In this study, the pharmacokinetic characteristics and tissue distribution of cannabidiol(CBD)/γ-polyglutamic acid-g-cholesterol(γ-PGA-g-CHOL) nanomicelles [CBD/(γ-PGA-g-CHOL)NMs] were investigated by pharmacokinetic experiments, and the effect of CBD/(γ-PGA-g-CHOL)NMs on the lipopolysaccharide(LPS)-induced inflammatory damage of cells was evaluated by cell experiments. CBD/(γ-PGA-g-CHOL)NMs were prepared by dialysis. The CBD concentrations in the plasma samples of male SD rats treated with CBD and CBD/(γ-PGA-g-CHOL)NMs were investigated, and the pharmacokinetic parameters were calculated and compared. UPLC-MS/MS was employed to determine the concentration of CBD in tissue samples. The heart, liver, spleen, lung, kidney, and muscle samples were collected at different time points to explore the tissue distribution of CBD and CBD/(γ-PGA-g-CHOL)NMs. The Caco-2 cell model of LPS-induced inflammation was established, and the cell viability, transepithelial electrical resistance(TEER), and secretion levels of inflammatory cytokines were determined to compare the anti-inflammatory activity between the two groups. The results showed that CBD/(γ-PGA-g-CHOL)NMs had the average particle size of(163.1±2.3)nm, drug loading of 8.78%±0.28%, and encapsulation rate of 84.46%±0.35%. Compared with CBD, CBD/(γ-PGA-g-CHOL)NMs showed increased peak concentration(C_(max)) and prolonged peak time(t_(max)) and mean residence time(MRT_(0-t)). Within 24 h, the tissue distribution concentration of CBD/(γ-PGA-g-CHOL)NMs was higher than that of CBD. In addition, both CBD and CBD/(γ-PGA-g-CHOL)NMs significantly enhanced Caco-2 cell viability and TEER, lowered the secretion levels of inflammatory cytokines, and alleviated inflammation. Moreover, CBD/(γ-PGA-g-CHOL)NMs demonstrated stronger anti-inflammatory effect. It can be inferred that γ-PGA-g-CHOL blank nanomicelles are good carriers of CBD, being capable of prolonging the circulation time of CBD in the blood, improving the bioavailability and tissue distribution concentration of CBD, and protecting against LPS-induced inflammatory injury. The findings can provide an experimental basis for the development and clinical application of oral CBD preparations.
Animals ; Cannabidiol/administration & dosage* ; Polyglutamic Acid/analogs & derivatives* ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Anti-Inflammatory Agents/administration & dosage* ; Micelles ; Caco-2 Cells ; Cholesterol/pharmacokinetics* ; Tissue Distribution ; Nanoparticles/chemistry*

OBJECTIVE: To identify the gene mutation types of 4 suspected β-thalassemia patients in Yunnan Province, and to analyze the genotypes and hematological phenotypes. METHODS: Whole genome sequencing was performed on the samples of 4 suspected β-thalassemia patients from the Dai ethnic group in a thalassemia endemic area of Yunnan Province, whose hematological phenotypes were not consistent with the results of common thalassemia gene mutations. The mutations of β-globin gene clusters were confirmed by polymerase chain reaction (PCR) and Sanger DNA sequencing technology. RESULTS: The 3.5 kb deletion in β-globin gene cluster (NC_000011.10: g. 5224302-5227791del3490bp) was detected in 4 patients' samples, of which 1 case was also detected with HbE mutation and 1 case with CD17 mutation. These 2 patients displayed moderate anemia phenotype, while the two patients with only the 3.5 kb deletion presented with other mild anemia phenotype. CONCLUSION Heterozygous carriers with rare 3.5 kb deletion of the β-globin gene cluster may develop mild anemia, compound mutations of the 3.5 kb deletion with other mutations may led to intermediate thalasemia with moderate to sever anemia. In areas with a high incidence of thalassemia, suspected patients should undergo genetic testing to avoid missing or misdiagnosing rare mutations.
Humans ; beta-Globins/genetics* ; Multigene Family ; beta-Thalassemia/genetics* ; Mutation ; Genotype ; Sequence Deletion ; Phenotype ; Male ; Female

