[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 effectiveness of continuous improvement projects based on FOCUS-PDCA on the pathogen sample submission rate before antimicrobial therapy for inpatients.Methods FOCUS-PDCA was used to improve the pathogen sample submission rate in Affiliated Hospital of Nantong University from Jun.to Jul.2023.The inpatients from Jan.to May 2023(before improvement)were enrolled as control group,and the inpatients from Aug.to Dec.2023(after improvement)were enrolled as improvement group.The utilization rate of antibiotics,the submission rate of pathogen samples,the submission rate of clinical microbial samples and the detection rate of multidrug-resistant organisms were compared between the 2 groups.Results The utilization rate of therapeutic antimicrobial agents(32.18％vs 32.93％,P=0.003)and antimicrobial consumption intensity(39.99 defined daily dose[DDD]/100 patient-days vs 44.19 DDD/100 patient-days)in the improvement group were significantly lower than those in the control group.The pathogen sample submission rates before antimicrobial therapy and key antimicrobial combination therapy in the improvement group were significantly higher than those in the control group(52.01％vs 23.64％,87.74％vs 77.71％;both P＜0.001).The qualified rate of microbial specimens in the improvement group was significantly higher than that in the control group(88.77％vs 80.11％,P＜0.001).The detection rates of multidrug-resistant organisms and carbapenem-resistant Klebella pneumoniae in the improvement group were significantly lower than those in the control group(40.45％vs 48.42％,29.65％vs 43.17％;both P＜0.001).Conclusion The continuous improvement projects based on FOCUS-PDCA can improve the pathogen sample submission rate,reduce the detection rate of multidrug-resistant organisms,and promote standardized hospital infection quality management.

In order to afford new antiplatelet agents with higher potency, a series of paeonol oxime derivatives (4a-4y) were designed and synthesized from paeonol.Their structures were confirmed by HRMS, 1H NMR spectra.The anti-platelet aggregation activity of the target compounds was evaluated.The results revealed that most of them had moderate to good anti-platelet aggregation activity.Among them, compound 4h and 4j were the most potent on adenosine diphosphate (ADP)-induced platelet aggregation and collagen-induced platelet aggregation. Furthermore, the target compound 4h not only showed strong antiplatelet aggregation activity, but also exhibited good water-solubility and drug-like properties, which can be used as a new antiplatelet active compound for further research.

Objective To find out the current situation and related factors of perioperative pain management for the patients with fracture among the top three hospitals in our region.Methods A general information questionnaire was made,to survey the the status and the influencing factors of top hospital nursing work of fractures patients with perioperative pain management.The researchers label the questionnaire with the method of digital label,and the data collected are kept secret,and the statistical method is used to evaluate it.Finally,the conclusion is drawn.Results The working life of the nursing staff,nursing education,nursing staff after underwent fracture perioperative pain knowledge of continuing education is the mainly factors to affect patients surfing with fracture,perioperative pain management cycle of practice the three most important factors.Conclusion Three tertiary hospitals in the area fracture nursing of pain management need to be strengthened.we strengthen fractures patients with perioperative pain management cycle practice by training of clinical nursing staff.

Objective To investigate the effect of enriched environment (EE) on behavior and expression of mitogenactivated protein kinase phosphatase-1 (MKP-1) in hippocampus of depression rats induced by chronic unpredicted mild stress (CUS) and to provide clues for the molecular mechanism of treating depression.Methods Forty Sprague-Dawley rats were randomly divided into control group,CUS group,fluoxetine group and EE group,with 10 rats in each group.The rats in CUS group,fluoxetine group and EE group were given 8 weeks of CUS,and from the fifth week,the rats in EE group and fluoxetine group were given EE and fluoxetine for 4 weeks,respectively.The changes of behavioristic of the rats in the four groups were evaluated by body mass gain,open field test,and sucrose preference.The expression of MKP-1 in hippocampus was detected by Western blot.Results There was no significant difference in body mass,distance of horizontal movement,the number of upright,the times of passing through the grid and sucrose preference index among the four groups(P ＞ 0.05).After modeling,compared with the control group,the body mass gain,distance of horizontal movement,the number of up-right,the times of passing through the grid and sucrose preference index in the CUS group,fluoxetine group and EE group were decreased significantly(P ＜ 0.05);there was no significant difference in the body mass gain,distance of horizontal movement,the number of up-right,the times of passing through the grid and sucrose preference index among the CUS group,fluoxetine group and EE group(P ＞ 0.05).After intervening by fluoxetine and EE,the body mass gain,distance of horizontal movement,the number of up-right,the times of passing through the grid and sucrose preference index in the CUS group were lower than those in the control group(P ＜0.05);but there was no significant difference in the body mass gain,distance of horizontal movement,the number of up-right,the times of passing through the grid and sucrose preference index between the control group and the fluoxetine group and EE group(P ＞ 0.05).Compared with the CUS group,the body mass gain,distance of horizontal movement,the number of up-right,the times of passing through the grid and sucrose preference index in the fluoxetine group and EE group were higher(P ＜ 0.05);there was no significant difference in the body mass gain,distance of horizontal movement,the number of up-right,the times of passing through the grid and sucrose preference index between the fluoxetine group and EE group (P ＞ 0.05).The expression of MKP-1 in hippocampus of CUS group and EE group was higher than that in the control group (P ＜0.05).There was no significant difference in the expression of MKP-1 in hippocampus between the fluoxetine group and control group(P ＞ 0.05).Compared with the CUS group,the expression of MKP-1 in hippocampus in the fluoxetine group decreased (P ＜ 0.05).There was no significant difference in the expression of MKP-1 in hippocampus between the EE group and CUS group(P ＞0.05).Compared with the fluoxetine group,the expression of MKP-1 in hippocampus in the EE group was higher(P ＜ 0.05).Conclusion EE can significantly improve depressive symptoms in rats,but it has no significant effect on MKP-1 protein expression in hippocampus,and EE may not act on depression by affecting MKP-1.

