Humans ; Troponin I

Humans ; Myocardium ; chemistry ; Sensitivity and Specificity ; Troponin ; analysis

The authors described the procedures for the establishment of clone 515 of the cell strain of osteochondrosarcoma by capillary method.A single cell was sucked into a capillary tube by a microsyringe under the control of a micromanipulator.A segment of the capillary tube with the cell was then planted into the plasma clot in a Carrel flask. After the addition of conditioned medium,the flask was incubated at 37℃.Daily ob- servations were made.The propagation of the single cell,the migration of its progenies out of the capillary tube and the change in form and size of the cells were recorded. 63 days after the isolation,the progenies covered the whole bottom of the flask.First subculture was then made and the clone was established therefrom.

Objective To evaluate the growing condition of human umbilical vein endothelial cells (HUVECs), which were cultured on the membrane of different component, such as Chinese medicine, before or after the alkali treatment of 3-hydroxybutyrate-co-3-hydroxyhexanoate copolyesters (PHBHHx) and the biocompatibility between PHBHHx flim and endothelial cell. Methods The HUVECs were harvested by Baudine method,and identified by immunohistochemical method.Then the HUVECs of third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. After that, the morphology of HUVECs on different surfaces were observed by scanning electron microscopy, the distribution condition on different membranes were compared by cell-labeling immunofluorescence, and the cell viabilities of all groups were detected by MTT method. Results The HUVECs were successfully separated,and immunohistochemistry staining of FLK-1 and factor Ⅷ was positive.The result of HUVECs culture showed that cells on the material surface growed well, and proliferated significantly. The MTT analysis showed that the PHBHHx film of surface modification and adding some certain proportion of Chinese medicine could promote the growth and proliferation of HUVECs in vitro,and the cells were thriving, full shape, distribution on the surface by scanning electron microscopy and fluorescence microscopy.Conclusion The PHBHHx film of surface modification and containing certain proportion of Chinese medicine coating had good compatibility of HUVECs, which was favourable to cell growth, adherence and proliferation in vitro.

AIM To provide toxicokinetics data for toxicity studies of repeated doses of sodium 9-[2-(phosphonomethoxy) ethyl] adenine (PMEA-Na). METHODS The concentrations of PMEA-Na in plasma and urine were determined by HPLC/MS/MS method after single and multiple iv administrations in dogs. Data were executed by the statistical moment method to acquire the toxicokinetics parameters. Serum biochemical tests and histopathological examination were performed. RESULTS The system exposure of PMEA-Na in dogs was dose-dependent over the dose range of 1.0-6.0 mg·kg-1. The areas under the plasma concentration-time curve of PMEA-Na after single and multiple iv administrations at 1.0, 3.0 and 6.0 mg·kg-1 dosage were (2.3±0.5), (8.4±1.6), (17.5±3.7) and (5.0±0.4), (15.9±3.2), (30.3±4.7)mg·L-1·h, respectively. The urinary excretion of PMEA-Na in 72 h after iv administration was (87.0±4.8)% at the dose of 3.0 mg·kg-1. In 6.0 mg·kg-1 dose group, liver enzyme activity of glutamic-pyruvic transaminase and serum levels of total bilirubin, blood urea nitrogen, creatinine and triglycerides were all significantly elevated; glucose level significantly decreased comparing with the control group. Histopathological observation showed distinct pathological changes in liver and kidney tissues of 6.0 mg·kg-1 dose group. CONCLUSION There was evidence of toxicity after repeated-dose (14 d) of PMEA-Na in dogs and the major toxicity target organs were the kidney and liver.

