Effects of acupuncture on pulmonary function and airway smooth muscle spasm in asthma rats / 中国针灸

OBJECTIVE@#To observe the effects of acupuncture at "Kongzui" (LU 6) and "Yuji" (LU 10) on the latent period of inducing asthma, pulmonary function and the expression of endothelin-1 (ET-1) and metallothionein-2 (MT-2) in asthma rats, and to explore the possible mechanism of acupuncture in alleviating airway smooth muscle spasm and improving the acute attack of asthma.@*METHODS@#A total of 40 male SD rats of SPF-grade were randomly divided into a normal group, a model group, a medication group and an acupuncture group, 10 rats in each group. Except for the normal group, ovalbumin sensitization method was used to establish the asthma model in the other 3 groups. Salbutamol nebulization was adopted in the medication group, while acupuncture was applied at bilateral "Kongzui" (LU 6) and "Yuji" (LU 10) in the acupuncture group. The intervention was given once a day for 14 days in the two groups. The latent period of inducing asthma and pulmonary function were observed, the levels of ET-1 and tumor necrosis factor (TNF)-α in serum and bronchoalveolar lavage fluid (BALF) were detected by ELISA method, the morphology of the airway was observed by Masson staining, the ultrastructure of the airway smooth muscle was observed by transmission electron microscopy, the mRNA and protein expression of ET-1 and MT-2 in lung tissue was detected by real-time PCR and Western blot methods.@*RESULTS@#Compared with the normal group, in the model group, the latent period of inducing asthma was shortened (P<0.01); the airway resistance (RL) was increased while the dynamic compliance (Cdyn) was decreased (P<0.01, P<0.05); the levels of ET-1 and TNF-α in serum and BALF were increased (P<0.01); collagen fibers and collagen depositions were found around the bronchi, airway smooth muscle was thickened, the cell damage was severe and mitochondria were swollen; the mRNA and protein expression of ET-1 was increased while the mRNA and protein expression of MT-2 was decreased (P<0.01). Compared with the model group, in the acupuncture group, the latent period of inducing asthma was prolonged (P<0.05), the RL was decreased while the Cdyn was increased (P<0.01, P<0.05). Compared with the model group, in the medication group and the acupuncture group, the levels of ET-1 and TNF-α in serum and BALF were decreased (P<0.01, P<0.05); collagen fibers and collagen depositions around the bronchi were reduced, the thickened airway smooth muscle was lightened, the cell damage was improved; the mRNA and protein expression of ET-1 was decreased while the mRNA and protein expression of MT-2 was increased (P<0.01). Compared with the medication group, the mRNA expression of MT-2 was increased in the acupuncture group (P<0.05).@*CONCLUSION@#Acupuncture at "Kongzui" (LU 6) and "Yuji" (LU 10) can improve the pulmonary function and alleviate the airway smooth muscle spasm in rats with asthma. Its mechanism may be related to the down-regulation of ET-1 expression and up-regulation of MT-2 expression.

Genome-wide analysis of metallothionein gene family in maize to reveal its role in development and stress resistance to heavy metal

BACKGROUND: Maize (Zea mays L.) is a widely cultivated cereal and has been used as an optimum heavy metal phytoremediation crop. Metallothionein (MT) proteins are small, cysteine-rich, proteins that play important roles in plant growth and development, and the regulation of stress response to heavy metals. However, the MT genes for maize have not been fully analyzed so far. METHODS: The putative ZmMT genes were identified by HMMER. The heat map of ZmMT genes spatial expression analysis was generated by using R with the log2 (FPKM + 1). The expression profiles of ZmMT genes under three kinds of heavy metal stresses were quantified by using qRT-PCR. The metallothionein proteins was aligned using MAFFT and phylogenetic analysis were constructed by ClustalX 2.1. The protein theoretical molecular weight and pI, subcellular localization, TFs binding sites, were predicted using ProtParam, PSORT, PlantTFDB, respectively. RESULTS: A total of 9 ZmMT genes were identified in the whole genome of maize. The results showed that eight of the nine ZmMT proteins contained one highly conserved metallothio_2 domain, while ZmMT4 contained a Metallothio_PEC domain. All the ZmMT proteins could be classified into three major groups and located on five chromosomes. The ZmMT promoters contain a large number of hormone regulatory elements and hormone-related transcription factor binding sites. The ZmMT genes exhibited spatiotemporal specific expression patterns in 23 tissues of maize development stages and showed the different expression patterns in response to Cu, Cd, and Pb heavy metal stresses. CONCLUSIONS: We identified the 9 ZmMT genes, and explored their conserved motif, tissue expression patterns, evolutionary relationship. The expression profiles of ZmMT genes under three kinds of heavy metal stresses (Cu, Cd, Pb) were analyzed. In summary, the expression of ZmMTs have poteintial to be regulated by hormones. The specific expression of ZmMTs in different tissues of maize and the response to different heavy metal stresses are revealed that the role of MT in plant growth and development, and stress resistance to heavy metals.

