[Euonymus alatus delays progression of diabetic kidney disease in mice by regulating EGFR tyrosine kinase inhibitor resistance signaling pathway].

OBJECTIVE: To explore the therapeutic mechanism of Euonymus alatus for diabetic kidney disease (DKD). METHODS: TCMSP, PubChem and Swiss Target Prediction databases were used to obtain the active ingredients in Euonymus alatus and their targets. GEO database and R language were used to analyze the differentially expressed genes in DKD. The therapeutic targets of DKD were obtained using GeneCards, DisGeNet, OMIM and TTD databases. The protein-protein interaction network and the "drug-component-target-disease" network were constructed for analyzing the topological properties of the core targets, which were functionally annotated using GO and KEGG pathway enrichment analyses. Molecular docking was performed for the core targets and the main pharmacologically active components, and the results were verified in db/db mice. RESULTS: Analysis of GSE96804, GSE30528 and GSE30529 datasets (including 60 DKD patients and 45 normal samples) identified 111 differentially expressed genes in DKD. Network pharmacology analysis obtained 161 intersecting genes between the target genes of Euonymus alatus and DKD, including the key core target genes SRC, EGFR, and AKT1. The core active ingredients of Euonymus alatus were quercetin, kaempferol, diosmetin, and naringenin, which were associated with responses to xenobiotic stimulionus and protein phosphorylation and regulated EGFR tyrosine kinase inhibitor resistance pathways. Molecular docking suggested good binding activities of the core active components of Euonymus alatus with the core targets. In db/db mouse models of DKD, treatment with Euonymus alatus obviously ameliorated kidney pathologies, significantly inhibited renal expressions of SRC, EGFR and AKT1, and delayed the progression of DKD. CONCLUSION: Euonymus alatus contains multiple active ingredients such as quercetin, kakaferol, diosmetin, naringenin, which regulate the expressions of SRC, EGFR, and AKT1 to affect the EGFR tyrosine kinase inhibitor resistance signaling pathway to delay the progression of DKD.

Clinical and molecular characterization of patients with YWHAG-related epilepsy.

OBJECTIVE: YWHAG variant alleles have been associated with a rare disease trait whose clinical synopsis includes an early onset epileptic encephalopathy with predominantly myoclonic seizures, developmental delay/intellectual disability, and facial dysmorphisms. Through description of a large cohort, which doubles the number of reported patients, we further delineate the spectrum of YWHAG-related epilepsy. METHODS: We included in this study 24 patients, 21 new and three previously described, with pathogenic/likely pathogenic variants in YWHAG. We extended the analysis of clinical, electroencephalographic, brain magnetic resonance imaging, and molecular genetic information to 24 previously published patients. RESULTS: The phenotypic spectrum of YWHAG-related disorders ranges from mild developmental delay to developmental and epileptic encephalopathy (DEE). Epilepsy onset is in the first 2 years of life. Seizure freedom can be achieved in half of the patients (13/24, 54%). Intellectual disability (23/24, 96%), behavioral disorders (18/24, 75%), neurological signs (13/24, 54%), and dysmorphisms (6/24, 25%) are common. A genotype-phenotype correlation emerged, as DEE is more represented in patients with missense variants located in the ligand-binding domain than in those with truncating or missense variants in other domains (90% vs. 19%, p < .001). SIGNIFICANCE: This study suggests that pathogenic YWHAG variants cause a wide range of clinical presentations with variable severity, ranging from mild developmental delay to DEE. In this allelic series, a genotype-phenotype correlation begins to emerge, potentially providing prognostic information for clinical management and genetic counseling.

Combining the triglyceride-glucose index and glycated hemoglobin A1c to assess the risk of preeclampsia in women with normal glucose tolerance: a cross-sectional study.

