[摘 要] 目的：探讨RNA结合蛋白15（RBM15）通过三磷酸腺苷酶家族蛋白3A（ATAD3A）调控Wnt/β-catenin通路对宫颈癌细胞恶性生物学行为的影响。方法：利用TCGA数据库分析宫颈癌组织中RBM15 mRNA表达水平及其与患者预后的关系。收集2024年1月至10月期间在赣州市人民医院手术切除的32例宫颈癌组织及癌旁组织标本，以及宫颈癌细胞HeLa、MS-751、C-33A和SiHa，通过免疫组化、WB法检测宫颈癌组织和细胞中RBM15的表达水平。利用SRAMP在线数据库筛选ATAD3A mRNA的m6A修饰位点。通过RNA免疫沉淀实验、RNA衰变实验及挽救实验鉴定RBM15与ATAD3A mRNA的相互作用。采用RNA干扰技术和病毒感染技术，在宫颈癌HeLa和SiHa细胞敲低或过表达RBM15和ATAD3A，qPCR和WB法检测mRNA和蛋白表达，CCK-8法、划痕实验和Transwell实验检测各组细胞的增殖、迁移和侵袭能力。结果：宫颈癌组织中RBM15 mRNA和蛋白阳性率均显著高于癌旁组织（均P < 0.001）。宫颈癌HeLa、MS-751、C-33A和SiHa细胞RBM15蛋白表达水平显著高于正常宫颈细胞Ect1/E6E7（均P < 0.01）。RBM15 mRNA高表达组患者5年无进展生存率低于低表达组患者（P < 0.01）。宫颈癌组织中ATAD3A的表达水平显著高于癌旁组织（P < 0.001），RBM15 mRNA与ATAD3A mRNA呈正相关（r = 0.601，P < 0.05）。ATAD3A mRNA的501、5 312、12 137位点存在高可信度的m6A修饰位点。HeLa、SiHa细胞中过表达RBM15后，ATAD3A mRNA和蛋白表达升高，而敲低RBM15后，ATAD3A mRNA和蛋白表达降低（均P < 0.001）。RNA免疫沉淀实验显示，与IgG组相比，RBM15抗体的免疫沉淀中ATAD3A mRNA显著富集（均P < 0.001）。MeRIP-qPCR实验显示，ATAD3A mRNA 501、5 312、12 137位点均存在明显的m6A甲基化富集（均P < 0.001）。RNA衰变实验显示，敲低RBM15能降低HeLa、SiHa细胞ATAD3A mRNA的半衰期和稳定性（均P < 0.001）。敲低HeLa、SiHa细胞RBM15的表达，显著抑制癌细胞的增殖、迁移及侵袭，Wnt/β-catenin通路相关蛋白Wnt3、β-catenin、vimentin表达均显著降低（均P < 0.001）；而过表达ATAD3A可完全逆转上述抑制作用（均P < 0.001）。结论：RBM15通过m6A修饰ATAD3A mRNA调控Wnt/β-catenin信号通路，从而促进宫颈癌细胞增殖、迁移及侵袭。

Objective:To investigate the common disease spectrum of grass-root troops, to provide a basis for meeting the needs for military medical support among officers and soldiers, and to improve targeted training of military general practitioners in grass-root troops.Methods:With reference to International Classification of Diseases (9th edition), a questionnaire survey was conducted to investigate 146 types of diseases among the military doctors in grass-root troops. SPSS 23.0 was used to determine the score of the common degree of each disease, and the mean score of each disease was ranked. At the same time, the Kendell consistency test was used to compare the scores of doctors between different altitudes and troops.Results:Most grass-root military doctors were male graduates aged 20-39 years, among whom 109 (51.90%) did not receive continuing education, and there were 140 doctors (66.67%) belonging to the health company, 11 (5.24%) belonging to the health platoon, 35 (16.67%) belonging to the health clinic, and 24 (11.42%) belonging to the health team. As for the location of the troops, there were 42 doctors (20.00%) in the temperate zone, 56 (26.67%) in the subtropical zone, 57 (27.14%) in the alpine region, 37 (17.62%) in the plains, and 18 (8.57%) on the islands. There were 173 patients with an age of 20-29 years, accounting for 82.38%. The Kendell consistency test showed Kendell's W=0.968 ( χ 2=711.51, P<0.001), suggesting a consistent evaluation of 146 diseases by grass-roots doctors from different altitudes. Conclusion:With reference to the investigation of disease spectrum, it is necessary to formulate characteristic training objectives and contents for the training of military general practitioners, explore the training models and methods for military general practitioners in grass-root troops, promote the construction of military general practitioners during the new military reform, and improve the post competency of military doctors.

