MiR-221-3p regulates the microvascular dysfunction in diabetic retinopathy by targeting TIMP3.

Diabetic retinopathy is one of the major complications of diabetes and the main cause to lead to blindness for diabetic patients. However, the exact mechanisms involved in the progression of diabetic retinopathy are not completely known. Herein, we demonstrated a novel role of miR-221-3p in the microvascular dysfunction in diabetic retinopathy. MiR-221-3p expression was found to be substantially upregulated in the retina samples of diabetic rats. Besides, ganglion cell layer, inner nuclear layer, outer nuclear layer, and retinal pigment epithelium layer of diabetic rats expressed higher miR-221-3p than the matched areas of normal rats. High glucose-treated retinal microvascular endothelial cells RF/6A and HRECs exhibited higher miR-221-3p than that in normal condition. MiR-221-3p inhibition could alleviate the retinal vascular leakage induced by diabetes in vivo as evaluated by Evans blue leakage assay, and reduce the proliferation, accelerate the apoptosis development, and inhibit the migration capacity of high glucose-treated RF/6A cells in vitro, while miR-221-3p overexpression partially enhanced the detrimental effects. By bioinformatics analysis and luciferase reporter assay, we identified that TIMP3 is the direct target of miR-221-3p. TIMP3 overexpression counteracted the effect of miR-221-3p on the vessel leakage and endothelial cell function. In conclusion, this study highlights the negative role of miR-221-3p in the microvascular dysfunction in diabetic retinopathy by targeting TIMP3, representing a potential therapeutic target for human diabetic retinopathy.

Genetic Architecture and Selection of Chinese Cattle Revealed by Whole Genome Resequencing.

The bovine genetic resources in China are diverse, but their value and potential are yet to be discovered. To determine the genetic diversity and population structure of Chinese cattle, we analyzed the whole genomes of 46 cattle from six phenotypically and geographically representative Chinese cattle breeds, together with 18 Red Angus cattle genomes, 11 Japanese black cattle genomes and taurine and indicine genomes available from previous studies. Our results showed that Chinese cattle originated from hybridization between Bos taurus and Bos indicus. Moreover, we found that the level of genetic variation in Chinese cattle depends upon the degree of indicine content. We also discovered many potential selective sweep regions associated with domestication related to breed-specific characteristics, with selective sweep regions including genes associated with coat color (ERCC2, MC1R, ZBTB17, and MAP2K1), dairy traits (NCAPG, MAPK7, FST, ITFG1, SETMAR, PAG1, CSN3, and RPL37A), and meat production/quality traits (such as BBS2, R3HDM1, IGFBP2, IGFBP5, MYH9, MYH4, and MC5R). These findings substantially expand the catalogue of genetic variants in cattle and reveal new insights into the evolutionary history and domestication traits of Chinese cattle.

C1qTNF-related protein 1 improve insulin resistance by reducing phosphorylation of serine 1101 in insulin receptor substrate 1.

C1qTNF-related protein 1 (CTRP1) is independently associated with type 2 diabetes. However, the relationship between CTRP1 and insulin resistance is still not established. This study aimed to explore the role of CTRP1 under the situation of insulin resistance in adipose tissue. Plasma CTRP1 level was investigated in type 2 diabetic subjects (n = 35) and non-diabetic subjects (n = 35). The relationship between CTRP1 and phosphorylation of multi insulin receptor substrate 1 (IRS-1) serine (Ser) sites was further explored. Our data showed that Plasma CTRP1 was higher and negative correlation with insulin resistance in diabetic subjects (r = -0.283, p = 0.018). Glucose utilisation test revealed that the glucose utilisation rate of mature adipocytes was improved by CTRP1 in the presence of insulin. CTRP1 was not only related to IRS-1 protein, but also negatively correlated with IRS-1 Ser1101 phosphorylation (r = -0.398, p = 0.031). Furthermore, Phosphorylation levels of IRS-1 Ser1101 were significantly lower after incubation with 40 ng/mL CTRP1 in mature adipocytes than those with no intervention (p < 0.05). There was no significant correlation between CTRP1 and other IRS-1 serine sites (Ser302, Ser307, Ser612, Ser636/639, and Ser789). Collectively, our results suggested that CTRP1 might improve insulin resistance by reducing the phosphorylation of IRS-1 Ser1101, induced in the situation of insulin resistance as a feedback adipokine.

