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Objective T o investigate the tim e-dependent expression of m etallothionein (M T ) 1A m R N A and M T 2A m R N A in contused skeletal m uscle of rats. Methods A total of 54 Sprague-D aw ley rats w ere used in this study. T he rats w ere divided into tw o parts:control group (n=6) and contusion groups (0.5, 1, 6, 12, 18, 24, 30, and 36 h after contusion, n=6). T otal R N A w as extracted from skeletal m uscle. T he expression levels of M T 1A m R N A and M T 2A m R N A w ere detected by SY B R G reen I real-tim e PC R . Results T he expression trends of the tw o potential m arker genes w ere related to w ound age. In ad-dition to 0.5 h, there w ere significant contrasts betw een the control group and contused group (P<0.05), about the expression levels of M T 1A m R N A and M T 2A m R N A in different phases. A s the extension of w ound age, the relative expression of M T 1A m R N A and M T 2A m R N A at 1 h, 6 h, 12 h and 18 h after contusion dem onstrated upgrade tendency until its expression levels in 18 h peak w ith 239.41±15.20 and 717.42 ±50.76, respectively. W hen tim e extends to 24 h after injury, the expression of above tw o m arks decreased, respectively. T he M T 1A m R N A and M T 2A m R N A expression levels increased at 30 h and then decreased. Conclusion D eterm ination of M T 1A m R N A and M T 2A m R N A levels by real-tim e PC R m ay be useful for the estim ation of w ound age.
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Objective To study the relationship between psychological capital and quality of life (QL) in cancer patients and search for better corresponding nursing strategies. Methods Investigating psychological capital state in cancer patients, and analyzing the relationship between social support score and QL. Results Cancer patients′ positive psychological capital score were low: The total score were 74.33 ± 20.04, self-efficacy 20.87 ± 6.64, hope 22.12 ± 6.04, toughness 20.35 ± 5.65, and optimism 12.36 ± 5.55. Cancer patients had low quality of life, quality of life scores were 59.57±0.57, body condition 15.32± 5.90, psychological condition 47.74 ± 10.22, social function 20.46 ± 5.19,family situation 18.93 ± 8.89, total score 115.48±20.19;The psychological capital and quality of life of cancer patients were positively related (r = 0.517, P < 0.01), cancer patients dimensions of psychological capital was positively correlated with the dimensions of quality of life (r =0.189-0.517, P < 0.01). Conclusions Cancer patients′ positive psychological capital and the quality of life both need to be improved; Cancer patients′ psychological capital are closely related to the quality of life and we should pay attention to improve the patients′psychological capital so as to better their quality of life during the care process.
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Objective Take frizzled class receptor 4(FZD4) as an example for exploring whether the transmembrane receptor protein is a suitable marker for wound age estimation. Methods A total of 78 Sprague-Dawley rats were divided randomly into the control group and the contusion groups of 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 36h, 40h, 44h, and 48h after injury (n=6). Using a drop-ball technique, a contusion was produced in the right lamb of rats. The samples were dissected from injury zones. Then the expression of FZD4 was detected using immunofluorescence, real-time PCR and western blot, respectively. Results FZD4 was located on the membrane of skeletal muscle cells. Compared to the control group, FZD4 mRNA showed a signiifcant increase (over 2-fold) in 8h, 12h, 36h, and 40h after injury by Real-time PCR (P<0.05), and FZD4 protein showed a statistical up-regulation (less than 2-fold) in 8h, 36h, 40h, 44h, and 48h post contusion by western blotting (P<0.05). Conclusion The expression of FZD4 mRNA and protein are both time-dependent during contused skeletal muscle repair. To some degree, FZD4 mRNA was more suitable than corresponding protein for determining wound age.
