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1.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-995087

ABSTRACT

Objective:To analyze the prevalence of parturiophobia and its association with preferred mode of delivery in pregnant women in Changning District, Shanghai.Methods:A cross- sectional study was conducted among 1 560 pregnant women in the third trimester who had their antenatal examination in Changning Maternity and Infant Health Hospital from September 2020 to March 2021. Fear of childbirth was measured with a validated Chinese version of Wijma Delivery Expectancy/Experience Questionnaire version A (W-DEQ-A). Based on the W-DEQ-A scores, the participants were divided into two groups: non-clinical parturiophobia group [<85 scores, including mild (≤37 scores), moderate (38-65 scores) and severe (66-84 scores) parturiophobia] and clinical parturiophobia group (≥85 scores). Rank-sum test, Chi-square test and t-test were used for univariate analysis. Multivariate binary logistic regression was used to analyze the factors associated with fear of childbirth and its relationship with preferred mode of delivery. Results:The detection rates of mild, moderate, severe and clinical parturiophobia were 18.8% (294/1 560), 44.9% (700/1 560), 31.1% (485/1 560) and 5.2% (81/1 560), respectively. Multivariate binary logistic regression showed that the participants who were supported by relatives and friends to have cesarean section ( OR=3.45, 95% CI: 1.29-9.22) or had antenatal anxiety ( OR=4.73, 95% CI: 2.49-8.97) were more likely to have clinical parturiophobia, while those with planned pregnancy ( OR=0.49, 95% CI: 0.29-0.82), high intensity physical activity ( OR=0.36, 95% CI: 0.18-0.72) or better/well understanding of the delivery process ( OR=0.42, 95% CI: 0.19-0.97) were less likely to develop clinical parturiophobia (all P<0.05). Compared with the non-clinical parturiophobia women, those with clinical parturiophobia were more likely to choose cesarean section ( OR=2.15, 95% CI: 1.22-3.78, P=0.008). Conclusions:The detection rates of severe and clinical parturiophobia are 31.1% and 5.2% in Changning District, Shanghai. The associated factors mainly include the attitudes of relatives and friends towards the mode of delivery, antenatal anxiety, planned pregnancy or not, physical activity level and the understanding of delivery process. Clinical parturiophobia might be an important factor for cesarean section on maternal request.

2.
Journal of Experimental Hematology ; (6): 1071-1078, 2016.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-246814

ABSTRACT

<p><b>OBJECTIVE</b>To study the relationship between surface markers of CD56 and CD19 and karyotypes and prognosis in multiple myeloma.</p><p><b>METHODS</b>A total of 126 cases of newly diagnosed multiple myeloma in the first hospital of Peking university from 2011 to 2015 were enrolled in this study. Cytogenetic abnormalities and immunophenotypes were detected by using fluorescence in situ hybridization and flow cytometry respectively before chemotherapy. Bone marrow smear was used for detection of abnormal plasma cell infiltration. By combining with their basic data, the relationship between immunophenotypes, cytogenetics and prognosis of MM was analyzed.</p><p><b>RESULTS</b>(1) The median of myeloma cells in the 126 patients was 0.24(0.01-0.97); the median of myeloma cells in 116 patients who have immunophenotype datas was 0.25(0.01-0.97); the median of myeloma cells in CD19 positive patients was 0.11(0.01-0.53); the median of myeloma cells in CD19 negative patients was 0.26(0.01-0.97). The median of myeloma cells in CD19 positive patients was much lower than that in CD19 negative patients(P=0.036). (2)In 116 patients detected by the immunophenotype, the myeloma cells expressed CD19,CD20,CD56 and CD117. Compared with CD56 negative patients(45/116,38.79%),CD56 positive patients(71/116,61.21%) had a clearly favorable disease outcome(OS was 53.0 month vs 31.0 month,P=0.016; PFS was 37.5 months vs 18.4 months, P=0.036). (3)CD19 positive patients was 16.38%(19/116),CD19 negative patients was 83.62%(97/116); CD19 positive MM and CD19 negative MM had no difference in OS and PFS. (4)CD117 positive rate in CD19 positive patients was 42.11%(8/19), the CD117 positive rate in CD19 negative patients was 18.57%(18/97), the CD19 expression positively correlated with CD117 expression. (5)FISH detection was done for 67 newly diagnosed MM patients, 8 patients showed normal karyotypes(11.94%), 59 patients had abnormal karyotypes(88.06%). The most common abnormal karyotypes were IgH rearragement which occurred in 47 patients(70.15%). Other abnormal karyotypes included 1q21+, del(13q14),del(13q14.3),del(17p13) . These abnormal karyotypes occurred in 37 patients(55.22%),31 patients(46.27%),33 patients(49.25%) and 13 patients(19.40%) respectively. In comparison with CD19 negative MM patients, the incidence rate of 1q21+ and del(13q14.3) was significantly lower in CD19 positive patients(1q21+:33.33% vs 61.54%,P=0.016; del(13q14.3): 33.33% vs 53.85%,P=0.043).</p><p><b>CONCLUSION</b>The prognosis of CD56 positive MM patients is better than that of CD56 negative MM patients, CD19 negative MM has more abnormal karyotypes and bone marrow infiltration,but they have no statistical prognostic differences.</p>


Subject(s)
Humans , Chromosome Aberrations , Chromosome Deletion , Flow Cytometry , Immunophenotyping , In Situ Hybridization, Fluorescence , Karyotyping , Multiple Myeloma , Prognosis
3.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-246829

