Objective To investigate the effects of female sex hormones in cow's milk on metabolism of blood lipid in young male rats.Methods Forty-eight male Sprague-Dawley rats aged 21 days old were assigned randomly to 4 groups,each containing 12 rats,and fed with quantitative milk from postpartum cow,milk from pregnant cow,commercial whole milk and artificial milk,respectively.Serum total cholesterol (TC),triacylglyeriol(TG),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C) and urinary creatinine (Cr) were determined with automatic biochemical analyzer.Serum progesterone(P4)and urinary free estriol(UFE3) were determined with immunochemiluminometric assays after all rats were killed at 53 days old.SPSS 13.0 software was used to analyze the data.Results Total estradiol and P4 were 1 189.66 pmol/L,833.98 pmol/L,588.17 pmol/L,286.48 pmol/L and 9.76 nmol/L,10.18 nmol/L,2.83 nmol/L,0.92 nmol/L in milk from pregnant cow,commercial whole milk,milk from postpartum cow and artificial milk groups,respectively.Serum TC were respectively(1.78?0.29) mmol/L,(1.94?0.20) mmol/L,(2.10?0.28) mmol/L and (2.11?0.22) mmol/L in pregnant milk,commercial whole milk,postpartum milk and artificial milk groups,and TC in pregnant milk group was lower than that in postpartum milk group or artificial milk group(P0.05).Conclusion Milk from pregnant cow may reduce serum TC in young male SD rats,which may be related to the conjoined effect of estradiol and P4.

OBJECTIVETo observe and compare the luciferase activities of different length segments of human dentin matrix protein 1 promoter in human dental pulp stem cells (HDPSC), osteoblasts (OC) and Hela cells.

METHODSThe differentlength desired DNA segments were obtained from 2 195 bp Dmp1 promoter cloned by PCR method. The amplified promoter segments with different length were cloned into luciferase report gene vector pGL3-Basic, the correct orientation of those inserts was verified by cutting with two different restrict enzymes. The luciferase activity was observed after different pGL3-PDmp1 vectors were transfected transiently into those three different-type cells.

RESULTS6 Dmp1 promoter segments with different-length were obtained successfully, and luciferase report gene vectors with different promoter segments were successfully constructed after identified by restriction enzymes cutting. They had different luciferase activities when they were transfected transiently into HDPSC, and the region of -505(-)-193 bp and -935(-)-505 bp could be regarded as the specific promoters of Dmp1 promoter for HDPSC and OC respectively, which could include the basic regulatory elements.

CONCLUSIONThe correct clone of the upstream of human Dmp1 promoter segments with different length had been obtained, and they had strong luciferase activities in HDPSC and OC, but very low in Hela cell. These results will make an important basis for studying mineralized tissue-specific transcriptional regulation mechanisms of Dmp1.

Dentin ; Extracellular Matrix Proteins ; Gene Expression Regulation ; Genetic Vectors ; Humans ; Phosphoproteins ; Promoter Regions, Genetic ; Transfection

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism

This study was purposed to explore the changes of possible angiogenetic factors other than VEGF after inhibition of NHE1 and their related mechanisms. The K562 cells were treated by NHE1 specific inhibitor cariporide, the angiogenesis factors after inhibition of NHE1 were screened by using protein chip, the IL-8 expression level after cariporide treatment was detected by real-time quantitative PCR; the K562 cells with stable interference of NHE1 were constructed, the IL-8 expression level after interference of NHE1 was detected by real-time quantitative PCR; the p38 phosphorylation level in K562 cells treated with cariporide was detected by Western blot. After treatment of K562 cells with p38 inhibitor SB203580, the IL-8 expression level was decreased by real-time quantitative PCR. The results of protein chip showed that IL-8 expression decreased after cariporide treatment. Real-time quantitative PCR confirmed this inhibitory effect. The p38 phosphorylation level increased after cariporide treatment. The down-regulation of IL-8 expression induced by cariporide treatment was partially restored after K562 cells were treated with p38 inhibitor SB203580. It is concluded that the inhibition of NHE1 can inhibit IL-8 expression through up-regulation of p38 phosphorylation.
Cation Transport Proteins ; antagonists & inhibitors ; Down-Regulation ; Guanidines ; pharmacology ; Humans ; Imidazoles ; pharmacology ; Interleukin-8 ; metabolism ; K562 Cells ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; Sulfones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

