Enhanced serum levels of tumor necrosis factor-α, interleukin-1ß, and -6 in sarcopenia: alleviation through exercise and nutrition intervention.

BACKGROUND: Limited research has been conducted on the post-intervention inflammatory status in sarcopenic patients, despite previous studies revealing elevated pro-inflammatory markers. This study aimed to investigate the potential elevation of specific pro-inflammatory cytokines in sarcopenic patients and evaluate the effects of exercise and nutritional support interventions on these cytokine levels. METHODS: In this post-hoc analysis of a randomized controlled trial (RCT), 57 individuals with sarcopenia from the RCT and 57 non-sarcopenic participants from the same geriatric community cohort that did not participate in the RCT were enrolled. Grip strength and body composition measurements were recorded. Tumor necrotizing factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-15 levels were assessed at baseline for both groups and after a 12-week intervention consisting of resistive exercise and supplementation with branched-chain amino acids, calcium, and vitamin D3 in the patients with sarcopenia. RESULTS: The sarcopenic group demonstrated significantly lower body weight, body mass index, grip strength, and skeletal muscle mass index. Moreover, sarcopenic patients exhibited higher levels of TNF-α (p=0.007), IL-1ß (p<0.001), and IL-6 (p<0.001), while no significant difference was observed in IL-15 (p=0.345) between participants with and those without sarcopenia. Following the intervention, the sarcopenic group experienced significant improvements in grip strength and skeletal muscle mass index with a notable reduction in TNF-α (p=0.003), IL-1ß (p=0.012) and IL-6 (p=0.001) levels. CONCLUSIONS: Sarcopenic patients exhibit elevated levels of TNF-α, IL-1ß, and IL-6, which declined after nutrition support and exercise interventions. However, further research is necessary to evaluate the long-term impact of these interventions on cytokine levels.

Assessing thickness and stiffness of superficial/deep masticatory muscles in orofacial pain: an ultrasound and shear wave elastography study.

INTRODUCTION: Sonoelastography has been increasingly used for non-invasive evaluation of the mechanical features of human tissues. The interplay between orofacial pain and regional muscle activity appears clinically paramount, although only few imaging studies have investigated this association. Using shear wave sonoelastography (SWS), this study ascertained whether orofacial pain induced alterations in the stiffness of superficial and deep masticatory muscles. METHODS: All participants were systematically evaluated for oral/facial-related conditions, including the area and intensity of pain. SWS was applied to measure the stiffness of the bilateral masseter, temporalis, and lateral pterygoid muscles. The association between orofacial pain and muscle stiffness/thickness was investigated using a generalized estimating equation for adjusting the influence of age, sex, laterality, and body mass index on muscle thickness/stiffness. RESULTS: A total of 98 participants were included in the present study: 48 asymptomatic controls, 13 patients with unilateral pain, and 37 patients with bilateral orofacial pain. The reliability, quantified by the intraclass correlation coefficient for muscle stiffness measurement, ranged from 0.745 to 0.893. Orofacial pain at the individual muscle level was significantly associated with masseter muscle stiffness. A trend of increased stiffness (p = 0.06) was also observed in relation to the painful side of the temporalis muscle. No significant correlation was identified between the numeric rating scales for pain and stiffness measurements. CONCLUSIONS: SWS provides reliable stiffness measurements for the superficial and deep masticatory muscles. The ipsilateral masseter and temporalis muscles might be stiffer than those on the side without orofacial pain. Future studies using the present sonoelasotography protocol can be designed to investigate the stiffness changes in the target muscles after interventions.

Shear wave sonoelastography (SWS) can reliably assess the stiffness of masticatory muscles.Orofacial pain, particularly affecting the ipsilateral masseter muscles, exhibited increased stiffness, with a similar trend observed in the temporalis muscle as revealed by SWS. However, the stiffness of the lateral pterygoid muscle appeared to remain unaffected.These findings establish a foundational framework for the objective and quantitative assessment of orofacial pain and indicate the potential utility of SWS as a tool for evaluating treatment outcomes.

[Effect of High-Concentration Uric Acid on Nitric Oxide].

