Objective:Comprehensive characterization of B-cell phenotypes and spatial distribution in oral lichen planus (OLP) and related oral lichenoid lesions (OLL)(OLP/OLL), with an emphasis on transcriptomic profiling and functional analysis, to uncover the epigenetic mechanisms underlying B cell-mediated immune regulation within the oral mucosal microenvironment.Methods:Single-cell RNA sequencing raw data were sourced from the GSE211630 database, encompassing samples from 2 cases of erosive OLP (EOLP), 3 cases of non-erosive OLP (NEOLP) and 1 healthy control (NORMAL). Following stringent quality control, the data underwent normalization, selection of highly variable genes and batch effect correction. Subsequent analyses included dimensionality reduction and unsupervised clustering to identify distinct cell populations. This study collected pathological specimens from 3 OLP/OLL patients and 3 healthy controls who were treated at the Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to December 2023. Using 10X Genomics Visium HD spatial transcriptomics technology, tissue sections were processed through dewaxing, staining and histological imaging, enabling the reconstruction of nucleic acid structures and the capture of gene expression profiles. Data analysis included quality assessment, gene quantification, normalization, dimensionality reduction and clustering. Furthermore, cell type deconvolution was performed using the robust cell type decomposition algorithm, integrating single-cell transcriptomic data to accurately predict and spatially resolve cell type distributions within the tissue microenvironment.Results:After integrating single-cell data from EOLP, NEOLP and NORMAL, cells were classified into seven major categories: B/plasma cells, endothelial cells, epithelial cells, fibroblasts, myeloid cells, smooth muscle cells and T/natural killer cells. The proportion of B/plasma cells varied significantly among the three groups, accounting for 10.7% (1 693/15 815), 3.8% (833/21 653) and 0.4% (47/11 556) of the total cells respectively. Further clustering analysis of B/plasma cells identified four distinct subpopulations: naive B cells, activated B cells, memory B cells and plasma cells. In the EOLP group, these subpopulations constituted 25.9% (348/1 344), 45.9% (617/1 344), 3.3% (45/1 344) and 24.9% (334/1 344) of the B/plasma cells respectively. In the NEOLP group, they represented 31.6% (195/617), 59.6% (368/617), 0.2% (1/617) and 8.6% (53/617). Howerer, only plasma cells were detected in the NORMAL group. Spatial analysis revealed that B cells were actively involved in the formation of tertiary lymphoid structures (TLS) at various stages in OLP/OLL samples, with a prominent structural organization observed in secondary follicle-like TLS. Within these structures, the expressions of T cells marker gene CD3E and B cells marker gene MS4A1 were significantly elevated. Additionally, in secondary follicle-like TLS, the gene encoding follicular dendritic cell secreted protein, germinal center marker gene B cell lymphoma 6 and the gene for activation induced cytidine deaminase also showed strong expression. In OLP/OLL samples, plasma cell marker gene CD38, immunoglobulin (IGH) G3, IGHG1, IGHM, IGHD, IGHE, imunoglobulin Kappa constant, immunoglobulin alpha 1, immunoglobulin Lambda constant 1 and complement gene C3 all exhibited high levels of expression.Conclusions:Compared to normal mucosa, extensive B-cell infiltration is observed in both OLP and OLL, accompanied by significant differences in B-cell phenotypes and proportions. B cells appear to play a central role in local immune responses, primarily through the formation of TLS. However, the precise functional mechanisms underlying their involvement require further investigation.

Colonic mucosal healing is the ultimate goal of ulcerative colitis (UC) treatment, but it remains difficult to realize. Given the higher incidence of UC in males and the beneficial effect of estrogen on UC, we conducted this study to examine the therapeutic potential of estrogen receptor β (ERβ), the primary ER subtype in colon, on mucosal healing in UC. Our study is the first to report that ERβ activation degree was positively correlated with mucosal healing in patients with UC. Furthermore, ERβ activation enhanced mucosal healing in mice with dextran sulfate sodium-induced and biopsy-induced colonic injuries. Mechanistically, ERβ activation promoted autophagy of colonic epithelial cells by inhibiting branched-chain amino acid transport, leading to focal adhesion kinase (FAK) activation. Activated FAK promoted focal adhesion turnover and colonic epithelial cell migration, ultimately facilitating mucosal healing. ERβ ^-/- colitis mice exhibited impaired mucosal healing compared to wild-type littermates, highlighting the crucial effect of ERβ. Importantly, combination with ERβ-agonist diarylpropionitrile enhanced the amelioration of 5-aminosalicylic acid, a standard UC treatment agent, against mouse colitis. These findings attest to the crucial role of ERβ activation in colonic mucosal healing and may further inform the development of novel strategies for UC treatment.

