Regional difference assessment of the South-to-North Water Diversion project on improving intensive utilization efficiency of water resources in the water-receiving areas.

Improving the intensive utilization efficiency of water resources is essential to promote the sustainable utilization and management of water resources in water shortage areas, such as the water-receiving areas of water transfer project. Since the middle line project of South-to-North Diversion (SNWD) officially put into operation in 2014, the status of water resource supply and management in the water-receiving areas in China has changed. This study aimed to analyze the impact of the middle line project of SNWD on the intensive utilization efficiency of water resources, combined with the analysis results in the effects of the middle line project of SNWD under a variety of heterogeneous factors, so as to provide a policy reference for the utilization and management of water resources in water-receiving areas. Specifically, the BCC model based on the input perspective was adopted to calculate the water resource intensive utilization efficiency in 17 cities of Henan Province in China from 2011 to 2020. On this basis, the regional differences in the effects of the middle line project of SNWD on the water resource intensive utilization efficiency were analyzed through the difference-in-differences (DID) method. The results indicated that (1) during the study period, the average value of water resource intensive utilization efficiency in water-receiving areas was higher than that in non-water-receiving areas in Henan province, and the development trends of them showed "U"-shaped characteristics. (2) The middle line project of SNWD significantly has promoted the water resource intensive utilization efficiency in the water-receiving areas in Henan Province. (3) The heterogeneous differences of economic development, opening-up level, government influence, water resource endowment, and water resource policies would lead to regional differences in the effect of the middle line project of SNWD. Therefore, the government should adopt differentiated policies to improve the intensive utilization efficiency of water resources in accordance with the development conditions of the water-receiving areas.

Decoding Single-cell Landscape and Intercellular Crosstalk in the Transplanted Liver.

BACKGROUND: Liver transplantation (LT) is the most effective treatment for various end-stage liver diseases. However, the cellular complexity and intercellular crosstalk of the transplanted liver have constrained analyses of graft reconstruction after LT. METHODS: We established an immune-tolerated orthotopic LT mouse model to understand the physiological process of graft recovery and intercellular crosstalk. We employed single-cell RNA sequencing and cytometry by time-of-flight to comprehensively reveal the cellular landscape. RESULTS: We identified an acute and stable phase during perioperative graft recovery. Using single-cell technology, we made detailed annotations of the cellular landscape of the transplanted liver and determined dynamic modifications of these cells during LT. We found that 96% of graft-derived immune cells were replaced by recipient-derived cells from the preoperative to the stable phase. However, CD206 + MerTK + macrophages and CD49a + CD49b - natural killer cells were composed of both graft and recipient sources even in the stable phase. Intriguingly, the transcriptional profiles of these populations exhibited tissue-resident characteristics, suggesting that recipient-derived macrophages and natural killer cells have the potential to differentiate into 'tissue-resident cells' after LT. Furthermore, we described the transcriptional characteristics of these populations and implicated their role in regulating the metabolic and immune remodeling of the transplanted liver. CONCLUSIONS: In summary, this study delineated a cell atlas (type-proportion-source-time) of the transplanted liver and shed light on the physiological process of graft reconstruction and graft-recipient crosstalk.

Role of variation in platelet indices in pregnancy-related pathological complications and the prediction of postpartum hemorrhage: a case-control study.

