Magnesium homeostasis is very important to normal functions of cell and plays a role in a number of diseases such as diabetes, hypertension, chronic respiratory disorder, osteoporosis and arrhythmia. MRS2 encode a magnesium transporter protein in Saccharomyces cerevisiae and deletion of MRS2 gene results in decrease in mitochondrial magnesium concentration, group Ⅱ RNA splicing defect and growth defect on nonfermentable carbon source. To obtain more information of magnesium homeostasis in mitochondria, mTn-lacZ/LEU2 transposon library was transformed into mrs2 deletion mutant to screen for suppressor genes of MRS2. YMR166C, a member of mitochondrial carrier family, was identified as a suppressor gene of MRS2. Deletion of YMR166C gene can rescue the defects of mrs2 deletion mutant such as the decrease in mitochondrial magnesium concentration, groupⅡ RNA splicing defect and growth defect on nonfermentable carbon source. For the first time it was demonstrated that YMR166C is involved in mitochondrial magnesium homeostasis.

MTM1 gene is essential for SOD2 activity and normal mitochondrial function. MTM1 deletion results in decreased SOD2 activity, impaired mitochondrial function and growth defect on nonfermentable carbon source. A yeast genomic library was transformed into mtm1 deletion mutant to screen for suppressor genes of MTM1. The damage caused by MTM1 deletion is irreversible and even overexpression of MTM1 can not rescue the growth defect of mtm1 deletion mutant. Another screening strategy was adopted: a plasmid overexpressing MTM1 was transformed into wild type before the MTM1 gene on chromosome was deleted. The resulting strain, designated YES2MTM1, was transformed with a yeast genomic library. Transformants lost the plasmid overexpressing MTM1 after 5-FOA treatment. Yeast strains able to grow on nonfermentable carbon source with MTM1 deletion and overexpression of some DNA fragments were picked up and candidate suppressor genes were identified. Overexpression of five genes were identified to be able to rescue the growth defect on nonfermentable carbon source. The study will provide reference for MTM1 gene function and screening for suppressor of genes whose deletion result in irreversible damage.

Objective To study the effects of quantitative 24-hour urinary protein on the thyroid hormone levels in patients with severe preeclamp-sia,and clarify the impact of severe urinary protein on hypothyroid in severe preeclampsia patients. Methods A total of 166 patients with severe pre-eclampsia were recruited for the study and divided into mild proteinuria group(2.0-4.9 g/d),midrange group(5-10 g/d)and severe group(>10 g/d)according to the quantitative 24-hour urinary protein. 268 healthy female individuals with normal blood pressure and uric routine in the same stage of pregnancy and of the same age were selected into control group. Serum thyrotropin(TSH),free triiodothyronine(FT3)and free thyroxine (FT4)levels were determined by solid-phase chemiluminescent enzyme immunoassay method(CMIA). The thyroid peroxidase antibody(TPOAb) and thyroglobulin antibody(TGAb)concentration were detected by electrochemiluminescent assay(ECLIA). Results TSH levels were signifi-cantly higher in patients comparing to the control group(P<0.01). In addition,severe group showed higher TSH levels than mild group(P<0.01). FT4 and FT3 levels were obviously decreased with the progression of the disease(P<0.01 and P<0.05). The positive rate of TPOAb in mild group was significantly higher than that in moderate group(OR=9.8,P<0.05). There was no significant difference of the TGAb positive rate among three patient groups(P>0.05). The incidence of subclinical hypothyroidism and clinical hypothyroidism in severe group was significantly higher than that in mild group and in control group(OR=2.5,P<0.05 and OR=9.0,P<0.05;OR=8.0,P<0.01 and OR=43.4,P<0.01). Conclusion Our re-sults indicated that 24-hour urine protein in severe preeclampsia patients has extensive effects on thyroid hormones levels. With the increasing of quantitative 24-hour urinary protein,the level of TSH increased and the FT4 decreased. Thyroid autoantibody positiveness has extensive effects on 24- hour urine protein. Incidence of hypothyroid increased with the increase of quantitative 24-hour urinary protein. 24-hour urinary protein quantitative was a risk factor for hypothyroidism in severe preeclampsia patients. More attention should be paid to the monitoring of 24-hour urinary protein in se-vere preeclampsia patients.