OBJECTIVE: This study aimed to explore the relationship between the triglyceride-glucose (TyG) index, glycated hemoglobin A1c (HbA1c), and preeclampsia in pregnant women without gestational diabetes mellitus (GDM). PATIENTS AND METHODS: This retrospective study included pregnancies with normal oral glucose tolerance tests (OGTTs) from March 2018 to February 2019. During the second trimester, serum lipids, fasting plasma glucose (FPG), and HbA1c were measured, and OGTTs were performed. Participants were classified into four groups based on their TyG index and HbA1c levels. Logistic regression analysis was done to determine the odds ratios (ORs), and receiver operating characteristic (ROC) curve analysis was used to evaluate the ability of the TyG index and HbA1c to predict the risks of preeclampsia. RESULTS: Patients with preeclampsia exhibited higher TyG index and HbA1c levels (all p < 0.001). The incidence of preeclampsia increased with elevated TyG index and HbA1c levels individually. Furthermore, the highest incidence of preeclampsia was observed when both the TyG index and HbA1c levels were elevated. ROC curve analysis revealed that the combined TyG index and HbA1c displayed an area under the curve (AUC) of 0.689 in predicting the risk of preeclampsia. Even after adjusting for potential confounding factors, the risk of developing preeclampsia remained significantly higher. These associations were especially prominent in women aged ≥ 35 years or those with a normal BMI. CONCLUSIONS: The findings of this study indicate that increased TyG index and HbA1c levels are associated with a higher incidence and risk of preeclampsia in women with normal glucose tolerance during pregnancy. The TyG index and HbA1c levels may serve as potential markers for preeclampsia in individuals with normal OGTT results.

A Variant in the IRF6 Promoter Associated with the Risk for Orofacial Clefting.

The single-nucleotide polymorphism (SNP) rs2235371 (IRF6 V274I) is associated with nonsyndromic cleft lip with or without cleft palate (NSCL/P) in Han Chinese and other populations but appears to be without a functional effect. To find the common etiologic variant or variants within the haplotype tagged by rs2235371, we carried out targeted sequencing of an interval containing IRF6 in 159 Han Chinese with NSCL/P. This study revealed that the SNP rs12403599, within the IRF6 promoter, is associated with all phenotypes of NSCL/P, especially nonsyndromic cleft lip (NSCLO) and a subphenotype of it, microform cleft lip (MCL). This association was replicated in 2 additional much larger cohorts of cases and controls from the Han Chinese. Conditional logistic analysis indicated that association of rs2235371 with NSCL/P was lost if rs12403599 was excluded. rs12403599 contributes the most risk to MCL: its G allele is responsible for 38.47% of the genetic contribution to MCL, and the odds ratios of G/C and G/G genotypes were 2.91 and 6.58, respectively, for MCL. To test if rs12403599 is functional, we carried out reporter assays in a fetal oral epithelium cells (GMSM-K). Unexpectedly, the risk allele G yielded higher promoter activity in GMSM-K. Consistent with the reporter studies, expression of IRF6 in lip tissues from NSCLO and MCL patients with the G/G phenotype was higher than in those from patients with the C/C phenotype. These results indicate that rs12403599 is tagging the risk haplotype for NSCL/P better than rs2235371 in Han Chinese and supports investigation of the mechanisms by which the allele of rs12403599 affects IRF6 expression and tests of this association in different populations.

Reference intervals of systemic immune-inflammation index, neutrophil-to-lymphocyte ratio, lymphocyte-to-monocyte ratio, and platelet-to-lymphocyte ratio during normal pregnancy in China.