BACKGROUND: Pregnancy-related pathological complications (PPCs) increase the risk of postpartum hemorrhage (PPH), Platelet activation and destruction are expected outcomes of PPCs. This study sought to compare the platelet indices of non-pregnant (NP) child-bearing aged women, healthy pregnant (HP) women, and women with PPCs, and investigate the use of these indices in PPH prediction. METHODS: This retrospective clinical study included 260 NP child-bearing aged women and 119 pregnant women. Of the 119 pregnant patients, 69 had HPs and 50 suffered from PPCs. Further, 50 patients delivered with PPH. We compared the platelet counts (PCs), mean platelet volumes (MPVs), platelet distribution widths (PDWs), plateletcrits (Pcts), MPV ratios (PC/MPV and Pct/MPV), alpha angles (angles), and maximum amplitudes (MAs) of the patients using Sysmex XN10 hematology analyzer and TEG 5000 Hemostasis analyzer system, respectively. RESULTS: With the exception of PDW, there were significant differences in the platelet parameters of the NP, HP, and PPC patients (P<0.05). The intergroup comparison results showed that the NP patients differed significantly from the HP and PPC patients in terms of age, MA, PC, Pct, and Pct/MPV (P<0.0125). Further, the HP and PPC patients differed significantly in terms of Pct, MPV, PC/MPV, and Pct/MPV (P<0.0125). Additionally, the univariate analysis showed that in the PPC patients, low MPV values were strongly related to PPH [odds ratio (OR) =0.012, P=0.003; OR =0.331, P=0.047]. CONCLUSIONS: Women with PPCs had significantly lower PC, Pct, PC/MPV and Pct/MPV values, but significantly higher MA and MPV values. PPHs were strongly related to PPC and low MPV values. A timely accurate diagnosis and evaluating MPV values may be useful in the prediction of PPH.

Prevention of platelet transfusion refractoriness by platelet antigen gene matching: a systematic review and meta-analysis.

BACKGROUND: To investigate the prevention of platelet transfusion refractoriness (PTR) by platelet antigen gene matching using literature search and meta-analysis. METHODS: PubMed (2000.1-2021.8), Embase (2000.1-2021.8), Cochrane (2010.1-2021.8), and the Chinese Biomedical Literature Database CBM (2010.1-2021.8) were selected as the search database platform. The keywords (HLA/Human leukocyte antigen), (HPA/Human platelet alloantigens), (genotyping/cross-match), platelet transfusion (PLT), and (CCI/Corrected Count Increment) were used for the joint search. After the literature was screened for inclusion and exclusion criteria, the Cochrane intervention handbook was used for bias risk assessment, and Revman 5.3.5 software was used for analysis to obtain the statistical forest plot and funnel plot. RESULTS: The preliminary results revealed 255 publications, and seven (297 patients in total) were finally included in the quantitative analysis. A total of five publications reported comparison of the 1 h CCI index of HLA or HPA gene matching and PLT after random selection, and the heterogeneity test showed statistical difference (I2=49%, P=0.10). The combined statistical analysis results were: (MD =8.57, 95% CI: 7.30-9.80, Z=13.30, P<0.00001), and while six publications reported the effective rate index of PLT, and the heterogeneity test showed no statistical difference (I2=43%, P=0.12). The fixed effect mode was used to compare the effective rate of the two intervention methods (OR =4.90, 95% CI: 3.50-6.86, Z=9.23, P<0.00001). DISCUSSION: HLA or HPA gene matching can improve the increment after PLT and reduce the incidence of ineffective PLT.

Alterations of the Gut Microbiome and Metabolome in Patients With Proliferative Diabetic Retinopathy.

Diabetic retinopathy (DR) has been reported to associate with gut microbiota alterations in murine models and thus "gut-retina-axis" has been proposed. However, the role of gut microbiome and the associated metabolism in DR patients still need to be elucidated. In this study, we collected fecal samples from 45 patients with proliferative diabetic retinopathy (PDR) and 90 matched diabetic patients (1:2 according to age, sex, and duration of diabetes) without DR (NDR) and performed 16S rRNA gene sequencing and untargeted metabolomics. We observed significantly lower bacterial diversity in the PDR group than that in the NDR group. Differential gut bacterial composition was also found, with significant depletion of 22 families (e.g., Coriobacteriaceae, Veillonellaceae, and Streptococcaceae) and enrichment of two families (Burkholderiaceae and Burkholderiales_unclassified) in the PDR group as compared with the NDR group. There were significantly different fecal metabolic features, which were enriched in metabolic pathways such as arachidonic acid and microbial metabolism, between the two groups. Among 36 coabundance metabolite clusters, 11 were positively/negatively contributed to PDR using logistic regression analysis. Fifteen gut microbial families were significantly correlated with the 11 metabolite clusters. Furthermore, a fecal metabolite-based classifier was constructed to distinguish PDR patients from NDR patients accurately. In conclusion, PDR is associated with reduced diversity and altered composition of gut microbiota and specific microbe-metabolite interplay. Our findings help to better understand the disease pathogenesis and provide novel diagnostic and therapeutic targets for PDR.

