[摘 要] 目的：探讨RNA结合基序蛋白15（RBM15）是否通过含跨膜Bax抑制基序蛋白6（TMBIM6）/TGF-β/Smad轴促进宫颈癌细胞增殖、迁移及侵袭。方法：收集2023年1月至2025年1月在苏州大学附属张家港医院接受手术治疗的82例宫颈癌患者的肿瘤组织与癌旁组织。应用R语言分析癌症基因组图谱（TCGA）数据库中RBM15、TMBIM6 mRNA的表达；采用免疫组织化学法和Western blotting法检测RBM15、TMBIM6蛋白表达；采用Kaplan-Meier曲线分析RBM15、TMBIM6 mRNA表达与宫颈癌患者预后的关系；采用CCK-8法、划痕实验及Transwell侵袭实验检测RBM15、TMBIM6对宫颈癌细胞增殖、迁移及侵袭的影响；利用基于序列的RNA腺苷甲基化位点预测工具（SRAMP）预测TMBIM6 mRNA的N6-甲基腺苷（m6A）修饰位点；采用RNA免疫共沉淀（RIP）实验、RNA衰变和挽救实验鉴定RBM15与TMBIM6 mRNA的相互作用；采用甲基化RNA免疫共沉淀定量PCR（MeRIP-qPCR）检测TMBIM6 mRNA的m6A富集；采用Western blotting法检测RBM15通过TMBIM6对宫颈癌细胞TGF-β/Smad通路相关蛋白表达的影响。结果：宫颈癌组织中RBM15 mRNA、TMBIM6 mRNA表达均高于癌旁组织（均P < 0.05），癌组织RBM15、TMBIM6阳性率均高于癌旁组织（均P < 0.001）。宫颈癌细胞系HeLa、MS-751、SiHa中RBM15、TMBIM6蛋白表达均高于正常宫颈上皮细胞系Ect1/E6E7（均P < 0.05）。国际妇产科联盟（FIGO）分期ⅠB2～ⅡB期宫颈癌组织中RBM15、TMBIM6蛋白阳性率高于ⅠA～ⅠB1期（P均 < 0.05）。RBM15 mRNA高表达组、TMBIM6 mRNA高表达组5年无进展生存率分别低于各自的低表达组（均P < 0.05）。宫颈癌组织中RBM15 mRNA与TMBIM6 mRNA表达呈正相关（P < 0.05）。SRAMP预测结果显示，TMBIM6 mRNA序列的960、24876、25203位点为高可信度m6A修饰位点。在HeLa、SiHa细胞中过表达RBM15后，TMBIM6 mRNA和蛋白表达升高（P < 0.05）；敲低RBM15后，TMBIM6 mRNA和蛋白表达降低（P < 0.05）。与IgG组相比，RBM15抗体免疫沉淀物中TMBIM6 mRNA显著富集。MeRIP-qPCR结果显示，TMBIM6 mRNA的960、24876、25203位点均存在明显的m6A富集。敲低RBM15可缩短TMBIM6 mRNA半衰期并降低其稳定性。敲低RBM15可抑制宫颈癌细胞增殖、迁移及侵袭，过表达TMBIM6可逆转上述影响。在HeLa、SiHa细胞中过表达TMBIM6后，TGF-β1、Smad2、Smad3蛋白表达升高；敲低RBM15后，上述蛋白表达降低，过表达TMBIM6可逆转该影响。结论：RBM15可能通过m6A修饰提高TMBIM6 mRNA稳定性并激活TGF-β/Smad通路，从而促进宫颈癌细胞增殖、迁移及侵袭。