Uric acid (UA) is the final product of purine metabolism in human body,and its metabolic disorder will induce hyperuricemia (HUA).The occurrence and development of HUA are associated with a variety of pathological mechanisms such as oxidative stress injury,activation of inflammatory cytokines,and activation of renin-angiotensin-aldosterone system.These mechanisms directly or indirectly affect the bioavailability of endogenous nitric oxide (NO).The decrease in NO bioavailability is common in the diseases with high concentration of UA as an independent risk factor.In this review,we summarize the mechanisms by which high concentrations of UA affect the endogenous NO bioavailability,with a focus on the mechanisms of high-concentration UA in decreasing the synthesis and/or increasing the consumption of NO.This review aims to provide references for alleviating the multisystem symptoms and improving the prognosis of HUA,and lay a theoretical foundation for in-depth study of the correlations between HUA and other metabolic diseases.

The impact of shoulder pathologies on job discontinuation and return to work: a pilot ultrasonographic investigation.

BACKGROUND: Ultrasound imaging has emerged as one of the most useful tools for evaluating shoulder disorders. To date, the association between shoulder ultrasonography and a patient's work status has rarely been explored by antecedent studies. AIM: This study aimed to investigate the association between sonographically diagnosed shoulder pathologies and job discontinuation and return to work. DESIGN: A cross-sectional study. SETTING: Outpatient clinic in the university hospital. POPULATION: Fifty-nine patients who were older than 20 years of age and had worked in a full-time job within the past three years. METHODS: All participants underwent clinical evaluation using the visual analog scale (for pain), Shoulder Pain and Disability Index, Pittsburgh Sleep Quality Index, and shoulder ultrasound examination. The work-related ergonomic risks, including dealing with heavy objects, repeated use and requiring forceful motion of the affected upper extremity, were assessed. The ultrasound-identified shoulder pathologies associated with job discontinuation, that is, sick leave due to painful shoulder for more than two consecutive months, were considered as the primary outcome. In the job discontinuation subgroup, we further investigated the association between return to work and the clinical/sonographic findings. RESULTS: Univariate analysis revealed a positive association between job discontinuation and shoulder surgery or work types requiring forceful upper-limb movements. Multivariate analysis demonstrated that job discontinuation was positively associated with supraspinatus tendon full-thickness tears (risk ratio, 8.80; 95% CI, 1.77-10.56; P=0.018). Of the patients who received shoulder surgery, 46.6% had recurrent rotator cuff tears. Return to work was likely to be related to pain scores during overhead activities and shoulder function impairment but not to sonographic findings. CONCLUSIONS: Job discontinuation is associated with shoulder surgery, work that necessitates forceful upper-extremity movements and supraspinatus tendon full-thickness tears detected by ultrasound. CLINICAL REHABILITATION IMPACT: Sonographic findings should not be used as the only standard for evaluating the patient's work capability.

[Microsurgical thinned anterolateral thigh perforator flap to repair soft tissue defects of foot and ankle].

OBJECTIVE: To explore method and clinical effect of microsurgical thinned anterolateral thigh perforator flap to repair soft tissue defects of foot and ankle. METHODS: From March 2017 to January 2022, totally 20 patients with soft tissue defects of ankle joint were treated with micro-thinning anterolateral perforator flap for free transplantation, included 13 males and 7 females, aged from 22 to 58 years old with an average of (36.45±12.36) years old. The size of flap ranged from 8.0 cm×5.0 cm to 20.0 cm×12.0 cm. Before operation, perforating vessels on the anterolateral thigh region were detected and marked with a portable Doppler detector. For the defect width less than 8 cm, 11 patients were repaired with a single flap. For the defect width more than 8 cm, the wound could not be sutured directly, and the lobulated flap technique was used in 9 patients, the width was converted to length, and the donor site was closed directly. Under the microscope, all flaps were thinened in a stepwise manner from the center of the pedicle to the periphery. After operation, survival of the flap, the shape, texture, sensory function recovery were observes, and recovery of foot function was evaluated by Maryland foot function evaluation standard. RESULTS: All 20 patients with microsurgical thinned anterolateral thigh perforator flaps were survived. Venous crisis occurred in 1 patient due to subcutaneous hematoma, after removal of the hematoma, the crisis was relieved and the flap survived successfully. The wounds in the donor and recipient sites healed well, and only linear scars left in the donor sites. Twenty patients were followed up for 3 to 26 months after operation, good shape of flaps without bloated, and good texture. The two-point discrimination of free flaps ranged from 9.0 to 16.0 mm, and the protective sensation was restored. The ankle flexion and extension function recovered well and patients could walk normally. According to Maryland foot function evaluation standard, 8 patients got excellent result, 10 patients good and 2 middle. CONCLUSION: Microsurgical thinned anterolateral thigh perforator flap is an ideal method to repair soft tissue defects in functional area of foot and ankle, with good appearance and texture of the flap, no need for re-plastic surgery, reduced hospitalization costs, and less donor site damage.