Objective: Eosinophilic esophagitis (EoE) is a chronic immune-mediated esophageal disease.The current domestic reports of EoE in children is rare.The aim of this study was to analyze the clinical features, the diagnosis and treatment advance of EoE in children by case analysis and literature review. Method: Clinical data of 22 children with EoE from January, 2011 to December, 2015 in Children′s Hospital, Zhejiang University School of Medicine were recorded, retrospective analysis was performed on clinical presentation, gastroendoscopy and histopathological examination features and the treatment. Result: (1) Clinical data: EoE can occur at any age in children (5 months to 13 years). The most common clinical manifestations of EoE are vomiting and abdominal pain, 45% (10/22) and 41%(9/22) respectively. (2) Endoscopy and pathological features of esophageal mucosa: 11 cases with coarse mucous membrane (50%), 6 cases with congestion or erosion of esophageal membrane (27%), 5 cases with longitudinal crack (23%), 3 cases with ring uplift (14%), 3 cases with granular uplift (14%), 3 cases with normal mucosa(14%). Histopathologic manifestation is eosinophil infiltration and the eosinophil counts were all more than or equal to 15/HP. (3) Laboratory results: 13 cases had increasing eosinophil counts and eosinophils proportion (62%). (4)Allergy history: among 22 cases, 7 patients had allergy history (32%). (5) Situation of treatment and remission: 16 cases had clinical remission by oral omeprazole; 2 cases had clinical remission by oral Omeprazole and Montelukast sodium; 1 case acquired remission by elimination diet; 1 case acquired remission by elimination diet and oral prednisone. 2 cases dropped out; Only 2 patients received gastroendoscopy re-examination after 3 months and revealed esophageal mucosal histologic complete recovery. Conclusion The clinical symptoms of EoE in children varies.Esophageal mucosal features of gastroendoscopy examination in children with EoE were longitudinal crack, white exudates or plaques, paper mucosa, ring uplift and granular uplift.Most patients could achieve remission by using proton-pump inhibitors, only few children needed elimination diet and change formula, or even oral glucocorticoids.

OBJECTIVETo understand the junction protein expression of gastric mucosa including occlusal proteins (occludin), closed protein-4 (claudin-4), zonula occluden-1(ZO-1), epithelial cadherin (E-cadherin), and β ring protein (β-catenin) and the clinical significance in children with Helicobacter pylori (Hp) infection.

METHODSeventy patients in whom gastric endoscopy was performed because of nausea, vomiting, abdominal pain, bloating, acid reflux, melena, and other gastrointestinal symptoms were enrolled in this study from Dec. 2010 to Apr. 2013 in our hospital. Informed consent was signed by their parents, and the study was in accordance with the principles of medical ethics. Hp positivity was confirmed if both respiratory urea test (RUT) and Hp were positive by gastric mucosal pathology. Gastric mucosal samples from 70 patients were enrolled in this study, 23 of them were Hp negative, 47 of them were Hp positive (24 cases without peptic ulcer, 23 cases with peptic ulcer). The mRNA levels and protein expression of tight junction protein of gastric mucosa were measured by RT-PCR and Western blot respectively. The location and semi quantitative content of E-cadherin and β-catenin in gastric mucosa were detected by immunohistochemical staining method.

