OBJECTIVESTo study on the changes of intracellular calcium and magnesium in cirrhosis and its clinical significance.

METHODSThe calcium and magnesium were determined in serum (SCa, SMg), platelets (PCa, PMg), mononuclear cells (MNCCa, MNCMg), polymorphonuclear cells (PMNCa, PMNMg) and erythrocytes (RCa, RMg) of 50 patients with uncompensative cirrhosis (group A) and of 35 patients with compensative cirrhosis (group B). 35 health persons were the control group.

RESULTSThe SCa and SMg of group A were lower significantly than those of both group B and control group. The MNCCa, PMNCa, RCa, PMg, MNCMg, PMNMg, RMg of group A [(4.76+/-1.91) micromol/10(9), (7.56+/-2.88) micromol/10(9), (0.66+/-0.13) mmol/L, (5.53+/-2.25) micromol/10(11), (6.64+/-3.53) micromol/10(9), (10.12+/-4.32) micromol/10(9), (2.02+/-0.76) mmol/L] and those of group B [(5.34+/-2.41) micromol/10(9), (8.32+/-2.34) micromol/10(9), (0.67+/-0.11) mmol/L, (5.55+/-2.67) micromol/10(11), (6.56+/-3.44) micromol/10(9), (10.95+/-4.45) micromol/10(9), (2.21+/-0.74) mmol/L] were lower significantly than those of control group [(6.86+/-2.02) micromol/10(9), (9.89+/-3.23) micromol/10(9), (0.72+/-0.10) mmol/L, (7.43+/-2.78) micromol/10(11), (8.68+/-4.1) micromol/10(9), (13.96+/-5.76) micromol/10(9), (2.74+/-0.92) mmol/L]; t (group A vs. control group)=4.88, 3.48, 2.31, 3.45, 2.46, 3.52, 4.00, 0.01, 0.01, 0.05, 0.01, 0.02, 0.01, 0.01; t (group B vs. control group)=2.87, 2.34, 2.00, 2.89, 2.33, 2.45, 2.65, 0.01, 0.05, 0.05, 0.01, 0.05, 0.02, 0.02. The PCa of the patients with hepatic encephalopathy was higher, the SMg, PMg, MNCMg, PMNMg and RMg were lower than those of the patients without hepatic encephalopathy significantly. The SCa, SMg, PMg, MNCMg, PMNMg and RMg of the patients in Child stage C were lower significantly than those of the patients in Child stage B. There were no significant differences of PCa, MNCCa, PMNCa and RCa between Child stage C and Child stage B. There were no significant differences of SCa, MNCCa, PMNCa and RCa between the patients with and without hepatic encephalopathy. The ratios of PCa/SCa, MNCCa/SCa and PMNCa/SCa of the patients with decreased SMg were lower than those of control group. The SMg, MNCMg, PMNMg and RMg were correlated directly with the level of serum albumin.

CONCLUSIONThere are calcium and magnesium deficiencies in the patients with uncompensative cirrhosis and compensative cirrhosis, this deficiency aggravates with the severity of the disease. There is relative increase of intracellular calcium. The magnesium deficiency may be one of the reasons for both hepatic encephalopathy and relative increase of intracellular calcium.

Adult ; Blood Cells ; chemistry ; Calcium ; blood ; Female ; Humans ; Liver Cirrhosis ; blood ; Magnesium ; blood ; Male ; Middle Aged

Gastroenteropancreatic neuroendocrine neoplam (GEP-NEN) is a rare group of tumors with its incidence rising significantly in recent decades. Because of the late presentation of the disease and limitations in conventional biomarkers, about 50% of GEP-NEN patients manifests advanced disease when diagnosed. Therefore, it is vital to identify circulating biomarkers which can not only be used for early diagnosis but also accurately evaluating the biological behavior of GEP-NEN. This review summarizes the advances of circulating biomarkers in diagnosing and evaluating efficacy of treatment in GEP-NEN. Well-known circulating biomarkers include chromogranin A (CgA), pancreastatin (PST), chromogranin B (CgB), neuron-specific enolase (NSE) and pancreatic peptide(PP). Novel biomarkers including circulating tumor cell(CTC), microRNA and NETest are promising biomarkers with potential clinical benefit, but further researches are needed before their clinical applications.
Biomarkers, Tumor ; blood ; Chromogranin A ; blood ; Chromogranin B ; blood ; chemistry ; Gastrointestinal Neoplasms ; blood ; chemistry ; diagnosis ; genetics ; Humans ; MicroRNAs ; blood ; Neoplastic Cells, Circulating ; Neuroendocrine Tumors ; blood ; chemistry ; diagnosis ; genetics ; Pancreatic Neoplasms ; blood ; chemistry ; diagnosis ; genetics ; Pancreatic Polypeptide ; blood ; Phosphopyruvate Hydratase ; blood

BACKGROUND: Becton Dickinson (BD) Vacutainer tubes are the most widely used vacuum system for collection of blood in clinical laboratories. We compared the performance of two new tubes, Sekisui INSEPACK tube and Green Cross Green Vac-Tube, with the existing BD Vacutainer tubes for 49 common analytes. METHODS: A total of 20 apparently healthy volunteers were recruited for this study. For rountine chemistry and thyroid function tests, we compared the results of two new vacutainer tubes and BD Vacutainer tubes with those of BD glass tubes at t =0 hr by student paired t test. Hematology and coagulation test results of the two new vacutainer tubes were compared with those of BD Vacutainer tubes at t =0 hr. To study the stability of each analyte, results at t =24 +/- 2 hr, t =72 +/- 2 hr, and t =168 +/- 2 hr were compared with those at t =0 hr for each tube. RESULTS: Although paired t test analysis revealed statistically significant differences between two tested tubes and existing BD Vacutainer tube in several tests (total protein, total bilirubin, alkaline phosphatase, glucose, uric acid, calcium, inorganic phosphorus, direct bilirubin, triglyceride, HDL-cholesterol, LDL-cholesterol, lactate dehydrogenase, triiodothyronine, and thyroxine), these differences were not considered clinically significant. Stability of two new vacuum tubes for each analyte was similar to that of the BD Vacutainer tube. CONCLUSIONS: Sekisui INSEPACK tube and Green Cross Green-Vac Tube showed a satisfactory analytical performance compared with existing BD Vacutainer tubes. We conclude that these two new plastic vacutainer tubes are acceptable for the commonly ordered laboratory tests.
Blood Cells/chemistry ; Blood Proteins/analysis ; Blood Specimen Collection/*instrumentation ; Equipment and Supplies/standards ; Female ; Humans ; Male

To investigate whether human umbilical cord plasma (HUP) can be used to culture human cord blood mesenchymal stem cells (HUCMSCs), we collected 20 surplus HUP. After being treated with salting out and diasysis, the HUP were used to culture HUCMSCs as 10% volume, and compared with fetal bovine serum (FBS). Morphological characteristics, growth curve and reproductive activity of HUCMSCs cells were observed. The concentration of bFGF and noggin secreted by HUCMSCs cultured with HUP and FBS medium were detected by ELISA. It was found that compared to FBS, the morphology, reproductive activity and characteristic of HUCMSCs cell cultured with HUP were not distinctively different from FBS. The concentration of bFGF in HUP group was significantly higher than that of FBS group, and the concentration of noggin was also different in the two groups. So we concluded that HUP could be used to culture HUCMSCs for a long-time, and the HUP mediumcoild could be more suitable for the culture of human embryonic stem cell (hESC).
Animals ; Cattle ; Cell Culture Techniques ; Cells, Cultured ; Culture Media ; chemistry ; Fetal Blood ; chemistry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Plasma ; chemistry ; Serum ; chemistry

BACKGROUNDHepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide. In order to investigate the molecular biologic mechanism of HCC's development, we studied the expressions of SE-1, CD105 and CD31 in tumor endothelial cells (TECs) of HCC and in the serum of rats.

METHODSWe analyzed the expressions of SE-1, CD31 and CD105 in rat HCC tumor tissues using immunohistochemistry (IHC). Twenty HCC bearing rats and eighteen normal rats were examined for the expressions of SE-1, CD31 and CD105 antigens in serum by enzyme-linked immunosorbent assay (ELISA).

RESULTSSE-1, CD31 and CD105 antigens were detected both in HCC tissue and in normal liver tissue with higher expressions of CD31 and CD105 in HCC while the SE-1 antigen expression was higher in normal liver. Similarly, serum CD31 and CD105 in rats with HCC were significantly increased compared with normal rats (t = 2.8628, P = 0.0086; t = 4.4922, P < 0.0001, respectively). In contrast, SE-1 antigen in HCC rat serum was significantly decreased compared with normal rats (t = 3.4983, P = 0.0011).

CONCLUSIONSE-1, CD31 and CD105 are closely related with liver tumor angiogenesis, which is similar to their performances in terms of their expressions in the serum.

Animals ; Antigens, CD ; blood ; Carcinoma, Hepatocellular ; blood supply ; chemistry ; Endothelial Cells ; chemistry ; immunology ; Enzyme-Linked Immunosorbent Assay ; Immunohistochemistry ; Liver Neoplasms, Experimental ; blood supply ; chemistry ; Male ; Neovascularization, Pathologic ; blood ; Platelet Endothelial Cell Adhesion Molecule-1 ; blood ; Rats ; Rats, Inbred BUF

BACKGROUND: Angiogenin, a 14.1-kDa protein isolated from the medium conditioned by HT-29 human colon carcinoma cells, induces the angiogenesis. In contrast to the human angiogenin, thought to have one homologue, the mouse angiogenin is known to have several angiogenin homologues. During we were investigating the target genes, overexpressed by the chimeric PAX3/FKHR transcription factor, new gene closely similar to the mouse angiogenin rather than angiogenin itself was obtained. We report this Ang-vl gene to understand the process of angiogenesis by comparing the mouse angiogenin family genes. METHODS: The representational difference analysis was used to investigate the target genes over expressed by the PAX3/FKHR chimeric transcription factor. The target genes were subcloned into the pBluescriptSK + and sequenced using the 73 and 77 vector itself primers. Analyses of the completed consensus nucleic acid and peptide sequences were performed using the intelligenetics and GCG software packages as well as BLAST algorithms. RESULTS: The Ang-vl gene, including the glutamine that becomes the N-terminal amino acid by the post-translational peptidase reaction and stop codon, was obtained. CONCLUSIONS: We cloned the one member of the mouse angiogenin family genes. From the point of protein chemistry, the mechanism of angiogenin or, for that matter, of any other blood vessel inducing proteins is not yet known. However, the homologues of the angiogenin might interact each other to regulate the angiogenesls. In this regard, the Ang-vl gene provides an opportunity to understand the mechanism of angiogenesis.
Animals ; Blood Vessels ; Chemistry ; Clone Cells ; Codon, Terminator ; Colon ; Consensus ; Glutamine ; Humans ; Mice* ; Transcription Factors*

Benign symmetric lipomatosis (BSL), also called Madelung's disease, is a rare disease in middle-aged chronic alcohol user. The cause of BSL is unknown. A 63 year-old man with rapid growing lesions in both shoulders for 2 months visited our hospital. Except for cosmetic problem, no abnormal finding was found in blood cell analysis and chemistry; however, excessive fat deposition was found on radiographic findings. Lipoma was revealed in pathologic examination and BSL was diagnosed clinically. Patient is being followed up without any specific problem.
Blood Cells ; Chemistry ; Humans ; Lipoma ; Lipomatosis* ; Lipomatosis, Multiple Symmetrical ; Middle Aged ; Rare Diseases ; Shoulder ; Thoracic Neoplasms

Samples of healthy and full-term human umbilical cord blood samples were obtained asceptically. Mesenchymal stem cells (MSCs) were isolated by lymphocyte separation medium, and were characterized morphologically by fluorescence-activated cell sorting analysis. Differentiation of chondroblast and osteoblast was induced by 10 ng/ml TGF-beta, 100 ng/ml insulin and 10(-7) mol/L decaesadril, 6.25 microg/ml siderophilin, 10 mmol/L beta-sodium glycerophosphate, 50 microg/ml antiscorbic acid, respectirely; the aim was to investigate the potentiality of differentiation. Umbilical cord blood-derived MSCs were stained positive for MSCs marker CD13, CD90, CD166, CD73, CD44 and HLA-AB, but were negative for hematopoietic stem cell marker CD45, CD34 and HLA-DR. After 21 days induction, Toluidine Blue staining and von-Kossa staining were positive. Immunocytochemistry showed that Collagen II expressed in the induced cells. The results demonstrated that mesenchymal stem cells can be isolated from human umbilical cord blood and differentiated into chondroblasts and osteoblasts in vitro.
Cell Differentiation ; Cell Separation ; Cells, Cultured ; Chondrocytes ; cytology ; Fetal Blood ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; chemistry

OBJECTIVEThe study was to investigate the impact of cord blood CD(3)AK cell culture supernatant (CS) on proliferation, differentiation and apoptosis of HL-60 cells.

METHODSHL-60 cells were treated with different concentrations of CS (10%, 15%, 20%) for 3 days, 6 days and 9 days, and the same cells of control group were not treated with CS. The growth of induced cells was assessed with Trypan blue staining and cell counting with cytometer. The differentiation marker CD(11b) on the cell surface and cell-cycle was analyzed by flow cytometry (FCM), cell morphology (Wright-Giemsa staining) and NBT test to determine the extent of differentiation. Meanwhile, the changes of the apoptosis of the cells induced by 20% CS at different time points (3, 6 and 9 days) were analyzed by TUNNEL-POD, and the apoptotic characteristics of cells were observed.

RESULTSThe growth of HL-60 cell was inhibited as CS-inducing time and the dose of CS increased. At the same time, but HL-60 cell number in G(0)/G(1) phase of cell-cycle increased, but HL-60 cell number in S phase decreased compared with untreated group. The HL-60 cells induced by 20% CS for 9 days showed that (52.7 +/- 1.8)% of cells were at G(0)+G(1) phase and (43.8 +/- 1.1)% were at S phase (P < 0.05), which demonstrated that HL-60 cells induced by 20% CS underwent G(0)/G(1) phase cell-cycle arrest. The volume of the differentiated cells was enlarged gradually as CS-inducing time prolonged. After 3 days the differentiating cells began to express differentiating marker CD(11b) on the cell surface and the nuclei morphology of the differentiated cells was also changed and NBT-stained cells increased in number with the increased dose of CS increased. Three days after induction by 20% CS, the induced cells began to show signs of apoptosis and the apoptotic percentage of induced cells gradually increased with CS-induction time. The rate of apoptosis of cells was (33.3 +/- 2.3)% at 9 days (P < 0.01).

CONCLUSIONCS could not only inhibit the growth of HL-60 cells but also induce the differentiation and apoptosis in HL-60 cells.

Apoptosis ; Cell Culture Techniques ; Cell Cycle ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Culture Media ; chemistry ; Fetal Blood ; chemistry ; HL-60 Cells ; Humans

This study was aimed to look for the potential application of human umbilical plasma (HUP) in the culture of human umbilical vein endothelial cells (HUVECs). The effect of HUP concentration in cell culture medium on cell proliferation activity and cell cycle was studied. HUVECs were obtained by digesting the umbilical cord with 0.25% trypsin mixed with the equal volume of 0.1% collagenase II, then were identified by morphology and factor VIII immunohistochemistry under phase contrast microscopy. The cells cultured after 7 days showed the typical cobblestone morphology with factor VIII immunohistochemical staining positive. The study showed that the groups of 20% (HUP), 15% HUP+20% Fetal Bovine Serum (FBS) and 20% HUP+20%FBS enhanced cell proliferation activity significantly when compared with the control group (20% FBS without HUP). On the contrary, 30% HUP+20% FBS caused cell cycle arrest, which significantly hindered the proliferation of HUVECs. The study proved that although HUP might not be able to completely replace the role of vascular endothelial growth factor in cell culture, as a supplement ingredient, it was an ideal candidate to replace FBS in culture medium.
Cell Culture Techniques ; methods ; Cell Proliferation ; Cells, Cultured ; Culture Media ; chemistry ; Fetal Blood ; chemistry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Plasma ; physiology