Enhancing diagnosis and treatment of superior cluneal nerve entrapment: cadaveric, clinical, and ultrasonographic insights.

OBJECTIVES: Low back pain is a prevalent public health issue caused by superior cluneal nerve (SCN) entrapment. This study aimed to explore the course of SCN branches, cross-sectional area (CSA) of the nerves, and effects of ultrasound-guided SCN hydrodissection. METHODS: SCN distance relative to the posterior superior iliac spines was measured and compared with ultrasound findings in asymptomatic volunteers. The CSA of the SCN, pressure-pain threshold, and pain measurements were obtained from asymptomatic controls and patients with SCN entrapment at various time points after hydrodissection (with 1 mL of 50% dextrose, 4 mL of 1% lidocaine, and 5 mL of 1% normal saline) in the short-axis view. RESULTS: Twenty sides of 10 formalin-fixed cadavers were dissected. The SCN locations on the iliac crest did not differ from the ultrasound findings in 30 asymptomatic volunteers. The average CSA of the SCN across different branches and sites ranged between 4.69-5.67 mm2 and did not vary across different segments/branches or pain statuses. Initial treatment success was observed in 77.7% (n = 28) of 36 patients receiving hydrodissection due to SCN entrapment. A group with initial treatment success experienced symptom recurrence in 25% (n = 7) of cases, and those with recurrent pain had a higher prevalence of scoliosis than those without symptom recurrence. CONCLUSIONS: Ultrasonography effectively localizes SCN branches on the iliac crest, whereby increased nerve CSA is not useful for diagnosis. Most patients benefit from ultrasound-guided dextrose hydrodissection; however, those with scoliosis may experience symptom recurrence and whether structured rehabilitation can reduce recurrence post-injection should be considered as one perspective in future research. Trial registration ClinicalTrials.gov (NCT04478344). Registered on 20 July 2020, https://clinicaltrials.gov/ct2/show/NCT04478344?cond=Superior+Cluneal+Nerve&cntry=TW&draw=2&rank=1 . Critical relevance statement Ultrasound imaging accurately locates SCN branches on the iliac crest, while enlargement of the CSA is not useful in diagnosing SCN entrapment; however, approximately 80% of SCN entrapment cases respond positively to ultrasound-guided dextrose hydrodissection.

Deep learning algorithm for predicting subacromial motion trajectory: Dynamic shoulder ultrasound analysis.

Subacromial motion metrics can be extracted from dynamic shoulder ultrasonography, which is useful for identifying abnormal motion patterns in painful shoulders. However, frame-by-frame manual labeling of anatomical landmarks in ultrasound images is time consuming. The present study aims to investigate the feasibility of a deep learning algorithm for extracting subacromial motion metrics from dynamic ultrasonography. Dynamic ultrasound imaging was retrieved by asking 17 participants to perform cyclic shoulder abduction and adduction along the scapular plane, whereby the trajectory of the humeral greater tubercle (in relation to the lateral acromion) was depicted by the deep learning algorithm. Extraction of the subacromial motion metrics was conducted using a convolutional neural network (CNN) or a self-transfer learning-based (STL)-CNN with or without an autoencoder (AE). The mean absolute error (MAE) compared with the manually-labeled data (ground truth) served as the main outcome variable. Using eight-fold cross-validation, the average MAE was proven to be significantly higher in the group using CNN than in those using STL-CNN or STL-CNN+AE for the relative difference between the greater tubercle and lateral acromion on the horizontal axis. The MAE for the localization of the two aforementioned landmarks on the vertical axis also seemed to be enlarged in those using CNN compared with those using STL-CNN. In the testing dataset, the errors in relation to the ground truth for the minimal vertical acromiohumeral distance were 0.081-0.333 cm using CNN, compared with 0.002-0.007 cm using STL-CNN. We successfully demonstrated the feasibility of a deep learning algorithm for automatic detection of the greater tubercle and lateral acromion during dynamic shoulder ultrasonography. Our framework also demonstrated the capability of capturing the minimal vertical acromiohumeral distance, which is the most important indicator of subacromial motion metrics in daily clinical practice.

Low-Molecular-Weight Chondroitin Sulfates Alleviate Simulated Microgravity-Induced Oxidative Stress and Bone Loss in Mice.