Cell-penetrating Effects and Repair of Cell Oxidative Damage of TAT-metallothionein Fusion Proteins / 中国生物化学与分子生物学报

The human Immunodeficiency Virus Transactivator (TAT) protein transduction peptide is a trans-transcription activator encoded by HIV-1. It is rich in basic amino acids, and capable of efficiently mediating the passage of exogenous macromolecules through a variety of membrane structures, such as the cytoplasmic membrane and the blood-brain barrier. Metallothionein (MT) is a protein with low molecular weights and rich cysteine contents. It plays important roles in maintaining the dynamic balance of metal contents in the body, in the detoxification of heavy metals and in defense against oxidative stress. Based on the full-length MT cDNA previously cloned from Sinopotamon henanense, we aim to prepare a TAT-mediated recombinant fusion protein that can cross the membrane and enter the cell by means of genetic engineering. The hydroxyl radical scavenging rate and total antioxidant capacity of TAT-MT were measured in vitro. An immunofluorescence technique was used to detect the transmembrane activity. An MTT assay was used to study the repair effect of H

Immunoreactive Scoring of Metallothionein-3 (MT-3) Expression as an Indicator of Metallic Taste Rats which are Induced by Doxorubicin and Antioxidant

@#Introduction: Cancer is one of the leading causes of deathworldwide. Chemotherapy like as doxorubicin is the most common treatment procedure given to cancer patients. Doxorubicin is a cytotoxic drug that triggers the production of Reactive Oxygen Species (ROS) affect to body cells including taste bud cells to induced the expression of Metallothionein 3 (MT-3), eventually cause cell damage that leads to a metallic taste. Antioxidant therapy can be an alternative to overcome metallic taste as it counters ROS effect and lowers the expression of MT-3. The aim of this study is to evaluate MT-3 expression in the taste bud cells of male Wistar Rats after induction of doxorubicin combined with vitamin E and mung bean sprouts (Phaseolusradiatus L.) juice. Methods: 27 male Wistar rats weighing 250-300 g aged 3-4 months were divided into 3 groups randomly; control group, treatment group 1 (receiving doxorubicin and vitamin E), and treatment group 2 (receiving doxorubicin and mung bean sprouts juice). After 5 days, the rats were sacrificed, and the tongue was taken for immunohistochemistry analysis. Data were then analyzed by One Sample Kolmogorov-Smirnov, Levene Test, and Oneway-ANOVA statistical test (p<0.05). Results: The MT-3 expression increases in the following order; control group (4.93), treatment group 2 (7.08), and treatment group 1 (9.95). Treatment group 1 and 2 both show remarkable increases of MT-3 expression compare to control. Conclusion: The induction of doxorubicin and antioxidant can increase the level of MT-3 expression.

Metallothionein-2a (rs1610216&rs28366003) gene polymorphisms and the risk of stomach adenocarcinoma / Polimorfismos do gene metalotioneína-2A (rs1610216& rs28366003) e o risco de adenocarcinoma de estômago

ABSTRACT BACKGROUND: Gastric cancer is the fourth most common cause of worldwide cancer. Also in contrast to the huge advances in curing, the chance of living is very low even in surgery cases. Having a genetic predisposition plays an important role in cancer development. The association between Metallothionein-2A gene polymorphisms and the risk of adenocarcinoma has been widely studied, yet there is only one study on stomach diseases. OBJECTIVE: In this study, we aimed to investigate the association between 2 (MT-2A) polymorphisms and adenocarcinoma. METHODS: This cross-sectional case control study was performed between Mach 2014 and January 2015 at the Tuba Hospital of Sari, Iran. Peripheral blood samples were collected in EDTA tube. DNA extraction was performed using the spin column procedure. The MT-2A polymorphisms MT-2A (rs1610216), (rs28366003) were determined by polymerase chain reaction-restriction fragment length polymorphism analysis in 95 a topic adenocarcinoma patients and 90 healthy individuals from Iranian population. RESULTS: The MT-2A rs1610216 polymorphism increased the risk of adeno carcinoma in our Iranian population [OR: 3.8533; 95%CI, 1.3155-11.2869; P=0.0139] and rs28366003 [OR: 4.0978; 95%CI, 1.2521-13.4108; P=0.0197]. CONCLUSION: The MT-2A gene polymorphism was associated with the risk of adenocarcinoma in the Iranian population.