This study was aimed to investigate the expression of RHBDD1 gene in patients with chronic myeloid leukemia (CML) and explore its clinical significance. The relative expression levels of RHBDD1 in bone marrow mononuclear cells of healthy controls and CML patients were detected by using real time PCR. The results showed that the expression level of RHBDD1 in CML patients was significantly higher than that in healthy controls. The expression level of RHBDD1 in CML patients with negative BCR/ABL p210 was remarkably higher than that in patients with positive BCR/ABL p210. In patients ≥ 50 years old RHBDD1 expression was lower than the patients < 50 years old. There were no significant relation of RHBDD1 expression with sex of patients. It is concluded that RHBDD1 gene may be involved in the pathogenesis and progression of CML, particularly reflects in the pathogenesis of the patients with negative BCR/ABL p210.
Bone Marrow Cells ; metabolism ; pathology ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Serine Endopeptidases ; genetics ; metabolism

This study was aimed to investigate whether the inhibition of NHE1 activity and intracellular acidification can reverse resistance of leukemia cells to the imatinib and to explore downstream signal molecule networks of BCR/ABL in the cells of chronic myelocytic leukemia (CML) patients. The mRNA and protein expression of P-glycoprotein (Pgp) and the drug accumulation were assayed after acidifying the primary leukemia cells of patients or K562/DOX and K562/G01 cells. The effects of intracellular acidification of primary leukemia cells on the phosphorylation level changes of ERK1/2 and p38 MAPK were analyzed by Western blot. The results showed that the intracellular concentration of drugs in the advanced patients increased and the sensitivity of K562/DOX and K562/G01 cells to imatinib was enhanced after intracellular acidification or treatment with NHE1 inhibitor cariporide. With downregulation of intracellular pH, the phosphorylation of p38 MAPK decreased in advanced patients and the phosphorylation of ERK1/2 increased within 3 min and then decreased after 30 min. SB203580, the specific inhibitor of p38 MAPK, displayed a synergistic effect with the inhibitor of NHE1 to downregulate the mRNA and protein expression of Pgp. It is concluded that the inhibiton of NHE1 can significantly decrease the protein expression of Pgp in K562/DOX and K562/G01 cells, increase the accumulation of Rhodamine123 and doxorubicin in the cells of advanced patients and enhance the sensitivity of cells to imatinib in which the p38 MAPK signal transduction pathways involves.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Benzamides ; pharmacology ; Cation Transport Proteins ; antagonists & inhibitors ; metabolism ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Inhibitors ; pharmacology ; Gene Expression Regulation, Leukemic ; Humans ; Imatinib Mesylate ; Imidazoles ; pharmacology ; K562 Cells ; MAP Kinase Signaling System ; Piperazines ; pharmacology ; Pyridines ; pharmacology ; Pyrimidines ; pharmacology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; metabolism ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo evaluate the main performance of BS-200 automatic biochemistry analyzer.

METHODSTo evaluate the repeatability, linearity, precision and accurate Of the BS-200 automatic biochemistry analyzer by 8 items that contain all wavelength (ALT, GGT, Cl-, Cr, Glu, TP, Alb. APOA1. TBil), and carry on the contrast experiment with Hitachi 7170.

RESULTSThe daytime run CV,within-run CV between-run CV and total CV are under 4.0%, the duplication is well, the accuracy is high, the linear scope is wide, and the contrast experiment correlation is good.

CONCLUSIONThis instrument each appraisal target is good, and worth in the small or middle-sized hospital promotion application.

Automation ; instrumentation ; Biochemistry ; instrumentation ; Reproducibility of Results

OBJECTIVETo observe the clinical effect of Dihuang Zhixue Capsule (DZC, a Chinese preparation for cooling blood and dispelling toxic substances) in the treatment of childhood refractory idiopathic thrombocytopenic purpura (RITP), with cyclosporin A (CsA) used as the control.

