GRP78 protein metabolism in obese and diabetic rats: a study of its role in metabolic disorders.

OBJECTIVE: This study aimed to compare and analyze the expression and significance of the GRP78 protein in cochlear cell injury induced by a high glucose and high-fat diet in obese and diabetic rats. METHODS: Male SD rats were randomly divided into two groups: normal (NC) and high-fat (HF) groups. The NC group was fed a standard diet for eight weeks, while the HF group received a high-glucose, high-fat diet. The HF group was further categorized into the obesity group (OB group) and the type II diabetes mellitus group (T2DM group). To induce a type II diabetes mellitus (T2DM) model, the T2DM group received an intraperitoneal injection of a small dose of STZ (45 mg/kg). After four weeks on the original diet, body weight, blood glucose, blood lipid levels, and auditory brainstem response (ABR) thresholds were measured. The cochlea was dissected, and its morphology was observed using HE staining. Immunohistochemistry and western blotting were utilized to examine the expression level of the GRP78 protein in the cochlea. RESULTS: (1) The ABR threshold demonstrated a statistically significant difference between the T2DM group and the OB group (P < 0.05), as well as between the OB group and the NC group (P < 0.05). (2) Based on morphological comparisons from HE-stained sections, the T2DM group exhibited the most significant alterations in the number of cells in the spiral ganglion, the organ of Corti, and the stria vascularis of the cochlea. (3) The expression level of the GRP78 protein in the cochlea was higher in the T2DM group compared to the OB group (P < 0.05) and higher in the OB group compared to the NC group (P < 0.05). CONCLUSION: The findings indicate that the GRP78 protein plays a role in hearing loss caused by T2DM and hyperlipidemia. Moreover, T2DM is more likely than hyperlipidemia to be associated with hearing impairment.

The relationship between patterns of insulin secretion and risks of gestational diabetes mellitus.

OBJECTIVE: To evaluate patterns of insulin secretion in pregnancy and analyze the association between insulin patterns and risks of gestational diabetes mellitus (GDM). METHODS: A prospective study was conducted to collect and analyze pregnant women's materials from January 2015 to December 2018. Pregnant women were grouped according to results of 75-g oral glucose tolerance test at 24-28 weeks of pregnancy: normal glucose tolerance (NGT) and GDM. Insulin secretion patterns were based on the time of peak(s) and shape of insulin secretion curve. The relationship between insulin secretion patterns and pregnant outcomes was analyzed. RESULTS: A total of 2432 pregnant women met the inclusion criteria during the study period. Among them, 737 (30.3%) women were grouped as GDM and 1695 (69.7%) as NGT. Type I insulin secretion represented the early phase of insulin secretion (peak time at 30 or 60 minutes), while type II represented the delayed peak of insulin secretion (peak time at 120 or 180 minutes). Logistic regression analysis showed that type II insulin secretion was a risk factor of pre-eclampsia, large-for-gestational-age, and neonatal hypoglycemia. CONCLUSION: The delayed insulin peak is a useful marker for risk of GDM and adverse pregnant outcomes in women with GDM.

Downregulated ABHD5 Aggravates Insulin Resistance of Trophoblast Cells During Gestational Diabetes Mellitus.

Lipid metabolism-associated molecule abhydrolase domain containing 5 (ABHD5) has been reported to have a role in insulin-mediated glucose uptake, the deregulation of it is associated with gestational diabetes mellitus (GDM). However, whether ABHD5 participates in glucose metabolism disorders in GDM patients has remained elusive. The present study aimed to clarify the role of ABHD5 in regulating insulin signaling in placentae during GDM. Reverse transcription-quantitative polymerase chain reaction (qRT-PCR) analysis was used for detecting the levels of ABHD5 and AMP kinase (AMPK), the insulin signaling molecules insulin receptor (INSR), INSR substrate (IRS1, IRS2), phosphoinositide 3-kinase (PI3K) and AKT, as well as the glucose transporter type 4 (GLUT-4) in placentae and the trophoblast cell line HTR-8/SVneo, while the protein level of ABHD5 was determined by western blotting. Pearson correlation analysis was performed to assess the correlation between the levels of ABHD5 and AMPK in placentae. In addition, ABHD5 overexpression in HTR-8/SVneo cells was achieved using plasmid vectors. The results indicated that the expression of ABHD5 and AMPK was dampened in placental tissues of females with GDM, and the levels of ABHD5 were positively correlated with AMPK. High-glucose (HG) treatment suppressed the expression of ABHD5, AMPK, GLUT-4, INSR, IRS, PI3K, and AKT in HTR-8/SVneo cells, and the overexpression of ABHD5 caused an elevation of the expression of these genes under normal and HG conditions in vitro. In conclusion, HG conditions induce insulin resistance of HTR-8/SVneo cells through downregulating ABHD5, which may account for impaired insulin signaling of placental tissues in GDM women.

