Objective Changes in relevant indexes in the mouse model of early-onset ovarian insufficiency caused by Tripterygium wilfordii polyglycoside were analyzed,and the optimal time point for intervention was determined.Methods Forty female ICR mice were randomly divided into control and A,B,C,and D model groups with eight mice in each group.The control group was gavaged with purified water for 14 days(0.01 mL/10 g),and the remaining groups were administered a Tripterygium wilfordii polyglycoside suspension(80 mg/kg,0.01 mL/10 g)for 1 day(A model group),3 days(B model group),7 days(C model group),or 14 days(D model group),and samples were collected.Body weight and wet weights of the uterus and bilateral ovaries of mice were determined in each group.Serum FSH,LH,E2,P,AMH,INH-B,and T contents were measured using enzyme-linked immunoassays.HE staining was used to observe the number and developmental status of follicles and corpus luteum at all levels in mice of each group.TUNEL staining was used to detect the apoptosis in the ovaries of mice in each group.IHC detected expression of VEGFA,CD34,and EPO proteins in the ovaries of mice in each group.mRNA expression of HIF-1α,SDF-1,and CXCR4 in each group of mice was detected by PCR.Results Compared with the control group,changes in indicators in model A mice did not meet the POI modeling standard.The ovarian index,uterine index,and body weight of mice in the B model group were decreased significantly(P＜0.01),the weight of the C model group was decreased significantly(P＜0.01),and the ovarian index of the D model group was decreased significantly(P＜0.05).Serum contents of FSH and LH in B,C,and D model groups were increased(P＜0.05,P＜0.01),the E2,PROG,AMH,INH-B,and T contents were decreased(P＜0.01).The numbers of basal follicles,pre-sinus follicles,sinusoidal follicles,antral follicles,preovulatory follicles,and corpus luteum were decreased significantly(P＜0.05,P＜0.01)and the number of atresia follicles was increased significantly(P＜0.01)in B,C,and D model groups.The apoptotic area of TUNEL staining in A,B,C,and D model groups was increased significantly(P＜0.05,P＜0.01).Expression of CD34,VEGFA,and EPO in B,C,and D model groups was decreased significantly(P＜0.05,P＜0.01).mRNA expression of HIF-1α,SDF-1,and CXCR4 in A and B model groups was significantly increased(P＜0.05,P＜0.01).Compared with the B model group,the relevant indexes of C and D model groups were changed significantly,indicating that C and D models were more serious and tended to develop POF.Conclusions The B model group is the turning point of ovarian function from impaired POI to irreversible POF,suggesting that 3 days of administrating Tripterygium wilfordii polyglycoside is optimal to induce a POI disease model for effective drug intervention.

OBJECTIVE: To study the in vitro and in vivo antitumor effects of the polysaccharide of Alocasia cucullata (PAC) and the underlying mechanism. METHODS: B16F10 and 4T1 cells were cultured with PAC of 40 µg/mL, and PAC was withdrawn after 40 days of administration. The cell viability was detected by cell counting kit-8. The expression of Bcl-2 and Caspase-3 proteins were detected by Western blot and the expressions of ERK1/2 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). A mouse melanoma model was established to study the effect of PAC during long-time administration. Mice were divided into 3 treatment groups: control group treated with saline water, positive control group (LNT group) treated with lentinan at 100 mg/(kg·d), and PAC group treated with PAC at 120 mg/(kg·d). The pathological changes of tumor tissues were observed by hematoxylin-eosin staining. The apoptosis of tumor tissues was detected by TUNEL staining. Bcl-2 and Caspase-3 protein expressions were detected by immunohistochemistry, and the expressions of ERK1/2, JNK1 and p38 mRNA were detected by qRT-PCR. RESULTS: In vitro, no strong inhibitory effects of PAC were found in various tumor cells after 48 or 72 h of administration. Interestingly however, after 40 days of cultivation under PAC, an inhibitory effect on B16F10 cells was found. Correspondingly, the long-time administration of PAC led to downregulation of Bcl-2 protein (P<0.05), up-regulation of Caspase-3 protein (P<0.05) and ERK1 mRNA (P<0.05) in B16F10 cells. The above results were verified by in vivo experiments. In addition, viability of B16F10 cells under long-time administration culture in vitro decreased after drug withdrawal, and similar results were also observed in 4T1 cells. CONCLUSIONS Long-time administration of PAC can significantly inhibit viability and promote apoptosis of tumor cells, and had obvious antitumor effect in tumor-bearing mice.
Mice ; Animals ; Alocasia/metabolism* ; MAP Kinase Signaling System ; Caspase 3/metabolism* ; Apoptosis ; RNA, Messenger/metabolism*

