Purpose To investigate the relationship between FN1 expression and clinical and pathologic features of laryngeal squamous cell carcinoma(LSCC)and the expression of tumor-associated macrophages(TAMs).Methods LSCC datasets GSE33232 and GSE84957 were analyzed and screened the differentially expressed gene FN1,and draw the receiver operating characteristic(ROC)curve.Bioinformatics analysis of FN1 expression,and prognosis in LSCC was performed.To investigate the effect of down-regulating FN1 expression in TU177 cells on the malignant bio-logical behavior of LSCC,we performed a scratch wound healing assay and a Transwell chamber assay to assess the effect of FN1 on cell proliferation,migration,and invasion in vitro.Immunohistochemical(IHC)staining was per-formed to detect the expression of FN1 and CD 163 in LSCC tissues.Results Analysis of the GSE33232 and GSE84957 datasets and online databases showed that FN1 was significantly overexpressed in LSCC tissues(P＜0.05),and patients with high FN1 expression had a significantly lower recurrence-free survival rate(HR=1.6,P=0.017).After transfection with si-FN1,the expression of FN1 in TU177 cells was significantly reduced(0.34±0.02 vs 1.00±0.03,P＜0.01).Compared with the control group,the down-regulation of FN1 expression inhibited the in vitro migra-tion(56.1±3.1 vs 19.23±1.0)and invasion(480±23 vs 288±20)ability of TU177 cells(both P＜0.01).Im-munohistochemistry findings showed that FN1 was highly expressed in both the tumor parenchyma(nest)and stromal cells of LSCC tissue,with a statistically significant difference[52.1％(24/46)vs 71.7％(33/46),P＜0.001].It was found that high expression of N-FN1 was associated with patients' pathological grade and lymph node metastasis(P＜0.05),while high expression of S-FN1 was associated with patients' age,lymph node metastasis,and TNM stage(P＜0.05).In addition,the co-expression of FN1 and CD163 was correlated with patients' pathological grad-ing,lymph node metastasis,and TNM stage(all P＜0.05).Conclusion FN1 and CD163 exhibit high expression levels in LSCC patients,which are closely associated with malignant progression,including invasion and metastasis.Notably,during LSCC progression,there may be a potential synergistic interaction between FN1 and CD 163-positive macrophages in the tumor microenvironment.

Coronavirus disease 2019 (COVID-19) is a multi-system disease that can lead to various severe complications. Acute limb ischemia (ALI) has been increasingly recognized as a COVID-19-associated complication that often predicts a poor prognosis. However, the pathophysiology and molecular mechanisms underlying COVID-19-associated ALI remain poorly understood. Hypercoagulability and thrombosis are considered important mechanisms, but we also emphasize the roles of vasospasm, hypoxia, and acidosis in the pathogenesis of the disease. The angiotensin-converting enzyme 2 (ACE2) pathway, inflammation, and platelet activation may be important molecular mechanisms underlying these pathological changes induced by COVID-19. Furthermore, we discuss the hypotheses of risk factors for COVID-19-associated ALI from genetic, age, and gender perspectives based on our analysis of molecular mechanisms. Additionally, we summarize therapeutic approaches such as use of the interleukin-6 (IL-6) blocker tocilizumab, calcium channel blockers, and angiotensin-converting enzyme inhibitors, providing insights for the future treatment of coronavirus-associated limb ischemic diseases.
Humans ; COVID-19/physiopathology* ; Ischemia/etiology* ; SARS-CoV-2 ; Extremities/blood supply* ; Risk Factors ; Interleukin-6/antagonists & inhibitors* ; Acute Disease ; Angiotensin-Converting Enzyme 2

