ObjectiveTo investigate the mechanisms by which the combination of berberine （BBR） and baicalin （BAI） ameliorates type 2 diabetes mellitus （T2DM） due to internal accumulation of dampness-heat from the perspectives of gut microbiota and metabolomics. MethodsAntibiotics were used to induce pseudo-sterile mice. Thirty pseudo-sterile mice were randomized into a normal fecal microbiota transplantation group （n=10） and a T2DM （syndrome of internal accumulation of dampness-heat） fecal microbiota transplantation group （n=20）. The mice were then administrated with suspensions of fecal microbiota from healthy volunteers and a patient with T2DM due to internal accumulation of dampness-heat by gavage， respectively. Each mouse received 200 µL suspension every other day for a total of 15 times to reshape the gut microbiota. The T2DM model mice were then assigned into a model group （n=8） and a BBR-BAI group （n=11）. BBR was administrated at a dose of 200 mg·kg^-1， and BAI was administrated in a ratio of BBR-BAI 10∶1 based on preliminary research findings. The administration lasted for 8 consecutive weeks. Fasting blood glucose （FBG）， glycated hemoglobin （HbA1c）， insulin （INS）， triglycerides （TG）， total cholesterol （CHOL）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） levels were measured to evaluate the effects of the BBR-BAI combination on glucose and lipid metabolism and liver function in T2DM mice. Hematoxylin-eosin staining was employed to observe pathological changes in the colon tissue. The expression of claudin-1， zonula occludens-1 （ZO-1）， and occludin in the colon tissue was determined by Western blot. Real-time quantitative polymerase chain reaction（Real-time PCR） was employed to assess the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the colon tissue. The fecal microbiota composition and differential metabolites were analyzed by 16S rRNA sequencing and ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry （UPLC-Q-TOF-MS）， respectively. ResultsThe BBR-BAI combination lowered the FBG， HbA1c， and INS levels （P<0.05， P<0.01） and alleviated insulin resistance （P<0.01） in T2DM mice. Additionally， BBR-BAI elevated the levels of ZO-1， occludin， and claudin-1 （P<0.05， P<0.01） and down-regulated the expression levels of TNF-α， IL-1β， and IL-6 in the colon （P<0.05， P<0.01）. The results of 16S rRNA sequencing showed that BBR-BAI increased the relative abundance of Ligilactobacillus， Phascolarctobacterium， and Akkermansia （P<0.05）， while significantly decreasing the relative abundance of Alistipes， Odoribacter， and Colidextribacter （P<0.05）. UPLC-Q-TOF-MS identified 28 differential metabolites， which were primarily involved in arachidonic acid metabolism and α-linolenic acid metabolism. ConclusionBBR-BAI can ameliorate T2DM due to internal accumulation of dampness-heat by modulating the relative abundance of various bacterial genera in the gut microbiota and the expression of fecal metabolites.