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Objective T o investigate the relationship betw een the expression of secreted frizzled-related protein 5 (SFR P5) m RNA and the tim e interval after skeletal m uscle injury in rats by real-tim e PC R . Methods A total of ninety SD rats w ere random ly divided into the contusion groups at different tim es including 4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h after contusion, incision groups at different tim es including 4h and 8h after incision and the control group. T he sam ples w ere taken from the contused zone at different tim e points. T he total RNA w as isolated from the sam ples and re-versely transcribed to analyze the expression levels of SFRP5 m RNA . Results C om pared to the control group, the expression of SFRP5 m RNA in contusion groups w ere dow n-regulated w ithin 48 h after con-tusion and reached the low est level at 20 h, and the expression of SFRP5 m RNA gradually increased from 20 h to 48 h after contusion. T he expression of SFRP5 m RNA in the incised groups w ere signifi-cantly low er than that of the contusion groups at 4 h after injury. A t the tim e of 8 h, the expression levels betw een the contusion and incision groups show ed no statistically significant difference. Conclusion It is suggested that SFRP5 m RNA analysis m ay show regular expression and can be a m arker for estim ation of skeletal m uscle injury age.
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Objective In order to understand which kind of function genes play an important role for es-timating wound age, the variation of difference genes’ mRNA expression were compared after injury. Methods T he mRNA expression levels of seven candidate genes (ICAM-1, NF-κB, MX2, MT1, MT2, sTnI, and Cox6c) were analyzed in contused rat skeletal muscle at different time points using real-time fluorescent quantitative PC R (R T-qPC R ). T he rawC t values were normalized relative to that of RPL32 mRNA , and converted to standard C t values. A t each time point after injury, the standard deviations (SD ) of the standard C t values were calculated by SPSS. Results T he expression trends of the seven genes were all found to be related to wound age, but there were lower variation coefficients and greater reliability of sTnI and Cox6c when compared with other genes. Conclusion T he genes encoding struc-tural proteins or proteins that performbasic functions can be suitable for wound age estimation.
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Objective T o investigate the relation betw een injury tim e and the expression of cytochrom e c oxidase subunitⅥc (COX6C) m R N A in skeletal m uscle of rat after contusion. Methods A total of fifty-four SD rats w ere divided into the control group and the contusion groups (0.5,1,6,12,18, 24, 30, and 36 h after contusion), random ly. T he contusion m odel w as established by free fall drop of gravity ham-m er. A t corresponding tim e point after contusion, the regular histology w as exam ined and expression level of COX6C m R N A w as tested by real-tim e PC R after extraction of total R N A from the tissues. Results T he m ain pathological features of 6 h after injury included edem a and hem orrhage in m yocytes w ith no inflam m atory cells found. A fter 6 hours, the findings included m yocyte degeneration and necro-sis, inflam m atory cells infiltration, and fibrous connective tissue proliferation in the contused zone. T he expression level of COX6C m R N A w as higher than that of the control group w ithin 6 h after contusion. T he expression level w as low er than that of the control group from 6-36 h after contusion. Conclusion T he level of COX6C m R N A expresses in a regular w ay after contusion. It m ay be useful for estim ating w ound age in com bination w ith the results of pathological features.
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Unleashed growth of gross healthcare expenses has emerged a major headache for public administration of countries at large.In China as well,such a rapid growth is incurred mostly by overtreatment due to irrational healthcare compensation.Against this background,it is highly significant to have a brand new insight into over-treatment in China' s viewpoint,and to further study the role of an improved mechanism for primary healthcare compensation for the purpose of curbing over-treatment.These efforts will help improve the primary healthcare service and realize goals of the ongoing health reform.
