OBJECTIVES: To analyze the differentially expressed exosomal miRNAs in patients with chronic heart failure (CHF) complicated by hyperuricemia (HUA) and explore their potential as novel diagnostic molecular markers and their target genes. METHODS: This study was conducted among 30 CHF patients with HUA (observation group) and 30 healthy volunteers (control group) enrolled between September, 2020 and September, 2023. Peripheral blood samples were collected from 6 CHF patients with HUA for analyzing exosomal miRNAs by high-throughput sequencing, and the results were validated in the remaining 24 patients using qRT-PCR. GO and KEGG enrichment analyses were performed to predict the the target genes of the identified differential miRNAs. We also validated the differentially expressed miRNAs by animal experiment. RESULTS: A total of 42 differentially expressed exosomal miRNAs were detected in observation group by high-throughput sequencing; among them, miR-27a-5p was significantly upregulated (P=0.000179), and miR-139-3p was significantly downregulated (P=0.000058). In the 24 patients with both CHF and PUA, qRT-PCR validated significant upregulation of miR-27a-5p (P=0.004) and downregulation of miR-139-3p (P=0.005) in serum exosomes. When combined, miR-27a-5p and miR-139-3p had a maximum area under the curve (AUC) of 0.899 (95% CI: 0812-0.987) for predicting CHF complicated by HUA. GO and KEGG enrichment analyses suggested that the differential expressions of miR-27a-5p and miR-139-3p was associated with the activation of the AMPK-mTOR signaling pathway to activate the autophagic response. We obtained the same conclusion from animal experiment. CONCLUSIONS Upregulated exosomal miR-27a-5p combined with downregulated exosomal miR-139-3p expression can serve as a novel molecular marker for diagnosis of CHF complicated by HUA, and their differential expression may promote autophagy in cardiomyocytes by activating the AMPK-mTOR signaling pathway.
Humans ; Hyperuricemia/diagnosis* ; Heart Failure/genetics* ; MicroRNAs/metabolism* ; Exosomes/metabolism* ; Chronic Disease ; Male ; Female ; Middle Aged ; Animals

OBJECTIVES: To explore the bioactive components in Jiawei Xiaoyao Pills (JWXYP) and their mechanisms for alleviating depression-like behaviors. METHODS: The active compounds, key targets, and pathways of JWXYP were identified using TCMSP and TCMIP databases. Thirty-six SD rats were randomized equally into 6 groups including a control group and 5 chronic unpredictable mild stress (CUMS)-induced depression groups. After modeling, the 5 model groups were treated with daily gavage of normal saline, 1.8 mg/kg fluoxetine hydrochloride (positive control drug), or JWXYP at 1.44, 2.88, and 4.32 g/kg. The depression-like behaviors of the rats were evaluated using behavioral tests, and pathological changes in the liver and hippocampus were examined with HE staining. The biochemical indicators in the serum and brain tissues were detected using ELISA. Serum metabolomics analysis was performed to identify the differential metabolites using OPLS-DA, and gut microbiota changes were analyzed using 16S rDNA sequencing. RESULTS: Network pharmacology revealed that menthone and paeonol in JWXYP were capable of penetrating the blood-brain barrier to regulate inflammatory pathways and protect the nervous system. In the rat models subjected to CUMS, treatment with JWXYP significantly improved body weight loss, sucrose preference and open field activities, reduced liver inflammation, alleviated structural changes in the hippocampal neurons, decreased serum levels of TNF‑α, IL-1β, IL-6 and LBP, and increased 5-HT and VIP concentrations in the serum and brain tissue, and these effects were the most pronounced in the high-dose group. Metabolomics analysis showed changes in such metabolites as indole-3-acetamide and acetyl-L-carnitine in JWXYP-treated rats, involving the pathways for bile acid biosynthesis and amino acid metabolism. 16S rDNA analysis demonstrated increased gut microbiota diversity and increased abundance of Lactobacillus species in JWXYP-treated rats. CONCLUSIONS JWXYP alleviates depression-like symptoms in rats by regulating the neurotransmitters, inhibiting inflammation and oxidation, and modulating gut microbiota.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Gastrointestinal Microbiome/drug effects* ; Rats, Sprague-Dawley ; Depression/drug therapy* ; Neurotransmitter Agents/metabolism* ; Rats ; Inflammation ; Male ; Hippocampus ; Behavior, Animal/drug effects*

