1.The effect of maternal hypothyroidism on microRNA expression profiles in the brain of rat offsprings
Qian XING ; Yun GAO ; Zhongyan SHAN ; Weiping TENG
Chinese Journal of Endocrinology and Metabolism 2013;(1):69-74
Objective To analyse the differential expression of miRNAs in the brain of offsprings of hypothyroid and normal rats,and to explore the molecular mechanism underlying the effect of hypothyroidism on brain development in the offspring.Methods Forty-eight female Wistar rats were assigned to (1) control group (n =24),and (2) hypothyroid group after complete thyroidectomy (n =24).Serum TSH and Total thyroxine (T4) were measured one month after the surgery.Brain samples of fetal or postnatal rats were obtained during four different developmental stages:embryonic days (E) 13,E17,postnatal days (P) 0 and P7.The hippocampus and cortex were separated on P7.MiRNAs were isolated from tissues and two samples were used at each time point studied to reduce the influence of individual differences.The brain samples were detected by Gene Chip miRNA arrays (Affymetrix).Results In the brain tissues of fetal or neonatal rats of maternal hypothyroid rats,two miRNAs (mir-206,-324-5p) on E13,three miRNAs (mir-34c,-204,-194) in cortex on P7,and five miRNAs (mir-146b,-532-5p,-384-5p,-215,-212) in hippocampus on P7 were up-regulated by over 2 folds.Five miRNAs (mir-200b,-200c,-217,-672,-139-5p) on E17,one miRNA (mir-376-3p) on P0,and four miRNA (mir-672,-204,-335,-376-3p) in hippocampus on P7 were decreased by 50% or more.Conclusion The miRNA expression profiles in the rat brain of offspring with maternal hypothyroidism are characterized by miRNA arrays.The identification of a subset of brain expressed miRNAs in the brain may explain the brain development abnormalities resulting from maternal hypothyroidism.
2.Effect of sesamin on pulmonary vascular remodeling in rats with monocrotaline-induced pulmonary hypertension.
Xian-wei LI ; Yun-xing GAO ; Shu LI ; Jie-ren YANG
China Journal of Chinese Materia Medica 2015;40(7):1355-1361
OBJECTIVETo observe the effect of sesamin (Ses) on pulmonary vascular remodeling in rats with monocrotaline ( MCT)-induced pulmonary hypertension (PH).
METHODTotally 48 male Sprague-Dawley (SD) rats were fed adaptively for one week and then divided into the normal control group, the MCT group, the MCT +Ses (50 mg x kg(-1)) group and the MCT + Ses (100 mg x kg(-1)) group, with 12 rats in each group. The PH rat model was induced through the subcutaneous injection with MCT(60 mg x kg(-1)). After the administration for four weeks, efforts were made to measure the right ventricular systolic pressure( RVSP) and mean pulmonary artery pressure (mPAP) through right jugular vein catheterization, and isolate right ventricle( RV) and left ventricle( LV) +septum (S) and measure their length to calculate RV/ ( LV + S) and ratio of RV to tibial length. Pathologic changes in arterioles were observed by HE staining. Masson's trichrome stain was used to demonstrate changes in collagen deposition of arterioles. The alpha-smooth muscle actin (alpha-SMA) expression in pulmonary arteries was measured by immunohistochemisty. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) content in pulmonary arteries were determined by the colorimetric method. The protein expressions of collagen I, NOX2 and NOX4 were analyzed by Real-time PCR and Western blot.
RESULTAfter the administration for 4 weeks, Ses could attenuate RVSP and mPAP induced by MCT, RV/ (LV + S) and ratio of RV to Tibial length, alpha-SMA and collagen I expressions and remodeling of pulmonary vessels and right ventricle. Meanwhile, Ses could obviously inhibit the expressions of NOX2, NOX4 and MDA content and increase T-AOC.
CONCLUSIONSesamin could ameliorate pulmonary vascular remodeling induced by monocrotaline in PH rats. Its mechanism may be related to expressions of NOX2 and NOX4 expression and reduction in oxidative stress injury.