ABSTRACT

<p><b>OBJECTIVE</b>To analyse the clinical features and prognostic significance of cross-lineage antigen expression in patients with acute myeloid leukemia(AML) in order to establish individualized treatment for a better outcome and prognosis.</p><p><b>METHODS</b>A total of 227 cases (exduding M3) were detected by flow cytometry for immune phenotype. The CD7(-)CD56(-)CD19(-) AML served as control. The clinical features, treatment response and prognosis of CD7(+) group, CD56(+) group and CD19(+) group were compared.</p><p><b>RESULTS</b>The detection rate of CD56(+),CD7(+) and CD19(+) in AML was 15.9%, 25.1% and 11.0%, respectively. There were no differences between CD56(+) AML, CD7(+) AML, CD19(+) AML, and CD56(-)CD7(-)CD19(-) AML in the proportion of blast cells, white blood cell count, hemoglobin level, platelet count and MDS transformed AML rate. The CR after the first course chemotherapy and cumulative CR in CD56(+) AML patients were lower than those in the control group (20.0% vs 58.1%, P=0.0099; 73.3% vs 87.5%, P=0.04). The median time of CR in CD56(+) AML was longer than that in the control group (118 days vs 46 days, P=0.04). The PFS time and OS time of CD56(+) AML were shorter than those in the control group (245 days vs 580 days, P=0.037; 494 days vs 809 days, P=0.04). The CR after the first course chemotherapy and cumulative CR in CD19(+) AML patients were higher than those in the control group(75.0% vs 58.1%, P=0.46; 100% vs 87.5%, P=0.02). The median time of CR in CD19(+) AML was shorter than that in the control group (28 days vs 46 days, P=0.02). The PFS time and OS time of CD19(+) AML tended to be longer than those in the control group (P=0.13, P=0.07, respectively). The median PFS and OS were not reached at the time of last follow-up. The CR after the first course chemotherapy, cumulative CR and median time to CR in CD7(+) AML patients were not different from those in the control group (53.1% vs 58.1%, P=0.67; 87.1% vs 87.5%, P=0.44; 50 days vs 46 days, P=0.44). No differences of PFS and OS were observed between CD7(+) AML and the control.</p><p><b>CONCLUSION</b>CD56(+) AML patients respond poorly to treatment, frequently relapse after complete remission and have a low survival rate. These patients need more intensive chemotherapy or in combination with other treatments. The interval of MRD detection should be shortened to find out relapse earlier. CD19(+) AML patients have a good treatment outcome and often accompanies with AML1/ETO fusion gene, which is known to be a good prognostic marker. Aberrant expression of CD7 on AML cells is not a poor prognostic factor in this study.</p>


Subject(s)
Humans , Antigens, CD , Flow Cytometry , Immunophenotyping , Leukemia, Myeloid, Acute , Prognosis , Remission Induction , Survival Rate
4.
Article in Chinese | MEDLINE | ID: mdl-26016229

ABSTRACT

OBJECTIVE: To investigate the effects of Artesunate(Art) on the LX-2 cell. METHODS: The cultured hepatic stellate cells were divided into control group and Art-treated groups with 250,350,450 µmol/L. The rate of cellular proliferation was detected by MIT assay, the content of ceramide (Cer)was determined by HPLC method, the content of hydroxyproline (Hyp) was determined by enzyme digestion method, the expressions of PPAR-γ, p53 and Caspase 3 were detected by Western blot. RESULTS: Compared with control group, IX-2 treated with Art were inhibited in a concentration-dependent manner(P < 0.01). Art could significantly increase the content of cerarnide in LX-2 ( P <0.01), and the content of Hyp was significantly decreased (P <0.05, P <0.01). The expressions of PPAR-γ, p53 and Caspase 3 were increased compared with that of control group(P < 0.01). CONCLUSION: Artesunate could inhibit the proliferation and induce apoptosis of hepatic stellate cells through upregulating ceramide.


Subject(s)
Apoptosis , Artemisinins/pharmacology , Cell Proliferation , Hepatic Stellate Cells/drug effects , Artesunate , Caspase 3/metabolism , Cell Line , Ceramides/metabolism , Humans , Hydroxyproline/metabolism , Liver Cirrhosis , PPAR gamma/metabolism
5.
World J Gastroenterol ; 19(32): 5326-33, 2013 Aug 28.
Article in English | MEDLINE | ID: mdl-23983437

ABSTRACT

AIM: To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills (FFBJRGP) on hepatic fibrosis in vivo and in vitro. METHODS: Hepatic fibrosis was induced by carbon tetrachloride composite factors. Adult Wistar rats were randomly divided into four groups: normal control group; hepatic fibrosis model group; FFBJRGP-treated group at a daily dose of 0.55 g/kg; and colchicine-treated group at a daily dose of 0.1 g/kg. The effects of FFBJRGP on liver function, serum levels of hyaluronic acid (HA), type IV collagen (CIV), type III procollagen (PC III), laminin (LN), histopathology, and expression of transforming growth factor (TGF-ß1) and Smad3 in hepatic fibrosis were evaluated in vivo. The effects of FFBJRGP on survival rate, hydroxyproline content and cell cycle distribution were further detected in vitro. RESULTS: Compared with the hepatic fibrosis model group, rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions. Compared with those of the model group, the activities of alanine aminotransferase (62.0 ± 23.7 U/L) and aspartate aminotransferase (98.8 ± 40.0 U/L) in the FFBJRGP-treated group were decreased (50.02 ± 3.7 U/L and 57.2 ± 30.0 U/L, respectively, P < 0.01). Compared with those in the model group, the levels of PCIII (35.73 ± 17.90 µg/mL), HA (563.82 ± 335.54 ng/mL), LN (89.57 ± 7.59 ng/mL) and CIV (29.20 ± 6.17 ng/mL) were decreased to 30.18 ± 9.41, 456.18 ± 410.83, 85.46 ± 7.51 and 28.02 ± 9.45 ng/mL, respectively. Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-ß1 and Smad3 were down-regulated in vivo. Cell proliferation was inhibited, the level of hydroxyproline was decreased compared with the control group (P < 0.01), and the cell cycle was redistributed when exposed to FFBJRGP in vitro. CONCLUSION: FFBJRGP inhibits hepatic fibrosis in vivo and in vitro, which is probably associated with downregulation of fibrogenic signal transduction of the TGF-ß-Smad pathway.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Hepatocytes/drug effects , Liver Cirrhosis, Experimental/drug therapy , Liver/drug effects , Medicine, Chinese Traditional , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Biomarkers/blood , Carbon Tetrachloride , Cell Cycle/drug effects , Cell Line , Cell Proliferation/drug effects , Collagen Type III/blood , Collagen Type IV/blood , Female , Hepatocytes/metabolism , Hepatocytes/pathology , Humans , Hyaluronic Acid/blood , Hydroxyproline/metabolism , Laminin/blood , Liver/metabolism , Liver/pathology , Liver Cirrhosis, Experimental/chemically induced , Liver Cirrhosis, Experimental/metabolism , Liver Cirrhosis, Experimental/pathology , Male , Procollagen/blood , Rats , Rats, Wistar , Smad3 Protein/metabolism , Tablets , Transforming Growth Factor beta1/metabolism
6.
Zhonghua Gan Zang Bing Za Zhi ; 20(4): 294-9, 2012 Apr.
Article in Chinese | MEDLINE | ID: mdl-22964152

