Serum homocysteine concentration as a marker for advanced diabetic nephropathy in a cohort of elderly patients.

BACKGROUND: Hyperhomocysteinemia has been linked with chronic kidney disease (CKD). The present study investigated whether homocysteine (Hcy) serum levels might serve as a marker for the advancement of diabetic nephropathy (DN). METHODS: Clinical and laboratory indicators including Hcy, vitamin D (VD), urine protein, estimated glomerular filtration rate (eGFR) and the urinary protein/creatinine ratio in subjects > 65 years with DN (n = 1,845), prediabetes (n = 1,180) and in a non-diabetes (control) group (n = 28,720) were analyzed. RESULTS: DN patients had elevated Hcy concentrations, decreased VD and higher urinary protein levels, a reduced eGFR and a higher urinary protein/creatinine ratio compared with prediabetic and control subjects. After correcting for urinary protein quantitation, multivariate analysis revealed that both the Hcy concentration (P < 0.010) and urinary protein/creatinine ratio (P < 0.001) were risk factors, while the VD2 + VD3 serum concentration (P < 0.001) was a protective factor for DN. Moreover, Hcy > 12 µmol/L was a cut-off value for predicting advanced DN. CONCLUSION: Hcy serum concentration is a potential marker for the advancement of CKD in DN but not prediabetes patients.

Proteomic profiles and the function of RBP4 in endometrium during embryo implantation phases in pigs.

BACKGROUND: Endometrial receptivity plays a vital role in the success of embryo implantation. However, the temporal proteomic profile of porcine endometrium during embryo implantation is still unclear. RESULTS: In this study, the abundance of proteins in endometrium on days 9, 10, 11, 12, 13, 14, 15 and 18 of pregnancy (D9, 10, 11, 12, 13, 14, 15 and 18) was profiled via iTRAQ technology. The results showed that 25, 55, 103, 91, 100, 120, 149 proteins were up-regulated, and 24, 70, 169, 159, 164, 161, 198 proteins were down-regulated in porcine endometrium on D10, 11, 12, 13, 14, 15 and 18 compared with that on D9, respectively. Among these differentially abundance proteins (DAPs), Multiple Reaction Monitoring (MRM) results indicated that S100A9, S100A12, HRG and IFI6 were differentially abundance in endometrial during embryo implantation period. Bioinformatics analysis showed that the proteins differentially expressed in the 7 comparisons were involved in important processes and pathways related to immunization, endometrial remodeling, which have a vital effect on embryonic implantation. CONCLUSION: Our results reveal that retinol binding protein 4 (RBP4) could regulate the cell proliferation, migration and apoptosis of endometrial epithelial cells and endometrial stromal cells to affect embryo implantation. This research also provides resources for studies of proteins in endometrium during early pregnancy.

Case report of pyogenic meningitis after tooth extraction in an elderly diabetic patient.

INTRODUCTION: Among elderly diabetic patients with comorbid conditions, the clinical characteristics of suppurative meningitis may not be typical, which is easy to cause misdiagnosis and delayed treatment. CASE PRESENTATION: In this report, we described the case of a 65-year-old elderly diabetic male who developed purulent meningitis after tooth extraction. The patient developed repeated infections (fever, headache and other symptoms) and was diagnosed with pyogenic meningitis by combining clinical manifestations and cerebrospinal fluid testing. The patient was treated with intracranial pressure reduction, nutritional support and anti-infection drugs, and recovered and was discharged after 15 days. CONCLUSIONS: The atypical clinical presentation and insidious onset of the disease in this patient could easily lead to missed diagnosis and misdiagnosis. We emphasize that we should be vigilant with purulent meningitis and active treatment and prevention should be implemented in this subgroup of patients. During treatment, active anti-infection and nutritional support should be provided on the basis of blood glucose control.

Extracellular vesicles derived from endometrial epithelial cells deliver exogenous miR-92b-3p to affect the function of embryonic trophoblast cells via targeting TSC1 and DKK3.

