2.The study of mRNA expression of adiponectin in omental adipose tissue of patients with type 2 diabetes
Hong LIU ; Anping WANG ; Ning XIA
Chinese Journal of Diabetes 2008;16(8):471-474
Objective To study the adiponectin mRNA expression in omental adipose tissue of type 2 diabetes. Methods RT-PCR method was used to examine the adiponectin mRNA expression. Fasting levels of BG,TG,TC,HDL-C,LDL-C of all the subjects were checked and their blood pressure,height,weight,waist circumference,hip circumference were measured to calculate BMI,WHR. Results The adiponectin mRNA expression in omental adipose tissue was decreased in diabetes group versus non-diabetes group (P<0.05),and negatively correlated with WC,TG,FBG,WHR(P<0.05). Conclusions Type 2 diabetic patients show lower expression of adiponectin mRNA in omental adipose tissue than that of non-diabetic control.WC,TG,FBG, WHR and course of disease are correlated with adiponectin mRNA expression and insulin resistance
3.99Tcm-MIBI SPECT for the differential diagnosis of parotid tumors
Chinese Journal of Nuclear Medicine 2011;31(5):310-312
Objective To evaluate the clinical value of 99Tcm-MIBI SPECT for the differential diagnosis of parotid tumors before treatment.Methods 99Tcm-MIBI SPECT imaging of the parotid region was obtained in 32 patients with parotid tumors before surgery.Early and delayed 99Tcm-MIBI imaging were performed in all patients and the diagnosis was confirmed by pathological results after surgery.The ratio of radioactivity between the tumor and opposite side of normal parotid tissue (T/N) was measured.Fisher exact probability test and t test were used for statistical analysis.Results The sensitivity,specificity and accuracy of 99Tcm-MIBI SPECT imaging for the differential diagnosis of parotid tumor were 90.00% (9/10),86.36% (19/22) and 87.50% (28/32),respectively.Among 22 patients who had benign tumors,19(86.36%) showed negative findings,and the other 3 ( 13.64% ) patients showed false positive results.In 10 patients with malignant tumors,1 ( 10.00% ) had false negative findings,and all the other 9 (90.00%)patients showed positive results.The difference between the benign and malignant groups was statistically significant (P =0.00018 ).In the early images,the T/N ratios of benign and malignant parotid tumors were 1.45 ±0.38 and 1.65 ±0.63 (t =20.4,P<0.01),respectively; and in the delayed images,the ratios were 1.43 ± 0.56 and 1.77 ± 0.59 ( t =2.4,P < 0.05 ),respectively.Conclusion 99Tcm-MIBI SPECT imaging might be useful for the differential diagnosis of parotid tumors before surgery.
5.Plum blossom needling for 74 cases of scapulohumeral periarthritis.
Chinese Acupuncture & Moxibustion 2011;31(11):1040-1040
Acupuncture Points
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Adult
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Aged
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Female
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Humans
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Male
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Middle Aged
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Periarthritis
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therapy
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Punctures
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Treatment Outcome
6.Detection and clinical value of novel GATA6 mutations associated with congenital atrial septal defects
Guifen ZHENG ; Hong ZHAO ; Dong WEI ; Ning ZHOU ; Xingyuan LIU
Chinese Journal of Laboratory Medicine 2012;(12):1108-1111
Objective To identify novel mutations in the GATA6 gene associated with congenital atrial septal defects (ASD).Methods This was a case-control study.A cohort of 220 unrelated Han-race patients with congenital ASD and 200 unrelated ethnically matched healthy individuals used as controls,who were admitted to Tongji University Affiliated Tongji Hospital from January,2007 to October,2011,were recruited.The peripheral venous blood samples from the participants were prepared.All the coding exons and their flanking sequences of the GATA6 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination technique.The acquired sequences were aligned with the sequences derived from GenBank by BLAST to identify the sequence variations.The software ClustalW was used to analyze the conservation of the altered amino acids.Results Three novel heterozygous missense GATA6 mutations,c.250G >A (p.A84T),c.649G >C (p.G217R) and c.1270A >C (p.S424R),were identified in 3 of 220 ASD patients,respectively.None of the three mutations was detected in 200 healthy control individuals.A cross-species alignment of GATA6 encoded protein sequences showed that the mutated amino acids were relatively conserved evolutionarily.Conclusion The identification of novel GATA6 mutations associated with ASD contributes to the reveal of the mechanism involved in the pathogenesis of ASD.
7.Effects of combination of glycyrrhizin acid, ligustrazine and puerarin on LPS-induced cytokines expression in macrophage.
