Background Little information is available regarding the effect of angiotensin Ⅱ (Ang Ⅱ) on the bumetanide-sensitive sodium-potassium-2 chloride cotransporter (NKCC2), the thiazide-sensitive sodium-chloride cotransporter (NCC), and the Cl- channel (CLC)-K2 at both mRNA and protein expression level in Ang Ⅱ-induced hypertensive rats. This study was conducted to investigate the influence of Ang Ⅱ with chronic subpressor infusion on nephron-specific gene expression of NKCC2, NCC and CLC-K2. Results Ang Ⅱ significantly increased blood pressure and up-regulated NKCC2 mRNA and protein expression in the kidney. Expression of CLC-K2 mRNA in the kidney increased 1.6 fold (P<0.05).There were no changes in NCC mRNA or protein expression in AngII-treated rats versus control. Conclusions Chronic subpressor Ang Ⅱ infusion can significantly alter NKCC2 and CLC-K2 mRNA expression in the kidney, and protein abundance of NKCC2 in kidney is positively regulated by Ang Ⅱ. These effects may contribute to enhanced renal Na+ and Cl- reabsorption in response to Ang Ⅱ.

Objective: To observe the effect of emodin on the biological behavior of human fibroblasts (FB) in culture of kidney in patients with lupus nephritis (LN). Methods: FB were isolated from kidney culture of LN patients, and the effect of emodin on 3　H-TdR incorporated rate of FB was observed. The apoptosis and c-myc gene expression were detected in the same way by flow cytometry. Results: Emodin could markedly inhibit the proliferation of human kidney FB, and inducing cell apoptosis through up-regulating c-myc gene expression in human renal FB. Conclusion: Emodin can inhibit proliferation and promote apoptosis of FB, which may be important in ameliorating interstitial fibrosis, and thus improve prognosis of LN.

Objective:To explore the changes of iron metabolism in patients with hepatitis B cirrhosis and esophageal and gastric varices complicated with portal vein thrombosis.Methods:This study was a cross-sectional study. 253 patients with hepatitis B cirrhosis with esophageal and gastric varices who were hospitalized in the Zhongshan Hospital, Fudan University from January 1, 2020 to December 31, 2021 were included in this study. They were divided into portal vein thrombosis group ( n=57) and non portal vein thrombosis group ( n=196) according to the presence or absence of portal vein thrombosis. The iron metabolism characteristics of the two groups were compared, and subgroups were analyzed according to the presence or absence of ascites, platelet count level, D-dimer level, and Child grade. The factors related to portal vein thrombosis were screened through multivariate logistic regression analysis. Results:The ratio of Child pugh B/C, ascites, D-dimer and platelet count in patients with hepatitis B cirrhosis and esophageal and gastric varices complicated with portal vein thrombosis was higher (all P<0.05). Patients with portal vein thrombosis had higher levels of soluble transferrin receptor [2.4(1.8, 3.6)mg/L vs 1.8(1.3, 2.7)mg/L, P=0.006], and lower levels of ferritin [33.1(18.9, 63.3)ng/ml vs 57.7(19.4, 142.5)ng/ml, P=0.038]. Layered analysis showed that ascites, platelet count levels, D-dimer levels, and Child-pugh grade did not affect the negative correlation trend between ferritin and portal vein thrombosis, and the positive correlation trend between soluble transferrin receptors and portal vein thrombosis. Moreover, soluble transferrin receptors showed a statistically significant positive correlation with portal vein thrombosis in the absence of ascites, low D-dimer levels, and Child-pugh grade A. Multivariate analysis suggested that after weighing Child-pugh grading, platelet count, and D-dimer levels, ferritin ( OR=0.943, 95% CI: 0.904-0.983, P=0.006) and soluble transferrin receptor ( OR=1.034, 95% CI: 0.001-1.067, P=0.044) were independently associated with portal vein thrombosis. Conclusions:In patients with hepatitis B cirrhosis with esophageal and gastric varices, the characteristics of iron metabolism in patients with portal vein thrombosis are different from those in patients without thrombosis, with higher levels of soluble transferrin receptor and lower levels of ferritin.

Animals ; DNA, Complementary ; immunology ; Dendritic Cells ; immunology ; Female ; Liver Neoplasms, Experimental ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; alpha-Fetoproteins ; immunology

OBJECTIVETo establish an inductively coupled plasma mass spectrometry(ICP-MS) method for determination of 30 trace elements including As, Ba, Be, Bi, Ni, Cd, Co, Cr, Cs, Cu, Ga, Mn, Pb, Sr, Tl, V, Ge, Mo, Nb, Ti, W, Te, Se, Zr, In, Sb, Hg, Ce, La, and Sm in human blood.

