Comprehensive assessment of ocular parameters for identifying diagnostic indicators of diabetic peripheral neuropathy.

PURPOSE: To explore variations in systemic and ocular parameters among patients with diabetes, both with and without diabetic peripheral neuropathy (DPN) and to identify sensitive indicators for DPN diagnosis. METHODS: Ninty-five patients with type 2 diabetes mellitus (T2DM) were involved in this cross-sectional study, including 49 without DPN and 46 with DPN. Ocular parameters were obtained using optical coherence tomography angiography (OCTA) and corneal confocal microscopy (CCM). RESULT: Patients with DPN presented with significantly higher HbA1c (p < 0.05) and glycated albumin (GA, p < 0.01) levels, increased prevalence of diabetic retinopathy (DR, p < 0.05), and lower serum albumin (ALB, p < 0.01) and red blood cell (RBC, p < 0.05) levels. Ocular assessments revealed reduced corneal nerve fiber length (CNFL, p < 0.001) and enlarged foveal avascular zone (FAZ) area (p < 0.05) in DPN group. Logistic regression analysis indicated a significant association of presence of DR, RBC, GA, ALB, CNFL and DPN (p < 0.05, respectively). In the binary logistic regression for DPN risk, all three models including the presence of DR and CNFL exhibited the area under the curve (AUC) exceeding 0.8. CONCLUSION: The study establishes a strong correlation between ocular parameters and DPN, highlighting CCM's role in early diagnosis. Combining systemic and ocular indicators improves DPN risk assessment and early management.

Correction: Lu et al. Performance Analysis of Metalenses Based on Three Kinds of Phase Compensation Techniques. Nanomaterials 2021, 11, 2091.

There was an error in the original publication [...].

Effects of UV-B radiation on epiphytic bacterial communities on male and female Sargassum thunbergii.

The effects of increased UV-B radiation on macroalgae have been widely studied, but knowledge concerning the response of communities of algal epiphytic bacteria to increased UV-B radiation and differences between male and female algae is still lacking. Via 16S rDNA high-throughput sequencing technology, changes in the epiphytic bacterial communities on male and female S. thunbergii under increased UV-B radiation were studied in the lab. Under different UV-B radiation intensities, although the α diversity and community composition of epiphytic bacteria changed little, the ß diversity indicated that the community structure of bacteria on S. thunbergii was obviously clustered, and the relative abundance of dominant bacteria and indicator species changed considerably. There were unique bacteria in each experimental group, and the bacteria whose abundance obviously changed were members of groups related to environmental resistance or adaptability. The variation in the abundance of epiphytic bacteria was different in male and female S. thunbergii, and the bacteria whose abundance greatly changed were mainly related to algal growth and metabolism. The abundance of genes with predicted functions related to metabolism, genetic information processing, environmental adaptation and infectious diseases changed with increased UV-B radiation, and those variations differed between epiphytic bacteria on male and female S. thunbergii. This study found that the algal epiphytic bacteria were influenced by the increase in UV-B radiation and underwent certain adaptations through adjustments to community structure and function, and this response was also affected by the sex of the macroalgae. These results are expected to serve as experimental basis and provide reference for further understanding of the response of algae epiphytic bacteria to enhanced UV-B radiation caused by the thinning of the ozone layer and the resulting changes in the relationship between algae and bacteria, which may change the community of the marine ecosystem and affect important marine ecological process.

In vitro digestion and colonic fermentation of phenolic compounds and their bioaccessibility from raw and roasted nut kernels.