A comprehensive phytochemical investigation of the leaves and twigs of Physalis angulata. var. villosa resulted in the isolation of 23 withanolide derivatives, including one novel 13,20-γ-lactone withanolide derivative (1) and three new withanolide derivatives (2-4). Architecturally, physalinin A (1) represents the first identified type B withanolide featuring a 13,20-γ-lactone moiety. The molecular structures of all isolates were elucidated using an integrated approach combining nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), infrared (IR) spectroscopy, and quantum chemical calculations to confirm structural assignments. The antiproliferative activities of all isolated withanolides were evaluated against four human cancer cell lines (HEL, HCT-116, Colo320DM, and MDA-MB-231). Among them, eight derivatives (2, 5-8, 14, 15, and 23) exhibited significant inhibitory effects, with half-maximal inhibitory concentration (IC₅₀) values of 0.18 ± 0.03 to 17.02 ± 0.21 μmol·L^-1. Structure-activity relationship (SAR) analysis suggested that the presence of an epoxide ring enhances anticancer activity, potentially through increased reactivity or specific interactions with molecular targets involved in cancer progression. These findings underscore the pharmacological potential of withanolides as promising lead compounds for the development of novel anticancer therapeutics.
Withanolides/isolation & purification* ; Physalis/chemistry* ; Humans ; Molecular Structure ; Cell Line, Tumor ; Antineoplastic Agents, Phytogenic/isolation & purification* ; Cell Proliferation/drug effects* ; Plant Leaves/chemistry* ; Plant Extracts/pharmacology*

AIM:To construct a recombinant adenovirus vector carrying the mouse ubiquitin-like with plant homeodomain and RING finger domains 1(Uhrf1)gene,validate the expression of Uhrf1 in neonatal mouse cardiomyo-cytes and explored its role in hydrogen peroxide(H2O2)-induced DNA damage.METHODS:The mouse Uhrf1 gene cod-ing sequence was amplified by polymerase chain reaction(PCR),digested,and inserted into the pADM-CMV-C-FH vec-tor to create the recombinant adenoviral plasmid ADM-Uhrf1.Following transfection into HEK293T cells,we generated re-combinant adenoviral particles,amplified,purified,and determined the titer.Neonatal mouse cardiomyocytes were infect-ed at an multiplicity of infection(MOI)of 50,UHRF1 protein expression was validated via Western blot and immunofluo-rescence staining.H2O2-induced DNA damage was explored along with adenovirus-mediated Uhrf1 overexpression to inves-tigate its role in DNA damage repair.RESULTS:ADM-Uhrf1 virus titer,determined by capsid immunofluorescence as-say,was 1.8×1013 pfu/L.Western blot confirmed a significant increase in UHRF1 protein expression(P＜0.05),with im-munofluorescence indicating predominant nuclear localization.Uhrf1 overexpression effectively inhibited the expression of the DNA damage marker,phosphorylated H2AX protein(γH2AX)(P＜0.01).CONCLUSION:We successfully con-structed a recombinant adenoviral vector carrying the mouse Uhrf1 gene,facilitating Uhrf1 overexpression in neonatal mouse cardiomyocytes.Furthermore,this overexpression effectively alleviated DNA damage in cardiomyocytes.

[Objective]To summarize the clinical experience of Professor CHEN Jian in treating periodic fever syndrome(PFS)in children from"latent fire".[Methods]Through outpatient follow-up study,collection of medical records and review of relevant literature,this paper summarized the etiology,pathogenesis and Professor CHEN's treatment experience of PFS in children from"latent fire",and a case was presented for verification.[Results]Professor CHEN believes that the core pathogenesis of PFS in children is"latent fire".Phlegm,poison and stasis are not only pathological products,but also important pathogenic factors throughout the course of the disease.Depression is the pathological state of the whole course of the disease.In the treatment,the method of supplementing Qi to remove heat or nourishing Yin to clear heat is used and combined with regulating Qi,and the self-prepared prescription is the effective treatment of PFS in children.The main symptom of the case was periodic fever,which was differentiated as Yin deficiency fever.The treatment of nourishing Yin and clearing heat,promoting Qi movement and relieving depression was applied,and good effect was achieved.[Conclusion]Professor CHEN's treatment of PFS in children from"latent fire"is effective,has certain guiding significance,and is worthy of clinical reference.