RESUMO CONTEXTO: O câncer gástrico é a quarta causa mais comum de câncer em todo o mundo. Também em contraste com os enormes avanços na cura, a chance de viver é muito baixa, mesmo em casos de cirurgia. Ter uma predisposição genética desempenha um papel importante no desenvolvimento do câncer. A associação entre polimorfismos do gene metalotioneína-2A e o risco de adenocarcinoma tem sido amplamente estudada, mas há apenas um estudo sobre doenças estomacais. OBJETIVO: Neste estudo, objetivou-se investigar a associação entre 2 (MT-2A) polimorfismos e adenocarcinoma. MÉTODOS: Um estudo de controle de caso transversal foi realizado entre março de 2014 e janeiro de 2015 no hospital Tuba, Sari, Irã. Amostras de sangue periférico foram coletadas em tubo EDTA. A extração do ADN foi executada usando o procedimento da coluna da rotação. Os polimorfismos MT-2a MT-2A (rs1610216), (rs28366003) foram determinados pela análise do polimorfismo do comprimento do fragmento da reação-limitação de cadeia da polimerase em 95 pacientes com adenocarcinoma tópico e em 90 indivíduos saudáveis da população iraniana. RESULTADOS: O polimorfismo MT-2A rs1610216 aumentou o risco de adenocarcinoma de em nossa população iraniana. [OR: 3,8533; 95%CI, 1,3155-11,2869; P=0,0139] e rs28366003 [OR: 4,0978; 95%CI, 1,2521-13,4108; P=0,0197]. CONCLUSÃO: O polimorfismo do gene MT-2A foi associado ao risco de adenocarcinoma na população iraniana.

Immunohistochemical Expression of Oxidative Stress Markers in Bovine Tissues Correlated to Cadmium Concentration

A study was conducted to assess oxidative stress in bovine tissues specifically liver, kidney and lungs by using immunohistochemistry (IHC) and their correlation to cadmium (Cd) toxicity in bovine. Metallothionein and malondialdehyde are oxidative stress markers and their expression increases as cadmium concentration increases in tissues. In present study, a total of 62 bovine tissue samples were randomly collected from the animals found dead near industrial or expected polluted areas of Jabalpur city after detailed postmortem examination. These samples were processed for cadmium estimation and IHC staining in tissues. 200 mg of tissue samples were acid digested and cadmium concentration were estimated by using ICP-OES. In our study, cadmium concentration in tissues was ranged from 0.040 to 3.952 ppm in liver, 0.050 to 3.949 ppm in kidney and 0.020 to 3.134 ppm in lungs of bovine. These samples further grouped under three groups with cadmium level 0-1, 1-2 and > 2 ppm, according to Puls criteria. Majority of samples had cadmium level in the range of 0-1 ppm. Approximately 8, 13, 5% liver, kidney and lung tissues respectively had cadmium concentration > 2 ppm are considered under high risk. Formalin fixed and paraffin processed representative samples of liver, kidney and lungs were stained immunohistochemically by using commercially available antibodies for metallothionein and malondialdehyde protein. Tissues with high cadmium level showed increase in expression of metallothionein in nucleus and cytoplasm of the tissues along with increased cytoplasmic expression of malondialdehyde in liver, kidney and lung tissues as compared to low cadmium level

Maternal zinc metabolism and changes of metallothionein-1 and zinc transporter-1 in fetal congenital heart diseases / 中华临床营养杂志