Objective:Comprehensive characterization of B-cell phenotypes and spatial distribution in oral lichen planus (OLP) and related oral lichenoid lesions (OLL)(OLP/OLL), with an emphasis on transcriptomic profiling and functional analysis, to uncover the epigenetic mechanisms underlying B cell-mediated immune regulation within the oral mucosal microenvironment.Methods:Single-cell RNA sequencing raw data were sourced from the GSE211630 database, encompassing samples from 2 cases of erosive OLP (EOLP), 3 cases of non-erosive OLP (NEOLP) and 1 healthy control (NORMAL). Following stringent quality control, the data underwent normalization, selection of highly variable genes and batch effect correction. Subsequent analyses included dimensionality reduction and unsupervised clustering to identify distinct cell populations. This study collected pathological specimens from 3 OLP/OLL patients and 3 healthy controls who were treated at the Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to December 2023. Using 10X Genomics Visium HD spatial transcriptomics technology, tissue sections were processed through dewaxing, staining and histological imaging, enabling the reconstruction of nucleic acid structures and the capture of gene expression profiles. Data analysis included quality assessment, gene quantification, normalization, dimensionality reduction and clustering. Furthermore, cell type deconvolution was performed using the robust cell type decomposition algorithm, integrating single-cell transcriptomic data to accurately predict and spatially resolve cell type distributions within the tissue microenvironment.Results:After integrating single-cell data from EOLP, NEOLP and NORMAL, cells were classified into seven major categories: B/plasma cells, endothelial cells, epithelial cells, fibroblasts, myeloid cells, smooth muscle cells and T/natural killer cells. The proportion of B/plasma cells varied significantly among the three groups, accounting for 10.7% (1 693/15 815), 3.8% (833/21 653) and 0.4% (47/11 556) of the total cells respectively. Further clustering analysis of B/plasma cells identified four distinct subpopulations: naive B cells, activated B cells, memory B cells and plasma cells. In the EOLP group, these subpopulations constituted 25.9% (348/1 344), 45.9% (617/1 344), 3.3% (45/1 344) and 24.9% (334/1 344) of the B/plasma cells respectively. In the NEOLP group, they represented 31.6% (195/617), 59.6% (368/617), 0.2% (1/617) and 8.6% (53/617). Howerer, only plasma cells were detected in the NORMAL group. Spatial analysis revealed that B cells were actively involved in the formation of tertiary lymphoid structures (TLS) at various stages in OLP/OLL samples, with a prominent structural organization observed in secondary follicle-like TLS. Within these structures, the expressions of T cells marker gene CD3E and B cells marker gene MS4A1 were significantly elevated. Additionally, in secondary follicle-like TLS, the gene encoding follicular dendritic cell secreted protein, germinal center marker gene B cell lymphoma 6 and the gene for activation induced cytidine deaminase also showed strong expression. In OLP/OLL samples, plasma cell marker gene CD38, immunoglobulin (IGH) G3, IGHG1, IGHM, IGHD, IGHE, imunoglobulin Kappa constant, immunoglobulin alpha 1, immunoglobulin Lambda constant 1 and complement gene C3 all exhibited high levels of expression.Conclusions:Compared to normal mucosa, extensive B-cell infiltration is observed in both OLP and OLL, accompanied by significant differences in B-cell phenotypes and proportions. B cells appear to play a central role in local immune responses, primarily through the formation of TLS. However, the precise functional mechanisms underlying their involvement require further investigation.