The bacterial ATP-competitive GyrB/ParE subunits of type II topoisomerase are important anti-bacterial targets to treat super drug-resistant bacterial infections. Herein we discovered novel pyrrolamide-type GyrB/ParE inhibitors based on the structural modifications of the candidate AZD5099 that was withdrawn from the clinical trials due to safety liabilities such as mitochondrial toxicity. The hydroxyisopropyl pyridazine compound 28 had a significant inhibitory effect on Gyrase (GyrB, IC₅₀ = 49 nmol/L) and a modest inhibitory effect on Topo IV (ParE, IC₅₀ = 1.513 μmol/L) of Staphylococcus aureus. It also had significant antibacterial activities on susceptible and resistant Gram-positive bacteria with a minimum inhibitory concentration (MIC) of less than 0.03 μg/mL, which showed a time-dependent bactericidal effect and low frequencies of spontaneous resistance against S. aureus. Compound 28 had better protective effects than the positive control drugs such as DS-2969 ( 5) and AZD5099 ( 6) in mouse models of sepsis induced by methicillin-resistant Staphylococcus aureus (MRSA) infection. It also showed better bactericidal activities than clinically used vancomycin in the mouse thigh MRSA infection models. Moreover, compound 28 has much lower mitochondrial toxicity than AZD5099 ( 6) as well as excellent therapeutic indexes and pharmacokinetic properties. At present, compound 28 has been evaluated as a pre-clinical drug candidate for the treatment of drug-resistant Gram-positive bacterial infection. On the other hand, compound 28 also has good inhibitory activities against stubborn Gram-negative bacteria such as Escherichia coli (MIC = 1 μg/mL), which is comparable with the most potent pyrrolamide-type GyrB/ParE inhibitors reported recently. In addition, the structure-activity relationships of the compounds were also studied.

Objective To explore the diagnostic efficacy of American Thyroid Association(ATA)guidelines,American College of Radiology Thyroid Imaging Report and Data System(ACR-TIRADS),and Chinese Thyroid Imaging Reporting and Data System(C-TIRADS)alone and combined with BRAF^V600E mutation in atypia of undetermined significance/follicular lesion of undetermined significance(AUS/FLUS).Methods A total of 138 patients who underwent ultrasound-guided fine needle aspiration(FNA)in the Chinese PLA General Hospital from January 2020 to May 2023 were selected.The clinicopathological and ultrasound characteristics were retrospectively analyzed for each nodule.Each nodule underwent preoperative BRAF^V600E mutation testing and was diagnosed according to the ATA guidelines,ACR-TIRADS,and C-TIRADS.The diagnostic efficacy of ATA guidelines,ACR-TIRADS,and C-TIRADS alone and combined with BRAF^V600E mutation was assessed based on the results of histopathological diagnosis.Results The 138 AUS/FLUS thyroid nodules included 45(32.6%)benign ones and 93(67.4%)malignant ones.The patient age(t=1.444,P=0.151),gender(χ²=0.259,P=0.611),and location of nodules(χ²=2.055,P=0.358)had no statistical significance for the differentiation between benign and malignant nodules,while nodule size(Z=2.500,P=0.012),echo(χ²=14.693,P<0.001),composition(χ²=17.075,P<0.001),aspect ratio ≥1(χ²=9.477,P=0.002),and microcalcification(χ²=6.892,P=0.009)were of significance for the differentiation.When applied alone,BRAF^V600E mutation showed high specificity(95.56%)and positive predictive value(95.65%).Among the three ultrasound grading systems,ACR-TIRADS had the highest sensitivity(χ²=37.923,P<0.001;χ²=40.462,P<0.001)and accuracy(χ²=81.595,P<0.001;χ²=76.912,P<0.001),while C-TIRADS had the highest specificity(χ²=11.746,P<0.001;χ²=21.235,P<0.001).However,the three systems showed no statistically significant difference in the diagnostic efficiency when applied alone(Z=1.177,P=0.239;Z=0.213,P=0.831;Z=1.016,P=0.310).The combination of BRAF^V600E mutation with ACR-TIRADS or C-TIRADS improved the diagnostic efficacy of BRAF^V600E mutation in distinguishing the benign and malignant AUS/FLUS nodules(Z=2.107,P=0.035;Z=2.752,P=0.006).The combination of ATA guidelines with BRAF^V600E mutation increased the diagnostic accuracy of BRAF^V600E mutation(χ²=20.679,P<0.001),while it had no statistically significant difference in distinguishing the benign and malignant AUS/FLUS nodules(Z=1.321,P=0.186).The combination of ATA guidelines,ACR-TIRADS,or C-TIRADS with BRAF^V600E mutation improved the diagnostic efficacy of ultrasound grading systems for AUS/FLUS nodules(Z=2.770,P=0.006;Z=2.770,P=0.006;Z=2.890,P=0.004).Specifically,ACR-TIRADS combined with BRAF^V600E mutation showed the highest sensitivity(χ²=4.712,P=0.030;χ²=4.712,P=0.030),while C-TIRADS combined with BRAF^V600E mutation showed the highest accuracy(χ²=77.627,P<0.001;χ²=85.827,P<0.001).However,there were no statistically significant differences in diagnostic performance between the combinations(Z=1.276,P=0.202;Z=0.808,P=0.419;Z=1.615,P=0.106).Conclusion ATA guidelines,ACR-TIRADS,and C-TIRADS combined with BRAF^V600E mutation can improve the diagnostic efficacy of BRAF^V600E mutation or ultrasound grading system alone in AUS/FLUS nodules,which can facilitate the further management and treatment of such patients.
Humans ; United States ; Infant ; Thyroid Neoplasms/genetics* ; Proto-Oncogene Proteins B-raf/genetics* ; Adenocarcinoma, Follicular/pathology* ; Retrospective Studies ; Data Systems ; Thyroid Nodule/genetics* ; Ultrasonography/methods* ; Mutation ; China ; Radiology