MiR-137 Restricts the Viability and Migration of HTR-8/SVneo Cells by Downregulating FNDC5 in Gestational Diabetes Mellitus.

BACKGROUND: An increasing number of studies have described the pathological changes of placenta tissues in gestational diabetes mellitus (GDM), although the underlying mechanisms involved in this process remain uncertain. The aim of the present study was to verify the possible role of microRNA-137 (miR)-137 and FNDC5 in regulating the biological function of trophoblasts in high glucose (HG) conditions during the GDM period. METHODS: Expression levels of miR-137 and FNDC5 were measured in placenta specimens, the HG-treated trophoblast cell line HTR-8/SVneo and miR-137- overexpressing HTR-8/SVneo cells using reverse transcription quantitative-PCR or western blotting. The viability of HTR-8/SVneo cells was tested using a Cell Counting kit- 8 (CCK8) assay, with cell migration assessed using scratch and transwell assays. RESULTS: It was observed that the expression levels of miR-137 were increased and the expression levels of FNDC5 were decreased in the placenta tissues of women with severe GDM and in HG-exposed HTR-8/SVneo cells. In addition, upregulating miR-137 in HTR-8/SVneo cells downregulated the expression levels of FNDC5. The viability and migration of HTR-8/SVneo cells were suppressed by increased miR-137 expression levels, and upregulating FNDC5 in miR-137-overexpressing HTR-8/SVneo cells resulted in the reversal of all these effects. CONCLUSIONS: The data from the present study suggest that miR-137 suppresses the viability and migration of trophoblasts via downregulating FNDC5 in GDM, which may contribute to the pathology of placenta tissues and occurrence of adverse pregnancy outcomes.

High glucose suppresses the viability and proliferation of HTR­8/SVneo cells through regulation of the miR­137/PRKAA1/IL­6 axis.

The aim of the present study was to investigate the mechanism underlying the high glucose (HG)­associated regulation of HTR­8/SVneo cell viability and proliferation during gestational diabetes mellitus (GDM), and to verify the association of microRNA (miR)­137, protein kinase AMP­activated catalytic subunit α1 (PRKAA1) and interlukin­6 (IL­6). miR­137­overexpressing and negative control HTR­8/SVneo cells were established by lentiviral vector infection. Cell Counting Kit­8 and colony formation assays were used to analyze the viability and proliferation of HTR­8/SVneo cells. Reverse transcription­quantitative polymerase chain reaction analysis was used to determine the transcriptional activity of miR­137, PRKAA1 and Il­6, and ELISA and western blot analysis were used to measure the protein levels of IL­6 and PRKAA1, respectively. It was demonstrated that PRKAA1 was decreased in the placental tissues of women with GDM and HG­treated HTR­8/SVneo cells, and that HG upregulated miR­137 and IL­6 in trophoblasts. The overexpression of miR­137 decreased levels of PRKAA1 and increased levels of IL­6 in the HTR­8/SVneo cells. An inhibitor of PRKAA1 promoted the secretion of IL­6, whereas an agonist of PRKAA1 suppressed the production of IL­6. HG treatment and the overexpression of miR­137 reduced the viability and proliferation of HTR­8/SVneo cells in vitro, whereas the activation of PRKAA1 or incubation with IL­6 antibody reversed these effects. Overall, it was concluded that HG suppressed the viability and proliferation of trophoblast cells through the miR­137/PRKAA1/IL­6 axis, which may contribute to pathological changes of placental tissues in GDM.

High glucose induces dysfunction of human umbilical vein endothelial cells by upregulating miR-137 in gestational diabetes mellitus.

