Metformin Attenuates Cyclosporine A-induced Renal Fibrosis in Rats.

BACKGROUND: The aim of the present study was to investigate the therapeutic potential of metformin in preventing cyclosporine A (CsA)-induced nephrotoxicity. METHODS: Three groups of adult male Sprague-Dawley rats were treated with vehicle, CsA, and CsA + metformin for 4 weeks following 1 week on low sodium diet, respectively. At the end of treatment, all animals were euthanized, and the samples of kidney, urine, and blood were collected for functional, morphological, and molecular biological evaluation. RESULTS: Metformin effectively prevented CsA-induced renal dysfunction with increased creatinine clearance rate and reduced blood urea nitrogen and serum creatinine, as well as less proteinuria in comparison to the CsA group. Morphologically, metformin ameliorated CsA-induced renal fibrosis and tissue collapse in the areas of arteries, glomeruli, and proximal tubules. We further demonstrated that the antifibrotic effects of metformin in kidneys treated with CsA were associated with decreased phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2). CONCLUSIONS: In conclusion, our study revealed new therapeutic potential of metformin to attenuate calcineurin inhibitor-induced renal fibrosis, which was closely related to the suppression of MEK/ERK1/2 pathway.

MEK1 and MEK2 differentially regulate human insulin- and insulin glargine-induced human bladder cancer T24 cell proliferation.

BACKGROUND: Increased risk of bladder cancer has been reported in diabetic patients. This study was to investigate the roles of mitogen-activated protein kinase kinase (MEK) 1 and 2 in the regulation of human insulin- and insulin glargine-induced proliferation of human bladder cancer T24 cells. METHODS: In the absence or presence of a selective inhibitor for MEK1 (PD98059) or a specific siRNA for MEK2 (siMEK2), with or without addition of insulin or glargine, T24 cell proliferation was evaluated by cell counting kit (CCK)-8 assay. Protein expression of MEK2, phosphorylation of ERK1/2 and Akt was analyzed by Western blotting. RESULTS: T24 cell proliferation was promoted by PD98059 at 5 - 20 µmol/L, inhibited by siMEK2 at 25 - 100 nmol/L. PD98059 and siMEK2 remarkably reduced phosphorylated ERK1/2. Insulin- and glargine-induced T24 cell proliferation was enhanced by PD98059, suppressed while not blocked by siMEK2. Insulin- and glargine-induced ERK1/2 activation was blocked by PD98059 or siMEK2 treatment, whereas activation of Akt was not affected. CONCLUSION: MEK1 inhibits while MEK2 contributes to normal and human insulin- and insulin glargine-induced human bladder cancer T24 cell proliferation.

[Relationship of adverse pregnancy outcomes and a high risk serum screen for Down syndrome in the second trimester].

OBJECTIVE: To investigate the the relationship of a high risk serum screen for Down syndrome in second trimester and adverse pregnancy outcomes, and to evaluate the predictive value for adverse pregnancy outcomes. METHODS: The tri-marker second trimester maternal serum screening for Down syndrome (alpha-fetoprotein, free beta-hCG and unconjugated estriol) was performed on the pregnant women at Peking Union Medical Hospital from January 2009 to January 2011. The cutoff valvue was 1/270. Pregnancy outcomes were followed up. The general condition and pregnancy complications of the pregnant women with high risk (high-risk group) were compared to that of the pregnant women with low risk (low-risk group); and with 35 years old as a demarcation, the incidences of adverse pregnancy outcomes were calculated in the two groups. RESULTS: (1) A total of 1935 cases were collected. And 1784 cases were with low risk, and 151 cases were with high risk. The difference of weight and gestational age between the two groups was not statistically significant (P > 0.05); the difference of age between the two groups was statistically significant (P < 0.01). (2) Pregnancy complications were found in 791 cases. In high-risk group, the incidences of gestational diaetes mellitus (GDM, 13.9%), neonatal asphyxia (4.0%) and small for gestational age infant (SGA, 4.6%) were higher than that in low-risk group (8.4%, 1.0%, 1.6%), the difference was statistically significant (P < 0.05). The incidences of gestational hypertension disease, premature labor, oligohydramnios, placenta previa, placenta abruption, fetal macrosomia in the two groups was not statistically different (P > 0.05). (3) In 1705 cases aged less than 35 years, 129 cases (7.6%) were GDM, 43 cases (2.5%) were gestational hypertension disease, 61 cases (3.9%) were premature labor; in 230 cases aged 35 years or more, 41 cases (17.8%) were GDM, 12 cases (5.2%) were gestational hypertension disease, 15 cases (6.5%) were premature labor, and the difference between the two groups was statistically significant (P < 0.05). In < 35 years old group, the incidences of GDM, neonatal asphyxia and SGA (12.3%, 4.4%, 5.3%) were higher in the high-risk group than that (7.2%, 0.9%, 1.6%) in the low-risk group, and the difference was statistically significant (P < 0.05). In ≥ 35 years old group, the incidences of GDM, neonatal asphyxia and SGA (18.9%, 2.7%, 2.7%) were slightly higher in the high-risk group than that (17.6%, 1.6%, 1.6%) in the low-risk group, the difference between the two groups was not statistically significant (P > 0.05). CONCLUSIONS: The present study revealed apparent increase in the adverse pregnancy outcomes in women with a high risk of Down syndrome screening test. Advanced age is the most important risk factor for a high risk of Down syndrome screening test and adverse pregnancy outcomes. More attention should be attached to the patients whose age were < 35 years old and with a high risk of Down syndrome screening test.