ABSTRACT

OBJECTIVE: To investigate the impact of the Artemisia annua plant-derived drug, artesunate, on proliferation of primary rat hepatic stellate cells (HSCs), and to analyze the underlying molecular mechanisms of its anti-fibrogenic effects involving the inhibition of transforming growth factor-beta 1 (TGF-b1) expression and secretion in liver. METHOD: Isolated, cultured, and activated primary rat HSCs were divided into sixteen groups, including one untreated control group and fifteen artesunate-treated experimental groups with 125, 150, 175, 200 or 225 mumol/L for 24, 48 or 72 hours. The rate of cellular proliferation was measured using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. TGF-b1 mRNA expression was evaluated by reverse transcription-polymerase chain reaction and protein expression was evaluated by Western blotting. Enzyme-linked immunosorbent assay was used to evaluate secreted levels of TGF-b1 protein. RESULTS: Artesunate significantly inhibited proliferation of cultured HSCs in a dose- and time-dependent manner (all, P less than 0.01). After 24 hours of exposure, the inhibition ratios of the various artesunate concentrations were: 6.06%+/-1.44% (125 mumol/L), 21.47%+/-5.57% (150 mumol/L), 42.00%+/-7.36% (175 mumol/L), 67.12%+/-4.55% (200 mumol/L), and 79.83%+/-3.67% (225 mumol/L). Artesunate significantly inhibited the TGF-b1 mRNA expression in HSCs, and the higher the drug concentration, the higher the degree of inhibition (all, P less than 0.01). In addition, artesunate significantly inhibited the expression of intracellular and secreted TGF-b1 protein (all, P less than 0.01). In response to artesunate (mumol/L concentrations), the TGF-b1 levels were (164.24+/-6.88) pg/ml (0µmol/L), (102.68+/-4.45) pg/ml (150µmol/L), (86.54+/-5.56) pg/ml (175µmol/L), and (56.55+/-5.66) pg/ml (200µmol/L). CONCLUSION: Artesunate exerts anti-fibrogenic effects on HSCs in vitro, possibly by reducing the expression, translation and secretion of TGF-b1.


Subject(s)
Artemisinins/pharmacology , Hepatic Stellate Cells/drug effects , Hepatic Stellate Cells/metabolism , Transforming Growth Factor beta1/metabolism , Animals , Artesunate , Cells, Cultured , Rats , Rats, Wistar
7.
Chinese Journal of Cardiology ; (12): 115-119, 2012.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-275092

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the relationship between 25-hydroxy vitamin D [25(OH)D] and carotid artery intimal medial thickness (IMT) in type 2 diabetic (T2DM) patients.</p><p><b>METHODS</b>Serum 25(OH)D and carotid IMT were measured in 300 T2DM patients. Patients were divided into four quartile groups according to the serum 25(OH)D levels (Q1: < 26.17 nmol/L, 74 cases; Q2: 26.17 - 32.75 nmol/L, 76 cases; Q3: 32.75 - 42.93 nmol/L, 78 cases; Q4 > 42.93 nmol/L, 72 cases).</p><p><b>RESULTS</b>Carotid IMT, carotid artery plaque prevalence, duration of diabetes, HbA1c, CRP and PTH were significantly higher in subjects with low 25(OH)D compared subjects with high 25(OH)D (P < 0.05). Carotid artery IMT in Q1 and Q2 groups were significantly higher than that in Q4 group (1.03 ± 0.21 vs. 0.90 ± 0.20, 1.01 ± 0.26 vs. 0.90 ± 0.20, P < 0.05), was similar among Q1 and Q2 and Q3 groups. Prevalence of carotid atherosclerotic plaque in Q1 group (50.0%) was also significantly higher than in Q3 (29.5%, P < 0.05) and Q4 (16.7%, P < 0.05). Similarly, 25(OH)D concentration was significantly lower in patients with carotid plaque compared patients without carotid plaque [(28.31 ± 4.91) nmol/L vs. (36.31 ± 4.31) nmol/L, P < 0.01]. Pearson correlation analysis showed that carotid IMT was positively correlated with age, smoking, BMI, HbA1c, CRP, LDL-C, PTH/25(OH)D ratio (P < 0.05), and was negatively correlated with 25 (OH) D (r = -0.51, P < 0.01). Multivariate regression analysis showed that 25(OH)D concentration was an independent predictor of carotid IMT in this cohort (β = -0.39, P < 0.01).</p><p><b>CONCLUSION</b>Serum 25(OH)D concentration is negatively correlated with carotid IMT and low 25 (OH) D level is a risk factor for preclinical atherosclerosis in T2DM patients.</p>


