[摘 要] 目的：探讨同源异型盒蛋白D11（HOXD11）对Ki-67的转录调控作用及其对喉鳞状细胞癌（LSCC）细胞恶性生物学行为的影响和机制。方法：结合高通量测序数据，通过GEO、UALCAN数据库分析HOXD11在LSCC中差异表达。收集2022年1月至2025年1月联勤保障部队第九八〇医院手术切除的60例LSCC患者的癌及癌旁组织标本，以及人LSCC细胞系AMC-HN-8、TU-177、TU-686和人正常喉上皮细胞（HNLEC），构建敲低或过表达HOXD11的细胞系，将细胞分为对照组、HOXD11敲低组及HOXD11过表达组。通过RT-qPCR法检测HOXD11和Ki-67基因在LSCC组织和细胞中mRNA表达水平，免疫组织化学（IHC）法分析两者在LSCC组织中的蛋白表达及分布，WB法进一步验证蛋白差异表达。采用MTS、克隆形成实验及Transwell实验分别检测敲低或过表达HOXD11对LSCC细胞增殖、迁移与侵袭的影响。通过双萤光素酶报告基因实验和染色质免疫沉淀（ChIP）实验验证HOXD11对Ki-67启动子活性的调控作用。结果：GEO和UALCAN数据库分析表明，HOXD11在LSCC中高表达（P < 0.01）。在LSCC组织中HOXD11和Ki-67 mRNA和蛋白表达均显著高于癌旁组织（均P < 0.01），同时，两者表达水平之间存在正相关（r = 0.26，P < 0.05）；LSCC细胞系中HOXD11 mRNA表达显著高于HNLEC（均P < 0.01）。敲低HOXD11显著抑制LSCC细胞的增殖、迁移和侵袭能力（均P < 0.01），而过表达HOXD11则促进细胞的这些恶性生物学行为（均P < 0.01）。双萤光素酶报告基因实验及ChIP实验均证实，HOXD11可直接结合到Ki-67启动子区上，调控其表达（P < 0.01）。回复实验显示，过表达Ki-67可部分逆转敲低HOXD11对LSCC细胞增殖、迁移与侵袭的抑制作用（均P < 0.01）。结论：HOXD11在LSCC组织及细胞系中均呈高表达，其机制在于通过直接调控Ki-67的转录活性，从而增强LSCC细胞的增殖、迁移及侵袭能力。

Objective To investigate the expression pattern of IQGAP2 in renal cell carcinoma tissues and cell lines,and to analyze its effects on the proliferation and migration of renal cell carcinoma cells.Methods Firstly,GEO database was used to screen differentially expressed genes between renal cell carcinoma tissues,and GEPIA,TIMER2.0 were used to analyze the expression level of IQGAP2 in renal cell carcinoma tissue.Subse-quently,knockdown(siRNA)and overexpression plasmids of IQGAP2 were constructed and transfected into ACHN and 786-O cell lines to perform a series of functional experiments to evaluate the effect of IQGAP2 on the malignant biological behavior of renal carcinoma cells.qRT-PCR and Western Blot were used to detect the expres-sion of EMT(epithelial-mesenchymal transition)related proteins after knockdown and overexpression of IQGAP2.Results In renal cell carcinoma tissues,the relative expression of IQGAP2 was significantly lower than in adja-cent normal tissues(P<0.001).Transfection of si-IQGAP2 in ACHN and 786-O cells effectively downregulated the mRNA and protein expression levels of IQGAP2(P<0.01),while transfection with an overexpression plasmid significantly upregulated its mRNA and protein expression(P<0.001).Further studies revealed that overexpression of IQGAP2 significantly inhibited the proliferation(P<0.05)and migration(P<0.01)of ACHN and 786-O cells,whereas knockdown of IQGAP2 enhanced their proliferation(P<0.05,P<0.001)and migration(P<0.01).Through qRT-PCR and Western blot analyses of EMT-related proteins,it was found that reduced expression of IQGAP2 promoted the epithelial-mesenchymal transition(EMT)process in renal cancer cells.Conclusions The expression of IQGAP2 is low in renal cell carcinoma tissues and cells,and the decrease of the expression level can promote the EMT process of renal cell carcinoma cells,and then enhance the proliferation and migration of renal cell carcinoma cells.IQGAP2 plays an important tumor suppressor role in renal cell carcinoma.