Objective To investigate the distribution of the thalassemia genotypes and the characteristics of blood cell parameters in Changshou District,Chongqing.Methods Totally 4126 samples sent to our hospital were studied from June 2018 to March 2023.All samples were detected for thalassemia genotype and blood cells.The parameters of blood cells:redblood cell(RBC),hemoglobin(Hb),mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC),red blood cell distribution width CV(RDW-CV),red blood cell distribution width SD(RDW-SD)were detected.Gap polymerase chain reaction(Gap-PCR)combined with reverse dot blot hybridization were used to detect alpha and beta thalassemia genotype.The rate and distribution characteristics of thalassemia gene in Changshou district were analyzed.Results Among 4126 samples,408 cases of α and β thalassemia were detected,accounting for 9.89%.Among these,there were 255 α-thalassemia cases.-α3.7/αα was the most common genotype.Two cases of--αSEA/-α3.7 and one cases of--SEA/HKαα were also detected.There were 153 cases of β-thalassemia and CD17 accounted for the highest proportion.The date of MCV,MCH,MCHC in-α3.7/αα,--SEA/αα,-α4.2/αα and ααCS/αα groups was significantly difference compared with control group(P<0.05).Parameters of MCV and MCH in CD17,CD41-42 and Ivs-2-654 groups were lower than those in control group(P<0.05),while RBC,RDW-CV and RDW-SD were higher than those in control group,the difference was statistically significant.Conclusion The most common genotype in thalassemia were-α3.7/αα,--SEA/αα,-α4.2/αα,CD17,CD41-42 and Ivs-2-654 in Changshou District,Chongqing.The parameters of MCV,MCHC,MCH,Hb,RBC,RDW-CV and RDW-SD have important clinical significance for the screening of thalassemia.

Objective:To investigate the roles of secretory phosphoprotein 1(SPP1)in the progression of colorectal cancer(CRC)and the underlying mechanism.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)database was used to obtain the expression of SPP1 gene in CRC.Immunohistochemistry analysis and Western blotting were used to detect the expression of SPP1 in distal normal colorectal tissues,adjacent tissues,CRC tissues,normal colorectal cell lines and CRC cell lines.The cell viability,colony formation,migration and invasion of CRC cells as well as the activation of AKT/glycogen synthase kinase 3β(GSK3β)signaling pathway and the expression of epithelial-mesenchymal transition(EMT)-related proteins in HT-29 cells and HCT-116 cells were detected by CCK-8 assay,colony formation assay,trranswell assay and Western blotting after SPP1 knockdown in vitro through lentiviral infection carrying shRNA against SPP1 gene.Tumor formation assay was used to detect the effect of SPP1 knockdown on the growth and lung metastasis of transplanted HT-29 tumor in vivo.Results:SPP1 expression was significantly increased in CRC tissues and cell lines(P<0.001)and was associated with poor prognosis of CRC patients according to GEPIA database analysis.The expression of SPP1 protein was significantly upregulated in CRC tissues and cells(P<0.001).After knockdown of SPP1 expression,the cell viability,colony formation,migration and invasion of CRC cells were significantly decreased(P<0.001),the expression of phosphorylated AKT(phospho-AKT,p-AKT),phosphorylated GSK3β(phospho-GSK3β,p-GSK3β),Snail and Vementin were significantly decreased(P<0.001),while E-cadherin expression was significantly increased(P<0.001).Knockdown of SPP1 expression inhibited the growth and lung metastasis of HT-29 cell tumor xenografts in mice.Conclusion:SPP1 knockdown can inhibit the proliferation,migration and invasion of CRC cells,which may be related to the reduction of AKT/GSK3β signaling activity.

Objective:To investigate the roles of secretory phosphoprotein 1(SPP1)in the progression of colorectal cancer(CRC)and the underlying mechanism.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)database was used to obtain the expression of SPP1 gene in CRC.Immunohistochemistry analysis and Western blotting were used to detect the expression of SPP1 in distal normal colorectal tissues,adjacent tissues,CRC tissues,normal colorectal cell lines and CRC cell lines.The cell viability,colony formation,migration and invasion of CRC cells as well as the activation of AKT/glycogen synthase kinase 3β(GSK3β)signaling pathway and the expression of epithelial-mesenchymal transition(EMT)-related proteins in HT-29 cells and HCT-116 cells were detected by CCK-8 assay,colony formation assay,trranswell assay and Western blotting after SPP1 knockdown in vitro through lentiviral infection carrying shRNA against SPP1 gene.Tumor formation assay was used to detect the effect of SPP1 knockdown on the growth and lung metastasis of transplanted HT-29 tumor in vivo.Results:SPP1 expression was significantly increased in CRC tissues and cell lines(P<0.001)and was associated with poor prognosis of CRC patients according to GEPIA database analysis.The expression of SPP1 protein was significantly upregulated in CRC tissues and cells(P<0.001).After knockdown of SPP1 expression,the cell viability,colony formation,migration and invasion of CRC cells were significantly decreased(P<0.001),the expression of phosphorylated AKT(phospho-AKT,p-AKT),phosphorylated GSK3β(phospho-GSK3β,p-GSK3β),Snail and Vementin were significantly decreased(P<0.001),while E-cadherin expression was significantly increased(P<0.001).Knockdown of SPP1 expression inhibited the growth and lung metastasis of HT-29 cell tumor xenografts in mice.Conclusion:SPP1 knockdown can inhibit the proliferation,migration and invasion of CRC cells,which may be related to the reduction of AKT/GSK3β signaling activity.

Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis (RA), but the relationship between the two phenomena remains unclear. We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA. Cholinergic function and protein citrullination levels in patients with RA and collagen-induced arthritis (CIA) mice were collected. In both neuron-macrophage coculture system and CIA mice, the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases (PADs) was assessed by immunofluorescence. The key transcription factors for PAD4 expression were predicted and validated. Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues. The cholinergic or alpha7 nicotinic acetylcholine receptor (α7nAChR) deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo, respectively. Especially, the activation deficiency of α7nAChR induced the earlier onset and aggravation of CIA. Furthermore, deactivation of α7nAChR increased the expression of PAD4 and specificity protein-3 (SP3) in vitro and in vivo. Our results suggest that cholinergic dysfunction-induced deficient α7nAChR activation, which induces the expression of SP3 and its downstream molecule PAD4, accelerating protein citrullination and the development of RA.

Background Mitochondrial dynamin-related protein 1 (DRP1) regulates mitochondrial division and plays an important role in maintaining hepatocyte function. However, the role of DRP1 in cadmium exposure-induced maternal liver damage in pregnant mice remains unclear. Objective To investigate the role and mechanism of DRP1 in maternal liver damage induced by cadmium exposure during pregnancy. Methods This study consisted of animal experiments and cell experiments. (1) Animal experiments. Mice at 14 days of gestation were randomly divided into three groups: a control group, a low-dose cadmium group (LCd group: 2.5 mg·kg⁻¹), and a high-dose cadmium group (HCd group: 5 mg·kg⁻¹). The pregnant mice were intraperitoneally injected with cadmium chloride (CdCl₂) for 6 and 24 h in the next morning. The weights of pregnant mice, uterus, maternal liver, and fetal mice were recorded after sacrifice. Serum and liver of pregnant mice were collected, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected, and liver tissues were stained with HE to observe changes in liver function and liver tissue structure. The expressions of oxidative phosphorylation-related proteins, hypoxia inducible factor-1α (HIF-1α) and DRP1 proteins in liver of pregnant mice were detected by Western blotting. (2) Cell experiments. AML12 cells were treated with CdCl₂ (10 μmol·L⁻¹) for 0, 2, 6, 12, and 24 h. The expressions of oxidative phosphorylation-related proteins, DRP1, and hypoxia inducible factor-1α (HIF-1α) proteins were detected. AML12 cells were pretreated with DRP1 inhibitor Mdivi-1 for 1 h and then CdCl₂ (10 μmol·L⁻¹) for 12 h to detect the expression of oxidative phosphorylation-related proteins and DRP1 protein. AML12 cells were treated with Hif-1α siRNA for 48 h and CdCl₂ (10 μmol·L⁻¹) for 6 h to detect the expression of HIF-1α and DRP1 proteins. Results The results of animal experiments showed that cadmium exposure in pregnant mice had no effects on maternal liver weight and liver coefficient. However, the histomorphological changes and necrosis in hepatocytes were observed. Compared with the control group, the serum ALT and AST levels of pregnant mice in the LCd group were significantly increased after 6 h (P＜0.05), and the levels in the HCd group were significantly increased after 6 and 24 h (P＜0.05). Cadmium exposure during pregnancy significantly up-regulated HIF-1α and DRP1 expressions and down-regulated the expressions of oxidative phosphorylation-related proteins in maternal livers. In vitro cell experiments showed that the expressions of oxidative phosphorylation-related proteins was significantly decreased and HIF-1α and DRP1 protein expressions were significantly increased in the AML12 cells treated with CdCl₂ for 6 h. Mdivi-1 pretreatment significantly antagonized the inhibitory effect of cadmium on the expressions of oxidative phosphorylation-related proteins in AML12 cells, while Hif-1α siRNA pretreatment significantly antagonized the up-regulative effect of cadmium on DRP1 expression in AML12 cells. Conclusion Cadmium exposure in pregnant mice may up-regulate DRP1 expression by activating HIF-1α signaling, then inhibit oxidative phosphorylation level of hepatic cells, and ultimately lead to maternal liver damage.