Objective To investigate the effects of probiotics on the occurrence of ovalbumin (OVA) induced food allergy and the regulatory effects on immune function of rat models. Methods Thirty female Brown-Norway rats aged 3 weeks were randomly divided into blank control group,food allergy group and probiotics intervention group(n=10).The levels of serum OVA-IgE and intestinal sIgA were measured by ELISA method.Splenic lymphocytes were isolated and cultured in vitro,and the Treg lymphocyte subgroups in the spleen were analyzed by flow eytometry.The levels of IFN-γ,IL-4 and IL-10 in the supernatant of cultured splenic lymphocytes were measured by ELISA method. Results The serum OVA-IgE level in food allergy group was significantly higher than that in blank control group(P<0.05),while that in probiotics intervention group was significantly lower than that in food allergy group(P<0.05).Compared with food allergy group and blank control group,the level of intestinal sIgA in probiotics intervention group was significantly higher(P<0.05).The percentage of CD4~+CD25~+T lymphoeytes in food allergy group was significantly lower than that in blank contml group(P0.05).The levels of IL-4 and IL-10 in food allergy group were significantly higher than those in blank control group(P<0.05).Probiotics intervention group had significantly lower levels of IL-4,IL-10 and IFN-γ than food allergy group(P<0.05).The ratio of IFN-γ/IL-4 in food allergy group was significantly lower than that in blank control group(P<0.05),while that in probiotics intervention group was significantly higher than that in food allergy group(P<0.05). Conclusion Probioties intervention could prevent the occurrence of food allergy in animal models by modulating the Th1/Th2 eytokine balance.

[ ABSTRACT] AIM: To investigate the effect of miR-496 over-expression on the growth and metastasis of colon cancer cells and its molecular mechanism .METHODS:The proteins interacting with miR-496 were screened by bioinfor-matic method.The levels of miR-496, CTNNB1 mRNA and β-catenin protein in colon cancer cell lines , HT29, HCT116 and SW480, and normal colonic epithelial cell line NCM 460 were detected by real-time PCR and Western blot .HT29, HCT116 and SW480 cells were transfected with miR-496 mimics using Lipofectamine 2000 and named as HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimics cells, respectively, and the cells transfected with the scramble served as negative control .The cell viability, lactate dehydrogenase (LDH) leakage, and colony formation and metastatic abilities were determined by MTT assay , LDH assay, colony formation assay and Transwell method , respective-ly.The promoter activity of miR-496 was measured using luciferase reporter gene assay .The protein levels of β-catenin, eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), p-4E-BP1, low-density lipoprotein receptor-related protein 6(LRP6), p-LRP6, MMP-7, MMP-9, MMP-13 and TIMP-2 were monitored by Western blot.RESULTS:Endog-enous miR-406 interacted with β-catenin was found in the colon cancer cells .Low miR-496 expression in the HT29, HCT116 and SW480 cells and high miR-496 expression in NCM460 cells were detected.In contrast, high β-catenin ex-pression was found in the HT29, HCT116 and SW480 cells and low β-catenin expression was observed in the NCM460 cells.Compared with control group , the cell viability, colony formation rate and the number of metastatic cells remarkably decreased in the HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimic cells (P<0.05).The promoter activity of miR-496 was significantly increased in colon cancer cells transfected with miR-496 mimics, and was 1.75, 2.04 and 1.61 times as high as control group .miR-496 over-expression inhibited β-catenin levels, and p-4E-BP1 and p-LRP6 protein levels were also reduced .siRNA-or over-expressed miR-496-mediated β-catenin down-regulation in-hibited MMP-7 and MMP-9 expression , but promoted TIMP-2 expression .CONCLUSION:The expression level of miR-496 in the colon cancer cells is low , but in the normal colonic epithelial cells is high .miR-496 over-expression inhibits the protein levels of MMP-7 and MMP-9, and promotes the protein expression of TIMP-2 via inhibiting Wnt/β-catenin path-way, thus suppressing malignant phenotype in the colon cancer cells .

