AIM:This study examined the inhibitory effect of peiminine on the human colonic adenocarcino-ma cell line SW480 and explored the underlying mechanisms.METHODS:SW480 and human normal colonic epithelial CCD-841CoN cells were treated with different concentrations of peiminine and subjected to the CCK-8 assay to select the optimal treatment time and concentration of the compound.SW480 cell migration and invasion were evaluated by the wound-healing and Transwell assays.Cell cycle progression was analyzed by flow cytometry.The expression levels of cell cycle-related proteins were examined by Western blot.SW480 xenograft tumor model was established in nude mice to ex-amine the effect of peiminine on tumor growth and the expression of cell cycle-related proteins in vivo.RESULTS:Peimi-nine(110 mg/L)significantly inhibited the proliferation of SW480 cells compared with the control group(P<0.01),caused cell cycle arrest at G1 phase,and significantly downregulated the expression of cyclin dependent kinase 2(CDK2),CDK4,CDK6,cyclin D1,p-Rb/Rb,E2F1,E2F3,and E2F4(P<0.05).Peiminine inhibited SW480 xenograft tumor growth,prolonged the survival of model mice,and affected the expression of CDK2,CDK4,CDK6,and cyclin D1 in tu-mor tissues.CONCLUSION:Peiminine promotes G1 phase arrest by down-regulating the expression of CDK2,CDK4,CDK6,and cyclin D1,thereby inhibiting the proliferation of SW480 cells.

Objective:To explore the diagnostic value of magnetic resonance(MR)high-resolution imaging of vascular wall on vertebrobasilar artery tortuosity prolongation complicated by multiple cerebral infarction in posterior circulation.Methods:Regression analysis was used in this research.A total of 138 patients with vertebrobasilar artery tortuosity prolongation who admitted to Hengshui City People′s Hospital from January 2020 to January 2023 were selected.Among of them,54 patients who complicated with multiple cerebral infarction in posterior circulation were divided into the occurrence group,and 84 patients without multiple cerebral infarction in posterior circulation were divided into the non-occurrence group.All patients underwent high-resolution MR imaging of vascular wall,and the basilar artery(BA),hemodynamic indicators such as systolic peak(Vs),average peak(Vd),diastolic peak flow velocity(Vm),pulsatile index(PI),blood flow resistance index(RI),as well as intracranial responsibility plaque characteristics and vertebral basilar artery grading between two groups were compared.Results:The Vs,Vd,Vm,PI and RI indicators of patients in the non-occurrence group were significantly higher than those in the occurrence group,and the differences were statistically significant(t=11.742,4.323,7.367,4.824,2.351,P<0.05),respectively.The degree(60.31±4.24)%of intracranial plaque stenosis in the non-occurrence group was significantly lower than(73.67±5.96)%in the occurrence group,and the difference was statistically significant(t=15.376,P<0.05).There were no statistically significant differences in plaque area,plaque load,arterial remodeling index and eccentricity index between the two groups(P>0.05).The height level of vertebral basilar artery bifurcation and the degree of vertebral basilar artery deviation in the non-occurrence group were significantly lower than those in the occurrence group,with statistically significant differences(x2=3.907,6.419,P<0.05).Conclusion:MR high-resolution imaging of vascular wall can clearly display the blood flow velocity and intracranial plaque situation in patients with vertebrobasilar artery tortuosity prolongation,and conduct the grade for the vertebrobasilar artery,which has significant application value in predicting the occurrence of vertebrobasilar artery tortuosity prolongation complicated by multiple cerebral infarction in posterior circulation.

Nucleic acid-based molecular diagnostic methods are considered the gold standard for detecting infectious pathogens.However,when applied to portable or on-site rapid diagnostics,they still face various limitations and challenges,such as poor specificity,cumbersome operation,and portability difficulties.The CRISPR(Clustered regularly interspaced short palindromic repeats)/CRISPR-associated protein(Cas)-fluorescence detection method holds the potential to significantly enhance the specificity and signal-to-noise ratio of nucleic acid detection.In this study,we developed a portable grayscale reader detection system based on loop-mediated isothermal amplification(LAMP)-CRISPR/Cas.On one hand,in the presence of CRISPR RNA(crRNA),the CRISPR/Cas12a system was employed to achieve precise fluorescent detection of self-designed LAMP amplification reactions for influenza A and influenza B viruses.This further validated the high selectivity and versatility of the CRISPR/Cas system.On the other hand,the accompanying independently developed portable grayscale reader allowed for low-cost collection of fluorescence signals and high-reliability visual interpretation.At the end of the detection process,it directly provided positive or negative results.Practical sample analyses using this detection system have verified its reliability and utility,demonstrating that this system can achieve highly sensitive and highly specific portable analysis of influenza viruses.