Subject(s)
Aged , Female , Humans , Male , Middle Aged , Carotid Intima-Media Thickness , Diabetes Mellitus, Type 2 , Blood , Diagnostic Imaging , Pathology , Vitamin D , Blood
8.
Journal of Experimental Hematology ; (6): 1205-1211, 2012.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-278405

ABSTRACT

This study was purposed to investigate the intercellular cell adhesion molecule-1 (ICAM-1) gene K469E (A/G) (rs5498) and K56M (A/T) (rs5491) single nucleotide polymorphisms (SNP) and soluble ICAM-1 (sICAM-1) levels in plasma in three Chinese populations of Yugur, Tibetan and Han nationalities, to analyze comparatively the genotypes and allele frequencies distribution in different ethnic groups, and to explore the effects of ICAM-1 K469E and K56M polymorphism and sICAM-1 levels in plasma. EDTA-anticoagulant venous blood from Yugur(327 cases), Tibetan (400 cases) and Han (126 cases) people was collected, the DNA was extracted by using whole blood genomic DNA extraction kit, DNA SNP were analyzed by PCR-RFLP, genotype was judged by gel scan imaging system after agarose gel electrophoresis, the gene sequence was determined and the distribution of ICAM-1 genotypes and allele frequencies were compared among different ethnic groups, besides, the group representativeness was tested via the Hardy-Weinberg genetic equilibrium. Finally, the human sICAM-1 plasma levels were detected by using human ICAM-1 ELISA kit. The results showed that DNA sequencing result was consistent with PCR-RFLP analysis. In Yugur, Tibetan and Han nationalities, the KK, KE and EE three genotypes at ICAM-1 K469E gene locus were detected, the genotype distribution was not statistically significantly different, while the K, E allele frequency distribution was statistically significantly different (P < 0.05). Both of genotype and allele frequency distribution between Yugur, Tibetan and Han nationalities were statistically significantly different (P < 0.05). In K56M site only KK, KM two genotypes were detected, but the MM genotype was not detected in the three ethnic groups; the difference of two genotypes and K, M allele frequencies between Yugur and Han population was statistically significantly different (P < 0.05). Among three ethnic groups, the sex ratio and age distribution of K469E, K56M genotypes and allele frequencies of ICAM-1 gene were not significantly different, and distribution was in accordance with Hardy-Weinberg genetic equilibrium (P > 0.05). The plasma sICAM-1 level at ICAM-1 K469E allele locus in K individuals [(253 ± 122), (185 ± 97) µg/L] was higher than that at non-K allele [(145 ± 110) µg/L, P < 0.01]; the plasma sICAM-1 level of ICAM-1 K56M sites with KK genotype [(253 ± 122) µg/L] was higher than that of the KM genotypes [(168 ± 103) µg/L, P < 0.01]. In Yugur and Tibetan groups, the plasma sICAM-1 levels [(224 ± 80), (214 ± 111) µg/L] were higher than that in the Han group [(175 ± 125)µg/L, P < 0.05]. Pairwise comparison indicated that the plasma sICAM-1 levels between Yugur and Han group were statistically significantly different (P < 0.01), that was significantly different between Tibetan and Han group (P < 0.05). It is concluded that in Yugur, Tibetan and Han population, the genotypes and gene frequencies of two amino acid sites K469E and K56M in ICAM-1 were KK/KE-type, KK-type and K allele, moreover, the ratio of them in Yugur and Tibetan group was higher than that in Han, while there is not significant difference in sex ratio and age distribution, therefore, ICAM-1 genotype and allele frequency distribution in this study had ethnic representativeness. ICAM-1 gene K469E and K56M polymorphisms were likely to affect the plasma sICAM-1 expression level. K469E gene K allele may be a genetic risk factor, while K56M gene M allele a may be genetic protective factor for some diseases.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , Ethnicity , Genetics , Gene Frequency , Genotype , Intercellular Adhesion Molecule-1 , Blood , Genetics , Plasma , Metabolism , Polymorphism, Genetic
9.
Journal of Experimental Hematology ; (6): 1200-1204, 2012.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-278406

ABSTRACT

The aim of this study was to investigate the similarities and differences of A1381T (rs216311) and -1793G/C (rs7966230) single nucleotide polymorphisms (SNP) in Chinese Yugur, Tibetan, and Han nationalities and their influence on plasma vWF concentration in order to explore the sensitivity of these 3 nationalities to vWF-related diseases. Peripheral venous blood was obtained from 322 Yugur, 399 Tibetan, and 120 Han healthy people. The DNA were then extracted. vWF gene A1381T and -1793G/C polymorphisms were analyzed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequenced when it was necessary. The vWF:Ag level in plasma was determined by ELISA. The results showed that the genotype distribution of vWF gene at both A1381T and -1793G/C loci in Yugur, Tibetan and Han nationalities was different with statistically significance (P < 0.05). GG genotype of A1381T locus accounted for 69.9% in Yugur nationality, which was much higher than 56.6% and 53.3% in Tibetan and Han nationalities respectively(P < 0.01); AA genotype of A1381T locus expressed a low level of vWF in plasma. For the -1793G/C locus, the proportion of CG genotype in Yugur was much higher than that in Han, CC genotype expressed a high level of vWF in plasma. The plasma vWF levels with different nationalities and the polymorphism of vWF gene were significantly different. It is concluded that the polymorphisms of vWF gene at both A1381T and -1793G/C loci in Yugur, Tibetan and Han are significantly different; the polymorphism of vWF gene influences the plasma vWF level; the plasma vWF levels in Yugur and Tibetan are significantly higher than that in Han, which may be associated with the living environment and habits.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , China , Ethnicity , Genetics , Gene Frequency , Genotype , Plasma , Chemistry , Polymorphism, Genetic , von Willebrand Factor , Genetics , Metabolism
10.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-244959