Objective To explore the effect of circular RNA(circRNA)transcription factor 25(TCF25)-targeting microRNA-128b(miR-128b)on the proliferation and apoptosis of MCF-7 human breast cancer cells.Methods MCF-7 cells were cultured until the loga-rithmic growth stage.Cells were randomly divided into the blank(untransfected),si-NC(transfected si-NC),si-circRNA TCF25(transfected si-circRNA TCF25),pcDNA-circRNA TCF25(transfected pcDNA-circRNA TCF25),miR-NC(transfected miR-NC),miR-128b mimic(transfected miR-128b mimic),miR-128b inhibitor(transfected miR-128b inhibitor),and pcDNA-circRNA TCF25+miR-128b mimic(transfected with pcDNA-circRNA TCF25 and miR-128b mimic)groups.Each group included six multiple pores.Forty-eight hours after transfection,the expression of circRNA TCF25 and miR-128b in each group was determined using real-time reverse transcription-quanti-tative polymerase chain reaction(RT-qPCR).An RNase R enzyme digestion assay was used to identify circular RNA.Subcellular locali-zation of circRNA TCF25 was determined through cytoplasmic-nuclear separation.Cell proliferative activity was measured using the 2,5-diphenyl-2H-tetrazolium bromide(MTT)assay.Apoptosis was detected using flow cytometry.RT-qPCR was performed to determine the mRNA expression levels of phosphatase and tensin homolog(PTEN),proliferating nuclear antigen 67(Ki-67),andcaspase-3.Western blotting was performed to measure PTEN,Ki-67,caspase-3,and cleaved caspase-3 protein expression.The dual-luciferase reporter(DLR)assay was performed to analyze the relationship between circRNA TCF25 and miR-128b.Results Compared to the control group,the relative expression of circRNA TCF25 did not exhibit significant changes after RNase R enzyme treatment(P＞0.05),whereas that of linear TCF25 decreased after RNase R enzyme treatment(P＜0.05).The relative expression of circRNA TCF25 in the cytoplasm was higher than that in the nucleus(P＜0.05).Compared with the blank and si-NC groups,the cell proliferation activity of the si-circRNA TCF25 group decreased;the apoptosis rate increased;Ki-67mRNA and protein expression decreased;and PTEN,caspase-3,and cleaved caspase-3mRNA and protein expression increased.In addition,cell proliferation activity increased and apoptosis rate decreased in the pcDNA-circRNA TCF25 group.Ki-67 mRNA and protein expression were increased,and PTEN,caspase-3,and cleaved caspase-3 mRNA and protein expression decreased,with statistical significance(all P＜0.05).Compared with the blank and miR-NC groups,the cell proliferation activity of the miR-128b mimic group decreased;the apoptosis rate increased;Ki-67mRNA and protein expression decreased;and PTEN,caspase-3,and cleaved caspase-3 mRNA and protein expression were increased,whereas the cell proliferation activity increased and apoptosis rate decreased in the miR-128b inhibitor group.Ki-67mRNA and protein expression were increased,and PTEN,caspase-3,and cleavedcaspase-3mRNA and protein expression decreased,with statistical significance(all P＜0.05).Compared with the pcDNA circRNA TCF25 group,the cell proliferation activity of the pcDNA circRNA TCF25+miR-128b mimic group decreased;the apoptosis rate increased;Ki-67mRNA and protein expression decreased;and PTEN,caspase-3,and cleavedcaspase-3mRNA and protein expression increased,with statistical significance(all P＜0.05).The DLR assay results confirmed that circRNA TCF25 targets miR-128b.Conclusion CircRNAs may play a key role in promoting the proliferation of MCF-7 human breast cancer cells and inhibiting their apoptosis by targeting miR-128b expression;promoting Ki-67 expression;and inhibiting PTEN,caspase-3,and cleaved caspase-3 expression.