Objective To assess the applicability of the Modification of Diet in Renal Disease (MDRD)formula to kidney function impaired Chinese diabetic patients.Methods Glomerular filtration rates(GFRs)in 463 Chinese diabetic patients(219 female,244 male,aged 14 to 88)were estimated by measuring ~(99m)Tc-DTPA clearance and with equations based on serum creatinine(Scr)and cystatin C(Cys C)concentrations.GFRs derived from various equations were compared with the ~(99m)Tc-DTPA clearance GFRs and their relative accuracies were assessed with ROC analysis.All the Scr measurements were performed with both the Roche enzymatic assay and the Beckman LX20 kinetic alkaline picrate assay,and Cys C with immunonephelometric and immunoturbidimetric assays.Results The reciprocals of Cys C and Scr were linearly correlated with ~(99m)Tc-DTPA clearance GFRs(r=0.830 and 0.690,repectively).The correlation of GFR with Scr could be expressed by an adjusted MDRD equation:GFR [ ml?(min?1.73 m~2)~(-1)]=175?(Scr)~(-1.154)?(age)~(-0.203)?0.742(female)?0.827,where 0.827 was a coefficient for Chinese.The adjusted equation showed a better accuracy than the MDRD equation(areas under the ROC curve 0.818 vs 0.644).The adjusted equation was also more accurate than equations obtained in previous Chinese studies.GFRs were also estimated by using Cys(in mg/L)with the following equation:GFR [ ml?(min?1.73 m~2)~(-1)] = 63.24?(Cys C)~(-0.3378).The accuracy of the Cys equation was similar to the Scr equation,or better in patients aged 60 and above.The Roche enzymatic results which were traceable to the isotope dilution mass spectrometry(IDMS)methods were significantly lower than Beckman LX20 results,but the results were closely correlated with each other(Y = 0.94X-0.02).When non-traceable Scr results were used,the coefficient needed to be adjusted.Conclusions GFRs can be estimated with equations based on either Scr or Cys C.GFR estimation should use standardized Scr results and take into account ethnic effects.

Recent clinical reports have demonstrated that the use of umbilical cord blood (UCB) opened a new source of stem cell for hematopoietic stem cell transplantation, leading to the development of cord blood banks world-wide. Prior to the large scale construction of UCB banks, quality control must be performed for health care providers and manufactures. With increasingly stringent regulatory requirement in blood industry, quality control is playing an important role in the operation of blood centers and stem cell laboratories. Reviewed the lectures in the biology of UCB and UCB banks published in recent years, our experiences were discussed in setting up Shandong blood bank to define process variables associated with the collection of UCB, to determine and optimize the procedures and materials used, to ascertain how UCB can be processed in clean room as mononucleated cell preparations, and to analyze using of long-term storage of UCB in research and clinic in the future. Our conclusions are: (1) the establishment of UCB banks for use in transplantation appears to be easy, effective and particularly suitable approach in China under cGMP conditions; (2) the procedures for volume reduction by closed and semi-automated blood processing system, SSP HLA typing, biocode and local computer net, microbiological tests and the 50 ml cryobags for storage constitute a cost efficient system for large-scale UCB banking; (3) the average of 60 ml UCB collection may contain sufficent marrow repopulating cells for children and most of adult recipients; and (4) hematopoietic stem and progenitor cells in cord blood have a more potent proliferative ability than those derived from bone marrow in cell expansion potentials.

ObjectiveTo research the relationship between the plasma levels of myeloperoxidase (MPO) and the onset and progress of acute coronary syndrome (ACS) and the severity of coronary lesions in patients with ACS.MethodsSeventy-eight patients hospitalized with chest pain were enrolled,including 41 patients with ACS (ACS group),17 patients with stable angina pectoris(SAP,SAP group) and 20 patients serving as control (control group).Forty-one patients undergoing coronary angiography were divided into single vessel lesions group (7 patients),double vessel lesions group (7 patients),multiple vessel lesions group ( 12 patients) and no vessel lesions group ( 15 patients) based on the vessel lesions of the left anterior descending,left circumflex artery and right coronary artery.According to the diameter stenosis of major coronary artery,there were 15 patients in no vascular stenosis group,2 patients in mild vascular stenosis group,6 patients in moderate vascular stenosis group and 18 patients in severe vascular stenosis group.The levels of MPO were measured by enzyme-linked immunosorbent assays(ELISA).ResultsThe levels of MPO in ACS group [( 252.10 ± 27.07 ) μ g/L]were higher than those in SAP group[( 185.81 ± 17.85 ) μ g/L]and control group [( 140.42 ± 71.40) μ g/L](P ＜ 0.05 ),the levels of MPO in SAP group were higher than those in control group(P＜ 0.05 ).The levels of MPO in single vessel lesions group and multiple vessel lesions group were higher than those in no vessel lesions group (P ＜ 0.05 ),but there was no significant difference among single vessel lesions group,double vessel lesions group and multiple vessel lesions group (P ＞ 0.05 ).The levels of MPO in mild vascular stenosis group,moderate vascular stenosis group and severe vascular stenosis group were higher than those in no vascular stenosis group (P ＜ 0.05),but there was no significant differenceamong mild vascular stenosis group,moderate vascular stenosis group and severe vascular stenosis group(P ＞ 0.05 ).A positive correlation was observed between the levels of MPO and neutrophils (r =0.288,P=0.018 ),creatine kinase isoenzyme-MB(r =0.469,P=0.043 ),subject groups( r =0.757,P=0.000),vessel lesions (r =0.584,P=0.000) and the degree of vascular stenosis (r =0.491,P=0.001).Conclusion MPO may predict ACS and reflect the severity of coronary lesions in ACS as a novel inflammatory marker.