Background Staff in biosafety laboratories (BSL) are more likely to experience occupational burnout and other psychological issues due to their unique working environment and high job demands. However, current research in this field tends to focus on overall analyses, overlooking the internal differences within this group. Objective To explore latent profiles of occupational burnout among BSL workers and their influencing factors, providing a reference for targeted burnout interventions. Methods In 2022, cluster random sampling was used to select 8023 BSL workers in Xinjiang. Data were collected via a questionnaire survey, including a self-designed general information questionnaire, the Maslach Burnout Inventory-General Survey (MBI-GS), and the Effort-Reward Imbalance (ERI) scale, to analyze demographic characteristics, occupational burnout, and job stress status. After data cleaning, latent profile analysis (LPA) was applied to identify burnout profiles, followed by K-means clustering for verification. Logistic regression was used to analyze influencing factors with burnout profiles as the dependent variable. Results After excluding invalid questionnaires, 7924 valid questionnaires were retained, with an recovery rate of 98.77%. The LPA results indicated that occupational burnout among BSL staff was categorized into three latent profiles: a low burnout profile (2208 individuals, 27.86%), a moderate burnout profile (4636 individuals, 58.51%), and a high burnout profile (1080 individuals, 13.63%). A similar grouping was derived from K-means clustering analysis (28.71%, 53.37%, and 17.92%), and a Sankey diagram demonstrated significant overlap between the two classification results. Statistically significant differences were observed across the three latent profiles in all three dimensions of burnout: emotional exhaustion, cynicism, and reduced personal accomplishment. The final logistic regression model revealed that the influencing factors exhibited characteristics of partial overlap and partial differentiation. Among the common influencing factors, the shared protective factors included: age ≥50 years (OR_{moderate burnout}=0.42, OR_{high burnout}=0.45), monthly income of 4000-6000 RMB (OR_{moderate burnout}=0.83, OR_{high burnout}=0.71), good sleep quality (OR_{moderate burnout}=0.70, OR_{high burnout}=0.13) and very good sleep quality (OR_{moderate burnout}=0.51, OR_{high burnout}=0.12), and engaging in physical exercise ≥5 times per week (OR_{moderate burnout}=0.74, OR_{high burnout}=0.48). The protective effects were more pronounced for the high burnout profile. The common risk factors included: 21–30 years of service (OR_{moderate burnout}=1.38, OR_{high burnout}=3.00), ≥31 years of service (OR_{moderate burnout}=1.47, OR_{high burnout}=3.39), working in "other positions" (OR_{moderate burnout}=1.46, OR_{high burnout}=1.56), and occupational stress (ERI>1) (OR_{moderate burnout}=2.62, OR_{high burnout}=3.25), with the risk effects being more significant for the high burnout profile. Regarding differentiated influencing factors, protective factors exclusively associated with the moderate burnout profile included having an associate degree (OR=0.70) and engaging in physical exercise 3-4 times per week (OR=0.74). For the high burnout profile, exclusive protective factors included age 40-50 years (OR=0.56), while exclusive risk factors included having a senior professional title (OR=1.85), holding a managerial role (OR=1.78), a high number of night shifts per month (OR=2.66), and a very high number of night shifts per month (OR=3.93). Conclusion BSL workers exhibit three distinct latent burnout profiles. Occupational burnout requires greater attention, and targeted prevention and intervention strategies should be developed based on worker subtypes.

Specific pathogen-free(SPF)chickens are widely used in the research of avian diseases and vaccines.Vertically transmissible diseases are transmitted to chickens through vertical transmission,seriously affecting their survival rate,increasing production costs,and causing significant economic losses to the poultry industry,while severely impacting the breeding and use of SPF chickens.Therefore,it is crucial for researchers and managers to enhance their understanding of vertically transmissible pathogens in chickens and to develop effective monitoring measures.Quality monitoring is an important part of ensuring the quality of SPF chickens,with pathogen detection being the primary step.Based on this,it is necessary to cultivate qualified SPF chickens through purification methods and biosecurity measures.This paper reviews the major vertically transmissible pathogens in chickens,including viral pathogens,bacterial pathogens and mycoplasmas,as well as their detection methods.This study compares the differences in microbiological testing items and methods for SPF chickens between the U.S.corporate standard and the Chinese national standard.Analysis of the results shows that in both standards,vertically transmissible pathogens such as Escherichia coli,Proteus mirabilis,Salmonella,and avian leukosis are not included in the microbiological testing items for SPF chickens.Instead,these pathogens are characterized by mixed infections,and outbreaks can seriously affect flock health.To produce higher-quality SPF chickens,it is necessary to include these pathogens in the mandatory testing items.The aim of this paper is to help readers understand the relevant standards for microbiological monitoring of SPF chickens,the hazards of vertically transmissible pathogens,and prevention and control strategies,so as to provide a reference for the detection and purification of pathogens in SPF chickens.