Recent studies have revealed considerable dysfunction of vascular endothelial cells (VECs) and abnormal expression of microRNA (miR)-137 in women with gestational diabetes mellitus (GDM), and the aim of this study was to clarify the underlying mechanism and possible role of microRNA (miR)-137 in dysfunction of VECs during GDM. We found increased levels of miR-137 in the plasma of GDM women and high-glucose (HG)-exposed HUVECs. Upregulating miR-137 in HUVECs elevated the chemokine (C-C motif) ligand 2 (CCL2) secretion and enhanced the chemotaxis and adhesion of U937 and THP-1 (two human acute monocytic leukemia cell lines) cells to HUVECs in a co-culture system. Moreover, HG stimulation and/or overexpression of miR-137 inhibited the viability, upregulated the expression levels of vascular cell adhesion molecule-1 (VCAM-1), intercellular cell adhesion molecule-1 (ICAM-1), E-selectin, and inflammatory cytokine interleukin (IL)-6, and downregulated the production of IL-8, vascular endothelial growth factor (VEGF), and angiogenesis of HUVECs in vitro. These results imply that up-regulated miR-137 by HG can restrict the viability and angiogenesis, promote the activation and inflammatory cytokine secretion of VECs, and stimulate the monocyte chemotaxis and adhesion to VECs. Ultimately, we have concluded that miR-137 is crucial to HG-induced VEC dysfunction and may be involved in pathology of GDM.

Emended descriptions of the species Sphingomonas adhaesiva Yabuuchi et al. 1990 and Sphingomonas ginsenosidimutans Choi et al. 2011.

During the phylogenetic study of the genus Sphingomonas and its closely related genera, we found that there existed errors in the 16S rRNA gene sequence of the type strain of the type species of Sphingomonas adhaesiva (D13722). Data suggested the wrong sequence should be replaced by the sequence under the accession number KY927401. As the new sequence shared 99.6 % 16S rRNA gene sequence similarity with that of Sphingomonas ginsenosidimutans, the relationship between these two species was reevaluated in the present study. Analyses, based on the whole genome sequences, phenotypic characteristics and fatty acid profiles clearly show that S. adhaesiva and Sphingomonas ginsenosidimutans are two distinct species of the genus Sphingomonas. Considering the errors in the original descriptions of S. adhaesiva and S. ginsenosidimutans, we have emended the descriptions of the two species.

Genome-based reclassification of Sphingopyxis ummariensis as a later heterotypic synonym of Sphingopyxis terrae, with the descriptions of Sphingopyxis terrae subsp. terrae subsp. nov. and Sphingopyxis terrae subsp. ummariensis subsp. nov.

During the phylogenetic analysis of the genus Sphingopyxis, we found that an incorrect 16S rRNA gene sequence (accession number: D13727) was provided in the original description of Sphingopyxisterrae NBRC 15098T and the wrong sequence has been adopted and used for a long time. It should be replaced by the new correct 16S rRNA gene sequence (accession number: MF618306). The new sequence shared the highest similarity (99.8 %) with that of Sphingopyxisummariensis DSM 24316T. The average nucleotide identity (ANI) (96.87 %) and digital DNA-DNA hybridization (75.30 %) values based on the whole-genome sequences and almost the same phenotypic and chemotaxonomic characteristics of the two type strains revealed thatSphingopyxisummariensis should be a later heterotypic synonym ofSphingopyxisterrae. However, the distinctions in the genome size, hydrolysis of aesculin, α-glucosidase and particularly the fatty acid profiles strongly support that strain DSM 24316T should be considered to represent a novel subspecies ofSphingopyxisterrae. Two novel subspecies are therefore proposed, namely Sphingopyxisterraesubsp. terrae subsp. nov. (type strain E-1-AT=NBRC 15098T=JCM 10195T=DSM 8831T=LMG 17326T) and Sphingopyxisterraesubsp. ummariensis subsp. nov. (type strain UI2T=DSM 24316T=CCM 7428T=MTCC 8591T).

Sphingomonas difficilis sp. nov., a difficultly cultivable bacterium that grows on solid but not in liquid medium, isolated from an abandoned lead-zinc mine.