(1) Background: Many studies have shown that microgravity experienced by astronauts or long-term bedridden patients results in increased oxidative stress and bone loss. Low-molecular-weight chondroitin sulfates (LMWCSs) prepared from intact chondroitin sulfate (CS) have been demonstrated to possess good antioxidant and osteogenic activities in vitro. This study aimed to assess the antioxidant activity of the LMWCSs in vivo and evaluate their potential in preventing microgravity-induced bone loss. (2) Methods: we used hind limb suspension (HLS) mice to simulate microgravity in vivo. We investigated the effects of LMWCSs against oxidative stress damage and bone loss in HLS mice and compared the findings with those of CS and a non-treatment group. (3) Results: LMWCSs reduced the HLS-induced oxidative stress level, prevented HLS-induced alterations in bone microstructure and mechanical strength, and reversed changes in bone metabolism indicators in HLS mice. Additionally, LMWCSs downregulated the mRNA expression levels of antioxidant enzyme- and osteogenic-related genes in HLS mice. The results showed that overall effect of LMWCSs was better than that of CS. (4) Conclusions: LMWCSs protect against the bone loss caused by simulated microgravity, which may be related to their ability to reduce oxidative stress. LMWCSs can be envisaged as potential antioxidants and bone loss protective agents in microgravity.

Application of deep learning algorithms in automatic sonographic localization and segmentation of the median nerve: A systematic review and meta-analysis.

OBJECTIVE: High-resolution ultrasound is an emerging tool for diagnosing carpal tunnel syndrome caused by the compression of the median nerve at the wrist. This systematic review and meta-analysis aimed to explore and summarize the performance of deep learning algorithms in the automatic sonographic assessment of the median nerve at the carpal tunnel level. METHODS: PubMed, Medline, Embase, and Web of Science were searched from the earliest records to May 2022 for studies investigating the utility of deep neural networks in the evaluation of the median nerve in carpal tunnel syndrome. The quality of the included studies was evaluated using the Quality Assessment Tool for Diagnostic Accuracy Studies. The outcome variables included precision, recall, accuracy, F-score, and Dice coefficient. RESULTS: In total, seven articles were included, comprising 373 participants. The deep learning and related algorithms comprised U-Net, phase-based probabilistic active contour, MaskTrack, ConvLSTM, DeepNerve, DeepSL, ResNet, Feature Pyramid Network, DeepLab, Mask R-CNN, region proposal network, and ROI Align. The pooled values of precision and recall were 0.917 (95 % confidence interval [CI], 0.873-0.961) and 0.940 (95 % CI, 0.892-0.988), respectively. The pooled accuracy and Dice coefficient were 0.924 (95 % CI, 0.840-1.008) and 0.898 (95 % CI, 0.872-0.923), respectively, whereas the summarized F-score was 0.904 (95 % CI, 0.871-0.937). CONCLUSION: The deep learning algorithm enables automated localization and segmentation of the median nerve at the carpal tunnel level in ultrasound imaging with acceptable accuracy and precision. Future research is expected to validate the performance of deep learning algorithms in detecting and segmenting the median nerve along its entire length as well as across datasets obtained from various ultrasound manufacturers.

ALAS1 associated with goat kidding number trait was regulated by the transcription factor ASCL2 to affect granulosa cell proliferation.

ALAS1 is a member of the α-oxoamine synthase family, which is the first rate-limiting enzyme for heme synthesis and is important for maintaining intracellular heme levels. In the ovary, ALAS1 is associated with the regulation of ovulation-related mitochondrial P450 cytochromes, steroid metabolism, and steroid hormone production. However, there are few studies on the relationship between ALAS1 and reproductive traits in goats. In this study, a mutation located in the promoter region of ALAS1 (g.48791372C>A) was found to be significantly (p < 0.05) associated with the kidding number of Yunshang black goats. Specifically, the mean kidding number in the first three litters and the kidding numbers of all three litters were significantly (p < 0.05) higher in individuals with the CA genotype or AA genotype than in those with the CC genotype. To further investigate the regulatory mechanism of ALAS1, the expression of ALAS1 in goat ovarian tissues with different genotypes was verified by real-time quantitative PCR. The results showed that the expression of ALAS1 was significantly higher in the ovaries of individuals with AA genotype than those with AC and CC genotypes (p < 0.01), and the expression trend of transcription factor ASCL2 was consistent with ALAS1. Additionally, the ALAS1 g.48791372C>A mutation created a new binding site for the transcription factor ASCL2. The luciferase activity assay indicated that the mutation increased the promoter activity of ALAS1. Overexpression of the transcription factor ASCL2 induced increased expression of ALAS1 in goat granulosa cells (p < 0.05). The opposite trend was shown for the inhibition of ASCL2 expression. The results of real-time quantitative PCR, EdU and Cell Counting Kit-8 assays indicated that the transcription factor ASCL2 increased the proliferation of goat granulosa cells by mediating the expression of ALAS1. In conclusion, the transcription factor ASCL2 positively regulated the transcriptional activity and expression levels of ALAS1, altering granulosa cell proliferation and the kidding number in goats.