Sarcomatoid hepatocellular carcinoma: From clinical features to cancer genome.

BACKGROUND: Sarcomatoid hepatocellular carcinoma (HCC) is a rare and highly lethal histological subtype of HCC, with completely unknown genetic etiology and therapeutic targets. METHODS: We included 16 patients with sarcomatoid HCC receiving radical resection among 6731 cases of pathological confirmed HCC in year 2008 to 2018 in our hospital. We compared the clinical features, prognosis and cancer genome between 15 sarcomatoid HCC and propensity score-matched 75 non-sarcomatoid HCC patients. The other concurrent case was analyzed using phylogenetic tree to assess the tumor heterogeneity and evolution. RESULTS: Sarcomatoid HCC group showed larger tumor size, more advanced differentiation grade, lower tumor free survival (p = 0.038) and overall survival (p = 0.001), and sarcomatoid type was an independent risk factor for patient death. Integrating sarcomatoid subtype into AJCC staging could increase the diagnostic curve in predicting patient survival. The cancer genome spectrum showed sarcomatoid HCC group had significant higher mutation rates in CDKN2A, EPHA5, FANCM and MAP3K1. Mutations in CDKN2A significantly reduced tumor-free and overall survival in sarcomatoid HCC patients. Moreover, 46.6% sarcomatoid HCC patients had druggable mutations in cell cycle pathway genes, which were targeted by Abemaciclib, et al. We also found sarcomatoid and non-sarcomatoid lesions might originate from a common progenitor but progress differently. CONCLUSION: Our cancer genome analysis showed a specific genomic profile of sarcomatoid HCC, which were characterized by a high mutation rate in cell cycle genes particularly CDKN2A. The results indicate CDK4/6 inhibitors including abemaciclib, ribociclib and palbociclib as potential therapeutic targets and may help for therapeutic decision making.

Comparative genomic analysis of head and body/tail of pancreatic ductal adenocarcinoma at early and late stages.

Pancreatic ductal adenocarcinoma (PDAC), one of the most lethal human cancers, can be divided into head and body/tail cancers anatomically. We previously reported a prognostic relevance of tumour location in resectable PDAC. This study aimed to further explore the mechanism underlying the molecular diversity between the head and body/tail of PDACs. We detected tumour genomes in 154 resectable (surgery) and non-resectable (biopsy) PDACs using a next-generation sequencing panel. Wilcoxon's rank test or Fisher's exact test was used for evaluating associations between clinical characteristics, mutation frequency and survival probability between the two cohorts. Compared with pancreatic head cancers, pancreatic body/tail cancers showed significantly more enriched genomic alterations in KRAS (97.1% vs 82.4%, P = 0.004) and SMAD4 (42.0% vs 21.2%, P = 0.008). At early stages (I-II), the SMAD4 mutation rate was significantly higher in pancreatic body/tail cancers than pancreatic head cancers (56.0% vs 26.5%, P = 0.021). At late stages (III-IV), pancreatic body/tail cancers presented significantly higher KRAS mutation rate (100.0% vs 75.8%, P = 0.001), higher frequency of MAPK pathway mutation (100% vs 87.8%, P = 0.040) and lower rates of druggable genomic alterations (30.8% vs 57.6%, P = 0.030) than pancreatic head cancers. Our work points out that pancreatic body/tail cancer seems to be more malignant than pancreatic head cancer at late stages.