Lipopolysaccharide-enhanced early proliferation of insulin secreting NIT-1 cell is associated with nuclear factor-kappaB- mediated inhibition of caspase 3 cleavage.

BACKGROUND: Increased levels of plasma lipopolysaccharide (LPS) have been found in obesity and diabetes patients. This study was to investigate the effect of LPS on pancreatic beta-cell viability and the involvement of caspase 3 in NIT-1 cell line. METHODS: Mouse insulinoma NIT-1 cells were treated with LPS for the indicated time and dose. Cell viability was measured by cell counting kit-8 reagent. Toll-like receptor 4 (TLR4), caspase 3 and cleaved caspase 3 were detected by Western blotting. Insulin was determined by radioimmunoassay (RIA). RESULTS: LPS promoted NIT-1 cell proliferation at 1 µg/ml, peaked at 72 hours of incubation. A reduction in cleavage of caspase 3 was observed upon LPS treatment. Bay11-7082, a specific inhibitor of nuclear factor (NF)-κB, blunted LPS-induced inhibition of caspase 3 cleavage. Reduction in chronic insulin secretion was observed after treatment with LPS at 1 µg/ml for 48 and 72 hours, not for 24 hours. TLR4 protein was upregulated when NIT-1 cells were treated with LPS at 1 µg/ml for 24 hours. CONCLUSIONS: LPS promotes early NIT-1 cell proliferation in association with NF-κB-mediated inhibition of caspase 3 cleavage. LPS exerts a time-dependent inhibitory effect on chronic insulin secretion from NIT-1 cells.

Cytogenetic and clinical analysis of 340 Chinese patients with primary amenorrhea.

OBJECTIVE: To analyze the relationship between karyotypes and clinic features of patients with primary amenorrhea. METHODS: G banding was done for 340 patients with primary amenorrhea to facilitate individual chromosome identification, and if specific staining for certain portions of the chromosome was necessary, C banding was used. The clinical data were recorded by physical examination and ultrasound scanning. RESULTS: Karyotype analysis of the 340 patients revealed that 180 (52.94%) patients had normal female karyotypes and 160 (47.06%) patients had abnormal karyotypes. The abnormal karyotypes included abnormal X chromosome (150 patients), mosaic X-Y chromosome (4 patients), abnormal autosome (5 patients), and X-autosome translocation (1 patient). The main clinical manifestations in patients with primary amenorrhea were primordial or absent uterus (95.9%), invisible secondary sex features (68.8%), little or absent ovary (62.6%), and short stature (30.0%). The incidence of short stature in patients with X chromosome aberration (46%, 69/150) was significangly higher that in patients with 46, XX (9.44%, 17/180) as well as 46, XY (6.67%, 3/45; Chi square = 146.25, P=0.000). All primary amenorrhea patients with deletion or break-point at Xp1 1.1-11.4 were short statures. CONCLUSIONS: One of the main reasons of primary amenorrhea is choromosome abnormality, especially heterosome abnormality. It implies the need to routinely screen chromosomal anomalies for such patients. There might be relationship between Xp1 1.1-11.4 integrity and height improvement.

[Correlation of a disintegrin and metalloprotease 12 level in maternal serum during the first trimester with pregnancy outcome].