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<p><b>OBJECTIVE</b>To seek an appropriate culture condition in which porcine hepatocytes can continuously proliferate and express their normal differentiation phenotypes for a longer period of time.</p><p><b>METHODS</b>Different types and dosages of reagents, transferrin, epidermal growth factor and nicotinamide were used to culture porcine hepatocytes in serum-free Dulbecco's modified Eagle's medium (DMEM). And the proliferative effects and functions of the cells were detected at different culture times.</p><p><b>RESULTS</b>Transferrin at 5 ng/ml, nicotinamide at 10 mmol/L and epidermal growth factor at 10 ng/ml had better effects on the viability of hepatocytes than DMEM, DMEM + 10% fetal cattle serum FCS and other dosages of these reagents (P < 0.05). OD values of MTS were still high in culture at day 7 and day 10, while nearly 30% - 35% cells went into the S phase. Good hepatocyte functions were found in these groups, and the secretion of albumin was positively correlated with OD value of MTS. The levels of aspartate transaminase and ammonia in these media were lower than those in DMEM and DMEM + FCS.</p><p><b>CONCLUSION</b>Transferrin at 5 ng/ml, epidermal growth factor at 10 ng/ml and nicotinamide at 10 mmol/L are beneficial to the viability and proliferation of hepatocytes.</p>
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Animals , Cell Division , Cell Survival , Cells, Cultured , Epidermal Growth Factor , Pharmacology , Hepatocytes , Cell Biology , Niacinamide , Pharmacology , Swine , Transferrin , PharmacologyABSTRACT
Objective To study the effects of apoptosis associated factors Fas/Fas, Bcl-2/Bax, adhesion molecules including ICAM-1, VCAM-1, PECAM-1, P-selectin, L-selectin and E-selectin in rat heart allograft immune response at different time points. Methods Rat acute cardiac allograft rejection model (PVG heart to DA rats) were established. Heart samples were collected on the day 1, 3, 5, 7 postoperatively ( n =3 in each group). Apoptosis was monitored by TUNEL. Fas, FasL, ICAM-1 and Selectin was examined by means of immunohistochemistry. The expression of Bcl/Bax mRNA, VCAM-1 mRNA, PECAM-1 mRNA was detected by means of situ-hybridization. Results In acute cardiac allograft rejection group, the expression of apoptosis, Fas/FasL, Bax, ICAM-1, VCAM-1 and PECAM-1 was increased. Apoptosis, Fas/FasL and Bax were mostly detected in cardiomycotes and ICAM-1, VCAM-1 and PECAM-1 were mostly detected in vascular endothelium and infiltrated cells. There was no expression of apoptosis associated factors and adhesion molecules in normal rat heart.Conclusion The high expression of apoptosis, Fas/FasL, Bax, ICAM-1,VCAM-1, PECAM-1 may be related with cardiac allograft rejection.
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We investigated the mechanism of tolerance induction by identifying differences in infiltrating cell population and Ig subclasses deposition between tolerant and rejecting rat liver allografts.There were occasional minor differences in the T cell or CD11 b/c+(monoeyte/macrophage/granulocyte)infiltrate.By contrast,there was a major difference in B cell infiltrate with more B cells in tolerant grafts.IgM was increased in both tolerant and rejecting livers while deposition of IgG1 was observed in rejecting but not in tolerant livers.
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Objective To investigate the effects of apoptosis, Fas/FasL and Bax/Bcl-2 in rat liver allograft immune response. MethodsDifferent rat allograft models were established:(1)Acute liver allograft rejection group: PVG liver to Lewis rats; (2) Liver allograft tolerance group: PVG liver→DA rats; (3) F1(P/D)→D group: F1(PVG-DA) liver to DA rats; (4)HLTx(P/2→D)group: PVG/2 liver→DA rats. Liver samples were collected on days 1, 3, 5, 7 postoperatively (each n=3). Apoptosis was monitored by TUNEL(terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick-end-labeling). The expressions of Fas/FasL and Bcl-2/Bax mRNA were examined by means of immunohistochemistry and in situ-hybridization respectively. Results Compared with liver allograft tolerance group, in acute liver allograft rejection group, the apoptosis cell numbers increased in hepatocytes, FasL expression was high in infiltrated cells in portal tract areas and low in hepatocytes, and the expression of Fas and Bax mRNA increased. The expression of molecules detected in F1(P/D)→D group was similar with those in acute liver allograft rejection. The expression of molecules detected in HLTx(P/2→D)group was similar with those in liver allograft tolerance group. On the fifth day postoperatively, the apoptosis index in portal tract areas had a positive correlation with the expression of FasL in hepatocyte and a negative correlation with the expression of Fas and Bax. Conclusions The high expression of FasL in infiltrated cells of portal tract areas, Fas and Bax in hepatocytes may be involved in rejection response. The high expression of apoptosis in infiltrated cells of portal areas, FasL in hepatocytes may relate with the establishment of tolerance.