Tissue expression analysis, cloning and characterization of the 5'-regulatory region of the bovine FABP3 gene.

Fatty acid binding protein 3 (FABP3) is a member of the FABP family which bind fatty acids and have an important role in fatty acid metabolism. A large number of studies have shown that the genetic polymorphisms of FABP3 are positively correlated with intramuscular fat (IMF) content in domestic animals, however, the function and transcriptional characteristics of FABP3 in cattle remain unclear. Real-time PCR analysis revealed that bovine FABP3 was highly expressed in cardiac tissue. The 5'-regulatory region of bovine FABP3 was cloned and its transcription initiation sites were identified. Sequence analysis showed that many transcriptional factor binding sites including TATA-box and CCAAT-box were present on the 5'-flanking region of bovine FABP3, and four CpG islands were found on nucleotides from -891 to +118. Seven serial deletion constructs of the 5'-regulatory region evaluated in dual-luciferase reporter assay indicated that its core promoter was 384 base pairs upstream from the transcription initiation site. The transcriptional factor binding sites RXRα, KLF15, CREB and Sp1 were conserved in the core promoter of cattle, sheep, pigs and dogs. These results provide further understanding of the function and regulation mechanism of bovine FABP3.

Elevated circulating levels of CTRP1, a novel adipokine, in diabetic patients.

Complement C1q tumor necrosis factor-related protein 1 (CTRP1), an adipose tissue-derived adipokine has been shown to decrease blood glucose levels and to improve metabolism of glucose in mice. In addition, CTRP1 has exhibited significant association with BMI, adiponectin and TNF-α in diabetic animal models. However, there are no published studies addressing CTRP1 levels in type 2 diabetic patients. Therefore, it was of interest to evaluate plasma CTRP1 levels and associated clinical parameters and biomarkers in patients with type 2 diabetes. 135 subjects were recruited to this study, including 62 type 2 diabetic patients (DM group) and 73 healthy subjects (control group). We measured biochemical parameters, CTRP1, TNF-α and adiponectin using enzyme-linked immunosorbent assay (ELISA). Plasma CTRP1 levels showed a significant difference between the DM group and the control group (646.3 ± 154.4 ng/mL vs. 442.6 ± 165.4 ng/mL, p < 0.01). In addition, CTRP1 was strongly positively associated with BMI, glucose levels, HbA1c, HOMA-IR and TNF-α in diabetic patients. CTRP1 showed negative correlation with adiponectin. In Multivariate regression analysis, CTRP1 was strongly independently associated with diabetes when CTRP1 levels were analyzed by both as a continuous variable and quartile (OR: 1.009, 95% CI: 1.004-1.015, p < 0.05; OR: 2.443, 95% CI: 1.379-4.182, p < 0.01, respectively). Increased plasma CTRP1 was independently associated with type 2 diabetes. Profiling of plasma adipokines such as CTRP1 is particularly important to obtain a greater understanding of their contribution to the type 2 diabetic state.

Introducing a new concept: Psychological capital of older people and its positive effect on mental health.

Objective: This study aimed to explore the structure of psychological capital (PsyCap) and its positive effects on mental health among older people. Methods: Study 1 used grounded theory to analyze the semi-structured interviewing data of 17 Chinese older people (60-96 years old) to develop a primary PsyCap questionnaire for older people. Study 2, respectively, applied exploratory factor analysis (EFA) with 198 Chinese older people (M = 69.2; SD = 6.685) and confirmatory factor analysis (CFA) with 370 Chinese older people (M = 73.84; SD = 9.416) to test a seven-factor structure for PsyCap. Study 3 used 328 participants (M = 79.73; SD = 9.073) to examine the correlation between PsyCap and mental health. Results: Study 1 identified that PsyCap of older people contains 'resilience,' 'self-efficacy,' 'optimism,' 'ease and content,' 'gratitude and dedication, 'wisdom,' and 'meaning in life' and generated a primary seven-factor questionnaire. Study 2 proved the overall and internal structure reliability of PsyCap were good (Cronbach's alphas ranged 0.809 ~ 0.935), and the seven-factor measurement model fitted the data well (χ 2/df = 2.07, RMSEA = 0.05, RMR = 0.05, CFI = 0.95, IFI = 0.95, TLI = 0.94, NFI = 0.91). The PsyCap scale was also proved to an excellent convergent validity, discriminant validity, calibration validity, and measurement invariance across different groups. Study 3 found that PsyCap and its seven factors significantly correlated with depression (r = -0.419 ~ -0.163, p < 0.01) after controlling the demographic variables. Conclusion: These findings provide a reliable and valid assessment for quantitative empirical research of PsyCap among older people and show significant impacts on mental health among older people, which offers new insight into improving mental health from the perspective of positive psychology.