Objective To investigate maternal zinc metabolism and the changes of zinc-related factors as metallothionein-1 (MT1) and zinc transporter-1 (ZnT1) in certain types of congenital heart diseases (CHD).Methods Fifteen infants with interventricular septal defect,12 infants with atrial septal defect and 7 infants with tetralogy of Fallot,together with their mothers were enrolled,and normal infants and their mothers were enrolled by a ratio of 1 ∶ 1 with the above three types of CHD diseases.General conditions of the mothers,along with their diets and zinc-containing drug supplementation during the pregnancy,were surveyed.Maternal blood zinc levels and serum alkaline phosphatase activities at gestation week 32 and delivery or induced abortion,and the protein and mRNA expressions of MT1 and ZnT1 in maternal serum and placental tissue at delivery or induced abortion were assayed.Results The general conditions were comparable between the CHD group and control group.The ratio of the mothers taking more zinc-rich food was significantly lower in the CHD group than in the control group.Circulating zinc levels in interventricular septal defect (73.55±5.79 μmol/L),atrial septal defect (72.66±5.82 μmol/L) and tetralogy of Fallot (68.72±6.72 μmol/L) groups were significantly lower than those in the control groups (82.77± 7.88,84.58 ± 7.55 and 85.66 ± 7.30 μmol/L) at delivery (P all ＜ 0.05).Similar change patterns were seen for serum alkaline phosphatase activities.The relative quantities of serum MT1 and ZnT1 proteins in interventricular septal defect (73.22±36.54 and 68.55± 27.82),atrial septal defect (64.29± 38.26 and 74.55 ± 29.67) and tetralogy of Fallot (67.88± 30.50 and 70.13±29.65) groups were significantly lower than those in their corresponding control groups (166.31±67.43and 97.67±30.22,182.56±71.40 and 111.65±32.70,and 173.81±62.36 and 108.27±28.52,P＜0.01 or P＜0.05).The relative quantities of placental MT1 and ZnT1 proteins and mRNA expressions in interventricular septal defect (protein quantities 0.438±0.096 and 0.384±0.061,mRNA expressions 1.23±0.82 and 0.96±0.39),atrial septal defect (0.427±0.093 and 0.377±0.059,1.17±0.70 and 0.85±0.40) and tetralogy of Fallot (0.414±0.111 and 0.336±0.066,1.31±0.97 and 0.90±0.38) groups were significantly lower than those in their corresponding control groups (protein quantities 0.565±0.083 and 0.541±0.090,mRNA expressions 2.78± 1.06 and 1.67±0.33;protein quantities 0.622±0.136 and 0.493±0.079,mRNA expressions 2.85±0.89 and 1.72±0.38;protein quantities 0.637±0.125 and 0.521±0.089,mRNA expressions 3.21 ± 0.99 and 1.61±0.29;P＜0.01 or P＜0.05).Conclusion Mothers with their fetus of certain types of CHD are found zinc deficiency,and down-regulation of MT1 and ZnT1 expressions in the serum and placenta may involve in the pathogenesis of CHD when maternal zinc deficiency.

Correlation of metallothionein-2A, E-cadherin and cyclin E with biochemical recurrence in prostate cancer / 肿瘤研究与临床

Objective To investigate the expressions of metallothionein-2A (MT-2A), E-cadherin, interleukin-6 (IL-6), cyclin E, proliferating cell nuclear antigen (PCNA) and bcl-2 in prostate cancer tissues and their correlation with biochemical recurrence of prostate cancer. Methods Tissue specimens from 128 cases of prostate cancer who underwent radical prostatectomy in Shanxi Dayi Hospital from October 2012 to October 2017 were processed and transferred into tissue microarrays, the clinicopathological parameters of patients were also recorded. The expression levels of MT-2A, E-cadherin, IL-6, cyclin E, PCNA and bcl-2 were detected by immunohistochemical avidin-biotin complex (ABC) staining. The correlation between different molecular markers and biochemical recurrence of prostate cancer was analyzed. Results The biochemical recurrence rate of 128 patients with prostate cancer was 30.5％ (39/128). The biochemical recurrence rates of low-risk, intermediate-risk and high-risk prostate cancer patients were 14.8％(8/54), 38.7％(24/62) and 58.3％ (7/12), respectively. The risk classification and pathological T stage of patients with prostate cancer were associated with the expressions of MT-2A, cyclin E, IL-6 and E-cadherin (all P< 0.05). Multivariate Cox risk model showed that the high risk classification (HR= 1.81, 95％CI 1.56-2.19, P=0.042), MT-2A positive expression (HR= 2.01, 95％CI 1.08-3.15, P= 0.005), cyclin E positive expression (HR= 1.79, 95％CI 1.08-2.21, P= 0.042) and E-cadherin negative expression (HR= 1.92, 95％ CI 1.22-2.45, P= 0.020) were the independent risk factors for biochemical recurrence of prostate cancer. Conclusion The expression of MT-2A, cyclin E and E-cadherin may serve as independent predictors for biochemical recurrence of prostate cancer.

