In recent years, gastrointestinal stromal tumors (GIST) have increased incidence and mortality, and most GIST is caused by the activation mutation of the c-KIT gene. Therefore, c-KIT has become a promising therapeutic target of GIST. At present, the drugs approved for the treatment of GIST including imatinib, sunitinib, regorafenib and ripretinib, are mostly prone to developing resistance and accompanied by various degrees of adverse reactions. Therefore, there is an urgent need to develop new c-KIT inhibitors to solve the problem of resistance. In this study, we investigated the anti-tumor effect of a novel c-KIT inhibitor PN17-1 on gastrointestinal stromal tumor GIST-882 cells in vitro. We found that PN17-1 significantly inhibited the proliferation, colony formation and migration ability of GIST-882 cells, and significantly downregulated the protein expression levels of p-c-KIT and its downstream signals p-AKT, p-STAT5 and p-ERK in GIST-882 cells. In addition, PN17-1 induced apoptosis in GIST-882 cells, and the apoptosis may be mainly related to the mitochondrial-dependent endogenous pathway. In conclusion, the novel c-KIT inhibitor PN17-1 is a promising anti-GIST drug, and this study provides new ideas for further development of c-KIT inhibitors in the future.

Objective To explore the protective mechanism of icariin on neuronal oxidative damage,providing a basic pharmacological basis for the treatment of cognitive impairment.Methods Glutamate was used to induce oxidative stress injury in HT22 cells.HT22 cells were divided into control group(normal cultured cells),model group(glutamate injury model)and experimental-L,-M,-H groups(5,10 and 20 μmol·L-1 icariin pretreatment for modeling,respectively).Cell proliferation was detected by cell counting kit-8(CCK-8)method;cytotoxicity was detected by lactate dehydrogenase(LDH)method;reactive oxygen species(ROS)levels were detected by flow cytometry;superoxide dismutase(SOD)levels were detected by biochemical kits;the expression levels of Kelch-like epichlorohydrin-related protein-1(Keap1),nuclear factor E2-related factor 2(Nrf2)were detected by Western blotting;the corresponding mRNA expression was detected by real-time fluorescence quantification polymerose chain reaction.Results The cell viability of control group,model group and experimental-L,-M,-H groups were(100.00±1.31)％,(66.38±2.44)％,(72.07±4.95)％,(82.41±3.57)％and(87.97±4.98)％;LDH release were(0.48±0.52)％,(18.82±2.09)％,(15.32±1.17)％,(10.37±1.39)％and(6.51±0.87)％;ROS level were(14.23±1.13)％,(41.74±1.60)％,(35.69±1.08)％,(33.28±1.69)％and(30.32±2.03)％;SOD levels were(54.84±1.17),(37.95±1.13),(48.02±1.28),(50.56±1.34)and(52.55±1.04)U·mg-1;Keap1 protein levels were 0.36±0.01,0.52±0.03,0.46±0.04,0.39±0.09 and 0.35±0.12;Nrf2 protein levels were 0.29±0.02,0.13±0.08,0.18±0.03,0.21±0.11 and 0.26±0.04;catalase(CAT)mRNA levels were 1.01±0.08,0.81±0.06,0.90±0.04,1.05±0.15 and 1.33±0.26;SOD mRNA levels were 1.09±0.12,0.83±0.03,0.86±0.08,0.94±0.08 and 1.09±0.16.Among the above indicators,the differences between the model group and the control group were statistically significant(all P＜0.01);the differences between the experimental-M,-H groups and the model group were statistically significant(P＜0.01,P＜0.05).Conclusion Icariin may activate the Keap1/Nrf2/antioxidant response element(ARE)signaling pathway,regulate the expression of related proteins,and reduce the level of ROS to effectively alleviate oxidative stress injury in neuronal cells.

