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 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

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

OBJECTIVETo study clinical effects of the over-articular external fixator combined with limited internal fixation for the treatment of Pilon fractures caused by high energy.

METHODSFrom September 2003 to April 2011, 36 patients with Pilon fractures caused by high energy were treated with the over-articular external fixator combined with limited internal fixator. There were 25 males and 11 females, ranging in age from 16 to 72 years old,with an average of 38 years old. The diagnoses of all patients were determined by conventional X-ray examination or three-dimensional spiral CT examination. The AOFAS scoring criteria was used to evaluate the therapeutic effects. The patients with comminuted fractures were treated with screw or Kirschner wire fixation without uncovering periost so as to enhance stability between fracture end and bone blocks,followed by the fixation with over-articular external fixators.

RESULTSAll the patients were followed up, and the duration ranged from 4 to 27 months, with an average of 13 months. Thirty-two patients got wound healing at the first stage. And the bone union duration ranged from 2 to 6 months, with a mean of 3 months. According to the AOFAS ankle-hindfoot subjective scoring standard, 13 patients got an excellent result, 20 good and 3 fair, with an score of 88.2 +/- 3.6. Twelve patients had infections at pinhole, 5 patients had pinhole pain. One patient had the fixator broken induced by over loading, who was cured after treatment. There were no complications such as nerve or vascular injuries, or osteomyelitis.

CONCLUSIONThe over-articular external fixation combined with limited internal fixation for the treatment of Pilon fractures caused by high energy is an ideal method, which has such advantages as reliable fixation, simple operation, coincidence with principles of biomechanical fixation, and benefit for fracture healing.

Adolescent ; Adult ; Aged ; Ankle Injuries ; diagnostic imaging ; surgery ; Ankle Joint ; diagnostic imaging ; surgery ; External Fixators ; Female ; Fracture Fixation ; Fracture Fixation, Internal ; Humans ; Internal Fixators ; Male ; Middle Aged ; Radiography ; Treatment Outcome ; Young Adult

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 investigate the diagnostic value of twinkling artifact in urinary calcium stones.Methods Calcium oxalate monohydrate,hydroxyapatite and whitlockite stone phantoms were prepared and embedded in porcine kidneys for ultrasound scanning.The length and width of twinkling artifact were measured.The intensity of twinkling artifact and the contrast-to-noise ratio (C/N ratio) of acoustic shadowing were recorded and all the data was analyzed statistically.Results All the stone phantoms generated twinkling artifact and acoustic shadowing.The difference of the intensity,length and width of twinkling artifact and the C/N ratio of acoustic shadowing had statistically significant each composition (P ＜0.01).There was no overlap between 95％ confidence interval of the intensity of twinkling artifact for any two compositions,and any two compositions of stones could be differentiated by the intensity of twinkling artifact (P ＜0.05).The C/N ratio of acoustic shadowing of calcium oxalate monohydrate and whitlockite stone phantoms were higher than those of hydroxyapatite ones (P ＜0.05).The receiver operating characteristic (ROC) curve displayed that the cutoff value of 1 127 of the intensity of twinkling artifact could be used to diagnose hydroxyapatite stones.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and area under the curve (AUC) were 74 ％,68 ％,69.8％,72.3％ and 0.743 respectively.Conclusions Twinkling artifact could be used to differentiate the calcium stones,which will be helpful for clinical treatment.

Objective To quantitatively analyze the value of color Doppler twinkling artifact and CT in distinguishing urinary stone composition of uric acid from that of cystine,and to assess the ability of color Doppler twinkling artifact and CT to diagnose urinary stone composition of uric acid.Methods Artificial stones made of uric acid and cystihe were inserted in porcine kidneys respectively.Color Doppler ultrasound and CT scanning were performed in vitro and twinkling artifact videos were recorded.The CT attenuation value,the width and length of twinkling artifact (TAW,TAL) were measured.The color pixels representing twinkling artifact intensity (TAI) were calculated.The differences between artificial cystine and artificial uric acid stones in TAW,TAL,TAI and CT attenuation value were compared.The receiver operating characteristic (ROC) curve was used to assess the ability of TAW,TAL,TAI and CT attenuation value to diagnose urinary stone composition of uric acid.Results The TAW [(4.16 ± 1.54)mm vs (4.97 ± 1.93)mm],TAL[(3.75 ± 1.05)mm vs (5.12 ± 2.51)mm] and TAI [(3 005.50 ± 812.33) vs (5 433.86±2 505.81)] of artificial cystine stones were less than those of artificial uric acid stones while the CT attenuation value of artificial cystine stones [(573.75 ± 110.10)HU vs (364.09 ± 67.28)HU] were greater (P ＜0.05).The ROC curve was displayed to diagnose urinary stone composition of uric acid,and the area under curve of CT attenuation value was bigger than that of TAI (0.936 vs 0.817,Z =2.308,P =0.021).Combination of color Doppler twinkling artifact and CT value would dramatically increase the diagnostic accuracy to 98％.Conclusions Both color Doppler ultrasound and CT were reliable examinations for distinguishing urinary stone composition of cystine from that of uric acid.Combination of color Doppler ultrasound and CT would be good references to diagnose urinary stone composition of uric acid.