Objective To investigate the expression pattern of IQGAP2 in renal cell carcinoma tissues and cell lines,and to analyze its effects on the proliferation and migration of renal cell carcinoma cells.Methods Firstly,GEO database was used to screen differentially expressed genes between renal cell carcinoma tissues,and GEPIA,TIMER2.0 were used to analyze the expression level of IQGAP2 in renal cell carcinoma tissue.Subse-quently,knockdown(siRNA)and overexpression plasmids of IQGAP2 were constructed and transfected into ACHN and 786-O cell lines to perform a series of functional experiments to evaluate the effect of IQGAP2 on the malignant biological behavior of renal carcinoma cells.qRT-PCR and Western Blot were used to detect the expres-sion of EMT(epithelial-mesenchymal transition)related proteins after knockdown and overexpression of IQGAP2.Results In renal cell carcinoma tissues,the relative expression of IQGAP2 was significantly lower than in adja-cent normal tissues(P<0.001).Transfection of si-IQGAP2 in ACHN and 786-O cells effectively downregulated the mRNA and protein expression levels of IQGAP2(P<0.01),while transfection with an overexpression plasmid significantly upregulated its mRNA and protein expression(P<0.001).Further studies revealed that overexpression of IQGAP2 significantly inhibited the proliferation(P<0.05)and migration(P<0.01)of ACHN and 786-O cells,whereas knockdown of IQGAP2 enhanced their proliferation(P<0.05,P<0.001)and migration(P<0.01).Through qRT-PCR and Western blot analyses of EMT-related proteins,it was found that reduced expression of IQGAP2 promoted the epithelial-mesenchymal transition(EMT)process in renal cancer cells.Conclusions The expression of IQGAP2 is low in renal cell carcinoma tissues and cells,and the decrease of the expression level can promote the EMT process of renal cell carcinoma cells,and then enhance the proliferation and migration of renal cell carcinoma cells.IQGAP2 plays an important tumor suppressor role in renal cell carcinoma.

Objective:To explore the clinical value of a trinity strategy for the treatment of multiple orthopedic trauma.Methods:A retrospective case series study was conducted to analyze the clinical data of 1 267 patients with multiple orthopedic trauma admitted to Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine and the First Affiliated Hospital of Navy Medical University from June 2013 to May 2023, including 862 males and 405 females, aged 18-93 years [(55.2±19.8)years]. Associated injuries included hemorrhagic shock in 632 patients, traumatic wet lung in 274, cranial injuries in 135, abdominal and pelvic bleeding in 116, pneumothorax in 89, urinary injury in 13, and vesical rupture in 8. All the patients were treated with the trinity strategy and the treatment process was divided into the phases of first aid, remodeling, and rehabilitation. The first aid phase focused on stabilizing symptoms and saving lives; the remodeling phase centered on restoring the anatomical structure and alignment; the rehabilitation phase aimed for functional recovery through the integration of both Western and traditional Chinese medicine. The all-cause mortality within 30 days after surgery and fracture healing time were calculated; the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, Hospital for Special Surgery (HSS) knee score and the American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot score at the last follow-up and the overall excellent and good rate of all joint function scores were measured. The short form health survey (SF-36) scores were collected preoperatively and at 6 months postoperatively, including 8 aspects such as physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health. The incidence of postoperative complications was recorded.Results:All the patients were followed up for 6-18 months [(10.2±4.2)months]. The mortality rate during the acute phase (within 30 days after surgery) was 2.37% with 12 deaths due to hemorrhagic shock, 10 due to traumatic brain injury, 6 due to multiple organ dysfunction syndrome (MODS), and 2 due to pulmonary infection. The average fracture healing time averaged 3.8-18 months [(11.5±4.2)months], with 89.49% of the patients having bone union within 12 months after surgery, 8.93% having bone union within 18 months after surgery, and 1.58% undergoing reoperation. For the patients with internal fixation failure and nonunion, the average healing time was extended to (10.2±2.2)months and (13.7±3.3)months respectively. At the last follow-up, the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, HSS knee score, and AOFAS ankle-hindfoot score were 83.93%, 90.24%, 94.12%, 85.57%, 88.46%, and 92.31% respectively, with an overall excellent and good rate of 89.11%. At 6 months after surgery, the SF-36 scores of all the patients in the eight dimensions,including the physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health were (74.4±8.6)points, (44.7±14.4)points, (77.4±10.9)points, (68.4±18.2)points, (72.5±16.0)points, (76.8±8.7)points, (49.9±17.6)points, and (72.8±17.9)points, significantly improved compared with those before operation [(63.4±12.7)points, (30.9±17.4)points, (56.4±18.0)points, (55.4±24.7)points, (53.5±21.0)points, (55.8±24.3)points, (36.9±24.0)points, (58.8±21.6)points] ( P<0.01). Complications of different degrees occurred in 214 patients (16.89%), including lung infections in 118 patients (9.31%), lower extremity deep vein thrombosis in 50(3.95%), pressure injuries in 26(2.05%), internal fixation failure in 12(0.95%), and nonunion in 8(0.63%). Conclusions:The trinity strategy provides whole-process management, personalized treatment, and overall rehabilitation for multiple orthopedic trauma. It can decrease mortality, shorten fracture healing time, improve joint function and quality of life, and reduce the incidence of complications.