Objective:To investigate the promoting effect of individualized instruction combined with a blended learning model (IIBLM) in continuing training of neurology.Methods:A total of 93 trainees who received continuing training in Department of Neurology, The First Affiliated Hospital of Sun Yat-sen University, from August 2022 to December 2024 were enrolled as subjects. The 50 trainees registered since January 2024 were enrolled as observation group and received IIBLM teaching, including sub-specialty modular training, a cycle-adaptive cultivation system, a "mutual-selection" mentorship program, an on/off-line dual-track curriculum system, a dynamic course allocation mechanism based on "mutual selection", and a competency growth evaluation system, while the 43 trainees registered before January 2024 were enrolled as control group and received traditional teaching. A questionnaire survey and comprehensive competency assessments were performed to evaluate the effect of teaching, and the t-test, the chi-square test, and the qualitative analysis were used for statistical analysis. Results:Compared with the control group, the experimental group showed systematic improvements in clinical contents, theoretical curriculum, faculty competency, and workflow arrangement during continuing training, with a significant difference in the score of workflow arrangement between the two groups [(9.58±0.67) vs. (9.07±1.44), t=-2.13, P=0.037]. The observation group had a score of (97.70±1.30) for dynamic course allocation, an overall satisfaction rate of 97.15%, and a course benefit rate of 97.55%. The qualitative analysis showed that the trainees in the control group mainly complained of course monotony, while those in the observation group expected to enhance interdisciplinary integration and the cultivation of scientific research abilities. In addition, the results of competency assessment showed a continuous improvement in teaching effect after reform, with no significant difference. Conclusions:IIBLM teaching effectively enhances professional qualities, clinical competency, and the degree of satisfaction with courses among the trainees receiving continuing training, and it also revealed the necessity of interdisciplinary collaborative teaching and the integration of research and clinical practice.

Objective To investigate the guiding value and safety of three tracers in identifying sentinel lymph nodes(SLN)for lymph node dissection during radical surgery for early-stage cervical cancer.Methods A total of 114 patients with early-stage cervical cancer from January 2021 to No-vember 2022 were selected as the study subjects.They were randomly divided into methylene blue group,nano-carbon group,and indocyanine green(ICG)group according to the random number ta-ble method,with 38 cases in each group.All three groups underwent radical surgery.The methylene blue group used methylene blue for SLN identification during surgery,the nano-carbon group used nano-carbon,and the ICG group used ICG fluorescence imaging.The SLN identification status and surgery-related indicators were compared among the three groups.Trauma indicators[serum cortisol(Cor),epinephrine(E),adrenocorticotropic hormone(ACTH),and C-reactive protein(CRP)]were compared among the three groups before surgery and on 1,3,and 7 d after surgery.Tumor markers[carcinoembryonic antigen(CEA),alpha-fetoprotein(AFP),carbohydrate antigen 125(CA125),and carbohydrate antigen 199(CA199)]were compared among the three groups before surgery and 3 months after surgery.The complication rates and recurrence rates were also compared among the three groups.Results The tracing time in the ICG group was shorter than that in the nano-carbon and methylene blue groups,and the tracing time in the nano-carbon group was shorter than that in the methylene blue group(P＜0.05).In terms of the success rate and number of identified SLN,the nano-carbon group was higher than the ICG and methylene blue groups,and the ICG group was higher than the methylene blue group(P＜0.05).Regarding the SLN resection time,the nano-car-bon group had the shortest time,and the methylene blue group had the longest time,with a statisti-cally significant difference(P＜0.05).The serum levels of CEA,AFP,CA125,and CA199 in the three groups decreased significantly 3 months after surgery compared with those before surgery(P＜0.05).The within-group effects of one-way repeated-measures analysis of variance showed that the serum levels of Cor,E,ACTH,and CRP in the three groups increased first and then decreased on 1,3,and 7 d,with statistically significant differences(P＜0.05).There was no statistically signif-icant difference in the complication rates among the three groups(P＞0.05).During the 1-year fol-low-up,2 patients were lost during follow-up in the nano-carbon group,1 in the ICG group,and 2 in the methylene blue group.No recurrence was found in any of the three groups.Conclusion Among the three tracers,ICG has the shortest time for tracing SLN during radical surgery for early-stage cer-vical cancer.Nano-carbon exhibits relatively high application value compared with the other two tracers,and it guides the scope of lymph node dissection and demonstrates certain safety.

