Aim To investigate the effects of Mdivi-1 on A1 astrocyte activation and its associated signaling molecules.Methods CTX-TNA2 astrocytes were di-vided into the control,ACM,and low-,medium-,and high-dose Mdivi-1 groups based on concentration screening via CCK-8 assay.ACM,a DMEM high-glu-cose medium containing preset concentrations of IL-1α,TNF-α,and C1q,was used to induce A1 activa-tion.The ACM group was stimulated with ACM for 24 hours.Mdivi-1 groups were pretreated with correspond-ing concentrations of Mdivi-1 for 2 hours,followed by ACM stimulation for 24 hours.Real-time quantitative PCR and Western blot were employed to assess mRNA levels and protein expression of IL-1β,C3,and iNOS in all groups.Immunofluorescence and Western blot were used to detect the expression of signaling molecules NLRP3,caspase-1,and ASC.DHE labeling was used to assess and flow cytometry was used to examine reac-tive oxygen species(ROS)levels.Results The CCK-8 assay identified 5,10,and 25 μmol·L-1as ap-propriate concentrations for Mdivi-1 intervention in CTX-TNA2 cells.Real-time quantitative PCR and Western blot results indicated that,compared to the control group,IL-1 β,C3,and iNOS mRNA levels and protein expression were significantly elevated in the ACM group(P＜0.05).In contrast,these levels were significantly reduced in the 10 and 25 μmnol·L-1 Mdi-vi-1 groups compared to the ACM group(P＜0.05).Immunofluorescence and Western blot results confirmed that ACM stimulation significantly activated the NLRP3 inflammasome in astrocytes,while Mdivi-1 intervention effectively reversed the ACM-induced upregulation of NLRP3,caspase-1,and ASC.DHE staining results demonstrated that 5,10,and 25 μmol·L-1Mdivi-1 in-terventions partially reversed the ACM-induced in-crease in ROS levels in a dose-dependent manner.Conclusion Mdivi-1 effectively inhibits A1 astrocyte activation,potentially through modulation of ROS and the NLRP3 inflammasomes.

Aim To investigate the effects of Mdivi-1 on A1 astrocyte activation and its associated signaling molecules.Methods CTX-TNA2 astrocytes were di-vided into the control,ACM,and low-,medium-,and high-dose Mdivi-1 groups based on concentration screening via CCK-8 assay.ACM,a DMEM high-glu-cose medium containing preset concentrations of IL-1α,TNF-α,and C1q,was used to induce A1 activa-tion.The ACM group was stimulated with ACM for 24 hours.Mdivi-1 groups were pretreated with correspond-ing concentrations of Mdivi-1 for 2 hours,followed by ACM stimulation for 24 hours.Real-time quantitative PCR and Western blot were employed to assess mRNA levels and protein expression of IL-1β,C3,and iNOS in all groups.Immunofluorescence and Western blot were used to detect the expression of signaling molecules NLRP3,caspase-1,and ASC.DHE labeling was used to assess and flow cytometry was used to examine reac-tive oxygen species(ROS)levels.Results The CCK-8 assay identified 5,10,and 25 μmol·L-1as ap-propriate concentrations for Mdivi-1 intervention in CTX-TNA2 cells.Real-time quantitative PCR and Western blot results indicated that,compared to the control group,IL-1 β,C3,and iNOS mRNA levels and protein expression were significantly elevated in the ACM group(P＜0.05).In contrast,these levels were significantly reduced in the 10 and 25 μmnol·L-1 Mdi-vi-1 groups compared to the ACM group(P＜0.05).Immunofluorescence and Western blot results confirmed that ACM stimulation significantly activated the NLRP3 inflammasome in astrocytes,while Mdivi-1 intervention effectively reversed the ACM-induced upregulation of NLRP3,caspase-1,and ASC.DHE staining results demonstrated that 5,10,and 25 μmol·L-1Mdivi-1 in-terventions partially reversed the ACM-induced in-crease in ROS levels in a dose-dependent manner.Conclusion Mdivi-1 effectively inhibits A1 astrocyte activation,potentially through modulation of ROS and the NLRP3 inflammasomes.