Animals ; Dioxoles ; administration & dosage ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertension, Pulmonary ; drug therapy ; enzymology ; genetics ; physiopathology ; Lignans ; administration & dosage ; Lung ; blood supply ; enzymology ; metabolism ; Male ; Membrane Glycoproteins ; genetics ; metabolism ; Monocrotaline ; adverse effects ; NADPH Oxidase 2 ; NADPH Oxidase 4 ; NADPH Oxidases ; genetics ; metabolism ; Pulmonary Artery ; drug effects ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Vascular Remodeling ; drug effects
3.Puma luciferase reporter gene construction and identification
Xin YANG ; Shi QIU ; Shouzhi GU ; Yun CAI ; Xing GAO ; Zejun LIU
Cancer Research and Clinic 2011;23(1):8-10
Objective To study the mechanism of p55 inducing cell apoptosis, the 180 bp fragment of Puma promoter was cloned into the pGL3-basic luciferase reporter vector. The biological activity of Pumareporter plasmid was verified by cell transfection. Methods The target fragments of Puma were amplified by RT-PCR method and the fragments were inserted into the pGL3-basic luciferase reporter vector. The acquired Puma-Luc plasmid was transfected into H1299 cell line and detected its activity. Results Sequencing indicated that the amplified Puma promoter is correct. Dual-luciferase Reporter Assay showed the Puma-Luc constructs have promoter activity. Conclusion The cloning of human Puma gene promoter and the construction of its reporter vector were successful. This study will lay the foundation for further research on the function of p53 inducing apoptosis through mitochondrial pathway.
4.Discussion about risk and management of Chinese patent medicine with double identity.
Yun JIN ; Sai-wei WU ; Xing-wang GAO ; Dan-hua XU ; Yong-song ZHAI
China Journal of Chinese Materia Medica 2015;40(11):2249-2251
Chinese patent medicine with double identity was a special phenomenon, and many preparations not only were prescription drugs but also over the counter ( OTC) drugs, which brought a lot of trouble. Based on statistics of list of OTC medicines of CFDA, related varieties, route of administration and functions of these drugs were searched. The causes of insufficient were analyzed and the potential risk was investigated. To ensure the safety of drug usage for the patient, risk management system should be set up by improving the technical requirements for registration, improving the drug labels and manuals, playing the role of pharmacists in pharmacy services and raising awareness of doctor and patient for these drugs.
China
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Humans
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Nonprescription Drugs
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adverse effects
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Risk Management
5.Influence of Four Kinds of Material on the Results of Two Kinds of D-dimer Detection Reagent Assay
Yun ZHANG ; Min HUANG ; Xing GAO ; Min LIU ; Nuohan LIU ; Juming BAI ; Qiangyuan ZHAO
Journal of Modern Laboratory Medicine 2014;(5):164-166
Objective To observe the influence of triglyceride(TG),hemoglobin(HGB),total bilirubin(TBIL)and rheumatoid factor(RF)on the results of two kinds of D-dimer detection reagent assay.Methods Five samples were prepared into series with 2,4,6,8 and 10 mmol/L triglyceride by adding fat emulsion.Five samples were prepared into series with 1,2,3,4 and 5 g/L of hemolysis degree by adding hemoglobin solution.Five samples were prepared into series with 20,40,60,80 and 100μmol/L total bilirubin by adding bilirubin standard preparation.Ten samples were prepared into rheumatoid factor levels be-tween 0~150 IU/L by adding rheumatoid factor standard solution.Simultaneously with D-dimer detection reagent and D-di-mer HS detection reagent for testing,each sample was measured two times and the results averaged.Results When TG≤2 mmol/L,the D-dimer reagents without interference,and TG≥4 mmol/L,the D-dimer reagents due to “SD baseline data out of range”couldn’t be detected.TG≤10 mmol/L,for D-dimer HS reagents was without interference.When HGB≥1 g/L, the D-dimer reagents due to“SD baseline data out of range”couldn’t be detected.HGB≤4 g/L,for D-dimer HS reagents was without interference.When hemoglobin levels was equal to 5 g/L,the D-dimer HS reagents test results false increased about 30.4%.With total bilirubin concentration increases,D-dimer and D-dimer HS reagents test results were false increase, showed amplitude by a power law,and the two reagents increased consistency.Preparation a series of pooled plasma of rheu-matoid factor levels in 0~150 IU/L,D-dimer test results reagent falsely elevated levels of rheumatoid factor with a linear correlation (Y=59.31X+50.43,R2=0.998).When RF≤150 IU/L,the D-dimer HS reagents was without interference. Conclusion Triglyceride,hemoglobin,total bilirubin and rheumatoid factor may interfere D-dimer testing process,icterus and severe hemolysis will interfere D-dimer HS testing process.D-dimer HS reagents against interfere exceed D-dimer rea-gents.When D-dimer test results does not match with clinicians determine,the influence ofinterfering substances should be considered.