BACKGROUND: Extracellular vesicles (EVs) could mediate embryo-maternal communication to affect embryo implantation by delivering biology information, including microRNA (miRNA), protein, lipid. Our previous research shows that miR-92b-3p was differentially expressed in EVs of uterine flushing fluids during the embryo implantation period. However, the role of miR-92b-3p from EVs in embryo implantation remains elusive. MATERIALS AND METHODS: EVs were isolated from porcine endometrial epithelial cells (EECs) by ultracentrifugation. MiR-92b-3p mimics and EVs were used to regulate the expression of miR-92b-3p in porcine trophoblast cells (PTr2 cells). Cell proliferation, migration and adhesion analyses were used to observe the phenotype. RT-qPCR, western blot and dual-luciferase reporter assay were used to assess the targets of miR-92b-3p. RESULTS: In this study, EVs derived from porcine EECs were identified and could be taken up by PTr2 cells. We found that the EVs derived from EECs transfected with miR-92b-3p mimic (EVs-miR-92b-3p) significantly promoted the proliferation, migration and adhesion of PTr2 cells. We verified that Tuberous sclerosis complex subunit (TSC1) and Dickkopf 3 (DKK3) were the target genes of miR-92b-3p. Moreover, our study showed that miR-92b-3p plays a vital role in PTr2 cells via targeting TSC1 and DKK3. Furthermore, the 3'UTR vectors of TSC1 and DKK3 can rescue the effect of miR-92b-3p on PTr2 cells. CONCLUSIONS: Taken together, this study reveals a novel mechanism that EVs derived from porcine EECs treated with miR-92b-3p crosstalk with trophoblasts by targeting TSC1 and DKK3, leading to an enhanced ability for implantation.

Transcriptome regulation of extracellular vesicles derived from porcine uterine flushing fluids during peri-implantation on endometrial epithelial cells and embryonic trophoblast cells.

The extracellular vesicles (EVs) in uterine fluids play a vital role in embryo implantation by mediating intrauterine communication between conceptus and maternal endometrium in pigs. However, the regulatory mechanism of EVs in uterine fluids is largely unclear. In order to understand the effect of EVs in uterine flushing fluids (UFs) during embryo implantation on endometrial epithelial cells (EECs) and embryonic trophoblast cells (PTr2 cells). The UFs-EVs on day 13 of pregnancy (D13) were added to the culture medium of EECs and PTr2 cells. It was found that PKH-67 labeled UFs-EVs could be taken up in EECs and PTr2 cells. Transcriptome sequencing analysis showed that a total of 1793 and 6279 genes were differentially expressed in the EECs and PTr2 cells after the treatment of UFs-EVs on D13, respectively. Among these genes, real-time quantitative PCR (RT-qPCR) results indicated that ID2, ITGA5, CXCL10 and CXCL11 genes were differentially expressed in both EECs and PTr2 cells after treatment. Bioinformatics analysis showed that the differentially expressed (DE) genes in EECs and PTr2 cells after treatment are involved in immune regulation, cell migration, cell adhesion and the secretion and uptake of EVs. Our research offers novel insight into the regulation mechanism of UFs-EVs on D13 in EECs and PTr2 cells.

Ssc-miR-21-5p regulates endometrial epithelial cell proliferation, apoptosis and migration via the PDCD4/AKT pathway.

Endometrial receptivity plays a vital role in successful embryo implantation in pigs. MicroRNAs (miRNAs), known as regulators of gene expression, have been implicated in the regulation of embryo implantation. However, the role of miRNAs in endometrial receptivity during the pre-implantation period remains elusive. In this study, we report that the expression level of Sus scrofa (ssc)-miR-21-5p in porcine endometrium tissues was significantly increased from day 9 to day 12 of pregnancy. Knockdown of ssc-miR-21-5p inhibited proliferation and migration of endometrial epithelial cells (EECs), and induced their apoptosis. We verified that programmed cell death 4 (PDCD4) was a target gene of ssc-miR-21-5p. Inhibition of PDCD4 rescued the effect of ssc-miR-21-5p repression on EECs. Our results also revealed that knockdown of ssc-miR-21-5p impeded the phosphorylation of AKT (herein referring to AKT1) by targeting PDCD4, which further upregulated the expression of Bax, and downregulated the levels of Bcl2 and Mmp9. Furthermore, loss of function of Mus musculus (mmu)-miR-21-5p in vivo resulted in a decreased number of implanted mouse embryos. Taken together, knockdown of ssc-miR-21-5p hampers endometrial receptivity by modulating the PDCD4/AKT pathway.

