Prognostic validity of the American joint committee on cancer eighth edition staging system for well-differentiated pancreatic neuroendocrine tumors

BackgroundThe American Joint Committee on Cancer (AJCC) made improvements for staging pancreatic neuroendocrine tumors (pNETs) in its 8th Edition; however, multicenter studies were not included.MethodsWe collected multicenter datasets (n = 1,086, between 2004 and 2018) to validate the value of AJCC 8 and other coexisting staging systems through univariate and multivariate analysis for well-differentiated (G1/G2) pNETs.ResultsCompared to other coexisting staging systems, AJCC 7 only included 12 (1.1%) patients with stage III tumors. Patients with European Neuroendocrine Tumor Society (ENETS) stage IIB disease had a higher risk of death than patients with stage IIIA (hazard ratio [HR]: 4.376 vs. 4.322). For the modified ENETS staging system, patients with stage IIB disease had a higher risk of death than patients with stage III (HR: 6.078 vs. 5.341). According to AJCC 8, the proportions of patients with stage I, II, III, and IV were 25.7%, 40.3%, 23.6%, and 10.4%, respectively. As the stage advanced, the median survival time decreased (NA, 144.7, 100.8, 72.0 months, respectively), and the risk of death increased (HR: II = 3.145, III = 5.925, and IV = 8.762).ConclusionThese findings suggest that AJCC 8 had a more reasonable proportional distribution and the risk of death was better correlated with disease stage.     

Identification of differentially expressed miRNAs and key genes involved in the progression of alcoholic fatty liver disease using rat models

BackgroundAlcoholic fatty liver disease (AFLD) is a liver disease caused by prolonged heavy drinking and has a poor prognosis in the clinic. This study aimed to explore the differential miRNAs expression profiles in the AFLD rat model.MethodsThe rat model of AFLD was established by ethanol intragastric administration and was used to explore the differential miRNAs expression profiles. We further analyzed the potential target mRNAs using the bioinformatics technique. GO and KEGG pathway enrichment analyses were carried out to better understand the biological function of differential expression genes (DEGs). We used the human Gene Expression Omnibus (GEO) dataset GSE28619 to further screen the key differentially expressed genes. The integration between the differentially expressed genes from the AFLD model and GEO was conducted and the key genes were identified.ResultsThe serum ALT, AST, TG, and TC levels in the AFLD model group were significantly higher than those in the normal control group. There are 45 miRNAs with significant changes including 26 upregulated and 19 down-regulated miRNAs. GO and KEGG enrichment showed various metabolic processes and signaling pathways were enriched in the progression of AFLD. After integrating the results of GSE28619 and DEGs, we observed that there are 12 genes with significant changes in two data sets, including PSAT1, TKFC, PTTG1, LCN2, CXCL1, NR4A1, RGS1, VCAN, FOS, CXCL10, ATF3, and CYP1A1.ConclusionAFLD showed differentially expressed miRNAs, which may be involved in the occurrence and progression of AFLD. Meanwhile, some signal metabolic pathways may be related to the pathogenesis of AFLD.     

Bone Metabolism Discrepancy in Type 2 Diabetes Mellitus Patients With and Without Non-Alcoholic Fatty Liver Disease

To explore the distribution of several bone metabolic indicators in type 2 diabetes patients (T2DM) with and without non-alcoholic fatty liver disease (NAFLD) and to preliminarily evaluate the relationship of bone metabolism with NAFLD in patients with T2DM. The hospitalized patients with T2DM were divided into the group of T2DM complicated with NAFLD and the group of T2DM alone according to the results of ultrasonic diagnosis. The general information and laboratory test data such as bone metabolism indexes of these patients were collected and the differences of the indexes between the 2 groups were compared. Furthermore, the independent influencing factors of NAFLD in patients with T2DM were analyzed. A total of 186 patients were included in the study. Compared with patients with T2DM only, patients with T2DM combined with NAFLD were characterized with younger age (p < 0.001), higher BMI (p = 0.016), ALT (p = 0.001), TG (p = 0.005), HOMA-IR (p = 0.005), and lower HDL-C (p = 0.031). Significant discrepancy of age (OR 1.052, p = 0.001), ALT (OR 0.964, p = 0.047), HOMA-IR (OR 0.801, p = 0.005), and T-PINP (OR 1.022, p = 0.008) was found using multivariate logistic regression model. Significant discrepancy of T-PINP was found in T2DM patients with and without NAFLD. Further studies are needed to explore whether T-PINP could be used as a predictor of fatty liver disease, osteoporosis, and other related complications in patients with T2DM.     

