[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。

OBJECTIVETo analyze the audiological features and genetic background of patients carrying mitochondrial DNA(mtDNA) 1555A>G mutation and factors which may influence the extent of nonsyndromic hearing loss associated with the mutation.

METHODSA literature search was carried out on databases including PubMed, CNKI, Wanfang, and VIP. Combined with author's data, the clinical features of the patients, in particular audiological characteristics, were summarized.

RESULTSA total of 857 effective cases were collected and analyzed. A significantly correlation was identified between history of aminoglycosides exposure and extent of hearing loss, in addition with a negative correlation between the age of onset and extent of hearing-impairment. Drug exposure was corelated with the age of onset but independent to the loss of high-frequency hearing loss. Heteroplasmies had a reverse correlation with the degree of hearing loss. Among the haplotypes of mitochondrial DNA, haplotype D was the most common one, while haplotype B had the highest penetrance.

CONCLUSIONNonsyndromic hearing loss associated with mitochondrial DNA 1555A>G mutation is influenced by factors such as aminoglycosides exposure, age, proportion of mutation, and haplotype of the mitochondrial DNA. Analysis of clinical cases is critical for identifying individuals carrying deafness susceptibility mutations and is the first step for early diagnosis.

Objective: To investigate the effect of the treatments for obstructive sleep apnea hypopnea syndrome (OSAHS) on the resistant hypertension (RH) of patients. Methods: Eighty patients with OSAHS and RH (blood pressure could not be controlled under 140/90 mmHg (1 mmHg=0.133 kPa) even with more than three kinds of antihypertensive drugs including diuretics) received surgery or continuous positive airway pressure (CPAP) treatment. The results of polysomnography monitoring, ambulatory blood pressure monitoring, and the dosage of antihypertensive medication were recorded before and six months after the treatment. Results: Apnea hypopnea index (AHI) decreased from (32.9±10.8) before treatment to (9.4±6.5) after treatment, while the lowest oxygen saturation (SaO₂) increased from (0.682±0.062) to (0.884±0.056), with significant differences (t value was 18.863 and 26.614, respectively; both P<0.001). Twenty-four hours systolic blood pressure (SBP)/diastolic blood pressure (DBP) decreased respectively from ((150.5±9.8)/(97.8±7.3)) mmHg to ((140.7±6.8)/(88.6±6.3)) mmHg, daytime SBP/DBP decreased from ((154.3±8.9)/(100.6±7.4)) mmHg to ((144.8±5.8)/(91.3±5.5)) mmHg, and nighttime SBP/DBP decreased from ((145.5±8.8)/(93.8±6.4)) mmHg to ((135.8±5.7)/(84.6±5.9)) mmHg, with significant differences (t value was 7.832, 6.903, 7.005, 6.848, 8.025, 7.554, respectively; all P<0.001). The reduction of nighttime SBP /DBP was ((11.5±2.2)/(10.2±3.1)) mmHg, and the reduction of daytime SBP/DBP was ((9.0±2.8)/(7.9±3.5)) mmHg. The reduction of nighttime SBP/DBP was more obvious than daytime SBP/DBP, with significant differences (t value was 9.732 and 6.936, respectively; both P<0.001). Before treatment, nighttime blood pressure decrease rate below 10% was showed in 75 percent of patients, and after treatment, this rate only in 37.5 percent of patients (χ²=22.857, P<0.01). The numbers of required antihypertensive drugs decreased in 45 (56.3%) cases, the average numbers of antihypertensive drugs decreased from (3.2±0.4) before treatment to (2.6±0.5) after treatment, with a significant difference (t=9.276, P<0.01). Conclusions After treatment of OSAHS, the blood pressure of the patients with OSAHS and RH dropped significantly, the circadian rhythm of blood pressure condition was better, the varieties of antihypertensive drugs taken in these patients were reduced significantly.

A new method for chiral separation and purity inspection of landiolol hydrochloride and its stereoisomers was developed by ultra-performance convergence chromatography ( UPC2 ) . The mobile phase was the mixture of supercritical CO2 and methanol/n-butyl alcohol/acetonitrile (1:1:1, V/V) plus 0. 5%NH3?H2O. The separation was carried out on the Daicel CHIRALPAK? IF column (150 mm × 4. 6 mm, 3 μm) with a flow rate of 2. 8 mL/min at 50℃ using 223 nm as detection wavelength. Under the optimized experimental conditions, for R,R-stereoisomer, R,S-stereoisomer and S,R-stereoisomer, the detection limits (LOD, S/N=3) were 0. 3, 0. 4 and 0. 3 mg/L, the linear ranges were 2-300 mg/L, 5-300 mg/L and 2-300 mg/L, the recoveries of spike samples were 103. 4%±2. 5%, 91. 8%±2. 5% and 101. 7%±1. 5%, and the injection repeatabilities were 0. 06%, 0. 09% and 0. 08% (n=6), respectively. The experimental results demonstrate that the UPC2-based method can be used for the analysis and determination of landiolol hydrochloride and its stereoisomers.