Roasting and digestion affect nut kernel phenolic compounds' bioaccessibility and bioactivity. In this study, three types of raw and commercially roasted nut kernels (almonds, cashews, and walnuts) were treated by in vitro digestion and colonic fermentation. The objective was to analyze the effect of roasting on their phenolic content, associated antioxidant potential, bioaccessibility, and short chain fatty acid (SCFA) synthesis altering. Among these, raw and roasted walnuts performed best, with significantly higher total phenolic content (TPC), total flavonoid content (TFC), free radical scavenging (2,2'-diphenyl-1-picrylhydrazyl (DPPH) assay) values, and ferric reducing antioxidant power (FRAP) values after completing gastrointestinal digestion. With the exception of cashews, roasting had no significant effect on antioxidant capacity during digestion from oral to small intestinal phase. Almonds showed the highest DPPH values after 16-hour colonic fermentation, reaching above 7.60 mg TE per g. Roasting had a positive effect on the free radical savagery capacity of walnuts within 16-24 hours of fecal fermentation. Significant differences were found in the bioaccessibility of individual compounds in raw and roasted nuts. As for almond and walnut, roasting increases the release and breakdown of phenolic compounds during colonic fermentation and have a positive impact on the bioaccessibility of specific phenolic compounds. The colonic bioaccessibility of most phenolic compounds was the highest. Due to heat polysaccharide breakdown, the total SCFAs produced were limited up to 0.03 mM. Raw almonds produced the most SCFAs at 16-hour fermentation and illustrated more benefits to gut health.

Multimodal imaging analysis for the impact of retinal peripheral lesions on central neurovascular structure and retinal function in type 2 diabetes with diabetic retinopathy.

OBJECTIVES: To explore the possible role of peripheral lesions (PLs) detected by ultrawide field (UWF) imaging system on central neurovascular structure and retinal function. METHODS: Ninety-seven diabetic patients were included in this cross-sectional study using UWF pseudocolour colour imaging with Optos Daytona (Optos, PLC). UWF images were graded as with predominantly peripheral lesions (PPLs) and without PPL. Macular neurovascular alterations and retinal function were measured by optical coherence tomography angiography (OCTA) and RETeval device, respectively. Central microcirculation and retinal function were compared between eyes with and without PPL. RESULTS: The study evaluated 186 eyes (97 patients; 43 females (44.3%)), including 92 eyes without PPL and 94 eyes with PPL. Central retinal vessel density was comparable between eyes with and without PPL. Delayed implicit time and decreased pupil area ratio were found in the PPL group compared with eyes without PPL, and this difference remained unchanged after adjusting for systemic factors (all p＜0.01). CONCLUSIONS: Our study suggests that retinal function is worse in diabetic eyes with PPL. These findings challenged the conventional ETDRS protocols which ignored peripheral retina in determining DR severity. Furthermore, combining UWF imaging with RETeval system to detect more retinal abnormalities may be helpful in DR management.

Versatile Integrated Polarizers Based on Geometric Metasurfaces.

We propose versatile integrated polarizers based on geometric metasurfaces. Metasurface polarizer consists of an L-shaped hole array etched on a silver film, and it can simultaneously generate several polarization states, including linear polarization, circular polarization, elliptical polarization, or even hybrid polarization. Meanwhile, the combination of output polarization states changes with the illumination polarization type. The theoretical analysis provides a detailed explanation for the generation of the integrated polarization states. The well-designed metasurface polarizers may generate more complex polarization modes, including vector beams and vector vortex beams. The theoretical and simulated results confirm the polarization performance of the proposed integrated metasurface polarizers. The compact design of metasurface polarizers and the controllable generation of versatile polarization combinations are a benefit to the applications of polarization light in optical imaging, biomedical sensing, and material processing.

Diabetic Retinopathy May Be a Predictor of Stroke in Patients With Diabetes Mellitus.

