OBJECTIVE: To investigate the Wnt signaling pathway and miRNAs mechanism of extracts of Plastrum Testudinis (PT) in the treatment of osteoporosis (OP). METHODS: Thirty female Sprague Dawley rats were randomly divided into 5 groups by random number table method, including sham group, ovariectomized group (OVX), ovariectomized groups treated with high-, medium-, and low-dose PT (160, 80, 40 mg/kg per day, respectively), with 6 rats in each group. Except for the sham group, the other rats underwent bilateral ovariectomy to simulate OP and received PT by oral gavage for 10 consecutive weeks. After treatment, bone mineral density was measured by dual-energy X-ray absorptiometry; bone microstructure was analyzed by micro-computed tomography and hematoxylin and eosin staining; and the expressions of osteogenic differentiation-related factors were detected by immunochemistry, Western blot, and quantitative polymerase chain reaction. In addition, Dickkopf-1 (Dkk-1) was used to inhibit the Wnt signaling pathway in bone marrow mesenchymal stem cells (BMSCs) and miRNA overexpression was used to evaluate the effect of miR-214 on the osteogenic differentiation of BMSCs. Subsequently, PT extract was used to rescue the effects of Dkk-1 and miR-214, and its impacts on the osteogenic differentiation-related factors of BMSCs were evaluated. RESULTS: PT-M and PT-L significantly reduced the weight gain in OVX rats (P<0.05). PT also regulated the bone mass and bone microarchitecture of the femur in OVX rats, and increased the expressions of bone formation-related factors including alkaline phosphatase, bone morphogenetic protein type 2, collagen type I alpha 1, and runt-related transcription factor 2 when compared with the OVX group (P<0.05 or P<0.01). Meanwhile, different doses of PT significantly rescued the inhibition of Wnt signaling pathway-related factors in OVX rats, and increased the mRNA or protein expressions of Wnt3a, β-catenin, glycogen synthase kinase-3β, and low-density lipoprotein receptor-related protein 5 (P<0.05 or P<0.01). PT stimulated the osteogenic differentiation of BMSCs inhibited by Dkk-1 and activated the Wnt signaling pathway. In addition, the expression of miR-214 was decreased in OVX rats (P<0.01), and it was negatively correlated with the osteogenic differentiation of BMSCs (P<0.01). MiR-214 mimic inhibited Wnt signaling pathway in BMSCs (P<0.05 or P<0.01). Conversely, PT effectively counteracted the effect of miR-214 mimic, thereby activating the Wnt signaling pathway and stimulating osteogenic differentiation in BMSCs (P<0.05 or P<0.01). CONCLUSION PT stimulates bone formation in OVX rats through β-catenin-mediated Wnt signaling pathway, which may be related to inhibiting miR-214 in BMSCs.
Animals ; MicroRNAs/genetics* ; Female ; Rats, Sprague-Dawley ; Wnt Signaling Pathway/genetics* ; Osteogenesis/genetics* ; Mesenchymal Stem Cells/cytology* ; Cell Differentiation/drug effects* ; Bone Density/drug effects* ; Ovariectomy ; Osteoporosis/drug therapy* ; beta Catenin/metabolism* ; Rats ; Intercellular Signaling Peptides and Proteins/metabolism* ; Drugs, Chinese Herbal/pharmacology*

The development of anticancer drugs to treat triple-negative breast cancer (TNBC) is an ongoing challenge. Immunogenic cell death (ICD) has garnered considerable interest worldwide as a promising synergistic modality for cancer chemoimmunotherapy. However, only few drugs or treatment modalities can trigger an ICD response and none of them exert a considerable clinical effect against TNBC. Therefore, new agents with potentially effective chemoimmunotherapeutic response are required. In this study, five new cyclometalated Ir(III) complexes containing isoquinoline alkaloid CˆN ligands were designed and synthesized. Among them, Ir-1 exhibited the highest in vitro cytotoxicity. Mechanistically, Ir-1 could trigger autophagy-dependent ferroptosis and a subsequent ferroptosis-dependent ICD response as well as indoleamine 2,3-dioxygenase (IDO) inhibition via reactive oxygen species (ROS)-mediated endoplasmic reticulum (ER) stress in MDA-MB-231 cells. When immunocompetent BALB/c mice were vaccinated with Ir-1-treated dying TNBC cells, antitumor CD8⁺ T-cell response and Foxp3⁺ T-cell depletion were induced, resulting in long-lasting antitumor immunity in TNBC cells. Moreover, combination therapy with Ir-1 and anti-PD1 could substantially augment in vivo therapeutic effects. Based on these results, Ir-1 is a promising candidate for chemoimmunotherapy against TNBC and its effects are mediated synergistically via ICD induction and IDO blockage.

