[摘 要] 目的：探讨lncRNA SOX2-OT是否通过调控miR-215-5p/NIN1/RPN12结合蛋白1同源物（NOB1）信号通路抑制结直肠癌（CRC）HCT-116细胞增殖和迁移。方法：收集2022年6月至2024年5月期间武汉市金银潭医院收治的29例CRC患者的癌及癌旁组织标本，以及CRC细胞SW480、HCT-116、HP116和LoVo及人结肠上皮细胞HCoEpiC。qPCR法检测CRC组织和细胞中SOX2-OT、miR-215-5p和NOB1 mRNA的表达水平。利用RNA干扰技术分别将敲低或过表达SOX2-OT质粒及阴性对照质粒转染至HCT-116细胞，实验分为对照组、si-NC组、si-SOX2-OT组、si-SOX2-OT + inhibitor（Inh）NC组、si-SOX2-OT + miR-215-5p Inh组、si-SOX2-OT + oe-NC组、si-SOX2-OT + oe-NOB1组。qPCR法检测各组细胞中SOX2-OT、miR-215-5p和NOB1 mRNA表达水平，通过MTT、划痕愈合实验、Transwell实验和流式细胞术分别检测各组细胞的增殖、迁移、侵袭及凋亡水平，WB法检测各组细胞中E-cadherin、N-cadherin、vimentin、Bcl-2、BAX、PCNA、MMP-9和NOB1蛋白表达水平。通过双萤光素酶报告基因实验验证miR-215-5p与SOX2-OT和NOB1之间的靶向关系。结果：CRC组织和细胞中SOX2-OT和NOB1 mRNA均呈高表达、miR-215-5p低表达（均P < 0.05）。敲低SOX2-OT表达的HCT-116细胞中SOX2-OT和NOB1 mRNA表达水平、增殖、划痕愈合率和侵袭细胞数量，以及细胞中N-cadherin、vimentin、Bcl-2、NOB1、PCNA和MMP-9蛋白表达均显著降低（均P < 0.05），miR-215-5p、细胞凋亡率、E-cadherin和BAX蛋白表达均显著升高（均P < 0.05）。敲低miR-215-5p表达或上调NOB1表达均可减弱敲低SOX2-OT表达对细胞的抑制作用（均P < 0.05）。miR-215-5p与SOX2-OT和NOB之间存在靶向关系。结论：敲低SOX2-OT表达可促进miR-215-5p表达，抑制NOB1表达进而抑制HCT-116细胞的增殖、迁移、侵袭并促进细胞凋亡。

OBJECTIVE To investigate the effects of metformin （Met） on liver injury in non-alcoholic steatohepatitis （NASH） rats by regulating the PI3K/AKT/PDGF signaling pathway. METHODS NASH model was constructed by feeding rats with a high- glucose and high-fat diet， and assigned into Model group， Met low-dose group （Met-L group， 100 mg/kg）， Met medium-dose group （Met-M group， 200 mg/kg）， Met high-dose group （Met-H group， 400 mg/kg）， and high dose of Met+PI3K activator group （Met-H+740 Y-P group， 400 mg/kg Met+50 mg/kg 740 Y-P）， with 12 rats in each group. Another 12 rats were regarded as the Control group. Each group of rats was orally administered/injected with the corresponding medication once a day for 6 consecutive weeks. The changes in body weight and liver index of rats were recorded and analyzed. The pathological damage [evaluation of non-alcoholic fatty liver disease activity score （NAS）]， lipid deposition （calculation of the proportion of oil red O-positive staining area）， and fibrosis （calculation of collagen deposition score） were observed in liver tissue of rats. The levels of inflammatory factors [interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α）] in serum and liver tissue， the levels of serum lipid metabolism indicators [total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）] and liver function indicators [aspartate aminotransferase （AST） and alanine Δ 基金项目 武汉市知识创新专项项目（No.2022020801010588）； aminotransferase （ALT）] were measured. The expression levels of PI3K/AKT/PDGF signaling pathway-related proteins and Caspase-3 in liver tissue of rats were determined. RESULTS Compared with the Control group， body weight， liver index， the levels of serum lipid metabolism indicators and liver function indicators， the levels of IL-6 and TNF-α in serum and liver tissue， the NAS， the proportion of oil red O-positive staining area， the collagen deposition fraction， and the levels of phosphorylated PI3K and AKT proteins， as well as the expression levels of PDGF and Caspase-3 proteins in liver tissue， were all significantly increased （P＜0.05）. The liver tissue showed severe pathological damage， characterized by an abundance of lipid droplets and pronounced collagen deposition. After the intervention with Met， the aforementioned quantitative indicators and pathological changes in rats were significantly improved in a dose- dependent manner （P＜0.05）. 740 Y-P could reverse the improvement effects of high dose of Met on the above indexes of rats （P＜ 0.05）. CONCLUSIONS Met can improve liver damage， and alleviate inflammatory reactions and liver fibrosis of NASH rats， the mechanism of which may be associated with inhibiting PI3K/AKT/PDGF signaling pathway.