Objective:To investigate the promoting effect of individualized instruction combined with a blended learning model (IIBLM) in continuing training of neurology.Methods:A total of 93 trainees who received continuing training in Department of Neurology, The First Affiliated Hospital of Sun Yat-sen University, from August 2022 to December 2024 were enrolled as subjects. The 50 trainees registered since January 2024 were enrolled as observation group and received IIBLM teaching, including sub-specialty modular training, a cycle-adaptive cultivation system, a "mutual-selection" mentorship program, an on/off-line dual-track curriculum system, a dynamic course allocation mechanism based on "mutual selection", and a competency growth evaluation system, while the 43 trainees registered before January 2024 were enrolled as control group and received traditional teaching. A questionnaire survey and comprehensive competency assessments were performed to evaluate the effect of teaching, and the t-test, the chi-square test, and the qualitative analysis were used for statistical analysis. Results:Compared with the control group, the experimental group showed systematic improvements in clinical contents, theoretical curriculum, faculty competency, and workflow arrangement during continuing training, with a significant difference in the score of workflow arrangement between the two groups [(9.58±0.67) vs. (9.07±1.44), t=-2.13, P=0.037]. The observation group had a score of (97.70±1.30) for dynamic course allocation, an overall satisfaction rate of 97.15%, and a course benefit rate of 97.55%. The qualitative analysis showed that the trainees in the control group mainly complained of course monotony, while those in the observation group expected to enhance interdisciplinary integration and the cultivation of scientific research abilities. In addition, the results of competency assessment showed a continuous improvement in teaching effect after reform, with no significant difference. Conclusions:IIBLM teaching effectively enhances professional qualities, clinical competency, and the degree of satisfaction with courses among the trainees receiving continuing training, and it also revealed the necessity of interdisciplinary collaborative teaching and the integration of research and clinical practice.

Objective:To established a rapid method for the determination of lactose(C12 H22 O11)content.Methods:The specific rotation([α]20D)of pure lactose was measured with lactose extract product,and the correc-tion coefficient was calculated.The measured rotatory value of the sample was multiplied by the correction coeffi-cient to obtain the mass(g)of lactose in the test product,and the lactose content was calculated.The equivalence of the newly established polarimetric method and the legal test method(HPLC-RID method)for the determination of lactose content was studied.Results:The linear relation of established method was excellent with the range of 0.05-0.20 g·mL-1(r=1.000 0)and the method also had good reproducibility(RSD=0.07％(n=6)).The lactose content of 141 batches samples measured by polarimetry was compared with the results determined by official analytical procedure,and the results showed the relative deviation between the same batches was less than 1.0％.The results of one-way ANOVA also showed that there was no significant difference between two groups(Sig.＞0.05).Conclusion:The performance of polarimetry is comparable to HPLC-RID in the Chinese Pharma-copoeia 2020 Vol Ⅳ.Meanwhile,the polarimetry can reduce the test cost,shorten the test time and meet the requirements of the quality control.

Objective To explore the gene expression profile in paraspinal muscle degeneration(PMD)and identify key ferroptosis genes.Methods RNA sequencing was performed on paraspinal muscle tissue of 3 normal and 3 PMD patients respectively to obtain differentially expressed genes.Through protein-protein interaction(PPI)and gene functional enrichment analysis,the intersection of ferroptosis genes was identified to identify key hub genes associated with ferroptosis.The diagnostic value for PMD disease was analyzed by receiver operating characteristic(ROC)curves.Results A total of 292 differentially expressed genes were identified in PMD.Among them,125 genes were significantly downregulated and 167 genes were significantly upregulated.Bioinformatics analysis revealed that 14 differentially expressed genes were associated with ferroptosis.Among them,ferroptosis genes MUC1,ATF3 and CDKN1A were key hub genes with good specificity and sensitivity for diagnosing PMD.Functional enrichment analysis revealed that they may mediate the occurrence and progression of PMD by regulating cell apoptosis,ferroptosis and skeletal muscle tissue development and differentiation.Conclusion Ferroptosis genes MUC1,ATF3 and CDKN1A can serve as biomarkers for diagnosing PMD,providing theoretical basis for decoding the pathological mechanism of PMD and developing new drugs.