Objective To explore the clinicopathological characteristics,prognosis and influencing factors of different pathological subtypes of gastric signet ring cell carcinoma(GSRC).Methods A retrospective analysis was performed on the clinical data of 232 patients with GSRC collected from January 2016 to December 2018 in Lanzhou University Second Hospital.According to the WHO classification criteria for GSRC,the patients were divided into pure gastric signet-ring cell carcinoma(pGSRC,n=36)and mixed gastric signet-ring cell carcinoma(mGSRC,n=196).The follow-up as of September 30,2022,the survival analysis was done using Kaplan-Meier method,the univariate and multivariate Cox regression were performed to analyze the risk factors affecting the prognosis of GSRC patients.Results The median survival time of pGSRC and mGSRC patients was 41.0(6.0-70.0)months and 24.0(2.0-74.0)months,respectively.Kaplan-Meier analysis showed that combination with diabetes,anemia,tumor diameter,nerve invasion,lymphovascular invasion,T stage,N stage,GSRC pathological subtype,CA125 and tumor diameter could affect the overall survival(OS)of patients with GSRC after radical gastrectomy(P＜0.05),but Her-2,whether adjuvant chemotherapy or not and others elements had no significant effect on OS of GSRC patients after radical gastrectomy(P＞0.05).Univariate Cox regression analysis showed that the combination with diabetes(P=0.031),anemia(P=0.028),tumor diameter＞5 cm(P=0.009),nerve invasion(P=0.002),lymphovascular invasion(P=0.002),mGSRC pathological type(P=0.039),T2-T4 stage(P=0.001),N1-N4 stage(P=0.004),pTNM stage Ⅲ(P=0.044),the number of lymph node metastasis＞30(P=0.044)and CA125 positive(P=0.009)were related to the prognosis of patients with GSRC after radical gastrectomy.Multivariate Cox regression analysis showed that mGSRC pathological type(P=0.035),T2-T4 stage(P=0.003),CA125 positive(P=0.010)were independent risk factors for poor prognosis of patients with GSRC after radical gastrectomy.Conclusion Compared with pGSRC,patients with mGSRC at diagnosis have higher pTNM stages,more aggressive,and shorter median survival time.mGSRC pathological type,T2-T4 stage,and CA125 positive were all independent factors affecting the prognosis of patients with GSRC.

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.
Male ; Mice ; Animals ; Keratin-18 ; Actins ; Desmin ; Liver ; Hepatocytes ; Hepatic Stellate Cells

Transparency Ecosystem for Research and Journals in Medicine (TERM) working group summarized the essential recommendations that should be considered to review and publish a high-quality guideline. These recommendations from editors and reviewers included 10 components of essential requirements: systematic review of existing relevant guidelines, guideline registration, guideline protocol, stakeholders, conflicts of interest, clinical questions, systematic reviews, recommendation consensus, guideline reporting and external review. TERM working group abbreviates them as PAGE (essential requirements for Publishing clinical prActice GuidelinEs), and recommends guideline authors, editors, and peer reviewers to use them for high-quality guidelines.
Humans ; Practice Guidelines as Topic

