Objective To prepare glucose transporter 1(Glut1)-targeted doxorubicin(DOX)-loaded liposomes(doxorubicin/polyphyllin H-liposomes,DOX/ppH-LPs)using polyphyllin H(ppH)instead of cholesterol as the liposomal membrane material,and to investigate their in vitro synergistic anti-tumor activity against non-small cell lung cancer(NSCLC).Methods DOX/ppH-LPs were prepared using thin-film hydration,and the formulation was optimized by single-factor investigation.The optimized DOX/ppH-LPs were characterized for morphology,particle size,polydispersity index(PDI),and zeta potential with transmission electron microscopy(TEM)and dynamic light scattering(DLS).Drug loading DL%was determined by high-performance liquid chromatography(HPLC).The storage stability was evaluated by observing in PBS at 4℃for 7 d,and the serum stability was observed in DMEM containing 10%fetal bovine serum(FBS)at 37℃for 48 h.In vitro drug release was studied in PBS at pH 7.4 and pH 5.0 values,respectively.Human NSCLC A549 cells were subjected as the model,MTT assay was performed to detect the proliferation inhibition by DOX/ppH-LPs at different concentrations(0.5,5.0,15.0 μg/mL)and the control group(ppH+DOX/LPs,a physical mixture of free ppH and DOX-loaded liposomes).Fluorescence microscopy was used to observe cellular uptake of DOX/ppH-LPs and DOX/LPs(containing 5 μg/mL DOX)at 15 min and 2 h.Live/dead cell staining was applied to assess apoptosis/necrosis induced by formulations(15 μg/mL DOX)after 48 h incubation.Transwell assay was conducted to evaluate inhibitory effect on cell migration and invasion,and the targeting property and in vitro synergistic anti-NSCLC activity of DOX/ppH-LPs were then comprehensively evaluated.Results The optimal formulation of DOX/ppH-LPs was determined as hydration temperature at 50℃,6 mg DOX,2 mg ppH,and 24 mg lecithin.The prepared DOX/ppH-LPs were in spherical shape,uniform distribution,and at an average particle size of 145.13±22.14 nm,a PDI of 0.15±0.05,a zeta potential of-23.92±1.73 mV,and a DL of 10.13±0.71%for DOX and(1.22±0.21)%for ppH.DOX/ppH-LPs maintained stable particle size,PDI,and exhibited significantly unchanged zeta potential after storage in PBS at 4℃for 7 d or incubation in DMEM containing 10%FBS at 37℃for 48 h,demonstrating excellent physical and serum stability.Both liposomes showed slow release at pH 7.4 value,while drug release was significantly accelerated at pH 5.0 value(P<0.05),indicating pH-sensitive release characteristics.MTT assay revealed that DOX/ppH-LPs exerted significantly stronger cytotoxicity against A549 cells than the ppH+DOX/LPs control group(P<0.05).Compared with ppH+DOX/LPs,DOX/ppH-LPs showed remarkably enhanced cellular uptake in A549 cells(P<0.05),with more DOX localized in the nucleus.Live/dead cell staining showed that at the same DOX concentration(15 μg/mL),the proportion of apoptotic/necrotic cells induced by DOX/ppH-LPs was significantly higher than that of the DOX/LPs control group.Transwell assay demonstrated that there were significantly less cells migrating and invading through the membrane in the DOX/ppH-LPs group than the ppH+DOX/LPs group.Conclusion Glut1-targeted doxorubicin-loaded liposomes(DOX/ppH-LPs)constructed by substituting cholesterol with ppH can target NSCLC cells,significantly enhance the in vitro synergistic anti-NSCLC activity of DOX and ppH.

Objective:To examine the impact of berberine on polycystic ovary syndrome (PCOS) in mice, and to investigate the effects of berberine on the intestinal flora and the intestinal flora on PCOS.Methods:A mouse model of PCOS was established by administering dehydroepiandrosterone in combination with high fat diet, and the mouse model was given a berberine treatment. The study consisted of a blank control group (C group), a PCOS model group (M group) and a berberine treatment group (T group). During the experiment, the mice were closely monitored through timed body weight measurements and estrous cycle monitoring; intraperitoneal glucose tolerance test and insulin tolerance test were done. Upon completion of the pharmacological intervention, the wet weights of liver, ovary and fat deposits of mice were assessed and subjected to HE staining to confirm the success of PCOS modeling and the efficacy of berberine. Additionally, fecal samples were analyzed for intestinal flora through 16S rRNA analysis.Results:The PCOS model was established successfully, berberine alleviated the disturbance of estrous cycle in mice, and significantly alleviated fat accumulation and metabolic abnormalities of glucose in mice. The cross-sectional area of fat pad cells in T group was (2 858±146) μm2, which was significantly lower than that in M group [(9 518±347) μm2], and the difference was statistically significant ( P<0.001). The blood glucose levels in T group were significantly lower than those in M group ( P<0.05). The composition and structure of intestinal flora in mice of M group with PCOS (compared with C group) and in mice of T group after berberine intervention (compared with M group) were significantly altered. However, alpha diversity did not change significantly among three groups ( P>0.05). Conclusion:Berberine could alleviate PCOS by intervening in the alterations of gut microbiota.