Objective To study thyroid hormone changes in women with pre-eclampsia patients,the characteristics of thyroid disease and its relationship with pre-eclampsia.Methods From May 2011 to December 2012 171 patients with pre-eclampsia who delivered in Shengjing Hospital of China Medical University were recruited as prc-eclampsia(PE) group,among which 114 cases were defined as early onset pre-eclampsia (EP) group and 57 cases were defined as late onset pre-eclampsia (LP) group.And 171 healthy women with same age and same stage of pregnancy were selected as the control group.Their blood pressures were normal and they had no obstetrical complications.Serum thyrotropin (TSH),free triiodothyronine (FT3) and free thyroxine (FT4) levels were determined by solid-phase chemiluminescent enzyme immunoassay method (CMIA).Thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) were measured by electro-chemiluminescent assay (ECLIA).The positive rate was calculated (TPOAb ＞ 5.6 U/L,TGAb ＞ 4.1 U/L were defined as positive result).The relationship between TSH,FT3,FT4 level and blood pressure was analyzed in women with pre-eclampsia.Results (1) The median values of TSH,FT4 and FT3 in PE group were 3.4 mU/L,(12.0 ± 3.0) pmol/L and(3.9 ± 0.9) pmol/L.In the control group,they were 1.9 mU/L,(13.4 ± 2.4) and (5.0 ± 1.3) pmol/L.There were statistically significant differences between the two groups(P ＜ 0.01).In EP group,the median values of TSH,FT4 and FT3 were 3.3 mU/L,(12.1 ± 3.4) pmol/L and (3.8 ± 0.9) pmol/L.The differences between EP group and the control group were statistically significant (P ＜ 0.01).In LP group,the median values of TSH,FT4 and FT3 were 3.4 mU/L,(11.9 ± 3.1) pmol/L and (3.9 ± 1.0) pmol/L.There were statistically significant differences compared to the control group(P ＜0.01).While there was no difference between EP group and LP group (P ＞ 0.05).(2) The positive rate of TPOAb and TGAb in PE group were 15.2％ (26/171)and 21.6％ (37/171),and were 12.3％ (21/171) and 14.6％ (25/171) in the control group.There was statistically significant difference in the TGAb positive rate (P ＜ 0.01),but the difference in TPOAb positive rate was not statistically different(P ＞0.05).The TPOAb positive rates in EP group and LP group were 12.3 ％ (14/114) and 21.1％ (12/57),respectively,with no statistically significant difference (P ＞ 0.05).And the positive rates of TGAb in EP group and LP group were 21.9％ (25/114)and 21.1％ (12/57),respectively,with no statistically significant difference(P ＞ 0.05).The positive rate of TPOAb in LP group and in the control group had statistically significant difference(P ＜0.01).(3) The morbidity of thyroid disease in PE group and in the control group were 47.4％ (81/171) and 16.4％ (28/171),with statistically significant difference (P ＜ 0.01).(4) The morbidity of subclinical hypothyroidism or hypothyroidism in PE group and in the control group were 45.0％ (77/171) and 16.4％ (28/171),with statistically significant difference(P ＜0.01).(5) The morbidity of subclinical hyperthyroidism in PE group and in the control group were 2.3 ％ (4/171) and 1.8 ％ (3/171),with no statistically significant difference (P＞0.05).(6) In PE group,women with TSH level of 0.3-3.3 mU/L had systolic pressure of(170 ± 21)mmHg (1mmHg =0.133 kPa)and diastolic pressure of(112 ± 15) mmHg; women with TSH ＞ 3.3 mU/L had systolic pressure of(166 ± 21)mmHg and diastolic pressure of(109 ± 13)mmHg.There was no statistically significant difference(P ＞ 0.05).But the diastolic pressure in EP group and LP group had statistically significant difference(P ＜ 0.01).In PE group,no correlation was found among TSH,FT4 levels and systolic pressure,diastolic pressure(P ＞ 0.05).FT3 level was negatively correlated to diastolic pressure (r =-0.172,P =0.023).Conclusions It is common that pre-eclampsia is complicated with thyroid dysfunction,mainly subclinical hypothyroidism.Thus it is nessesary to test thyroid hormone and thyroid antibodies in women with pre-eclampsia.The decrease of FT3 and FT4,the increase of TSH and the presence of TPOAb and TGAb are related with the presence of pre-eclampsia.

To investigate the effects of component I from Agkistrodon acutus venom (AAVC-I) on the biological features of chronic myeloid leukemia cells, K562/A02 leukemia cells were cultured in the presence of AAVC-I (6.25 - 100 µg/ml) and the proliferation status was assayed by CCK-8 method. Morphological changes were observed by inversed microscope after Giemsa and Hochest 33258 staining, and cell apoptosis was detected by flow cytometry. Caspase 3 activity was tested by using Chromogenic Activity Assay Kit. The results showed that AAVC-I inhibited the growth of K562/A02 cells in time- and concentration-dependant manners, and the IC(50) at 48 h was 30.988 µg/ml. Giemsa and Hochest 33258 staining showed the typical apoptotic features in K562/A02 cells after induction with AAVC-I for 48 h. Flow cytometric analysis revealed that the percentage of the apoptotic cells reached from 0.88 up to 53.66 as the treated concentration was elevated from 0 to 50 µg/ml. Compared with the control group, the expression of caspase 3 in the tested group was enhanced in a dose-dependent manner (P < 0.05). It is concluded that AAVC-I can effectively inhibit the growth and promote apoptosis of K562/A02 cells. Elevated expression of caspase-3 may be attributed to the apoptosis of K562/A02 cells.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Crotalid Venoms ; pharmacology ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology

OBJECTIVETo explore the clinical characteristic of functional articulation disorders in children and provide more evidence for differential diagnosis and speech therapy.

METHODS172 children with functional articulation disorders were grouped by age. Children aged 4-5 years were assigned to one group, and those aged 6-10 years were to another group. Their phonological samples were collected and analyzed.

RESULTSIn the two groups, substitution and omission (deletion) were the mainly articulation errors in these children, dental consonants were the main wrong sounds, and bilabial and labio-dental were rarely wrong. In age 4-5 group, sequence according to the error frequency from the highest to lowest was dental, velar, lingual, apical, bilabial, and labio-dental. In age 6-10 group, the sequence was dental, lingual, apical, velar, bilabial, labio-dental. Lateral misarticulation and palatalized misarticulation occurred more often in age 6-10 group than age 4-5 group and were only found in lingual and dental consonants in two groups.

CONCLUSIONMisarticulation of functional articulation disorders mainly occurs in dental and rarely in bilabial and labio-dental. Substitution and omission are the most often occurred errors. Lateral misarticulation and palatalized misarticulation occur mainly in lingual and dental consonants.

Human xanthine oxidase is considered to be a target for therapy of hyperuricemia. Cichorium intybus is a Chinese plant medicine which widely used in Xinjiang against various diseases. In order to screen the inhibitors of xanthine oxidase from C. intybus and to explore main pharmacological actions of cichory a compound collection of C. intybus was built via consulting related references about chemical research on cichory. The three-dimensional crystal structure of xanthine oxidase (PDB code: 1N5X) from Protein Data Bank was downloaded.. Autodock 4.2 was employed to screen the inhibitors of xanthine oxidase from cichory 70 compounds were found to possess quite low binding free energy comparing with TEI (febuxostat). C. intybus contains constituents possessing potential inhibitive activity against xanthine oxidase. It can explain the main pharmacological actions of cichory which can significantly lower the level of serum uric acid.
Chicory ; chemistry ; Databases, Protein ; Drugs, Chinese Herbal ; chemistry ; Enzyme Inhibitors ; chemistry ; Humans ; Molecular Docking Simulation ; Molecular Structure ; Xanthine Oxidase ; antagonists & inhibitors ; metabolism

Objective To explore the value of 11C-choline PET/CT in patients with hepatic spaceoccupying lesions that have an indeterminate diagnosis by 18F-fluorodeoxyglucose (FDG) PET/CT. Methods A total of 25 liver masses in 20 patients with an indeterminate diagnosis based on 18F-FDG PET/CT were enrolled. Regional 11C-choline PET/CT scan was performed in all of the patients. Lesions with intense 11C-choline uptake were considered as positive. The semiquantitative maximum standardized uptake value(SUVmax) was measured and the tumor-to-liver (T/L) radioactivity ratio was calculated. The Mann-Whitney test,Kruskal-Wallis test and crosstabs x2-test were performed by using SPSS version 11.5. Results Of the 25 lesions,21 were proven to be hepatocellular carcinomas (HCC),3 hemangiomas,and 1 parasitic granuloma. The sensitivity of 11C-choline PET/CT for the detection of HCC was 66.7% (14/21). 11C-choline PET/CT had a higher sensitivity for well differentiated HCC than moderately and poorly differentiated HCC on a patient basis (8/9 vs 2/5,respectively). There were significant differences of 11C-choline T/L ratios between the HCC positive group,HCC negative group and benign lesion group ( 1.70 ± 0.35,0.86 ± 0.15,and 0.36 ± 0.18,x2 = 19.00,P ＜0.01 ). The lesion size and alphafetoprotein (AFP) level between the HCC positive and negative groups had no significant difference respectively ( Mann Whitney U = 39.00,P ＞0.05,and U=16.00,P＞0.05,respectively). Conclusions 11C-choline is complementary to 18F-FDG PET/CT for the detection of HCC,especially for well differentiated HCC.