OBJECTIVE: To explore the relationship between pregnant outcomes and the maternal serum level of a disintegrin and metalloprotease 12 (ADAM 12) in the first trimester. METHODS: From July 2007 to January 2008, the serum levels of ADAM 12 of 511 women in their first trimester (6 - 13 gestational weeks), who attended the clinics at Peking Union Medical College Hospital, were tested by Time-Resolved Fluorescence Immunoassay (TR-FIA), and the results and pregnant outcomes were analyzed. RESULTS: (1) The median levels of ADAM 12 at 6, 7, 8, 9, 10, 11, 12, 13 weeks of gestation were 14.63 microg/L, 35.08 microg/L, 88.90 microg/L, 186.51 microg/L, 370.62 microg/L, 537.71 microg/L, 632.55 microg/L, and 769.42 microg/L, respectively, showing a linear increase with the gestational age (r =0. 992, P < 0.01). (2) Among the 511 pregnancies, 427 were normal singleton term pregnancies and 84 had adverse perinatal outcomes. Twenty-seven miscarriages (5.3%, 27/511) and 5 ectopic pregnancies were reported and the Multiple of Medians (MOM) of them were 0.24 and 0.32, respectively, which was significantly lower than the normal singleton pregnancies (1.01, P < 0.05). However, the serum level of ADAM 12 in 5 women with placenta previa (MOM = 1.45) was significantly higher than the normal ones (P < 0.05). No significant correlation was found between the fetal birth weight and maternal serum level of ADAM 12 in the first trimester (r = -0.15, P < 0.05). (3) Thirteen cases with chromosomal abnormalities was identified out of 97 cases who received fetal karyotyping, including 3 Down's syndrome and 2 Turner syndrome, and the MOM of ADAM 12 in these 13 cases (0.34) was significantly different from those normal singleton pregnancies (P < 0.05). MoMs of ADAM 12 in 10 euchromosome aneupolyhaploids cases (0.29)were lower than the normal ones (P < 0.05). CONCLUSION: The maternal serum level of ADAM 12 in the first-trimester is a potential marker for aneupolyhaploid screening and early fetal loss prediction, and is suggested to be tested at 9-12 gestational weeks as part of prenatal screening.

[Second trimester maternal serum screening for Down's syndrome in mainland China: a multi-center prospective study].

OBJECTIVE: To evaluate the performance characteristics of the second trimester double-marker test for the detection of fetal Down's syndrome in mainland China. METHODS: This prospective national multi-centered study used alpha-fetoprotein (AFP) and free beta-subunit of human chorionic gonadotrophin (free beta-hCG) as the serum markers. From May 2004 to September 2006, 11 centers participated in the collection and analysis of maternal serum AFP and free beta-hCG between 14 and 20(+6) weeks of pregnancy. The screening results were calculated using the standard algorithm based on the standard database provided with the analytic software. Patients with an increased risk of Down's syndrome pregnancy (> or = 1/270) were offered genetic amniocentesis. Outcomes of all pregnancies were obtained. RESULTS: A total of 66 132 singleton pregnancies were included in the study. The median maternal age was 27 years. At a cut-off of 1 in 270, the detection rate (DR) based on a Caucasian database was 72% corresponding to a false positive rate (FPR) of 5%, and the DR based on the Chinese database was raised to 76% corresponding to an FPR of 5%. CONCLUSION: The double-marker test using AFP and free beta-hCG is an effective screen strategy for second-trimester detection of fetal Down's syndrome in mainland China. Ethnic variance exists between the Caucasian and Chinese populations. The accuracy of screening is increased by the use of race-specific medians.

[Effect of peroxisome proliferator-activated receptor-alpha agonist on adipokines expression in rats fed with high-fat diet].

OBJECTIVE: To explore the effect of peroxisome proliferator-activated receptor-alpha ( PPAR-alpha) agonist fenofibrate on adipokines expression in high-fat diet fed SD rats and its relationship to insulin resistance (IR). METHODS: Rats were randomized into three groups (n = 10) : HD group, fed with high-fat diet; HDF group, fed with high fat diet and treated with fenofibrate; and control group, fed with normal diet. Animals were sacrificed after 4-week follow-up. Plasma lipids, fasting plasma insulin, free fatty acids (FFA), and insulin sensitivity were detected. Reverse transcription-polymerase chain reaction was used to semi-quantitatively determine the mRNA expression of adipokines including tumor necrosis factor-alpha (TNF-alpha) , interleukin-6 (IL-6), angiotensinogen (AGT), angiotensin 11 type 1 receptor (AT1R), and adiponectin in brown fat. RESULTS: The plasma level of FFA, TG, and homeostatic model approach-IR index were (2. 37+/-0. 60) vs (1. 59+/-0. 30) vs (1. 33+/-0. 34 ) mmol/L, (0. 48+/-0. 11) vs (0. 30+/-0. 04) vs (0. 36+/-0. 07) mmol/L, and 12. 30+/-3. 97 vs 5. 03 +/-1. 88 vs 4. 17+/-1. 27 in the HD group, HDF group, and control group after 4 weeks of treatment with fenofibrate, respectively. The mRNA expressions of TNF-alpha and adiponectin were 1. 726+/-1. 408 vs 0. 713+/-0. 711 vs 0. 593+/-0. 382 and 0. 660+/-0. 192 vs 0. 949+/-0. 35 vs 0. 936+/-0. 130 in these three groups, which showed significant difference between HD group and HDF group (P < 0. 05 ) , while no significant difference between HDF group and control group (P > 0. 05). The mRNA expressions of AGT, AT1 R, and IL-6 had no significant difference among these three groups (P > 0. 05 ). CONCLUSION: PPAR-alpha agonist fenofibrate may reverse high-fat diet induced lipid abnormalities, improve insulin sensitivity, and regulate the mRNA expressions of TNF-alpha and adiponectin in adipose tissues.