The lip plays a crucial role in facial aesthetics,making them a focal point of interest for orthodontists and patients through-out orthodontic treatment due to their three-dimensional changes in morphology and position.The degree of improvement in lip morphol-ogy significantly impacts post-treatment satisfaction among orthodontic patients.Therefore,understanding the influencing factors of lip morphology and its correlation with orthodontic interventions enables orthodontists to predict post-treatment changes in lip morphology more accurately,facilitating the customization of orthodontic treatment plans.In this paper,we reviewed the progress of methods for evaluating lip morphology and its influencing factors in order to provide reference for clinical practice.

Successful pregnancy in placental mammals substantially depends on the establishment of maternal immune tolerance to the semi-allogenic fetus.Disorders in this process are tightly asso-ciated with adverse pregnancy outcomes including recurrent miscarriage (RM).However,an in-depth understanding of the systematic and decidual immune environment in RM remains largely lacking.In this study,we utilized single-cell RNA-sequencing (scRNA-seq) to comparably analyze the cellular and molecular signatures of decidual and peripheral leukocytes in normal and unex-plained RM pregnancies at the early stage of gestation.Integrative analysis identifies 22 distinct cell clusters in total,and a dramatic difference in leukocyte subsets and molecular properties in RM cases is revealed.Specifically,the cytotoxic properties of CD8+ effector T cells,nature killer(NK),and mucosal-associated invariant T (MAIT) cells in peripheral blood indicates apparently enhanced pro-inflammatory status,and the population proportions and ligand-receptor interac-tions of the decidual leukocyte subsets demonstrate preferential immune activation in RM patients.The molecular features,spatial distribution,and the developmental trajectories of five decidual NK(dNK) subsets have been elaborately illustrated.In RM patients,a dNK subset that supports embryonic growth is diminished in proportion,while the ratio of another dNK subset with cyto-toxic and immune-active signature is significantly increased.Notably,a unique pro-inflammatory CD56 + CD16 + dNK subset substantially accumulates in RM decidua.These findings reveal a com-prehensive cellular and molecular atlas of decidual and peripheral leukocytes in human early pregnancy and provide an in-depth insight into the immune pathogenesis for early pregnancy loss.

Objective:To determine the expression of microRNA-188-5p (miR-188-5p) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to assess the effect of its downregulation on the proliferation and invasion of CSCC cells.Methods:From November 2012 to October 2018, 50 surgically resected CSCC tissue specimens and 50 paracancerous normal skin tissue specimens were collected from the First Affiliated Hospital of Xinxiang Medical College in Henan Province. Real-time fluorescence-based quantitative PCR (qPCR) was employed to determine the expression of miR-188-5p in CSCC tissues, paracancerous normal skin tissues, CSCC cell lines SCL-1, A431 and HSC-5, and a human immortalized keratinocyte line HaCaT. Cultured A431 and HSC-5 cells were both divided into 2 groups: miR-188-5p inhibitor group and negative control group, which were transfected with a miR-188-5p inhibitor and its negative control respectively. Then, qPCR was performed to determine the relative expression level of miR-188-5p (expressed as 2 -△△Ct), and cell counting kit-8 (CCK8) and Transwell assays were conducted to assess cellular proliferative activity and invasive ability respectively in the above groups. Dual-luciferase reporter assay was performed to investigate interactions between miR-188-5p and phosphatase and tensin homologue deleted on chromosome 10 (PTEN), and Western blot analysis to determine the protein expression of PTEN, total Akt (t-Akt) and phosphorylated Akt (p-Akt). Two independent samples were compared by using t test. Results:The relative expression level of miR-188-5p was significantly higher in the CSCC tissues (5.213 ± 3.138) than in the paracancerous normal skin tissues (1.010 ± 0.364, t = 9.187, P < 0.001), and significantly higher in the SCL-1, A431 and HSC-5 cells (3.858 ± 0.163, 7.068 ± 0.262 and 4.572 ± 0.413, respectively) than in the HaCaT cells (1.079 ± 0.300, t = 17.890, 21.110 and 8.737, respectively, all P < 0.05). Compared with the negative control group, the miR-188-5p inhibitor group showed significantly decreased miR-188-5p expression in both A431 and HSC-5 cells (both P < 0.01), and decreased proliferative activity and invasive ability of both A431 and HSC-5 cells (all P < 0.05). Dual-luciferase reporter assay showed that the downregulation of miR-188-5p significantly increased the expression of PTEN, but inhibited the expression of p-Akt in A431 and HSC-5 cells. Conclusion:MiR-188-5p is highly expressed in CSCC tissues and cells, and the downregulation of miR-188-5p may inhibit the proliferative activity and invasive ability of CSCC cells by regulating the PTEN/Akt pathway.