血浆血管内皮细胞生长因子与肝细胞癌临床特征的相关性

目的通过检测肝细胞癌(HCC)患者肝动脉栓塞术前血浆血管内皮细胞生长因子(VEGF),考察VEGF表达与肿瘤大小、边界、门脉癌栓、远隔转移之间的相关性。方法收集介入治疗前HCC患者46例,VEGF定量检测采用双抗体夹心酶ABC ELISA法。根据血浆VEFG表达水平将患者分为阳性组(32例)和阴性组(14例),对肿瘤大小、边界、门脉癌栓、远隔转移情况进行组间对比。CT和DSA间接门脉造影获得的图像进行对比评估。结果VEGF阳性表达率69.56%(32/46),阴性表达率30.44%(14/46)。VEGF阳性组肿瘤直径>5cm者26例(81.25%),阴性组12例(85.75%),两组间无显著性差异(P>0.05)。CT片上阳性组肿瘤边界清楚15例(46.88%),阴性组边界清楚8例(57.14%),组间无显著性差异。阳性组门脉癌栓发生率62.50%(20/32),阴性组14.29%(2/14),两组间有极显著性差异(P<0.01)。阳性组远隔转移8例;阴性组无远隔转移,经检验P<0.05。结论血浆VEGF高表达可以提示门脉癌栓或远隔转移的存在,而与肿瘤大小、边界无关。血浆VEGF可能成为观察肿瘤门脉癌栓及远隔转移的指标。     

Activation of the Notch signaling pathway disturbs the CD4+/CD8+, Th17/Treg balance in rats with experimental autoimmune uveitis

Objective and design

The present study aimed to investigate the relationship between the disturbed balance of CD4⁺/CD8⁺, Th17/Treg and the activation of the Notch signaling pathway in experimental autoimmune uveitis (EAU).

Methods

An EAU rat model was induced in Lewis rats, and pathology analysis was performed by hematoxylin and eosin (H&E) staining. CD4⁺, CD8⁺, Th17, and Treg levels in spleen, lymph nodes and eye tissues were determined by flow cytometry. Meanwhile, the expression of Notch1, DLL4, IL-10, and IL-17 was determined by quantitative polymerase chain reaction (Q-PCR) and enzyme-linked immunosorbent assay (ELISA). In addition, the inhibitory effect of N-(N-(3,5-difluorophenacetyl-l-alanyl))-S-phenylglycine t-butyl ester (DAPT) on Th17 differentiation by Notch signaling in vitro was further investigated using T lymphocytes from EAU rats on day 12 post-immunization by flow cytometry.

Results

The pathological results showed that inflammatory cell infiltration occurred in ocular tissues in EAU rats. The CD4⁺/CD8⁺ and Th17/Treg ratios in EAU rats were apparently higher than those in normal control individuals. Q-PCR and ELISA analyses indicated the expression of Notch1, DLL4, IL-10, and IL-17 in EAU rats gradually increased on day 6 after immunization, peaked on day 12, and then gradually decreased. The dynamic trends in Notch1 and DLL4 expression in EAU rats were identical to those of CD4⁺/CD8⁺ and Th17/Treg levels. DAPT can significantly inhibit the activation of Notch signaling, decrease Th17 cell differentiation, and attenuate the level of the Th17 cell lineage, contributing to the balance of the Th17/Treg ratio.