OBJECTIVE: To observe the changes in systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR) during normal pregnancy and establish appropriate reference intervals (RIs) for healthy pregnant women. PATIENTS AND METHODS: This retrospective study was conducted from March 2018 to February 2019. Blood samples were collected from healthy pregnant and nonpregnant women. The complete blood count (CBC) parameters were measured, and SII, NLR, LMR, and PLR were calculated. RIs were established using the 2.5th and 97.5th percentile of the distribution. Besides, the differences in CBC parameters between three pregnant trimesters and maternal ages were also compared to assess their influences on each indicator. RESULTS: SII and NLR in three pregnant trimesters increased in pregnant women, and the upper limit of SII and NLR in trimester 2 showed the highest value. On the contrary, LMR decreased in all three pregnant trimesters compared with nonpregnant women, and the values of LMR and PLR showed a gradual downward trend along with the trimesters. Besides, RIs of SII, NLR, LMR, and PLR during different trimesters in different age partitions showed that the values of SII, NLR, and PLR increased with age in a general trend, while LMR showed the opposite trend (p < 0.05). CONCLUSIONS: The SII, NLR, LMR, and PLR showed dynamic changes during pregnant trimesters. RIs of SII, NLR, LMR, and PLR for healthy pregnant women according to pregnant trimesters and maternal age were established and validated in this study, which will promote the standardization of clinical application.

[Effect of laparoscopic ovarian cystectomy on ovarian reserve and the outcomes of in vitro fertilization patients with benign ovarian cysts].

Objective: To investigate the effect of laparoscopic ovarian cystectomy on anti-Mullerian hormone (AMH) level, ovarian response to gonadotropin stimulation and pregnancy rate for in vitro fertilization (IVF) patients with benign ovarian cysts. Methods: Patients with benign ovarian cysts who were admitted for cystectomy and had undergone IVF treatment were enrolled in the study. There were 373 participants with ovarian cysts underwent laparoscopic ovarian cystectomy in the experimental group. According to duration of post-surgery, there were four sub-groups: 1 year post-surgery (1Y POST), 2 years post-surgery (2Y POST), 4 years post-surgery (4Y POST) and ≥5 years post-surgery (≥5Y POST) in the experimental group. According to histopathologic types of ovarian cysts, there were two sub-groups: ovarian endometriotic cysts and ovarian non-endometriotic cysts. Two hundreds and three patients with no history of ovarian cysts and ovarian surgery were in the control group. The level of AMH and basic concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P), estradiol (E2) were measured. Antral follicle counts (AFC) were calculated. There were other study variables: total dose of gonadotropins, duration of ovarian stimulation, the number of oocyte retrieved, the number of embryo obtained, blastocyst transfer rate and pregnancy rate. Results: The control group was matched as closely as possible to the experimental group, including age, body mass index and menstrual cycle (all P>0.05). Compared to the women in control group, the women in ovarian endometriotic cystectomy sub-group had significantly higher levels of basal FSH and basal P, lower level of AMH (all P<0.05); the women in ovarian endometriotic cysts sub-group had significantly higher dose of gonadotropins (all P<0.05); the women in ovarian endometriotic cysts ≥5Y POST sub-group had significantly lower number of oocyte retrieved, lower number of embryo obtained, lower blastocyst transfer rate, and lower pregnancy rate (all P<0.05). Compared to the women in control group, the women in ovarian non-endometriotic cysts sub-group had a significantly higher level of basal FSH and basal P (all P<0.05). The women in ovarian non-endometriotic cysts sub-group had lower level of AMH, higher dose of gonadotropins, lower number of oocyte retrieved, lower number of embryo obtained, lower rate of blastocyst transfer and lower rate of pregnancy than the control group but there were no statistically significant differences among them (all P>0.05). The women with unilateral ovarian endometriotic cysts had significantly lower number of oocyte retrieved on the side of surgery than another side (P<0.05). Conclusions: In short term laparoscopic ovarian cystectomy has no significant effect on ovarian reserve. But with long-term follow-up ovarian reserve, ovarian response to gonadotropin stimulation and pregnancy rate are decreased. The effect of laparoscopic ovarian cystectomy in benign cysts on ovarian is associated with whether or not it is the surgical side.

[Correlation between serum IgM antibody and viral load with clinical symptoms in neonates infected with cytomegalovirus].