A model integrating donor gene polymorphisms predicts fibrosis after liver transplantation.

Post-transplant liver fibrosis (PTLF) is a common and severe complication in liver recipients. In this study, we assessed the impact of donor liver genetics on the development of PTLF. A total of 232 patients undergoing liver transplantation were included. Twenty-two single nucleotide polymorphisms (SNPs) associated with liver fibrosis were analyzed. Univariate analysis revealed seven donor SNPs to be associated with PTLF. In a multivariate analysis, independent risk factors of PTLF were genetic variation of donor GRP78 rs430397 (OR = 8.99, p = 0.003), GSTP1 rs1695 (OR = 0.13, p = 0.021), miRNA-196a rs12304647 (OR = 16.01, p =0.001), and TNF-α rs1800630 (OR = 79.78, p = 0.001); blood tacrolimus levels at maintenance > 7 ng/ml (OR =7.48, p <0.001); and post-transplant diabetes mellitus (OR = 7.50, p = 0.001). A predictive model that included donor SNPs showed better prognostic ability for PTLF than a model with only clinical parameters (AUROC: 0.863 vs 0.707, P < 0.001). Given that donor gene SNPs are associated with an increased risk of PTLF, this model integrated with donor gene polymorphisms may help clinicians predict PTLF.

The Similar Effects of miR-512-3p and miR-519a-2-5p on the Promotion of Hepatocellular Carcinoma: Different Tunes Sung With Equal Skill.

Although the therapeutic methods of hepatocellular carcinoma (HCC) have made great advances, the current situation is that HCC is the common malignancy. Our previous bioinformatic study presented that two members of C19MC (mir-512-1 and mir-519a-2) acted as crucial roles in the HCC progression. In this study, we first demonstrated that the miR-512-3p and miR-519a-2-5p, which were spliced from the mir-512-1 and mir-519a-2, were the functional mature miRNAs. Meanwhile, both miR-512-3p and miR-519a-2-5p were significantly upregulated in human HCC samples and HCC cell lines. The miR-512-3p and miR-519a-2-5p promoted the proliferation, invasion, and metastasis in vitro and in vivo. Moreover, the two miRNAs co-targeted the downstream tumor suppressors MAP3K2 and MAP2K4 and subsequently achieved the HCC progression. In the clinical cohort, high expression of miR-512-3p and miR-519a-2-5p acted as two risk factors for HCC recurrence and distinguished patients with poor tumor-free survival after radical resection. The integration of the two miRNAs into the AJCC staging system significantly improved the accuracy for the prediction of HCC recurrence. Our study suggests that miR-512-3p and miR-519a-2-5p have similar effects on the promotion of HCC progression. They can be robust markers for the prediction of HCC recurrence and therapy targets.

TCF7L2 rs290487 C allele aberrantly enhances hepatic gluconeogenesis through allele-specific changes in transcription and chromatin binding.

In this study, we investigated the mechanisms underlying the altered hepatic glucose metabolism and enhanced diabetes risk in individuals with the TCF7L2 rs290487 C allele. Analysis of 195 cirrhotic patients revealed a higher insulin resistance index and incidence of hepatogenous diabetes in patients with the rs290487 C/C genotype compared to those with the C/T or T/T genotype. The in vitro experiments using targeted mutant PLC-PRF-5 cell line showed that cells with the rs290487 C/C genotype (C/C cells) had higher glucose production, lower glucose uptake, and lower TCF7L2 mRNA and protein levels than those with the C/T genotype (C/T cells). Integrated multi-omics analysis of ChIP-seq, ATAC-seq, RNA-seq, and metabolomics data showed genome-wide alterations in the DNA binding affinity of TCF7L2 in the C/C cells, including gain (e.g., PFKP and PPARGC1A) and loss (e.g., PGK1 and PGM1) of binding sites in several glucose metabolism-related genes. These allele-specific changes in transcriptional regulation lead to increased expression of gluconeogenesis-related genes (PCK1, G6PC and PPARGC1A) and their downstream metabolites (oxaloacetate and ß-D-fructose 2,6-bisphosphate). These findings demonstrate that the TCF7L2 rs290487 C allele enhances gluconeogenesis through allele-specific changes in transcription and chromatin binding.