Gene mutations and chromosomal abnormalities in syndromes with tooth agenesis.

This study aims to review the pathogenic mechanisms and clinical manifestations in syndromes with tooth agenesis (TA). Online Mendelian Inheritance in Man and PubMed databases were searched for a comprehensive review. Previous publications reported complicated aetiologies of syndromic TA. Gene mutations in conserved signalling pathways (WNT, EDA, SHH, FGF, and TGF-ß/BMP) and crucial molecules (PAX9, PIXT2, IRF6, the p53 family, and subunits of RNA polymerase III) are the main causes of syndromic TA. In the process of odontogenesis, antagonistic or synergistic interactions are demonstrated in patients and murine models. Mutations in some genes (WNT10A, WNT10B, AXIN2, ANTXR1, MSX1, EDA, EDAR, and EDARADD) can result in both syndromic and isolated TA. In addition, chromosomal anomalies are also responsible for syndromic TA (Down syndrome, Wolf-Hirschhorn syndrome, Williams syndrome, and Pierre Robin sequence). The causes and manifestations of syndromic TA are highly complex, and this constitutes a clinical challenge. Mutations in signalling pathways and crucial molecules as well as chromosomal anomalies are responsible for syndromic TA. And there are overlaps between the causative genes of syndromic and isolated TA.

Subacromial Motion Metrics in Painful Shoulder Impingement: A Dynamic Quantitative Ultrasonography Analysis.

OBJECTIVES: To explore the subacromial motion metrics in patients with and without subacromial impingement syndrome (SIS) and to investigate whether the abnormality was associated with rotator cuff pathologies. DESIGN: This cross-sectional observational study used dynamic quantitative ultrasonography imaging for shoulder joint assessment. SETTING: Outpatient rehabilitation clinic. PARTICIPANTS: Individuals with SIS on at least 1 shoulder (n=32) and asymptomatic controls (n=32) (N=64). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Frame-by-frame, the humeral greater tuberosity against the lateral edge of the acromion was traced to obtain the minimal vertical acromiohumeral distance (AHD). The rotation angle and radius of the humerus were computed using the least-squares curve fitting method. RESULTS: Approximately two-thirds of the shoulders with SIS did not have any sonographically identifiable rotator cuff pathologies. There was a consistent trend of nonsignificantly increased humeral rotation angles in painful shoulders. The generalized estimating equation demonstrated that the decreased minimal vertical AHD was associated with painful subacromial impingement (ß coefficient: -0.123cm, 95% confidence interval [CI], -0.199 to -0.047). The area under the curve for the minimal vertical AHD to discriminate painful or impinged shoulders ranged from 0.624-0.676. The increased rotation angle (ß coefficient: 10.516°; 95% CI, 3.103-17.929) and decreased rotation radius (ß coefficient: -2.903cm; 95% CI, -5.693 to -0.111) were shown to be significantly related to the presence of supraspinatus tendinopathy. CONCLUSIONS: Shoulders with SIS were characterized by a decreased minimal vertical AHD during dynamic examination. Abnormal subacromial metrics can develop in patients with mild (or no) rotator cuff pathologies. More prospective cohort studies are warranted to investigate the changes in subacromial motion metrics in populations at risk for painful or impinged shoulders.

Exploring training dental implant placement using static or dynamic devices among dental students.

BACKGROUND: Static computer-assisted surgery (s-CAIS) and dynamic computer-assisted implant surgery (d-CAIS) are the main digital approaches in guiding dental implant placement. PURPOSE: The aim of this study was to explore and compare the learning curves for s-CAIS and d-CAIS by beginners. MATERIALS AND METHODS: Three dental students used each dental model for drilling five positions with missing teeth. Operators performed the drilling test for five sets of dental models with an interval of 7 ± 1 days assisted by the d-CAIS system. After a six-month break, the same students performed the drilling test again in the same way but with the s-CAIS system. A total of thirty models were used, and 150 implants were inserted. The operation time and relative deviations were recorded and calculated. Correlations between various deviation parameters and attempts were tested with independent-samples Kruskal-Wallis tests. RESULTS: A significant difference between the two groups was found in the operation time (p < .001). For accuracy, the difference was found in the first attempt of coronal and apical deviations but disappeared as the training went on. As the practice progressed, improvement was evident in the d-CAIS group but not in the s-CAIS group. When reaching the plateau stage of the learning curve of the d-CAIS group (after five attempts), the influence of different methods of guidance was limited between the two groups. CONCLUSIONS: A learning curve effect was found in d-CAIS but not in s-CAIS in vitro tests by beginners. The operating procedure of dynamic navigated and static template-guided implant placement was easy to master.