ObjectiveTo explore the meridian tropism of components in Bupleuri Radix （Chaihu， CH） based on the model of nonalcoholic steatohepatitis （NASH） and clarify the substance basis of the meridian tropism of CH in Xiaoyaosan （XYS） by means of principal component analysis. MethodEighty SPF male C57BL/6 mice were randomly assigned into 8 groups， with 10 mice in each group. Except that the blank group was fed with the methionine choline-sufficient （MCS） diet， the other mice were fed with methionine choline-deficient （MCD） diet for 4 weeks to establish the nonalcoholic steatohepatitis （NASH） model. After the established model was confirmed by hematoxylin-eosin （HE） staining， the mice were administrated with corresponding drugs by gavage once a day for 4 weeks. Specifically， the 8 groups were XYS group （2.874 g·kg^-1）， XYS-CH group （2.445 g·kg^-1）， XYS-CH+volatile oils （Vol， 0.163 mg·kg^-1） group， XYS-CH+polysaccharides （Pol， 24.067 mg·kg^-1） group， XYS-CH+flavones （Fla， 2.241 mg·kg^-1） group， and XYS-CH+saponins （Sap， 2.746 mg·kg^-1） group. The model group and the blank group were administrated with the same volume of normal saline. After the last administration， the mice were sacrificed for the collection of blood and liver tissue. The pathological changes of liver were observed by HE staining and oil red O staining. Enzyme linked immunosorbent assay （ELISA） kits were used to determine the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein （HDL）， and low-density lipoprotein （LDL） in serum as well as malondialdehyde （MDA）， superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） in liver. SPSS Statistics 23 was used for principal component analysis and comprehensive evaluation to determine the substance basis of the meridian tropism of CH in NASH mice. ResultCompared with the blank control group， the modeling led to hepatocyte swelling， increased fat vacuoles， and appearance of inflammatory cells. Further， the modeling elevated the levels of ALT， AST， TG， TC， and LDL and lowered the HDL level in serum， and it increased the MDA level and decreased the SOD， CAT， and GSH-Px levels in liver. Compared with the model group， the administration of XYS and XYS-CH in combination with the components of CH alleviated the oxidative damage in liver （P<0.05）. The comprehensive score of the pharmacological efficacy was in a descending order as follows： XYS > XYS-CH+Sap > XYS-CH+Fla > XYS-CH+Pol > XYS-CH+Vol > XYS-CH. Among the chemical components of CH， Sap had the best effect. ConclusionSap lowers the blood lipid level， regulates the abnormal lipid metabolism， and alleviates the oxidative damage of liver， which is the substance basis for CH to exert the meridian tropism in liver.

Objective:To compare and analyze the differences in the setup accuracy of different immobilization method in breast cancer radiotherapy after breast-conserving surgery.Methods:A retrospective study was conducted on 60 patients who received radiotherapy after breast-conserving surgery from January to August, 2021. These patients were divided into two groups. One group consisted of 30 cases who were immobilized using a modified body thermoplastic membrane combined with a multifunction body board during the breast cancer radiotherapy and was called the modified body thermoplastic membrane group. The other group comprised 30 cases immobilized using a vacuum cushion during breast cancer radiotherapy and was referred to as the vacuum cushion group. The setup errors, 3D vector errors, the proportion of errors of > 5 mm, and the dosimetric differences in the planning target volume (PTV) and the clinical target volume (CTV) before and after simulated treatment bed moving (including the PTV_ V100, PTV_ V95, and CTV_ V95 before simulated treatment bed moving and the PTV_ V100 S, PTV_ V95 S, and CTV_ V95 S after simulated treatment bed moving) were compared between two groups. Moreover, for the modified body thermoplastic membrane group, the changes in the average setup errors at different radiotherapy stages were also analyzed. Results:A total of 369 cone-beam CT scans were conducted for 60 patients, including 195 CT scans for the modified body thermoplastic membrane group and 174 CT scans for the vacuum cushion group. The setup errors in the x, y, and z directions (right-left, anterior-posterior, and superior-inferior, respectively) of the modified body thermoplastic membrane group were (2.59±1.98) mm, (2.38±2.04) mm, and (1.45±1.16) mm, respectively, while those of the other group were (2.24±1.63) mm, (2.78±2.17) mm, and (2.70±1.88) mm, respectively. The 3D vector errors of both groups were (4.32±2.28) mm and (5.13±2.14) mm, respectively. Therefore, the setup error in direction z and the 3D vector error of the modified body thermoplastic membrane group were less than those of the vacuum cushion group ( t = -7.77, -3.41, P<0.05). Moreover, the proportion of setup errors of > 5 mm in the x direction of the vacuum cushion group was lower than that of the modified body thermoplastic membrane group ( χ2 = 7.13, P<0.05), while such proportion in the z direction of the modified body thermoplastic membrane group was lower than that of the vacuum cushion group ( χ2= 5.90, P<0.05). After the simulated treatment bed moving, the PTV_ V100 S of the modified body thermoplastic membrane group was better than that of the vacuum cushion group ( t = 2.47, P < 0.05). Furthermore, for the modified body thermoplastic membrane group, the setup errors in the x direction in the first week were higher than those in the 2-3 weeks and 4-5 weeks ( P<0.05). Conclusions:The modified body thermoplastic membrane combined with a multifunction body board yield better immobilization effects than a vacuum cushion. However, it produces high setup errors in the x direction in the first week of the radiotherapy, to which special attention should be paid.