Enrichment and Identification of Metallothionein by Functionalized Nano-Magnetic Particles and Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry / 分析化学

Metallothionein ( MT) is a low-molecular-weight protein with high inducibility and binding ability with metal ions. Therefore, MT is often regarded as an important biomarker for assessment of heavy metal pollution in water environment. But the traditional process of its enrichment and identification is time-consuming and complicated. Herein, we prepared a core-shell nanoparticle, gold-coated iron oxide nanoparticles ( Fe3O4@Au NPs) . The nanoparticle possessed the advantages such as fast response to magnetic fields and optical properties attributing to Fe3O4and Au nanoparticles separately. Fe3O4@Au nanoparticles were used to enrich MT simply through Au-S interaction, and the purified proteins were determined by matrix assisted laser desorption ionization time-of-flight mass spectrometry ( MALDI-TOF/MS) . The results in this work showed that the Fe3O4@Au nanoparticles could directly enrich MT from complex solutions and the detection limit could be down to 10 fg/mL.

Heterologous expression of a rice metallothionein isoform (OsMTI-1b) in Saccharomyces cerevisiae enhances cadmium, hydrogen peroxide and ethanol tolerance

Abstract Metallothioneins are a superfamily of low-molecular-weight, cysteine (Cys)-rich proteins that are believed to play important roles in protection against metal toxicity and oxidative stress. The main purpose of this study was to investigate the effect of heterologous expression of a rice metallothionein isoform (OsMTI-1b) on the tolerance of Saccharomyces cerevisiae to Cd2+, H2O2 and ethanol stress. The gene encoding OsMTI-1b was cloned into p426GPD as a yeast expression vector. The new construct was transformed to competent cells of S. cerevisiae. After verification of heterologous expression of OsMTI-1b, the new strain and control were grown under stress conditions. In comparison to control strain, the transformed S. cerevisiae cells expressing OsMTI-1b showed more tolerance to Cd2+ and accumulated more Cd2+ ions when they were grown in the medium containing CdCl2. In addition, the heterologous expression of GST-OsMTI-1b conferred H2O2 and ethanol tolerance to S. cerevisiae cells. The results indicate that heterologous expression of plant MT isoforms can enhance the tolerance of S. cerevisiae to multiple stresses.

Time-dependent Expression of MT1A mRNA and MT2A mRNA in the Contused Skeletal Muscle of Rats / 法医学杂志

OBJECTIVES@#To investigate the time-dependent expression of metallothionein （MT） 1A mRNA and MT2A mRNA in contused skeletal muscle of rats.@*METHODS@#A total of 54 Sprague-Dawley rats were used in this study. The rats were divided into two parts: control group （n=6） and contusion groups （0.5, 1, 6, 12, 18, 24, 30, and 36 h after contusion, n=6）. Total RNA was extracted from skeletal muscle. The expression levels of MT1A mRNA and MT2A mRNA were detected by SYBR Green I real-time PCR.@*RESULTS@#The expression trends of the two potential marker genes were related to wound age. In addition to 0.5 h, there were significant contrasts between the control group and contused group （P<0.05）, about the expression levels of MT1A mRNA and MT2A mRNA in different phases. As the extension of wound age, the relative expression of MT1A mRNA and MT2A mRNA at 1 h, 6 h, 12 h and 18 h after contusion demonstrated upgrade tendency until its expression levels in 18 h peak with 239.41±15.20 and 717.42±50.76, respectively. When time extends to 24 h after injury, the expression of above two marks decreased, respectively. The MT1A mRNA and MT2A mRNA expression levels increased at 30 h and then decreased.@*CONCLUSIONS@#Determination of MT1A mRNA and MT2A mRNA levels by real-time PCR may be useful for the estimation of wound age.