The current study analyzed the data of 4 000 umbilical cord blood (UCB) units collected in Shandong Cord Blood Bank from the end of 1999 to March 2001. The averages of nucleated cells and CD34(+) cells were more than 1.2 x 10(9) and 3.9 x 10(6) per UCB unit respectively, and more than 1.5 x 10(9) nucleated cells per UCB unit were obtained in 768 UCB units. These UCB units are suitable for transplantation in patients with a body weight greater than 40 kg. The analysis of HLA gene frequency showed that A2, A24, A11, B13, B51, DR15, DR7 and DR9 are the common halotypes in Shandong population and similar to those in the other areas of China. 40% patients could search out at least 1 UCB unit with 1 mismatched HLA locus in Shandong Cord Blood Bank.
Antigens, CD34 ; immunology ; Blood Banks ; Blood Preservation ; Cell Count ; China ; Data Interpretation, Statistical ; Fetal Blood ; cytology ; immunology ; metabolism ; Gene Frequency ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; Humans ; Leukocyte Count ; Leukocytes ; cytology ; immunology ; Time Factors

The experience with the umbilical cord blood (UCB) stem cells for unrelated transplantation from our 3 000 UCB storage was described. UCB, collected from closed blood bags, were mixed with hydroxyethyl starch for nucleated cell (NC) enrichment. After finishing CD34 analysis, culture of hematopoietic progenitors (CFU-GM and CFU-GEMM) assays, microbial culture, HLA Class I (A, B) serology and class II (DR) low resolution SSP typing, cord blood units are stored in the liquid nitrogen for clinical applicatoin. Cord blood contained an average of nuclear cell (NC) (1.2 +/- 0.6) x 10(9), CD34(+) cells (3.0 +/- 3.7) x 10(6), CFU-GM (1.1 +/- 0.7) x 10(6) and CFU-GEMM (1.1 +/- 1.2) x 10(6) for storage and the recovery rates were 91%, 88%, 85% and 82%, respectively. The recovery rates for red blood cell and Hb were (39 +/- 9)% and (40 +/- 8)%, respectively. The storage volume was (35.1 +/- 7.1) ml in a 50 ml storage bags. The mean time from collection to processing of 15 hours (range 4 - 24 hours) had no influence on cell viability. The cell viability before processing is more than 95% and 92% after UCB thawing. The recovery rates of NC, CD34(+) cells and CFU-GM post-thawing were 96%, 90% and 91%, respectively. There were no HIV antibody (HIVAb) positive in all of UCB units. For an incidence of processed samples, infection with syphilis, HBsAg, HBcAb, HCVAb, CMV, bacterial contamination and abnormal hemoglobin were 0.1%, 0.8%, 3.2%, 0.2%, 87.1%, 1.2% and 0.1%, respectively. More than 3 HLA loci matched can be found for random patients in our cord blood bank and 6 HLA loci matched have 5%. For transplantation with nucleated cell counts of > 2.7 x 10(7) cells/kg, our cord blood bank will be able to provide all of the umbilical cord blood stem cell samples for children and 50% of units can be used for some of adult recipients transplantation in the country. It is concluded that: (1) The large cord blood banking for 20 000 UCB storage is feasible in China. (2) Our system of whole procedure and methods is functionable for supplying qualified cord blood units in transplantation. (3) The volume for collection is critical to the yield of CD34(+) cells or hematopoietic progenitor cells, however cord blood NC is also important and proportional with CD34(+) cells. Only the units containing more than 8 x 10(8) cells and more than 60 ml of cord blood can be in the procession for storage.