AIM:To investigate the relationship among lens parameters and their correlation with ocular anatomic characteristics in myopia patients implanted with posterior-chamber phakic implantable collamer lens(Phakic-ICL).METHODS:Retrospective study. A total of 46 myopia patients(46 eyes)who underwent Phakic-ICL implantation were collected in the Wuxi Huaxia Eye Hospital from June 2022 to June 2023. Preoperative evaluation of ocular anatomical characteristics included corneal central thickness(CCT), anterior chamber depth(ACD), axial length(AL), white-to-white(WTW), horizontal sulcus to sulcus(STSH), horizontal angle to angle(ATAH), and vertical sulcus to sulcus(STSV), vertical angle to angle(ATAV). Furthermore, lens parameters included horizontal crystalline lens rise(CLRH), vertical CLR(CLRV)and vertical lens thickness(LTV). The difference, consistency and correlation of the above parameters were analyzed.RESULTS:Except for differences between WTW and STSV, STSH and ATAV, which were not statistically significant(all P>0.05), the other horizontal and vertical ocular anatomical characteristics were statistically significant(P<0.05). CLRH and CLRV had statistically significant difference(P<0.01), while LTH and LTV were not statistically significant difference(P>0.05). Bland-Altman results revealed that the anatomical characteristics in the horizontal or vertical diameters showed poor consistency. The consistency between CLRH and CLRV was poor. There was consistency between LTH and LTV, with the 95% limits of agreement(LoA)between the differences ranging from -0.21 to 0.28 mm, and the proportion of out-of-line points off the 95% LoA was 4.35%. Pearson correlation analysis revealed that there were correlations between the anatomical characteristics of the horizontal and vertical diameters(P<0.01). Meanwhile, there was no correlation between the anteroposterior diameters(P>0.05). There were correlations between the lens parameters(P<0.05), excepted for the CLRH, LTH and LTV, which had no correlation. AL correlated with the anatomical characteristics of the horizontal and vertical diameters(P<0.05), but it had no correlation with lens parameters(P>0.05). Multiple linear regression analysis revealed that LT=0.419+0.017×age-0.548×ACD+0.371×ATAH+0.884×CLRV, CLRH=-0.443+0.809×CLRV, CLRV=-0.092-0.200×ATAH+0.560×CLRH(corrected R2=0.458, 0.482, 0.589, respectively).CONCLUSION:Horizontal and vertical diameters were not interchangeable. CLRH and CLRV were not interchangeable, while LTH and LTV were interchangeable. Partial lens parameters, WTW, STS, and ATA were correlated with ACD. Finally, age, ACD, ATAH, and CLRV influenced LT.

Objective:To investigate the prognosis and influencing factors of endoscopic surgery for early glottic carcinoma.Methods:In this retrospective study, we applied the Cox proportional hazards regression model and the random survival forest model to analyze the clinical characteristics of 385 patients [362 males, 23 females, age ranging from 33 to 91 years (62.0±9.6)] who visited the Sixth Medical Center of the General Hospital of the People's Liberation Army from January 2009, to December 2022 and diagnosed with early glottic carcinoma, encompassing variables such as age, gender, T stage, surgical approach, pathological typing, etc. The primary evaluation indicators were overall survival(OS) and disease-free survival rates (DFS). The follow-up duration ranged from 30 to 5,557 days (with a median follow-up time of 1,596 days).Results:After a three-year follow-up, the OS rate for the 385 patients was 95.83%, while the DSF rate was 82.98%. The Cox proportional hazards regression analysis revealed age ( HR=2.35, 95% CI: 1.75 to 3.15, P<0.001) and T staging ( HR=1.59, 95% CI: 1.13 to 2.23, P=0.019) as predominant factors affecting the OS and DFS. The random survival forest model identified poor tumor differentiation, and high expression of P53 and Ki-67 as predictors of inferior prognosis. Conclusion:Endoscopic surgery for early glottic carcinoma yields favorable short-term OS and reduces short-term recurrence rates, with T-stage emerging as a pivotal factor influencing recurrence. Tumors with poor differentiation and elevated expression of P53 may be indicative of an increased risk of recurrence.