ObjectiveTo compare the hepatic bile acid profile between a mouse model of metabolic-associated steatohepatitis （MASH） induced by a high-fat， high-sugar， and high-cholesterol diet combined with intraperitoneal injection of 10% carbon tetrachloride （CCl₄） and MASH cases in clinical practice， and to investigate the feasibility of this model in studying drug interventions on bile acid profile in MASH. MethodsA total of 30 male C57BL/6J mice were randomly divided into control group and model group， with 15 mice in each group. The mice in the control group were given normal diet and drinking water and weekly injections of olive oil， and those in the model group were given a high-fat， high-sugar， and high-cholesterol diet， high-sugar drinking water， and weekly injections of CCl₄+olive oil. At the end of weeks 8， 12， and 16， 5 mice were selected from each group to collect samples. Behavioral assessments were performed， and body weight and liver wet weight were measured； liver pathology and lipid deposition were evaluated by HE staining， SAF scoring， oil Red O staining， the semi-quantitative analysis of stained area， the serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， and liver triglyceride （TG） content； Sirius Red staining was performed for liver tissue to assess liver fibrosis； ultra-performance liquid chromatography-tandem mass spectrometry and targeted metabolomics were used to measure the hepatic bile acid profile， including cholic acid （CA）， glycocholic acid （GCA）， chenodeoxycholic acid （CDCA）， glycochenodeoxycholic acid （GCDCA）， ursodeoxycholic acid （UDCA）， tauroursodeoxycholic acid （TUDCA）， hyodeoxycholic acid （HDCA）， and glycodeoxycholic acid （GDCA）. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsCompared with the control group at the same time point， the model group had disheveled and dull fur， reduced activity， and relatively slow reactions at weeks 8， 12， and 16， as well as significant increases in liver wet weight （P<0.05）， the serum level of ALT （P<0.05）， the content of TG in the liver （P<0.05）， and SAF score （P<0.05）. As for the differentially expressed bile acids in liver tissue， compared with the control group at week 8， the model group had significantly higher levels of CA and CDCA and significantly lower levels of UDCA， TUDCA， HDCA， and GDCA （all P<0.05）； compared with the control group at week 12， the model group had significantly higher levels of CA， GCA， CDCA， and GCDCA and significantly lower levels of UDCA and HDCA （all P<0.05）； compared with the control group at week 16， the model group had significantly higher levels of CA， GCA， CDCA， GCDCA， and TUDCA and significantly lower levels of UDCA， HDCA， and GDCA （all P<0.05）. As for the differentially expressed bile acids in the bile acid pool of liver tissue， compared with the control group at week 8， the model group had significantly higher levels of CA and CDCA and significantly lower levels of UDCA， TUDCA， GDCA， and HDCA （all P<0.05）； compared with the control group at weeks 12 and 16， the model group had significantly higher levels of GCA and GCDCA and significantly lower levels of UDCA， GDCA， and HDCA （all P<0.05）. ConclusionThere are significant changes in the hepatic bile acid profile in a mouse model of MASH induced by a high-fat， high-sugar， and high-cholesterol diet combined with CCl₄， which are similar to the changes in bile acids in MASH cases in clinical practice， suggesting that this model can be used to explore the interventional effect of drugs on the bile acid profile in MASH.