Predicting dyslipidemia after liver transplantation: A significant role of recipient metabolic inflammation profile.

BACKGROUND: Post-transplant dyslipidemia (PTDL) is a common complication in liver recipients and can cause morbidity and threaten graft function. The crosstalk between metabolic inflammation and dyslipidemia has been recently revealed. However, the role of grafts' and recipients' metabolic status in the development of PTDL has not been evaluated. AIM: To investigate the association of recipients' metabolic inflammation status with PTDL and construct a predictive model. METHODS: A total of 396 adult patients who received primary liver transplantation between 2015 and 2017 were enrolled. Metabolomics and cytokines were analyzed using recipients' pre-transplant peripheral blood in a training set (n = 72). An integrated prediction model was established according to the clinical risk factors and metabolic inflammation compounds and further verified in a validation set (n = 144). RESULTS: The serum lipid profile took 3 mo to reach homeostasis after liver transplantation. A total of 278 (70.2%) liver recipients developed PTDL during a follow-up period of 1.78 (1.00, 2.97) years. The PTDL group showed a significantly lower tumor-free survival and overall survival than the non-PTDL group in patients with hepatocellular carcinoma (n = 169). The metabolomic analysis showed that metabolic features discriminating between the PTDL and non-PTDL groups were associated with lipid and glucose metabolism-associated pathways. Among metabolites and cytokines differentially expressed between the two groups, interleukin-12 (p70) showed the best diagnostic accuracy and significantly increased the predictive value when it was incorporated into the clinical model in both training and validation sets. CONCLUSION: Recipients' pre-transplant serum interleukin-12 (p70) level is associated with the risk of PTDL and has potential clinical value for predicting PTDL.

The mir-767-105 cluster: a crucial factor related to the poor prognosis of hepatocellular carcinoma.

MiRNAs have been widely reported as the therapeutic target for hepatocellular carcinoma (HCC). However, mirna clusters, as the more impressive tumor regulatory factors, have received little attention. By deeply digging the Cancer Genome Atlas (TCGA) database, we aimed to explore the vital mirna cluster that regulated the poor prognosis of HCC. The results showed that the upregulation of mirna cluster-767-105 in HCC was the most significant, compared with the non-tumor tissues. Besides, high expression of all three members of the cluster was positively correlated with poor prognosis of HCC and the resistance of sorafenib. Cox analysis proved that all the three mirnas were independent prognostic factors, while the mir-767 was the most compelling (HR value 8.388, 95%CI 2.524-27.897). The higher expression of the three-mirna signature also significantly indicated the worse prognosis. Through bioinformatics analysis, we screened their common potential target genes, which were highly correlated with tumor regulation. These results supported that the mirna cluster-767-105 promoted the poor outcome of HCC and could be a robust target for the therapy of HCC patients.

The chromosome 19 microRNA cluster, regulated by promoter hypomethylation, is associated with tumour burden and poor prognosis in patients with hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is still one of the major malignant tumours with poor prognosis. The chromosome 19 microRNA cluster (C19MC) is the largest miRNA cluster, and its functions and regulatory mechanisms remain unclear in HCC. We extracted data from 373 HCC samples and 50 non-tumour samples from The Cancer Genome Atlas database. The differential expression levels and methylation levels of C19MC as well as the correlation between them were analysed. We evaluated the correlation between the expression levels of C19MC and the clinical features. We further performed prognostic analysis for C19MC and analysed the bioinformatic function. C19MC had upregulated expression levels and promoter hypomethylation in HCC. A significant negative correlation between the high expression and low methylation level of C19MC was obtained. In addition, the positive correlation between the expression levels of C19MC and the tumour grade, tumour stage and T-stage is shown. Three miRNAs (mir-512-1, mir-516a-1, mir-519a-2) were negatively associated with overall survival on the basis of the Kaplan-Meier analysis and the 3-miRNA signature was significant for the prognostic assessment of HCC. A bioinformatic enrichment analysis suggested that the target genes of the 3 miRNAs may be associated with mitogen-activated protein kinase pathways related to cancer invasion. In summary, our novel study demonstrated that the hypomethylation of promoters upregulates the expression levels of C19MC and that C19MC may represent a potential new candidate for the diagnosis and therapy of HCC.

