alpha mRNA were determined by ELISA and reverse transcription polymerase chain reaction (RT-PCR) respectively. The most effective shRNA was inserted into plasmid, and the recombinant plasmid was identified by sequence analysis. The results showed that the concentration of TNF-alpha in the supernatant reached peak after the stimulation with LPS for 24 hours. In the RNA interference group, the shRNA 1 was the most effective one, which could inhibit the expression of TNF-alpha by 59.46% and the expression of TNF-alpha mRNA by 61.2%. The recombinant plasmid was cloned and the sequence of interest was obtained. In conclusion, the most effective shRNA targeting TNF-alpha was successfully screened out and the recombinant plasmid was constructed. The recombinant plasmid may be helpful to search new gene therapy for TNF-alpha related disease.
Animals ; Cells, Cultured ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Macrophages ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Recombination, Genetic ; Transfection ; Tumor Necrosis Factor-alpha ; genetics

The aim of this study was to explore the effect of hyperbaric oxygen (HBO) preconditioning on the rejection of skin allograft in mice and its molecular mechanism. BALB/c donor mice and C57BL/6 recipients received hyperbaric oxygen preconditioning once a day for 7 days. After skin transplantation, the recipients were treated with cyclosporine A (CsA) intraperitoneally. Immunofluorescent staining technique and flow cytometry were used to observe the influence HBO on percentage of spleen lymphocytes CD3+, CD4+, CD8+ and cell adhesion molecule LFA-1 (CD11a/CD18). The results showed that as compared with control, the numbers of CD3+, CD4+, CD8+, CD4+CD11a+, CD4+ CD18+, CD8+CD11a+, CD8+CD18+ lymphocytes of spleen decreased in HBO preconditioning groups and CsA group, and decreased markedly in HBO preconditioning combined with CsA group (p<0.05); the general state of recipient mice in HBO preconditioning combined with CsA group was better than that of recipient mice received HBO preconditioning or CsA only. It is concluded that the method of HBO preconditioning combined with traditional immunosuppressive agent CsA has remarkable advantage in inhibiting the rejection of skin graft. Its molecular protective mechanism is correlated with the expression of adhesive molecules on T cell subsets.
Animals ; Cell Adhesion ; Cell Adhesion Molecules ; pharmacology ; Cyclosporine ; pharmacology ; Female ; Graft Rejection ; prevention & control ; Graft Survival ; Hyperbaric Oxygenation ; Lymphocyte Count ; Lymphocytes ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Skin Transplantation ; immunology ; Spleen ; cytology ; Transplantation Conditioning ; methods

The objective of this study was to investigate the function and mechanism of hyperbaric oxygen (HBO) in antagonizing acute graft-versus-host disease (aGVHD) and improving the rate of survival. The lethally irradiated C57BL/6 recipients were injected with bone marrow and lymphocyte of spleen from BALB/c donors and were treated with HBO, cyclosporine A (CsA) and methotrexate (MTX). T lymphocytes and subsets, adhesion molecules and cytokines were detected by flow cytometry, ELISA and RT-PCR respectively. The results showed that the survival rate in HBO group was much higher than that in allogenetic bone marrow transplantation (allo-BMT) group and CsA + MTX group; the numbers of CD3(+), CD4(+), CD8(+), CD4(+)CD11a(+), CD4(+)CD18(+), CD8(+)CD11a(+), CD8(+)CD18(+) lymphocytes in spleen were decreased markedly by HBO and CsA + MTX (p < 0.05); the levels of IL-2 and TNFalpha mRNA and their serum concentrations in HBO group were much lower than those in allo-BMT group but were higher than those in CsA + MTX group; the levels of IL-4 and IL-10 mRNA in HBO group were much higher than those in allo-BMT group and CsA + MTX group. It is concluded that HBO has more remarkable advantage in improving the rate of survival than CsA + MTX, its mechanism of anti-aGVHD is tightly correlated with the transform of T cell and its subsets and the expression of adhesion molecules and cytokines.
Acute Disease ; Animals ; Bone Marrow Transplantation ; adverse effects ; Cytokines ; biosynthesis ; Female ; Graft vs Host Disease ; etiology ; therapy ; Hyperbaric Oxygenation ; Lymphocyte Transfusion ; adverse effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes ; immunology ; Whole-Body Irradiation

