Detection and Analysis of Genes Affecting the Number of Thoracic Vertebrae in Licha Black Pigs.

The number of thoracic vertebrae (NTV) in pigs is an important economic trait that significantly influences pork production. While the Licha black pig is a well-known Chinese pig breed with multiple thoracic vertebrae, the genetic mechanism is still unknown. Here, we performed a selective signal analysis on the genome of Licha black pigs, comparing individuals with 15 NTV versus those with 16 NTV to better understand functional genes associated with NTV. A total of 2265 selection signal regions were detected across the genome, including 527 genes and 1073 QTL that overlapped with the selection signal regions. Functional enrichment analysis revealed that LRP5 and SP5 genes were involved in biological processes such as bone morphogenesis and Wnt protein binding. Furthermore, three genes, LRP8, DEF6, and SCUBE3, associated with osteoblast differentiation and bone formation, were located within or close to the QTL related to bone development and vertebrae number. These five genes were hypothesized to be potential candidates for regulating the NTV trait in Licha black pigs. Our findings revealed several candidate genes that play crucial roles in NTV regulation and provide a theoretical foundation to understand the genetic mechanism of the NTV trait in pig breeding.

Enhanced morphological assessment based on interocular asymmetry analysis for keratoconus detection.

PURPOSE: To clarify the interocular asymmetry of corneal morphological descriptors and evaluate its discriminant ability of keratoconus (KC). METHODS: This retrospective study recruited 344 normal participants and 290 KC patients, randomized to training and validation datasets. Interocular correlation and agreement were evaluated on 44 corneal morphological descriptors derived from Schiempflug tomography. Logistic regression models were constructed using binocular data and of which diagnostic performance was evaluated using the area under receiver operating characteristics curve (AUC), net reclassification improvement (NRI), and integrated discrimination improvement (IDI). RESULTS: Interocular agreement of corneal descriptors is better in the normal than in KC except for dimensions of cornea and anterior chamber. The interocular asymmetry increases along with the severity of KC. Interocular asymmetry in maximum anterior keratometry, mean anterior keratometry and higher-order aberrations of anterior surface show high AUC above 0.950. Binocular logistic regression index reaches an AUC of 0.963 with high specificity (95.2%) and brings gain to monocular parameters in distinguishing the normal eyes from KC (NRI = 0.080 (0.042 ~ 0.118), P < 0.001) and IDI = 0.071 (0.049 ~ 0.092), P < 0.001). Interocular asymmetry benefits even more in subclinical keratoconus (SKC) detection reflected by NRI (0.4784 (0.2703-0.6865), P < 0.001) and IDI (0.2680 (0.1495-0.3866), P < 0.001) measures. CONCLUSION: Interocular asymmetry is a well-characterized feature of KC and related to the severity. It is feasible to apply the interocular asymmetry in diagnosis of KC and SKC as a replenishment of monocular parameters and in progression tracking.

The application of corneal biomechanical interocular asymmetry for the diagnosis of keratoconus and subclinical keratoconus.

Purpose: To evaluate the interocular consistency of biomechanical properties in normal, keratoconus (KC) and subclinical keratoconus (SKC) populations and explore the application of interocular asymmetry values in KC and SKC diagnoses. Methods: This was a retrospective chart-review study of 331 ametropic subjects (control group) and 207 KC patients (KC group, including 94 SKC patients). Interocular consistency was evaluated using the intraclass correlation coefficient (ICC). Interocular asymmetry was compared between the control and KC groups and its correlation with disease severity was analyzed. Three logistic models were constructed using biomechanical monocular parameters and interocular asymmetry values. The diagnostic ability of interocular asymmetry values and the newly established models were evaluated using receiver operating characteristic curves and calibration curves. Net reclassification improvement (NRI) and integrated discrimination improvement (IDI) were also estimated. Results: The interocular consistency significantly decreased and the interocular asymmetry values increased in KC patients compared with those in control individuals. In addition, the interocular asymmetry values increased with respect to the severity of KC. The binocular assisted biomechanical index (BaBI) had an area under the curve (AUC) of 0.998 (97.8% sensitivity, 99.2% specificity; cutoff 0.401), which was statistically higher than that of the Corvis biomechanical index [CBI; AUC = 0.935, p < 0.001 (DeLong's test), 85.6% sensitivity]. The optimized cutoff of 0.163 provided an AUC of 0.996 for SKC with 97.8% sensitivity, which was higher than that of CBI [AUC = 0.925, p < 0.001 (DeLong's test), 82.8% sensitivity]. Conclusion: Biomechanical interocular asymmetry values can reduce the false-negative rate and improve the performance in KC and SKC diagnoses.