Objective To evaluate the effect of downregulation of microRNA (miR)-373 expression on cell cycle and apoptosis of a cutaneous squamous cell carcinoma (CSCC) cell line A431.Methods A431 cells at exponential growth phase were classified into 3 groups:miR-373 inhibitor group and negative control group transfected with miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.At 48 hours after the transfection,real-time PCR was performed to determine the expression of miR-373 in the above 3 groups,cell counting kit-8 (CCK-8) assay to evaluate the effect of downregulated expression of miR-373 on the proliferation of A431 cells,flow cytometry to investigate the distribution of cell cycle and changes in apoptosis of A431 cells in different treatment groups,and colorimetric analysis to detect the changes in caspase-3 activity in different treatment groups.Statistical analysis was carried out with SPSS 17.0 software by using two-sample t test for the comparison between two groups,one-way analysis of variance (ANOVA) for the comparison among 3 groups,and least significant difference (LSD)-t test for multiple comparisons.Results The expression of miR-373 was significantly lower in the miR-373 inhibitor group (0.120 ± 0.036) than in the untreated group (1.002 ± 0.022) and negative control group (1.037 ± 0.028,LSD-t =36.21,34.83,respectively,both P ＜ 0.001).At 48,72 and 96 hours,the miR-373 inhibitor group showed significantly decreased proliferative activity of A375 cells compared with the untreated group and negative control group (F =10.805,13.720 and 30.907 respectively,P =0.038,0.010 and 0.001 respectively).The proportion of A375 cells in G0/G1 phase was significantly higher in the miR-373 inhibitor group (64.69％ ± 1.18％) than in the untreated group (52.74％ ± 0.66％,t =15.51,P ＜ 0.001) and negative control group (53.80％ ± 0.80％,t =13.24,P ＜ 0.001).The proportion of total apoptotic cells and activity of caspase-3 in the miR-373 inhibitor group were 22.69％ ± 1.24％ and 1.238 ± 0.057 respectively,which were significantly higher than those in the untreated group (9.62％ ± 1.14％,0.413 ± 0.028 respectively,both P ＜ 0.001)and negative control group (9.66％ ± 0.97％,0.437 ± 0.036 respectively,both P ＜ 0.001).Conclusion MiR-373 may play an important role in the regulation of cell cycle and induction of apoptosis of the CSCC cell line A431.

The expression or dysfunction of long non-coding RNAs (lncRNAs) is closely related to various hereditary diseases, autoimmune diseases, metabolic diseases and tumors. LncRNAs were also recently recognized as functional regulators of fibrosis, which is a secondary process in many of these diseases and a primary pathology in fibrosis diseases. We review the latest findings on lncRNAs in fibrosis diseases of the liver, myocardium, kidney, lung and peritoneum. We also discuss the potential of disease-related lncRNAs as therapeutic targets for the clinical treatment of human fibrosis diseases.
Autoimmune Diseases ; Fibrosis ; Genetic Diseases, Inborn ; Humans ; Kidney ; Liver ; Lung ; Metabolic Diseases ; Myocardium ; Pathology ; Peritoneum ; RNA, Long Noncoding