ObjectiveTo explore the mechanism of Liu Junzitang in preventing and treating muscle atrophy in mice with lung cancer cachexia based on the signal transducer and activator of transcription 3（STAT3）/ubiquitin proteasome pathway in vivo. MethodForty C57BL/6 mice aged six weeks were randomly divided into a blank group， a model group， a Liu Junzitang group， an inhibitor group （stattic group），and a Liu Junzitang + inhibitor group （combination group）， with eight mice in each group. The cachectic muscle atrophy model was induced by subcutaneous inoculation of Lewis lung cancer cell line under the right anterior armpit in mice except those in the blank group. On the 8^th day after subcutaneous inoculation， the mice in the corresponding groups received Liu Junzitang （9.56 g·kg^-1·d^-1） by gavage and intraperitoneal injection of stattic ［25 mg·kg^-1·（2 d）^-1］. After three weeks of drug intervention， the body weight and gastrocnemius muscle weight were recorded. Hematoxylin-eosin （HE） staining was used to observe the pathological changes and cross-sectional area of gastrocnemius muscle fibers in mice. Western blot was used to detect the expression of phosphorylated-STAT3 （p-STAT3）， STAT3， muscle atrophy F-box （MAFbx）， and muscle RING finger protein 1 （MuRF1） in the gastrocnemius muscle. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of STAT3， MAFbx， and MuRF1 in the gastrocnemius muscle. ResultCompared with the blank group， the model group showed lightened body and the gastrocnemius muscle， reduced cross-sectional area of gastrocnemius muscle fibers， and increased protein expression of p-STAT3， STAT3， MAFbx， and MuRF1 and mRNA expression of STAT3， MuRF1， and MAFbx in the gastrocnemius muscle （P<0.05）. Compared with the model group， the Liu Junzitang group showed increased body weight， gastrocnemius muscle weight， and cross-sectional area of gastrocnemius muscle fibers （P<0.05）， and decreased protein expression of p-STAT3， STAT3， MuRF1， MAFbx， and mRNA expression of STAT3 and MAFbx in gastrocnemius muscle （P<0.05）. Compared with the model group， the inhibitor group showed increased body weight and cross-sectional area of gastrocnemius muscle fibers （P<0.05）， and reduced protein expression of p-STAT3， STAT3， MuRF1， MAFbx， and mRNA expression of STAT3 and MAFbx in gastrocnemius muscle （P<0.05）. Compared with the model group， the combination group showed increased body weight and gastrocnemius muscle weight （P<0.05），and decreased protein expression of p-STAT3， STAT3， MuRF1， and mRNA expression of STAT3 and MAFbx in the gastrocnemius muscle （P<0.05）. Compared with the Liu Junzitang group， the stattic group and the combination group showed reduced expression of p-STAT3 protein in the gastrocnemius muscle （P<0.05）. ConclusionLiu Junzitang can prevent and treat muscle atrophy in mice with lung cancer cachexia， and its mechanism may be associated with the protein and mRNA expression related to the STAT3-mediated ubiquitin proteasome pathway.

Asians ; China ; Humans ; Hypertension/drug therapy* ; Practice Guidelines as Topic ; Writing

Objective: To investigate whether the contralateral normal external auditory canal (EAC) skin graft can maintain the ear canal health after EAC reconstruction in unilateral congenital aural atresia (CAA) cases. Methods: A Zelen design randomized controlled study was used to collect unilateral CAA patients for EAC reconstruction prospectively (clinical trial registration number: ChiCTR2000032103). The patients were randomly divided into the control group and the trial group. The trial group used the contralateral normal EAC skin graft group (transplant part of the contralateral normal EAC skin to repair the atresia side for unilateral CAA patients), the control group all used scalp blade thick skin. We observed the EAC health and hearing results of the two groups after EAC reconstruction. Results: A total of 13 cases were enrolled from July 2020 to August 2021. There were eight patients in the trial group, including six males and two females, with an average age of 22.3 years (14-36 years). There were two patients with CAA on the left and six patients on the right. The average follow-up time was 8.8 months (4-14 months). There were five patients in the control group, all cases were male with an average age of 16.2 years (12-20 years). There were four patients with CAA on the left and one patient on the right. The average follow-up time was 7.0 months (2-14 months). In the trial group, eight cases of reconstructed EAC epithelium were healthy, one patient had cicatricial stenosis of EAC opening and lateralization of the tympanic membrane. The other patient had cicatricial stenosis of reconstructed EAC, this case also had scar hyperplasia of the contralateral EAC opening but recovered after soft packing and triamcinolone acetonide injection treatment. The healthy side EAC of the rest trial group had no scarring stenosis or local bone hyperplasia during long-term follow-up. In the control group, one patient was lost to follow-up and the other four patients had dry ears of reconstructed EAC, but easily to form crusts and needed to be cleaned repeatedly, one patient had lateralization of the tympanic membrane, the EAC epithelium was not healthy for long-term follow-up. The incidence of complications related to EAC reconstruction was lower than previous studies (χ²=5.55, P=0.018), and the average postoperative Air-Bone Gap increased (18.8±10.0)dB. Conclusion: By optimizing the EAC reconstruction technology, the health of the reconstructed EAC is improved compared with the previous study. After active intervention and treatment, there should be no scarring stenosis or local bone hyperplasia on the contralateral side EAC.
Adolescent ; Adult ; Ear/surgery* ; Ear Canal/surgery* ; Female ; Humans ; Male ; Retrospective Studies ; Skin Transplantation ; Tympanoplasty ; Young Adult

