Objective:To investigate the roles of donor alveolar maerophages and the recipient circulating neutrophils in early-stage reperfusion injury of lung allograft,and to study the interaction between the 2 kinds of cells.Methods:Twenty pairs of size-and weight-matched adult mongrel dogs were randomly assigned to 4 groups:C(control),D(leukocyte-depleted blood reperfusion),M(maerophage inhibition)and DM(leukocyte-depleted plus macropbage inhibition).The 20 cases of left lung transplantations were performed by the same surgeon.All procedures were identical,except that the donors in Group M and DM received the macrophage inhibitor gadolinium chloride(14 mg/kg)intravenously 24 h before operation,and that the recipients in Group D and DM underwent initial 10 min reperfusion with leukocyte-depleted blood collected from donors'inferior vena cava. All lung allografts were reperfused for 2 h.Results:Compared with Group D and C,macrophage inhibition ameliorated PO_2/FiO_2 and mean pulmonary arterial pressure(mPAP)consistently after 30 min reperfusion in Group M and DM;the parameters of lung reperfusion injury(malonaldehyde activity,wet/dry ratio)at 120 min after reperfusion were also significantly improved(P

Objective To measure the learning passion of medical students and evaluating its in-fluencing factors. Methods Taking 879 medical students as research subjects to conduct a questionnaire survey according to specialty and grade stratified sampling. The questionnaire contained two parts, includ-ing learning passion scale and general situation questionnaire. The effective recovery rate was 69.28%, 609 valid questionnaires were recovered. SPSS 22.0 and AMOS 21.0 software were used for statistical analysis of data, analyzing the reliability and validity of the questionnaire with internal consistency reliability coeffi-cient and confirmatory factor analysis. The factors of learning passion of medical students were analyzed by logistic regression analysis. Results The questionnaire of learning passion of medical students contained 12 measurement items, including 2 dimensions: harmonious passion and obsessive passion. The questionnaire was of fine reliability (Cronbach's Alpha=0.916) and validity (χ2/df=3.401,RMSEA=0.073,GFI=0.958). The
learning passion of medical students was at upper middle level (4.390±0.934). The influence of achievement level (OR=1.691, 95%CI=1.415 to 2.021), school satisfaction (OR=0.586, 95%CI=0.402 to 0.854) and profession plan (short-term plan OR=2.121, 95%CI=1.310 to 3.434;long-term plan OR=3.822,95%CI=1.972 to 7.405) on learning passion were statistically significant. Conclusion The questionnaire of learning pas-sion of medical students has fine reliability and validity. Achievement level, school satisfaction and profes-sion plan are factors affecting the learning passion of medical students.

Objective To compare the acceptance of 2 different teaching modes－physicians and therapists teaching together and separately in continuing education students of rehabilitation medicine. Methods A questionnaire was filled by the students who attended the lectures of both Rehabilitation of Elbow Injury and Rehabilitation of Wrist Injury in the 9th National Orthopedic Class. The former lecture was taught by the rehabilitation physician and physical therapist together (together mode), and the latter lecture was taught by the rehabilitation physician and physical therapist separately (separate mode). The questionnaire included the choices and text questions. Results There were 45 copies of effective questionnaire all together. The satisfaction of both lectures were above 90%. As to teaching modes, 77.8% students liked together mode better, and 22.2% students preferd the separate mode. 93.9% students would or maybe use together mode in their future work, and 88.9% in separate mode. Conclusion The mode of physician and therapist giving lectures together is well accepted by students.

Enzyme-Linked Immunosorbent Assay ; Hepacivirus ; isolation & purification ; Hepatitis C ; virology ; Humans

OBJECTIVETo explore whether microRNA-200a (miR-200a) could be used as a novel biomarker of liver cancer using a rat model system.

METHODSDiethylnitrosamine abdominal injection was applied to induce liver cancer in the F344 rat strain (n =40); ten unmodeled rats served as controls. In addition, human subjects with normal healthy liver (n =10), liver cirrhosis (n =10), and liver cancer (n =10) were enrolled in the study. Blood samples from both rats and patients and rats' livers were collected for analysis. Real-time quantitative PCR and enzyme-linked immunosorbent assay were used respectively to measure the expressions of serum miR-200a and alpha-fetoprotein (AFP) for all rat and human subjects. In situ hybridization was used to detect the miR-200a expression in the rats' livers.