A Path-Integral-Based Reinforcement Learning Algorithm for Path Following of an Autoassembly Mobile Robot.

Reinforcement learning (RL) combined with deep neural networks has led to a number of great achievements for robot control in virtual computer environments, where sufficient data can be obtained without any difficulty to train various models. However, thus far, only few and relatively simple tasks have been accomplished for practical robots, which is mainly caused by the following two reasons. First, training with real robots, especially with dynamic systems, is too complicated to be fully and accurately represented in simulations. Second, it is very costly to obtain training data from real systems. To address these two problems effectively, in this article, a path-integral-based RL algorithm is proposed for the task of path following of an autoassembly mobile robot, wherein three kernel techniques are introduced. First, a generalized path-integral-control approach is proposed to obtain the numerical solution of a stochastic dynamical system, wherein the calculation of the gradient and kinematics inverse is avoided to ensure fast and reliable training convergence. Second, a novel parameterization method using Lyapunov techniques is introduced into the RL algorithm to ensure good performance of the system when directly transferring simulation results into practical systems. Third, the optimal parameters for all discrete initial states are first learned offline and then tuned online to improve the generalization and real-time performance. In addition to the optimization control for the mobile robot, the proposed method also possesses general applicability for a class of nonlinear systems such as crane systems. Simulation and experimental results are included and analyzed to illustrate the superior performance of the proposed algorithm.

The tacrolimus-induced glucose homeostasis imbalance in terms of the liver: From bench to bedside.

Tacrolimus (TAC), the mainstay of maintenance immunosuppressive agents, plays a crucial role in new-onset diabetes after transplant (NODAT). Previous studies investigating the diabetogenic effects of TAC have focused on the ß cells of islets. In this study, we found that TAC contributed to NODAT through directly affecting hepatic metabolic homeostasis. In mice, TAC-induced hypoglycemia rather than hyperglycemia during starvation via suppressing gluconeogenetic genes, suggesting the limitation of fasting blood glucose in the diagnosis of NODAT. In addition, TAC caused hepatic insulin resistance and triglyceride accumulation through insulin receptor substrate (IRS)2/AKT and sterol regulatory element binding protein (SREBP1) signaling, respectively. Furthermore, we found a pivotal role of CREB-regulated transcription coactivator 2 (CRTC2) in TAC-induced metabolic disorders. The restoration of hepatic CRTC2 alleviated the metabolic disorders through its downstream molecules (eg, PCK1, IRS2, and SREBP1). Consistent with the findings from bench, low CRTC2 expression in graft hepatocytes was an independent risk factor for NODAT (odds ratio = 2.692, P = .023, n = 135). Integrating grafts' CRTC2 score into the clinical model could significantly increase the predictive capacity (areas under the receiver operating characteristic curve: 0.71 vs 0.79, P = .048). Taken together, in addition to its impact on pancreatic cells, TAC induces "hematogenous diabetes" via CRTC2 signaling. Liver-targeted management may be of help to prevent or heal TAC-associated diabetes.

Modulating interfacial charge distribution and compatibility boosts high energy density and discharge efficiency of polymer nanocomposites.