Background: It is unclear whether diabetic retinopathy (DR) can be a predictor of stroke. In this research context, the objective of our study was to investigate whether there is a significant association between DR and stroke in diabetic patients by meta-analysis. Methods: After a systematic search of studies in electronic databases, we screened all studies reporting the risk of DR status and stroke incidence and calculated their odds ratios (ORs) and hazard ratios (HRs). The effects of type of diabetes and severity of DR were also considered for subgroup analysis. Results: We included 19 studies involving 45 495 patients. A pooled HR = 1.62 (1.28-2.06) were found for the risk of DR and stroke in diabetic patients. In a subgroup analysis performed on the type of diabetes, the results showed a significant association between stroke incidence and DR status in patients with type 2 diabetes (T2D) (OR: 1.78; 95% CI, 1.53-2.08), but this association was not conclusive in type 1 diabetes (T1D) (OR: 1.77; 95% CI, 0.48-6.61). The results of the subgroup analysis with diabetes severity showed that both mild and moderate nonproliferative diabetic retinopathy (NPDR) status and severe NPDR and worse status significantly increased the risk of stroke with HRs of 2.01 (1.45-2.78) and 2.27 (1.52-3.39), respectively. Conclusion: DR status in diabetic patients is associated with an increased risk of stroke. This correlation was robust in patients with T2D, but uncertain in T1D. Based on this result, we have perhaps found the new factor for stroke management, so we analyzed the necessity and advantages of considering DR as a factor for stroke screening and risk management in our studies.

Impact of roasting on the phenolic and volatile compounds in coffee beans.

Phenolic compounds present in coffee beans could generate flavor and bring benefits to health. This study aimed to evaluate the impacts of commercial roasting levels (light, medium, and dark) on phenolic content and antioxidant potential of Arabica coffee beans (Coffea arabica) comprehensively via antioxidant assays. The phenolic compounds in roasted samples were characterized via liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS). Furthermore, the coffee volatile compounds were identified and semi-quantified by headspace/gas chromatography-mass spectrometry (HS-SPME-GC-MS). Generally, for phenolic and antioxidant potential estimation, light roasted samples exhibited the highest TPC (free: 23.97 ± 0.60 mg GAE/g; bound: 19.32 ± 1.29 mg GAE/g), DPPH, and FRAP. The medium roasted beans performed the second high in all assays but the highest ABTS+ radicals scavenging capacity (free: 102.37 ± 8.10 mg TE/g; bound: 69.51 ± 4.20 mg TE/g). Totally, 23 phenolic compounds were tentatively characterized through LC-ESI-QTOF-MS/MS, which is mainly adopted by 15 phenolic acid and 5 other polyphenols. The majority of phenolic compounds were detected in the medium roasted samples, followed by the light. Regarding GC-MS, a total of 20 volatile compounds were identified and semi-quantified which exhibited the highest in the dark followed by the medium. Overall, this study confirmed that phenolic compounds in coffee beans would be reduced with intensive roasting, whereas their antioxidant capacity could be maintained or improved. Commercial medium roasted coffee beans exhibit relatively better nutritional value and organoleptic properties. Our results could narrow down previous conflicts and be practical evidence for coffee manufacturing in food industries.

Physiological Responses and Metabonomics Analysis of Male and Female Sargassum thunbergii Macroalgae Exposed to Ultraviolet-B Stress.

Ultraviolet-B (UV-B) radiation is a major environmental stress that suppresses or activates defense responses in organisms. UV-B radiation affecting growth and development in intertidal species have been researched for a long time, but a series of unknown knowledge remain in the male and female macroalgae comparison. To compare the different responses of male and female Sargassum thunbergii macroalgae under UV-B radiation, PSII photochemical efficiency determination, metabolomic analysis, and main carbon-based metabolites (including soluble sugar, total amino acid, and lipid) content measuring have been performed in our experiments. Results showed that males have significantly superiority performance in the chlorophyll fluorescence parameters of F v/F m, Y(II), and Y(NO) either low or high UV-B radiation treatments. Metabolomics analysis revealed that carbon and nitrogen metabolism pathways in male and female S. thunbergii were significant components responding to enhanced UV-B radiation. Based on measuring, female S. thunbergii lipid content expressed higher than males without any stimulation. Additionally, under low UV-B radiation stimulation, females total amino acid content shown significantly higher than control group and their lipid content also significantly higher than males. Under high UV-B radiation, males soluble sugar, total amino acid, and lipid content significantly varied from females, which meant that enhancing UV-B stress might altered mainly carbon-based metabolites flowing directions. The present study elucidated the potential role of enhanced UV-B radiation in regulating macroalgae physiological responses, metabolites changing, and reflecting differences between male and female S. thunbergii, contributing to understanding of brown-macroalgae diecious adopting mechanisms in defending intertidal UV-B stresses.