Objective:The incidence of prostate cancer is increasing every year,and precision diagnosis and treatment can help reduce unnecessary prostate punctures for prostate cancer patients in the gray area.This study aims to investigate the diagnostic value of 18F-prostate specific membrane antigen(PSMA)imaging combined with prostate specific antigen(PSA)-derived indicators for gray zone prostate cancer. Methods:A total of 107 patients who underwent 18F-PSMA PET/CT imaging for suspicious prostate cancer with tPSA of 4 to 10 μg/L(PSA gray zone)in a hospital were retrospectively included,and were divided into a prostate cancer group and a non-prostate cancer group based on pathological findings.Patients underwent PSA testing,18F-PSMA,and abdominal ultrasound,and age,tPSA,fPSA,f/tPSA,prostate volume,PSA density(PSAD),maximum standardized uptake value(SUVmax),and molecular imaging prostate specific membrane antigen(miPSMA)score were compared between the 2 groups.Multivariate logistic regression was used to analyze the influencing factors the diagnosis of gray zone prostate cancer.Receiver operating characteristic(ROC)curves were constructed to evaluate the efficacy of PSAD and SUVmax alone and in combination in diagnosing gray zone prostate cancer. Results:The volume of the prostate cancer group[42.00(34.00,58.00)cm3 vs 49.00(41.27,60.41)cm3]was smaller than that of the non-prostate cancer group(Z=-2.376,P=0.017),and the PSAD[(0.18±0.06)μg/(L·cm3)vs 0.15±0.05 μg/(L·cm3)]and SUVmax[18.63(8.03,28.57)vs 9.33(5.90,13.52)]were higher than those in the non-prostate cancer group(both P<0.05).The percentage of miPSMA score≥2 in the prostate cancer group was higher than that in the non-prostate cancer group(χ2=40.987,P<0.001).PSAD(OR= 22.154,95%CI 1.430 to 873.751,P=0.042)and SUVmax(OR=1.301,95%CI 1.034 to 1.678,P=0.009)were independent influential factors for the diagnosis of prostate cancer in the gray zone.The optimal cut-off values of PSAD and SUVmax were 0.22 μg/(L·cm3)and 8.02,respectively,and the AUCs for the diagnosis of prostate cancer in the gray zone alone and in combination were 0.628(95%CI 0.530 to 0.720,P<0.05)and 0.806(95%CI 0.718 to 0.876,P<0.05),0.847(95%CI 0.765 to 0.910,P<0.05),with sensitivities of 41.03%,76.92%,and 74.36%and specificities of 79.41%,89.71%,and 92.65%,respectively. Conclusion:PSAD and SUVmax are increased in patients with gray zone prostate cancer,and the combination of PSAD and SUVmax is of high value in diagnosing gray zone prostate cancer.

【Objective】 To investigate the expression of transcription factor POU domain class 2 transcription factor 2 (POU2F2) in clear cell renal cell carcinoma (ccRCC) and human renal cancer cell lines (786-O and ACHN) and its effects on the cells’ biological behaviors such as proliferation, migration and invasion in vitro. 【Methods】 The mRNA expressions of POU2F2 in ccRCC tissues, adjacent normal tissues, cell lines 786-O and ACHN were detected with real-time polymerase chain reaction (qRT-PCR). The protein expression of POU2F2 in ccRCC tissues and adjacent normal tissues were detected with immunohistochemistry. The effects of knockdown of POU2F2 on the mRNA and protein expressions of epithelial mesenchymal transformation (EMT)-related tumor markers were detected with qRT-PCR and Western blot. 【Results】 The mRNA expression of POU2F2 in ccRCC tissues was significantly higher than that in adjacent normal tissues, and was correlated with patients’ gender, WHO/ISUP nuclear grade and TNM stage. The protein expression of POU2F2 was significantly higher in ccRCC tissues than in adjacent normal tissues, and was correlated with tumor pathological grade and TNM stage. The mRNA expression of POU2F2 was significantly decreased in 786-O cells after sh-POU2F2-1013 plasmid transfection (P<0.05); the proliferation ability, clonal formation rate, migration ability and invasion ability were significantly reduced (P<0.05). Knockdown of POU2F2 down-regulated the mRNA and protein expressions of MMP2, MMP9 and Twist in 786-O cells, while up-regulated E-ca expression. 【Conclusion】 The mRNA expression of POU2F2 was significantly up-regulated in ccRCC tissues and renal cancer cells. Knockdown of POU2F2 inhibited the proliferation, migration and invasion of cells in vitro, and slowed or inhibited the occurrence and development of renal cancer.