ABSTRACT

Different cytokines are needed in the course of culturing cells to do adoptive immunotherapy. This study was aimed to investigate the differentiation directions of lymphocytes and related gene expression characteristics after combined stimulation of lymphocytes by different cytokines or EBV antigen peptide combined with cytokines. The experiment was divided into 4 groups. The levels of total T lymphocytes (CD3(+)), T helper lymphocytes (CD3(+)CD4(+)), cytotoxic T-lymphocyte (CD3(+)CD8(+)), memory T cells (CD3(+)CD8(+)CD45RO(+)), naive T cells (CD3(+)CD8(+)CD45RA(+)), Th2 cells (CD3(+)CD30(+)), B cells (CD19(+)), NK cells (CD56(+)), naive T regulatory cells (CD4(+)CD25(+)), precise T regulatory cells (CD4(+)CD25(+)FOXP3(+)) were detected by flow cytometry. The expression levels of house-keeping gene (mad1, pten), T helper cells transcriptional regulatory gene t-bet (Th1), gata3 (Th2), cytokine IFN-γ(Th1), IL-4(Th2) were detected by using RT-PCR. The results showed that CTL in EBV polypeptide group were dominant cells with certain clinical effects. Comparison of result of EBV polypeptide group with other 3 different cytokine stimulating groups demonstrated that EBV antigen peptide had much more effects on stimulating CTL generation. The expression of IFN-γ gene was significantly increased; the T helper differentiation-related gene t-bet, gata3 also increased evidently, while expression change of house-keeping gene mad1 and pten were not evident. Addition of different cytokines and antigen peptides in culture may be much more effective on stimulating CTL generation. It is concluded that specific CTL can be obtained by using the lymphocytes co-cultured with EBV and cytokines, and the different cytokines play different roles in cell differentiation.


Subject(s)
Humans , Cells, Cultured , Cytokines , Allergy and Immunology , Metabolism , Epstein-Barr Virus Nuclear Antigens , Genetics , Flow Cytometry , Immunotherapy, Adoptive , Lymphocyte Count , Lymphoma, Extranodal NK-T-Cell , Genetics , Allergy and Immunology , T-Lymphocytes, Cytotoxic , Allergy and Immunology
11.
World J Gastroenterol ; 16(12): 1458-64, 2010 Mar 28.
Article in English | MEDLINE | ID: mdl-20333785

ABSTRACT

AIM: To explore the anti-fibrotic effect of Haobie Yangyin Ruanjian decoction (HYRD) on CCl(4)-induced hepatic fibrosis in rats and its modulation on the transforming growth factor (TGF) beta-Smad signaling pathway. METHODS: Fifty-six healthy Wistar rats were randomly divided into five groups: normal control group (n = 6), CCl(4)-induced hepatic fibrosis group (n = 14) and three treatment groups (the treated rats received HYRD via oral administration at daily dosages of 8.2, 2.5 and 0.82 g/kg, respectively) of HYRD (n = 12, respectively). Experimental hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride solution (CCl(4) dissolved in peanut oil, 4:6, V/V) with 0.5 mL/100 g body weight for the first time, and then 0.3 mL/100 g body weight twice a week for 8 wk. In the former 2 wk, rats were raised by feedstuff I (80% corn meal, 20% lard, 0.5% cholesterol). After 2 wk, they were raised by feedstuff II (corn meal and 0.5% cholesterol). Except for the control group, 30% alcohol solution was given orally to each rat every other day from the beginning, 1 mL for each rat. Liver function parameters and hepatic hydroxyproline content were detected by chromatometry. Serum levels of hyaluronic acid (HA), type IV collagen (CIV), type III precollagen (PCIII) and laminin (LN) were assayed with radioimmunoassay. Deposition of collagen was observed with hematoxylin-eosin staining and collagen staining. Gene expression of TGFbeta1 and Smad3 were detected with real-time reverse transcriptase-polymerase chain reaction and Western blotting, respectively. RESULTS: The serum levels of alanine transaminase and aspartate transaminase were increased in the model group compared with the control group (P < 0.01), and they were decreased in the three treatment groups compared with the model group. The serum levels of total protein and albumin were decreased in the model group and increased in the three treatment groups. The hepatic hydroxyproline content and serum levels of PCIII, HA, LN and CIV were markedly increased in the model group compared with the control group, and decreased in the treatment groups. The gene expression of TGFbeta1 and Smad3 was enhanced in the model group compared with the control group, and HYRD could down regulate their expression. CONCLUSION: HYRD can inhibit hepatic fibrosis induced by CCl(4) in rats, which is probably associated with its down-regulation on fibrogenic signal transduction of TGFbeta-Smad pathway.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Liver Cirrhosis/prevention & control , Liver/drug effects , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Biomarkers/blood , Blotting, Western , Carbon Tetrachloride , Collagen Type III/blood , Collagen Type IV/blood , Disease Models, Animal , Female , Hyaluronic Acid/blood , Hydroxyproline/metabolism , Laminin/blood , Liver/metabolism , Liver/pathology , Liver Cirrhosis/chemically induced , Liver Cirrhosis/genetics , Liver Cirrhosis/metabolism , Liver Cirrhosis/pathology , Male , RNA, Messenger/metabolism , Radioimmunoassay , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effects , Smad3 Protein/genetics , Transforming Growth Factor beta1/genetics
12.
Zhonghua Gan Zang Bing Za Zhi ; 18(2): 113-8, 2010 Feb.
Article in Chinese | MEDLINE | ID: mdl-20196950