Tocilizumab has been reported to attenuate the "cytokine storm" in COVID-19 patients. We attempted to verify the effectiveness and safety of tocilizumab therapy in COVID-19 and identify patients most likely to benefit from this treatment. We conducted a randomized, controlled, open-label multicenter trial among COVID-19 patients. The patients were randomly assigned in a 1:1 ratio to receive either tocilizumab in addition to standard care or standard care alone. The cure rate, changes of oxygen saturation and interference, and inflammation biomarkers were observed. Thirty-three patients were randomized to the tocilizumab group, and 32 patients to the control group. The cure rate in the tocilizumab group was higher than that in the control group, but the difference was not statistically significant (94.12% vs. 87.10%, rate difference 95% CI-7.19%-21.23%, P = 0.4133). The improvement in hypoxia for the tocilizumab group was higher from day 4 onward and statistically significant from day 12 (P = 0.0359). In moderate disease patients with bilateral pulmonary lesions, the hypoxia ameliorated earlier after tocilizumab treatment, and less patients (1/12, 8.33%) needed an increase of inhaled oxygen concentration compared with the controls (4/6, 66.67%; rate difference 95% CI-99.17% to-17.50%, P = 0.0217). No severe adverse events occurred. More mild temporary adverse events were recorded in tocilizumab recipients (20/34, 58.82%) than the controls (4/31, 12.90%). Tocilizumab can improve hypoxia without unacceptable side effect profile and significant influences on the time virus load becomes negative. For patients with bilateral pulmonary lesions and elevated IL-6 levels, tocilizumab could be recommended to improve outcome.
Antibodies, Monoclonal, Humanized ; COVID-19/drug therapy* ; Humans ; SARS-CoV-2 ; Treatment Outcome

Objective To compare the therapeutic effects between three-dimensional conformal radiotherapy (3DCRT) and intensity-modulated radiotherapy (IMRT) in patients with stage Ⅱ/Ⅲ esophageal cancer and investigate the prognostic factors.Methods Medical record of 2 132 patients with stage Ⅱ/Ⅲ esophageal cancer who underwent definitive radiotherapy with/without chemotherapy in 10 hospitals from January 2002 to December 2016 from were retrospectively analyzed.Among these patients,37.9％ of them were aged ≥ 70 years,33.9％ with neck and upper esophageal tumors and 66.1％ with middle and lower esophageal and borderline tumors.The median gross tumor volume (GTV) and lymph node gross tumor volume (GTVnd) was 41.6 cm3.Among them,32％ were stage Ⅱ] and 68％ were stage Ⅲ.A total of 723 patients received 3DCRT and 1 409 cases received IMRT.Patients received an equivalent dose in 2 Gy (EQD2) ≥ 60 Gy accounted for 86.1％,and 41.1％ of them received concurrent chemoradiotherapy.Results The median follow-up time was 60.8 months.The 1-,3-and 5-year overall survival (OS) of all patients was 73.9％,41.7％ and 32.6％,and the 1-,3-and 5-year progression-free survival (PFS) was 62.2％,37.3％ and 32％,respectively.Multivariate analysis demonstrated that age,primary tumor location,clinical stage,tumor target volume,EQD2 and concurrent chemoradiotherapy were the independent prognostic factors for OS.Age,primary tumor location,clinical stage,tumor target volume and EQD2 were the independent prognostic factors for PFS.The OS and PFS did not significantly differ among the low-risk,low-/moderate-risk,moderate-/high-risk and high-risk groups according to age≥70 years,tumor diameter＞5 cm,tumor volume ≥41.6 cm3 and stage Ⅲ (P＜0.001).After the propensity score matching (PSM) method,neither 3DCRT nor IMRT yielded significant advantages in OS or PFS (P=0.971;P=0.658).However,IMRT tended to yield survival benefits in low-risk patients (P=0.125).Conclusions Both 3DCRT and IMRT yield relatively high OS rate in patients with stage Ⅱ/Ⅲ esophageal cancer.The prognosis model established in this investigation can properly predict the survival of patients.Low-risk patients tend to obtain survival benefits from IMRT.