6-Thioguanine(6-TG)is an antineoplastic agent used in treatment of acute leukemia.However,significant interindividual variability in dosing regimens and frequent clinical manifestations of hepatotoxicity and myelosuppression as adverse effects have affected its therapeutic efficacy.Consequently,the development of rapid analytical methods for 6-TG in clinical samples,enabling continuous therapeutic drug monitoring of plasma concentrations,holds substantial significance in optimizing dosage regimens,mitigating adverse reactions,and investigating drug metabolism mechanisms.In this study,multi-tipped gold nanostars(AuNSs)were prepared.With bis-(p-sulfonylphenyl)phenylphosphine molecule as the protecting agent and internal standard molecule,the AuNSs were assembled onto a highly sensitive surface-enhanced Raman(SERS)substrate for developing a ratio-based SERS quantitative analysis method for 6-TG in serum.The AuNSs containing multiple tips and gaps exhibited strong local surface plasmon resonance effect and SERS activity,ensuring the sensitivity of the analytical method.Furthermore,the introduction of internal standard molecules could improve the reproducibility,which guaranteed this method suitable for rapid analysis of drug molecules in complex samples.Quantitative analysis of 6-TG was achieved with linear detetion range of 1.0×10?4-1.0 mmol/L.In the spiked recovery experiments of serum,the RSD was less than 5.32%,and the recoveries were 94%-104%,which proved that this method could be used for rapid quantitative determination of 6-TG in serum.This method provided a powerful tool for studying drug pharmacokinetics,which could promote the optimization of the usage methods of anti-cancer drugs,and it was expected to further enhance the clinical efficacy and safety of 6-TG,enabling it to achieve the best therapeutic effect.

Objective:To investigate the current status of the implementation of home-based phototherapy (HBPT) in patients with vitiligo, and to analyze management needs among patients receiving HBPT.Methods:A questionnaire survey was conducted on the application of HBPT among patients with vitiligo who visited the outpatient clinic of the Hospital for Skin Diseases, Chinese Academy of Medical Sciences from December 2021 to November 2022. Additionally, the popularization and usage of HBPT as well as needs of patient management were investigated and analyzed in these vitiligo patients.Results:A total of 496 valid questionnaires were collected from 496 patients with vitiligo (241 males [48.5%] and 255 females [51.5%]) . Their ages at visit ranged from 2 to 67 (30.87 ± 12.36) years. The most commonly affected sites were the head and face (52.2%) , followed by hair (32.1%) , hands and feet (31.4%) , trunk (30.2%) , limbs (24.3%) , neck (19.5%) , and perineum (9.9%) . Among the participants, 320 (64.5%) were currently using or had used HBPT, and 352 (70.8%) expressed a willingness to learn more about HBPT usage guidelines and health education. Regarding the repigmentation outcomes after HBPT: among 312 patients, 54 (17.3%) reported complete recovery, 64 (20.5%) were markedly improved, 142 (45.5%) experienced improvement, and 52 (16.7%) showed no response. Adverse reactions occurred in 223 patients (71.5%) , of whom 28 (9.0%) experienced severe adverse reactions. The most desired guiding information was the adjustment method for phototherapy dosage and treatment duration (184/352, 52.3%) ; the most effective way to receive health education information was through verbal education by medical staff (177/352, 50.3%) .Conclusion:For vitiligo patients who were willing to accept and use HBPT, the most desired guiding information was the adjustment method for phototherapy dosage and treatment duration, and verbal health education by medical staff appeared to be the main way to obtain health education information.