Objective: To evaluate the impact of intracoronary administration of eptifibatide oncoronary no-reflow and myocardium perfusion in patients with ST-elevation myocardial infarction (STEMI) at percutaneous coronary intervention (PCI). Methods: A total of 80 STEMI patients with emergent PCI were randomly divided into 2 groups: Eptifibatide group, the patients received intracoronary administration of eptiifbatide and Control group, the patients received the same volume of normal saline.n=40 in each group. The baseline condition, post-operative vascular recanalization, changes of platelet aggression at pre- and post-medication were compared between 2 groups. Echocardiography was examined at immediately and 24 weeks after operation;myocardial infusion imaging was examined at l week after operation. All patients were followed-up for 24 weeks to observe the incidence of major adverse cardiovascular events (MACE). Results: Compared with Control group, Eptifibatide group showed increased ratios of post-operative TIMI grade 3 (72.5%vs 92.5%) and myocardium perfusion (70.0% vs 90.0%), bothP<0.05; decreased post-operative and 2h post-medicinal platelet aggression and they were both lower than Control group at the same period, allP<0.05. Eptiifbatide group had obviously improved LVEDD and LVEF at 24-week than 1-week after PCI and they were both superior to Control group, allP<0.05. There were 7 (17.5%) patients in Eptiifbatide group and 7 (7.5%) in Control group suffering from small bleeding events, P>0.05; no severe bleeding eventand no in-hospital thrombocytopeniaoccurred. MACE occurrence rates during 24-week follow-up period were 12.5% vs 22.5%, P>0.05. Conclusion: Intracoronary administration of eptiifbatide in STEMI patients at emergent PCI could effectively improve coronary blood lfow,increase myocardium perfusion and enhance cardiac function without severe bleeding events.

Objective To analyse the survival time and related factors of patients with brain stem glioma who received 3DCRT.Methods Thirty-six patients with brain stem tumor were admitted from October 2004 to December 2008 and all received 3D-CRT with the dosage (50-54 Gy,25-30 f,5-6 weeks).During treatment,the patients’ outcomes were analyzed by observing the changes of symptoms,signs and adverse radiotherapy reaction and all of them were followed-up in the next 3 years.The survival data were analyzed by Kaplan-Meire method.Results The median survival time was 9 months in the 23 pediatric patients and 15 months in 13 adult patients.One-,two-and three-year survival rates between pediatric group and the adult group were 43.5 ％ (10/13) vs 76.9 ％ (10/13),26.1％ (6/23) vs 46.2 ％ (6/13),8.7 ％ (2/23) vs 38.5 ％ (5/13).Karnofsky performance scale score at admission (x2 =20.059,P =0.000),tumor site (x2 =17.585,P =0.000),growth pattern (x2 =21.247,P =0.000) were associate with survival time.Conclusion 3DCRT is an effective therapy to brain stem glioma,childhood onset,pontine glioma,diffusion style and Karnofsky performance scale less than 80 are risk factors of poor prognosis.

Epidermal growth factor receptor (EGFR) and its ligands (EGF and TGFalpha) are over-expressed in a variety of tumors. Immunization EGF-carrier protein inhibits tumor growth through abrogating binding of EGF to EGFR. Here, a chimeric protein of EGF and TGFalpha (E5T) was genetically fused to Staphylococcal enterotoxin A (SEA), a bacterial superantigenic protein which promotes humoral B cell response through enhancement of Ag-specific CD4 T cells activity. The resulted fusion proteins were expressed in Escherichia coli and purified though metal chelating affinity chromatography. Immunization of E5T-mSEA fusion protein in mice induced production of high titers antibodies, which recognize both EGF and TGFalpha. Anti- E5T-mSEA serum at dilution of 1:10 significantly inhibited growth of A431 cell lines but had little effect on 293T cell lines.
Amino Acid Sequence ; Animals ; Cancer Vaccines ; biosynthesis ; immunology ; Cell Line, Tumor ; Enterotoxins ; biosynthesis ; genetics ; Epidermal Growth Factor ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunization ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Random Allocation ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Transforming Growth Factor alpha ; biosynthesis ; genetics

Radioimmunotherapy (RIT) is one kind of targeted immunotherapy. It is a better therapy of tumour treatment for its distinct advantages. Nowadays, it has been widely used to treat lymphoma, and part of solid tumors. In this review,basic research and clinical application related to solid tumor are summarized.

Objective To evaluate biodistribution and pharmacokinetics pattern of ~(131)I-labeled rch24which is the region-grafted (humanized) anti-carcinoembryonic antigen (CEA) monoclonal antibody in nude mice. Methods Nude mice bearing cancer xenografts received intravenous injections of ~(131)I- rch24, then blood, plasma, heart, liver, spleen, lung, kidney, tumor and other tissues were taken at different time point for determination the concentration of radioactivity and calculate the T/NT value. Nude mice were packeted randomly to four group of high, medium, low dose and continuous administration, blood drug concentration was detected by ELISA method at the different intervals. Then, draw the concentration-time curve and calculate the pharmacokinetics paramete. Results After administration, radioactivity of the tumour was significantly enhanced whereas radioactivity of normal tissues decreased gradually. For single administration, at the dose of low to medium, pharmacokinetics pattern was linearity -kinetics whereas for high dose group,pharmacokinetics paramete shown some behavior of non-linearity-kinetics. Conclusion Our results suggest that the ~(131)I-labeled region-grafted (humanized) anti-CEA monoclonal antibody rch24 exhibit a considerable targeting activity so as to ~(131)I radioisotopes can be concentrated specifically in tumor. The pharmacokinetics pattern of this medicine was different at different dose.