Bioaccessibility and bioactivities of phenolic compounds from roasted coffee beans during in vitro digestion and colonic fermentation.

Bioaccessibility and bioactivity of phenolic compounds in coffee beans relate to roasting and digestion process. This study aimed to estimate phenolic content, antioxidant potential, bioaccessibility, and changes in short chain fatty acids (SCFAs) production during in vitro digestion and colonic fermentation of commercial roasted (light, medium and dark) coffee beans. There was no significant difference found among all three different roasting levels. TPC and DPPH were enhanced 15 mg GAE/g and 60 mg TE/g during gastrointestinal digestion, respectively. For colonic fermentation, the highest TPC and FRAP of all coffee beans was found at 2 and 4 h, respectively. The gastric bioaccessibility of most of the phenolic compounds were relatively higher due to thermal phenolic degradation. Total SCFAs production was only up to 0.02 mM because of thermal polysaccharide decomposition. Light roasted beans exhibited relatively higher phenolic bioaccessibility, antioxidant activities and SCFAs production, which would be more beneficial to gut health.

Current Insights into Phytochemistry, Nutritional, and Pharmacological Properties of Prosopis Plants.

Prosopis is a regional cash crop that is widely grown in arid, semiarid, tropical, and subtropical areas. Compared with other legume plants, Prosopis is underutilized and has great potentialities. Prosopis not only is a good source of timber, construction, fencing material, and gum, but also can be applied for food, beverage, feed, and medicine. Prosopis contains numerous phytochemical constituents, including carbohydrates, proteins, fatty acids, minerals, and vitamins, while varieties of phenolic compounds have also been identified from different parts of Prosopis. Flavonoids (especially C-glycosyl flavonoids), tannins, catechin, 4'-O-methyl-gallocatechin, mesquitol, and quercetin O-glycosides are significant phenolic contents in Prosopis. Various extracts of Prosopis displayed a wide range of biological properties, such as antioxidant, antihyperglycemic, antibacterial, anthelmintic, antitumor, and anticancer. Additionally, Prosopis has the potential to be an ideal diet that contains abundant dietary fiber, minerals, galactomannans, and low-fat content. However, the bioactivity and pharmacological properties associated with Prosopis were influenced by the bioavailability of phytochemicals, various antinutritional compounds, and the interactions of protein and phenolic compounds. The bioavailability of Prosopis is mainly affected by phenolic contents, especially catechin. The antinutritional compounds negatively affect the nutritional qualities of Prosopis, which can be prevented by heating. The protein-phenolic compound interactions can help the human body to absorb quercetin from Prosopis. This literature review aimed to provide systematic information on the physical, biochemical, pharmacological, and nutritional properties and potential applications of Prosopis.

Performance Analysis of Metalenses Based on Three Kinds of Phase Compensation Techniques.

The phase delays introduced by anisotropic nanounits include propagation phase delay, resonant phase delay and geometric phase delay. Various phase devices can be formed based on the metasurfaces consisting of anisotropic nanounits and the phase devices of the same kind function have different performances because of different working modes. In this paper, metalenses and vortex metalenses are chosen as examples to compare the optical performance of metasurface phase devices based on three kinds of phase compensation techniques. We design separately three kinds of metalenses and vortex metalenses using the cross nanoholes, L-shaped nanohole and V-shaped nanoholes and simulate numerically their intensity and phase distributions. Additionally, the results show the differences among these elements in structure complexity, polarization dependence, working efficiency and phase uniformity. The comparison for three kinds of metalenses clearly shows the merits of different phase compensation techniques and this work must be helpful for expanding the practical applications of metasurfaces.