Objective:To explore the characteristics of T cells and natural killer (NK) cells in cerebrospinal fluid of patients with multiple sclerosis (MS).Methods:Cerebrospinal fluids from patients with relapsing-remitting multiple sclerosis (RRMS) and healthy controls attending the Second Hospital of Lanzhou University from January 2023 to October 2024 were collected and analyzed by single-cell RNA sequencing (scRNA-seq) and flow cytometry, and T and NK cell characteristics were summarized and compared in the two groups. The proposed time-series analysis was used to explore the differentiation trajectories of T cells and NK cells.Results:A total of 3 patients with RRMS and 3 healthy controls underwent scRNA-seq, and 6 patients with RRMS and 4 healthy controls underwent flow cytometry. The composition of cerebrospinal fluid T and NK cell subtypes was similar in the RRMS group and the healthy control group, but the proportion of the cell subtypes differed. The RRMS group exhibited significantly higher frequencies of CD4 +Naive T- IL7R [1 529/9 055(16.89%) vs 1 423/9 910(14.36%), χ2=22.980, P<0.001], CD4 +Naive T- CCR7 [1 573/9 055(17.37%) vs 948/9 910(9.57%), χ2=250.114, P<0.001], and CD4 +Naive T- LTB [1 369/9 055(15.12%) vs 1 079/9 910(10.89%), χ2=75.336, P<0.001] subsets compared to controls. Conversely, the control group demonstrated greater proportions of CD4 +Th1- GZMA [1 255/9 910(12.66%) vs 719/9 055(7.94%), χ2=113.213, P<0.001] and CD8 +Tem- GZMK cells [1 607/9 910(16.22%) vs 1 232/9 055(13.61%), χ2=25.326, P<0.001] than the RRMS group. The transcription factors and gene expression of each T cell subtype were also different between the 2 groups, and CD4 initial T cells and CD8 effector T cells were located at the beginning and the end of the differentiation trajectory, respectively. Conclusions:The cerebrospinal fluid of MS patients is characterized by increased expression of genes involved in T cell differentiation and over-activation of immune cells.

Purpose To investigate the relationship between FN1 expression and clinical and pathologic features of laryngeal squamous cell carcinoma(LSCC)and the expression of tumor-associated macrophages(TAMs).Methods LSCC datasets GSE33232 and GSE84957 were analyzed and screened the differentially expressed gene FN1,and draw the receiver operating characteristic(ROC)curve.Bioinformatics analysis of FN1 expression,and prognosis in LSCC was performed.To investigate the effect of down-regulating FN1 expression in TU177 cells on the malignant bio-logical behavior of LSCC,we performed a scratch wound healing assay and a Transwell chamber assay to assess the effect of FN1 on cell proliferation,migration,and invasion in vitro.Immunohistochemical(IHC)staining was per-formed to detect the expression of FN1 and CD 163 in LSCC tissues.Results Analysis of the GSE33232 and GSE84957 datasets and online databases showed that FN1 was significantly overexpressed in LSCC tissues(P＜0.05),and patients with high FN1 expression had a significantly lower recurrence-free survival rate(HR=1.6,P=0.017).After transfection with si-FN1,the expression of FN1 in TU177 cells was significantly reduced(0.34±0.02 vs 1.00±0.03,P＜0.01).Compared with the control group,the down-regulation of FN1 expression inhibited the in vitro migra-tion(56.1±3.1 vs 19.23±1.0)and invasion(480±23 vs 288±20)ability of TU177 cells(both P＜0.01).Im-munohistochemistry findings showed that FN1 was highly expressed in both the tumor parenchyma(nest)and stromal cells of LSCC tissue,with a statistically significant difference[52.1％(24/46)vs 71.7％(33/46),P＜0.001].It was found that high expression of N-FN1 was associated with patients' pathological grade and lymph node metastasis(P＜0.05),while high expression of S-FN1 was associated with patients' age,lymph node metastasis,and TNM stage(P＜0.05).In addition,the co-expression of FN1 and CD163 was correlated with patients' pathological grad-ing,lymph node metastasis,and TNM stage(all P＜0.05).Conclusion FN1 and CD163 exhibit high expression levels in LSCC patients,which are closely associated with malignant progression,including invasion and metastasis.Notably,during LSCC progression,there may be a potential synergistic interaction between FN1 and CD 163-positive macrophages in the tumor microenvironment.