Objective To study the clinical and CT findings of bronchiolar adenoma. Methods Patients diagnosed with bronchiolar adenoma confirmed by surgical pathology at Linyi People's Hospital and Yantai Yuhuangding Hospital from 2016 to 2021 were collected. Their clinical and CT imaging features were retrospectively analyzed. Results Finally, 25 patients were collected, including 6 males and 19 females, aged 32-73 (58.6±10.1) years. The immunohistochemical Ki-67 (MIB1) of all lesions was <5%. The lesions were located in the upper and middle lobe of both lungs in 9 patients, lower lobes in 16 patients, extrapulmonary zone in 22 patients, intrapulmonary middle zone in 3 patients, round in 11 patients, irregular in 14 patients, well-defined in 22 patients, pure ground-glass/mixed ground-glass nodules in 6 patients, solid nodules in 19 patients. There were 11 patients with central small cavity, 18 patients with single bronchioles sign, 19 patients without adhesion with adjacent pleura, and 24 patients without mediastinal lymph node enlargement. Conclusion Bronchiolar adenomas usually occur in the middle-aged and elderly, mostly in the lower lobe of both lungs and the distribution of the peripheral lung field, most of the patients do not have any clinical symptoms, and the postoperative prognosis is good. CT may show large nodules or masses, pure ground-glass/mixed ground-glass nodules, irregular solid nodules and central small cavities. Irregular stellate nodules, central small cavity shadow, and single bronchiolar vascular bundle connected with the lesions are relatively specific imaging findings of bronchiolar adenoma.

Objective To observe the safety and efficacy of perioperative short-duration platelet membrane glycoprotein Ⅱ b/Ⅲ a receptor antagonist(GPI)in elderly patients undergoing primary percutaneous coronary intervention(PPCI)for acute ST-segment elevation myocardial infarction(STEMI)with high thrombotic burden.Methods A total of 140 elderly patients with acute STEMI admitted to our department from October 2021 to January 2024 were recruited and ran-domly divided into experimental group(short duration)and control group(standard duration),with 70 cases in each group.The occurrence of MACE,complete rate of ST segment resolution 2 h after surgery,blood flow classification thrombolysis in myocardial infarction(TIMI),left ventric-ular ejection fraction(LVEF),N-terminal pro-B-type natriuretic peptide(NT-proBNP),inhibition of platelet aggregation(IPA)by thromboelastography(TEG),maximum amplitude of adenosine diphosphate receptor pathway(MAADP),platelet-related indicators,and incidences of thrombocy-topenia and bleeding events were observed in the 2 groups after operation.Results There were no significant differences in postoperative recurrence of angina pectoris,acute heart failure,malignant arrhythmia,rate of ST segment resolution,TIMI blood flow grade 3,no reflow,LVEF,NT-proBNP level before discharge,and incidences of severe hemorrhage and thrombocytopenia during hospi-talization between the two groups(P＞0.05).The experimental group had significantly lower IPA[(76.1±15.3)％vs(96.3+19.4)％,P=0.016]and higher MAADP(45.6±8.2 mm vs 26.7±9.7 mm,P=0.028)in 18 h after operation,and lower incidence of minor bleeding(7.1％vs 24.3％,P＜0.05)during hospitalization when compared with the control group.Conclusion Under the premise of using a novel oral purinergic receptor P2Y12 inhibitor,short-term GPI is not inferior to the standard time course in achievement of IPA,with similar efficacy and relatively better safety.

Objective To construct curcumin pyrazole derivative by the reaction of diketone of curcumin and benzylhydrazine based on the above structure-activity relationship,and to explore its antioxidant activity to provide experimental basis for the development of curcumin antioxidant derivative.Methods Curcumin-N-substituted pyrazole derivative was synthesized from curcumin and benzylhydrazine.The structures of the derivative were confirmed by infrared spectroscopy(IR),nuclear magnetic resonance spectroscopy(1H-NMR,13C-NMR)and LC-MS.The antioxidant activity in vitro of the derivative was evaluated by determination of curcumin and its pyrazole derivative scavenging ability for 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid(ABTS)free radical.Results Curcumin pyrazole derivative was successfully synthesized.Curcumin and its pyrazole derivative showed good free radical scavenging effects in the range of 4.6-73.6,6.25-100 μg·mL-1,respectively,with a significant dose-effect relationship.The half-maximal inhibition(IC50)values of curcumin and its pyrazole derivatives determined by DPPH method were 14.24,40.37 μg·mL-1,respectively,while the IC50 values of curcumin and its pyrazole derivatives determined by ABTS method were 36.65,19.26 μg·mL-1,respectively.Conclusion The antioxidant activity of β-dione of curcumin was retained through the substitution of the pyrazole ring,and the curcumin pyrazole derivative deserves further investigation as a potential antioxidant.