Expression of metallothionein 1H in children and adolescents osteosarcoma and effect on cell proliferation / 中华实用儿科临床杂志

Objective To detect the expression levels of metallothionein1 H(MT1 H)in children and adoles-cents osteosarcoma serums,and to analyze its relationship with clinicopathological features,and to explore the effect of MT1 H on cell proliferation of osteosarcoma cells and its mechanism.Methods Enzyme -linked immuno sorbent assay (ELISA)was performed to detect the expression of MT1 H in children and adolescents osteosarcoma serums and non-neoplastic disease serums.MT1 H vector was transfected into the osteosarcoma U2OS cells.Reverse transcription -poly-merase chain reaction(RT -PCR)and Western blot were used to detect the expression of the mRNA and protein of MT1 H,respectively.Methylthiazolyldiphenyl -tetrazolium bromide(MTT)was used to detect the cell growth.Western blot was performed to detect the expression of nuclear factor(NF)-κB,and inhibitor of κB (IκB)-αprotein. Results The expressions of MT1 H in osteosarcoma serums and nonneoplastic disease serums was (0.51 ± 0.52)μg/L and (2.17 ±0.78)μg/L,respectively,with a significant difference between the 2 groups(t =-8.966, P <0.05).The expression of MT1 H in stage Ⅰ -ⅡA andⅡB -Ⅲ was (1 .98 ±0.69)μg/L and (2.45 ±0.82)μg/L,respectively,showing a gradual increase depending on clinical staging(t =-2.343,P <0.05).The expressions of MT1 H mRNA and protein were elevated in osteosarcoma U2OS cells after MT1 H vector transfection(all P <0.05). MTT assay showed that,the A value in blank control group,blank vector group,MT1 H vector group were 0.38 ±0.03, 0.36 ±0.03,0.42 ±0.03,respectively,the cell proliferation in the MT1 H vector group was significantly promoted when compared with these in the blank vector group and blank control group(F =4.213,P <0.05)from the third day.West-ern blot showed that,the relative expression of NF -κB in blank control group,blank vector group,MT1 H vector group were 0.56 ±0.05,0.53 ±0.05,0.92 ±0.07,respectively,the relative expression of IκB -αprotein were 0.64 ± 0.06,0.62 ±0.09,0.34 ±0.08,respectively,the expression of NF -κB protein was up -regulated and the expression of IκB -αprotein was down -regulated in the MT1 H vector group when compared with those in the blank vector group and blank control group(F =44.581 ,14.927,all P <0.05).Conclusions The expression of MT1 H is increased in children and adolescents osteosarcoma serums compared with that in nonneoplastic disease serums.The clinical stage is later,the expression of MT1 H is higher.MT1 H promotes cell proliferation through regulating the NF -κB pathway.

Association of plasma metallothionein 3 and its polymorphisms with childhood autism / 中华实用儿科临床杂志

Objective To investigate the association of plasma metallothionein 3 (MT3) and its polymorphisms with childhood autism,in order to provide the objective evidence for autistic etiology and molecular diagnosis.Methods A total of 132 autistic children were recruited from several special autism training schools in Wuhan and the Hubei Maternal and Child Health Hospital between January 2011 and November 2014.Three hundred and sixteen healthy children from the out-patients of Zhongnan Hospital of Wuhan University during the same period were enrolled as healthy controls.Enzyme linked immunosorbent assay was utilized to measure plasma MT3 protein levels in a dataset of 81 cases and 80 controls,while eight single nucleotide polymorphisms (SNP) located in MT3 gene were genotyped in another greater dataset that included 132 cases and 236 controls by the matrix-assisted laser desorption/ionization time of flight mass spectrometry within the Sequenom platform.Results Plasma MT3 protein level was significantly lower in autistic group compared to healthy controls [(740.0 ± 327.4) ng/L vs (1 007.1 ± 554.3) ng/L,P ＜ 0.001],particularly for boys when stratified by gender (P =0.005).No difference existed in any allele or genotype frequencies between the 2 groups (all P ＞ 0.05).Conclusions The selected autistic children harbored abnormal expression profiles of plasma MT3 protein,which may have no connection with its gene polymorphisms.