METHODSForty-one children of RITP were randomized into the treated group and the control group. The 21 patients in the treated group were orally given 2 to 3 DZC capsules each time, thrice a day and the 20 in the control group were given 3 mg/kg CsA per day, with 3 months as one therapeutic course. The therapeutic efficacy, platelet count and adverse reaction in the two groups were compared at the end of the course.

RESULTS(1) In the treated group, 1 (4.8%) patient was evaluated as cured, 3 (14.3%) as markedly effective, 5 (23.8%) as effective, 5 (23.8%) as improved, 7 (33.3%) as ineffective, with the total effective rate being 66.7%; while in the control group, the corresponding numbers were 0, 2 (10.0%), 2 (10.0%), 3 (15.0%), 13 (65.0%) and 35.0%, respectively, showing statistical significance in difference between the total effective rates of the two groups (xi(2)=4.11, P=0.0426). (2) As compared with the baseline, the platelet count increased in both groups after 2 months' treatment (P<0.05). After 3 months' treatment, the platelet count was higher in the treated group than in the control group (P<0.05). (3) The improvement of hemorrhage in the treated group after 8 weeks' treatment was better than that in the control group (P<0.05). (4) No apparent adverse reaction was observed in the treated group, while in the control group, hirsutism was shown in 15 cases; gingival hyperplasia in 10; digestive reaction in 5, liver function impairment in 5, hypertension in 2 and renal impairment in 2.

CONCLUSIONThe therapeutic efficacy of DZC is better than that of CsA, and DZC shows good compliance but brings no obvious adverse reaction.

Adolescent ; Capsules ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; Humans ; Male ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; Treatment Outcome

This study was aimed to investigate the expression level of CIAPIN1 mRNA in leukemia patients and explore its significance in leukemias. The fresh bone marrow was collected from 112 newly diagnosed leukemia patients, the total RNA was extracted by means of TRIzoL, the cDNA was synthesized, the expression of CIAPIN1 mRNA was detected by real-time quantitative PCR using β-actin as internal reference; 10 normal healthy persons were selected as controls. The results showed that the expression of CIAPIN1 mRNA was statistically higher in acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and chronic phase chronic myeloid leukemia (CML) patients than that in normal persons (p < 0.05); but there was no statistical difference between chronic lymphocytic leukemia (CLL) and normal persons (p > 0.05). It is concluded that the CIAPIN1 gene higher expresses in MNC of newly diagnosed leukemia patients, up-regulation of CIAPIN1 expression may play an important role in pathogenesis of leukemia.
Adult ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Leukemia ; genetics ; metabolism ; pathology ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the impact of intracellular acidification (IA) on drug resistance of leukemia cells with high P-glycoprotein (P-gp) expression, and to provide a new method for the reversing of multidrug resistance (MDR).

METHODSReal-time PCR was used to determine the expression level of mdr1 gene, and the leukemia cells with high P-gp expression were selected. The specific inhibitor of Na+/H+ exchanger 1 and the "high K+" buffer were used to acidify the cells, and the confocal laser microscopy was used to determine the intracellular pH (pHi) and effect of IA on the accumulation of doxorubicin. The MTT method was used to determine the effect of IA on the cell viability. The flow cytometry was used to detect the effect of IA on the P-gp function, and Western blotting was used to determine the effect of IA on the expression of P-gp.

RESULTSThe pHi was decreased to 7.0, and compared with that of control the mdr1 mRNA expression was decreased to (53.2+/-11.0)% after 1 h, and to (16.6+/-7.0)% after 3 h treatment. The P-gp expression was decreased to (56.0+/-9.0)% of the control after 3 h treatment. The accumulation of Rh123 was 71.03+/-0.47 at pHi 7.0, which was increased obviously as compared to the control group 20.07+/-0.39. The increased accumulation of doxorubicin was also observed by confocal laser microscopy.

CONCLUSIONThe expression and function of P-gp on the patients cells are inhibited by IA.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Cell Survival ; drug effects ; Doxorubicin ; pharmacokinetics ; Drug Resistance, Neoplasm ; drug effects ; Guanidines ; pharmacology ; Humans ; Hydrogen-Ion Concentration ; drug effects ; Leukemia ; drug therapy ; metabolism ; RNA, Messenger ; genetics ; Sulfones ; pharmacology ; Tumor Cells, Cultured