To investigate the correlation between serum cytomegalovirus (CMV) IgM antibody/viral load and infection-related clinical symptoms in neonates infected with CMV, and provide basis for clinical assessment and monitoring of neonatal CMV infection. A total of 70 neonates with CMV infection admitted to neonatology in Women's Hospital, School of Medicine Zhejiang University, from January 2014 to December 2020 were included in this study. Using real-time quantitative PCR as the diagnostic criteria, congenital cytomegalovirus-infected neonates (n=29) was diagnosed within the first 3 weeks of life, otherwise, it was postnatally acquired cytomegalovirus infection (n=41). The differences in general information and clinical indicators between IgM antibody positive and negative patients were analyzed, combined with the PCR result, the correlation between the IgM/viral load and the occurrence of symptoms were analyzed. T-test and non-parametric test were used to compare the differences of indicators between groups, logistic regression was used for multivariate analysis, and ROC curve was used to evaluate the auxiliary diagnostic value of relevant indicators. In the congenital CMV infection group and the postnatally acquired CMV infection group, viral load and the proportion of symptomatic patients in IgM positive group were significantly higher than IgM negative group (Z=-2.616, P=0.008; 80% vs. 21%, P=0.005) (Z=-2.405, P=0.016; 56% vs. 19%, P=0.025). Logistic regression analysis of the included population showed the risk factors of CMV infection-related symptoms were IgM positive (OR 4.562, 95%CI:1.461-14.246,P=0.009) and viral load (OR 1.728, 95%CI:1.068-2.798,P=0.026). Regressive analysis for single symptom with correction showed IgM antibody positive was associated with hearing dysfunction(OR 3.954, 95%CI:1.066-14.677,P=0.040),the CMV viral load was associated with thrombocytopenia (OR 2.228, 95%CI:1.124-4.413,P=0.022), and brain imaging abnormalities (OR 3.956, 95%CI:1.421-11.011, P=0.008). Receiver operating characteristic (ROC) analysis showed the area under ROC curve of CMV viral load for brain imaging abnormalities was 0.883 (P<0.001), with a sensitivity of 75.0% and specificity of 90.3%. For neonates infected with CMV, the risk of infection-related clinical symptoms and hearing dysfunction may be increased when IgM antibody was positive. Meanwhile, the higher the CMV viral load at diagnosis, the higher the risk of thrombocytopenia and abnormal brain imaging.

Differentiation of transverse sinus thrombosis from congenitally atretic transverse sinus with time-resolved contrast-enhanced magnetic resonance angiography.

AIM: To establish the value of time-resolved contrast-enhanced magnetic resonance angiography (TR-MRA) in identifying transverse sinus (TS) thrombosis in patients with a unilaterally absent TS signal on phase-contrast (PC) magnetic resonance venography (MRV). MATERIALS AND METHODS: Forty-five patients who were suspected of TS thrombosis clinically with a unilaterally absent TS signal on PC MRV were evaluated using TR-MRA. The image quality was compared between PC MRV and TR-MRA via calculating signal-to-noise ratios (SNRs) and contrast-to-noise ratios (CNRs) of the normal TS. Images obtained by PC MRV and TR-MRA were assessed independently by two neuroradiologists regarding whether the absent signal was thrombosis or hypoplastic TS. Then a consultant neurologist made the final diagnosis based on all available clinical, laboratory, and radiological results (delivered by a consultant neuroradiologist via computed tomography [CT]/CT venography, magnetic resonance imaging [MRI]/MRV or digital subtraction angiography [DSA]), which was set as the reference standard. The accuracy of the radiological diagnosis from both techniques was assessed. RESULTS: For image quality, the SNRs and CNRs of TR-MRA were 452.14 and 440.92, respectively, significantly higher than that of PC MRV (both p<0.001). The interobserver agreement of TR-MRA in identifying TS thrombosis from hypoplastic TS was excellent (κ = 0.951; 95% confidence interval [CI], 0.902-1), much higher than PC MRV (κ = 0.526; 95% CI, 0.389-0.663). Consensus of assessment based on TR-MRA was highly consistent with the reference standard (100% sensitivity, 96.67% specificity) and superior to PC MRV (60% sensitivity, 90% specificity). CONCLUSION: TR-MRA is better than PC MRV for visualising TS and distinguishing thrombosis from congenitally atretic TS.