Abdomen/diagnostic imaging* ; Artificial Intelligence ; Body Composition ; Magnetic Resonance Imaging ; Practice Guidelines as Topic

Objective:To explore solutions to the "grey zone" of activated partial thromboplastin time (APTT) mixing study, and establish the clinical application pathway of it.Methods:Patients treated in West China Hospital of Sichuan University from January 1, 2018, to December 31, 2019, with a prolonged APTT were included in this study. The ROC curve was used to analyze the"cut-off"of different methods and explore solutions to the "grey zone" by combination of the 1∶1 and 4∶1 mixing study. Similar samples from January 1, 2020 to December 31, 2020 were included to verify the diagnostic efficiency of the clinical application pathway.Results:The traditional Rosner index criterion had a low diagnostic accuracy in differentiating factor deficiencies from inhibitors. A total of 49 cases (15%) in the establishment group and validation group were located in the "grey zone". The optimal cut-off value of the Rosner index in our 1∶1 mixing study for determining factor deficiency was 5.0%, and inhibitor was 9.1%. The sample between 5.0% and 9.0% needed 4∶1 mixing studies, which could significantly improve the detection sensitivity of inhibitors. The percentage of extended time after incubation-P (1∶1 mixing>10.8% and 4∶1 mixing>13.5%) was better than the traditional criterion mentioned by"consensus"in determining whether the inhibitor was time-dependent. The sensitivity, specificity, positive predictive value and negative predictive value of combined the 1∶1 and 4∶1 mixing study in differentiating factor deficiencies from inhibitors all attained more than 90%. Only 7% (3/43)of inhibitors were incorrectly classified into the factor deficiency group by the combination, which was 20.9% (9/43) by traditional criterion. The specificity for detecting time-dependent inhibitor was increased from 54.2% to 100%, and accuracy was increased from 63.3% to 97.4%.Conclusions:The combination of 1∶1 and 4∶1 mixing study can better resolve the "grey zone". The established clinical application pathway is beneficial for the further promotion and clinical application of APTT mixing study.

Objectives To design an objective and structured evaluation system for the clinical competence of orthopedic postgraduates in the diagnosis and treatment of distal radius fractures, and to ana-lyze its reliability and validity. Methods 28 orthopaedic postgraduates representing six levels of surgical training were tested for competence in performing surgical approach for distal radius fracture on cadaver specimens during which four measures were used to assess competency: examination of basic theory based on network item bank, objective structured operation assessment,overall assessment and operation examina-tion results. In addition, the time for completion of the surgery was also recorded. Each assessment tool was correlated with the others as well as with the resident’s level of training. Results There was a significant correlation between the seniority of candidates and the score of theoretical examination (F=6.193, P=0.000), the score of structured operation examination (F=6.374, P=0.002), the score of overall assessment (F=2.321, P=0.030), and the passing rate of final operation examination (F=36.300, P=0.000). No significant differ- ences were found between seniority and time to completion of the surgical approach exposure (F=2.282, P<0.073). Conclusions The results of the present study suggested that both theoretical examination and cadaver testing discriminate between novice and accomplished postgraduates. However, although the theo-retical test scores could predict the operational test results, but the theoretical results can not guarantee excellent operational skills.