【Objective】 To compare the degree, efficacy and safety of testicular catch-up growth after inguinal and subinguinal surgery for adolescent varicocele. 【Methods】 Clinical data of 121 adolescent varicocele patients with secondary testicular atrophy treated during Sep.2014 and Jun.2021 at Huili People’s Hospital were retrospectively analyzed, including 40 cases treated with the inguinal incision approach, and 81 with the subinguinal approach. The characteristics of intraoperative, efficacy, surgical complications and prognosis were compared between the two groups. 【Results】 The inguinal group had significantly shorter microscope usage time (P=0.006), fewer number of preserved internal spermatic arteries (P<0.001), and significantly larger diameter of preserved internal spermatic arteries than the subinguinal group (P=0.005). The subinguinal group required ligation of more internal spermatic veins, and there were significantly more internal spermatic veins with a diameter of 4 mm or more (P<0.001). No recurrence occurred, and the incidences of scrotal edema and hydrocele were very low (P>0.05). Two years after surgery, the bilateral testicular volume of both groups was significantly larger than that before surgery (P<0.05), and there was no significant difference in the testicles (P>0.05). 【Conclusion】 In the treatment of adolescent varicocele patients with secondary testicular atrophy, the subinguinal approach significantly reduces the difficulty of microsurgical varicocele ligation, and has good efficacy and safety, which is worthy of clinical promotion and application.

Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.

Objective:To investigate the current status of endoscopic treatment for gastroesophageal varices in portal hypertension in China, and to provide supporting data and reference for the development of endoscopic treatment.Methods:In this study, initiated by the Liver Health Consortium in China (CHESS), a questionnaire was designed and distributed online to investigate the basic condition of endoscopic treatment for gastroesophageal varices in portal hypertension in 2022 in China. Questions included annual number and indication of endoscopic procedures, adherence to guideline for preventing esophagogastric variceal bleeding (EGVB), management and timing of emergent EGVB, management of gastric and isolated varices, and improvement of endoscopic treatment. Proportions of hospitals concerning therapeutic choices to all participant hospitals were calculated. Guideline adherence between secondary and tertiary hospitals were compared by using Chi-square test.Results:A total of 836 hospitals from 31 provinces (anotomous regions and municipalities) participated in the survey. According to the survey, the control of acute EGVB (49.3%, 412/836) and the prevention of recurrent bleeding (38.3%, 320/836) were major indications of endoscopic treatment. For primary [non-selective β-blocker (NSBB) or endoscopic therapies] and secondary prophylaxis (NSBB and endoscopic therapies) of EGVB, adherence to domestic guideline was 72.5% (606/836) and 39.2% (328/836), respectively. There were significant differences in the adherence between secondary and tertiary hospitals in primary prophylaxis of EGVB [71.0% (495/697) VS 79.9% (111/139), χ2=4.11, P=0.033] and secondary prophylaxis of EGVB [41.6% (290/697) VS 27.3% (38/139), χ2=9.31, P=0.002]. A total of 78.2% (654/836) hospitals preferred endoscopic therapies treating acute EGVB, and endoscopic therapy was more likely to be the first choice for treating acute EGVB in tertiary hospitals (82.6%, 576/697) than secondary hospitals [56.1% (78/139), χ2=46.33, P<0.001]. The optimal timing was usually within 12 hours (48.5%, 317/654) and 12-24 hours (36.9%, 241/654) after the bleeding. Regarding the management of gastroesophageal varices type 2 and isolated gastric varices type 1, most hospitals used cyanoacrylate injection in combination with sclerotherapy [48.2% (403/836) and 29.9% (250/836), respectively], but substantial proportions of hospitals preferred clip-assisted therapies [12.4% (104/836) and 26.4% (221/836), respectively]. Improving the skills of endoscopic doctors (84.2%, 704/836), and enhancing the precision of pre-procedure evaluation and quality of multidisciplinary team (78.9%, 660/836) were considered urgent needs in the development of endoscopic treatment. Conclusion:A variety of endoscopic treatments for gastroesophageal varices in portal hypertension are implemented nationwide. Participant hospitals are active to perform emergent endoscopy for acute EGVB, but are inadequate in following recommendations regarding primary and secondary prophylaxis of EGVB. Moreover, the selection of endoscopic procedures for gastric varices differs greatly among hospitals.