7.The Application of Heating Effect in Breeding of Microorganism
Xing-Qiang GAO ; Yun-Hong HUANG ; Fei DAI ; Xue-Qin FU ; Zhong-Er LONG ;
Microbiology 2008;0(10):-
Advances in mechanism and application of the heating effect in breeding of microorganism are reviewed in this paper. Heat produces mutagenesis effect and screening effect. Heating mutagenesis effect is occurred through the substitution of G-C base pair induced by heat, and heating screening effect produces higher forward mutation rate induced by other mutagens.
8.A phytochemical and pharmacological advance on medicinal plant Litsea cubeba (Lauraceae).
Shui-Ying ZHANG ; Qiang GUO ; Xiao-Li GAO ; Zhi-Qin GUO ; Yun-Fang ZHAO ; Xing-Yun CHAI ; Peng-Fei TU
China Journal of Chinese Materia Medica 2014;39(5):769-776
Litsea cubeba is one of aromatic medicinal plant belonging to family Lauraceae. The roots, stems and fruits of L. cubeba have been widely applied as folk medicines in some districts in China for relieving rheumatism and cold, regulating Qi (meridian) to alleviate pain. Previous studies revealed that this species contains major alkaloids, in specific aporphines, and minor flavonoids, lignans as well. Related pharmacological investigations demonstrated its activities and clinical applications on cardiovascular diseases, anti-cancer, against rheumatoid arthritis, relieving asthma and anti-allergic effects, as anti-oxidants, and so on. As an effort for further exploration of this bioactive ingredients and potential drug development, this paper summarizes most phytochemical and pharmacological results. Further, future prospects are also included.
Animals
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Drug Therapy
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Humans
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Litsea
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chemistry
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Molecular Structure
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Plant Extracts
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chemistry
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pharmacology
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Plants, Medicinal
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chemistry
9.Effects of Bushen Wenyang Huayu Recipe on Expressions of HIF-1α, PHD2, and VHL in Endometriosis Rats with Shen Yang Deficiency Blood Stasis Syndrome.
Yun-bo JIA ; Hui-lan DU ; Xing GAO ; Wen-hui BIAN ; Xiao-hua LIN ; Guang-guo BAN ; Qian-hua TIAN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(10):1210-1217
OBJECTIVETo observe the effect of Bushen Wenyang Huayu Recipe (BWHR) on hypoxia inducible factor-1α (HIF-1α), proline hydroxylase2 (PHD2), von Hippel Lindau disease (VHL) suppressor gene expressions in endometriosis (EM) rats with Shen yang deficiency blood stasis syndrome (SYDBSS), and to explore the pathogenesis of EM and the mechanism of BWHR for treating EM.
METHODSTotally 50 SD rats were randomly divided into five groups, i.e., the blank control group, the sham-operation group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 in each group. Rats in the blank control group and the sham-operation group were fed routinely. Rats in the rest 3 groups received 30-day "extended refrigerator freezing and ice water immersion" and combined with " autotransplantation" to establish EM rat model with SYDBSS. One Milliliter BWHR at 3.33 g/mL was administered to rats in the CM group by gastrogavage. Gestrinone at the daily dose of 0. 5 mg/kg was administered to rats in the WM group by gastrogavage. Equal volume of normal saline was administered to rats in the model group, the blank control group, and the sham-operation group. The size and morphology of ectopic foci in rats were observed after 4 weeks of medication. Expressions of serum CA125, plasma cyclic adenosine monophosphate (cAMP), and plasma cyclic guanosine monophosphate (cGMP) were detected by radioimmunoassay. Morphological changes of eutopic endometrium and ectopic tissue were observed under the optical microscope by HE staining. Protein expressions and contents of HIF-lα, PHD2, and VHL were detected by immunohistochemical SABC method and Western blot. mRNA expressions of HIF-1α, PHD2, and VHL were detected by RT-PCR.