Dizziness or vertigo is a common clinical symptom, and its underlying etiology is complex. Many clinicians are confused about its diagnosis and treatment. This article presents a case about chronic vestibular syndrome. And case appreciation and academic discussion are conducted by well-known domestic neurologists and otologists, so as to provide a good thinking model and basic ideas for the diagnosis and treatment of dizziness or vertigo, hoping to further improve the diagnosis and treatment level among clinicians.
Humans ; Dizziness/therapy* ; Vertigo/etiology* ; Vestibular Diseases/complications* ; Otolaryngologists

Objective:To conduct a retrospective analysis of efficacy and safety of different conversion schemes of tacrolimus to slow-release dosage forms for recipients in stable phase after renal transplantation to provide rationales for the conversion strategy of tacrolimus.Methods:From January 2020 to June 2020, clinical data were reviewed for 101 kidney transplant recipients converting from common tacrolimus dosage form to tacrolimus sustained-release dosage form during postoperative stable period.There were 62 males and 49 females with an age range of 19 to 69 years.They were divided into two groups according to iso-dose and incremental-dose switching schemes.The common dosage form of tacrolimus was converted into a sustained-release dosage form with different conversion doses, They were divided into two groups of 1∶1 conversion( n=55)and >1∶1 conversion( n=46). The clinical parameters of serum creatinine(Scr), blood urea nitrogen(BUN), alanine aminotransferase(ALT)and aspartate aminotransferase(AST), alkaline phosphatase(ALP), serum albumin(ALB), white blood cell count(WBC), urinary white blood cell(UWBC), hemoglobin(Hb)and fasting blood glucose(Glu)were compared between two groups after conversion. Results:Regarding numerical change trend after switching to tacrolimus sustained-release dosage form, drug dose/variation trend was smaller and blood drug concentration more stabilized.In two subgroups converted by 1∶1 and 1>1 initial dose, change trend of dose/blood concentration in 1∶1 conversion group appeared to be more stable.However, no inter-group difference existed in long-term parameters.Scr was lower at 1 week and 3 months after switching to extended-release dosage form( P<0.05)and BUN was lower at 2 weeks( P<0.05). In addition, at 5 months after conversion, ALT and AST significantly improved as compared with common dosage form( P<0.05). Significant differences existed in urinary WBC(UWBC)at 2/3 weeks( P<0.05). After switching for 2 weeks, hemoglobin significantly improved compared with common dosage form( P<0.05). No significant differences existed in ALP, ALB or Glu at other timepoints and pre-conversion( P>0.05). In 1∶1 switch group, renal function tended to improve.At 2 weeks, BUN was lower than pre-conversion; at 1/3 weeks, Scr was lower than pre-conversion( P<0.05). In addition, there was also a trend of improvement in liver function in 1∶1 conversion group.At 1 week and 5 months, ALT was lower than pre-conversion( P<0.05). However, no significant differences existed in AST, ALB, ALP, Glu, UWBC and serum WBC count at each timepoint between two different dose conversion groups( P>0.05). After conversion, intra-individual variability of tacrolimus trough concentration significantly improved( P<0.05). Conclusions:With the same safety and efficacy as common dosage form, sustained-release dosage form of tacrolimus may improve drug variability of individuals.When converting common dosage form into sustained-release dosage form, individual differences should be considered.While monitoring trough concentrations, proper doses should be adjusted on the basis of various clinical parameters.

Objective:To explore the long-term preservation value and repair effect of normothermic machine perfusion (NMP) on clinically discarded kidneys.Methods:A case of clinical discarded donor kidney was collected, and NMP was carried out in vitro for 9 hours with recovered blood. The dynamic changes of renal appearance, blood gas and biochemistry analysis of perfusate and renal pathology were recorded. Results:In the second to fifth hour of NMP, the appearance of renal was pink and ex vivo normothermic perfusion assessment score (EVNP) was grade Ⅰ. While, the sixth hour and beyond of NMP, the appearance of kidney turned to dark red and EVNP was grade Ⅲ. The renal perfusion blood flow maintained above 150 ml/min in the first 6 hours and decreased significantly after that, and at the end, was only 50 ml/min. During the whole process of perfusion, urine output was maintained at about 100 ml/h. PO 2 remained above 100 mmHg in the first 5 hours of perfusion and from the 6th hour, was lower than 80 mmHg and continued to decline, and was close to 0 at the end of perfusion. The results showed that although the K + concentration changes in blood and urine in the first 5 hours of NMP had a good consistency, the lactic acid level had been rising. In addition, there was no significant change in the histopathology at the fourth hour of perfusion compared with that before zero-point puncture, and the fibrinous thrombus in glomeruli was improved compared with that before perfusion. However, at the sixth hour after perfusion and before the end of perfusion, the pathological changes of renal tissue were significantly worse. There were a large of thrombosis in glomerular blood vessels, renal tubular atrophy and acute tubular necrosis. Conclusions:NMP can realize the evaluation of extended criteria donors before transplantation, and it proves the feasibility and repair potential of NMP in kidney to a certain extent. At the same time, NMP also provides a new way to expand the source of donor kidney and to pre-treat organ in vitro.