Zhao LIU ; Ju-ying ZHONG ; Er-ning GAO ; Hong YANG
China Journal of Chinese Materia Medica 2015;40(20):4068-4074
To study the anti-inflammatory activity of glycyrrhizin acid, ligustrazine and puerarin. In the study, the liquichip-based high-throughput synchronous detection technique for 23 inflammatory factors, uniform design, comprehensive weight method were adopted to study the effect of different combined administration of glycyrrhizin acid, ligustrazine and puerarin in inhibiting the expression of lipopolysaccharide (LPS)-induced RAW264. 7 cells and multiple inflammatory cytokines. In the study, the uniform design table U₉ (9³) was adopted to design doses of glycyrrhizin acid, ligustrazine and puerarin. The liquichip technique was used to detect the effect of different combined administration of glycyrrhizin acid, ligustrazine and puerarin on the 23 cytokines expressed in LPS-induced mouse macrophage RAW264. 7 inflammation model. The traditional Chinese medicine component optimization software and the improved least angle regression algorithm were used to analyze the dose-effect relationship among the three components and the cytokine inhibition rate and produce the regression equation. The comprehensive weight method was applied to get the optimal dose ratio of glycyrrhizic acid, ligustrazine and puerarin with highest efficacy of 25:2:13 and verify the optimal dose ratio. The verification results were consistent with the prediction trend, indicating the accuracy of the mathematical model for predicting the experiment. The experimental results showed the multi-target and multi-level efficacies of glycyrrhizic acid, ligustrazine and puerarin and the high anti-inflammatory activity of their combined administration, which provides powerful basis for subsequent drug development.
Animals
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Anti-Inflammatory Agents
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pharmacology
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Cytokines
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Glycyrrhizic Acid
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pharmacology
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Isoflavones
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pharmacology
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Lipopolysaccharides
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immunology
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Macrophages
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drug effects
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immunology
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Mice
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NF-kappa B
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genetics
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immunology
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Pyrazines
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pharmacology
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RAW 264.7 Cells
8.Effects of glycyrrhizin acid and licorice flavonoids on LPS-induced cytokines expression in macrophage.
Zhao LIU ; Ju-Ying ZHONG ; Er-Ning GAO ; Hong YANG
China Journal of Chinese Materia Medica 2014;39(19):3841-3845
Glycyrrhizin acid and licorice flavonoids are the component of Glycyrrhiza uralensis Fisch root that has been used for various medicinal purposes in traditional oriental medicine for thousands of years. Macrophages as a principal component of immune system play an important role in the initiation, modulation and final activation of immune response against pathogens. In the present study, glycyrrhizin acid and licorice flavonoids was investigated the anti-inflammatory effect on lipopolysaccharide (LPS)-induced macrophage cell line of RAW264.7. Well-grown RAW264.7 cells were collected and randomly divided into the blank control group, the LPS(1 mg x L(-1)) group, the dexamethasone (5 mg x L(-1)) with LPS group, the glycyrrhizin acid (400, 80, 16 mg x L(-1)) with LPS group and the licorice flavonoids (200, 40, 8 mg x L(-1)) with LPS group. RAW264.7 cells were cultured in 24-well plates, pre-incubated for 4 h with different concentrations of dexamethasone, glycyrrhizin acid, or licorice flavonoids. Then cells were stimulated for 20 h with LPS. The supernatant of culture medium was collected from each well and determinated the concentrations of cytokines by means of BioPlex mouse cytokines assay. Compared with the control group, the LPS group could significantly induced relatively high levels of granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor( GM-CSF), macrophage inflammatory protein-1 alpha (MIP-1α), macrophage inflammatory protein-1 beta (MIP-1β), regulated upon activation normal T cell expressed and secreted factor (RANTES), tumor necrosis factor alpha ( TNF-α), monocyte chemotactic protein 1 (MCP-1), chemokine (C-X-C motif) ligand 1 (KC), eotaxin, interleukin(IL)-1α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12 (p40), IL-12 (p70), IL-13, and IL-17 secretion (P < 0.05). The glycyrrhizin acid significantly inhibited IL-1β, IL-3, IL-5, IL-6, IL-10, IL-12 (p40), IL-12 (p70), IL-13, Eotaxin and TNF-α secreted by LPS-stimulated RAW264.7 cells (P < 0.05). The expression levels of IL-6 and Eotaxin were observably decreased in the licorice flavonoids with LPS group (P < 0.05). The data presented here suggested that the glycyrrhizin acid and licorice flavonoids modulate various cytokines secreted by macrophages and were important anti-inflammatory constituent of Licorice.