The high-performance liquid hromatography-quadrupole-electrostatic field orbital well high-resolution mass spectrometry method was developed for the analysis of p-methoxymethamphetamine(PMMA)and its metabolites in rabbit blood.Hypersil Gold aQ(2.1 mm×100 mm,1.9 μm)was used as the chromatographic column.The mobile phase was gradient elution with 0.1%formic acid aqueous solution(A)and 0.1%formic acid acetonitrile(B)at a flow rate of 0.3 ml/min.Electrospray ionization(ESI)with positive and negative mode scanning was used to determine p-methoxymethamphetamine and its metabolites based on excimer and secondary fragment ions.PMMA and its metabolites were characterized by high-performance liquid chromatography-high resolution mass spectrometry,which lays a foundation for the study of its pharmacodynamic substance and prevention.

The methylation of N6-methyladenosine(m6A)is an important gene expression regulation mechanism in eukaryotes.It is mainly regulated by three types of regulators:writers,erasers and readers.With the development of high-throughput sequencing technology and bioinformatics,various methods have been developed to detect and analyze m6A methylation sites.A growing body of research has shown that m6A methylation plays an important role in the occurrence and development of urological tumors,including tumor proliferation,invasion and metastasis.However,the molecular mechanism and role of m6A in different types of urological tumors have not been fully elucidated.This article reviews the main regulatory mechanism of m6A methylation modification as well as the research progress,the prognostic value and the therapeutic resistance of m6A methylation modification in urological tumors.

[摘 要] 目的：探讨叉头框蛋白D3（forkhead box protein D3，FOXD3）在胃贲门腺癌（gastric cardia adenocarcinoma，GCA）中的表达及其对SGC-7901细胞生物学行为的影响。方法：从河北医科大学第四医院生物标本库中选取2014年6月至2016年12月手术切除的49例GCA组织及相应癌旁组织标本，qRT-PCR检测FOXD3在GCA组织、癌旁组织以及在5种胃癌细胞系中（BGC-823、SGC-7901、HGC-27、MGC-803及NCI-N87）的表达。向SGC-7901细胞转染pc-DNA3.1-FOXD3或pc-DNA3.1，采用细胞增殖实验、克隆形成实验、划痕愈合实验和Transwell小室侵袭实验分别检测FOXD3过表达对SGC-7901细胞增殖、克隆形成、迁移和侵袭的影响，qRT-PCR及WB法检测细胞转染前后上皮-间质转化（epithelial-mesenchymal transition，EMT）相关分子mRNA及蛋白的表达情况，流式细胞术检测转染前后细胞周期改变。结果：GCA组织中FOXD3 mRNA的表达量明显降低，其表达水平与患者临床分期和淋巴结转移密切关联；FOXD3在胃癌细胞系中的表达均低于正常细胞（均P<0.01）。FOXD3过表达能明显抑制SGC-7901细胞的增殖、克隆形成、迁移和侵袭能力（均P<0.01），提高SGC-7901细胞中E-cadherin的表达水平，减少N-cadherin、β-catenin和vimentin的表达水平（均P<0.01），使细胞周期阻滞在G0/G1期（P<0.01）。结论: FOXD3在GCA组织中的表达明显下调，其过表达可以抑制胃癌细胞的生物学行为，FOXD3可作为抑癌基因为肿瘤治疗提供新思路。

[Abstract] Objective: To investigate the expression of lncRNA linc01503 in esophageal squamous cell carcinoma (ESCC) tissues and cell lines, as well as its effect on the proliferation, invasion and epithelial-mesenchymal transition (EMT) of ESCC cells. Methods: A total of 119 pairs of tumor tissues and corresponding para-cancerous tissues of ESCC patients were obtained from the Fourth Affiliated Hospital of Hebei Medical University between Jan. 2012 and Dec. 2014. The expression of linc01503 in ESCC tissues and cell lines (Kyse150, Kyse170, Eca109, TE1 and TE13) was detected by qPCR. The ESCC cells were transfected with pGPU6-shRNA-linc01503 or treated with TGF-β. The expressions of EMT related genes before and after transfection as well as linc01503 expression before and after TGF-β treatment were detected with qPCR. MTS and Transwell assay were performed to assess the effect of linc01503 on proliferation and invasion of ESCC cells. Results: The expression of linc01503 was significantly elevated in ESCC tissues and cell lines (all P<0.05). High expression of linc01503 was correlated with lymph node metastasis, depth of infiltration, TNM stage and the survival of ESCC patients (all P<0.05). Treatment with TGF-β promoted EMT of ESCC cells and induced a significant up-regulation of linc01503 expression. Knockdown of linc01503 significantly inhibited proliferation and invasion ability of ESCC cells; Meanwhile, the low expression of linc01503 increased the expression of E-cadherin but decreased the expressions of N-cadherin and vimentin (all P<0.05). Conclusion: lncRNA linc01503, as one of the downstream effect genes of TGF- β, promotes the proliferation, invasion and EMT process of ESCC cells.