Aetiology of tinea capitis in China: a multicentre prospective study.

BACKGROUND: Tinea capitis is still common in developing countries, such as China. Its pathogen spectrum varies across regions and changes over time. OBJECTIVES: This study aimed to clarify the current epidemiological characteristics and pathogen spectrum of tinea capitis in China. METHODS: A multicentre, prospective descriptive study involving 29 tertiary hospitals in China was conducted. From August 2019 to July 2020, 611 patients with tinea capitis were enrolled. Data concerning demography, risk factors and fungal tests were collected. When necessary, the pathogens were further identified by morphology or molecular sequencing in the central laboratory. RESULTS: Among all enrolled patients, 74·1% of the cases were in patients aged 2-8 years. The children with tinea capitis were mainly boys (56·2%) and were more likely than adults to have a history of animal contact (57·4% vs. 35·3%, P = 0·012) and zoophilic dermatophyte infection (73·5% vs. 47%). The adults were mainly female (83%) and were more likely than children to have anthropophilic agent infection (53% vs. 23·9%). The most common pathogen was zoophilic Microsporum canis (354, 65·2%), followed by anthropophilic Trichophyton violaceum (74, 13·6%). In contrast to the eastern, western and northeastern regions, where zoophilic M. canis predominated, anthropophilic T. violaceum predominated in central China (69%, P < 0·001), where the patients had the most tinea at other sites (20%) and dermatophytosis contact (26%) but the least animal contact (39%). Microsporum ferrugineum was the most common anthropophilic agent in the western area, especially in Xinjiang province. CONCLUSIONS: Boys aged approximately 5 years were the most commonly affected group. Dermatologists are advised to pay more attention to the different transmission routes and pathogen spectra in different age groups from different regions.

[Analysis of chronic obstructive pulmonary disease screening questionnaire and pulmonary function test among migrant workers exposed to dust].

Objective: To analyze the application of Chronic Obstructive Pulmonary Disease (COPD) Screening Questionnaire and pulmonary function test in dust-exposed migrant workers. Methods: In May 2019, 149 cases of dust exposed migrant workers were selected as the research subjects through the free clinic in the countryside. COPD Screening Questionnaire and lung function test were carried out to analyze the high-risk groups and the influencing factors of positive pulmonary function test results. Results: Among 149 cases of dust-exposed migrant workers, 107 (71.8%) were positive for questionnaire screening, 73 (49.0%) were positive for pulmonary function test, 75 (50.3%) were diagnosed with coal worker's pneumoconiosis, and 101 (67.8%) were diagnosed with lung function injury. The positive rate of pulmonary function of migrant workers with positive questionnaire screening results was significantly higher than that of those with negative results (P<0.05) . The results of multivariate analysis showed that compared with non-pneumoconiosis, the risk of positive pulmonary function test results was higher in dust-exposed migrant workers with stage Ⅲ pneumoconiosis (OR=16.462, 95%CI: 3.390-79.946; P<0.01) . Compared with non-smoking, the risks of positive pulmonary function test results of dust-exposed migrant workers with smoking index of 11-20 package years and >20 package years were higher (OR=19.814, 95%CI: 3.854-101.883; OR=9.733, 95%CI: 2.310-41.008; P<0.01) . Conclusion: The risk of COPD in dust-exposed migrant workers is high, so we should strengthen the early examination of the high pneumoconiosis stage and smoking population. The screening questionnaire can better screen out the high-risk groups of COPD, and it can be used as a basic screening tool.

Integrated microRNA-mRNA analysis reveals the roles of microRNAs in the muscle fat metabolism of Yanbian cattle.