Polymer nanocomposite dielectrics, composed of polymer matrices with high breakdown strength and nanofillers with high dielectric constant, can achieve outstanding energy density. However, the great difference of intrinsic surface properties between the polymer and nanofillers will lead to poor compatibility and thus damage the dielectric properties of the composites. Introducing a transition layer to the filler surface can effectively reduce the degree of mismatch. In this work, we use a "direct in situ polymerization" method to synthesize core-shell BaTiO3 nanoparticles (BTO_nps) with three types of stable and dense fluoro-polymer shells, e.g., poly(2,2,2-trifluoroethyl methacrylate) (PTFEMA), poly(2,2,3,4,4,4-hexafluorobutyl methacrylate) (PHFBMA), and poly(1H,1H,7H-dodecafluoroheptyl methacrylate) (PDFHMA), and individually disperse them into the poly(vinylidene fluoride-co-hexafluoro propylene) (P(VDF-HFP)) matrix. Benefitting from the good interaction between the fluorine-containing segments in the shell polymer and the matrix segments, the dispersion of core-shell BTO_nps and their compatibility with P(VDF-HFP) are improved, which leads to a significant improvement in the dielectric properties of the nanocomposites. The results show that BTO@PDFHMA/P(VDF-HFP) composite exhibits an ultrahigh energy density of 16.8 J cm-3 at 609 MV m-1 with particle loading amount of 15 wt%, compared to 11.5 J cm-3 at 492 MV m-1 for a conventional solution blended BTO/P(VDF-HFP) composite. Meanwhile, the discharge efficiency is enhanced from ∼62 to ∼78%. It is elucidated that the core-shell strategy can achieve improved particle dispersion and dielectric properties. We consider that this simple method can well achieve the preparation of core-shell structures in dielectric nanocomposites.

Identification of two novel chicken GPR133 variants and their expression in different tissues.

GPR133 (G protein-coupled receptor 133) plays significant roles in various physiological processes. Alternatively splicing (AS) variants of GPR133 in many species have been predicted in multiple databases, but there is no available information about the AS events of chicken GPR133 (cGPR133). In the present study, two chicken GPR133 variants, cGPR133-va and cGPR133-vb, were identified by a combination of reverse transcription PCR (RT-PCR) and rapid amplification of cDNA 5'-ends (5' RACE). Sequence analysis shows that cGPR133-va and cGPR133-vb are resulting from different AS modules and their sequences are predicted to encode two distinct putative proteins, respectively. Quantitative real-time PCR (qRT-PCR) analysis revealed that cGPR133-va and cGPR133-vb are widely expressed in different tissues, while exhibiting specific expression profile. Altogether, our results first demonstrate the existence of novel cGPR133 variants and illustrate its transcriptional diversity and their widespread distribution, which provides a foundation for the further research of GPR133.

Identification of Three Novel Splicing Variants and Expression Analysis of Chicken GPR1 Gene.

GPR1 is a G protein-coupled receptor that plays critical roles in eukaryotic cells: typically, response to glucose stimulation, lipid accumulation, and transmitting nutrition signals to cAMP pathway. However, the alternative splicing of the GPR1 gene and its expression pattern in chicken tissues and ovarian follicles were unknown. In our current study, we used RACE-PCR to identify three GPR1 variants, including the full-length variant (GPR1-va1) and two alternatively spliced variants (GPR1-va2, GPR1-vb). Quantitative real-time PCR examined the expression pattern of GPR1 mRNA in chicken tissues and ovarian follicles. The result reveals that the coding sequence of the three variants cDNA is 1053, 1053, and 627 bp in length, encoding 350, 350, and 208 amino acids, respectively. The three variants of GPR1 show similar tissue distributions; GPR1 expression was abundant in the abdominal fat, lung, and heart. With the follicular development, the expression of GPR1 gene gradually increased, and GPR1-va1 and GPR1-va2 spliced variants expression in F2 were significantly higher than in F5, F4, and prehierarchical follicles (P < 0.05). Taken together, we found three novel variants of GPR1, and the results of GPR1 expression profiling in adipose tissues and ovarian follicles suggest that GPR1 may play a significant role in the lipid accumulation and progression of follicular development.