RESULTSThe ectopic foci grew significantly in the model group. Their volumes were obviously contracted after treated by CM and WM. Compared with the blank control group and the sham-operation group, serum CA125 and plasma cGMP obviously increased, cAMP obviously decreased (P < 0.05); expressions and contents of HIF-1α mRNA and protein all decreased (P < 0.05); mRNA and protein expressions and contents of PHD2 and VHL all decreased in the model group (P < 0.05). Compared with model group, levels of CA125 and cGMP obviously decreased; cAMP levels obviously increased, expressions and contents of HIF-1α mRNA and protein all increased, mRNA and protein expressions and contents of PHD2 and VHL all increased in the WM group and the CM group (P < 0.05). Compared with the CM group, PHD2 protein contents were higher in the WM group (P < 0.05). HIF-1α was negatively correlated with PHD2 (r = -0.799, P = 0.00). HIF-1α was negatively correlated with VHL (r = -0. 625, P = 0.003).
CONCLUSIONSBWHR could effectively treat EM. Its mechanism might be associated with reducing contents of HIF-1α, serum CA125, and plasma cGMP, and up-regulating expressions of PHD2, VHL, and cAMP.
Animals ; Cyclic AMP ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; drug therapy ; metabolism ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Proline ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Up-Regulation ; Yang Deficiency ; drug therapy ; metabolism
10.Effects of 1,4-benzoquinone on the proliferation activity of human bone marrow stem cells.
Yun XIAO ; Li JU ; Wei WU ; Xiang-li GAO ; Jing WANG ; Xing ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(5):343-347
OBJECTIVETo explore the influence of 1,4-benzoquinone (1,4-BQ) on proliferation of human bone marrow haematopoietic stem cells (hBM-HSCs) and human bone marrow mesenchymal stem cells (hBM-MSCs).
METHODSThe bone marrow samples were collected from a healthy donor. Methylcellulose semi-solid culture medium was used to culture the mononuclear cells of bone marrow in different culture systems. Colony-forming unit (CFU) assay was utilized to evaluate the proliferation of hBM-HSCs exposed to 1,4-BQ at the doses of 10, 25, 50 and 100 µmol/L and to observe the influence of 1,4-BQ on the Colony-forming unit-erythroid (CFU-E)/Burst-forming unit-erythroid (BFU-E), Colony-forming unit-granulocyte, macrophage (CFU-GM), Colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-GEMM) in hBM-MSCs. MTT assay was used to detect the proliferation of hBM-MSCs exposed to 1,4-BQ at the doses of 1, 5, 10, 25, 50, 100, 200, 500 and 1000 µmol/L for 24 h, respectively, after hBM-MSCs were isolated, cultured and expanded.
RESULTSThe results of CFU assay indicated that numbers of CFU-E/BFU-E, CFU-GM and CFU-GEMM in 25, 50 and 100 µmol/L groups significantly decreased, as compared with control group (P < 0.05). However, no significant difference was found between the 10 µmol/L group and the control group. The results of MTT assay showed that the cellular viability of hBM-MSCs exposed to 1,4-BQ at the doses of 50 ∼ 200 µmol/L for 24 h significantly decreased in a dose-depended manner. When the exposure dose was higher than 200 µmol/L, the cellular viability of hBM-MSCs was lower than 5% which was significantly lower than that of control group (P < 0.05). When the exposure dose was lower than 25 µmol/L, there was no significant difference of cellular viability between exposure group and control group (P > 0.05).
CONCLUSIONThe results of the present study demonstrated that 1,4-BQ could inhibit the colony forming of hBM-HSCs and the relative viability of hBM-MSCs in vitro. The hematotoxicity induced by 1,4-BQ may be related to inhibiting the proliferation capacity of hBM-HSCs.
Benzoquinones ; toxicity ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Erythroid Precursor Cells ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology