Objective:To detect the changes in the levels of miR-21-5p and miR-125a-5p modified with 3′-terminal 2′-O-methylation (3′t-2′Ome) in serum extracellular vesicles (EV) of non-small cell lung cancer (NSCLC) patients, and evaluate their value as auxiliary screening molecular markers for NSCLC patients.Method:A retrospective analysis was conducted on the data of 69 NSCLC patients diagnosed at the Eastern Theater Command General Hospital from May 1st to October 31st,2023, as well as 65 age and gender matched healthy controls during the same period. Two real-time fluorescence quantitative PCR (RT-qPCR) techniques, namely stem-loop method and poly (A) tailed method, were used to detect the levels of 3′t-2′Ome-miR-21-5p and 3′t-2′-Ome miR-125a-5p in serum EV of NSCLC patients and controls.Analyze the correlation between the levels of two types of 3′t-2 ′Ome miRNAs and the differences in clinical stage, pathological classification, and other tumor indicators in patients. Receiver operating characteristic (receiveroperating curve, ROC) curves were used to analyze the efficacy of 3′t-2 ′Ome miR-21-5p and 3′t-2′ Ome miR-125a-5p in serum EV, as well as their combination, in diagnosing NSCLC.Result:Compared with the control group, the levels of 3′t-2′Ome-miR-21-5p in serum EV of NSCLC patients increased [(0.30±0.05) vs (0.35±0.09), t=3.32, P=0.001], while the levels of 3′t-2′Ome-miR-125a-5p decreased [(0.33±0.06 vs 0.25±0.06, t=7.45, P<0.001]. The differences were statistically significant. The expression levels of 2′Ome-miR-21-5p in EV were also significantly elevated in the NSCLC patients at 0-Ⅱ stage, adenocarcinoma patients, and squamous cell carcinoma patients, respectively. Notably, the levels of EV 3′t-2′Ome-miR-21-5p was also statistically significant between the adenocarcinoma patients and squamous cell carcinoma patients [(0.34±0.85) vs (0.40±0.12), P<0.05]. ROC curve analysis showed that the levels of 3′t-2 ′Ome miR-21-5p and 3′t-2′ Ome miR-125a-5p in serum EV, as well as their combined AUC for discriminating NSCLC patients, were 0.647(95% CI 0.550-0.743), 0.825(95% CI 0.756-0.894) and 0.860(95% CI 0.797-0.923), respectively. The sensitivity was 92.3%, 80.0%, 89.2%, and the specificity was 46.4%, 73.9%, and 78.3%, respectively. Conclusion:There are changes in the levels of 2 ′Ome miR-21-5p and 2′ Ome miR-125a-5p in the serum EV of NSCLC patients, and the combined detaction has the potential as an auxiliary screening molecular marker of NSCLC patients.

Background Type 2 diabetes mellitus is one of the most prevalent diseases,which imposes a heavy burden on patients' families and the society.Sleep disorders are recognized as risk factors for the development of diabetes,which may affect the onset and development of diabetes through neuro-endocrino-metabolic pathways,so identifying the factors responsible for the sleep quality of diabetic patients is of great importance in improving their sleep quality.Objective To investigate the relationship among fear of disease progression,executive function and sleep quality in patients with type 2 diabetes mellitus,so as to provide references for improvement of sleep quality in patients with type 2 diabetes mellitus.Methods A sample of 197 patients with type 2 diabetes mellitus who were admitted to the Endocrinology Department of the Second Affiliated Hospital of the Air Force Military Medical University from January to May 2023 and met the criteria defined in the Guideline for the Prevention and Treatment of Type 2 Diabetes Mellitus in China(2020 edition)were consecutively selected.All subjects were assessed using Fear of Progression Questionnaire-Short Form(FoP-Q-SF),Behavior Rating Inventory of Executive Function-Adult version(BRIEF-A)and Pittsburgh Sleep Quality Index(PSQI).Then the Process macro for SPSS(Model 4)and Bootstrap technique were applied to examine the mediating effect of executive function on the relationship between fear of disease progression and sleep quality in patients with type 2 diabetes mellitus.Results ①75 patients(38.07%)with type 2 diabetes mellitus were found to have sleep problems.②PSQI score in patients with type 2 diabetes mellitus was positively correlated with FoP-Q-SF score and BRIEF-A score(r=0.159,0.287,P<0.01).③Executive function mediated the relationship between fear of disease progression and sleep quality,the indirect value was 0.076(95%CI:0.022～0.146),accounting for 39.58%of the total effect.Conclusion Sleep disorders are common in patients with type 2 diabetes mellitus,and executive function may play a medicating role in the relationship between fear of disease progression and sleep quality.

Objective:To investigate the clinicopathological and molecular genetic characteristics of Crohn′s disease (CD).Methods:A retrospective analysis was conducted on 52 CD patients who underwent surgical resection at the First Affiliated Hospital of Nanjing Medical University between January 2014 and June 2023. Clinical presentations and histopathological features were assessed. Whole-genome sequencing was performed on 17 of the samples, followed by sequencing and pathway enrichment analyses. Immunohistochemistry was used to assess the expression of frequently mutated genes.Results:Among the 52 patients, 34 were males and 18 were females, male-to-female ratio was 1.9∶1.0, with a median age of 45 years at surgery and 35 years at diagnosis. According to the Montreal classification, A3 (51.9%,27/52), B2 (61.5%, 32/52), and L3 (50.0%,26/52) subtypes were the most predominant. Abdominal pain and diarrhea were the common symptoms. Histopathological features seen in all 52 patients included transmural inflammation, disruption of cryptal architecture, lymphoplasmacytic infiltration, varying degrees of submucosal fibrosis and thickening, increased enteric nerve fibers and neuronal proliferation. Mucosal defects, fissure ulcers, abscesses, pseudopolyps, and adenomatous proliferation were also observed in 51 (98.1%), 38 (73.1%), 28 (53.8%), 45 (86.5%), and 28 (53.8%) cases, respectively. Thirty-one (59.6%) cases had non-caseating granulomas, and 3 (5.8%) cases had intestinal mucosal glandular epithelial dysplasia. Molecular analysis showed that 12/17 CD patients exhibited mutations in at least one mucin family gene (MUC2, MUC3A, MUC4, MUC6, MUC12, MUC17), and MUC4 was the most frequently mutated in 7/17 of cases. Immunohistochemical stains showed reduced MUC4 expression in epithelial cells, with increased MUC4 expression in the epithelial surface, particularly around areas of inflammatory cell aggregation; and minimal expression in the lower half of the epithelium.Conclusions:CD exhibits diverse clinical and pathological features, necessitating a comprehensive multidimensional analysis for diagnosis. Mutations and expression alterations in mucin family genes, particularly MUC4, may play crucial roles in the pathogenesis of CD.

Objective To examine the expression pattern of miR-135a-5p and miR-135b-5p in seminal plasma as well as the clinical value of their variation in the male patients with infertility.Methods The clinical samples of 5 kinds,including serum,morning urine,pleural effusion,cerebrospinal fluid and seminal plasma,were collected from 30 patients respectively in the Department of Clini-cal Laboratory,Jiangsu Province Hospital of Chinese Medicine from January 2019 to December 2021.In addition,the frozen seminal plasma samples from the patients with non-obstructive azoospermia,asthenozoospermia of 54 cases for each illness statuses,and 54 healthy controls with fertile ability from 2010 to 2019 in Jiangsu Province Hospital of Chinese Medicine and Jinling Hospital were also selected.Fluorescent quantitative RT-qPCR analyses were applied to measure the expression levels of miR-135a-5p and miR-135b-5p in seminal plasma.Receiver operating characteristic curve(ROC)was used to construct the area under the curve(AUCROC)to evalu-ate the ability for discriminating infertility males from healthy controls.Correlation analyses were performed to explore the correlations between the levels of miR-135a-5p and miR-135b-5p in seminal plasma and other semen parameters.Results The concentrations of miR-135a-5p and miR-135b-5p in the samples from five different kinds of body fluid showed significantly difference(H=64.88,P＜0.01 and H=64.7,P＜0.01,respectively).The highest levels were found in serum and seminal plasma,and the lowest levels were in u-rine.The concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were also showed marked difference among the azoosper-mia group,asthenozoospermia group and healthy fertile control group(H=32.14,P＜0.01 and H=21.37,P＜0.01,respectively).Compared to the fertile control group,the levels of miR-135a-5p and miR-135b-5p in seminal plasma were significantly downregulated in azoospermia patients(U=687,P＜0.01 and U=843,P＜0.01,respectively).The contents of the two miRNAs showed no statistical-ly difference between asthenozoospermia patients and healthy fertile controls,but showed markedly difference between the two patients groups(U=635,P＜0.01 and U=779,P＜0.01,respectively).Receiver operating characteristic curve(ROC)analyses results re-vealed that levels of miR-135a-5p and miR-135b-5p in seminal plasma could successfully discriminate the azoospermia patients from fertile controls and asthenozoospermia patients with the AUCROC of 0.764(95％CI:0.675 to 0.854),0.711(95％CI:0.612 to 0.810)and 0.782(95％CI:0.695 to 0.869),and 0.733(95％CI:0.637 to 0.829),respectively.Correlation analyses demonstrated that the concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were significantly associated with sperm concentration,total sperm motility,and percentages of progressive sperm and non-progressive sperm(all with the P-value＜0.01).Conclusion miR-135a-5p and miR-135b-5p are the highly expressed miRNA in seminal plasma,and significantly decreased in azoospermia patients.Both of them may play pivot roles in male infertility.

Objective:To observe the effect of supplementing vacuum sealing drainage with hyperbaric oxygen in the short term treatment of diabetic foot ulcers.Methods:A total of 156 persons diagnosed with diabetic foot ulcers were randomly divided into a control group and a treatment group, each of 78. Both groups received life guidance and active treatment to lower blood sugar and lipids, as well as anti-infection treatment guided by bacterial cultures. Both groups′ wounds were debrided. The wound was then covered with foam, sealed, and negative pressure of -75 to -100mmHg was applied during 1 week of drainage. Two courses of this treatment were applied. In addition, the treatment group received hyperbaric oxygen daily during the two weeks. The exposure pressure was incrased to 0.25MPa over 15min with 100% oxygen. That was inhaled in two 30min sessions with a 10min interval. The pressure then decompressed at a constant rate for 25 minutes. Wound healing, hemorheology, wound granulation tissue staining and any changes in TGF-β1 were observed before as well as after 7 and 14 days of the treatment.Results:The average wound size and symptom score of both groups had improved significantly after the treatment, with the largest effect in the treatment group during the first week. Both groups′ hemorheology had improved significantly after one week, but the treatment group′s improvement was greater. After 2 weeks, however, there was no significant difference in the average hemorheologic indicators for either group compared with before the treatment. Hematoxylin-eosin staining of the wound tissues showed that there were many inflamed cells before the treatment, with relatively little fresh granulation tissue or new blood vessels. After one week of treatment much new granulation tissue was observed under the microscope in both groups, with no significant difference between them. One week later, there was still much granulation tissue in the control group, but slightly less in the treatment group. The ave-rage post-treatment TGF-β1 protein levels in the wound tissues of both groups were significantly higher than before the treatment, but after two weeks the average TGF-β1 protein level had decreased significantly in the treatment group compared with the control group.Conclusions:One week of hyperbaric oxygen treatment can effectively improve the hemorheology of persons with diabetic foot ulcers, promote the proliferation of granulation tissue and fibroblasts, and increase the level of TGF-β1 protein in the wound tissues. However, the effects of hyperbaric oxygen treatment weaken gradually with time.

Objective:To explore the application effect of segmented teaching combined with Kirkpatrick model evaluation in the teaching of specialized nurses of peripherally implanted central venous catheter (PICC).Methods:A total of 60 trainees in the PICC specialized nurse training class in PICC Clinic of Cancer Hospital Affiliated to Nantong University from June 2020 to September 2020 were selected in the study, and they were randomly divided into a control group and a study group, with 30 nurses in each group. The control group conducted traditional training for PICC specialized nurses, and the research group used segmented teaching. After training, the Kirkpatrick model was used to evaluate the response level, learning level, behavior level and result level of the training effect of the two groups of PICC specialized nurses. SPSS 22.0 was used for t test and chi-square test. Results:Through the segmented teaching training, the evaluation results of the Kirkpatrick model showed that the effect of the PICC specialized nurse training was significantly better in the study group than that in the control group in terms of response level, learning level, behavior level and result level. The study group's satisfaction with the training program and teaching content scored higher than that of the control group, with statistically significance ( P<0.001). The research group nurses' mastery of theoretical knowledge and practical skills and humanistic care scores were higher than those of the control group, with significant differences ( P<0.001). After the training, the self-behavior assessment scores of the two groups increased, and the improvement of the research group was more obvious. When the nurses of the research group returned to the hospital clinical work after training, their professional identity, PICC teaching and guidance ability, treatment, nursing service quality, the recognition of patient and family members and the effectiveness of the department were improved to varying degrees, the corresponding measures could be carried out in the hospital, and the development of PICC technology was promoted. Conclusion:The combination of segmented teaching and Kirkpatrick model evaluation can effectively meet the training needs of PICC specialized nurses and make a full range of effect evaluations to improve the quality of PICC specialized nursing services.

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL). Methods: A total of 12 specimens were collected, which were surgically resected and verified as MEITL by postoperative pathology, immumohistochemical staining and gene rearrangement at the First Affiliated Hospital of Nanjing Medical University from 2012 to 2018, and all of these had complete clinical and pathological data. The MEITL cases were reviewed to compare the clinicopathological characteristics, including morphologic and immunophenotypic features and followed up by telephone and clinic visit. Results: All the cases were diagnosed with MEITL. There were 8 males and 4 females. Male to female ratio was 2∶1, at a median age of 54 years. The sites of involvement included jejunum (4 cases), ileum (5 cases), duodenum (1 case), ileocecal junction (1 case) and rectum (1 case). The neoplastic cells were monotonous of small to intermediate cells in size with round to slightly irregular nuclei in 11 cases. The immunophenotyping showed that CD3 (12/12), CD8 (11/12), CD43 (11/12), CD56 (11/12), TIA-1 (12/12) were positive; CD5 (12/12), Gran B (9/12), and perforin (7/12) were negative. Two cases aberrantly expressed the B-cell marker CD20. A high proliferation index was demonstrated by Ki-67 immunostaining. In situ hybridization for EBER was all negative(12/12). The whole exome sequencing(WES) mutational landscape of MEITL was remarkably homogeneous, showing significantly enriched clusters among histone modifier genes, JAK-STAT and MAPK-signal pathways. Histonelysine N-methytransferase SETD2 gene was mutated in 2/4 tumors. All the patients analyzed harbored at least one mutation in the JAK-STAT signal pathway, including STAT5B (2/4), JAK3 (3/4) and STAT5A (2/4). Furthermore, frequent alterations (TP53) were observed in the MAPK pathway in 3/4 of MEITL cases. The CNV analysis derived from WES data identified multiple regions of frequent gains and losses. In particular, gains in 1q, 7q and 9q, and recurrent losses involving 7p and 8p were observed. Conclusions MEITL is a rare and aggressive type of extranodal T-cell lymphoma. The differential diagnosis of MEITL includes EATL, extranodal NT/T-cell lymphoma and other types of PTCL. Diagnosis should be correlated to clinical symptoms while the final diagnosis is mainly based on the pathological features, immunophenotypes and genetic testing.

Objective: To investigate the levels of serum piR-015151 in patients with brucellosis and evaluate its clinical value in the auxiliary diagnosis of brucellosis. Methods: Serum samples and clinical data from 73 brucellosis patients diagnosed in the Affiliated Hospital of Inner Mongolia Medical University or Zhungeer Qi Center for Disease Control and Prevention and 65 age- and gender-matched healthy controls in the Eastern Theater General Hospital of PLA during 2016 and 2017 were collected. Serum piR-015151 levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The diagnostic value of serum piR-01515 in brucellosis was evaluated by the receiver operating characteristic (ROC) curve. Results: Serum piR-015151 levels (median \[P25, P75\]) in brucellosis patients (0.216 \[0.069, 0.426\]) were significantly higher than that in healthy controls (0.036 \[0.021, 0.070\], U=834.5,P＜0.01). The area under the ROC curve (AUCROC), sensitivity and specificity of serum piR-015151 in the diagnosis of brucellosis were 0.824 (95% CI: 0.751-0.897), 75.3% and 76.9%, respectively. Conclusion Serum piR-015151 levels in brucellosis patients increase significantly, which may be a potential marker in the auxiliary diagnosis of brucellosis.

Objective: To investigate the levels of urinary exosomal miR-194-5p in children with idiopathic nephrotic syndrome (INS) and its clinical value as a non-invasive auxiliary diagnostic marker. Methods: Urine samples were collected from 101 INS children and 98 sex- and age-matched healthy children, and the levels of urinary exosomal miR-194-5p were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The clinical value of urinary exosomal miR-194-5p in the diagnosis of children′s INS was evaluated by the ROC curve and correlation analysis. Results: The levels of urinary exosomal miR-194-5p in INS children (2.420 [0.650,9.515] fmol/L) were significantly higher than that in healthy controls (0.360 [0.220,0.653] fmol/L, U=1 552, P＜0.01). Compared with the INS children before treatment, the levels of urinary exosomal miR-194-5p in clinical remission period decreased significantly (0.320 [0.145,0.523] fmol/L vs 0.975 [0.375,4.358] fmol/L, W=708, P＜0.01). Moreover, the levels of miR-194-5p in INS children with heavy urine protein (8.430 [7.225,13.070] fmol/L) were significantly higher than that with light urine protein (2.130 [1.180,3.090] fmol/L, U=0, P＜0.01). The area under the ROC curve (AUC ROC ) of miR-194-5p was 0.843 (95%CI: 0.789-0.897) for the diagnosis of INS children. Spearman correlation analysis showed that the levels of urinary exosomal miR-194-5p in INS children were negatively correlated with serum albumin levels (r=-0.300), and were positively correlated with serum total cholesterol levels (r=0.278) and 24-hour urine protein content (r=0.296, all P＜0.01). Conclusion The levels of urinary exosomal miR-194-5p in INS children increase obviously, and are closely associated with 24-hour urine protein, serum albumin and total cholesterol levels, indicating that urinary exosomal miR-194-5p may be serve as a new non-invasive fluid biopsy molecular marker for the auxiliary diagnosis of INS in children.