A difficult to cultivate bacterial strain, designated 1PNM-26T, isolated from a lead-zinc mine, was investigated using a polyphasic taxonomic approach. The strain was able to grow on solid medium but not in liquid medium. Cells were Gram-reaction-negative, aerobic, non-spore-forming, non-motile and rod-shaped. It showed positive reactions for catalase and oxidase and hydrolysis of aesculin. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that strain 1PNM-26T represents a member of the genus Sphingomonas and forms a stable cluster with Sphingomonas morindae KCTC 42183T, Sphingomonas polyaromaticivorans JCM 16711T and Sphingomonas oligoaromativorans NBRC 105508T. The major fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0. C14 : 0 2-OH was present as the major hydroxyl fatty acid. The major polyamine was sym-homospermidine, and ubiquinone 10 (Q-10) was the predominant respiratory quinone. The genomic DNA G+C content of strain 1PNM-26T was determined to be 66.3±0.3 mol%, and the polar lipids consisted of sphingoglycolipid, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidyldimethylethanolamine, phosphatidylmonomethylethanolamine, an unidentified glycolipid, three unidentified aminolipids and three unidentified lipids. The phenotypic, phylogenetic and chemotaxonomic results strongly supported the hypothesis that strain 1PNM-26T represents a novel species of the genus Sphingomonas, for which the name Sphingomonasdifficilis sp. nov. is proposed. The type strain is 1PNM-26T (=GDMCC 1.664T=KCTC 42758T=DSM 27573T).

Reclassification of Sphingopyxis contaminans as Sphingorhabdus contaminans comb. nov. and emended description of the genus Sphingorhabdus.

With the description of the genus Sphingorhabdus, the taxonomic position of Sphingopyxis contaminans was re-evaluated based on analyses of 16S rRNA gene sequences and phenotypic and chemotaxonomic characteristics. The results revealed that Sphingopyxis contaminans is clearly a member of the genus Sphingorhabdus and we proposed that Sphingopyxis contaminans(Subhash Y, Sasikala C, Ramana CV. Int J Syst Evol Microbiol 2014;64:2238-2243) should be reclassified as Sphingorhabduscontaminans comb. nov. An emended description of the genus Sphingorhabdus is also provided.

Sphingomonas spermidinifaciens sp. nov., a novel bacterium containing spermidine as the major polyamine, isolated from an abandoned lead-zinc mine and emended descriptions of the genus Sphingomonas and the species Sphingomonas yantingensis and Sphingomonas japonica.

A yellow-pigmented bacterial strain, designated 9NM-10T, was isolated from an abandoned lead-zinc mine in Meizhou, Guangdong Province, China. Cells were strictly aerobic, Gram-stain-negative and motile with a polar monotrichous flagellum. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain 9NM-10T belongs to the genus Sphingomonas and was most closely related to Sphingomonas yantingensis JCM 19201T and Sphingomonas japonica JCM 15438T. DNA-DNA relatedness values between strain 9NM-10T and these two type strains were 43.6±1.3 and 35.4±0.9 %, respectively. It contained Q-10 as the predominant respiratory quinone and the major cellular fatty acids were C18 : 1ω7c, C16 : 0, C17 : 1ω6c and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The genomic DNA G+C content of strain 9NM-10T was 68.7±0.2 mol%. The polar lipids were sphingoglycolipid, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, diphosphatidylglycerol, an unidentified phospholipid and three unidentified lipids. Strain 9NM-10T contained spermidine as the major polyamine. On the basis of phenotypic, phylogenetic and chemotaxonomic analyses, strain 9NM-10T is considered to represent a novel species of the genus Sphingomonas, for which the name Sphingomonas spermidinifaciens sp. nov. is proposed. The type strain is 9NM-10T (=GDMCC 1.657T=DSM 27571T). Descriptions of the genus of Sphingomonas and the species Sphingomonas yantingensis and Sphingomonas japonica were also emended in this study.

Genetic diversity of banana bunchy top virus isolates from China.

Banana bunchy top virus (BBTV) (the genus Babuvirus, the family Nanoviridae) is a single-stranded circular DNA virus with a genome composed of six components designated as DNA-R, -U3, -S, -M, -C, and -N. This study analyzed the nucleotide identities of the DNA-R of 23 isolates from banana-producing provinces of China, including Guangdong, Hainan, Guangxi, and Yunnan. Results showed that the nucleotide identity of DNA-R was 72.3-100%. Phylogenetic analysis indicated that these BBTV isolates were clustered in different subgroups within the Asian group (AG). Sequence analysis of the five other components (DNA -U3, -S, -M, -C, and -N) of the five isolates from China confirmed the results established for DNA-R of these BBTV isolates. This study suggested that the variation of DNA-R from Chinese BBTV isolates was considerably higher than the variation of other AG isolates, but their genetic diversity was low.