RESULTThe mRNA level of E-cadherin, β-catenin, ZO-1 in the Hp positive group regardless of peptic ulcer was significantly lower than that in the Hp negative group. Hp positive without peptic ulcer group were 0.0008, 0.0040, 0.0014, respectively; Hp positive with peptic ulcer group were 0.0010, 0.0090, 0.0013, respectively; Hp negative group were 0.0137, 0.0423, 0.0198, respectively (F values were 36.956, 39.893, 38.962, respectively, all P<0.05). The expression of claudin-4 mRNA in Hp positive group with peptic ulcer increased significantly, the difference among Hp positive group with peptic ulcer, Hp positive group without peptic ulcer and Hp negative group was statistically significant (0.1438 vs. 0.0926 vs. 0.0789) (F value was 11.964, P<0.05), while the difference of occludin mRNA levels among the three groups was not statistically significant.Immunohistochemistry results showed that the score of E-cadherin, β-catenin positive cell in the Hp positive patients were also significantly lower than that in the Hp negative group (t values were 3.981 and 2.340, all P<0.05, respectively). Western blot results showed that the protein levels of β-catenin in Hp positive group with peptic ulcer were significantly lower than that in Hp negative group, while the protein levels of E-cadherin in Hp positive patients regardless of peptic ulcer were decreased significantly in Hp negative group.

CONCLUSIONOur results revealed that the tight junction protein E-cadherin, β-catenin, ZO-1 expression of gastric mucosa were decreased in children with Hp infection, while claudin-4 expression was increased in Hp positive patients with peptic ulcer, suggesting that damage to gastric epithelial barrier function may be the main pathogenesis of Hp associated gastric diseases in children.

Blotting, Western ; Cadherins ; metabolism ; Child ; Claudin-4 ; metabolism ; Gastric Mucosa ; metabolism ; pathology ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Immunohistochemistry ; Occludin ; metabolism ; Peptic Ulcer ; metabolism ; microbiology ; RNA, Messenger ; Tight Junction Proteins ; metabolism ; Tight Junctions ; metabolism ; Zonula Occludens-1 Protein ; metabolism ; beta Catenin ; metabolism

Objective:To explore the effect of irbesartan in the treatment of the patients with hypertension complicated with diabetes mellitus and its influence on urine trace albumin. Methods:Totally 62 cases of patients with hypertension complicated with diabetes mel-litus were divided into the observation group and the control group randomly with 31 patients in each. The patients in the control group were given the conventional blood sugar and blood pressure control, and the patients in the observation group were treated with irbesartan additionally. The treatment course was 3 months. The changes of blood pressure and urine trace albumin of the patients in the two groups before and after the treatment were compared, and the effect of blood pressure control and the adverse effects were evaluated as well. Re-sults:The total effective rate of the observation group was 96. 8%, which was much higher than that of the control group (83. 9%, P<0. 05) , and the difference had statistical significance (P<0. 05). After the treatment, the systolic blood pressure and diastolic blood pressure of the patients in the two groups were reduced significantly (P<0. 05), and the urinary albumin excretion rate (UAER) and the level of urine trace albumin (mAlb) of the patients in the two groups were obviously lowered (P<0. 05), and the decrease in the obser-vation group was more notable than that in the control group (P<0. 05). No significant difference in the incidence of adverse reactions was shown between the two groups(P>0. 05). Conclusion:The clinical effect of irbesartan in the treatment of patients with hypertension complicated with diabetes mellitus is promising with low incidence of adverse effects,which can effectively protect renal function as well.

Objective To explore the distribution of inflam m atory cells and positive expression of P-se-lectin glycoprotein ligand-1 (PSG L-1) in infant brainstem tissue from hand-foot-m outh disease related fatal brainstem encephalitis. Methods Tw enty brainstem sam ples from infants suffered from brainstem en-cephalitis w ere collected as the experim ental group. Ten brainstem sam ples from infants died of non-brain diseases and injuries w ere collected as the control group. The distribution of inflam m atory cells and the expression of PSG L-1 in the tw o groups w ere exam ined by im m unohistochem ical m ethod. The characteristics of the positive cells w ere observed. Results In brainstem tissue of the experim ental group, there w ere sleeve infiltrations of inflam m atory cells around the vessels and in the glial nodule. Microglia was the m ost and following was neutrophils around the vessels and in the glial nodule. There was a significant statistical difference am ong m icroglias, neutrophils and lym phocytes (P<0.05). There was no sleeve infiltration in the control group. PSG L-1 protein was expressed w idely in inflam m atory cells in the experim ental group, especially in the inflam m atory cells around the vessels and in the glial nodule. B ut PSG L-1 positive staining could be observed significantly less in the control group com paring with the experim ental group (P<0.05). Conclusion Microglia is the m ain type of inflam m atory cells involved in the progress of the fatal disease. Moreover, PSG L-1 could participate in the pathogenesis of hand-foot-m outh disease related fatal brainstem encephalitis.