Objective To investigate the expression of microRNA-373 (miR-373) in cutaneous squamous cell carcinoma (CSCC) tissues and cells,and to explore its effects on cell invasion.Methods Real-time PCR was performed to determine the expression of miR-373 in CSCC tissues and paralesional normal skin tissues,as well as in CSCC cell lines (A431 and SCL-1) and HaCaT cells.A431 cells were divided into 4 groups:miR-373 mimic group,miR-373 inhibitor group and negative control group which were transfected with miR-373 mimic,miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.Cell invasion assay was performed to evaluate effects of miR-373 downregulation on cell invasion.Western blot analysis was conducted to assess effects of miR-373 downregulation on the protein expression of matrix metalloproteinase-2 (MMP-2) and MMP-9.Results Expression of miR-373 was significantly higher in the CSCC tissues (2.465 ± 0.218) than in the paralesional normal skin tissues (1.000 ± 0.000,P ＜ 0.05),and higher in SCL-1 cells (1.864 ± 0.178) and A431 cells (2.919 ± 0.277) than in HaCaT cells (1.000 ± 0.000,P ＜ 0.05).Most notably,miR-373 expression was also markedly higher in metastatic CSCC tissues than in non-metastatic CSCC tissues (3.323 ± 0.344 vs.1.914 ± 0.161,t =4.158,P =0.000 4).Compared with the untreated group and negative control group,the miR-373 mimic group showed significantly increased miR-373 expression and invasive ability,while the miR-373 inhibitor group showed markedly decreased miR-373 expression and invasive ability (all P ＜ 0.05).Conclusion MiR-373 downregulation can significantly suppress the invasion of A431 cells,and obviously decrease the protein expression of MMP-2 and MMP-9.

Objective To evaluate effects of downregulation of glucose?6?phosphate dehydrogenase(G6PD) expression on proliferation and cell cycle distribution of cutaneous squamous cell carcinoma(CSCC)cells. Methods Western blot analysis was performed to measure the protein expression of G6PD in normally cultured human HaCaT keratinocytes, SCL?1 and A431 CSCC cells. When A431 cells grew to 85%-90%confluence, a small interfering RNA (siRNA)targeting G6PD(G6PD?siRNA group)and a negative control siRNA(siRNA control group)were transfected into them separately, and untransfected A431 cells served as the untransfected group. CCK?8 assay was performed to evaluate proliferative activity of the A431 cells on days 0, 1, 2, 3 and 4 after transfection, Western blot analysis to measure G6PD, cyclin D1 and CDK4 protein expressions in A431 cells, and flow cytometry to analyze cell cycle distribution in A431 cells after 48 hours of additional culture. Results The protein expression of G6PD was significantly higher in normally cultured SCL?1 cells(0.308 ± 0.023)and A431 cells(0.643 ± 0.046)than in HaCaT cells(0.100 ± 0.019, both P<0.05), and significantly higher in A431 cells than in SCL?1 cells(P<0.05). The G6PD?siRNA group showed significantly decreased protein expressions of G6PD, cyclin D1 and CDK4(0.134 ± 0.027, 0.154 ± 0.017 and 0.166 ± 0.017, respectively)compared with the untransfected group(0.425 ± 0.029, 0.344 ± 0.024 and 0.330 ± 0.020 respectively)and siRNA control group(0.444 ± 0.033, 0.350 ± 0.027 and 0.348 ± 0.018 respectively) (all P<0.05). Besides, the G6PD?siRNA group showed significantly decreased cellular proliferative activity on days 1-4 compared with the siRNA control group and untransfected group(all P<0.001), while there were no significant differences between the untransfected group and siRNA control group at any of the time points (all P > 0.05). Compared with the untransfected group and siRNA control group, the G6PD?siRNA group showed significantly higher proportions of A431 cells in G0/G1 phase(both P < 0.001), but significantly lower proportions of A431 cells in S phase(both P<0.001). Conclusion G6PD may play important roles in the regulation of proliferation and cell cycle distribution of CSCC cells.