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Objective To explore the kinds and levels of phthalates leaching from disposable plastic products. Methods Samples of peritoneal dialysis solution, blood preservative solution, infusion instruments, preservative film, disposable plastic bags and water in plastic bottles were analyzed for phthalates by RP-HPLC after liquid-liquid extraction and/ or solid phase extraction. Results Di(2-ethylhexyl) phthalate (DEHP) was leached from all medical instruments, the maximum level of which reached 77.51?g/ L. Di-n-butyl phthalate was leached from disposable plastic bags, the level of which reached 91.45?g/ kg. Phthalates were not found in samples of preservative film and water in plastic bottles. Conclusion As DEHP leaching from the medical instruments might directly enter the human body, attention should be paid to its health hazards.
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Objective To study the toxicokinetics of phthalates in male rabbits. Methods Concentrations of DEHP or DBP in plasma of 6 healthy male rabbits were determined by RP-HPLC after constant rate infusion of 0.5 g/kg DEHP or DBP(IV). The toxicokinetic parameters were computed by 3P87 program. Results The toxicokinetic model of DEHP and DBP were both first-order elimination and two-compartment model with constant rate infusion. The main toxicokinetic parameters of DEHP were as follows: distribution phose t1/2(?)=0.101 h; elimination phase t1/2(?)=12.701 h; CLs=0.013 g?kg-1?h-1. The main toxicokinetic parameters of DBP were as follows: t1/2(?)=0.441 h; t1/2(?)=31.311 h; CLs=0.021 g?kg-1?h-1. Conclusion DEHP and DBP were both first order elmination and two-compartment medol with constand rate intusion(IV). DEHP and DBP could be rapidly eliminated in male rabbits.
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Objective To investigate a method for postmortem diagnosis of the sudden death from hypersensitivity by detection of serum tryptase and IgE.Methods Venous blood samples were collected from 9 cases of sudden death from hypersensitivity,19 cases of death from other causes and 10 health living adult individuals.The serum levels of the mast cell tryptase(MCT)and IgE were detected with immunofluorescent enzyme assay and enzyme linked immunosorbent assay(ELISA)respectively.MCT in the lungs was detected by immunohistochemistical method.Results The serum level of both MCT and IgE in cases of sudden death from hypersensitivity were higher than those in health living adults and cases of death from other causes(P﹤0.01).However,there was significant difference(P﹥0.05)in the serum level of IgE and MCT between the health living individuals and cases of death from other cause.More cells containing MCT-positive granules were detected in the lung in cases of sudden death from hypersensitivity than in cases of death from other causes(P
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Objective To investigate the application of orthogonal design to optimize real-time quantitative PCR conditions and perform it for ICAM-1.Methods After model of a skin injury was established in rats,total RNA was isolated from the injured skin tissue and reverse transcriptase reactions were carried out.Using orthogonal design real-time quantitative PCR conditions,the expression level of ICAM-1 gene was studied by use of cDNA as template.The standard of score was set up to amplification and dissociation curves and statistical analysis was done based on the optimized conditions.Results Through 25 tests the best real-time quantitative PCR conditions for ICAM-1 are anneal temperature at 61 ℃,0.4?g of cDNA and concentration of 150?mol/L each primer in the final reaction.There are no interaction among above factors.Conclusion Orthogonal design is a feasible,quick and economical method to optimize real-time quantitative PCR conditions.
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To investigate and analyze the factors which influence the public attitudes towards autopsy after accidental deaths.The study has carried on an investigation among 386 individuals in random,with a questionnaire named "The cognition of the public whether to carry on the autopsy after accidental deaths".Using the statistical methods of the Logistic Regression analysis and optimum regression analysis to analyze the recycling questionnaires.Through the diagnosis,we found there are four main factors which influence public opinion upon the autopsy after accidental deaths,the knowledge of autopsy,the believing in autopsy,years of schooling and family financial circumstances.