Animals
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Cell Line
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Cytokines
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genetics
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immunology
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Flavonoids
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pharmacology
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Glycyrrhiza
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chemistry
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Glycyrrhizic Acid
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pharmacology
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Lipopolysaccharides
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immunology
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Macrophages
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drug effects
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immunology
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Mice
9.Protective effect of acetylated epigallocatechin gallate on melanocytes from oxidative stress-induced damage
Weixuan NING ; Suiquan WANG ; Weisong HONG ; Dongyin LIU ; Ai'e XU ;
Chinese Journal of Dermatology 2015;48(7):480-484
Objective To investigate the protective effect of acetylated epigallocatechin gallate (AcEGCG) against H2O2-induced oxidative damage to human epidermal melanocytes,and to explore its possible mechanism.Methods Human epidermal melanocytes were isolated and cultured in vitro.Some melanocytes were classified into a H2O2 group induced by H2O2 only,EGCG groups and AcEGCG groups induced by H2O2 after pretreatment with different concentrations of EGCG and AcEGCG,respectively.Three concentrations (10,20 and 40 μmol/L) of EGCG or AcEGCG were used to treat melanocytes for 1 hour in MTS assay and lactate dehydrogenase (LDH) leakage assay and for 2 hours in Western blot assay,while only one concentration (40 μmol/L) was used to treat melanocytes for 0.5,1,2 and 4 hours respectively in flow cytometry assay.Some melanocytes treated with only culture medium and 0.1% dimethyl sulphoxide (DMSO) served as the control group.After additional culture,MTS assay was performed to determine cell survival rate,flow cytometry to detect the level of reactive oxygen species (ROS) in melanocytes,Western blot to measure the expressions of caspase-9 and caspase-3 proteins.Lactate dehydrogenase (LDH) kit was used to detect the leakage of LDH to culture medium.Statistical analysis was carried out by using one-way analysis of variance for comparisons of multiple group means followed by Student-Newman-Keuls-q (SNK-q) test for multiple comparisons.Results Compared with the control group,the H2O2 group showed significantly decreased cell survival rate (22.99% ± 0.53%,P < 0.01),but increased LDH leakage level (36.58% ± 0.73%,P < 0.01),intracellular ROS level (19.08 ± 0.57,P < 0.01),as well as caspase-9 (2.65 ± 0.079,P < 0.01) and caspase-3 (2.36 ± 0.057,P < 0.01) expressions.In comparison with the H2O2 group,the cell survival rate was significantly higher in the 10-,20-and 40-μmol/L AcEGCG groups (79.50% ± 3.62%,86.52% ± 5.13%,97.81% ± 5.21%,respectively,all P< 0.01) and EGCG groups (43.19% ± 1.68%,63.34% ± 3.60%,70.82% ± 2.1%,respectively,all P < 0.01).However,the 10-,20-and 40-μ mol/L AcEGCG groups and EGCG groups all showed a significant decrease in the expression levels of caspase-9 (AcEGCG groups:1.44 ± 0.067,1.26 ± 0.059 and 1.10 ± 0.072 respectively;EGCG groups:2.31 ± 0.085,2.13 ± 0.091 and 1.35 ± 0.064 respectively,all P < 0.05) and caspase-3 (AcEGCG groups:1.70 ± 0.053,1.57 ± 0.057 and 1.24 t 0.068 respectively,all P< 0.05;EGCG groups:2.09 ± 0.076,1.98 ± 0.093 and 1.79 ± 0.056 respectively,all P < 0.05) compared with the H2O2 group.Similarly,a significant reduction was observed in the leakage level of LDH in these AcEGCG and EGCG groups (all P < 0.01) and in ROS levels in the 40-μmol/L AcEGCG and EGCG groups when compared with the H2O2 group.Conclusions AcEGCG has a stronger protective effect against H2O2-induced oxidative damage to human epidermal melanocytes compared with EGCG,which may be realized through clearance of free radicals,antioxidant effects,and decrease of caspase-9 and caspase-3 expressions.
10.Effects of PP242 on the Expression of Apoptosis Protein Bcl-2 in Human Lens Epithelial Cells
Xue BAI ; Hao FENG ; Liu ZHANG ; Zhibo YANG ; Hong NING
Journal of China Medical University 2016;45(4):324-327
Objective To explore the effects of PP242 on the expression of apoptosis protein Bcl?2 in human lens epithelial cells(HLECs). Meth?ods Immortal HLECs SRA01/04 were cultured and treated with different concentrations of PP242. The cell growth was examined by MTT assay at 24 h and 48 h after PP242 treatment. Real?time quantitative polymerase chain reaction(RT?PCR)and Western blot were adopted to detect the mRNA and protein expression of Bcl?2 respectively. Results The cultured HLECs SRA01/04 were collected and treated with different concentra?tions(100,250,500,750,1 000,1 500 nmol/L)of PP242. After treatment for 24 h and 48 h,the inhibitory rate in each well was determined by MTT assay. The inhibition rate was as below,24 h:7.55%,9.43%,16.98%,22.64%,26.42%,30.19%;48 h:11.11%,23.81%,36.51%,42.86%, 49.21%,63.49%. Compared with the control group,the difference was statistically significant(P<0.05). Compared with the control group(0 nmol/L),the expression of Bcl?2 protein was significantly decreased with the increasing concentrations of PP242(P<0.05),while the expression of Bcl?2 mRNA was inhibited by PP242 based on the results of RT?PCR. Compared with the control group(0 nmol/L),the RT?PCR results of Bcl?2 mRNA were 0.723±0.039,0.517±0.028,0.353±0.052,0.167±0.046,respectively(P<0.05). Conclusion PP242 could inhibit cell proliferation of HLECs in vitro with a concentration and time depended manner. Bcl?2 protein is expressed in HLECs,which could be down regulated by PP242 treatment.