Identification of WNT4 alternative splicing patterns and effects on proliferation of granulosa cells in goat.

Increasing ovulation numbers is one of the most important ways to promote reproduction in mammals, and follicular granulosa cells (GCs) provide the necessary nutrients and microenvironment for oocytes to ovulate. WNT4 has been shown to be a key factor in regulating the proliferation of GCs in mammalian ovarian tissues. Our previous transcriptome sequencing (RNA-seq) results have identified two alternatively spliced products of WNT4;however, little is known about the splicing mechanism and its effect on GC proliferation. In this study, two alternatively spliced products of WNT4, designated WNT4-α and WNT4-ß, were identified by cloning and analyzed for their function by bioinformatics. The RT-qPCR and Western blot results showed that the expression of WNT4-α was significantly higher than that of WNT4-ß in the ovary tissues and GCs of Yunshang black goats. We therefore hypothesized that WNT4-α was the main isoform affecting the proliferation of goat GCs. Subsequently, goat GC proliferation assays showed that overexpression of WNT4-α significantly promoted GC proliferation, and the opposite was true after WNT4-α inhibition. The expression of marker genes of the Wnt signaling pathway was also examined and WNT4-α was found to affect the proliferation and hormone secretion of goat GCs by regulating the Wnt signaling pathway. In addition, a series of splicing factors were involved in in the alternative splicing; in this study, SRSF6 was found to be involved as a splicing factor in the generation of WNT4 alternative splicing. In summary, WNT4 alternative splicing was mediated by the splicing factor SRSF6, and WNT4-α alternative splicing played an important role in follicle development and had a significant effect on the proliferation of goat GCs. The results of this study provide a theoretical foundation for further understanding the molecular regulatory mechanisms of the WNT4 in follicle development in goats.

Analysis of the Association of Two SNPs in the Promoter Regions of the PPP2R5C and SLC39A5 Genes with Litter Size in Yunshang Black Goats.

Screening for candidate genes and genetic variants associated with litter size is important for goat breeding. The aim of this study was to analyze the relationship between single nucleotide polymorphisms (SNPs) in PPP2R5C and SLC39A5 and litter size in Yunshang black goats. KASP genotyping was used to detect the SNP genetic markers in the PPP2R5C and SLC39A5 in a population of 569 Yunshang black goats. The results show that there were two SNPs in the PPP2R5C and SLC39A5 promoter regions. Association analysis revealed that the polymorphisms PPP2R5C g.65977743C>T and SLC39A5 g.50676693T>C were significantly associated with the litter size of the third parity of Yunshang black goats (p < 0.05). To further explore the regulatory mechanism of the two genes, the expression of different genotypes of PPP2R5C and SLC39A5 was validated by RT-qPCR and Western blotting. The expression of PPP2R5C was significantly higher in individuals with the TT genotype than in those with the TC and CC genotypes (p < 0.05). The expression of SLC39A5 was also significantly higher in individuals with the TT genotype than in TC and CC genotypes (p < 0.05). Dual luciferase reporter analysis showed that the luciferase activity of PPP2R5C-C variant was significantly higher than that of PPP2R5C-T variant (p < 0.05). The luciferase activity of SLC39A5-T variant was significantly higher than that of SLC39A5-C variant (p < 0.05). Software was used to predict the binding of transcription factors to the polymorphic sites, and the results show that SOX18, ZNF418, and ZNF667 and NKX2-4 and TBX6 might bind to PPP2R5C g.65977743C>T and SLC39A5 g.50676693T>C, respectively. These results provide new insights into the identification of candidate genes for marker-assisted selection (MAS) in goats.

Expression and potential role of FOSB in glioma.