Objective To explore the application of mixed anesthesia with epinephrine hydrochloride and lidocaine in se-nile ptosis surgery.Methods 90 senile ptosis cases(180 eyes)treated from January 2016 to June 2016 were randomly divided into experimental group and control group.Subcutaneous infiltration anesthesia with lidocaine and epinephrine hydrochloride was used in the experimental group,while the control group received lidocaine only.The heart rate,blood pressure,blood loss,visual field definition,operation time,incision state,cornea and recovery rate of the two groups were observed.Results① The systolic blood pressure of the patients in the experimental group was significantly higher than that of the control group (P<0.05).There was no significant difference between two groups in diastolic blood pressure and heart rate(P>0.05).②The experimental group achieved better results than the control group in operation time,anesthesia time,amount of bleeding, visual field clarity,eyelid congestion and wound healing(P<0.05);③ There was no significant difference(P> 0.05)be-tween two groups in incisional dressing bleeding,fluid seepage and cornea status.Conclusion The mixed anesthesia of epi-nephrine hydrochloride and lidocaine had little effect on the patients' diastolic blood pressure and heart rate.It reduced the blood loss during operation and the postoperative eyelid congestion.The clarity of intraoperative vision and recovery time were also improved with the mixed anesthesia.

Objective: To evaluate the differential calretinin immunostaining in different segments of total colonic aganglionosis and its utility in the diagnosis. Methods: Nine specimens including ileum and colon segments were obtained from 9 patients with total colonic aganglionosis (TCA), from 2010 to 2016 year, in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology. Another 9 ganglionic specimens including the same segments from patients with non-Hirschsprung disease (non-HD) patients were collected as control. All cases were immunostained with calretinin. The patterns of calretinin immunostaining were observed, and morphometric analysis of each sample was performed by image analysis program (Image-Pro-Plus). The mean absorbance was evaluated by calculating the areas of the lamina propria occupied by the positively stained area of the calretinin at high power field. Results: The same pattern of calretinin immunostaining was seen in ganglionic ileum and ganglionic colon segments, with staining seen in intrinsic nerves fibers (INF), and in granular aggregates in the lamina propria and muscularis mucosae. There was no significant difference in the numbers of calretinin-positive INF from the ganglionic segments. In contrast, the number of calretinin-positive INF and granular aggregates in aganglionic segments were significantly lower than those in the ganglionic group (P<0.01). In the ileum transitional zone, scattered calretinin staining was observed, and the amount of calretinin-positive INF was significantly lower than those in the proximal segment of ganlionic ileum (P<0.01). Conclusions Since there is significant different expression of calretinin among the different segments from TCA, calretinin immunostaining has potential value in detecting TCA. It could be an important adjunctive method in detecting TCA in the future.

Objective: To assess the status of handgrip strength of elderly population from longevity areas in China, and to analyze the correlative factors of handgrip strength of elderly people. Methods: Data from Chinese Longitudinal Healthy Longevity Survey (CLHLS) in 2012 was used, from which1 967 participants aged ≥60 years old with valid data of grip strength value from 8 Chinese longevity areas were included. Information on demographics characteristic, life style and health status was collected using questionnaires. The handgrip strength of both left and right hands were measured by grip dynamometer. The different characteristics of group of participants with different grip strength were compared and then analyzed by adopting the Cumulative odds Logistic regression model to identify main factors associated with hand grip strength. Results: The P₅₀ (P₂₅-P₅₀) of hand grip strength of elderly people from the eight longevity areas was 20 (11-28) kg; The hand grip strength of males was 26 (18-34) kg, which was higher than that of females(14 (9-20) kg) (P<0.001). Cumulative odds Logistic regression model showed that the hand grip strength of females was lower than males, whose β value (95%CI) was-1.22 (-1.43--1.00). The elderly who was at a higher age, smoking, drinking or with anemia, had a comparatively lower handgrip strength, whose β (95%CI) value were separately-0.08(-0.09~-0.07),-0.29(-0.56~-0.02),-0.54(-0.80~-0.28), and-0.41(-0.62~-0.20). And the elderly who had a higher boby mass index, drinking tea and outdoor activities, had a comparatively higher handgrip strength, whose β(95%CI) value were separately 0.28 (0.15-0.40), 0.25(0.03-0.47) and 0.51(0.30-0.71). Conclusion Age and gender were the main correlative factors, lifestyles and physical conditions might also be correlative factors of hand grip strength of the elderly from longevity areas in China.

