BACKGROUND: The precartilaginous stern cells are limited regarding in vitro proliferative capacity, but the immortalized cell lines can provide a large number of stable immortalized cells, and simian virus 40 large T antigen gene (SV40Tag) is one of gene fragments which are commonly used and effective in vitro immortalized ceils. OBJECTIVE: To construct human immortalized precartilaginous stem cells (IPSCs) using human precartilaginous stem calls induced by SV40LTAg gane. METHODS: The human immortalized precartilaginous stem calls were isolated from aborted fetus and purified with enzyme digestion and immunomagnetic beads screening method. By using liposome-mediated gene transfection technology, plasmid pCMVSV40T/PUR containing SV40Tag was transfected in primary embryonic precartilaginous stem cells, while non-transfected cells sewed as negative controls. Positive clones were cultured to observe the cell morphology and the passage recovery, to calculate cell survival rata and population doubling time, to drew call growth curve. Immunofluorescence cytochemistry was used to detect the expression of IPSCs fibroblast growth factor receptor 3, the expressions of SV40Tag and fibroblast growth factor receptor 3 in the human precartilaginous stem cells were determined by RT-PCR. RESULTS AND CONCLUSION: Morphology of human IPSCs seemed coincidence with primary human precartilaginous stem cells. The survival rate of human IPSCs was not influenced by subculture, freezing and recovery, but the survival rate was descended in the human precartilaginous stem cells at the 6~(th) and 10~(th) passages (P < 0.01). Compared with cells at the 6~(th) and 10~(th) passages, the proliferation of human IPSCs was greater, with short population doubling time and high growth rate (P < 0.01). The immunofluorescence showed that fibroblast growth factor receptor 3 was positive in human IPSCs at the second passage, and the RT-PCR results of fibroblast growth factor receptor 3 revealed a specific amplification band at 400 bp,.while that of SV40Tag revealed at 560 bp. No band was seen in the primary cells. It is indicated that SV40Tag human IPSCs can be constructed successfully using immunomagnatic bead screening technology and liposome transfection technique.

Animals ; Catechin ; analogs & derivatives ; pharmacology ; Disease Models, Animal ; Fatigue ; metabolism ; physiopathology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

OBJECTIVEOvertraining is a serious problem in sports, assessed by comprehensive multi-index evaluation, but so far there is still no sensitive, specific monitoring indicator or simple evaluation method to evaluate it. This research established a method for detecting plasma cell free DNA (cfDNA) of rats by real time PCR and discuss edits significance: a new molecular marker of overtraining?

METHODSTwelve male SD rats were randomly divided into control group and overtraining group. The overtraining group rats were undertaken overtraining on a motor-driven treadmill for 5 weeks, while the control group rats kept quiescent. All the rats were drawn blood at pre-and after-5 weeks to detect plasma levels of cfDNA, testosterone (T) and corticosterone (Cort) as well as peroxidation/antioxidation parameters (T-AOC, MDA, SOD, GSH-Px) and creatin kinase (CK).

RESULTS(1) Plasma cfDNA of rat was detected specifically by our real time PCR. (2) Compared with control group rats, the plasma cfDNA of overtraining rats increased obviously (about 5.43 fold). (3) Plasma cfDNA was related to plasma T, Cort, T/C ratio and MDA (correlation coefficent were -0.729, 0.854, -0.655 and 0.720, respectively) rather than plasma T-AOC, GSH-Px, SOD and CK.

CONCLUSION(1) A real time PCR method was established successfully to determine plasma cfDNA of rat. (2) A remarkable raises of plasma levels of cfDNA were found in overtraining rats which were associated with T, Cort and T/C, suggested that plasma cfDNA might be a new molecular marker of overtraining. (3) The increase of plasma cfDNA of overtraining rat might correlate with enhanced oxidative stress induced by overtraining instead of muscle damage.

Animals ; Biomarkers ; blood ; Corticosterone ; blood ; DNA ; isolation & purification ; Exercise Test ; Fatigue ; blood ; Male ; Physical Conditioning, Animal ; Plasma Cells ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Testosterone ; blood

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Female ; Herbicides ; poisoning ; toxicity ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Paraquat ; poisoning ; toxicity ; Risk Factors ; Time Factors ; Young Adult