Objective To predict the target and molecular mechanism of Huayu Xiaopi decoction in the intervention of Precancerous lesions of gastric cancer(PLGC)based on network pharmacology and molecular docking technology,and to conduct experimental verification.Methods A total of 60 SPF SD male rats were randomly selected as blank control,and the other rats were replicated in PLGC model.After successful modeling,the rats were randomly divided into model group,folic acid group(2 mg·kg-1·d-1),Huayu Xiaopi decoction high,medium and low dose groups(24.8,12.4,6.2 g·kg-1·d-1),which were continuously administered for 90 days.The body mass and food intake of rats at 3 h were recorded,and the gastric histopathology was observed by HE staining.Network pharmacology and molecular docking techniques were used to predict the potential targets of Huayu Xiaopi decoction in PLGC intervention,and the core targets were verified by Western blot technique.Results Compared with the blank group,the body mass and 3 h food intake of rats in the model group were significantly decreased(P<0.05),the gastric mucosa of rats was significantly thinner,the glands were significantly reduced and disordered,and the intestinal metaplasia goblet cells and a large number of inflammatory cells were visible in some areas.Compared with the model group,the body mass and 3 h food intake of rats in each administration group were improved to varying degrees.Huayu Xiaopi Decoction improved significantly in medium and high doses(P<0.05),the gastric mucosa was repaired in different degrees,the glandular arrangement tended to be orderly,and the inflammatory cells in the interstitial were gradually reduced.The results of network pharmacology and molecular docking showed that TP53,JUN and MAPK3/1(ERK1/2)were the core targets of Huayu Xiaopi decoction in the intervention of PLGC.Molecular biological detection results showed that compared with blank group,the protein phosphorylation levels of TP53,c-Jun and ERK1/2 in gastric tissue of model group were significantly increased(P<0.05).Compared with model group,the protein phosphorylation levels of TP53,c-Jun and ERK1/2 in gastric tissue of rats in all administration groups were decreased to different degrees,and significantly decreased in Huayu Xiaopi decoction high-dose and medium-dose groups(P<0.05).Conclusion Huayu Xiaopi Decoction can significantly improve the survival condition of PLGC rats and promote gastric mucosal repair,the specific mechanism of which may be related to the decrease of ERK1/2,c-Jun and TP53 protein phosphorylation levels in gastric tissue of PLGC rats,and then regulate the downstream signaling molecular response.

Objective To observe the effect of matrine on intestinal mucosal injury in rats with inflammatory bowel disease(IBD)by regulating interleukin 6(IL-6)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods IBD model rats were constructed by colon perfusion of trinitrobenzene sulfonic acid(TNBS)method,and randomly divided into model group,matrine low-and high-dose groups,and matrine high-dose+ colivelin(STAT3 activator)group,with 10 rats in each group.Another 10 rats were selected for colon perfusion with equal volume of normal saline as the normal group.After treatment with matrine and colivelin,the body mass and disease activity index(DAI)of rats in each group were detected.The histopathological changes of colonic mucosa in rats were detected by hematoxylin-eosin(HE)staining.The ultrastructural changes of colonic mucosa in rats were observed by transmission electron microscopy.The levels of interleukin-6(IL-6),C-reactive protein(CRP),and superoxide dismutase(SOD),malondialdehyde(MDA)in serum and colonic mucosa of rats in each group were measured by enzyme-linked immunosorbent assay(ELISA)and colorimetry,respectively.The expression of IL-6/STAT3 pathway-related proteins in rat colonic mucosa was detected by Western Blot.Results Compared with the normal group,the colonic mucosal tissue of rats in the model group had severe pathological damage and obvious ultrastructural damage,DAI,colonic mucosal histopathological score,serum and colonic mucosal tissue CRP,IL-6,MDA levels,and IL-6 protein expression and p-STAT3/STAT3 levels in colonic mucosal tissue were significantly increased(P＜0.05),and body mass,serum and colonic mucosal tissue SOD level were significantly decreased(P＜0.05).Compared with the model group,the pathological damage and ultrastructural damage of colonic mucosa in the low-and high-dose of matrine groups were alleviated,DAI,pathological score of colonic mucosa,CRP,IL-6,MDA levels in serum and colonic mucosa,IL-6 protein expression and p-STAT3/STAT3 ratio in colonic mucosa were decreased(P＜0.05),and body mass,SOD level in serum and colonic mucosa were increased(P＜0.05),in a dose-dependent manner.Compared with the matrine high-dose group,the pathological damage and ultrastructural damage of colonic mucosa in the matrine high-dose + colivelin group were aggravated,DAI,colonic mucosa histopathological score,serum and colonic mucosa CRP,IL-6,MDA levels,colonic mucosa IL-6 protein expression and p-STAT3/STAT3 ratio were increased(P＜0.05),body mass,serum and colonic mucosa SOD level were decreased(P＜0.05).Conclusion Matrine can reduce the level of inflammation and oxidative stress by preventing the activation of IL-6/STAT3 signal pathway,thereby reducing intestinal mucosal injury in IBD rats.