Objective To evaluate the clinical efficacy and toxicity of intensity-modulated radiation therapy (IMRT) following prostatectomy for elderly patients with prostate cancer.Methods Ninety-eight prostate cancer patients receiving IMRT after prostatectomy were included in this study.The median age was 68 years old.The number of patients with low-,middle-and high-risk prostate cancer was 10,21 and 67,respectively.Two patients had oligometastases (pelvic bone metastases).Sixty-four patients were treated with IMRT combined with endocrine therapy.Among them,43 cases received adjuvant volumetric modulated arc therapy (VMAT),and 55 patients received salvage IMRT.The median radiotherapy dose was 72 Gy for the tumor bed.Twenty-nine patients received radiotherapy of the pelvic node region with a median dose of 50 Gy.Results The median follow-up time was 40 months.The 5-year overall survival (OS),biochemical recurrence-free survival (BRFS) and local control (LC) were 90％,76％ and 100％,respectively.The OS (88.8％ vs.90.8％,P=0.94),BRFS (75.9％ vs.71％,P=0.79) or LC (100％ vs.100％,P=0.32) did not significantly differ between the adjuvant and salvage radiotherapy groups,respectively.The incidence of grade Ⅰ-Ⅱ late rectal toxicities was 24.1％,and no ≥ grade 3 late toxicity was observed.The incidence of grade 1-2 late bladder toxicities was 29.9％,and 3.4％ for grade 3.Conclusion IMRT following prostatectomy yields high clinical efficacy and slight late toxicities in elderly patients with prostate cancer.

Objective To evaluate the clinical efficacy and adverse events of intensity-modulated radiotherapy ( IMRT ) in the treatment of intermediate risk localized prostate cancer, and analyze the significance of prostate-specific antigen ( PSA) level changes. Methods Clinical data of 66 patients with intermediate risk localized prostate cancer admitted to our hospital between 2007 and 2018 were retrospectively analyzed. Sixty patients were treated with endocrine therapy before radiotherapy. The radiation field covered the pelvic lymph node drainage area in 6 cases. Forty-seven patients received image-guided radiotherapy ( IGRT) . The median dose in the prostate and seminal vesicle was 78 Gy and 48 Gy in the pelvic lymph node drainage area. The survival rate was calculated using the Kaplan-Meier method. Results The median age was 77 years. The median follow-up time was 71. 3 months. The 5-year sample size was 47. The 3-and 5-year overall survival (OS) was 98％ and 90％.The 3-and 5-year cancer-specific survival (CSS) was 100％ and 93％.The 3-and 5-year biochemical relapse-free survival was 97％ and 86％. The mean time of PSA declining to the nadir was 5. 83 months. The median level of PSA nadir was 0. 06 ng/ml after IMRT. The incidence of grade I andⅡearly adverse events in the urinary system was 38％ and 6％. The incidence of grade I andⅡearly adverse events in the gastrointestinal system was 21％ and 3％. The incidence of grade I andⅡadvanced-stage adverse events in the urinary system was 9％ and 2％. The incidence of grade I advanced-stage adverse events in the gastrointestinal system was 5％. Conclusions IMRT yields high clinical efficacy in the treatment of intermediate risk localized prostate cancer with a low risk of adverse events in the early and advanced stage. The monitoring of PSA after IMRT contributes to the assessment of clinical prognosis.

Objective To analyze the effect of endoscopic middle turbinate angioplasty on postoperative nasal function and olfaction in patients with chronic sinusitis and nasal polyp.Methods Clinical materials of 80 patients with chronic sinusitis and nasal polyps were analyzed retrospectively,and they were divided into control group and research group,40 cases in each group.Control group was treated with endoscopic partial middle turbinate resection,while research group was treated with endoscopic middle turbinate angioplasty.Clinical efficacy,nasal function,olfactory function,SNOT-20CV score,and Lund-Kennedy scoring and complications were compared between two groups.Results Total effective rate of the research group was significantly higher than the control group (P ＜0.05).Before operation,nasal airway resistance and minimal nasal cross-section to anterior nostril distance (DCAN),nasal minimum cross-sectional area (NMCA),nasal volume (NCV),SNOT-20 CV,nasal endoscopic Lund-Kennedy scores showed no significant differences between two groups (P ＞ 0.05).After operation,nasal airway resistance and DCAN and SNOT-20 CV,nasal endoscopic Lund-Kennedy scores decreased significantly in both groups,and these indexes in research group were significantly lower than the control group (P ＜ O.05).The NMCA and NCV increased significantly in both groups,and these indexes in research group were significantly higher than control group (P ＜ 0.05).There was no significant difference in incidence rate of postoperative complications between the two groups (P ＞ 0.05).Conclusion Endoscopic middle turbinate angioplasty is effective in treating chronic sinusitis patients with nasal polyp,which can effectively relieve nasal obstruction and promote nasal function and the sense of smell.