Sphingomonas metalli sp. nov., isolated from an abandoned lead-zinc mine.

A Gram-stain-negative and non-motile bacterial strain designated 9O-5T was isolated from an abandoned lead-zinc mine in Meizhou, Guangdong Province, southern China. The isolate was orange-pigmented, aerobic, and oxidase- and catalase-positive. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain 9O-5T belongs to the genus Sphingomonas and was closely related to Sphingomonas abaci DSM 15867T (97.6 % similarity), Sphingomonas phyllosphaerae FA2T (96.9 %) and Shingomonas guangdongensis 9NM-8T (96.8 %). Mean DNA-DNA relatedness between strain 9O-5T and S. abaci DSM 15867T was only 47.1 ± 4.9 %. The major fatty acids were C18 : 1ω7c, C14 : 0 2-OH and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). It contained Q-10 as the predominant respiratory quinone and sym-homospermidine as the major polyamine. The polar lipids were sphingoglycolipid, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, five unidentified phospholipids and six unidentified lipids. The genomic DNA G+C content of strain 9O-5T was 69.1 ± 0.1 mol%. Based on the data from this polyphasic taxonomic study, strain 9O-5T should be considered as representing a novel species of the genus Sphingomonas, for which the name Sphingomonas metalli sp. nov. is proposed. The type strain is 9O-5T ( = CGMCC 1.15330T = KCTC 42759T).

Association between glycemic control and birthweight with glycated albumin in Chinese women with gestational diabetes mellitus.

AIMS/INTRODUCTION: To assess glycated albumin (GA) as a potential glycemic index in managing gestational diabetes mellitus (GDM). MATERIALS AND METHODS: Eligible pregnant women were divided into the GDM group with abnormal result on a 75-g oral glucose tolerance test (OGTT) and the control (normal) group. GA measurements, Pearson's correlation analysis, multiple logistic regression and receiver operating characteristic curve analysis were obtained at the follow-up examination of participants in the two groups. RESULTS: A total of 2,118 women were assigned to the GDM group (n = 639) and control group (n = 1,479). The mean level of serum GA in GDM group was significantly greater than that in the control group at both 24-28 and 36-38 weeks of gestation (P < 0.05). The area under the receiver operating characteristic curve for GA defining good glycemic control in GDM was 0.874 (95% confidence interval 0.811-0.938). The cut-off point for the GA levels derived from the receiver operating characteristic curve was 11.60%, which had sensitivity and specificity for detecting a poor glycemic status of 75.93% and 86.36%, respectively. The risk of birthweight ≥3,500 g and macrosomia increased significantly with GA levels ≥13.00% at 24-28 weeks and ≥12.00% at 36-38 weeks of gestation. CONCLUSIONS: GA might be an appropriate and conveniently measured index that can detect poor glycemic control and predict birthweights in GDM women.

Massilia putida sp. nov., a dimethyl disulfide-producing bacterium isolated from wolfram mine tailing.

A heavy metal-resistant and dimethyl disulfide-producing bacterial strain, designated 6NM-7T, was isolated from wolfram mine tailing, Dayu County, Jiangxi Province, PR China. Strain 6NM-7T was aerobic, Gram-stain-negative and motile by means of a single polar flagellum. Phylogenetic analysis, based on 16S rRNA gene sequences, showed that strain 6NM-7T was affiliated with the genus Massilia and was closely related to Massilia norwichensis LMG 28164T (98.8 % 16S rRNA gene sequence similarity), Massilia kyonggiensis KACC 17471T (98.4 %), Massilia niastensis KACC 12599T (97.8 %), Massilia tieshanensis KACC 14940T (97.3 %), Massilia haematophila KACC 13771T (97.2 %), Massilia namucuonensis CGMCC 1.11014T (97.1 %) and Massilia aerilata KACC 12505T (97.1 %). The DNA-DNA relatedness values between strain 6NM-7T and its closely related type strains were all below 70 %. The major respiratory quinone was unbiquinone 8 (Q-8) and the major cellular fatty acids consisted of C16 : 0 (33.2 %), summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH; 21.8 %), C17 : 0 cyclo (20.8 %), C18 : 1ω7c (7.4 %) and C10 : 0 3-OH (5.8 %). The major polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The genomic DNA G+C content of strain 6NM-7T was 66.8 ± 0.6 mol%. On the basis of the results of this polyphasic taxonomic study, strain 6NM-7T should be assigned to a novel species of the genus Massilia, for which the name Massilia putida sp. nov. is proposed. The type strain is 6NM-7T ( = DSM 27523T = KCTC 42761T).