RESULTSComparison of normal rats and the liver cancer modeled rats showed that the latter had significantly lower expression of miR-200a (P less than 0.05), with decreasing expression following the progression of liver injury to cancer (liver cirrhosis rats less than early liver cancer rats less than advanced liver cancer rats); in contrast, the AFP levels were significantly higher in the liver cancer modeled rats only at the early and advanced stages of the liver cancer (P less than 0.05). These

RESULTSsuggested that miR-200a expression decreases during the developmental process of liver cancer, while AFP expression increases distinctly at the stage of tumor formation. Analysis of the human subjects' clinical samples showed that miR-200a expression was decreased in both liver cirrhosis patients and liver cancer patients (vs. normal liver subjects, P less than 0.05), while AFP showed abnormal expression only in the patients with liver cancer. Comparison of the normal rats and modeled rats using in situ hybridization showed the positive rates for miR-200a expression were 1.00% +/- 0.01% in rats with normal liver, 0.37% +/- 0.03% in rats with fibrotic liver, 0.14% +/- 0.01% in rats with cirrhotic liver, 0.05% +/- 0.00% in rats with early stage liver cancer, and 0.01% +/- 0.00% in rats with advanced stage liver cancer.

CONCLUSIONMiR-200a may play an important role in liver cancer development and may have diagnostic value for indicating early liver cancer.

Adult ; Aged ; Animals ; Case-Control Studies ; Female ; Humans ; Liver ; metabolism ; Liver Neoplasms ; blood ; metabolism ; Male ; MicroRNAs ; blood ; metabolism ; Middle Aged ; Rats ; Rats, Inbred F344 ; Young Adult ; alpha-Fetoproteins ; metabolism

OBJECTIVETo investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL).

METHODSThe human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology.

RESULTSThe flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles.

CONCLUSIONSThe proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.

Animals ; Cell Culture Techniques ; Cell Division ; Cells, Cultured ; Collagen ; Extracellular Matrix ; Fibroblasts ; cytology ; Humans ; Rats ; Skin ; cytology ; Tissue Engineering ; methods

OBJECTIVETo construct a multiple myeloma (MM)-specific APE1siRNA expression vector, and detect the specific knock-down effect of the siRNA on expression of APE1 protein.

METHODSAPE1siRNA cDNA sequence was designed, synthesized and inserted into pSilencer 2.0-U6 linear expression vector. pSilencer APE1siRNA was digested by enzyme EcoRI and BamHI, then linear vector and IgP fragments were conjugated by T4 DNA ligase. pSilencer IgP-APE1siRNA and pSilencer IE-IgP-APE1siRNA were digested by enzyme EcoRI or XhoI. Linear vector and IE or Kappa fragments were conjugated by T4 DNA ligase. Then a MM specific pSilencer K-IE-IgP-APE1siRNA was cloned. The recombinant products were identified by DNA sequencing and enzyme digestions at each step. pSilencer K-IE-IgP-APE1siRNA plasmid was transfected to KM3, HOS, MDA-231 cells by liposome. APE1 gene silence induced by RNAi was analysed by Western blot.

RESULTSAPE1 protein in KM3 cells could be knocked down effectively and specifically by pSilencer K-IE-IgP-APE1siRNA vector. After 2 days, the level of APE1 protein in KM3 cells transfected with siRNA was 0.118 +/- 0.047, while that transfected with plasmid only was 0.988 +/- 0.029. The efficiency of gene silence was 90%.

CONCLUSIONA MM specific APE1siRNA expression vector was successfully constructed.

Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Genetic Vectors ; genetics ; Humans ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

BACKGROUNDAn early identification of the composition of arterial thrombus may have diagnostic, therapeutic, and prognostic implications. The variation of magnetic resonance (MR) signal intensity between white and red thrombi, especially in the susceptibility sensitive MR sequence, remains unknown. Our research was to evaluate the feasibility of MRI in differentiating of white and red thrombi with a phantom study.