Objective:To analyze the risk factors for complications of the retromandibular approach in patients with parotid gland posterior and lower pole tumors.Methods:A retrospective analysis was conducted on the clinical data of 140 patients with parotid posterior lower pole tumors admitted to the Xingtai Third Hospital from October 2019 to October 2021. They were divided into two groups based on whether complications occurred: the occurrence group and the non occurrence group. General data of the two groups of patients were collected, including age, gender, course of disease, previous surgical history, number of tumors, tumor length, resection range, facial nerve dissociation, tumor site resection frequency, and fascia preservation; Single factor and logistic multivariate analysis were conducted to determine the risk factors for complications of the posterior retromandibular approach in patients with parotid gland posterior and lower pole tumors.Results:A total of 140 patients with parotid gland posterior lower pole tumors underwent retromandibular approach treatment, with complications occurring in 38 cases (27.14%), including 7 cases of temporary facial paralysis, 10 cases of facial depression, 11 cases of Frey syndrome, 2 cases of fistula, and 8 cases of sensory abnormalities of the greater auricular nerve. Through logistic multivariate analysis, it was found that the number of tumors ≥ 2 ( OR=2.856), the resection range (total resection) ( OR=2.477), the number of surgeries ≥3 ( OR=5.637), facial nerve dissociation ( OR=3.526), and lack of fascia preservation ( OR=2.551) were all risk factors for postoperative complications in patients with parotid posterior pole tumors (all P<0.05). Conclusions:In clinical practice, relevant prevention and treatment measures should be formulated for these high-risk factors to reduce the incidence of postoperative complications.

Objective:To investigate the clinical characteristics and short-term follow-up outcomes of primary nephrotic syndrome (PNS) children infected with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the Omicron variant outbreak in Shanghai, and to provide a reliable reference for clinicians in the diagnosis and treatment.Methods:It was a case-control study. The clinical data of children with PNS (PNS group) who were diagnosed and followed-up up to 1 year in the nephrology department of four children's medical centers in Shanghai, and the children (control group) who had no underlying diseases and were infected with SARS-CoV-2 in Shanghai Jinshan Public Health Center, including the data when they were infected with SARS-CoV-2, were retrospectively analyzed.Results:(1) From March 30th to April 13th, 2022, 6 PNS children in Shanghai were infected with SARS-CoV-2, including 5 boys and 1 girl. The median age was 4.5 (2.0, 11.0) years old. And 30 children were matched by sex, age and disease type as control group, including 20 males and 10 females. The median age was 4.5 (2.0, 9.0) years. There were no significant differences between the PNS group and the control group in clinical symptoms (including fever duration), treatment regimens, vaccine doses and virus clearance time (all P>0.05). (2) The 6 children with PNS included 3 cases of steroid-sensitive type, 3 cases of steroid-resistant type, 2 cases of minimal change disease, 2 cases of focal segmental glomerulosclerosis and 2 cases with no renal biopsy. Before SARS-CoV-2 infection, their primary disease-PNS were stable, and urine protein was negative, four of them were under maintenance treatment with oral steroids or immunosuppressive drugs. At the time of SARS-CoV-2 infection, the symptoms of all of the 6 cases were mild, no severe, critical or fatal cases, and they were all cured and discharged from hospital through medical isolation observation or symptomatic treatment of infections. (3) Five cases of them still had discomfort symptoms such as cough, anorexia, and fatigue after being discharged from the hospital, which lasted for about 1 week. Within 1 year of follow-up, none of the children have suffered from "recurrent positive PCR results" or "secondary infection" of the SARS-CoV-2. (4) Among them, 4 cases of PNS relapsed after SARS-CoV-2 infection, timely addition of steroids was effective, their urine protein quickly turned negative, and there was no recurrence after 1 year of follow-up. (5) Before infection with SARS-CoV-2, the levels of immunoglobulin IgG were lower than the normal reference value in the 4 cases with PNS recurrence. Conclusions:During the Omicron variant outbreak in Shanghai, the infection of SARS-CoV-2 in children with PNS are resulted in high transmission among household contacts. Most of them have mild symptoms and good prognosis. PNS is prone to relapse after SARS-CoV-2 infection, and steroid therapy is effective and safe for these relapse. IgG may be a potential marker for the prognosis of PNS children infected with SARS-CoV-2.