Conclusion

The activation of the Notch signaling pathway can regulate Th17 and Treg cell differentiation, disrupt the CD4⁺/CD8⁺ and Th17/Treg balance, and aggravate the severity of EAU; inactivation of the Notch signaling pathway contributes to the CD4⁺/CD8⁺ and Th17/Treg balance in EAU rats. Our findings highlighted that the dynamic change in the CD4⁺/CD8⁺ and Th17/Treg ratio was consistent with the expression trend of Notch signaling in EAU rats, suggesting that Notch signaling may be a potentially important therapeutic target in clinical practice.

     

Study of electroacupuncture with different frequencies for the treatment of headache and anxiety-depression symptoms in patients with migraine: A randomized clinical trial: 不同频率电针改善偏头痛患者头痛及焦虑抑郁症状的差异性研究-随机临床试验

ObjectiveTo observe the effects of electroacupuncture (EA) with varied frequencies on headache and anxiety-depression symptoms in patients with migraine and to screen optimal frequency of EA for patients with such conditions.DesignSingle-center, randomized, controlled clinical trial was designed, and the outcome assessors and statisticians were blinded. The patients with migraine were randomized into 2 Hz EA group, 100 Hz EA group and sham-stimulation group. In each group, the changes in migraine attacks, days with headache, the scores of visual analogy scale (VAS), self-rating anxiety scale (SAS), self-rating depression scale (SDS) and migraine-specific quality-of-life questionnaire (MSQ), as well as the dosage of analgesics were observed at the baseline, during treatment, in 1, 2 and 3 months of follow-up separately.SettingPatients were enrolled in the Third Affiliated Hospital of Zhejiang Chinese Medical University, between 1^st August 2018 and 31^st July 2021.ParticipantsSixty-five migraine patients with or without aura.InterventionsIn the EA groups, the acupoints were bilateral Fengchi (GB20), Gongxue (Extra), Sizhukong (TE23), Taiyang (EX-HN5), Shuaigu (GB8), Waiguan (TE5) and Yanglingquan (GB34). Electric stimulation was exerted at GB20 and Gongxue (Extra), with 2 Hz and 100 Hz separately. In the sham-stimulation group, the shallow acupuncture was operated at the sites 1 cm lateral to GB20 and Gongxue (Extra), and on the radial side of TE5 and GB34. The output wires were cut off after attached to the acupoints. The patients in each group received the treatment 3 times weekly, once every two days, for consecutive 4 weeks. The complete intervention was composed of 12 treatments.Main outcome measuresChanges in numbers of migraine attacks at treatment phase (week 1 to week 4) from the baseline(week -4 to week 0) in patients of each groupResults(1) Changes in migraine attacks and days with headache: In the 2 Hz EA and 100 Hz EA groups, the changes for migraine attacks and days of headache were higher significantly when compared with that in the sham-stimulation group at the same time stage (P < 0.05). There was no statistical difference between two EA groups. (2) Changes of VAS score: In the 2 Hz EA and 100 Hz EA groups, the changes of VAS score were significantly higher when compared with that in the sham-stimulation group at the same time stage (P < 0.05). There was no statistical difference between two EA groups. (3) Assessment of anxiety and depression: The differences in the changes of SAS and SDS scores had no statistical significance in patients of each group at each assessment stage (P > 0.05). (4) Assessment on the quality of life: Compared with the sham-stimulation group at the same time stage, the improvement in MSQ score was increased significantly during treatment in patients of the 2 Hz EA and 100 Hz groups (P < 0.05). There was no statistical difference between two EA groups. (5) Assessment on safety and compliance: The patients of each group had sound compliance. There was no adverse events during trial, suggesting promising safety of treatment.ConclusionEA may effectively reduce the migraine attacks, and the days and intensity of headache, presenting promising safety. However, there was no significant improvement on anxiety-depression symptoms, and no significant difference between high and low frequencies of EA treatment in relieving headache and anxiety/depression symptoms in the patients with migraine.Trial registrationChiCTR1800017259     

Clinical Characteristics and Controllable Risk Factors of Osteoporosis in Elderly Men with Diabetes Mellitus

ObjectiveTo analyze the clinical characteristics and controllable risk factors of osteoporosis in elderly men with type‐2 diabetes mellitus (T2DM).MethodsA total of 250 elderly OP patients with T2DM were included in the present study. Patients with one or more common chronic diseases (including hypertension, coronary heart disease, heart failure, chronic bronchitis, chronic nephrosis, and cirrhosis), and a course of more than 3 years were defined as complicated with chronic diseases. Blood glucose, cholesterol, triglyceride, low‐density lipoprotein, high‐density lipoprotein, calcium, phosphorus, glycosylated hemoglobin, urea nitrogen, creatinine, fasting insulin, liver function, and 25‐hydroxy vitamin D3 levels were measured. Bone mineral density was also measured.ResultsA total of 16 patients (6.4%) had severe osteoporosis. Furthermore, 66 patients (26.4%) had blood glucose control that reached the standard, while 176 patients (70.4%) used more than two anti‐diabetic drugs. The serum testosterone level was lower than the median in 87 patients (34.8%) and in 56 smokers (22.4%). Furthermore, 138 patients (55.2%) were overweight and obese, six patients (2.4%) were underweight, 197 patients (78.8%) had chronic diseases, 88 patients (35.2%) were sticking to exercise, and 117 patients (46.8%) had less exercise. In addition, 92 patients (36.8%) were treated with osteotrophy‐protective agents, and 24 patients (9.6%) received anti‐osteoporosis therapy. Smoking, poor glycemic control, low testosterone levels, less exercise, and complications with chronic diseases were the most relevant controllable risk factors.ConclusionFor elderly male osteoporosis patients with type‐2 diabetes, smoking cessation, blood sugar control up to the standard, regular exercise, active prevention and treatment of complications, and appropriate testosterone supplementation are necessary for preventing and curing osteoporosis.     

miR-146a-5p regulates autophagy and NLRP3 inflammasome activation in epithelial barrier damage in the in vitro cell model of ulcerative colitis through the RNF8/Notch1/mTORC1 pathway

Ulcerative colitis (UC) is a chronic inflammatory disease affecting the colon that can be influenced by microRNAs (miRNAs). This study aims to investigate the impact of miR-146a-5p on lipopolysaccharide (LPS)-induced Caco-2/HT-29 cell autophagy and NLRP3 inflammasome activation and the underlying mechanism, with the aim of identifying potential therapeutic targets. We used LPS to establish Caco-2/HT-29 cell models and measured cell viability by CCK-8. The levels of miR-146a-5p, RNF8, markers of NLRP3 inflammasome activation and autophagy, proteins involved in the Notch1/mTORC1 pathway, and inflammatory factors were assessed by RT-qPCR, Western blot, and ELISA. Intestinal epithelial barrier function was evaluated by measuring transepithelial electrical resistance. Autophagic flux was measured using tandem fluorescent-labeled LC3. miR-146a-5p was highly-expressed in LPS-induced Caco-2/HT-29 cells, and autophagy flux was blocked at the autolysosomal stage after LPS induction. Inhibition of miR-146a-5p suppressed NLRP3 inflammasome activation, reduced intestinal epithelial barrier damage, and facilitated autophagy inhibition in LPS-induced Caco-2/HT-29 cells. The autophagy inhibitor NH4Cl partially nullified the inhibitory effects of miR-146a-5p inhibition on NLRP3 inflammation activation. miR-146a-5p targeted RNF8, and silencing RNF8 partly abrogated the action of miR-146a-5p inhibition on promoting autophagy and inhibiting NLRP3 inflammasome activation. miR-146a-5p inhibition suppressed the Notch1/mTORC1 pathway activation by upregulating RNF8. Inhibition of the Notch1/mTORC1 pathway partially nullified the function of silencing RNF8 on inhibiting autophagy and bolstering NLRP3 inflammasome activation. In conclusion, miR-146a-5p inhibition may be a potential therapeutic approach for UC, as it facilitates autophagy of LPS-stimulated Caco-2/HT-29 cells, inhibits NLRP3 inflammasome activation, and reduces intestinal epithelial barrier damage by upregulating RNF8 and suppressing the Notch1/mTORC1 pathway.