Difference of Metallothionein Gene G-201A Polymorphism in Type 2 Diabetes and Essential Hypertension / 昆明医科大学学报

Objective To investigate the difference of MT4 G-201A of Metallothionein (metallotionein, MT) family gene locus single nucleotide polymorphisms (SNPs) in type 2 diabetes and essential hypertension. Mothods We selected 324 cases of patients with type 2 diabetes, 301 cases of patients with essential hypertension, 301 case of normal physical examination population. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology was used to detect MT4 G-201A polymorphism. Results (1) The genotype GG, GA and AA frequency of MT4 G-201A in 324 cases of patients with type 2 diabetes,301 cases of patients with essential hypertension, 301 case of normal physical examination population were 39.5%, 46.6%, 13.9%;42.2%, 48.5%, 9.3%;and 42.5%, 51.5%, 6.0%, respectively. The genotypes in all the distribution were in Hardy-Weinberg equilibrium (P>0.05) . (2) MT4 G-201A polymorphism:Three groups of genotype and allele frequency distribution (G and A) had differences (P0.05);there was a difference between type 2 diabetes group and the normal group (P0.05) . (3) MT4 G-201A polymorphism in Men:Three groups of genotype and allele frequency distribution had differences (P0.05) .MT4 G-201A polymorphism between women:Three groups of genotype and allele frequency distribution had differences (P<0.05), there was a difference between type 2 diabetes and normal group genotypes (P<0.05) . Conclusions (1) There is correlation of MT gene G-210A polymorphism and type 2 diabetes, no association with essential hypertension. (2) There is a difference of MT gene G-210A polymorphism in type 2 diabetes in women, no difference in men.

Effect of Raloxifene on Cadmium Chloride-Contaminated Renal Tubular Epithelial Cells / 医药导报

Objective To investigate the effect of raloxifene on ERα, ERβ and MT-1a expression in renal tubular epithelial cells contaminated with cadmium chloride,for exploring the protective effect and action mechanism of raloxifene in renal injury induced by cadmium. Methods Renal tubule cells were isolated from kidneys of new born SD rats and purified by Percoll. The cells of the second generation were selected for experiment. The cells were divided into 3 groups:blank control group,cadmium chloride group and raloxifene group. After 4 h,cell viability was detected by MTT,and after 24 h,mRNA and protein expression levels of ERα, ERβ and MT-1a in renal tissues were determined by RT-PCR and immunohistochemistry technology,respectively. Results Compared with the blank control group,morphology of plenty cells were changed in cadmium chloride group,a number of cells were dead,and the survival rate was only 55.3%;at the same time,mRNA and protein expression levels of ERβ and MT-1a significantly increased (P0.05) . Conclusion Raloxifene can bond with ERβcompetively,down-regulate MT-1a and decrease Cd-MT synthesis,so as to reduce cadmium-induced damage of renal tubular epithelial cells.

Antagonizing effects of Metallothionein against development of skin cancer and relevant mechanisms / 中国免疫学杂志

Objective:To establish a UVB damage cell model with HaCaT cells to investigate the protective effects of Zinc sulfate on the cell damage caused by UVB and its relevant mechanisms .Methods: The cells were divided into normal group , Zinc group,UVB group,Znic and UVB group.The addition of Zinc sulfate to the HaCaT cells was conducted 24h prior to the irradiation to the cells by UVB.Cell apoptosis was detected by Western blot and the expression of metallothionein and NF -κB/p65 were measured by im-munohistochemistry.Results:Compared with normal and Zn+UVB group, Bax/Bcl-2 rate in UVB group increased.Compared with normal group ,MT expression levels in UVB group ,Zn group increased ,and compared with UVB group ,MT expression level in Zn+UVB group increased .Compared with normal group and Zn+UVB group,NF-κB/p65 expression level in UVB group increased .Conclusion:Zinc sulfate alleviates the apoptosis of HaCaT cell induced by UVB because of the expression of MT .

The Role of Metallothioneins (MTs) in the Occurrence and Development of Hepatocellular Carcinoma and Mechanism Behind the Prevention and Treatment of Traditional Chinese Medicine / 世界科学技术-中医药现代化

It is a brief review of our research on the role of metallothioneins (MTs) in the occurrence and development of hepatocellular carcinoma (Hcc) in more than a decade,and the mechanism of prevention and treatment for Hcc in the field of TCM.It was found that MTs were required for the malignant proliferation of Hcc cells and their expressions significantly increased under different levels of stress.MTs may exert their effects on promoting proliferation via increasing ribosomal protein gene or other gene expressions.The over-expressions of MTs directly altered the balance of ions in the cell nucleus,and interacted on the genes promoting malignant proliferation.Hepatotoxicity and hepatic damage caused by DEN were reduced and relieved to different extents by the representative formula or constitute,which under the common therapeutic principle,by which MTs expressions were down-regulated as well.In conclusion,both effects above are important in the liver protection,inhibition of hyperplasia and carcinogenesis.