METHODThe blood samples were analyzed by ICP-MS after diluted 1/10 with 0.01% Triton-X-100 and 0.5% nitric acid solution. Y, Rh and Lu were selected as internal standard in order to correct the matrix interference of Cr, As, Se, and Hg by a hex pole-based collision-reaction cell. Other elements were determined with standard method. The limits of detection, precision and accuracy of the method were evaluated. The accuracy was validated by the determination of the whole blood reference material.

RESULTSAll the 30 trace elements have good linearity in their determination range, with the correlation coefficient > 0.9999. The limits of detection of the 30 trace elements were in the range of 1.19 - 2.15 µg/L and the intra-precision and inter-precision (relative standard deviation, RSD) were less than 14.3% (except Hg RSD < 21.2%, and Ni RSD < 15.4%). The spiked recovery for all elements fell within 59.3% - 119.2%. Among the 13 whole blood reference materials, V, Cr, Mn, Co, Ni, Cu, As, Se, Cd, Te, and Pb (1.45, 1.19, 18.40, 0.18, 1.57, 591.00, 2.97, 61.00, 0.35, 1.86, and 9.70 µg/L respectively) fell within the acceptable range and the detection results of Hg (0.59 µg/L) and Mo (1.59 µg/L) were slightly beyond the range.

CONCLUSIONThis method was simple, fast and effective. It can be used to monitor the multi-elementary concentration in human blood.

Humans ; Limit of Detection ; Mass Spectrometry ; methods ; Metalloids ; blood ; Metals ; blood ; Trace Elements ; blood

BACKGROUNDLittle information is available regarding the effect of angiotensin II (Ang II) on the bumetanide-sensitive sodium-potassium-2 chloride cotransporter (NKCC2), the thiazide-sensitive sodium-chloride cotransporter (NCC), and the Cl- channel (CLC)-K2 at both mRNA and protein expression level in Ang II-induced hypertensive rats. This study was conducted to investigate the influence of Ang II with chronic subpressor infusion on nephron-specific gene expression of NKCC2, NCC and CLC-K2.

METHODSSprague Dawleys rats were treated subcutaneously with either Ang II (100 ng.kg-1.min-1) or vehicle for 14 days. Expression of NKCC2, NCC and CLC-K2 mRNA in kidneys was determined by real time polymerase chain reaction (PCR). Western blotting analysis was used to measure NKCC2 and NCC protein expression.

RESULTSAng II significantly increased blood pressure and up-regulated NKCC2 mRNA and protein expression in the kidney. Expression of CLC-K2 mRNA in the kidney increased 1.6 fold (P < 0.05). There were no changes in NCC mRNA or protein expression in AngII-treated rats versus control.

CONCLUSIONSChronic subpressor Ang II infusion can significantly alter NKCC2 and CLC-K2 mRNA expression in the kidney, and protein abundance of NKCC2 in kidney is positively regulated by Ang II. These effects may contribute to enhanced renal Na+ and Cl- reabsorption in response to Ang II.

Angiotensin II ; pharmacology ; Animals ; Blood Pressure ; drug effects ; Gene Expression Regulation ; drug effects ; Hypertension ; chemically induced ; metabolism ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Chloride Symporters ; genetics ; Solute Carrier Family 12, Member 1

The study was aimed to explore the relationship between patterns of methylation or deletion and the development of acute leukemia, and further to clarify the possible mechanism in the development of adult acute leukemia. Nested methylation-specific polymerase chain reaction (n-MSP) was adopted to analyze p16 gene methylation or deletion patterns in 82 adult acute leukemia patients with different subtypes and stages. The results indicated that rate of p16 gene methylation was 39.0% in 82 adult acute leukemia patients, among them, 41.4% in acute myelogenous leukemia (AML) and 33.3% in acute lymphoblastic leukemia (ALL). It were found that 36.6% of de novo AL patients and 54.5% of relapsed AL patients developed the hypermethylation of p16 gene. Out of the 82 patients, 6 seemed to have deletion of p16 gene, including 1 AML (1.7%) and 5 ALL (20.8%). There were no hypermethylation or deletion of p16 gene in the 16 controls. It is concluded that methylation of p16 gene may play a more important role than homozygous deletion of p16 gene in the leukemogenesis and progression of adult acute leukemia.
Adolescent ; Adult ; Aged ; Base Sequence ; CpG Islands ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Female ; Genes, p16 ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

OBJECTIVETo explore the distribution of genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) 677C/T, 1298A/C and methionine synthase reductase (MTRR) 66A/G among ethnic Han females from Linyi, and to correlate it with serum level of homocysteine (Hcy).

METHODSA cross-sectional study was carried out. Oral epithelial cell samples were collected from 825 subjects. MTHFR and MTRR gene polymorphisms were determined with a Taqman-Minor Groove Binder (MGB) method. Distribution of gene polymorphisms was analyzed and compared with others regions of China including Weifang, Zhengzhou, Deyang and Hainan. A biochemical assay was also carried out to determine the total Hcy in plasma of 281 subjects. The reductase activity of MTHFR was classified into decreased and stable groups according to genetic polymorphism of MTHFR. Correlation between MTHFR groups and total Hcy level were also explored.

RESULTS(1) The frequencies of MTHFR677CC, CT and TT genotypes of the selected subjects were 16.7%, 48.3% and 35.0%, respectively. The frequencies of MTHFR 1298AA, AC and CC genotypes were 76.0%, 21.6% and 2.4%, respectively. And those of MTRR 66AA, AG and GG genotypes were 54.7%, 39.4% and 5.9%, respectively. For the selected subjects, their frequency of MTHFR 677TT genotype was higher than that of Deyang and Hainan (P< 0.01), whilst the frequency of MTHFR 1298CC genotype was lower than that of Deyang and Hainan (P < 0.01), and the frequency of MTRR 66 GG genotype was lower than that of Hainan (P< 0.01). (2) The Hcy level for those with decreased MTHFR activity was significantly higher than those with stable MTHFR activity (P< 0.05).

CONCLUSIONMTHFR gene 677C/T, 1298A/C and MTRR 66A/G polymorphisms in ethnic Han women from Linyi have differed significantly from other regions of China. Decreased MTHFR activity caused by genetic polymorphisms is a risk factor for raised Hcy level.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Ferredoxin-NADP Reductase ; genetics ; Gene Frequency ; Genetic Association Studies ; Genotype ; Homocysteine ; blood ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; blood ; genetics ; Polymorphism, Single Nucleotide ; Young Adult

To investigate the effects of matrine on apoptosis and expression of adhesion molecules in human multiple myeloma cell line RPMI8226 cells, RPMI8226 cells were incubated with indicated concentrations of matrine. The growth of RPMI8226 cells was observed by CCK-8 colorimetric assay and apoptosis was detected by flow cytometry using Annexin V-FITC/PI staining. The cell cycles were analyzed by PI staining. Flow cytometry using Annexin V-FITC/PI staining was used to detect the expression of cell adhesion molecules, including CD44, CD44v6, CD54 and CD106. The results showed that RPMI8226 cell viability in presence of matrine decreased markedly in a dose- and time-dependent manners. The apoptosis could be induced by matrine and its level increased following the augmentation of the drug concentration. After treated by matrine for 48 hours, a concentration-dependent increase of cells in G(0)/G(1) phase and a decrease in S phase could be detected, but no obvious change of cell count was found in G(2)/M phase. Treatment of RPMI8226 cells with matrine for 48 hours resulted in decrease of expression levels of CD44 and CD54, while expressions of CD44v6 and CD106 had no significant change. It is concluded that matrine induces in vitro apoptosis, suppresses proliferation in multiple myeloma cells and depresses expression of some adhesion molecules.
Alkaloids ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Hyaluronan Receptors ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Multiple Myeloma ; pathology ; Quinolizines ; pharmacology

To explore the hematopoietic reconstitution and transplantation-related complications of two units of unrelated umbilical cord blood combined transplantation for the treatment of adult hematologic malignancies, one adult patient with chronic myelogenous leukemia received two units of unrelated umbilical cord blood combined transplantation. The conditioning regimen was busulfan and cyclophosphamide (Bu-Cy). GVHD prophylaxis regimen consisted of mycophenolate mofetil (MMF), cyclosporine A (CsA) and methotrexate (MTX). The patient received total nucleated cells 4.63 x 10(7)/kg with CD34+ cells 8.34 x 10(5)/kg. Engraftment was documented by the analysis of short tandem repeat with polymerase chain reaction (STR-PCR). The results showed that the STR-PCR analysis for peripheral blood at day 31, 46 and 71 after transplantation suggested that one of two units of cord blood were completely engrafted. The ANC > 0.5 x 10(9)/L in the patient occurred at day 23, blood platelet counts > 20 x 10(9)/L at day 33 and > 50 x 10(9)/L at day 47. The Philadelphia chromosome and bcr/abl fusion gene of the patient also turned to negative after engraftment. Acute GVHD grade II occurred at day 13 and cured after treatment. It is concluded that umbilical cord blood can be used in adult hematopoietic stem cell transplantation. Two or more units umbilical cord blood combined transplantation might be the way to solve the problem of the low counts of nucleated cells when be used for adult.
Adult ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; therapy ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; therapy ; Leukocyte Count