Objective:To investigate the clinical effects of adjustable external fixation traction combined with arthroscopic microfracture in the treatment of osteochondral lesions of the talus (OLT).Methods:A retrospective study was conducted to analyze the data of 27 OLT patients who had been treated at Department of Orthopedics, Beijing Rehabilitation Hospital from May 2017 to March 2022. There were 16 males and 11 females, aged (32.4±7.2) years. Lesion site: 23 medial and 4 lateral cases; Hepple staging: 7 cases at stage Ⅰ, 15 cases at stage Ⅱ, and 5 cases at stage Ⅲ; disease duration: (10.6±3.3) months. All the patients were treated by adjustable external fixation traction combined with arthroscopic microfracture. Recorded were the patients' visual analogue scale (VAS) pain scores and American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores at 6 months and 12 months after surgery, levels of interleukin-1 (IL-1), interleukin-6 (IL-6) and tumour necrosis factor- α (TNF- α) at 1 month after surgery, lesion area at 12 months after surgery, and incidence of complications. Results:The follow-up time for this cohort was (16.2±6.7) months. The AOFAS score was (61.52±6.75) points before surgery, (84.15±5.56) points at 6 months after surgery and (95.67±4.30) points at 12 months after surgery. The VAS score was (5.88±1.02) points before surgery, (2.12±0.48) points at 6 months after surgery and (0.66±0.36) points at 12 months after surgery. The two-by-two comparisons between the 3 time points for the above items were statistically significant ( P<0.05). IL-1 was (32.37±6.64) pg/mL, IL-6 (34.04±7.12) pg/mL, and TNF- α (17.89±4.96) ng/L at 1 month after surgery in the 27 patients, all of which were significantly lower than their preoperative levels [(96.63±14.80) pg/mL, (102.33±20.42) pg/mL, and (54.48±9.33) ng/L] ( P<0.05). The lesion area was (28.66±6.52) mm 2 at 12 months after surgery, significantly smaller than the value before surgery [(128.52±11.32) mm 2] ( P<0.05). Infection at the adjustable external fixation needle track occurred in 1 patient and lower limb thrombosis in 2 patients. Conclusion:In the treatment of OLT, adjustable external fixation and traction combined with arthroscopic microfracture can achieve satisfactory results and improve symptoms for the patients.

Objective To investigate the sleep structure of Parkinson's disease (PD) patients with rapid eye movement sleep behavior disorder (RBD) and its correlations with cognitive function, depressive state and motor function. Methods A total of 120 PD patients were enrolled in this study, and divided into PD+RBD group (n=45) and PD group (n=75) based on the presence or absence of RBD. Sleep structure was recorded for both groups. The scores of the Montreal Cognitive Assessment (MoCA), Hamilton Anxiety Scale (HAMA), Hamilton Depression Scale (HAMD), Unified Parkinson's Disease Rating Scale Part Ⅲ (UPDRS-Ⅲ) and Scales for Outcomes in Parkinson's Disease-Autonomic (SCOPA-AUT) were compared between the two groups. The correlations of MoCA, HAMA, HAMD, UPDRS-Ⅲ and SCOPA-AUT scores with sleep structure in the PD+RBD group were analyzed. Results The PD+RBD group showed significantly higher percentage of non-rapideye movement (NREM) at stage 1, arousal index, periodic limb movements during sleep (PLMS) and apnea-hypopnea index (AHI) compared to the PD group, while the percentages of NREM at stage 3, rapid eye movement (REM) sleep, sleep efficiency and lowest oxygen saturation were significantly lower (P < 0.05). The PD+RBD group also had significantly higher HAMA, HAMD, UPDRS-Ⅲ and SCOPA-AUT scores, and significantly lower MoCA scores compared to the PD group (P < 0.05). In the PD+RBD group, the percentages of NREM at stage 1, arousal index, PLMS and AHI were positively correlated with HAMA, HAMD, UPDRS-Ⅲ and SCOPA-AUT scores, and negatively correlated with MoCA scores (P < 0.05). Conversely, the percentages of NREM at stage 3, REM sleep, sleep efficiency and lowest oxygen saturation were negatively correlated with HAMA, HAMD, UPDRS-Ⅲ and SCOPA-AUT scores, and positively correlated with MoCA scores (P < 0.05). Conclusion PD patients with RBD exhibit disrupted sleep structure, which is closely associated with cognitive function, depressive state, motor function, and autonomic nervous system function.