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.

Abstract: Viral shedding of SARS-CoV-2 is a continuous dynamic process, which can be divided into latent stage, initial stage, peak stage and decreasing stage according to the characteristics of viral shedding. After being infected with SARS-CoV-2, the infected person generally stays in the latent period for 1-3 days, which is characterized by continuous negative nucleic acid test results and no infectiousness, and the risk of infection for close contacts is very low. At the initial stage of viral shedding is characterized by a rapid decline in the Ct value of nucleic acid tests in a short time, and clinical symptoms gradually appear. The infectiousness of the infected person gradually increases during this period, and the risk of infection for close contacts also gradually increases, but it is still in the early stage of infection, the possibility of viral shedding is low, and the risk of infection of secondary close contacts is low. The peak of viral shedding is characterized by low Ct value in nucleic acid test and obvious clinical symptoms; during this period, the infected person is the most infectious, and the risk of infection of the contact is the highest, so the scope of close contacts should be expanded appropriately. The decreasing period is characterized by the gradual increase of Ct value of nucleic acid test and the gradual disappearance of clinical symptoms; during this period, the infectiousness of the infected person gradually decreases to disappear. In an outbreak, an infected person in the decreasing phase is more likely to be an early infected person in the transmission chain. If infected individuals in the decreasing phase are found in an area without a SARS-CoV-2 epidemic, it suggests that the local outbreak epidemic has been spreading for some time and may be larger in scale. According to the characteristics of viral shedding, risk personnel can be determined more scientifically and accurately, so as to minimize the risk and reduce the waste of epidemic prevention resources.

Objective To evaluate the efficacy of ultrasound and computed tomography (CT) in diagnosing cervical lymph node metastasis (CLNM) of papillary thyroid carcinoma (PTC). Methods The patients with PTC treated by surgery in the Chinese PLA General Hospital from January 2016 to January 2021 were selected for analysis.All the patients underwent preoperative ultrasound and CT examinations,the diagnostic values of which for CLNM were retrospectively analyzed. Results A total of 322 PTC patients were enrolled in this study,including 242 with CLNM and 80 with non-CLNM.The CLNM group and non-CLNM group had significant differences in age,tumor size,and maximum size of lateral CLNM (χ²=20.34,27.34,and 4.30,respectively,all P<0.001).For the central compartment,lateral compartment,and overall compartment,ultrasound diagnosis showed higher sensitivity (χ ²=82.26,P<0.001;χ²=114.01,P<0.001;χ²=82.26,P<0.001) and accuracy (χ²=20.27,P<0.001;χ²=15.56,P<0.001;χ²=44.00,P<0.001) than CT,and had no significant differences from ultrasound combined with CT (all P>0.05).However,ultrasound diagnosis had lower specificity than CT (χ²=17.01,P<0.001;χ²=21.29,P<0.001) in the central compartment and lateral compartment.Receiver operating characteristic curve analysis showed that in the central compartment,lateral compartment,and overall compartment,ultrasound diagnosis had larger AUC than CT (Z=2.99,P=0.003;Z=3.86,P<0.001;Z=4.47,P<0.001) and had no significant difference from ultrasound combined with CT (Z=1.87,P=0.062;Z=1.68,P=0.093;Z=1.61,P=0.107). Conclusions Ultrasound and CT have their own advantages in the diagnosis of central and lateral CLNM.In general,ultrasound has better performance than CT in the diagnosis of CLNM.
Humans ; Lymphatic Metastasis/diagnostic imaging* ; Retrospective Studies ; Thyroid Cancer, Papillary/pathology* ; Thyroid Neoplasms/surgery* ; Tomography, X-Ray Computed ; Ultrasonography/methods*