Purpose To investigate the value of MYB and Notch1 immunohistochemical staining in the differential diagno-sis of classic adenoid cystic carcinoma of the breast(C-AdCC)and solid-basaloid adenoid cystic carcinoma of the breast(SB-AdCC).Methods MYB and Notch1 immunohistochemical staining were performed in 20 cases of C-AdCC,6 cases of SB-AdCC and 65 cases of other breast lesions in the archives of pa-thology department.26 cases of AdCC were detected by FISH,and 6 cases of SB-AdCC were detected by NGS.Results MYB immunohistochemical staining showed that C-AdCC(20/20)was moderately or strongly positive,while SB-AdCC(4/6)was mod-erately or strongly positive.Collagenous spherulosis(5/5)showed focal or diffuse weak positivity;Malignant adenomyoepi-thelioma(3/3)was focally moderately or strongly positive;8 matrix-producing carcinomas and 9 secretory carcinomas and 40 non-specific triple-negative breast cancers were negative.Immu-nohistochemistry of Notch1 showed diffuse moderate positive for SB-AdCC(3/6)and negative for C-AdCC(20/20).3 cases of malignant adenomyoepithelioma,5 cases of collagenous spherulo-sis,8 cases of matrix-producing carcinoma,9 cases of secretory carcinoma and 40 cases of non-specific triple-negative breast cancer were negative.FISH showed MYB gene disruption in C-AdCC(12/19)and NGS showed SB-AdCC(3/6)Notch1 muta-tion.Conclusion Moderately or strongly diffuse expression of MYB and Notch1 by immunohistochemistry can assist in the dif-ferentiation of C-AdCC from SB-AdCC,and it can be further clarified by molecular detection when it is difficult to distinguish malignant adenomyoepithelioma.

Objective To systematically review the efficacy of anti-inflammatory drugs in the treatment of diabetic nephropathy(DN).Methods Databases including PubMed,The Cochrane Library,EMbase,Web of Science,Scopus,Ovid,ProQuest,CBM,CNKI,WanFang Data,VIP and Duxiu data were electronically searched to collect randomized controlled trials(RCTs)of anti-inflammatory drugs for DN from inception to April 5,2022.Two reviewers independently screened the literature,extracted data,and assessed the risk of bias of the included studies.RevMan 5.4 software were then used to perform Meta-analysis.Results A total of 29 literature and 26 RCTs involving 4 095 patients were included.The results of Meta-analysis showed that compared with conventional treatment,conventional treatment combined with anti-inflammatory drugs could effectively reduce urinary albumin to creatinine ratio[SMD=-0.17,95％CI(-0.31,-0.03),P=0.02],urinary albumin excretion rate[SMD=-0.37,95％CI(-0.56,-0.18),P=0.000 1],urinary protein excretion rate[SMD=-0.97,95％CI(-1.29,-0.64),P=0.000 01],and glycosylated hemoglobin[SMD=-0.17,95％CI(-0.27,-0.08),P=0.000 4],while there was no significant difference in reducing serum creatinine[SMD=-0.04,95％CI(-0.19,0.1),P=0.57],urea nitrogen[MD=-0.23,95％CI(-0.50,0.04),P=0.09]and fasting blood glucose[SMD=-0.15,95％CI(-0.32,0.02),P=0.08].There was no statistically significant difference in changing glomerular filtration rate(GFR)[SMD=-0.04,95％CI(-0.15,0.07),P=0.47]with multiple drugs,except for a few drugs.Conclusion Conventional treatment combined with anti-inflammatory drugs can better improve the level of proteinuria in patients with DN,but the improvement of renal function is not obvious.Due to the limitations of the number of included studies and the duration of treatment,the above conclusion needs to be verified by more high-quality studies.