ABSTRACT

OBJECTIVE: To explore therapeutic effect of Haobieyangyinruanjianfang (HBYYRJ) on mouse liver fibrosis by schistosomiasis. METHODS: Mice except for normal control were infected with Japanese schistosome cercarias, after 12 weeks, infected mice were divided into 7 groups: low HBYYRJ group, middle HBYYRJ group, high HBYYRJ group, Fufangbiejiaruangan tablet (FFBJRG) group, colchicine group, 3 months infection group and 6 months infection group. Hepatic fibrosis was found in 3 months infection group. Liver hydroxyproline (Hyp) was determined, matrix metalloproteinase-2 and 9 (MMP-2, MMP-9) were detected with gelatin zymography, serum hyaluronic acid (HA) and precollagen III (PC-III) were detected using RIA. RESULTS: HBYYRJ obviously reduced hepatic fibrosis (probability value less than 0.01). Collagen and HA in 3 months infection group and 6 months infection group were higher than that in normal group (probability value less than 0.01), collagen in high and middle HBYYRJ groups and HA in middle and low HBYYRJ groups were lower than that in 6 months infection group (P less than 0.01, probability value less than 0.05). The expression of MMP-9 and MMP-2 in 3 months infection group and 6 months infection group was higher than that in normal group (probability value less than 0.01), The expression of MMP-9 in three HBYYRJ groups and the expression of MMP-2 in high HBYYRJ group were lower than that in 6 months infection group (probability value less than 0.05). CONCLUSION: HBYYRJ can reduce liver fibrosis caused by schistosomiasis.


Subject(s)
Drugs, Chinese Herbal/therapeutic use , Liver Cirrhosis, Experimental/drug therapy , Liver/drug effects , Materia Medica/therapeutic use , Schistosomiasis japonica/complications , Animals , Collagen Type III/blood , Disease Models, Animal , Drugs, Chinese Herbal/isolation & purification , Drugs, Chinese Herbal/pharmacology , Female , Hyaluronic Acid/blood , Hydroxyproline/metabolism , Liver/metabolism , Liver/pathology , Liver Cirrhosis, Experimental/etiology , Liver Cirrhosis, Experimental/metabolism , Liver Cirrhosis, Experimental/pathology , Male , Materia Medica/isolation & purification , Materia Medica/pharmacology , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mice , Schistosoma japonicum , Severity of Illness Index , Sex Factors , Treatment Outcome
13.
Chinese Medical Journal ; (24): 486-495, 2009.
Article in English | WPRIM (Western Pacific) | ID: wpr-311835

ABSTRACT

<p><b>BACKGROUND</b>The presence of autoantibodies against multiple epidermal proteins is an important feature in paraneoplastic pemphigus (PNP). Circulating anti-desmoglein 3 autoantibody, the major pathogenic autoantibody in pemphigus vulgaris (PV), has been proved pathogenic in PNP. Because of many clinical differences between PNP and PV, we speculate about the involvement of other autoantibodies in the pathogenesis of PNP. Envoplakin (EPL) and periplakin (PPL) are recognized by most PNP sera. Their linker subdomains are highly homologous and necessary for the association of intermediate filaments.</p><p><b>METHODS</b>We characterized the autoantibodies against the linker subdomains of EPL and PPL in PNP patients' sera and their associated tumors by enzyme-linked immunosorbent assay (ELISA) and immunofluorence. We also applied the purified autoantibodies against EPL and PPL from PNP sera to cultured human epidermal keratinocytes (HEK), to evaluate the changes of cell-cell adhesion.</p><p><b>RESULTS</b>Autoantibodies against EPL and PPL were detected in most PNP patients by ELISA, and the decrease of these autoantibodies after removal of the tumors was roughly comparable to the improvement of clinical symptoms. Cultured tumor cells from PNP patients secreted these autoantibodies. Specific immunoglobulin receptors for EPL and PPL were found on B lymphocytes in tumors from PNP. Furthermore, purified anti-EPL and anti-PPL autoantibodies from PNP sera were capable of dissociating cultured human epidermal keratinocytes.</p><p><b>CONCLUSION</b>Autoantibodies against EPL and PPL may also be pathogenic in PNP.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Autoantibodies , Allergy and Immunology , Pharmacology , Cell Adhesion , Cells, Cultured , Desmoglein 3 , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Epidermis , Cell Biology , Keratinocytes , Cell Biology , Membrane Proteins , Allergy and Immunology , Paraneoplastic Syndromes , Allergy and Immunology , Metabolism , Pemphigus , Allergy and Immunology , Metabolism , Plakins , Allergy and Immunology , Protein Precursors , Allergy and Immunology
14.
Chinese Journal of Cardiology ; (12): 243-248, 2008.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-243805

ABSTRACT

<p><b>OBJECTIVE</b>We investigated the in vivo effects of recombinant adenovirus-associated virus type-2 (AAV-2) mediated interleukin-10 (IL-10) gene transfer on the expression of matrix metalloproteinase (MMP)-2, 9, tissue inhibitor of metalloproteinase (TIMP)-1, collagen type I and type III in a rat acute myocardial infarction model.</p><p><b>METHOD</b>Male Sprague-Dawley (SD) rats were randomly divided into three groups (each n = 6): sham operation group, MI/AAV2 group, and MI/AAV2-IL-10 group (10(10) vg/ml x 0.1 ml injection at peri-infarct regions immediately post MI). Five days later, the expressions of MMP-2 and MMP-9 were measured by RT-PCR, Western blot and zymography. The expression of TIMP-1 was measured by RT-PCR and Western blot. Collagen type I and type III were assessed by RT-PCR and immunohistochemical stain.</p><p><b>RESULTS</b>The myocardial expressions of MMP-2, MMP-9 and collagen contents in MI/AAV2 group were significantly increased than those in sham operation group. Myocardial expressions of MMP-2, MMP-9 were significantly decreased and the expression of TIMP-1 significantly increased in the MI/AAV2-IL-10 group than those in MI/AAV2 group. Moreover, the expressions of collagen type I, collagen type III and the ratio of I/III collagen in border zones of infarcted myocardium were decreased by 47.6% (P < 0.01), 23.6% (P < 0.05), and 17.9% (P < 0.05) respectively, while the expression of TIMP-1 increased by 73.1%(P < 0.05) in MI/AAV2-IL-10 group compared to MI/AAV2 group.</p><p><b>CONCLUSION</b>In vivo myocardial IL-10 transfer reduced myocardial MMP and collagen expression and increasing the TIMP expression.</p>