ObjectiveThis study aims to explore the potential molecular mechanism of Gegen Qinliantang （GQL） in the intervention of atherosclerosis （AS） based on network pharmacology and molecular docking. MethodThe active components and targets of each medicinal in GQL were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and AS-related genes from 7 databases. Thereby， the anti-AS targets of GQL were screened out. Cytoscape 3.8.0 was employed to construct the "component-target" network， and STRING the protein-protein interaction （PPI） network. Core targets were screened out with CytoNCA. R clusterProfiler was used for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of target genes， which were then visualized. Finally， molecular docking of the top ten active components with the core targets of AS was performed and the binding affinity was compared with that between atorvastatin and the core targets. ResultIn the end， 150 active components of GQL， 20 289 AS targets， and 213 common targets were retrieved， and 48 core common targets were screened out. They were mainly involved in the GO terms of nuclear receptor activity， ligand activation， and transcription factor activity and the pathways of fluid shear force and AS， advanced glycation end products-receptor for advanced glycation end products （AGE/RAGE）， interleukin-17 （IL-17）， tumor necrosis factor （TNF）， Toll-like receptor pathways and other signaling pathways closely related to AS. The molecular docking results showed that the effective components of GQL had high binding affinity to core targets of AS， and the binding affinity was even higher than that between the atorvastatin and core targets. The five groups with high binding affinity were puerarin-TNF， baicalein-inducible nitric oxide synthase 2 （NOS2）， puerarin-NOS2， and formononetin-NOS2， wogonin-NOS2. ConclusionThe above result provides new ideas for further exploration of this classical decoction.

Despite being a common therapy for hepatocellular carcinoma (HCC), insufficient thermal ablation can leave behind tumor residues that can cause recurrence. This is believed to augment M2 inflammatory macrophages that usually play a pro-tumorigenic role. To address this problem, we designed d-mannose-chelated iron oxide nanoparticles (man-IONPs) to polarize M2-like macrophages into the antitumor M1 phenotype. In vitro and in vivo experiments demonstrated that man-IONPs specifically targeted M2-like macrophages and accumulated in peri-ablation zones after macrophage infiltration was augmented under insufficient microwave ablation (MWA). The nanoparticles simultaneously induced polarization of pro-tumorigenic M2 macrophages into antitumor M1 phenotypes, enabling the transformation of the immunosuppressive microenvironment into an immunoactivating one. Post-MWA macrophage polarization exerted robust inhibitory effects on HCC progression in a well-established orthotopic liver cancer mouse model. Thus, combining thermal ablation with man-IONPs can salvage residual tumors after insufficient MWA. These results have strong potential for clinical translation.

Objective To discuss the value of contrast-enhanced ultrasound(CEUS)parameters in evaluating the formation of Kimmelstiel-Wilson(K-W)nodules in diabetic nephropathy(DN).Methods Sixty-two patients pathologically diagnosed with DN and undergoing CEUS in the First Medical Center of Chinese PLA General Hospital from March 2017 to January 2020 were assigned into two groups according to whether K-W nodules were formed.The cortical CEUS parameters and the ratios of cortical to medullary CEUS parameters were compared between the two groups.Results The 62 patients included 19 patients without K-W nodules(group A)and 43 patients with K-W nodules(group B).The median rise time(
Contrast Media ; Diabetes Mellitus ; Diabetic Nephropathies/diagnostic imaging* ; Humans ; Ultrasonography