Changes and significance of metallothionein expression during hepatocarcinogenesis in C57BL/6J mice / 中国比较医学杂志

Objective To investigate the dynamic changes of metallothionein(MTs)gene expression and explore the important significance of MTs during hepatocarcinogenesis.Methods One hundred and twenty-five SPF 5 -8-week old male C57BL/6J mice were randomly divided into control group and model group.Diethylnitrosamine (DEN) was given to the mice at a dose of 100 mg/kg, ip, and 50 mg/kg, ip, in the first and next week, respectively.The mice were given ethanol (53%, 5 mL/kg/day, 5 days/week) from the third week of experiment till 35 weeks.At 1, 3, 9, 13, 24 and 35 weeks of the experiment, liver samples were taken for histopathological examination of liver damages and incidence of HCC. The liver index and malondialdehyde (MDA) of liver homogenate were determined.All liver tissue samples were examined by histopathology using hematoxylin and eosin (HE), Masson and reticular fiber staining.Real-time RT-PCR was used to analyze the mRNA expression level of liver metallothionein-1 /2 (MT-1 /2) in different periods.Results Progressive liver damages in model group mice were identified in different periods.Hepatocytes abnormal tission and abnormal liver plate structure, architecture often characteristic of HCC could be seen in approximately 50% of mice at 35 weeks.In addition to these, a higher liver index also were seen at 35 weeks.Increased MDA levels in the mouse liver tissues were observed in each stage.Real-time RT-PCR analysis showed that significantly increased transcription of MT-1 and MT-2 at 1, 3 and 9 weeks, then gradually declined and even below the normal level.Conclusions MTs gene expression levels in mouse liver tissues are changed from significantly increased in the early stage of injury to decreased expression combined with distinct fibrosis. Our findings further demonstrate that the down-regulation of MTs level is closely correlated with hepatocarcinogenesis.

Study on expression of metallothionein 2 gene in safflower / 中草药

Objective: To obtain the transgenic safflower plants which expressed Arabidopsis thaliana metallothionein 2 (MT2) gene, and lay a foundation for development of MT products. Methods: The oleosin-MT gene was obtained from pEASY-oleosin-MT by Nco I/Hind III, then was inserted into plant expression vector pOP. The recombinant plasmid named pOP-oleosin-MT was transferred into Agrobacterium tumefaciens EHA105.The oleosin-MT gene was introduced into safflowers via Agrobacterium-mediated method and positive transgenic plants were determined by PCR analysis. Results: The recombinant plasmid pOP-oleosin-MT was successfully constructed. PCR and Southern blotting analysis confirmed that MT gene was integrated into the genome of safflower plant and three transgenic plants were obtained. Conclusion: The safflower regeneration system is constructed successfully and MT gene is successfully transformed into safflower plant.

Cloning of MT-2 gene and its promoter in Pheretima aspergillum / 中草药

Objective: To lay a foundation for attenuating the heavy metal accumulation in Pheretima aspergillum by means of genetic engineering technology in further research, we revealed the transcriptional regulation mechanism of MT-2 gene. Methods: The coding sequence of MT-2 gene was amplified by PCR with specific primers, which were designed according to their known cDNA sequences, and the outcomes were contrastively analyzed after the sequencing process. Prior to the isolation of 5' promoter sequence by genome walking technology, three specific primers were designed based on MT-2 cDNA sequence. Meanwhile, the cis-acting elements of MT-2 gene were analyzed by Promoter Prediction online software. Results: After PCR and sequencing processes, a 2 826 bp coding sequence of MT-2 gene were obtained, four exons and four introns were found to compose the coding area by comparing with the known MT-2 cDNA sequence (accession No.KC787373.1). Besides, after genome walking and Promoter Prediction online analysing, a 1 534 bp promoter region of MT-2 was isolated, which contained not only CAAT box, TAAT box, and other core promoter elements, but also three MRE elements which specifically response to heavy metal involved in regulating the MT-2 expression. Conclusion: The expression of MT-2 gene in P. aspergillum can be induced by heavy metal, and the transcriptional level is achieved by MRE regulatory elements located in MT-2 gene promoter region.