Objective To detect the expression levels of vascular endothelial growth factor D ( VEGF-D) , D2-40 and MMP-7 in papillary thyroid carcinoma ( PTC) and to explore their relationship with micro lymphat-ic density, distribution,and cervical lymph node metastasis .Methods SP immunohistochemistry was used to study the expression of VEGF-D, D2-40 and MMP-7 in 38 cases of PTC with cervical lymph node metastasis , 45 cases of PTC without cervical lymph node metastasis , and 22 cases of normal thyroid tissues .Micro lymphatic density ( MLD) was counted .Results The expression level of VEGF-D in PTC with cervical lymph node metasta-sis and normal thyroid tissues had significant difference (χ2 =13.074, P=0.000, P<0.0167).The expression level of D2-40 in PTC with and without cervical lymph node metastasis had significant difference ( P<0.05 ) . MMP-7 expression in PTC with and without cervical lymph node metastasis had statistical significance ( 91.67%vs 73.33%, P<0.05).MLD in PTC with and without cervical lymph node metastasis and in normal thyroid tis-sues had statistical difference (11.7 ±3.5 vs 8.9 ±3.1 vs 3.9 ±2.7, P<0.05).Conclusion VEGF-D, D2-40 and MMP-7 are highly expressed in PTC and they are possibly related with cervical lymph node metastasis of PTC.

Objective To analyse the CT features of pulmonary echinococcosis and to assess CT value in diagnosing this disease.Methods CT fingings of pulmonary echinococcosis in 35 cases proved pathologically were analyzed retrospectively.Results Pulmonary granulosus echinococcosis was 33 cases including simple hydatid cyst (11 lesions),datugter cyst hydatid (3 lesions),with calcification (1 lesion) ,rupture (11 lesions),merging infection (4 lesions) and rupture into thorax (3 lesions).Pulmonary alveolar echinococcosis was 2 lesions,CT findings apperared as irregular,peripherally scattered more nodules with small vacuoles or calcification and in combination with hepatic alveolar echinococcosis.Conclusion Pulmonary echinococcosis had characteristic features on CT,CT examination can provide more imaging informations for the treatment of pulmonary echinococcosis

Objective To investigate the mechanism of ventricular dilation-induced arrhythmias by dilating isolated rat hearts. Methods Isolated rat hearts were perfused by Langerdorff method. After equilibration, 80 hearts were randomly divided into four groups as follows: (1) control group (n=20), (2) Ca2+ preconditioning (CPC) group (n=20), (3) streptomycin group (n=20), and (4) CPC + streptomycin group (n=20). A latex balloon which can be filled with fluid was anchored in the left ventricle through the left atrium and mitral valve. Epicardial ECG of the left ventricle, left ventricular pressure, coronary flow and heart rate were recorded before and during ventricular dilation by injecting fluid into the latex balloon. The rate and duration of ventricular dilation-induced arrhythmias were recorded. Results Under the same increase in ventricular end-diastolic pressure made by inflation of the balloon, the rate of arrhythmias was 100% and duration of arrhythmias was 2.56±0.46 s in the control group. Both the rates of premature ventricular beat (90 %) and ventricular tachycardia 70 % ) were high. Compared with the control group, the total rate (60 % ) of arrhythmias was lower, and duration (1.67±0.61 s ) of arrhythmias was shorter in the CPC group. Both the rates of premature ventricular beat (60%) and ventricular tachycardia (40%) were low comparatively. The rate of arrhythmias (45 %) was lower and duration ( 1.64±0.42 s)of arrhythmias was shorter, and the rates of premature ventricular beat (30 % ) or ventricular tachycardia (35 %) were lower in the streptomycin group than in the control one. The least ventricular dilation-induced arrhythmias occurred in the CPC + streptomycin group. The rate of arrhythmias (10%) was the lowest and duration (1.01±0.37s) of arrhythmias was the shortest; both the rates of premature ventricular beat (5%) and ventricular tachycardia (10%) were the lowest. Conclusions Ventricular dilation may induce arrhythmias in isolated rat hearts. Stretch-activated ion channel and the increase in [Ca2+]I are supposed to play important roles in the pathological mechanism.

Objective:To study the characteristics of high school students' coping styles when they faced frustrations and troubles,and the relationship between coping style and academic achievement.Methods:649 high school students completed questionnaire about middle school students' coping styles.Results:Girls were more inclined to use venting(2.7?0.7/2.4?0.7,F=23.32,P

Holmium ; Humans ; Kidney ; Lithotripsy, Laser ; Shock, Hemorrhagic ; Ureteroscopy

Chronic kidney disease (CKD) is a serious chronic disease with high incidence, poor prognosis, and a variety of complications. Indoxyl-sulfate (IS) and p-cresol sulfate (PCS) are two typical gut-derived uremic toxins, which are produced by the co-metabolism of intestinal microbes and the host. With the progression of CKD, gut-derived uremic toxins such as IS and PCS accumulate in patients with CKD and thereafter accelerate the progression of CKD. Gut microbiota is closely related with CKD, and targeting gut microbiota to regulate gut-derived uremic toxins synthesis and metabolic pathways may be a promising strategy to delay the progression of CKD. In this paper, the relationship between gut microbiota, gut-derived uremic toxins, and CKD was analyzed, and the strategy to delay the progression of CKD by targeting gut microbiota and uremic toxins metabolism pathway was proposed.