One patient confirmed as pituitary carcinoma by histopathology was retrospectively analyzed according to the great changes of tumor invasive growth in imaging examine and clinical characters. Clinically,pituitary carcinoma quickly grows,and the growth character of tumor is helpful in the diagnosis. Moreover,the treatment for pituitary carcinoma is different from pituitary adenoma.The rapid progress of pituitary adenoma indicates the possibility of primary carcinoma.More experience should be accumulated for the diagnosis and treatment of the disease.

Objective To study the expression of mRNA of OX40 and OX40L in the sciatic nerve,spleen,peripheral blood mononuclear cells and lymph nodes of EAN under the influence of Rho-kinase inhibitor.Methods All 54 female Lewis rats were divided into 3 groups:the EAN group,the EAN+ Rho-kinase inhibitor group and the complete Freund's adjuvant(CFA)group.The rats were sacrificed at 9,17 and 26 days after immunized.Ox40 and OX40L mRNA were detected by RT-PCR which came from spleens,sciatic nerves,peripheral blood mononuclear cells and lymphonodes.Results In EAN+ Rho-kinase inhibitor group,the mRNA expression of OX40 were 0.266±0.031,0.298±0.024 and 0.113±0.018 at 9.17 and 26 days in the sciatic nerve,the expression were 0.453±0.030,0.496±0.100 and 0.220±0.016 in the lymph nodes.The mRNA expression of OX40L were 0.247±0.018.0.298±0.026 and 0.165±0.013 in the sciatic nerve,the expression were 0.283±0.027,0.306±0.011 and 0.161±0.012 in the lymph nodes.The mRNA expression of OX40 and OX40L in EAN+Rho-kinase inhibitor group was lower than EAN group at the three time points(t=2.24-4.89,P＜0.05),and the demyelination and inflammation cells infiltrating were ameliorated in spinal nerve.CFA group didn't show any clinical manifestation.Conclusion Rho-kinase inhibitor may ameliorate tlIe development of EAN through inhabiting the OX40 and OX40L activation.

We retrospectively analyzed clinical and imaging data of 26 children with split cord malformations (SCMs). Based on Pang's classification, 14 SCMs were defined as type Ⅰ and 12 as type Ⅱ.Neural function was markedly improved in 20 patients postoperatively. Three of 4 children who did not undergo surgical treatment had neural function deteriorated. Two children lost follow-up. We suggest that Pang's Classification of SCMs may be useful in describing pathological changes and guiding surgical procedure; imaging examine (including MRI, CT and X-ray) would play a significant role in confirmed SCMs diagnosis; and surgical operation should focus on eliminate and prevent spinal cord damnification.

Objective To study the effect of Rho-kinase inhibitor on experimental allergic neuritis. Methods 54 female Lewis rats were divided into three groups; EAN group, EAN + Rho-kinase inhibitor group, and CFA group. The rats were sacrificed on the 9th day, 17th day, and 26th day after immunized. The changes of weight, EAN incidence, and mean day of onset, mean maximum clinical score, and histopa-thology were observed. Results The clinical course in EAN group reached peak on the 17th day. Compared with EAN group, the weights of Rho - kinase inhibitor group were increased, while EAN incidence, mean day of onset delay, and the clinical scores in Rho-kinase inhibitor group were significantly decreased, ( P < 0.01) , and the demyelization and inflammation cells infiltrating was ameliorated in spinal nerve. CFA group didnt show any clinical manifestation. Conclusions Rho - kinase inhibitor may ameliorate the development of EAN through inhabiting the Rho/ROK signal pathway.