Vitamin D status is associated with 1,5-anhydro-d-glucitol status in patients with type 2 diabetes mellitus.

Serum 1,5-anhydro-d-glucitol (1,5-AG) concentrations are short-term hyperglycemia indicators and were used to estimate the effects of serum vitamin D concentrations on glycemic control in patients with type 2 diabetes mellitus (T2DM). Serum concentrations of 1,5-AG, 25-hydroxyvitamin D2 (25-OH-D2), and 25-hydroxyvitamin D3 (25-OH-D3) from 11 026 patients with T2DM, hospitalized in the Department of Endocrinology, Shanghai Central Hospital of Xuhui District, from January 2012 to June 2015, were retrospectively analyzed. Correlation analyses revealed correlations between 1,5-AG and 25-OH-D3 (r = 0.05, P < 0.001), age (r = 0.05, P < 0.001), and 25-OH-D2 + 25-OH-D3 (25-OH-D2/D3) (r = 0.05, P < 0.001). Linear regression analyses revealed associations between 1,5-AG and 25-OH-D2/D3 (adjusted R2 = 0.003) as well as 25-OH-D3 (adjusted R2 = 0.002). In males with 1,5-AG levels ≤11.55 mg/L, serum concentrations of 25-OH-D2 (P < 0.001) and 25-OH-D3 (P = 0.001) were significantly lower than those in diabetic males with 1,5-AG levels >11.55 mg/L. Serum concentrations of 25-OH-D2/D3 in patients with T2DM were associated with 1,5-AG retention, suggesting involvement in glycemic control.

Overexpression of secretagogin promotes cell apoptosis and inhibits migration and invasion of human SW480 human colorectal cancer cells.

OBJECTIVE: In order to investigate the effect of secretagogin (SCGN) on colorectal cancer (CRC) cells apoptosis, invasion and migration in vitro. METHODS: Expression of SCGN in CRC tissues and the paired adjacent non-tumorous tissues (n = 36) and four human CRC cell lines (HT29, HCT116, SW480 and SW620) were detected. SW480 cells were transfected with the SCGN overexpression plasmid (eGFP-SCGN), si-SCGN-773, and the corresponding negative controls (NCs). Then, cell-cycle distribution, cell apoptosis, migration, invasion and expression of apoptosis- and metastasis-related proteins were detected. RESULTS: SCGN was significantly downregulated in CRC tissues as compared with the adjacent non-tumorous tissues. The expression of SCGN in HT29 and SW480 cells were lower than those in HT116 and SW620 cells. We transfected SW480 cells with SCGN overexpression plasmid eGFP-SCGN and found the increased cell apoptosis, with cell arresting at G0/G1 phase. SW480 cells with SCGN overexpression showed wider wound width and fewer invaded cells than control and blank cells, with upregulated Bax, cleaved Caspase 3 and E-cadherin, and downregulated Bcl-2 and Vimentin. We also transfected SW480 cells with si-SCGN-773 and found si-SCGN increased cell migration and invasion, but did not affect cell apoptosis and expression of related proteins. CONCLUSION: We concluded that the overexpression of SCGN in SW480 cells promoted cell apoptosis and inhibited cell migration and invasion.

Itraconazole attenuates hepatic gluconeogenesis and promotes glucose uptake by regulating AMPK pathway.