Molecules related to diabetic retinopathy in the vitreous and involved pathways.

Diabetic retinopathy (DR) is one of the most common complications of diabetes and major cause of blindness among people over 50 years old. Current studies showed that the vascular endothelial growth factor (VEGF) played a central role in the pathogenesis of DR, and application of anti-VEGF has been widely acknowledged in treatment of DR targeting retinal neovascularization. However, anti-VEGF therapy has several limitations such as drug resistance. It is essential to develop new drugs for future clinical practice. The vitreous takes up 80% of the whole globe volume and is in direct contact with the retina, making it possible to explore the pathogenesis of DR by studying related factors in the vitreous. This article reviewed recent studies on DR-related factors in the vitreous, elaborating the VEGF upstream hypoxia-inducible factor (HIF) pathway and downstream pathways phosphatidylinositol diphosphate (PIP2), phosphoinositide-3-kinase (PI3K), and mitogen-activated protein kinase (MAPK) pathways. Moreover, factors other than VEGF contributing to the pathogenesis of DR in the vitreous were also summarized, which included factors in four major systems, kallikrein-kinin system such as bradykinin, plasma kallikrein, and coagulation factor XII, oxidative stress system such as lipid peroxide, and superoxide dismutase, inflammation-related factors such as interleukin-1ß/6/13/37, and interferon-γ, matrix metalloproteinase (MMP) system such as MMP-9/14. Additionally, we also introduced other DR-related factors such as adiponectin, certain specific amino acids, non-coding RNA and renin (pro) receptor in separate studies.

A prospective randomized self-controlled study of LASIK combined with accelerated cross-linking for high myopia in Chinese: 24-month follow-up.

BACKGROUND: To assess the visual and refractive outcomes of femtosecond laser-assisted in situ keratomileusis (FS-LASIK) concurrent with accelerated cross-linking (LASIK Xtra) compared with conventional FS-LASIK (convLASIK) for high myopia in Chinese individuals. METHODS: In this prospective, randomized, fellow-eye comparison study, 25 patients with high myopia were treated randomly with LASIK Xtra in one eye and convLASIK in the other. A 24-month follow-up was conducted, and the main outcome measures included uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), manifest refraction spherical equivalent (MRSE) and corneal tomography. RESULTS: The UDVA was 0.09 ± 0.15 logMAR in the LASIK Xtra group, which was significantly worse than that in the convLASIK group 1 day postoperatively (P = .001), but the difference became nonsignificant from 1 week after surgery. The efficacy index was 0.88 ± 0.18 in the LASIK Xtra eyes and 0.99 ± 0.13 in the convLASIK eyes at 24 months (P = .028). Throughout the follow-up period, a slight myopic shift in the MRSE and keratometry values were observed in both groups without significant intergroup differences. The coefficient of determination was 0.9982 in the LASIK Xtra eyes and 0.9987 in the convLASIK eyes. The corneal density was significantly higher, and demarcation lines were visible in the first 6 months in LASIK Xtra eyes, but both signs of cross-linking gradually disappeared during follow-up. No severe complications were detected in either group. CONCLUSIONS: LASIK Xtra showed comparable safety and predictability with convLASIK for high myopia in Chinese, but lower efficacy and no greater stability was observed up to 24-month follow-up.

Effects of riboflavin on boar sperm motility, sperm quality, enzyme activity and antioxidant status during cryopreservation.