Background: FOSB is reported to be an oncogene in a variety of tumors. However, the expression and role of FOSB in glioma remain obscure. In this study, we aimed to explore the expression of FOSB in glioma and its biological role in glioblastoma multiforme (GBM). Methods: Western blot, immunohistochemical staining, and quantitative real-time polymerase chain reaction (RT-qPCR) were used to detect the expression of FOSB in clinical samples. FOSB was knocked down in cells to determine the effects of FOSB on the phenotypic changes of tumors by plate cloning, CCK-8 assay, and Transwell assay. Finally, subcutaneous tumorigenesis in nude mice was used to observe the tumorigenesis of glioma cell lines after the knockdown of the FOSB gene. Results: FOSB expression was higher in glioma compared with normal brain tissue. After the downregulation of FOSB, the expression of cleaved caspase-3 increased. Plate cloning and CCK-8 experiments showed that the proliferation of glioma cell lines decreased. The Transwell assay demonstrated that the glioblastoma cell lines had lower migration ability after the knockdown of FOSB. Finally, the tumor volume of U87 glioma cells in group sh-FOSB was smaller than that in the control group. The TUNEL staining in vitro showed that the apoptosis of sh-FOSB glioma cells increased. Conclusion: FOSB was highly expressed in glioma tissues. The viability of glioma cells decreased, and the ability of glioma cells to proliferate and migrate was reduced when FOSB was downregulated. Hence, FOSB may promote the development and migration of gliomas.

Effect of Upregulation of Transcription Factor TFDP1 Binding Promoter Activity Due to RBP4 g.36491960G>C Mutation on the Proliferation of Goat Granulosa Cells

Retinol-binding protein 4 (RBP4), a member of the lipocalin family, is a specific carrier of retinol (vitamin A) in the blood. Numerous studies have shown that RBP4 plays an important role in mammalian embryonic development and that mutations in RBP4 can be used for the marker-assisted selection of animal reproductive traits. However, there are few studies on the regulation of reproduction and high-prolificacy traits by RBP4 in goats. In this study, the 5' flanking sequence of RBP4 was amplified, and a G>C polymorphism in the promoter region -211 bp (g.36491960) was detected. An association analysis revealed that the respective first, second and third kidding number and mean kidding number of nanny goats with CC and GC genotypes (2.167 ± 0.085, 2.341 ± 0.104, 2.529 ± 0.107 and 2.189 ± 0.070 for CC and 2.052 ± 0.047, 2.206 ± 0.057, 2.341 ± 0.056 and 2.160 ± 0.039 for GC) were significantly higher (p < 0.05) than those with the GG genotype (1.893 ± 0.051, 2.027 ± 0.064, 2.107 ± 0.061 and 1.74 ± 0.05). The luciferase assay showed that luciferase activity was increased in C allele individuals compared with that in G allele individuals. A competitive electrophoretic mobility shift assay (EMSA) showed that individuals with the CC genotype had a stronger promoter region binding capacity than those with the GG genotype. In addition, transcription factor prediction software showed that the RBP4 g.36491960G>C mutation added a novel binding site for transcription factor DP-1 (TFDP1). RT−qPCR results showed that the expression of TFDP1 was significantly higher in the high-prolificacy group than in the low-prolificacy group, and the expression of RBP4 was higher in both the CC and GC genotypes than that in the GG genotype. TFDP1 overexpression significantly increased the expression of RBP4 mRNA (p < 0.05) and the expression of the cell proliferation factors cyclin-D1, cyclin-D2 and CDK4 (p < 0.05). The opposite trend was observed after interference with TFDP1. Both the EdU and CCK-8 results showed that TFDP1 expression could regulate the proliferation of goat ovarian granulosa cells. In summary, our results showed that RBP4 g.36491960G>C was significantly associated with fecundity traits in goats. The g.36491960G>C mutation enhanced the transcriptional activity of RBP4 and increased the expression of RBP4, thus improving the fertility of Yunshang black goats.

Ultrasound Imaging of Facial Vascular Neural Structures and Relevance to Aesthetic Injections: A Pictorial Essay.

The facial and submental regions are supplied by complicated neurovascular networks; therefore, facial aesthetic injections may be associated with serious adverse events such as skin necrosis and blindness. Pre-injection localization of neurovascular structures using high-resolution ultrasound can theoretically prevent unexpected complications. Therefore, a systematic protocol that focuses on these facial neurovascular structures is warranted. In this pictorial essay, we discuss the sonoanatomy of facial and submental neurovascular structures and its relevance to aesthetic injections. Moreover, we have highlighted the mechanisms underlying potential neurovascular injuries during aesthetic injections.