Molecular characterization, expression pattern, polymorphism and association analysis of bovine ADAMTSL3 gene.

A disintegrin-like and metalloprotease domain with thrombospondin type I motifs-like 3 (ADAMTSL3) is an important candidate gene for body measurement traits through marker-assisted selection (MAS). The objective of this study was to analyze SNP of bovine ADAMTSL3 gene and its specific expression in tissues to explore its possible correlation with body measurement traits in Bos taurus. Our genomic structural analysis showed that bovine ADAMTSL3 shares much similarity with human ADAMTSL3. Gene expression analysis indicated that the order of specific tissue expression patterns of bovine ADAMTSL3 was the testis, heart, fat, stomach, small intestine, liver, lung, trachea, kidney, spleen, large intestine and muscle. Allele frequencies demonstrated significant variance in different local cattle breeds. Also, the T1532C SNP in exon 13 and C1899T SNP in exon 15 were significantly correlated with Body Length (BL), Chest Depth (CD), Heart Girth (HG) and Pin Bone Width (PBW). Furthermore, C1899T SNP marker had significant correlation with the PBW (P = 0. 0235) based on 867 individual analysis. In the total population, combination of T1532C and C1899T SNPs significantly affected on the BL (P = 0.0028), CD (P < 0.0001), HG (P = 0.0002) and PBW (P = 0.0049). These results suggest that the ADAMTSL3 gene, as one of target genes for body measurement traits in cattle breeds, could be used as a genetic marker to select excellent body type of cattle population in the animal breeding program.

Impacts of psychological resources, social network support and community support on social participation of older adults in China: Variations by different health-risk groups.

Social participation comprises comprehensive interactions between individual health and relevant environmental characteristics, which play an increasingly important role in promoting older adults' health and quality of life. Although many studies have examined impacts of various factors on the social participation of older adults, some internal mechanisms between health characteristics and environmental characteristics have not been revealed. This study aimed to examine the moderating effects of health characteristics on the relationships among psychological resources, social network support, community support and social participation of Chinese older adults. Based on some key health indicators, this study used the two-step cluster algorithm to divide the participants aged over 60 years from the Chinese Longitudinal Aging Social Survey in 2016 (N = 8669) into low (47.2%), moderate (33.8%) and high (18%) health risk groups. A multiple-group structural equation model was then built to examine the effects of psychological resources, social network support and community support on social participation across the three groups. The results suggested that with the increase in health risks, the contributions of psychological resources and community support to social participation decreased from 28.6% to 23.7% and from 61.5% to 50.7%, respectively, while that of social network support increased from 9.9% to 25.6%. These variations suggest that individual health has a significant moderating effect on the psychological and social compensating mechanisms of the social participation among Chinese older adults.

Insights into adaption and growth evolution: a comparative genomics study on two distinct cattle breeds from Northern and Southern China.

Mongolian cattle (MG, Bos taurus) and Minnan cattle (MN, Bos indicus) are two different breeds of Chinese indigenous cattle, representing North type and South type, respectively. However, their value and potential have not yet been discovered at the genomic level. In this study, 26 individuals of MN and MG were sequenced for the first time at an average of 13.9- and 12.8-fold, respectively. Large numbers of different variations were identified. In addition, the analyses of phylogenetic and population structure showed that these two cattle breeds are distinct from each other, and results of linkage disequilibrium analysis revealed that these two cattle breeds have undergone various degrees of intense natural or artificial selection. Subsequently, 496 and 306 potential selected genes (PSRs) were obtained in MN and MG, containing 1,096 and 529 potential selected genes (PSGs), respectively. These PSGs, together with the analyzed copy number variation (CNV)-related genes, showed potential relations with their phenotypic characteristics, including environmental adaptability (e.g., DVL2, HSPA4, CDHR4), feed efficiency (e.g., R3HDM1, PLAG1, XKR4), and meat/milk production (e.g., PDHB, LEMD3, APOF). The results of this study help to gain new insights into the genetic characteristics of two distinct cattle breeds and will contribute to future cattle breeding.