OBJECTIVES: This study was conducted to evaluate the effect of adding riboflavin to boar sperm freezing extender on the challenge of cryopreservation. METHODS: Different concentrations (0, 5, 10, 15, 20 or 25 µM) of riboflavin were added to the freezing extender. Spermatozoa motility, membrane integrity, acrosomal integrity, mitochondrial membrane potential and enzyme activities were analysed once 10 min after thawing. Q-PCR was used to detect the mRNA expression of Caspase3, Bcl-2 and Bax. RESULTS: The results showed that the addition of 10 µM riboflavin to boar sperm freezing extender significantly increased the frozen-thawed sperm progressive motility compared with the control group (p < 0.05). Activities of superoxide dismutase, glutathione peroxidase and catalase improved after adding riboflavin to the extender (p < 0.05). During freezing-thawing, the boar sperm mitochondrial membrane potential, acrosomal integrity, plasma membrane and DNA at 10 µM in the riboflavin group increased by 6.6%, 9.6%, 5.49% and 5.62% (p < 0.05), respectively, compared with the control group. The addition of 10 µM riboflavin to the extender significantly decreased the malondialdehyde (p < 0.05) content, whereas it increased the ATP content (p < 0.05) of boar sperm during freezing-thawing. Furthermore, the expression of Caspase-3 and Bax (p < 0.05) were significantly lower, whereas the expression of BCL-2 (p < 0.05) was greater than the control group when adding 10 µM riboflavin to the extender. CONCLUSIONS: Riboflavin showed cryoprotective capacity to the freezing extender used for boar sperm during the process of freezing-thawing, and the optimal concentration of riboflavin for the frozen extender was 10 µM.

Analysis of the Genetic Diversity and Family Structure of the Licha Black Pig Population on Jiaodong Peninsula, Shandong Province, China.

The Licha black pig, a popular indigenous Chinese pig breed, is known for its multi-vertebral trait and higher lean meat rate. Understanding the current conservation status, family structure, and degree of inbreeding of the Licha black pig population will be useful to maintain a sufficient level of genetic diversity in these animal resources. In the present study, the genetic diversity, population structure, and inbreeding coefficient of this conserved population were analyzed using SNP genotyping data from 209 Licha black pigs. Based on the genomic information, this population was divided into eight different families with boars. The effective population size (Ne), polymorphic marker ratio (PN), expected heterozygosity (He), and observed heterozygosity (Ho) of this population were 8.7, 0.827, 0.3576, and 0.3512, respectively. In addition, a total of 5976 runs of homozygosity (ROHs) were identified, and most of the ROHs (54.9%) were greater than 5 Mb. The genomic inbreeding coefficient of each individual was estimated based on ROHs (FROH) with an average inbreeding coefficient of 0.11 for the population. Five statistics (Ne, PN, Ho, He, and FROH) showed a decrease in the level of genetic diversity and a high degree of inbreeding in this population. Thus, special preservation programs need to be implemented in the future, such as introducing new individuals or improving the mating plan. Altogether, our study provides the first genomic overview of the genetic diversity and population structure of Licha black pigs, which will be useful for the management and long-term preservation of this breed.

Visual functional defects in patients with type 2 diabetes mellitus: a questionnaire based cross-sectional study.

PURPOSE: To determine the impact of type 2 diabetes mellitus (T2DM) on visual functions, identify different modifiers as risk or protective factors, and find out how these factors affect patients' visual symptoms and visual functions as a whole. METHODS: We performed an online survey among 1030 participants (400 patients, 630 non-patients). Demographic features and severity of disease were documented, while visual functions were evaluated using National Eye Institute Visual Functioning questionnaire-25 (NEI VFQ-25). Independent t-test, analysis of variance, linear and nonlinear regression models were used to assess all data. RESULTS: Scores other than color vision among T2DM patients were significantly lower compared with non-T2DM participants. There was significant difference after stratification of age and education, but no significant difference between different genders was observed. Parameters including duration of T2DM, fasting plasma glucose (FPG) and glycosylated hemoglobin A1c (HbA1c) negatively impacted on the scores, with 20 years' of diabetic duration, 10 mmol/L of FPG, 7.5% of HbA1c being potential cut-off points. Poorer best corrected visual acuity (BCVA) and diagnosis of diabetic retinopathy were risk factors, while they simultaneously produced mediation effect, contributing 5%-78% of effect in the deterioration of visual functions caused by longer diabetic duration and higher blood glucose. CONCLUSION: Significant visual impairments and faster deterioration in visual functions were seen in T2DM patients, with older age, lower educational level, longer diabetic duration, poorer blood glucose administration, limited BCVA, and the presence of diabetic retinopathy identified as risk factors. Average BCVA and diabetic retinopathy also yielded mediation effect as diabetic duration lengthened and blood glucose elevated.