Subject(s)
Animals , Male , Rats , Extracellular Matrix , Metabolism , Gene Expression , Genetic Therapy , Interleukin-10 , Genetics , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Myocardial Infarction , Genetics , Metabolism , Myocardium , Metabolism , Rats, Sprague-Dawley , Tissue Inhibitor of Metalloproteinase-1 , Metabolism , Transfection , Ventricular Remodeling
16.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-247291

ABSTRACT

<p><b>OBJECTIVE</b>To conduct a molecular epidemiological survey on the mitochondrial DNA C1494T mutation in non-syndromic hearing loss patients in Chinese population.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to screen the mitochondrial DNA 12S rRNA C1494T mutation in 20 patients with aminoglycoside antibiotic induced hearing loss, 136 sporadic non-syndromic hearing loss patients and 50 probands of pedigrees with non-syndromic hearing loss.</p><p><b>RESULTS</b>The C1494T mutation did not appear in all cases except for the positive control.</p><p><b>CONCLUSION</b>Incidence of mitochondrial DNA C1494T mutation is much lower than that of mitochondrial DNA A1555G mutation in non-syndromic hearing loss of Chinese population. Mitochondrial DNA C1494T mutation may be a rare variation in non-syndromic hearing loss and is not the main cause of aminoglycoside antibiotic induced-deafness.</p>


Subject(s)
Adolescent , Child , Female , Humans , Male , Aminoglycosides , Anti-Bacterial Agents , Asian People , Genetics , China , DNA, Mitochondrial , Genetics , Hearing Loss , Ethnology , Genetics , Point Mutation , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , RNA, Ribosomal , Genetics
17.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-253081

ABSTRACT

<p><b>AIM</b>To explore the time-dependent changes of endogenous hydrogen sulfide system at the early stage of pulmonary hypertension induced by high pulmonary flow in rats.</p><p><b>METHODS</b>Eighty male SD rats, whose weight ranged 140 - 160 g, were randomly divided into control group (n = 40) and shunt group (n = 40). Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. After 1 d, 3 d, 1 week, 4 week and 8 weeks of experiment, systolic pulmonary artery pressure (SPAP) of each rat, the H2S of rat lung tissue and CSEmRNA of rat lung tissue were evaluated, respectively.</p><p><b>RESULTS</b>SPAP increased significantly as compared with those in control group in 1 week and 8 weeks of experiment. In contrast to control group, the H2S of rat lung tissue increased significantly on 3 d and in 4 weeks, respectively. Meanwhile, in contrast to control group, relative amount of CSE mRNA of lung tissues elevated significantly on 3 d and in 4 weeks, respectively. Moreover, SPAP and the H2S of rat lung tissue, the CSE mRNA of rat lung tissue correlated negatively in 1 week, 4 weeks and 8 weeks of experiment.</p><p><b>CONCLUSION</b>Animal model of rats with high pulmonary blood flow exhibited pulmonary hypertension. Lung tissue H2S and CSE mRNA of rats exhibited double peaks within 8 weeks. These results revealed that endogenous H2S system might be relevant with the development of pulmonary hypertension induced by high pulmonary blood flow, and probably, it played a protective role in the regulation of pulmonary hypertension, especially, at its early stage.</p>


Subject(s)
Animals , Male , Rats , Hydrogen Sulfide , Metabolism , Hypertension, Pulmonary , Metabolism , Lung , Metabolism , Pulmonary Artery , Metabolism , Rats, Sprague-Dawley
18.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-313719

ABSTRACT

<p><b>OBJECTIVE</b>To suppress the expression of CCR5 and CXCR4, the co-receptors for human immunodeficiency virus type 1 ( HIV-1), and thus inhibit HIV-1 from entering cells.</p><p><b>METHODS</b>DNA fragments encoding either CCR5 or CXCR4 were amplified from healthy human peripheral blood mononuclear cells (PBMCs) by reverse transcript polymerase chain reaction (RT-PCR) and sequencing was performed. Correct fragments were inserted into Shuttle plasmid inversely, which was recombined with backbone plasmid containing homologous adenoviral genome in E. coli BJ5183. The recombinant plasmids were transfected into 293 cells in which they were packaged and amplified. Recombinant adenoviruses containing antisense RNA of CCR5 or CXCR4 were obtained and identified by RT-PCR, and the titres of them were determined by cytopathic effect (CPE) method. The U937 and MT4 cells were infected by recombinant adenoviruses containing antisense RNA of CCR5 (multiplicity of infection, MOI = 100) and CXCR4 (MOI = 200), respectively. The expression of co-receptors on infected cell was measured by fluorescence activated cell sorter at 24, 48, 72 hours and 10 days after infection. In addition, the chemotactic activity and proliferation of infected cells were detected with Boyden chamber and 3H incorporation respectively.</p><p><b>RESULTS</b>We constructed the recombinant plasmids and obtained the recombinant adenoviruses which contained antisense RNA of CCR5 or CXCR4 and were designated as pAd-antiR5 and pAd-antiX4 respectively. The titers of recombinant adenoviruses pAd-antiR5 and pAd-antiX4 were 5 x 10" PFU/ml and 7 x 10(10) PFU/ml, respectively. The expression rate of CCR5 on U937 cells decreased from 82. 10% (blank control) to 1.12% (Ad-antiR5 infected) , and that of CXCR4 on MT4 cells decreased from 42% (blank control) to 1.03% (Ad-antiX4 infected) 24 hours later. The expression rates of CCR5 on Ad-antiR5 infected U937 cells were 1.02% , 1.26% , 1.23% at 48 hours, 72 hours, and 10 days later, respectively. The expression rates of CXCR4 on Ad-antiX4 infected MT4 cells were 1.13%, 1.17%, 1.22% at 48 hours, 72 hours, and 10 days later, respectively. Moreover, the recombinant adenovirus had no effects on chemotactic activity and proliferation of the cells.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing antisense CCR5 or CXCR4 can remarkably decrease the expression of co-receptors for HIV-1 on U937 or MT4 cells without affecting their chemotactic activities and proliferative abilities.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Cell Line, Transformed , Cell Proliferation , Chemotaxis , Down-Regulation , Genetic Vectors , RNA, Antisense , Genetics , RNA, Messenger , Genetics , Receptors, CCR5 , Genetics , Receptors, CXCR4 , Genetics , Transfection , U937 Cells
19.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-262763