METHODSA total of 12 red and 12 white thrombi were prepared with the venous blood. Examination of the phantom was completed using a 3.0T MR unit, including fluid attenuated inversion recovery (FLAIR) T1, T2-weighted imaging (T2WI), FLAIR T2, T2 gradient echo (T2 GRE) imaging, and susceptibility weighted angiography sequences (SWAN). MR signal intensity patterns of the thrombi were objectively classified as hyperintensity, isointensity and hypointensity, compared with the background agar. The volume of thrombus was calculated and correlated with its signal intensity.

RESULTSFor white thrombi, 11/12 clots showed hyperintensity and 1/12 showed isointensity in FLAIR T1 images. In T2WI, 6/12 clots showed hyperintensity, 3/12 isointensity, and 3/12 hypointensity. In FLAIR T2, 8/12 clots showed hyperintensity and 4/12 showed isointensity. In T2 GRE, 3/12 clots showed hyperintensity and the remaining 9/12 clots showed isointensity. In SWAN, 5/12 clots demonstrated hyperintensity and 7/12 isointensity. For the red thrombus, 12/12 clots demonstrated hyperintensity in FLAIR T1, T2WI, and FLAIR T2 sequences. In T2 GRE and SWAN sequences, 3/12 clots displayed hypointensity and the remaining 9/12 clots showed slight hyperintensity. Thrombi with hypointensity displayed in T2 GRE and SWAN sequences were significantly larger than those with hyperintensity.

CONCLUSIONSDifferentiation of white and red thrombi with conventional MR sequence is unreliable, because both kinds of thrombi do not possess unique signal intensity features in these sequences. Red thrombus may or may not show hypointensity in the susceptibility sensitive MR sequences, depending on its size and time course.

Humans ; Magnetic Resonance Imaging ; methods ; Phantoms, Imaging ; Thrombosis ; diagnosis ; pathology

The study was purposed to explore the effect of panel reactive antibody (PRA) serum from patients with β-thalassemia on proliferation and apoptosis of the CD34(+)cells from cord blood and its mechanism. CD34(+) cells of umbilical cord blood were incubated with different sera and complement respectively. After incubation, the samples were centrifuged and the supernatants were collected for lactate dehydrogenase (LDH) detection, and the CD34(+) cells were harvested and measured for the apoptosis by flow cytometry with Annexin V/PI. The intracellular DNA synthesis were also quantified by [(3)H]TdR incorporation using liquid scintillation counter. The results showed that concentration of LDH in PRA positive groups was higher as compared with control group, and the DNA synthesis of CD34(+) cells in PRA positive groups were inhibited. There were no differences in the percentage of cell apoptosis and necrosis among different groups. It is concluded that thalassemic serum PRA impairs the cell membrane, inhibits the DNA synthesis, which can be increased by addition of the complement, but PRA had no significant effect on apoptosis of CD34(+) cells.
Antibodies ; blood ; immunology ; Antigens, CD34 ; Apoptosis ; immunology ; Cell Proliferation ; Child ; Fetal Blood ; cytology ; Humans ; beta-Thalassemia ; blood ; immunology ; pathology

Objective To explore the establishment of a rat model of acute radiation-induced liver injury and sig-nificance of the dynamic changes of TGF-β1 expression.Methods Forty healthy 6-week old male SD rats were randomly divided into model group (n=30) and control group (n=10).The right liver of rats in the model group was given a single dose of 25 Gy 6 MV X-ray irradiation.Histopathological examination using HE staining and transmission electron microsco-py were conducted to observe the liver pathological changes in rats at 3, 5, and 10 days after irradiation, serum TGF-β1 was detected, and relevant indicators of liver function ( ALT, AST, ALP) were determined.Statistical analysis was per-formed using SPSS 17.0 software.Results At 3, 5 and 10 days after irradiation, early pathological changes in the liver cells were observed by electron microscopy, the expression of TGF-β1 was gradually increased with the time prolongation, and significant differences were found between the model group and the control group at different time points (P<0.05). The light microscopic observation of liver tissues did not show significant differences between the control group and model group.The liver ALT, AST, ALP at different time points did not show significant differences between the two groups ( P>0.05).Conclusion Electron microscopy can be used to evaluate the early changes of radiation-induced liver injury, pri-or to the alterations visible by routine light microscopy.TGF-β1 can be used to predict the degree of radiation-induced liver injury, and may be used as a sensitive serum cytokine in predicting the degree of radiation-induced acute liver injury.