The primarily metabolic abnormality in type 2 diabetes mellitus (T2DM) is the defect in gluconeogenesis and glucose uptake. Itraconazole (ITCZ) is a traditional azole drug with anti-fungal and anticancer properties. However, limited attention has been directed towards the contribution of ITCZ to hepatic gluconeogenesis and glucose uptake in T2DM. The present study aimed to investigate the potential effects of ITCZ on hepatic gluconeogenesis and glucose uptake as well as the underlying mechanisms. No obvious change in cell viability was detected by MTT assay in HepG2 cells with ITCZ treatment at gradually increasing concentrations. Western blot analysis demonstrated that the phosphorylation level of 5' adenosine monophosphate-activated protein kinase (AMPK) was significantly elevated by ITCZ treatment at ≥5 µg/ml (P<0.05). Moreover, ITCZ repressed the gluconeogenesis of HepG2 cells, as evidenced by the dose-dependently increased glycogen synthase kinase 3ß phosphorylation level and a notably decreased glucose production rate (P<0.05). Simultaneously, the expression of peroxisome proliferator-activated receptor γ co-activator 1α, phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) in HepG2 cells was reduced by ITCZ in a dose-dependent manner (P<0.001). Furthermore, a 2-deoxyglucose uptake assay revealed that the glucose uptake of HepG2 cells was notably enhanced, accompanied by the ITCZ dose-dependent upregulation of glucose transporter-4 (GLUT-4) (P<0.05). Conversely, silencing of AMPK by small interfering RNA resulted in an increase of ITCZ-reduced gluconeogenesis and inhibition of ITCZ-induced glucose uptake with relative upregulation of PEPCK and G6Pase and downregulation of GLUT4 in the presence of 50 µg/ml ITCZ (P<0.05). Overall, the results indicated that AMPK has an important role in regulating ITCZ-induced glucose uptake by stimulating GLUT4 in HepG2 cells. Therefore, ITCZ may become a promising candidate for T2DM therapy.

Dipeptidyl peptidase 4 concentration influenced by serum insulin levels rather than arterial stiffness index in type 2 diabetics.

OBJECTIVE: To investigate whether inhibitors of dipeptidyl peptidase-4 (DPP-4) can suppress the atherosclerotic development in diabetic patients. METHODS: The prospective study was carried out in the Out-patient Department of XuHui Central Hospital, Shanghai, China, between March and August 2013. The correlation of major index of glucose and lipid metabolism profiles, and the arterial stiffness index (AI) between diabetic patients and healthy subjects were analyzed. PATIENTS OR MATERIALS: 39 patients with type 2 diabetes and 29 healthy subjects were enrolled for measurements of blood glucose, plasma insulin, HbA1c, TC, cholesterol, HDL, AI and body mass index (BMI). RESULTS: Significant differences were found between DPP-4 and blood glucose (fasting and 2 h postprandial), HbA1c, cholesterol, high density lipoprotein and arterial stiffness in normal subjects and diabetic patients. Only the fasting insulin concentration and high density lipoprotein had a significant impact on DPP-4 levels. CONCLUSIONS: It was clear that insulin (fasting) and HDL levels had an impact on DPP-4 activity but only in patients with Type 2 diabetes. The arterial stiffness index was not correlated with DPP-4 levels in Type 2 diabetic patients.

Association of the common rs9939609 variant of FTO gene with polycystic ovary syndrome in Chinese women.

Variations in the fat mass and obesity-associated (FTO) gene have recently been associated with obesity and type 2 diabetes mellitus among different ethnic populations. Given that the phenotype of polycystic ovary syndrome (PCOS) overlaps with obesity and type 2 diabetes, we hypothesize that the common rs9939609 variant of FTO gene is related to PCOS susceptibility. We performed a case-control association study on 215 women with PCOS, using 227 healthy women as the control. We examined the association between rs9939609 variant and PCOS susceptibility, as well as between PCOS and obesity-related parameters in Chinese women. We observed significant differences in the allelic and genotypic distributions between PCOS patients and the control group. The A allele was significantly more frequent among PCOS patients than in the control population (15.1% vs. 9.9%; A allele vs. T allele, OR = 1.62, P = 0.019). The A allele carrier genotype (AA and AT) frequencies were also significantly greater in PCOS patients than in the controls (28% vs. 19%; AT and TT vs. TT genotype, OR = 1·61, P = 0.035). In logistic regression, the strength of this association was attenuated after adjustment for body mass index (BMI) (A allele vs. T allele, OR = 1.39, P = 0.286; AT and TT genotypes vs. TT genotype, OR = 1.40, P = 0.312). However, we did not find any significant associations of rs9939609 variant with obesity-related traits. In conclusions, the rs9939609 variant in the FTO gene is associated with PCOS susceptibility in the Chinese population, probably because of its effect on BMI.