Population genetic analysis of 6 Y-STR loci in Chinese northwestern Qinchuan yellow cattle breed.

Six Y-STR loci (UMN0929, UMN0108, UMN0920, INRA124, UMN2404 and UMN0103) were analyzed using 576 healthy and unrelated males and 10 females of the Qinchuan cattle population in Chinese Shaanxi Province. Allele frequency, gene diversity, the polymorphic information content, and the number of effective gene were calculated. All loci were in accordance with the Hardy-Weinberg equilibrium (P > 0.05). The population data were compared with published data of other cattle breeds, suggesting that Qinchuan cattle were originated primarily from Bos Taurus. Results are valuable for individual identification, paternity testing, and origin analysis of Qinchuan cattle breed.

Molecular characterization, polymorphism of bovine ZBTB38 gene and association with body measurement traits in native Chinese cattle breeds.

Zinc finger and BTB domain containing 38 (ZBTB38), binding to and repressing methylated DNA, is an important candidate gene for selection of body measurement traits through marker-assisted selection (MAS). The expression of ZBTB38 is regulated in human and animal height as well as other stature indexes. Genomic structural analysis shows that bovine ZBTB38 shares much similarity with human ZBTB38. We discovered and evaluated the potential association of the single nucleotide polymorphism (SNP) of the bovine ZBTB38 gene with body measurement traits in 722 individuals. The latest findings demonstrate that the A841G SNP in exon 1 is significantly associated with Body Length (BL), Hip Height (HH) and Heart Girth (HG). Furthermore, the analysis of A841G SNP marker shows that there are significant effects on the BL (P = 0.0389) in 722 individuals, significant effects on the HH (P = 0.0173) and HG (P = 0.0147) in Qinchuan improvement steers (QI) population, as well as significant effects on the WH (P = 0.0094) in Xuelong (XL) population. These results clearly suggest that the ZBTB38 gene is among of target genes for body measurement traits in bovine reproduction and breeding, and thus provide data for establishment of an animal model using cattle to study big animal body type.

Increased TRPC6 expression is associated with tubular epithelial cell proliferation and inflammation in diabetic nephropathy.

Although TRPC6 expression is shown to be significantly elevated in a rat model diabetic nephropathy (DN), its expression and role in human DN are unclear. We thus explored the role of TRPC6 in the pathophysiology of tubular epithelial cell injury following DN. HK-2 cells were cultured in a high-glucose medium to induce a DN cell model. Ad-TRPC6 and TRP6 siRNA were transfected to overexpress and knock down TRPC6. We found that TRPC6 expression was significantly upregulated in DN tissues and cells. TRPC6 siRNA inhibited cell proliferation and promoted cell apoptosis in HK-2 cells treated with high glucose, whereas Ad-TRPC6 showed the opposite effect. Furthermore, Ad-TRPC6 significantly promoted release of IL-8 and IL-6. Subsequent experiments demonstrated that the signaling pathway of nuclear factor of activated T cells (NFAT) was activated by Ad-TRPC6 and deactivated by TRPC6 siRNA. The NFAT signaling inhibitor, FK-506, eliminated the effect of TRPC6 on HK-2 cells. These results suggest that TRPC6 was upregulated in DN and could promote cell proliferation and inflammation by inhibiting the NFAT signaling pathway in tubular epithelial cells.

Molecular cloning of bovine FABGL gene and its effects on bovine bioeconomic traits.