OBJECTIVE: Despite the combination of Scutellaria barbata D. Don and Scleromitrion diffusum (Willd.) R.J. Wang (SB-SD) being a recognized Chinese medicinal herbal pair that is commonly used in the treatment of ovarian cancer, there is a poor understanding of their pharmacological mechanisms. This study examines the antitumor properties and potential mechanisms of SB-SD on human ovarian cancer A2780 cells through a multi-omics approach, establishing a pharmacological basis for clinical utilization. METHODS: A range of mass ratios and reagents were used in the hot reflux extraction of SB-SD. The inhibitory effect of the SB-SD extracts on A2780 cell proliferation was assessed using the cell-counting kit 8 assay. A zebrafish tumor implantation model was used to evaluate the effects of SB-SD extracts on tumor growth and metastasis in vivo. Transcriptomics and proteomics were used to investigate alterations in biological pathways in A2780 cells after treatment with different concentrations of SB-SD extract. Cell cycle, cell apoptosis, intracellular free iron concentration, intracellular reactive oxygen species (ROS) concentration, malondialdehyde (MDA), and mitochondrial membrane potential were measured. Real-time quantitative reverse transcription polymerase chain reaction and Western blotting were utilized to investigate the effects of heme catabolism and ferritinophagy on ferroptosis induced by SB-SD extract in A2780 cells. RESULTS: The 70% ethanol extract of SB-SD (a mass ratio of 4:1) inhibited A2780 cell proliferation significantly with a half maximal inhibitory concentration of 660 μg/mL in a concentration- and time-dependent manner. Moreover, it effectively suppressed tumor growth and metastasis in a zebrafish tumor implantation model. SB-SD extract induced the accumulation of free iron, ROS, MDA, and mitochondrial damage in A2780 cells. The mechanisms might involve the upregulated expression of ferritinophagy-related genes microtubule-associated protein 1 light chain 3, autophagy-related gene 5, and nuclear receptor coactivator 4. CONCLUSION SB-SD extract effectively inhibited the development of ovarian cancer both in vitro and in vivo. Its mechanism of action involved inducing ferroptosis by facilitating heme catabolism and ferritinophagy. This herbal pair holds promise as a potential therapeutic option for ovarian cancer treatment and may be utilized in combination with routine treatment to improve the treatment outcomes of ovarian cancer patients. Please cite this article as: Wang Z, Liu M, Li GX, Zhang L, Ding KY, Li SQ, Gao BQ, Chen P, Choe HC, Xia LY, Yang YT, Liu Y, Sui X, Ma JN, Zhang L. A herbal pair of Scutellaria barbata D. Don and Scleromitrion diffusum (Willd.) R.J. Wang induced ferroptosis in ovarian cancer A2780 cells via inducing heme catabolism and ferritinophagy. J Integr Med. 2024; 22(6): 666-683.
Ferroptosis/drug effects* ; Female ; Humans ; Animals ; Scutellaria/chemistry* ; Ovarian Neoplasms/genetics* ; Zebrafish ; Cell Line, Tumor ; Ferritins/genetics* ; Plant Extracts/pharmacology* ; Heme/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Cell Proliferation/drug effects* ; Reactive Oxygen Species/metabolism* ; Antineoplastic Agents, Phytogenic/pharmacology* ; Autophagy/drug effects* ; Apoptosis/drug effects*

Objective The dense fibrous connective tissue that connects sub-occipital muscles which consist of the rectus capitis posterior minor muscle (RCPmi), rectus capitis posterior major muscle (RCPma), obliquus capitis inferior muscle (OCI) and nuchal ligament (NL) to the spinal dura mater (SDM), is described as the myodural bridge (MDB) in humans. The MDB is perceived as an essential anatomical structure and has been a subject of interest for clinicians. Studies have revealed that MDB may be related to the dynamic circulation of the cerebrospinal fluid (CSF) and a chronic cervicogenic headache. To date, the MDB is identified as a universal, existing structure in mammals and it exists in other vertebrates as well, such as Gallus domesticus and Rock pigeons in Avifauna, Siamese crocodile and Trachemys scripta elegans in Reptile. The current study is to further analyze different structures features of the MDB in sundry classes and provide the anatomical basis for functional studies. The JapaLura Splendida is the most common species in Lacertiformes, Reptilia. So we chose it as the experimental object to supply the morphological study of the MDB in Reptilia. Methods The study was based on gross anatomical dissection, thick sheet section, histological staining to observe the structural characteristics of the post-occipital area of twenty JapaLura Splendidas and the existence of the MDB. Results The deep post-occipital muscles were composed of the rectus capitis dorsal muscle (RCD) and the obliquus capital posterior (OCP) muscle. The RCD was merged by the rectus capitis dorsal major muscle (RCDma), the rectus capitis dorsal minor muscle (RCDmi) and the obliquus capitis anterior muscle (OCA). In the atlanto-occipital space, the dense fibrous bundles were found to originate from the ventral aspect of the RCD and run ventral, closely inserting into the SDM. In the atlanto-axial space, the dense fibrous bundles were found to originate from the ventral aspect of the OCP and run ventral, closely contacted with the SDM. These dense fibrous bundles were the collagen type I fibers with strong double refraction. Conclusion The result of this study indicates that the MDB is located between the post-occipital muscles and the SDM in JapaLura Splendida. The MDB of Japalura splendida may be related to the activities of the head and neck, and exert a physiological function similar to the MDB in humans.