Neuroblastoma(NB),the most common type of extracranial solid tumor in children,is char-acterized by high malignancy and poor prognosis,warranting in-depth investigation.In recent years,mi-croRNAs(miRNAs)have emerged as crucial post-transcriptional regulators playing pivotal roles in tu-morigenesis and progression.Building upon this background,the present study specifically focuses on in-vestigating miR-3910's biological functions and underlying molecular regulatory mechanisms in the NB SH-SY5Y cell line.Through bioinformatics analysis and transcriptome sequencing,we identified potential key target molecules of miR-3910,thereby providing genetic targets for the precise diagnosis and effective treatment of NB.In this study,qRT-PCR was employed to measure miR-3910 expression levels in SH-SY5Y cells transfected with mimic negative control and miR-3910 mimic.Compared to the nc group,miR-3910 expression was significantly upregulated in the mimic group(P＜0.01).The CCK-8 assay and scratch wound healing assay were used to quantitatively assess the impact of miR-3910 on cell prolif-eration and migration.Results showed that cell proliferation significantly increased at 48 h(P＜0.05),and migration ability was markedly enhanced at 48 h(P＜0.01).Flow cytometry was applied to deter-mine the effect of miR-3910 on cell cycle progression,revealing accelerated cell cycle progression,a re-duced proportion of G0/G,phase cells(P＜0.01),and a significant increase in S-phase cells(P＜0.05).Integrated bioinformatics analysis and high-throughput transcriptome sequencing predicted key molecular changes in SH-SY5Y cells following miR-3910 overexpression.Transcriptome sequencing and bioinformatics analysis identified six NB-related genes:EIF3CL(EIF3C),RNF103-CHMP3(VPS24),SULT1A4(SULT1A4),CORO7-PAM16(CORO7),H4C12(Histone H4),and TBC1D3(TBC1D3A/B/C)(aliases sourced from the GeneCards database).qRT-PCR and Western blotting(WB)results are consistency with sequencing results(P＜0.01).In conclusion,miR-3910 overexpression significantly promotes SH-SY5Y cell proliferation,migration,and cell cycle progression,while uncovering a series of potential key target molecules.These findings provide new insights into the pathogenesis of NB and offer a theoretical foundation and potential intervention targets for molecular-targeted therapy in NB.

Blood from a case of group B epidemic cerebrospinal meningitis identified in February 2024 in Ganzhou City,Jiangxi Province,and throat swabs from close contacts were collected for isolation and culture.The isolates were subjected to serogrouping,drug sensitivity testing,and whole genome sequencing and analysis,to provide a basis for epidemiological inves-tigation and clinical drug use.One strain of Neisseria meningitidis was isolated from the blood of the case and denoted group B.The MLST type was ST-7962,with no clonal group attribution.The phylogenetic tree showed that it was genetically close to the 1977 Shanghai carrier isolate(id-52231).Drug sensitivity results indicated that the strain was sensitive to 8 drugs:azithro-mycin,cefotaxime,minocycline,ceftriaxone,chloramphenicol,meropenem,rifampicin,and benzylpenicillin;resistant to cot-rimoxazole,levofloxacin,and ciprofloxacin;and showed an intermediate response to penicillin.This report describes the first case of ST-7962 group B meningoencephalitis found in Jiangxi Province.Monitoring of Neisseria meningitidis carriage,drug re-sistance,and molecular characteristics of strains in the healthy population in this region should be strengthened,to provide la-boratory support for the clinical use of medications,traceability,and control of the pathogen underlying meningoencephalitis infection.

Objective To investigate the role of Sneathia sanguinegens(S.sanguinegens)in the development of unexplained recurrent spontaneous abortion(URSA).Methods A case-control study was conducted to analyze the vaginal flora characteristics of 65 patients with URSA and 18 healthy controls through 16S rRNA gene sequencing.Toxicity profile of S.sanguinegens on human cervical cancer cells(ME-180),human umbilical vein endothelial cells(HUVEC)and human placental choriocarcinoma cells(JEG-3)was analyzed at the cellular level to assess the mechanism of it in adverse pregnancy outcomes.And S.sanguinegens was used to infect C57BL/6J mice to explore the toxic effect on living organisms.Results The relative abundance of Sneathia was increased in patients with URSA compared with healthy controls.It was positively correlated with the number of miscarriages,and was attributed to S.sanguinegens.We also found that S.sanguinegens damaged ME-180,JEG-3 and HUVEC cells.The degree of cellular damage was related to the level of S.sanguinegens added.Intravenous infection with S.sanguinegens caused inflammatory damage in several organs and extramedullary hematopoiesis in the spleen.Conclusion S.sanguinegens is closely related to URSA and should be emphasized in patients with high vaginal bacterial load.