Objective:To investigate the role and mechanism of microRNA (miR)-4472 targeting PIN1 in regulating the nuclear factor kappa B (NF-κB)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in a rat model of hypoxic-ischemic encephalopathy (HIE).Methods:Between January 2022 and January 2024, sixty Sprague-Dawley rats were randomly assigned to six groups at The Second Hospital of Jiaxing using a random number table method: a normal control group, an HIE model group, an inhibition control group, a miR-4472 inhibition group, a miR-4472 inhibition + interference control group, and a miR-4472 inhibition + PIN1 interference group, with ten rats in each group. There was no significant difference in body mass among the six groups (all P > 0.05). Rat models of HIE were established using the Rice-Vannucci method. Behavioral performance was assessed using the Morris water maze test, while neurological function was evaluated using the Longa scoring method. Apoptosis was detected using the TUNEL assay, and the expression of NF-κB and STAT3 protein was measured using Western blot analysis. Results:Compared with the HIE model group, the miR-4472 inhibition group and the miR-4472 inhibition + interference control group showed a shortened escape latency, while the miR-4472 inhibition + PIN1 interference group exhibited an extended escape latency (all P < 0.05). The number of platform crossings in the miR-4472 inhibition + PIN1 interference group [(2.13 ± 0.54) times] was significantly lower than that in the HIE model group [(3.56 ± 0.71) times], the inhibition control group [(3.61 ± 0.87) times], the miR-4472 inhibition group [(5.47 ± 1.29) times], and the miR-4472 inhibition + interference control group [(5.58 ± 1.32) times] ( t = 5.07, 4.57, 7.55, 7.65, all P < 0.05). The Longa score in the miR-4472 inhibition + PIN1 interference group [(3.03 ± 0.30) points] was significantly lower than that in the HIE model group [(2.45 ± 0.54) points], the inhibition control group [(2.38 ± 0.69) points], the miR-4472 inhibition group [(1.27 ± 0.46) points], and the miR-4472 inhibition + interference control group [(1.29 ± 0.51) points] ( t = 2.97, 2.73, 10.13, 9.30, all P < 0.05). The apoptosis rate of hippocampal neurons in the miR-4472 inhibition + PIN1 interference group [(25.34 ± 6.16)%] was significantly lower than that in the HIE model group [(18.42 ± 5.46)%], the inhibition control group [(17.95 ± 4.38)%], the miR-4472 inhibition group [(8.89 ± 2.10)%], and the miR-4472 inhibition + interference control group [(9.13 ± 2.57)%] ( t = 2.97, 2.73, 10.13, 9.30, all P < 0.05). Compared with the HIE model group, the miR-4472 inhibition group and the miR-4472 inhibition + interference control group exhibited decreased gray values of NF-κB and STAT3 protein, while the miR-4472 inhibition + PIN1 interference group showed increased gray values of NF-κB and STAT3 protein (all P < 0.05). Conclusion:miR-4472 targets and regulates PIN1, which contributes to HIE injury through the activation of the NF-κB/STAT3 signaling pathway.

Objectives:Present study analyzed the efficacy and safety of percutaneous coronary intervention(PCI)using the distal transradial access(dTRA)for patients with complex coronary lesions. Methods:A total of 10 033 patients with complex coronary artery lesions(type B2 and type C lesions)who underwent percutaneous coronary intervention(PCI)via dTRA or conventional transradial access(TRA)at Fuwai Hospital between June 2021 and May 2022 were included(9 625 patients in the TRA group and 408 patients in the dTRA group).After propensity score matching,391 patients were included in each group.Baseline data,PCI intraoperative data(including lesion characteristics,intervention success rate,etc.),and incidence of major bleeding related to the access were compared between the two groups before and after propensity score matching. Results:Before propensity score matching,the proportions of patients with hypertension,hyperlipidemia,family history of coronary heart disease,history of myocardial infarction,and history of coronary artery bypass grafting were significantly higher in the dTRA group than in the TRA group(all P<0.05).After propensity score matching,the baseline data of the two groups were similar(all P>0.05).Before propensity score matching,compared with the TRA group,patients in the dTRA group had a higher proportion of patients with type B2 lesions,while the proportions of patients with type C lesions and those using intravascular ultrasound(IVUS)were lower(all P<0.05).The proportion of patients with chronic complete occlusion was similar between the two groups(P>0.05).After propensity score matching,compared with the TRA group,patients in the dTRA group had a lower proportion using IVUS and had a higher percent of stent implantation(both P<0.05).There was no statistically significant difference between the two groups in terms of SYNTAX score,guide catheter size,target lesion distribution,proportion of patients using intra-aortic balloon counterpulsation,success rate of intervention procedures,and incidence of major bleeding events related to the access(all P>0.05). Conclusions:Compared with the conventional TRA,interventional treatment of complex lesions through dTRA is equally safe and effective for patients with complex coronary lesions.