OBJECTIVES: At present, there are many studies on Crohn's disease of terminal ileum and colon, but few studies on Crohn's disease of small intestine alone. This study aims to analyze the clinical features and therapeutic effect of small bowel in adult patients with Crohn's disease so as to strengthen the diagnosis and treatment for this disease. METHODS: From July 1, 2015 to October 31, 2018, patients with small bowel Crohn's disease at Department of Gastroenterology, Third Xiangya Hospital of Central South University, were enrolled. At the same time, patients' demographics and clinical data were collected. RESULTS: A total of 44 patients were small bowel Crohn's disease. Among them, 40 patients were male. The age at diagnosis was (35.8±10.3) years old and disease duration was (35.2±59.5) months. The subtypes included 29(65.9%) of ileum, 7(15.9%) of jejunum, 8(18.2%) of ileum with jejunum. There were 27(61.4%) of stricture behavior, 4(9.1%) of penetrating behavior, and 13(29.5%) of non-stricture and non-penetrating behavior. Endoscopic visible stricture (29/85, 34.1%) was common, followed by longitudinal ulcers (27/85, 31.8%). Non-caseate granulomatous were found in 2 cases (4.5%). The score of Crohn's disease activity index was correlated to hemoglobin, hematocrit, and erythrocyte sedimentation rate (all CONCLUSIONS Patients with ileum account for a large proportion in patients with small bowel Crohn's disease. Stricture is more common in small bowel Crohn's disease. Stricture and longitudinal ulcer are more common under enteroscopy. Crohn's disease activity index is correlated to hemoglobin, red blood cell specific volume, and erythrocyte sedimentation rate. The most common complication is intestinal obstruction. Mesalazine is less effective on small bowel Crohn's disease.
Adult ; Crohn Disease/drug therapy* ; Endoscopy, Gastrointestinal ; Humans ; Ileum ; Intestinal Obstruction/etiology* ; Intestine, Small ; Male ; Middle Aged

Objective:The incidence rate of cancer cachexia is high in late stage of cancer， which is characterized by skeletal muscle atrophy and rapid reduction of adipose tissue and systemic inflammation. Cachexia is highly related to a variety of tumors， and causes a large proportion of cancer deaths. Cancer cachexia can lead to serious complications in patients with cancer， then， the quality of life of patients decreases， the psychological state becomes negative， and the state of illness is further worsened. At present， there is no effective intervention means to completely reverse cachexia. The combined use of multiple targets and effective components of traditional Chinese medicine（TCM）， as well as the treatment based on syndrome differentiation and the theory of TCM play an important role in the prevention and treatment of cancer cachexia. Therefore， exploring the pathogenesis of cancer cachexia and prevention and treatment with TCM is helpful for basic study and clinical application. Method:In this paper， cancer cachexia and TCM in China national knowledge infrastructure （CNKI） and Public Medline （PubMed） databases were retrieved， and 98 Chinese and English literatures were included through summarization to elaborate the pathogenesis of cancer cachexia and the prevention and treatment of TCM. Result:Emphasis was given to the important role of inflammatory reaction， skeletal muscle atrophy， energy metabolism abnormality and multiple signal joint regulation in occurrence of cancer cachexia， and the unique advantages and significant role of TCM in treatment of cancer cachexia under different treatment principles. Conclusion:Inflammatory reaction， skeletal muscle atrophy， abnormal energy metabolism make the pathogenesis of cancer cachexia complex and diverse. TCM prescriptions， Chinese herbal medicine and their effective ingredients have the natural advantages of targeting multiple pathways， controlling multiple signal pathways and inhibiting various inflammatory factors in the prevention and treatment of cancer cachexia， and are safe and effective in improving diet， prolonging the survival period of patients and keeping weight.