Transcriptome and expression profile analysis of highly resistant and susceptible banana roots challenged with Fusarium oxysporum f. sp. cubense tropical race 4.

Banana wilt disease, caused by the fungal pathogen Fusarium oxysporum f. sp. cubense 4 (Foc4), is regarded as one of the most devastating diseases worldwide. Cavendish cultivar 'Yueyoukang 1' was shown to have significantly lower disease severity and incidence compared with susceptible cultivar 'Brazilian' in greenhouse and field trials. De novo sequencing technology was previously performed to investigate defense mechanism in middle resistant 'Nongke No 1' banana, but not in highly resistant cultivar 'Yueyoukang 1'. To gain more insights into the resistance mechanism in banana against Foc4, Illumina Solexa sequencing technology was utilized to perform transcriptome sequencing of 'Yueyoukang 1' and 'Brazilian' and characterize gene expression profile changes in the both two cultivars at days 0.5, 1, 3, 5 and 10 after infection with Foc4. The results showed that more massive transcriptional reprogramming occurs due to Foc4 treatment in 'Yueyoukang 1' than 'Brazilian', especially at the first three time points, which suggested that 'Yueyoukang 1' had much faster defense response against Foc4 infection than 'Brazilian'. Expression patterns of genes involved in 'Plant-pathogen interaction' and 'Plant hormone signal transduction' pathways were analyzed and compared between the two cultivars. Defense genes associated with CEBiP, BAK1, NB-LRR proteins, PR proteins, transcription factor and cell wall lignification were expressed stronger in 'Yueyoukang 1' than 'Brazilian', indicating that these genes play important roles in banana against Foc4 infection. However, genes related to hypersensitive reaction (HR) and senescence were up-regulated in 'Brazilian' but down-regulated in 'Yueyoukang 1', which suggested that HR and senescence may contribute to Foc4 infection. In addition, the resistance mechanism in highly resistant 'Yueyoukang 1' was found to differ from that in middle resistant 'Nongke No 1' banana. These results explain the resistance in the highly resistant cultivar and provide more insights in understanding the compatible and incompatible interactions between banana and Foc4.

Butyrate alleviates metabolic impairments and protects pancreatic ß cell function in pregnant mice with obesity.

The relative or absolute deficiency of pancreatic ß-cell mass function underlies the pathogenesis of diabetes. It is necessary to alleviate the metabolic stress and reduce the demand for insulin to decrease the effects of mutations affecting ß-cell expansion. Butyrate is a natural nutrient existed in food and can also be produced physiologically through the intestinal fermentation of fiber. Pregnancy and obesity model would be helpful for understanding how ß-cell adapt to insulin resistance and how butyrate alleviate the metabolic impairment and protect pancreatic ß cell function in pregnant mice with obesity. C57BL/6J female mice were divided into three groups and fed with high fat food (HF group, 40% energy from fat), high fat with sodium butyrate food (HSF group, 95% HF with 5% butyrate), or control food (CF group, 14% energy from fat), respectively. The feeding would last for 14 weeks before mating and throughout the gestation period. A subset of dams were sacrificed at gestational day (GD) 14.5 to evaluate the changes of metabolism and ß-cell function, mass, proliferation and apoptosis, inflammatory reaction of islet from different diet. Pancreases were double immuno-labeled to assess the islet morphology, insulin expression, expression of proliferation gene PCNA and anti-apoptosis gene bcl-2. Moreover, we detected the expression of NF-κB, phosphorylated NF-κB (pNF-κB) to evaluate the islet inflammatory response with immunohistochemistry. Mice fed with HSF showed obviously changes including the decreased values of weight gain, glucose, insulin, triglyceride and total cholesterol level of blood compared with high fat diet group, and the reduced circulating maternal pro-inflammation factors at GD14.5. Mice fed with HF displayed ß-cell hyperplasia with a greater ß-cell size and ß-cell area in pancreas. Furthermore, the higher ratio of apoptosis and inflammatory response were found in HF group compared with HSF and CF group, while the proliferation rates of ß-cell increased in HF group, but not in HSF or CF. Butyrate shows an obvious function of anti-obesity, and can alleviate the metabolic stress, maintain the ß-cell function and protect them from inflammatory response in pregnant obese mouse without obvious fetus toxicity.