Fat deposition is an important economic trait in farm animals. However, it is difficult to genetically improve intramuscular fat deposition via trait-based cattle breeding. The main objectives of this study were to analyze the factors about beef flavor, and to detect functional microRNA (miRNA, miR) associated with intramuscular fat deposition in Yanbian cattle. Longissimus dorsi samples from six steers were separated into high- and low-fat groups (n = 3 each) based on the marbling score, and transcriptomic analysis was performed using miRNA sequencing. A total of 33 miRNAs and 38 genes were found to be differentially expressed in the high- and low-fat groups. Quantitative real-time polymerase chain reaction was performed to validate the sequencing results. Integrated miRNA-mRNA analysis revealed that miRNA-associated target genes were primarily associated with skeletal muscle development. However, some of the miRNAs (miR-424 etc.) and genes (ATF3 etc.) were also associated with fat metabolism. A targeted relationship between miR-22-3p and the WFIKKN2 gene and its involvement in adipocyte differentiation were confirmed experimentally. The study findings may provide potential candidate molecular targets for the selection of cattle with improved meat quality.

[Comparative analysis of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis in paraffin-embedded tissue of clinical tuberculous wound].

Objective: To observe the effect of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis (MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods: The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang'an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher's exact probability test, one-way analysis of variance, independent sample t test and Wilcoxon signed rank test. Results: The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups (P=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar (F=1.284, P=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining (t=-12.495, -7.961, P<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue (Z=-3.162, P<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining (t=4.569, 15.519, P<0.01). Conclusions: Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.

Bovine bta-microRNA-1271 Promotes Preadipocyte Differentiation by Targeting Activation Transcription Factor 3.

Yanbian yellow cattle are one of the top five largest breeds of cattle in China. We had previously found that bta-miR-1271 is differentially expressed in the longissimus dorsi muscles of Yanbian yellow bulls and steers. However, whether bta-miR-1271 affects bovine fat formation is unclear. In this study, we used target gene prediction, dual-luciferase reporter assay, and transfection-mediated overexpression and inhibition of bta-miR-1271 in a culture of Yanbian yellow cattle preadipocytes to investigate the role of bta-miR-1271 in adipogenesis. We showed that bta-miR-1271 directly targets the 3'-untranslated region (3'-UTR) of the activating transcription factor 3 (ATF3) mRNA and downregulates its expression. Overexpression of bta-miR-1271 enforced by the miRNA mimics promoted triglyceride accumulation and significantly upregulated expression of the adipogenic peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT enhancer-binding protein α (C/EBPα) genes at both the protein and mRNA levels, as demonstrated by RT-qPCR and Western blot analyses. Conversely, inhibition of bta-miR-1271 expression produced the opposite effect. Our results show that bta-miR-1271 regulates differentiation of Yanbian yellow cattle preadipocytes by inhibiting ATF3 expression, which highlights the importance of microRNA-mediated regulation of adipogenesis. miR-1271 and its target gene(s) may provide a new research direction for investigating biological agents affecting intramuscular fat deposition in cattle.

[Research advances in healing mechanism of chronic refractory wounds mediated by long non-coding RNA].

Chronic refractory wound refers to the wound with unclear etiology, multiple and complex injury factors, slow healing, and no obvious tendency of healing after treatment for 4 weeks. The formation and evolution process of chronic refractory wounds are very complex, involving re-epithelialization of wound tissue, cell proliferation, tissue remodeling, and angiogenesis and lymphangiogenesis. The abnormal expression of long non-coding RNA may be involved in the formation of chronic refractory wounds, but the specific pathogenesis and related molecular biological changes are still controversial. In this paper, we reviewed the process and role of long non-coding RNA in regulating keratinocyte differentiation, fibroblast proliferation, and regeneration of vascular and lymphatic endothelial cell in chronic refractory wounds.