Objective:To investigate the central mechanism of moxibustion in treating chronic inflammatory visceral pain(CIVP)and its analgesic effect from the perspective of the p38 mitogen-activated protein kinase(MAPK)/Ets-like transcription factor 1(ELK1)signaling pathway in the spinal cord. Methods:Clean-grade male Sprague-Dawley rats were randomly divided into a normal group,a model group,a herb-partitioned moxibustion(HPM)group,a sham-HPM group,a p38 MAPK inhibitor group,and a dimethyl sulfoxide(DMSO)group.CIVP rat models were prepared using an enema mixture of 2,4,6-trinitrobenzene sulfonic acid solution and 50%ethanol.The HPM group was treated with HPM;the sham-HPM group was treated the same as the HPM group,but the moxa cones were not ignited;rats in the p38 MAPK inhibitor group received L5-L6 intrathecal injection of p38 MAPK inhibitor(SB203580);rats in the DMSO group received L5-L6 intrathecal injection of 2%DMSO.Abdominal withdrawal reflex(AWR),mechanical withdrawal threshold(MWT),and thermal withdrawal latency(TWL)were used to observe pain-related behaviors in each group.Hematoxylin-eosin staining was used to observe the morphological changes in rat colon tissue.Western blotting and real-time quantitative reverse-transcription polymerase chain reaction were used to detect the phosphorylated protein and mRNA expression of apoptosis signal-regulating kinase 1(ASK1),MAPK kinase(MKK)3/6,p38 MAPK,ELK1,and mitogen and stress-activated protein kinase 1(MSK1)in the spinal cord. Results:Compared with the normal group,CIVP rats had severe colonic inflammatory injuries,and the pathological injury scores increased significantly,along with increased AWR scores under different colorectal distension(CRD)stimulation pressures and decreased MWT and TWL;the mRNA and phosphorylated protein expression of p38 MAPK,ELK1,MSK1,ASK1,MKK3,and MKK6 all increased in the spinal cord(P<0.01).After HPM treatment,the colon injuries were repaired,and the pathological injury scores decreased;under different CRD stimulation pressures,the AWR scores decreased,and the MWT and TWL increased;the mRNA and phosphorylated protein expression of p38 MAPK,ELK1,ASK1,and MKK3 in the spinal cord also decreased,with statistically significant differences compared with the model group and the sham-HPM group(P<0.01).There were no significant differences in the above indicators between the HPM group and the p38 MAPK inhibitor group(P>0.05),and the same was true regarding the comparisons between the model group and the DMSO group. Conclusion:HPM exerted analgesic effects via downregulating the mRNA and phosphorylated protein expression of ASK1,MKK3,p38 MAPK,and ELK1 in the spinal cord of CIVP rats.The inhibition of spinal p38 MAPK/ELK1 signaling pathway activation may be one of the mechanisms by which HPM relieves pain in CIVP.

Objective:To discuss the clinical efficacy of treating lumbar muscle strain(LMS)with Tuina(Chinese therapeutic massage)plus Chinese medication hot compress. Methods:A total of 147 LMS patients were randomized into a Tuina group,a Chinese medication hot compress group,and a combined group,each consisting of 49 cases.The Tuina group received Tuina treatment;the Chinese medication hot compress group received Chinese medication hot compress treatment;and the combined group received the forementioned two therapies alternately.The three groups of patients were assessed using the visual analog scale(VAS)and Oswestry disability index(ODI)before treatment and after 2 weeks of treatment.A 2-month follow-up was also conducted to observe the relapse rate. Results:The VAS and ODI scores dropped significantly after treatment in all three groups compared with their baseline(P<0.05),and the combined group surpassed the other two groups in comparing the ODI score(P<0.05).The 2-month follow-up showed that the combined group had the lowest relapse rate among the three groups(P<0.05). Conclusion:Compared to each therapy used alone,Tuina plus Chinese medication hot compress can relieve pain,improve daily living function,and reduce the short-term relapse rate better in treating LMS patients.