Objective:To explore the effects of conditioned medium of human bone marrow mesenchymal stem cells (BMSCs) on the proliferation, adhesion and differentiation of immortalized human Müller cell line (MIO-M1).Methods:The differentiation was induced in the third-passage BMSCs with osteogenic, chondrogenic and adipogenic medium and identified by alizarin red, alcian blue and oil red O staining, respectively.The expression levels of mesenchymal stem cell markers CD73, CD90 and CD105 and hematopoietic cell markers CD34, CD45 and human leukocyte antigen-DR (HLA-DR) were assayed by flow cytometry.The expressions levels of Müller cell markers SOX9, glutamine synthetase (GS), vimentin and cellular retinaldehyde-binding protein (CRALBP), retinal stem cell markers SOX2, nestin and CHX10, and cell proliferation marker cyclin D3 (CCND3) in MIO-M1 cells were detected by immunofluorescence staining.The MIO-M1 cells were divided into standard medium group, 293T conditioned medium group, and BMSC conditioned medium group and were incubated in the medium according to grouping.The cellular area, circularity, elongation factor and perimeter were analyzed quantitatively.The cell cycle was detected by flow cytometry, and the cell proliferation was determined by neurospora experiment and 5-ethynyl-2'-deoxyuridine (EdU) staining.The expression of vascular cell adhesion molecule 1 (VCAM-1) at protein and mRNA levels in the culture supernatant was detected by enzyme linked immunosorbent assay (ELISA) and quantitative real-time PCR (qRT-PCR), respectively.The expression of retinal neuron markers protein kinase C (PKCα), Rhodopsin, microtubule-associated protein 2 (MAP2) and β-tubulin (Tuj1) was detected by immunofluorescence staining and qRT-PCR.Results:CD73, CD90, CD105 showed an enhanced expression, and CD34, CD45 and HLA-DR showed weakened expression in the BMSCs.The BMSCs differentiated into osteoblasts, chondrocytes and adipocytes.Expression of SOX9, GS, vimentin and CRALBP, SOX2, CHX10, nestin and CCND3 was found in the MIO-M1 cells.Compared with standard medium group and 293T conditioned medium group, MIO-M1 cells cultured in BMSC conditioned medium group changed into an elongated spindle-shaped or multipolar morphology with reduced cell area, increased elongation index and decreased circularity, showing statistically significant differences among them ( F=6.973, 12.370, 6.311; all at P<0.01). There were increased neurospheres formed by MIO-M1 cells in BMSC conditioned medium group compared with standard medium group and 293T conditioned medium group at different time points ( Fgroup=134.300, P<0.001; Ftime=82.910, P<0.001). Compared with the standard medium group and 293T conditioned medium group, the EdU-positive rate and proliferation index of MIO-M1 cells in BMSC conditioned medium group were significantly increased, with statistically significant differences ( F=6.973, 74.110; all at P<0.05); the VCAM-1 protein expression in cell supernatant and the relative expression level of VCAM-1 mRNA in BMSC conditioed medium group were significantly increased ( F=13.720, 7.896; all at P<0.05); the mRNA expression levels of PKCα, Rhodopsin, Tuj1 and MAP2 were higher in MIO-M1 cells of BMSC conditioned medium group under the condition of differentiation ( F=14.490, 5.424, 14.330, 7.405; all at P<0.05). Conclusions:BMSCs conditioned medium can change the morphology of MIO-M1 cells and promote their proliferation, adhesion and differentiation into retinal neurons.