Estimating in vitro ruminal ammonia-N using multiple linear models and artificial neural networks based on the CNCPS nitrogenous fractions of cattle rations with low concentrate/roughage ratios.

The objectives of this study were to investigate the relationship between the in vitro ruminal ammonia nitrogen (NH3 -N) concentration and the Cornell Net Carbohydrate and Protein System (CNCPS) N-fractions of feeds for cattle and further compare the performance of developing multiple linear regression (MLR) and artificial neural network (ANN) models in estimating the NH3 -N concentration in rumen fermentation. Two data sets were established, of which the training data set containing forty-five rations for cattle with concentrate/roughage ratios of 50:50, 40:60, 30:70, 20:80 and 10:90 used for developing models and the test data set containing ten other rations with the same concentrate/roughage ratios with the training data set were used for validating of models. The NH3 -N concentrations of feed samples were measured using an in vitro incubation technique. The CNCPS N-fractions (g), for example PB1 (rapidly degraded true protein), PB2 (neutral detergent soluble nitrogen), PB3 (acid detergent soluble nitrogen) of rations, were calculated based on chemical analysis. Statistical analysis indicated that the NH3 -N concentration (mg) was significantly correlated with the CNCPS N-fractions (g) PB1 , PB2 and PB3 in a multiple linear pattern: NH3 -N = (130.70±33.80) PB1  + (155.83±17.89) PB2 - (85.44±37.69) PB3  + (42.43±1.05), R2  = 0.77, p < 0.0001, n = 45. The results indicated that both MLR and ANN models were suitable for predicting in vitro NH3 -N concentration of rations using CNCPS N-fractions PB1 , PB2 , and PB3 as independent variables while the neural network model showed better performance in terms of greater r2 , CCC and lower RMSPE between the observed and predicted values.

A Review of 3-Nitrooxypropanol for Enteric Methane Mitigation from Ruminant Livestock.

Methane (CH4) from enteric fermentation accounts for 3 to 5% of global anthropogenic greenhouse gas emissions, which contribute to climate change. Cost-effective strategies are needed to reduce feed energy losses as enteric CH4 while improving ruminant production efficiency. Mitigation strategies need to be environmentally friendly, easily adopted by producers and accepted by consumers. However, few sustainable CH4 mitigation approaches are available. Recent studies show that the chemically synthesized CH4 inhibitor 3-nitrooxypropanol is one of the most effective approaches for enteric CH4 abatement. 3-nitrooxypropanol specifically targets the methyl-coenzyme M reductase and inhibits the final catalytic step in methanogenesis in rumen archaea. Providing 3-nitrooxypropanol to dairy and beef cattle in research studies has consistently decreased enteric CH4 production by 30% on average, with reductions as high as 82% in some cases. Efficacy is positively related to 3-NOP dose and negatively affected by neutral detergent fiber concentration of the diet, with greater responses in dairy compared with beef cattle when compared at the same dose. This review collates the current literature on 3-nitrooxypropanol and examines the overall findings of meta-analyses and individual studies to provide a synthesis of science-based information on the use of 3-nitrooxypropanol for CH4 abatement. The intent is to help guide commercial adoption at the farm level in the future. There is a significant body of peer-reviewed scientific literature to indicate that 3-nitrooxypropanol is effective and safe when incorporated into total mixed rations, but further research is required to fully understand the long-term effects and the interactions with other CH4 mitigating compounds.

Evaluation and reflection on claudin 18.2 targeting therapy in advanced gastric cancer.