Objective:To analyze the status of respiratory pathogen detection and the clinical features in children with Mycoplasma pneumoniae pneumonia (MPP). Methods:A prospective, multicenter study was conducted to collect clinical data, including medical history, laboratory examinations and multiplex PCR tests of children diagnosed with MPP from 4 hospitals in China between November 15 th and December 20 th, 2023. The multiplex PCR results and clinical characteristics of MPP children in different regions were analyzed. The children were divided into severe and mild groups according to the severity of the disease. Patients in the severe group were further divided into Mycoplasma pneumoniae (MP) alone and Multi-pathogen co-detection groups based on whether other pathogens were detected besides MP, to analyze the influence of respiratory pathogen co-detection rate on the severity of the disease. Mann-Whitney rank sum test and Chi-square test were used to compare data between independent groups. Results:A total of 298 children, 136 males and 162 females, were enrolled in this study, including 204 children in the severe group with an onset age of 7.0 (6.0, 8.0) years, and 94 children in the mild group with an onset age of 6.5 (4.0, 7.8) years. The level of C-reactive protein, D-dimer, lactic dehydrogenase (LDH) were significantly higher (10.0 (5.0, 18.0) vs. 5.0 (5.0, 7.5) mg/L, 0.6 (0.4, 1.1) vs. 0.5 (0.3, 0.6) mg/L, 337 (286, 431) vs. 314 (271, 393) U/L, Z=2.02, 2.50, 3.05, all P<0.05), and the length of hospitalization was significantly longer in the severe group compared with those in mild group (6.0 (6.0, 7.0) vs. 5.0 (4.0, 6.0) d, Z=4.37, P<0.05). The time from onset to admission in severe MPP children was significantly shorter than that in mild MPP children (6.0 (5.0, 9.5) vs. 9.0 (7.0, 13.0) d, Z=2.23, P=0.026). All patients completed the multiplex PCR test, with 142 cases (47.7%) MPP children detected with 21 pathogens including adenovirus 25 cases (8.4%), human coronavirus 23 cases (7.7%), rhinovirus 21 cases (7.0%), Streptococcus pneumoniae 21 cases (7.0%), influenza A virus 18 cases (6.0%). The pathogens with the highest detection rates in Tianjin, Shanghai, Wenzhou and Chengdu were Staphylococcus aureus at 10.7% (8/75), adenovirus at 13.0% (10/77), adenovirus at 15.3% (9/59), and both rhinovirus and Haemophilus influenzae at 11.5% (10/87) each. The multi-pathogen co-detection rate in severe MPP children was significantly higher than that in mild MPP group (52.9% (108/204) vs. 36.2% (34/94), χ2=10.62, P=0.005). Among severe MPP children, there are 89 cases in the multi-pathogen co-detection group and 73 cases in the simple MPP group. The levels of LDH, D-dimer and neutrophil counts in the multi-pathogen co-detection group were significantly higher than those in the simple MPP group (348 (284, 422) vs. 307 (270, 358) U/L, 0.8 (0.5, 1.5) vs. 0.6 (0.4, 1.0) mg/L, 4.99 (3.66, 6.89)×10 9vs. 4.06 (2.91, 5.65)×10 9/L, Z=5.17, 4.99, 6.11, all P<0.05). Conclusions:The co-detection rate of respiratory pathogens, LDH and D-dimer in children with severe MPP were higher than those with mild MPP. Among severe MPP children the stress response of children in co-detection group was more serious than that of children with simple MPP.

Objective To investigate the safety and feasibility of peripheral vascular intervention via the ipsilateral transulnar access(TUA)due to failure of transradial access(TRA)puncture.Methods The clinical data of 2546 peripheral vascular interventions via TRA,which were performed at authors'hospital between January 2019 and December 2021,were retrospectively analyzed.Among the 2546 interventions,TRA puncture failed in 37 procedures,and in 27 of these patients the ipsilateral TUA puncture had to be adopted.The puncture success rate,surgical success rate and puncture approach-related complications of TUA of the 27 patients receiving ipsilateral TUA puncture were analyzed.Results The success rate of ipsilateral TUA puncture after TRA puncture failed was 96.3％(26/27),and in one patient transfemoral access(TFA)puncture had to be substituted because of the ulnar artery spasm.The total success rate of interventional procedures was 96.3％(26/27).No serious complications occurred,and the incidence of minor complications was 19.2％(5/26).Conclusion Preliminary results indicate that for the experienced TRA operators,using ipsilateral TUA puncture due to failure of TRA puncture is a safe and feasible strategy choice.