Comparing the measurement properties of the EQ-5D-5L and the EQ-5D-3L in hypertensive patients living in rural China.

PURPOSE: The purpose of this study was to compare the measurement properties of two versions of EQ-5D (i.e.EQ-5D-3L and EQ-5D-5L) in hypertensive patients in rural China. METHODS: A cross-sectional survey was carried out in hypertensive patients in rural China. We compared the ceiling effects, redistribution properties, informativity, known-groups validity, and relative efficiency of the 3L and 5L and examined their agreement. RESULTS: A total of 11,412 patients were enrolled in our study. The mean EQ-5D index score was 0.84 (SD 0.21) according to the 5L and 0.86 (SD 0.17) according to the 3L. A good agreement was observed between the 3L and 5L. The overall ceiling effect decreased from 46.4% (3L) to 29.4% (5L). The Shannon index, H' improved in all dimensions when used 5L. When used 3L, the median responses of all groups were consistent with 5L across the three dimensions of 'mobility', 'self-care', 'usual activities', while the median responses were inconsistent for the 'pain/discomfort' and 'anxiety/depression' dimensions. The 3L performed better in eight comorbidities in terms of F-statistics and six comorbidities in terms of the area under the receiver operating characteristic curves (AUROCs). The 5L performed better both in terms of the F-statistics and AUROCs in age, education level, anti-hypertensive medication use. CONCLUSION: Taking all comparisons into account, we recommend the EQ-5D-5L for use in patients with hypertension in rural China.

Cost-effectiveness analysis of nivolumab plus ipilimumab versus chemotherapy as first-line treatment in advanced NSCLC.

Aim: To evaluate the cost-effectiveness of nivolumab plus ipilimumab (NI) in the first-line treatment of patients with advanced non-small-cell lung cancer from a US-payer perspective. Materials & methods: We developed a Markov model to evaluate the cost and effectiveness of NI versus chemotherapy as first-line treatment of NSCLC. Quality-adjusted life-years (QALYs) and incremental cost-effectiveness ratios (ICERs) were estimated. Results: NI provided an additional 0.715 QALYs compared with chemotherapy in all population. The corresponding ICER of NI was $180,307 per QALY gained. However, the ICER decreased to $143,434 per QALY in the programmed death ligand 1 expression level <1% population. Conclusion: From a US-payer perspective, NI is estimated to be cost-effective in the first-line setting for advanced NSCLC patients with programmed death ligand 1 expression level <1%.

Cost-effectiveness analysis of atezolizumab plus nab-paclitaxel for untreated metastatic triple-negative breast cancer.

Aim: To evaluate the cost-effectiveness of atezolizumab plus nab-paclitaxel (ANP) in the first-line treatment of metastatic triple-negative breast cancer (TNBC). Materials & methods: We developed a Markov model to evaluate the cost and effectiveness of ANP versus nab-paclitaxel in the first-line treatment of metastatic TNBC. Lifetime costs, life-years (LYs) and quality-adjusted LYs (QALYs) were estimated. Results: ANP provided an additional 0.16 QALYs (0.24 LYs) compared with nab-paclitaxel in intention-to-treat population. The corresponding incremental cost-effectiveness ratio was $786,131 per QALY gained. However, the incremental cost-effectiveness ratio decreased to $361,218 per QALY gained in the PD-L1 positive subgroup analysis. Conclusion: From the perspective of a US-payer, ANP is estimated not to be cost-effective in the first-line treatment of metastatic TNBC.

A Cost-Effectiveness Analysis: First-Line Avelumab Plus Axitinib Versus Sunitinib for Advanced Renal-Cell Carcinoma.