To investigate body composition and associated factors in adolescents undergoing long-term regular sports training.

Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.

Objective To explore the impact of polymorphisms in the solute carrier organic anion transporter family member 1B1(SLCO1B1)and apolipoprotein E(APOE)genes on statins selection and prognosis in patients after percutaneous coronary intervention(PCI).Methods This retrospective study enrolled 612 patients with coronary heart disease hospitalized in Tianjin Beichen Hospital from January 2021 to February 2023.Among them,411 patients who underwent PCI were assigned to PCI group,and 201 patients who did not undergo PCI were assigned to non-PCI group.According to treatment methods,PCI group was further divided into two subgroups:observation subgroup(n=305)and control subgroup(n=106).Non-PCI group was also divided into two subgroups:observation subgroup(n=147)and control subgroup(n=54).Patients in all observation subgroups underwent the detection of SLCO1B1 and APOE gene polymorphisms before medication,and an initial individualized treatment plan(including the selection of appropriate statins and dosages)was formulated based on the detection results.Patients in control subgroups were given statins at conventional dosages.Basic data before medication,post-medication lipid compliance rates[lipid intention-to-treat(ITT)compliance rate and lipid per-protocol analysis(PP)compliance rate],time to lipid compliance,liver function indicators before and after medication,and prognosis,as well as SLCO1B1 and APOE genotyping and allele frequency distribution were compared between observation subgroup and control subgroup.Conditional logistic regression was used to analyze the relationship between different genotypes and prognostic susceptibility.Multifactor dimensionality reduction(MDR)was applied to analyze the interaction between SLCO1B1 genotype and APOE genotype.Results All genotypes of SLCO1B1 and APOE genes in observation subgroups conformed to the Hardy-Weinberg genetic equilibrium test(P>0.05).Compared with control subgroups,the observation subgroups had significantly higher lipid PP compliance rates after 6 and 12 months of treatment.After 3 months of treatment,the number of completed cases and compliant cases in observation subgroups was significantly higher,and the time to lipid compliance was significantly shorter than that in control subgroups(P<0.05).Compared with before treatment,there were no significant changes in alanine aminotransferase(ALT)and creatine kinase isoenzyme(CK-MB)levels in either observation subgroups or control subgroups(P>0.05);the levels of total cholesterol(TC),triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)were significantly decreased(P<0.001),while high-density lipoprotein cholesterol(HDL-C)level was significantly increased(P<0.01).After 12 months of treatment,the aspartate aminotransferase(AST)level in observation subgroups was significantly lower than that before medication and also lower than that in the corresponding control subgroups(P<0.05).Within PCI group:there were 1 case and 5 cases of myocardial infarction,2 cases and 6 cases of target vessel revascularization,0 cases and 3 cases of stroke,0 cases and 7 cases of recurrent angina pectoris,1 case and 5 cases of readmission in observation subgroup and control subgroup,respectively.There were statistically significant differences between observation subgroup and control subgroup(P<0.05).Within non-PCI group:there were 0 cases and 3 cases of cardiogenic death,2 cases and 6 cases of myocardial infarction,2 cases and 5 cases of target vessel revascularization,0 cases and 4 cases of stroke,0 cases and 8 cases of recurrent angina pectoris,and 2 cases and 4 cases of readmission in observation subgroup and control subgroups,respectively,with statistically significant differences between the two subgroups(P<0.05).In the recessive model,SLCO1B1 and APOE genes were significantly correlated with prognosis in both PCI observation subgroup and non-PCI observation subgroup(P<0.05).MDR analysis of the interaction between SLCO1B1 genotype and APOE genotype in PCI observation subgroup and non-PCI observation subgroup showed that there was a significant interaction between the genotypes,which was closely related to prognosis(P<0.05).Conclusion Individualized statin treatment regimens based on the detection of SLCO1B1 and APOE gene polymorphisms can improve the lipid compliance rate,shorten the time to lipid compliance,and improve the prognosis of patients after PCI.