Gestational diabetes induces chronic hypoxia stress and excessive inflammatory response in murine placenta.

Metabolic impairments in maternal obesity and gestational diabetes mellitus (GDM) induce an abnormal environment in peripheral blood and cause vascular structure alterations which affect the placental development and function. A GDM model was developed using C57BL/6J female mice fed with high fat food (HF) (40% energy from fat) and a control group with control food (CF) (14% energy from fat) for 14 weeks before mating and throughout the gestation period. A subset of dams was sacrificed at gestational day (GD) 18.5 to evaluate the fetal and placental development. HF-fed dams exhibited significant increase in the maternal weight gain and homeostasis model assessment for insulin resistance index (HOMA-IR), impaired insulin secretion of glucose stimulus and glucose clearance of insulin stimulus before pregnancy; in addition, they also had the increase in the fetal and placental weight. HF-fed dams at GD 18.5 showed the high level of circulating maternal inflammation factors and were associated with increased oxidative stress and hypoxia in the labyrinth, abnormal vascular development with a high level of hypoxia inducible factor-1α (HIF-1α) and VEGF-A expression, but without a parallel increase in CD31 level; were induced an exaggerated inflammatory response in placental vascular endothelial cell. Our findings show that GDM induces more maternal weight gain and fetus weight, with abnormal maternal circulating metabolic and inflammation factors, and forms a placental hypoxia environment and impacts the placental vascular development. Our findings indicate that gestational diabetes induce excessive chronic hypoxia stress and inflammatory response in placentas which may contribute mechanisms to the high risks of perinatal complications of obesity and GDM mothers.

RNA silencing suppressor Pns11 of rice gall dwarf virus induces virus-like symptoms in transgenic rice.

Transgenic rice (Oryza sativa) plants expressing the Pns11 protein of rice gall dwarf virus (RGDV) displayed multiple abnormal phenotypes, some of which were highly reminiscent of the symptoms observed in RGDV-infected rice. Further analysis indicated that the apparent alterations in plant growth and morphology were correlated with the expression levels of microRNA160, microRNA162, microRNA167, microRNA168, and the microRNA target OsARF8. Especially, the striking dwarfing phenotype depended on the high expression level of microRNA167. By analogy to other categories of plant viruses, the RNA silencing suppressors encoded by plant dsRNA viruses function as pathogenicity determinants. These findings significantly deepen our current mechanistic understanding of the RNA silencing suppressor (VSR) encoded by a dsRNA virus and provide additional evidence that interference with microRNA expression is a VSR function utilized by a diverse range of viruses.

Chemokine CCL2 enhances survival and invasiveness of endometrial stromal cells in an autocrine manner by activating Akt and MAPK/Erk1/2 signal pathway.

OBJECTIVE: To clarify the role and mechanism of CCL2 in regulating the biological functions of endometrial stromal cells (ESCs). DESIGN: The CCL2 effect on the viability, proliferation, and invasion in the eutopic ESCs from endometriosis. SETTING: Research laboratories. PATIENT(S): Patients with endometriosis aged 23-47 years. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Signal transduction and downstream molecules from CCR2. RESULT(S): We have found that the secretion of CCL2 by the eutopic ESCs from endometriosis is higher than that of healthy ESCs without endometriosis. The CCL2 can enhance the viability, proliferation, and invasion of ESCs in a dosage and time-dependent manner. Anti-CCL2 neutralizing antibody and CCR2 antagonist can completely abolish the increase in viability, proliferation, and invasiveness of ESCs induced by CCL2. The CCL2 can increase the expression of proliferating cell nuclear antigen, survivin, and matrix metalloproteinase 2, and decrease the expression of tissue inhibitor of metalloproteinase 1 and 2, and promote the viability, proliferation and invasiveness of ESCs by activating Akt and MAPK/Erk1/2 signal pathway, but not p38 and JNK signal pathway. CONCLUSION(S): CCL2 might play an important role in regulating the functions of ESCs through Akt and MAPK/Erk1/2 signal pathway, and overexpression of CCL2 in ESCs and peritoneal fluid (PF) would lead to onset and development of endometriosis.