OBJECTIVE: To explore the relationship between serum lipoprotein-associated phospholipase A2 (Lp-PLA2) level and the risk of acute ischemic stroke (AIS) recurrence in hypertensive patients. METHODS: This retrospective case-control study was conducted among 211 hypertensive patients with AIS treated in Foshan First People's Hospital, including 35 patients with recurrence of AIS during the 1-year follow-up as confirmed by head CT/MR. In the overall patients, 60 had grade 1 hypertension (including 5 recurrent cases), 76 had grade 2 hypertension (with 11 recurrent cases), and 75 had grade 3 hypertension (with 19 recurrent cases). Univariate analysis, multivariate logistic regression analysis, trend analysis, and smooth curve fitting analysis were performed to explore the correlation between serum Lp-PLA2 level within 24 h after admission and the risk of AIS recurrence. The predictive efficacy of serum Lp-PLA2 level for AIS recurrence in different hypertension grades was evaluated using ROC curve analysis. RESULTS: Serum Lp-PLA2 level, age, NIHSS score at admission, mRS scores at 7 days, homocysteine level and smoking status differed significantly between patients with and without AIS recurrence (P < 0.05). After adjustment for confounding factors, multivariate regression analysis showed that the highest tertile of Lp-PLA2 level was associated with a 4.13-fold increase of AIS recurrence risk compared with the lowest tertile (OR=5.13, 95% CI: 1.35-19.40), and each 1 ng/mL increase of Lp-PLA2 level was associated with a 1% increase of AIS recurrence risk (OR= 1.01, 95% CI: 1.01-1.02). Serum Lp-PLA2 level was shown to positively correlate with AIS recurrence risk, and in patients with grade 3 hypertension, its areas under the ROC curve for predicting AIS recurrence was 0.869 with a specificity of 0.893 and a sensitivity of 0.737. CONCLUSION Serum Lp-PLA2 concentration is an independent risk factor and potentially an effective predictor for AIS recurrence in patients with grade 3 hypertension.
Humans ; Infant, Newborn ; 1-Alkyl-2-acetylglycerophosphocholine Esterase ; Acute Disease ; Biomarkers ; Brain Ischemia/etiology* ; Case-Control Studies ; Cerebral Infarction ; Hypertension/complications* ; Ischemic Stroke/complications* ; Retrospective Studies ; Risk Factors ; Stroke

OBJECTIVE: Physical exercise, a common non-drug intervention, is an important strategy in cancer treatment, including hepatocellular carcinoma (HCC). However, the mechanism remains largely unknown. Due to the importance of hypoxia and cancer stemness in the development of HCC, the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness. METHODS: A physical exercise intervention of swimming (30 min/d, 5 d/week, for 4 weeks) was administered to BALB/c nude mice bearing subcutaneous human HCC tumor. The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring, hematoxylin and eosin (HE) staining, and immunohistochemistry (IHC) detection of proliferating cell nuclear antigen (PCNA) and Ki67. The expression of stemness transcription factors, including Nanog homeobox (NANOG), octamer-binding transcription factor 4 (OCT-4), v-Myc avian myelocytomatosis viral oncogene homolog (C-MYC) and hypoxia-inducible factor-1α (HIF-1α), was detected using real-time reverse transcription polymerase chain reaction. A hypoxia probe was used to explore the intratumoral hypoxia status. Western blot was used to detect the expression of HIF-1α and proteins related to protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway. The IHC analysis of platelet endothelial cell adhesion molecule-1 (CD31), and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion. SMMC-7721 cells were treated with nude mice serum. The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors. The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1α. Further, the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected. RESULTS: Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor. HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention. Swimming potently alleviated the intratumoral hypoxia, attenuated the cancer stemness, and inhibited the Akt/GSK-3β/β-catenin signaling pathway. Additionally, the desmin⁺/CD31⁺ ratio, rather than the number of CD31⁺ vessels, was significantly increased in swimming-treated mice. In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere, as well as the mRNA expression level of stemness transcription factors. Consistent with the in vivo results, HIF-1α and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group. CONCLUSION Swimming alleviated hypoxia and attenuated cancer stemness in HCC, through suppression of the Akt/GSK-3β/β-catenin signaling pathway. The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor. Please cite this article as: Xiao CL, Zhong ZP, Lü C, Guo BJ, Chen JJ, Zhao T, Yin ZF, Li B. Physical exercise suppresses hepatocellular carcinoma progression by alleviating hypoxia and attenuating cancer stemness through the Akt/GSK-3β/β-catenin pathway. J Integr Med. 2023; 21(2): 184-193.
Humans ; Animals ; Mice ; Carcinoma, Hepatocellular/drug therapy* ; Proto-Oncogene Proteins c-akt/metabolism* ; Proliferating Cell Nuclear Antigen/therapeutic use* ; Mice, Nude ; Glycogen Synthase Kinase 3 beta/genetics* ; beta Catenin/therapeutic use* ; Liver Neoplasms/drug therapy* ; Desmin/therapeutic use* ; Ki-67 Antigen ; Cell Line, Tumor ; Hypoxia ; RNA, Messenger/therapeutic use* ; Cell Proliferation