The analysis of nano-micro nuclear particles has attracted significant attention due to the crucial role of their elemental and isotopic characteristics in tracing the origins of particulate matter and assessing its potential risks to human health and the environment.However,challenges persist in obtaining accurate and consistent element profiles and ratios for small-sized nanoparticles due to their low level and the transient nature.In this study,a high-speed digital oscilloscope was integrated with multiple collector inductively coupled plasma mass spectrometry(MC-ICP-MS)to develop a high time-resolution"Event-triggered signal capture"(ETSC)system for single particle analysis.This innovative approach enabled the analysis of element/isotope within rare earth nanoparticles at ag-fg level.The ETSC accurately recorded the complete profile of single particle,event captured by the electron multiplier with nanosecond time resolution,allowing for high-sensitivity element analysis and high-precision isotope analysis of single particles.The results demonstrated that the ETSC system could achieve quantitative analysis of ag levels of ytterbium(Yb)in 50-nm rare earth-doped particles,with a detection limit as low as 38 ag for Yb.Moreover,the isotopic precision of single particle analysis for 173/171Yb could reach 0.047(standard deviation),and the standard error for isotopic analysis of multiple particles could achieve a level of 2‰-3‰(permil)for 173/171Yb.Finally,the capability of ETSC system to analyze environmental samples was demonstrated through the analysis of doped ytterbium oxide nanoparticles.All these findings demonstrated that the ETSC provided a unique method for elemental and isotopic analysis of single nuclear particles.

Vascular wall-resident stem cells (VW-SCs) play a critical role in maintaining normal vascular function and regulating vascular repair. Understanding the basic functional characteristics of the VW-SCs will facilitate the study of their regulation and potential therapeutic applications. The aim of this study was to establish a stable method for the isolation, culture, and validation of the CD34⁺ VW-SCs from mice, and to provide abundant and reliable cell sources for further study of the mechanisms involved in proliferation, migration and differentiation of the VW-SCs under various physiological and pathological conditions. The vascular wall cells of mouse aortic adventitia and mesenteric artery were obtained by the method of tissue block attachment and purified by magnetic microbead sorting and flow cytometry to obtain the CD34⁺ VW-SCs. Cell immunofluorescence staining was performed to detect the stem cell markers (CD34, Flk-1, c-kit, Sca-1), smooth muscle markers (SM22, SM MHC), endothelial marker (CD31), and intranuclear division proliferation-related protein (Ki-67). To verify the multipotency of the isolated CD34⁺ VW-SCs, endothelial differentiation medium EBM-2 and fibroblast differentiation medium FM-2 were used. After culture for 7 days and 3 days respectively, endothelial cell markers and fibroblast markers of the differentiated cells were evaluated by immunofluorescence staining and q-PCR. Furthermore, the intracellular Ca²⁺ release and extracellular Ca²⁺ entry signaling were evaluated by TILLvisION system in Fura-2/AM loaded cells. The results showed that: (1) High purity (more than 90%) CD34⁺ VW-SCs from aortic adventitia and mesenteric artery of mice were harvested by means of tissue block attachment method and magnetic microbead sorting; (2) CD34⁺ VW-SCs were able to differentiate into endothelial cells and fibroblasts in vitro; (3) Caffeine and ATP significantly activated intracellular Ca²⁺ release from endoplasmic reticulum of CD34⁺ VW-SCs. Store-operated Ca²⁺ entry (SOCE) was activated by using thapsigargin (TG) applied in Ca²⁺-free/Ca²⁺ reintroduction protocol. This study successfully established a stable and efficient method for isolation, culture and validation of the CD34⁺ VW-SCs from mice, which provides an ideal VW-SCs sources for the further study of cardiovascular diseases.
Mice ; Animals ; Endothelial Cells ; Cell Differentiation/physiology* ; Stem Cells ; Adventitia ; Fibroblasts ; Cells, Cultured ; Antigens, CD34/metabolism*