Objective:To explore the characteristics of T cells and natural killer (NK) cells in cerebrospinal fluid of patients with multiple sclerosis (MS).Methods:Cerebrospinal fluids from patients with relapsing-remitting multiple sclerosis (RRMS) and healthy controls attending the Second Hospital of Lanzhou University from January 2023 to October 2024 were collected and analyzed by single-cell RNA sequencing (scRNA-seq) and flow cytometry, and T and NK cell characteristics were summarized and compared in the two groups. The proposed time-series analysis was used to explore the differentiation trajectories of T cells and NK cells.Results:A total of 3 patients with RRMS and 3 healthy controls underwent scRNA-seq, and 6 patients with RRMS and 4 healthy controls underwent flow cytometry. The composition of cerebrospinal fluid T and NK cell subtypes was similar in the RRMS group and the healthy control group, but the proportion of the cell subtypes differed. The RRMS group exhibited significantly higher frequencies of CD4 +Naive T- IL7R [1 529/9 055(16.89%) vs 1 423/9 910(14.36%), χ2=22.980, P<0.001], CD4 +Naive T- CCR7 [1 573/9 055(17.37%) vs 948/9 910(9.57%), χ2=250.114, P<0.001], and CD4 +Naive T- LTB [1 369/9 055(15.12%) vs 1 079/9 910(10.89%), χ2=75.336, P<0.001] subsets compared to controls. Conversely, the control group demonstrated greater proportions of CD4 +Th1- GZMA [1 255/9 910(12.66%) vs 719/9 055(7.94%), χ2=113.213, P<0.001] and CD8 +Tem- GZMK cells [1 607/9 910(16.22%) vs 1 232/9 055(13.61%), χ2=25.326, P<0.001] than the RRMS group. The transcription factors and gene expression of each T cell subtype were also different between the 2 groups, and CD4 initial T cells and CD8 effector T cells were located at the beginning and the end of the differentiation trajectory, respectively. Conclusions:The cerebrospinal fluid of MS patients is characterized by increased expression of genes involved in T cell differentiation and over-activation of immune cells.

Acral melanoma(AM)is the most common histologic subtype of melanoma in dark-skinned patients and is associated with a worse prognosis and a high mortality rate,largely due to the inconspicuous nature of early-stage lesions,which can lead to late diagnosis.Because of the overlapping clinical and histopathological features of AM with other forms of cutaneous melanomas,early detection of AM requires a multidisciplinary approach that integrates various diagnostic modalities,including clinical examination,dermoscopy,histopathology,molecular testing,radiological imaging,and blood tests.While surgery is the preferred method of treatment for AM,other therapeutic options may be employed based on the stage and underlying etiology of the disease.Immune checkpoint inhibitors,molecular targeted therapy,radiotherapy,chemotherapy,and oncolytic virotherapy represent promising advanced treatment options for AM.In this review,we provide an overview of the latest advancements in diagnostic and therapeutic methods for AM,highlighting the importance of early detection and the prompt,individualized management of this challenging disease.

[摘 要] 目的：探究SOX9通过上调长链非编码RNA LINC01503的表达对喉鳞状细胞癌（LSCC）细胞的增殖、迁移、侵袭及肿瘤干细胞干性的影响。方法： 常规培养人LSCC细胞AMC-HN-8、TU177、TU212和TU686，用转染试剂将敲减序列及其对照核酸（si-SOX9-NC、si-SOX9#1、 si-SOX9#2、si-LINC01503-NC、si-LINC01503#1、si-LINC01503#2）或过表达质粒及其对照核酸（pcDNA3.1-SOX-NC、pcDNA3.1-SOX-oe、pcDNA3.1-LIN01503-NC和pcDNA3.1-LIN01503-oe）分别转染至TU177细胞或TU686细胞，记为si-SOX9-NC组、si-SOX9#1组、si-SOX9#2组、si-LINC01503-NC组、si-LINC01503#1组、si-LINC01503#2组；pcDNA3.1-SOX9-NC组、pcDNA3.1-SOX9-oe组、pcDNA3.1-LINC01503-NC组、pcDNA3.1-LINC01503-oe组、si-SOX9-NC + pcDNA3.1-LINC01503-NC组和si-SOX9 + pcDNA3.1-LINC01503-oe组。qPCR法检测SOX9 mRNA和LINC01503 在各组细胞中的表达，生物信息学分析SOX9与LINC0503启动子区的结合位点，双萤光素酶报告基因实验和染色质免疫共沉淀实验验证SOX9与LINC01503启动子区是否直接结合，WB法检测SOX9的敲减效率及LINC01503对TU177和TU686细胞干性标志物表达的影响，MTS法检测各组细胞的增殖活力，划痕愈合和Transwell小室实验检测各组细胞的迁移能力，克隆形成实验检测各组细胞的克隆形成能力。结果：SOX9在各种LSCC细胞中呈高表达（均P < 0.05），数据库数据分析显示，在头颈部鳞状细胞癌中，SOX9与LINC01503表达呈正相关（R = 0.12，P = 0.005 9）；SOX9可与LINC01503启动子区直接结合并促进其转录表达（均P < 0.05）；敲减LINC01503可明显抑制TU177细胞的增殖、迁移、侵袭（均P < 0.05），过表达LINC01503明显促进TU686细胞增殖、迁移、侵袭的能力（均P < 0.05），提高TU686细胞克隆形成能力和细胞干性标志物分子CD133、OCT4、SOX2的mRNA和蛋白水平表达（均P < 0.05），敲减LINC01503则均可抑制TU686细胞的克隆形成和细胞干性标志物的表达（均P < 0.05）；敲减SOX9均可明显抑制TU177细胞的增殖、迁移和侵袭能力，降低其干性细胞标志物的表达（均P < 0.05），同时过表达LINC01503则可部分逆转敲减SOX9对TU177细胞恶性生物学行为和干性标志物表达的抑制作用（均P < 0.05）。结论：SOX9和LINC01503在LSCC细胞中呈高表达，SOX9可能通过上调LINC01503表达提高LSCC细胞增殖、转移和侵袭能力和肿瘤干细胞干性。