The association of TAAAAn repeat polymorphism in sex hormone-binding protein gene with polycystic ovary syndrome in Chinese population.

TAAAAn polymorphism in sex hormone-binding globulin (SHBG) gene has been found to influence the transcriptional activity of SHBG gene in vitro, and several studies have reported variable associations of this polymorphism with serum SHBG levels in women with polycystic ovary syndrome (PCOS). We have investigated the association of TAAAAn polymorphism with PCOS in 187 women with PCOS and 176 controls; in which five alleles (6-10 repeats) and 13 genotypes were found. None of the TAAAAn alleles or genotypes occurred in significant different frequency in PCOS patients compared with controls. Serum SHBG in the PCOS group was significantly lower than those in controls (33.8 +/- 30.2 nmol/l vs. 65.58 +/- 31.12 nmol/l; P < 0.01). Serum SHBG concentrations were similar for patients with PCOS whether they displayed short or long genotypes for TAAAAn (Log10SHBG: 1.46 +/- 0.38 and 1.58 +/- 0.33, respectively). Similar results were observed for controls (Log10SHBG: 1.79 +/- 0.17 and 1.77 +/- 0.14, respectively). In contrast, serum SHBG values were negatively associated with body mass index (BMI) and homeostasis model assessment of insulin resistance in the PCOS group (B = -0.285 and -0.264, respectively; P < 0.01). Thus, the TAAAAn polymorphism in SHBG gene was not a determinant of PCOS in this population of Chinese women, whereas, serum SHBG was significantly associated with BMI and insulin resistance in these PCOS patients.

Androgen receptor gene CAG(n) trinucleotide repeats polymorphism in Chinese women with polycystic ovary syndrome.

Trinucleotide repeats CAG(n) in androgen receptor gene is thought to be a potential site of genetic susceptibility to polycystic ovary syndrome (PCOS). However, previous studies of PCOS have shown variable association of CAG(n )polymorphism with PCOS. In order to evaluate CAG(n )polymorphism in Chinese women with PCOS, we have genotyped CAG(n) repeat numbers in female Chinese subjects (148 PCOS patients and 104 control subjects). The mean CAG(n) repeat lengths of PCOS patients and control subjects were similar (22.88 +/- 1.76 vs. 22.85 +/- 1.60; P = NS). No difference in the mean CAG(n) repeat lengths of hyperandrogenic and nonhyperandrogenic subgroups of PCOS patients was found (22.86 +/- 1.68 vs. 22.91 +/- 1.84; P = NS). Moreover, no difference was found in the term of mean CAG(n) repeat lengths in the nonhyperandrogenic subgroup and the control subjects (22.86 +/- 1.68 vs. 22.85 +/- 1.60; P = NS). However, mean CAG(n) repeat lengths were negatively correlated with serum total cholesterol and low-density lipoprotein-cholesterol concentration in PCOS patients (r = -0.182, P < 0.05 and r = -0.210, P < 0.05, respectively), but not with total testosterone, body mass index, waist and hip circumferences. The CAG(n) repeat length polymorphism may not be a major determinant of PCOS, but it may influence the lipid metabolism of PCOS patients.

Clinical and genetic analysis for a Chinese family with hereditary fructose intolerance.

Hereditary fructose intolerance (HFI) is an inheritable disorder of fructose metabolism, inherited as an autosomal recessive disorder and caused by catalytic deficiency of aldolase B, which is critical for gluconeogenesis and fructose metabolism. The affected individuals develop severe hypoglycemia after taking foods containing fructose and cognate sugars. The exons 2-9 of the aldolase B (gene symbol ALDOB) gene from one Chinese HFI patient were amplified by the polymerase chain reaction (PCR), and direct sequence determination was applied to the amplified fragments. The mutation of a 4-bp (AACA) deletion (479_482 del) in exon 4 of ALDOB gene was identified in the patient, which had been reported to cause a frameshift at codon 118 and a truncated protein of 132 amino acids in the previous study. Then, the second case with the same homozygote deletion and eight cases with heterozygotes had been found through screening for the mutation c.479_482 del AACA in the whole family. This is the first report of HFI with the mutation c.479_482 del AACA in the ALDOB gene in a Chinese family.