ABSTRACT

<p><b>OBJECTIVE</b>Febrile seizure (FS) is one of the most common seizure types in children. Our previous studies have demonstrated that both gamma-aminobutyric acid B receptor (GABABR) and hydrogen sulfide (H2S) are involved in the pathogenesis of FS. This study was designed to explore the effect of GABABR on H2S/cystathionine-beta-synthase (CBS) system in recurrent FS.</p><p><b>METHODS</b>Sixty-four Sprague-Dawley rats aged 21 days were randomly assigned into four groups: Control (37 degrees C water bath exposure), FS, FS+baclofen (GABABR excitomotor), and FS+phaclofen (GABABR inhibitor) groups (n=16 each). FS was induced by warm water bath exposure (45.2 degrees C, once every 2 days, 10 times in total. The plasma level of H2S was detected by the spectrophotometer. The expression of CBS mRNA was examined by in situ hybridization. The expressions of CBS protein was observed by immunohistochemistry.</p><p><b>RESULTS</b>The plasma level of H2S increased in the FS+baclofen group (427.45 +/- 15.91 micromol/L) but decreased in the FS+phaclofen group (189.72 +/- 21.53 micromol/L) compared with that in the FS group (362.14 +/- 19.71 micromol/L). The expressions of CBS mRNA and protein were up-regulated in the FS+baclofen group but were down-regulated in the FS+phaclofen group compared with those in the FS group.</p><p><b>CONCLUSIONS</b>GABABR modulated the expression of H2S/CBS system in recurrent FS.</p>


Subject(s)
Animals , Rats , Baclofen , Pharmacology , Cystathionine beta-Synthase , Genetics , Physiology , Hydrogen Sulfide , Blood , Metabolism , Rats, Sprague-Dawley , Receptors, GABA-B , Physiology , Recurrence , Seizures, Febrile , Metabolism
20.
Chinese Journal of Pediatrics ; (12): 527-530, 2006.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-278665

ABSTRACT

<p><b>OBJECTIVE</b>Febrile seizure (FS) is closely related to an altered transmission of gamma-aminobutyric acid (GABA). GABA exerts its effects through ionotropic receptors (GABA(AR) and GABA(CR)) and metabotropic receptors (GABA(BR)). GABA(BRs) are located at pre- and postsynaptic sites. Stimulation of postsynaptic receptors generates long-lasting inhibitory postsynaptic potentials (IPSPs) that are important for the fine-tuning of inhibitory neurotransmission and caused by an increase in K(+) conductance. At presynaptic sites, GABA(BRs) mediate a suppression on the release of neurotransmitters such as of GABA or glutamate by inhibiting voltage-sensitive Ca(2+) channels. The present study aimed to explore the long-term changes of GABA(B) receptor subunits in immature rats after recurrent febrile seizures.</p><p><b>METHODS</b>Rats were randomly divided into control group and hyperthermia treatment group. The control rats (n = 64) were put into 37 degrees C water for 5 minutes. Rats with hyperthermia treatment were put into 44.8 degrees C water for 5 minutes. If a rat in hyperthermia treatment group showed seizure within 5 min, the rat was taken out of the water as soon as the seizure occurred. Water-immersion was carried out 10 times, once every 2 days. Rats showing 10 seizures (FS(10), n = 64) were studied. Rats exposed to hyperthermia for 10 times without seizure were also studied as hyperthermia-only (H, n = 64) group. Rats showing one seizure at the last time of 10 times of hyperthermia treatment were studied as one-seizure group (FS(1), n = 64). The other rats were studied for other research. The changes of GABA(B)R(1) and GABA(B)R(2) co-localization were detected by double fluorescence;the quantitative alteration of GABA(B)R(1) and GABA(B)R(2) were detected by quantitative RT-PCR; the binding of GABA(B)R(2) to GABA(B)R(1) was detected by immunoprecipitation/Western blot.</p><p><b>RESULTS</b>GABA(B)R(1), GABA(B)R(2), and the binding of GABA(B)R(2) to GABA(B)R(1) decreased after the last febrile seizure in FS(10) group, the expression of GABA(B)R(1) returned to normal in later phase while GABA(B)R(2) and the binding of them did not.</p><p><b>CONCLUSION</b>Recurrent FS down-regulated the expression of GABA(B)R subunits in a long term.</p>


Subject(s)
Animals , Rats , Age Factors , Blotting, Western , Disease Models, Animal , Down-Regulation , Fluorescent Antibody Technique , Hippocampus , Metabolism , Immunoprecipitation , RNA, Messenger , Rats, Sprague-Dawley , Receptors, GABA-B , Classification , Genetics , Metabolism , Recurrence , Reverse Transcriptase Polymerase Chain Reaction , Seizures, Febrile , Genetics , Metabolism , Time Factors
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