This study aimed to explore the intervention effect of Chuanxiong-Chishao herb pair(CX-CS) on a myocardial infarction-atherosclerosis(MI-AS) mouse model and investigate its effect on the expression profile of circular RNAs(circRNAs)/long non-coding RNAs(lncRNAs) in ischemic myocardium and aorta. Sixty male ApoE~(-/-) mice were randomly assigned to a model group, high-, medium-, and low-dose CX-CS groups(7.8, 3.9, and 1.95 g·kg~(-1)), and a positive drug group(metoprolol 26 mg·kg~(-1) and simvastatin 5.2 mg·kg~(-1)), with 12 mice in each group. Male C57BL/6J mice were assigned to the sham group. The mice in the model group and the groups with drug intervention were fed on a high-fat diet for 10 weeks, followed by anterior descending coronary artery ligation. After that, the mice were fed on a high-fat diet for another two weeks to induce the MI-AS model. The mice in the sham group received normal feed, followed by sham surgery without coronary artery ligation. Mice in the groups with drug intervention received CX-CS or positive drug by gavage for four weeks from the 9th week of high-fat feeding, and those in the model group and the sham group received an equal volume of normal saline. Whole transcriptome sequencing was performed on the heart and aorta tissues of the medium-dose CX-CS group, the model group, and the sham group after administration. The results showed that the medium-and high-dose CX-CS groups showed improved cardiac function and reduced myocardial fibrosis area, and the medium-dose CX-CS group showed significantly reduced plaque area. CX-CS treatment could reverse the expression of circRNA＿07227 and circRNA＿11464 in the aorta of AS model and circRNA expression(such as circRNA＿11505) in the heart of the MI model. Differentially expressed circRNAs between the CX-CS-treated mice and the model mice were mainly enriched in lipid synthesis, lipid metabolism, lipid transport, inflammation, and angiogenesis in the aorta, and in angiogenesis, blood pressure regulation, and other processes in the heart. CX-CS treatment could reverse the expression of lncRNAs such as ENSMUST00000162209 in the aorta of the AS model and TCONS＿00002123 in the heart of the MI model. Differentially expressed lncRNAs between the CX-CS-treated mice and model mice were mainly enriched in lipid metabolism, angiogenesis, autophagy, apoptosis, and iron death in the aorta, and in angiogenesis, autophagy, and iron death in the heart. In summary, CX-CS can regulate the expression of a variety of circRNAs and lncRNAs, and its intervention mechanism in coronary heart disease may be related to the regulation of angiogenesis and inflammation in ischemic myocardium, as well as lipid metabolism, lipid transport, inflammation, angiogenesis in AS aorta.
Animals ; Male ; Mice ; Atherosclerosis/genetics* ; Lipids ; Mice, Inbred C57BL ; Myocardial Infarction/genetics* ; RNA, Circular/genetics* ; RNA, Long Noncoding/genetics*

OBJECTIVE: To observe the effects of electro-scalp acupuncture （ESA） on the expression of microglial markers CD206 and CD32, as well as interleukin (IL)-6, IL-1β, and IL-10 in the ischemic cortex of rats with ischemic stroke, and to explore the mechanisms of ESA on alleviating inflammatory damage of ischemic stroke. METHODS: Sixty 7-week-old male SD rats were randomly selected, with 15 rats assigned to a sham surgery group. The remaining rats were treated with suture method to establish rat model of middle cerebral artery occlusion (MCAO). The rats with successful model were randomly divided into a model group, a VitD₃ group, and an ESA group, with 15 rats in each group. In the ESA group, ESA was performed bilaterally at the "top-temporal anterior oblique line" with disperse-dense wave, a frequency of 2 Hz/100 Hz, and an intensity of 1 mA. Each session lasted for 30 min, once daily, for a total of 7 days. The VitD₃ group were treated with intragastric administration of 1,25-dihydroxyvitamin D₃ (1,25-VitD₃) solution (3 ng/100 g), once daily for 7 days. The neurological deficit scores and neurobehavioral scores were assessed before and after the intervention. After the intervention, the brain infarct volume was evaluated using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Immunofluorescence double staining was performed to detect the protein expression of CD32 and CD206 in the ischemic cortex. Western blot analysis was conducted to measure the protein expression of IL-6, IL-1β, and IL-10 in the ischemic cortex. RESULTS: Compared with the sham surgery group, the model group showed increased neurological deficit scores and neurobehavioral scores (P<0.01), increased brain infarct volume (P<0.01), increased protein expression of CD32, IL-6, and IL-1β in the ischemic cortex (P<0.01), and decreased protein expression of CD206 and IL-10 in the ischemic cortex (P<0.01). Compared with the model group, both the ESA group and the VitD₃ group showed decreased neurological deficit scores and neurobehavioral scores (P<0.01), reduced brain infarct volume (P<0.01), decreased protein expression of CD32, IL-6, and IL-1β in the ischemic cortex (P<0.01), and increased protein expression of CD206 and IL-10 in the ischemic cortex (P<0.01). Compared with the VitD₃ group, the ESA group had lower neurological deficit score (P<0.05), larger brain infarct volume (P< 0.05), and lower protein expression of CD32, CD206, IL-1β, and IL-10 in the ischemic cortex (P<0.01, P<0.05). CONCLUSION ESA could improve neurological function in MCAO rats, and its mechanism may be related to promoting microglial M1-to-M2 polarization and alleviating inflammatory damage.
Male ; Animals ; Rats ; Rats, Sprague-Dawley ; Ischemic Stroke ; Interleukin-10 ; Interleukin-6/genetics* ; Microglia ; Scalp ; Acupuncture Therapy ; Vitamins ; Infarction, Middle Cerebral Artery