CircRNA_009934 induces osteoclast bone resorption via silencing miR-5107.

OBJECTIVE: We aimed to explore the expression of circRNA_009934 in osteoclast, as well as its potential roles in regulating osteoclastogenesis and bone resorption via regulating miR-5107. MATERIALS AND METHODS: We performed qRT-PCR analysis to examine the expression of circRNA_009934 in osteoclast in distinctive stages. We used CCK-8 assay to detect the cell proliferation ability. Correlation analysis between the expression levels of circRNA_009934 and miR-5107 was performed using statistical analysis. Bioinformatics prediction was performed to predict the binding site of circRNA_009934 and miR-5107, subsequently followed by Luciferase assay for validation. The mice TRAF6 3'-UTR were cloned into the Luciferase reporter vector and miR-5107 binding mutants were constructed to validate the inhibited regulation of miR-5107 to the expression of TRAF6. RESULTS: Our results showed that expression of circRNA_009934 was increased during osteoclast differentiation. CircRNA_009934 expression was closely correlated with osteoclastogenesis and bone resorption activity. Bioinformatics prediction and Luciferase assay demonstrated that circRNA_009934 served as a ceRNA of miR-5107 and regulated its downstream TRAF6 expression. CONCLUSIONS: We first demonstrated that circRNA_009934 expression was increased in osteoclasts, which promoted osteoclastogenesis by serving as a ceRNA of miR-5107 and regulated the expression of TRAF6.

Traumatic fractures in China from 2012 to 2014: a National Survey of 512,187 individuals.

The China National Fracture Study has been conducted to provide a national dataset of traumatic fractures across China. A national representative sample of 512,187 individuals was selected. The population-weighted incidence rates, distribution, injury mechanisms, and risk factors for traumatic fractures were identified for various groups of individuals. INTRODUCTION: The China National Fracture Study (CNFS) has been conducted to provide a comprehensive and up-to-date national dataset of traumatic fractures across China. This study aims to report the national incidences and distributions of traumatic fractures that occurred in 2012, 2013, and 2014 and to analyze the risk factors. METHODS: A national representative sample of individuals was selected from 24 rural counties and 24 urban cities of 8 provinces using stratified random sampling and the probability proportional to size (PPS) methodology. Participants were interviewed to identify whether they sustained traumatic fractures of the trunk and/or four extremities that had occurred in 2012, 2013, and 2014. The main risk factors associated with traumatic fractures were analyzed by multiple logistic regression models. RESULTS: A total of 512,187 individuals, including 259,649 males and 252,538 females, participated in the CNFS. The population-weighted incidence rates of traumatic fractures in China were calculated to be 2.5 (95% CI, 2.2-2.8) per 1000 population in 2012, 2.8 (95% CI, 2.5-3.3) in 2013, and 3.2% (95% CI, 2.8-3.6) in 2014. The population-weighted incidence rates of fragility fractures among participants aged 65 years and older were calculated to be 27.4 (95% CI, 21.4-33.4) per 1000 population in 2012, 36.0 (95% CI, 28.6-43.5) in 2013, and 42.4 (95% CI, 34.9-49.9) in 2014. The most common cause of fracture was low-energy injuries, followed by traffic accidents. For all age groups, sleeping less than 7 h was a risk factor for traumatic fractures. Alcohol consumption and previous fracture history were identified as risk factors for adults aged 15 years and over. Cigarette smoking was found to be a risk factor for males aged 15-64 years old. For individuals aged 15-64 years old, underweight incurred a risk effect for males and overweight for females. Alcohol consumption, sleeping less than 7 h per day, living in the central and eastern regions, a body mass index less of than 18.5, and having a previous fracture history were identified as strong risk factors for fragility fractures. CONCLUSION: The national incidence, distribution, and injury mechanisms for traumatic fractures were revealed in the CNFS. Risk factors were identified for various groups of individuals.

Diagnostic accuracy of deep learning in orthopaedic fractures: a systematic review and meta-analysis.