Lennox-Gastaut syndrome(LGS)is a severe,epileptic encephalopathy.In recent years,a variety of drugs have been approved for the treatment of LGS.The U.S.Food and Drug Administration ap-proved clobazam and cannabidiol as adjunctive therapy for LGS in October 2011 and June 2018,respectively.This article provides an overview of clobazam and cannabidiol,including their chemical structures,pharmaco-logical actions,curative effects,safety profile,drug interactions,to introduce the current state of research and the achievements of both drugs.

Objective:To investigate the relationship between histone deacetylase (HDAC) gene polymorphism and type 2 diabetes mellitus (T2DM) in Bai and Han populations in Dali of Yunnan province.Methods:A total of 148 patients with T2DM of Bai and Han nationalities who received treatment in Dali Bai Autonomous Prefecture People's Hospital from May 2019 to March 2021 were included in the T2DM group. An additional 100 healthy controls of Bai and Han nationalities who concurrently received physical examination in the same hospital from May 2019 to December 2020 were included in the normal control group. The susceptibility genes of T2DM were detected using the Taqman MGB probe method. The susceptibility gene loci were amplified using polymerase chain reaction. The whole sequence of susceptibility gene was sequenced.Results:There were no significant differences in the distribution frequencies of rs2530223 genotype, rs11741808 genotype, rs2547547 genotype, and rs1741981 genotype between Bai and Han populations (all P > 0.05). There was a significant difference in blood lipid level between four loci ( t = -1.06, -0.19, 0.39, -2.12, -2.04, 0.16, 1.47, < 0.01, -0.16, -3.17, -2.93, 0.69, -2.58, -2.33, all P < 0.05). There was a significant difference in homeostasis model assessment of insulin resistance between different states (all P < 0.05). The frequency distributions of each genotype and each allele did not differ significantly between healthy control people of Bai nationality and T2DM patients of Bai nationality and between healthy control people of Han nationality and T2DM patients of Han nationality (all P > 0.05). Logistic regression analysis showed that the polymorphism was not an independent risk factor for T2DM. Conclusion:The relationships between HDAC gene polymorphism and T2DM, obesity and dyslipidemia differ between Bai and Han populations.

Hyponatremia is common in the attack of acute intermittent porphyria(AIP), which can cause epilepsy, coma and other adverse events and endanger the life of patients. Carbohydrate loading therapy is applied to control the attack of AIP in the clinic. But the application of glucose can exacerbate hyponatremia. It is difficult for clinicians to effectively correct hyponatremia while treating AIP with glucose. We reported a case of AIP whose refractory hyponatremia was corrected with short-term low-dose tolvaptan to improve knowledge in management.