Claudin 18.2 (CLDN18.2) is a tight-junction protein. CLDN18.2-targeting strategy has cut a striking figure in CLDN18.2 positive patients with advanced gastric cancer. Zolbetuximab, the CLDN18.2 antibody, obtained a better clinical benefit in patients compared with the controlled. In phase II trials, combination treatment of epirubicin, oxaliplatin and capecitabine (EOX) + zolbetuximab achieved the optimal effects of overall survival which extended to 13.2 months with tolerable safety events, indicating its greater potential playing the second promising target in gastric cancer. This review will reveal the definitive clinical benefit CLDN18.2-targeting therapies have achieved and update the highlighting development (like chimeric antigen receptor T-cell immunotherapy) to CLDN18.2 positive patients. We then focus on 10 questions arisen from recent progress and anticipate to provide a future perspective for novel cancer treatment.

Predicting in vitro rumen VFA production using CNCPS carbohydrate fractions with multiple linear models and artificial neural networks.

The objectives of this trial were to develop multiple linear regression (MLR) models and three-layer Levenberg-Marquardt back propagation (BP3) neural network models using the Cornell Net Carbohydrate and Protein System (CNCPS) carbohydrate fractions as dietary variables for predicting in vitro rumen volatile fatty acid (VFA) production and further compare MLR and BP3 models. Two datasets were established for the trial, of which the first dataset containing 45 feed mixtures with concentrate/roughage ratios of 10∶90, 20∶80, 30∶70, 40∶60, and 50∶50 were used for establishing the models and the second dataset containing 10 feed mixtures with the same concentrate/roughage ratios with the first dataset were used for testing the models. The VFA production of feed samples was determined using an in vitro incubation technique. The CNCPS carbohydrate fractions (g), i.e. CA (sugars), CB1 (starch and pectin), CB2 (available cell wall) of feed samples were calculated based on chemical analysis. The performance of MLR models and BP3 models were compared using a paired t-test, the determination coefficient (R2) and the root mean square prediction error (RMSPE) between observed and predicted values. Statistical analysis indicated that VFA production (mmol) was significantly correlated with CNCPS carbohydrate fractions (g) CA, CB1, and CB2 in a multiple linear pattern. Compared with MLR models, BP3 models were more accurate in predicting acetate, propionate, and total VFA production while similar in predicting butyrate production. The trial indicated that both MLR and BP3 models were suitable for predicting in vitro rumen VFA production of feed mixtures using CNCPS carbohydrate fractions CA, CB1, and CB2 as input dietary variables while BP3 models showed greater accuracy for prediction.

Relationship between the Methane Production and the CNCPS Carbohydrate Fractions of Rations with Various Concentrate/roughage Ratios Evaluated Using In vitro Incubation Technique.

The objective of the trial was to study the relationship between the methane (CH4) production and the Cornell Net Carbohydrate and Protein System (CNCPS) carbohydrate fractions of feeds for cattle and the suitability of CNCPS carbohydrate fractions as the dietary variables in modeling the CH4 production in rumen fermentation. Forty-five rations for cattle with the concentrate/roughage ratios of 10:90, 20:80, 30:70, 40:60, and 50:50 were formulated as feed samples. The Menke and Steingass's gas test was used for the measurement of CH4 production. The feed samples were incubated for 48 h and the CH4 production was analyzed using gas chromatography. Statistical analysis indicated that the CH4 production (mL) was closely correlated with the CNCPS carbohydrate fractions (g), i.e. CA (sugars); CB1 (starch and pectin); CB2 (available cell wall) in a multiple linear pattern: CH4 = (89.16±14.93) CA+ (124.10±13.90) CB1+(30.58±11.72) CB2+(3.28±7.19), R(2) = 0.81, p<0.0001, n = 45. Validation of the model using 10 rations indicated that the CH4 production of the rations for cattle could accurately be predicted based on the CNCPS carbohydrate fractions. The trial indicated that the CNCPS carbohydrate fractions CA, CB1 and CB2 were suitable dietary variables for predicting the CH4 production in rumen fermentation in vitro.