AIM: To gather and compare related clinical studies, and to investigate the accuracy and reliability of deep learning in detecting orthopaedic fractures. MATERIALS AND METHODS: This study is a retrospective combination and interpretation of prospectively acquired data. Articles from PubMed, EMBASE, the Cochrane library databases, and reference lists of the qualified articles were retrieved. Heterogeneity between studies was assessed using a random effective model. Pooled sensitivity, specificity, diagnostic odds ratio, and area under the receiver operating characteristic curve (AUC) were obtained by a random model. This work was managed from October 2018 to March 2020. RESULTS: Fourteen studies were included in this systematic review and nine were synthesized in the meta-analysis. The pooled sensitivity and specificity for the whole group (17 trials, 5,434 images) were 0.87 and 0.91, respectively. The AUC was 0.95. Eight trials (1,574 images) were included in the long-bone group, which contained seven studies. The pooled sensitivity was 0.96 and specificity was 0.94. The AUC was 0.99. Heterogeneity existed in the four pooled results of the whole group and the pooled specificity of the long-bone group. CONCLUSIONS: Deep learning is reliable in fracture diagnosis and has high diagnostic accuracy, which is similar to that of general physicians and is unlikely to produce a large number of false diagnoses; however, the ability of deep learning to localize the fracture needs more attention and testing. Deep learning can be extremely helpful with pre-classification of clinical diagnoses.

Provision of Lipid-Based Nutrient Supplements to Mothers During Pregnancy and 6 Months Postpartum and to Their Infants from 6 to 18 Months Promotes Infant Gut Microbiota Diversity at 18 Months of Age but Not Microbiota Maturation in a Rural Malawian Setting: Secondary Outcomes of a Randomized Trial.

BACKGROUND: Diet may alter the configuration of gut microbiota, but the impact of prenatal and postnatal nutritional interventions on infant gut microbiota has not been investigated. OBJECTIVE: We evaluated whether providing lipid-based nutrient supplements (LNSs) to mother-infant dyads promotes a more diverse and mature infant gut microbiota, compared to maternal supplementation with multiple micronutrients (MMN) or iron and folic acid (IFA). METHODS: We enrolled 869 pregnant women in a randomized trial in Malawi. There were 3 study groups, with women receiving 1 MMN capsule daily during pregnancy and 6 mo postpartum, or 1 LNS sachet (20 g) daily during pregnancy and 6 mo postpartum, or 1 IFA capsule daily (during pregnancy) then a placebo daily (postpartum). Infants in the LNS group received LNS from 6 to 18 mo; infants in the other groups did not receive supplements. The infants' fecal microbiota were characterized by PCR amplification and sequencing of the bacterial 16S rRNA gene (variable region 4). The primary outcomes were microbiota α diversity and maturation [as microbiota-for-age z score (MAZ)]. Specific associations of taxa with intervention were established with indicator species analysis (ISA). RESULTS: Primary outcomes did not differ between IFA and MMN groups, so these groups were combined (IFA + MMN). Mean ± SD α diversity was higher in the LNS group at 18 mo for Shannon index [3.01 ± 0.57 (LNS) compared with 2.91 ± 0.60 (IFA + MMN), P = 0.032] and Pielou's evenness index [0.61 ± 0.08 (LNS) compared with 0.60 ± 0.09 (IFA + MMN), P = 0.043]; no significant differences were observed at 1, 6, 12, or 30 mo. MAZ and ß diversity did not differ at any age. We found 10 and 3 operational taxonomic units (OTUs) positively associated with LNS and IFA + MMN, respectively; however, these associations became nonsignificant following false discovery rate correction at 10%. CONCLUSIONS: Prenatal and postnatal LNS intake promoted infant gut microbiota diversity at 18 mo, after 12 mo of child supplementation, but did not alter microbiota maturation. This trial was registered at clinicaltrials.gov as NCT01239693.