BACKGROUND: Compared with the standard of care with sunitinib, avelumab plus axitinib can increase progression-free survival in the first-line of advanced renal cell carcinoma (RCC), but the economic effect of the treatment is unknown. The purpose of the research was to evaluate the cost-effectiveness of the avelumab plus axitinib versus sunitinib in first-line treatment for advanced RCC from the US payer perspective. METHODS: A Markov model was developed to evaluate the economic and health outcomes of avelumab plus axitinib vs sunitinib in the first-line setting for advanced RCC. The clinical data were obtained from the JAVELIN Renal 101 Clinical Trials. Deterministic and probabilistic sensitivity analyses were performed to assess uncertainty in the model. Health outcomes were measured in quality-adjusted life-years (QALYs). RESULTS: The incremental cost-effectiveness ratio (ICER) of avelumab plus axitinib compared with sunitinib was $565,232 per QALY, the costs were $884,626 and $669,838, QALYs were 3.67 and 3.29, respectively. Sensitivity analysis demonstrated that differences in utilities in PFS and after progression were the most influential factors within the model. When avelumab was at 30% of the full price or axitinib was at 40% of the full price, avelumab and axitinib were approved to be cost-effective if the WTP threshold was $150,000 per QALY. The subgroup analysis showed the ICER of avelumab plus axitinib compared with sunitinib for the patients with PD-L1-positive tumors was $588,105. CONCLUSION: Avelumab plus axitinib in the first-line treatment was not cost-effective in comparison with sunitinib when the threshold of willingness to pay (WTP) was $150,000 per QALY.

Identification of DNA methylation-driven genes by integrative analysis of DNA methylation and transcriptome data in pancreatic adenocarcinoma.

Pancreatic adenocarcinoma (PAAD) is a painful and fatal disease that undoubtedly remains a health care priority and offers significant therapeutic challenges. The significance of epigenetic modifications, including DNA methylation in tumor development, has gained the attention of researchers. Identifying DNA methylation-driven genes and investigating the mechanisms underlying the tumorigenesis of PAAD are of substantial importance for developing methods of physiological evaluation, treatment planning and prognostic prediction for PAAD. In the present study, a comprehensive analysis of DNA methylation and gene expression data from 188 clinical samples was performed to identify DNA methylation-driven genes in PAAD. In addition, the diagnostic and prognostic value of DNA methylation-driven genes was evaluated using receiver operating characteristic curve, survival and recurrence analyses. A total of 7 DNA methylation-driven genes, namely zinc finger protein 208 (ZNF208), eomesodermin (EOMES), prostaglandin D2 receptor (PTGDR), chromosome 12 open reading frame 42 (C12orf42), integrin subunit α 4 (ITGA4), dedicator of cytokinesis 8 and protein phosphatase 1 regulatory inhibitor subunit 14D (PPP1R14D), were identified. All of them may be used to diagnose PAAD with excellent specificity and sensitivity (area under curve, >0.8). Of the 7 DNA methylation-driven genes, 6 were significantly associated with overall survival (OS) and recurrence-free survival (RFS) P<0.05). Among them, ZNF208, EOMES, PTGDR, C12orf42 and ITGA4 were significantly negatively associated with the OS rate and positively associated with the recurrence rate, while PPP1R14D was significantly positively associated with the OS rate and negatively associated with the recurrence rate. The present study provides novel insight into the epigenetic alterations associated with the occurrence and progression of PAAD, thereby increasing the mechanistic understanding of this disease, offering potential novel molecular biomarkers and contributing to the development of therapeutic targets for PAAD.

Standardized ileal digestible amino acid and metabolizable energy content of wheat from different origins and the effect of exogenous xylanase on their determination in broilers.