The complete CDS sequence of the bovine FABGL gene was determined by homology cloning approach combined with RT-PCR and 3'- and 5'-RACE. The results of sequence analysis and bioinformatics study showed that this cDNA contained 994 nucleotides, with a 780 bp open reading frame (ORF) flanked by a 16 bp 5'-UTR (incompletely) and a 198 bp 3'-UTR. The deduced amino acid sequence (260 AA) shows 88% identity with the corresponding sequence in humans. Two single nucleotide substitutions, one located in intron 5 (I5) at position 1 065 bp (Y = C/T) (GenBank: DQ409814) and the other in intron 8 (I8) at position 1 792 bp (R = A/G), were detected using the PCR-SSCP method. Analysis of the allele frequencies of the two polymorphic sites in three different cattle breeds (Angus, Hereford, and Simmental) with different genotypes showed large differences: in locus I8, cattle with the GG genotype showed higher beef performance index (BPI) (4.283 +/- 0.475 kg/cm) in comparison with cattle with the AA genotype (4.008 +/- 0.465 kg/cm) (P = 0.01). Regarding the ribeye area, cattle with the GG genotype showed significantly higher ribeye area (73.380 +/- 13.005 cm(2)) compared with cattle with the AA genotype (67.744 +/- 12.777 cm(2)) (p = 0.05). In locus I5, some associations for the average daily gain (ADG) were found at the significance level of 0.01 between three different genotypes (CC, CT, TT): cattle with the TT genotype showed the highest ADG (0.652 +/- 0.330 kg/d), whereas cattle with the CC genotype showed the lowest ADG value (0.421 +/- 0.178 kg/d).

Whole-genome sequencing of the endangered bovine species Gayal (Bos frontalis) provides new insights into its genetic features.

Gayal (Bos frontalis) is a semi-wild and endangered bovine species that differs from domestic cattle (Bos taurus and Bos indicus), and its genetic background remains unclear. Here, we performed whole-genome sequencing of one Gayal for the first time, with one Red Angus cattle and one Japanese Black cattle as controls. In total, 97.8 Gb of sequencing reads were generated with an average 11.78-fold depth and >98.44% coverage of the reference sequence (UMD3.1). Numerous different variations were identified, 62.24% of the total single nucleotide polymorphisms (SNPs) detected in Gayal were novel, and 16,901 breed-specific nonsynonymous SNPs (BS-nsSNPs) that might be associated with traits of interest in Gayal were further investigated. Moreover, the demographic history of bovine species was first analyzed, and two population expansions and two population bottlenecks were identified. The obvious differences among their population sizes supported that Gayal was not B. taurus. The phylogenic analysis suggested that Gayal was a hybrid descendant from crossing of male wild gaur and female domestic cattle. These discoveries will provide valuable genomic information regarding potential genomic markers that could predict traits of interest for breeding programs of these cattle breeds and may assist relevant departments with future conservation and utilization of Gayal.

Overexpression of Wilms tumor 1 gene as a negative prognostic indicator in acute myeloid leukemia.

Chromosomal aberrations are useful in assessing treatment options and clinical outcomes of acute myeloid leukemia (AML) patients. However, 40 ∼ 50% of the AML patients showed no chromosomal abnormalities, i.e., with normal cytogenetics aka the CN-AML patients. Testing of molecular aberrations such as FLT3 or NPM1 can help to define clinical outcomes in the CN-AML patients but with various successes. Goal of this study was to test the possibility of Wilms' tumor 1 (WT1) gene overexpression as an additional molecular biomarker. A total of 103 CN-AML patients, among which 28% had overexpressed WT1, were studied over a period of 38 months. Patient's response to induction chemotherapy as measured by the complete remission (CR) rate, disease-free survival (DFS) and overall survival (OS) were measured. Our data suggested that WT1 overexpression correlated negatively with the CR rate, DFS and OS. Consistent with previous reports, CN-AML patients can be divided into three different risk subgroups based on the status of known molecular abnormalities, i.e., the favorable (NPM1(mt)/no FLT3(ITD)), the unfavorable (FLT3(ITD)) and the intermediate risk subgroups. The WT1 overexpression significantly reduced the CR, DFS and OS in both the favorable and unfavorable groups. As the results, patients with normal WT1 gene expression in the favorable risk group showed the best clinical outcomes and all survived with complete remission and disease-free survival over the 37 month study period; in contrast, patients with WT1 overexpression in the unfavorable risk group displayed the worst clinical outcomes. WT1 overexpression by itself is an independent and negative indicator for predicting CR rate, DFS and OS of the CN-AML patients; moreover, it increases the statistical power of predicting the same clinical outcomes when it is combined with the NPM1(mt) or the FLT3(ITD) genotypes that are the good or poor prognostic markers of CN-AML.