Transcriptome Analysis Reveals Key miRNA-mRNA Pathways in Ovarian Tissues of Yunshang Black Goats With Different Kidding Numbers.

The granulosa cell (GC) is the basic functional unit of follicles, and it is important for promoting follicle growth and sex hormones, as well as growth factor secretion in the process of reproduction. A variety of factors influence granulocyte proliferation, yet there are still many gaps to be filled in target and non-coding RNA regulation. In our study, the differentially expressed (DE) mRNAs and miRNAs were detected by using RNA-seq, and we constructed a mRNA-miRNA network related to goat prolificacy. Then, the goat primary GCs were isolated from the follicle for the function validation of candidate genes and their regulator miRNAs. A total of 2,968 DE mRNAs and 99 DE miRNAs were identified in the high- and low-prolificacy goat by RNA-seq, of which there were 1,553 upregulated and 1,415 downregulated mRNAs, and 80 upregulated and 19 downregulated miRNAs, respectively. JAK3 was identified as highly expressed in the low-prolificacy goats (3 times higher than high-prolificacy goats), and the integrated analysis showed that chi-miR-493-3p was a potential regulator of JAK3. The analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) showed that JAK3 was involved in the PI3K-Akt signaling pathway, the Jak-STAT signaling pathway, and signaling pathways regulating pluripotency of stem cells. In particular, the PI3K-Akt signaling pathway was a typical pathway for cell proliferation, differentiation, apoptosis, and migration. We found that the chi-miR-493-3p targets JAK3 directly via RT-qPCR, dual fluorescence assays, and Western blot. Furthermore, the expression of JAK3 was significantly decreased by the chi-miR-493-3p mimic and increased by the chi-miR-493-3p inhibitor. The CCK-8 assay showed that overexpression of JAK3 promoted cell proliferation, while inhibiting JAK3 had the opposite effect. The expression of cell proliferation markers CDK4 and cyclin D2 also showed the same results. Moreover, the enzyme-linked immunosorbent assay showed that steroid hormones E2 and PROG were increased by overexpressing JAK3 and decreased by inhibiting JAK3. Therefore, our results identified a chi-miR-439-3p-JAK3 regulatory pathway, which provided a new insight into the GC proliferation and prolificacy of goat.

A Novel SNP in the Promoter Region of IGF1 Associated With Yunshang Black Goat Kidding Number via Promoting Transcription Activity by SP1.

IGF1, a member of the insulin-like growth factor (IGF) superfamily, is also known as the growth-promoting factor (somatomedin C). IGF1 is involved in vertebrate growth and development, immunity, cell metabolism, reproduction, and breeding. However, there are relatively few studies on the relationship between IGF1 and goat reproduction. In this study, a new transcription factor SP1 bound to the IGF1 g. 64943050T>C promoted granulosa cell (GC) proliferation. A mutation g.64943050T>C located in the promoter region of IGF1 was identified. Association analysis revealed that the kidding number in the first and second litters and the average number of first three litters of the CC genotype (2.206 ± 0.044, 2.254 ± 0.056, and 2.251 ± 0.031) were significantly higher than those in the TC genotype (1.832 ± 0.049, 1.982 ± 0.06, and 1.921 ± 0.034) and TT genotype (1.860 ± 0.090, 1.968 ± 0.117, and 1.924 ± 0.062) (p < 0.05). The kidding number in the third litter of the CC genotype (2.355 ± 0.057) was significantly higher than that in the TT genotype (2.000 ± 0.107) (p < 0.05). Then, the function of this mutation was validated by the dual-luciferase reporter assay and EMSA. The results showed that the luciferase activity of IGF1-mutant-C was significantly higher than that of IGF1-Wild-T (p < 0.05). The EMSA also showed that the binding ability of IGF1-mutant-C was higher than that of IGF1-Wild-T (p < 0.05). Subsequently, the transcription factor SP1 was predicted to bind to the mutation of IGF1 (g.64943050T>C). Overexpression of SP1 promotes the expression of IGF1 in the primary granulosa cells (GCs). The results of the CCK-8 assay and the expression of GC proliferation factors (CDK4, cyclin D1, and cyclin D2) demonstrated that SP1 promoted GC proliferation by regulating IGF1 expression. Our results suggested that the IGF1 g.64943050T>C was significantly associated with the kidding number of Yunshang black goats, and SP1 as a transcription factor of IGF1 binding to the mutation T>C regulated the expression of IGF1. Furthermore, SP1 promoted goat GC proliferation by regulating the expression of IGF1, which provides a new insight for the goat fertility trait.