This study was conducted to determine the standardized ileal digestible amino acids (SID AA) and nitrogen-corrected apparent metabolizable energy (AMEn) contents of 6 wheats from different origins in China and incidentally to investigate the effects of exogenous xylanase addition on SID AA and AMEn determination in broiler chicks. A total of 480 chicks were divided into 48 cages of 10 birds each balanced for body weight and fed 8 types of diets in a completely randomized design (6 replicated cages per diet) from 21 to 26 d of age. The individual wheat constituted the only source of crude protein in a semi-purified experimental diet. A nitrogen-free diet was designed to estimate basal endogenous AA loss and determine the SID AA. Titanium oxide (0.3%) was used as an indigestibility marker, and nutrient digestibility and retention were determined by the substitution method. From day 24 to 26, excreta samples were collected for AMEn determination. On day 26, the birds were euthanized, and ileum contents were obtained for AA digestibility determination. Wheat from Gansu had greater (P < 0.05) SID AA contents except Lys, Thr, Phe, and Cys, with a higher (P < 0.001) AMEn (11.83 MJ/kg) than the other wheats. The SID content of mean indispensable amino acids and dispensable amino acids were 87.35% and 88.17%, respectively, and the average AMEn value of 6 wheats was 11.14 MJ/kg. Compared with the diet without xylanase, the added xylanase resulted in higher (P < 0.05) SID contents of Met, Lys, Trp, Arg, Ile, Leu, Val, Gly, Asp, Glu, Pro, and Ala; the SID AA values were raised by 1.96% (mean of all AA); and the AMEn content was significantly increased (+0.87 MJ/kg) (P < 0.05). In conclusion, origins of wheats have significant effects on SID AA and AMEn values which were positively correlated with crude protein content of wheat; exogenous xylanase addition to a wheat-based poultry diet could significantly improve SID AA and AMEn contents for broilers.

A Novel PPh3 Mediated One-Pot Method for Synthesis of 3-Aryl or Alkyl 1,2-Benzisoxazoles.

A novel, efficient, and facile protocol has been developed for transforming 2-hydroxybenzonitriles and bromides into a range of 3-aryl or alkyl substituted 1,2-benzisoxazoles in good to excellent yields mediated by PPh3. The electronic and steric effects of bromides on the reaction are discussed. This is the first example to construct a C-C bond and heterocycle in a Barbier-Grignard-type reaction featuring easier recovery of PPh3 than a metallic catalyst in one step.

Relative abundance of G protein-coupled receptor 30 and localization in testis and epididymis of sheep at different developmental stages.

The G protein-coupled receptor 30 (GPR30) is a transmembrane estrogen receptor that binds to estrogen, and has been confirmed to have an important role in testicular cell proliferation and development. The main objective of this study was to examine GPR30 gene expression and localization in the testis and epididymis of sheep at different developmental stages. Testes, including the epididymis, were collected from Polled Dorset x Mongolian cross rams at one (n=4; wt), three (n=4; wt), six (n=4; wt), nine (n=4; wt) and 12 (n=4; wt) months of age. The 12-month-old hybrid crossbred sheep were exsanguinated via puncture of the jugular vein. Relative abundance of GPR30 mRNA was detected by quantitative PCR, and localization of the protein was examined by immunohistochemistry. Semi-quantitative analysis of GPR30 protein was performed by western blotting. The relative abundance of GPR30 mRNA was similar in the epididymis tail for rams at 6, 9, and 12mo of age. Further, relative abundance of GPR30 mRNA in the testes and caput epididymis of 1-, 3-, 6-, 9-, and 12-month-old crossbred rams did not increase with age. The GPR30 mRNA was detected in epididymal interstitial and principal cells, and in the epididymal cavity, spermatocytes, spermatogonial stem cells, Sertoli and Leydig cells, and spermatozoon of ram testes. Western blotting indicated the GPR30 protein was present in 9- and 12-month-old crossbred sheep corpus, cauda epididymis (P<0.05). The results suggest that relative abundance of GPR30 mRNA is time-dependent and localization-specific.