Proopiomelanocortin gene polymorphisms and its association with meat quality traits by ultrasound measurement in Chinese cattle.

Ultrasound technology was used to measure live animal meat traits instead of true carcass meat traits for beef production and cattle breeding by an increasing number of institutions. In this study, we analyzed the association between genetic polymorphisms of proopiomelanocortin (POMC) and ultrasound measurement traits in Chinese cattle. Using direct DNA sequencing in 322 individuals of 7 different cattle subpopulation, 7 SNPs were identified for genotyping within 790bp region of intron 2 and exon 3 of POMC. 6586 T>G in intron 2 and 6769 C>T and 7216 C>T in exon 3 were significantly associated with ultrasound backfat thickness (UBF) (P<0.05) and ultrasound loin muscle area (ULMA) (P<0.01) in the total population; 6694 C>T, 6706 T>C, 6796 C>T and 6810 C>T in exon 3 were significantly associated with ULMA (P<0.0001) in the total population. These results clearly suggest that these SNPs of POMC be benefit for selection of individuals with good quality meat in Chinese cattle breeding program. Following validation in other populations and breeds, these markers could be incorporated into breeding programs to increase the rate of improvement in carcass and meat quality traits.

ZBTB38 gene polymorphism associated with body measurement traits in native Chinese cattle breeds.

Body measurement traits, influenced by genes and environmental factors, play numerous important roles in the value assessment of productivity and economy. In this study, we investigated the association between genetic polymorphisms of the zinc finger and BTB domain containing 38 gene (ZBTB38) and body measurement traits in native Chinese cattle. Using direct DNA sequencing in 423 individuals of 8 different cattle subpopulations, 9 novel polymorphisms were identified for genotyping within 647 bp region of exon 1 of ZBTB38. Linkage disequilibrium and association analysis revealed that two coding exon polymorphisms (2323 G>A and 2325 C>T polymorphisms), missense mutations valine GTC(T) to isoleucine ATC(T), were associated with body length (BL), withers height (WH) and rump length (RL). Furthermore, the analysis of 2323 G>A and 2325 C>T SNP markers shows that there are significant effects on the BL (P=0.0488), WH (P=0.0044) and RL (P=0.0314) in the total population. These results clearly suggest that the ZBTB38 gene is among the target genes for body measurement traits in bovine breeding, and provide data for establishment of an animal model using cattle to study big animal body type.

The cryoprotective effects of ascorbic acid supplementation on bovine semen quality.

To investigate the effects of ascorbic acid supplementation on standard semen quality parameters and antioxidant activities after thawing of bovine frozen semen, antioxidant ascorbic acid was added at concentrations of 2.5, 4.5, 6.5 and 8.5 mg/ml to bovine semen cryoprotective medium. The results showed that the sperm motility and motion characteristics were improved in the presence of ascorbic acid in extender, as compared to the control. The motility and straight linear velocity (VSL), linearity index (LIN), average path velocity (VAP), wobble coefficient (WOB), lateral head displacement (ALH) values and the percentage of "grade A" sperm in the extender supplemented with 4.5 mg/ml ascorbic acid were significantly higher than that of other treatment groups (P<0.05). The acrosome integrity and membrane integrity were significantly improved (P<0.05) by supplementing with 4.5 mg/ml ascorbic acid in the extender compared with a control. The extender supplemented with ascorbic acid did not lead to any improvement in superoxide dismutase (SOD) levels. The catalase (CAT) activity was higher in the extender supplemented with ascorbic acid at 4.5 mg/ml, when compared with other groups (P<0.05) and the extender supplemented with ascorbic acid significantly decreased glutathione peroxidase (GSH-Px) activity, whereas reduced glutathione (GSH) activities were significantly enhanced, compared with the control (P<0.05). Increasing the doses level of ascorbic acid decreased GSH-Px and GSH activity, the supplementation of 8.5 mg/ml ascorbic acid produced the lowest level of GSH-Px and GSH activity among groups (P<0.05). The extender supplemented with ascorbic acid could reduce the oxidative stress provoked by freezing-thawing and improve bovine semen quality. The particular properties of ascorbic acid are poorly related to its effectiveness in membrane cryopreservation. Further studies are required to determine lipid peroxidation and antioxidant capacities of ascorbic acid in cryopreserved bovine semen.