The persistent high prevalence and poor survival outcomes of lung cancer underscore the urgent need for innovative therapeutic modalities. Here, we present a novel multifunctional delivery platform for the synergistic treatment of lung malignancies, combining in situ-triggerable photodynamic therapy (PDT) with radiotherapy. The new platform CLL was developed by loading a new reactive oxygen species (ROS)-triggerable photosensitizer, luminol-conjugated chlorin e6 (Ce6), into liposomes. CLL can be activated through the bioluminescence resonance energy transfer effect under oxidative stress, thereby producing singlet oxygen for targeted tumor treatment without external irradiation. In vitro studies showed significant cytotoxic effects of CLL in both 4T1 and A549 tumor cells. Furthermore, a PDT-radiopharmaceutical combination nanotherapy CLL-¹⁷⁷Lu was engineered by incorporating the radionuclide ¹⁷⁷Lu into CLL. CLL-¹⁷⁷Lu demonstrated synergistic antitumor effects in 4T1 and A549 tumor cells, as well as in mouse models of 4T1 breast cancer lung metastasis or A549 tumor xenografts. Mechanistically, CLL-¹⁷⁷Lu can induce singlet oxygen/ROS generation, enhance tumor cell apoptosis, and promote M1 macrophage-mediated immunotherapy. Preliminary assessments showed a favorable profile for CLL-¹⁷⁷Lu, highlighting its potential as a promising nanotherapy for cancer treatment. Additionally, CLL can serve as a versatile platform for delivering a range of therapies to achieve synergistic antitumor effects.

Accumulating evidence has demonstrated that nucleic acid-based therapies are promising for atherosclerosis. However, nearly all nucleic acid delivery systems developed for atherosclerosis necessitate injection, which results in rapid elimination and poor patient compliance. Consequently, oral delivery strategies capable of targeting atherosclerotic plaques are imperative for nucleic acid therapeutics. Herein we report the development of yeast-derived capsules (YCs) packaging an antisense oligonucleotide (AM33) targeting microRNA-33 (miR-33) for the oral treatment of atherosclerosis. YCs provide stability for AM33, preventing its premature release in the gastrointestinal tract. AM33-containing YCs, defined as YAM33, showed high transfection in macrophages, thus promoting cholesterol efflux and inhibiting foam cell formation by regulating the target genes/proteins of miR-33. Orally delivered YAM33 effectively accumulated within atherosclerotic plaques in ApoE ^-/- mice, primarily by transepithelial absorption via M cells in Peyer's patches and subsequent translocation via macrophages through the lymphatic system. Inhibition of miR-33 by oral YAM33 significantly delayed the progression of atherosclerosis. Moreover, oral treatment with YCs co-delivering AM33 and atorvastatin afforded significantly enhanced anti-atherosclerotic effects. Our findings suggest that yeast-based microcapsules represent an effective carrier for oral delivery of nucleic acids, either alone or in combination with existing drugs, offering a promising approach for precision therapy of atherosclerotic diseases.