Objective: To explore the association between biomarkers and activities of daily living (ADL) in the elderly over 65 years old from longevity areas in China. Methods: A total of 2 439 people from 8 longevity areas were included in our baseline survey in 2012. Using questionnaires, body measurements, and blood biochemical examinations, information on demographics characteristic, life style, ADL, blood pressure and biomarkers were collected. Based on these six items of ADL (bathing, dressing, indoor activities, toileting, eating, bowel and bladder control), we constructed a dichotomous indicator for ADL. A respondent was defined as ADL disabled if any difficulty in one or more of the above six activities was reported. Information were collected in the follow-up in 2014 using the same questionnaires and examinations. We excluded information on the elderly who lacked ADL or biomarkers test results or with ADL disability at baseline study. Finally 938 elderly people over 65 years old were included in this analysis. Multivariate logistic regression model was used to analyze the influence factors of ADL disability. Results: During the 2-year follow-up, 100 (10.7%) participants developed into ADL disability, with a rate at 10.7%. Multivariate logistic regression analysis indicated that each year increase in age or each 1 mmHg (1 mmHg=0.133 kPa) increase in systolic blood pressure (SBP) would cause the risk of ADL disability to increase 9% or 1%, whose OR (95%CI) were separately 1.09 (1.06-1.12), 1.01 (1.00-1.02). Han nationality or cognitive impairment increased the risk of ADL disability, whose OR (95%CI) values were separately 4.90 (1.13-21.24), 2.47 (1.44-4.25), while increased lymphocyte count (>1.60×10⁹/L), being married, or participating in recreational activities decreased the risk of ADL disability, whose OR (95%CI) values were separately 0.51 (0.31-0.82), 0.52 (0.28-0.96), 0.43 (0.23-0.80). Conclusion In the elderly elevated lymphocyte count was associated with lower risk of ADL disability. In addition, incresed age, increased SBP, unmarried, Han nationality or cognitive impairment were associated with the increasing risk of ADL disability in older people, while participating in recreational activities would reduce the risk.

Objective: To investigate the influence factors of survival outcome among elderly aged ≥80 years old. Methods: In baseline survey in 2009, 930 participants aged ≥80 years old were enrolled from 7 longevity areas, to collect the information of socioeconomic factors, life style, cognitive function, activities of daily living and diseases, as well as physical examination to test biomarkers of blood and urine. The survival status was followed up at 2012 and 2014 survey. Stepwise Cox proportional hazards models were used to screen influence factors of 5-year survival. Results: During 5 years of follow-up, 571 participants died, 133 participants were lost to follow up, and the all-cause mortality was 63.4%. In stepwise Cox proportional hazards models, male, unmarried, self-reported poor life quality, disability in daily life, cognitive impairment, cardiovascular and cerebrovascular diseases, chronic kidney diseases were risk factors for elderly survival outcome, with the HR (95%CI) at 1.75 (1.40-2.12), 1.49 (1.10-2.03), 1.40 (1.16-1.69), 1.37 (1.11-1.70), 1.51 (1.22-1.88), 1.62 (1.18-2.23) and 1.48 (1.23-1.77) respectively. Each 1 year increase in age corresponded to 4% increase in mortality risk (HR (95%CI)=1.04 (1.02-1.05)); each 1 kg/m² increase in BMI corresponded to 5% increase in mortality risk (HR (95%CI)=0.95 (0.93-0.98)); each 1.0×10⁹/L increase in total lymphocyte count (TLC) corresponded to 13% increase in mortality risk (HR (95%CI)=0.87 (0.76-0.99)). Additionally, the mortality risk decreased 19% (HR (95%CI)=0.81 (0.66-0.98)) in participants with regularly physical exercise compared to those without; and the mortality risk decreased 41% (HR (95% CI)=0.59 (0.40-0.88)) in participants with elevated triglycerides (TG, ≥2.26 mmol/L) compared to those without. Conclusion In Chinese longevity areas, some nutritional and immune indices such as relatively higher level of BMI, TLC and TG were independent protective factors for 5-year survival outcome, which was different from general adults and younger elderly.