Breast cancer is the most common cancer in women and one of the deadliest cancers worldwide.According to the distribution of tumor tissue,breast cancer can be divided into invasive and non-invasive forms.The cancer cells in invasive breast cancer pass through the breast and through the immune system or systemic circulation to different parts of the body,forming metastatic breast cancer.Drug resistance and distant metastasis are the main causes of death from breast cancer.Research on breast cancer has attracted extensive attention from researchers.In vitro construction of tumor models by tissue engineering methods is a common tool for studying cancer mechanisms and anticancer drug screening.The tumor microenvironment consists of cancer cells and various types of stromal cells,including fibroblasts,endothelial cells,mesenchymal cells,and immune cells embedded in the extracellular matrix.The extracellular matrix contains fibrin proteins(such as types Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅵ,and Ⅹcollagen and elastin)and glycoproteins(such as proteoglycan,laminin,and fibronectin),which are involved in cell signaling and binding of growth factors.The current traditional two-dimensional(2D)tumor models are limited by the growth environment and often cannot accurately reproduce the heterogeneity and complexity of tumor tissues in vivo.Therefore,in recent years,research on three-dimensional(3D)tumor models has gradually increased,especially 3D bioprinting models with high precision and repeatability.Compared with a 2D model,the 3D environment can better simulate the complex extracellular matrix components and structures in the tumor microenvironment.Three-dimensional models are often used as a bridge between 2D cellular level experiments and animal experiments.Acellular matrix,gelatin,sodium alginate,and other natural materials are widely used in the construction of tumor models because of their excellent biocompatibility and non-immune rejection.Here,we review various natural scaffold materials and construction methods involved in 3D tissue-engineered tumor models,as a reference for research in the field of breast cancer.

Objective To observe the effects of alum borneol nanoemulsion on the secretion of inflammatory factors by lipopolysac-charide(LPS)induced mononuclear phagocyte(RAW264.7)in mice.Methods Using LPS to induce RAW264.7 as an M1 pheno-type,the changes of cell morphological were observed under microscope.The effects of different concentrations of alum borneol nanoemul-sion on RAW264.7 cell viability was detected by methylthiazolyldiphenyl-tetrazolium bromide(MTT)colorimetry.The relative mRNA expression of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and interleukin-10(IL-10)were detectd by real-time quantitative polymerase chain reaction(RT-PCR).The positive expression rates of M1/M2 polarized surface markers CD68 and CD206 were measured by flow cytometry The contents of IL-6,TNF-α and IL-10 in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Results The morphology of RAW264.7 cells changed obviously when cultured with LPS and alum bor-neol nanoemulsion in vitro.Low,medium,and high concentrations(8.15,16.30,32.60mg/ml)of alum borneol nanoemulsion had no toxic effects on RAW264.7.Compared with the blank group without any treatment,the expression of CD68 was higher(P＜0.001),the expression of CD206 was lower(P＞0.05)in the model group after 24hours of LPS stimulation of RAW264.7 cells with good growth,in-dicating that M1 macrophage modeling was successfully.After drug intervention,compared with the model group,the expression of CD68membrane surface molecules was lower than before(P＜0.001),and CD206 was higher than before(P＜0.001),indicating that alum borneol nanoemulsion could increase the expression of M2macrophage membrane protein.Compared with the model group(LPS group),alum borneol nanoemulsion could inhibit the secretion of TNF-α and IL-6 in LPS-induced macrophage inflammation model(P＜0.01,P＜0.05),but promoted the secretion of IL-10(P＜0.001).Compared with the model group(LPS group),the results of RT-qPCR showed that alum borneol nanoemulsion significantly inhibited the expression of TNF-α and IL-6mRNA(P＜0.01,P＜0.05),but promoted the expression of IL-10mRNA(P＜0.01).Conclusion The alum borneol nanoemulsionhas has good anti-in-flammatory activity on LPS-induced inflammatory responses in RAW264.7 cells,which may be achieved by inhibiting the secretion and expression of inflammatory cytokines.

Objective:To recheck the Histoplasma capsulatum and Coccidioides sp. strains of the past by molecular identification and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS). Methods:The phylogenetic relationships among the ten Histoplasma capsulatum isolates and reference strains were analyzed by multilocus sequence typing (MLST). Based on the Coi region, Coccidioides posadasii was distinguished from Coccidioides immitis accurately. MALDI-TOF MS was used to set up the MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides sp. for rapid identification. In addition, hierarchical clustering of spectra was compared with MLST. Results:An unrooted dendrogram constructed with MLST showed that ten individuals of Histoplasma capsulatum were divided into three clades: Eurasia clade, Australia clade and North American class 2 clade, in agreement with the establishment by MALDI-TOF MS cluster analysis. All individuals of Coccidioides sp. were identified as Coccidioides posadasii with Coi region primers. The in-house MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides posadasii was expanded and reached an identified accuracy of 100%. Conclusions:We improve the recognition of Histoplasma capsulatum and Coccidioides posadasii by molecular pathways which shows the major species or clades in Chinese mainland. The in-house MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides posadasii can provide a new efficient way to identify those pathogens rapidly.