OBJECTIVE To study the intervention effect and mechanism of Zhongfeng yure decoction on ischemic stroke model rats. METHODS Totally 85 rats were randomly divided into sham operation group （normal saline， n＝15）， model control group （normal saline， n＝18）， Nimodipine tablet group （positive control， 10.8 mg/kg， n＝18）， high-dose group of Zhongfeng yure decoction （20.52 g/kg， n＝17） and low-dose group of Zhongfeng yure decoction （5.13 g/kg， n＝17）， respectively. After 7 days of preventive continuous administration （once a day）， except for the sham operation group， the rats’ middle cerebral artery occlusion （MCAO） model was established by the modified suture method in other groups. After modeling， the rats in each group continued to be administered for 3 days. During experiment， general condition of the rats was observed， and the neurological function score was performed. After the last administration， the organ index was calculated， the cerebral infarction area and pathological changes of brain tissue were observed. The levels of tumor necrosis factor-α （TNF-α） and interleukin 6 （IL-6） in brain tissue and serum， and the average optical density value of caspase-3 and phosphorylated protein kinase B（p-AKT） protein in brain tissue were detected. RESULTS Three days after modeling， compared with sham operation group， the neurological function score， in brain tissue index， spleen tissue index， proportion of cerebral infarction area， the levels of TNF-α and IL-6 in brain tissue and serum， and the average optical density value of caspase-3 protein in brain tissue were significantly increased in the model control group （P＜0.05 or P＜0.01）； karyopyknosis， diffuse edema and other lesions appeared in brain tissue. Compared with the model control group， the above indexes in each administration group were improved to varying degrees. Among them， there were significant regression in brain tissue index， spleen tissue index， proportion of cerebral infarction area， TNF-α level in brain tissue and serum， and the average optical density values of caspase-3 protein and p-AKT protein in brain tissue of rats in high-dose group of Zhongfeng yure decoction （P＜0.05 or P＜0.01）. CONCLUSIONS Zhongfeng yure decoction has a certain intervention and therapeutic effect on MCAO model rats. The mechanism may be to reduce the secretion of inflammatory factors TNF-α and IL-6， down-regulate the expression of caspase-3 protein in ischemic brain tissue， up-regulate the expression of p-AKT protein， so as to protect the neurons.

Objective:To construct a whole-course and multidisciplinary nutrition management program for ovarian cancer patients undergoing chemotherapy.Methods:The related literatures published in the domestic and foreign journals were searched for drafting the first draft of the whole-course and multidisciplinary nutrition management program.Between May and July,2022,the Delphi method was used to conduct two rounds of expert consultations to 16 experts,and the analytic hierarchy process method was used to determine the weight of indicators at all levels.According to the experts'comments and suggestions,the draft was revised to formulate the final one.Results:In two rounds of Delphi expert consultations,expert positive coefficients were both 100％(16/16),and expert authority coefficients were both 0.933.The Kendall's coordination coefficients were 0.257 and 0.520,respectively(P＜0.001).Finally,6 first-level indicators,24 second-level indicators and 34 third-level indicators were determined in the final nutritional management program.Conclusions:Through literature research and semi-structured interviews,we develop a whole-course and multidisciplinary nutrition management program for ovarian cancer patients with chemotherapy.The program is reliable and clinically practical,and can be used for nutrition management of ovarian cancer patients in clinical practice.