Objective:To construct a novel chimeric antigen receptor T (CAR-T) cell targeting CD138 and to investigate its cytotoxicity against myeloma cells.Methods:The hybridoma strain that can stably secrete the CD138 monoclonal antibody (mAb) was prepared and obtained through monoclonal antibody screening technology. The hybridoma strain cells were intraperitoneally injected into mice to produce ascites containing monoclonal antibodies, which were then collected and purified to obtain pure CD138 mAb. Further examinations were performed to assess the biological characteristics of CD138 mAb. The variable region sequence of this antibody was amplified through reverse transcription polymerase chain reaction and was used as the antigen recognition domain of CD138 CAR, which was subsequently expressed on the surface of T cells by lentiviral infection. Flow cytometry was employed to assess the phenotype of CD138 CAR-T cells. In vitro cytotoxicity and degranulation assays were performed to evaluate their antitumor effects.Results:① We successfully prepared anti-human CD138 antibody hybridoma cell lines and screened a hybridoma cell strain, 5G2, which could persistently and stably secrete the anti-CD138 antibody. ② The purified CD138 (5G2) mAb can especially recognize CD138 + cells with a binding affinity constant (K D) of 6.011×10 -9 mol/L and showed no significant binding activity with CD138 - cells. ③The variable region sequence of the CD138 (5G2) antibody was obtained using molecular cloning technology, and CD138 (5G2) CAR was successfully constructed and expressed on T cells through lentivirus infection and, concurrently, demonstrated effective binding to recombinant human CD138 protein.④ The proliferation of T cells transduced with the CD138 (5G2) CAR was highly efficient. The phenotype analysis revealed that CD138 (5G2) CAR-T cells exhibited a greater tendency to differentiate into central memory T cells and memory stem T cells, with a reduced proportion of terminally differentiated effector memory subsets. ⑤CD138 (5G2) CAR-T cells demonstrated specific cytotoxicity against CD138 + myeloma cell line H929, whereas CD138 - cell line K562 remained unaffected. The percentage of residual H929 cells was (12.92±8.02) % after co-culturing with CD138 (5G2) CAR-T cells, while (54.25±15.79) % was left in the Vector-T group (E∶T=1∶2; P<0.001). ⑥Results of degranulation assays demonstrated a significant activation of CD138 (5G2) CAR-T cells after co-culture with the H929 cell line, whereas no significant activation was observed in Vector-T cells [ (25.78±3.35) % vs (6.13±1.30) %, P<0.001]. ⑦After co-culturing with CD138 + cells, CD138 (5G2) CAR-T cells exhibited a significant increase in cytokine secretion compared to the Vector-T group [interleukin-2: (1 697.52±599.05) pg/ml vs (5.07±1.17) pg/ml, P<0.001; interferon-γ: (3 312.20±486.38) pg/ml vs (9.28±1.46) pg/ml, P<0.001; and tumor necrosis factor-α: (1 837.43±640.49) pg/ml vs (8.75±1.65) pg/ml, P<0.001]. However, no significant difference was observed in cytokine secretion levels between the two groups after co-culturing with CD138 - cells. Conclusion:This study successfully prepared a novel monoclonal antibody against CD138, and CAR-T cells constructed with the antigen recognition domain derived from this 5G2 mAb demonstrated effective antitumor activity against myeloma cells. This can be used as a new option for the detection of the CD138 antigen and proposes a novel strategy for multiple myeloma immunotherapy.

Objective To construct vitamin E(tocopherol,VE)-based liposomes with both hydrogen peroxide-scavenging function and anticoagulant effect,and explore whether they can be used to treat acute ischemic stroke and then evaluate their in vitro anticoagulant effect.Methods Two molecules of VE were chemically bonded through peroxalate bond to obtain oxalate vitamin E(OVE).The anticoagulant unit LA-Hirudin was obtained by amidation reaction by connecting the anticoagulant drug hirudin to linoleic acid(LA).The structures of the synthesized products were characterized by using H nuclear magnetic resonance(1HNMR)and FT-IR spectra.Hirudin-Oxalic acid divitamin E ester liposome(HOL)was prepared by thin film dispersion and characterized with dynamic light scattering(DLS)and transmission electron microscopy(TEM).Mouse brain-derived endothelial cells(bEnd3)and human umbilical vein endothelial cells(HUVEC)were applied as the object of cell experiments.The levels of H2O2,and cellular inflammatory factors TNF-α,IL-6 and IL-1 β were determined by ELISA to evaluate the effect of the liposomes on the severity of cellular inflammation.Cell counting kit-8(CCK-8)assay and cell proliferation assay(5-Ethynyl-2'-deoxyuridine,Edu)were used to detect the effects of the liposomes on cell count and proliferation.The anticoagulant effect of the prepared liposomes was detected by using whole blood of mice.Results 1HNMR spectrum showed that OVE was successfully synthesized,and the FT-IR spectrum displayed that hirudin and linoleic acid were successfully bonded to obtain LA-Hirudin.The results of DLS and TEM indicated that the prepared liposomes were spheroids,in uniform distribution,with an average particle size of 161 nm.The cell experiments showed that the liposomes significantly inhibited the secretion of H2O2 and inflammatory factors,and exerted obvious antioxidant and anti-inflammatory effects in the model cells(P＜0.05).CCK-8 assay and Edu proliferation assay proved that the liposomes suppressed the decreased cell count and alleviated the inhibited cell proliferation caused by LPS(P＜0.01).Anticoagulation experiment revealed that the liposomes had significant inhibitory effect of CaCl2-induced coagulation reaction(P＜0.01).Conclusion A hydrogen peroxide-scavenging function material,OVE is synthesized,and then anticoagulant drug hirudin and OVE are used to prepare OVE thrombolytic liposomes,HOL.The liposomes have antioxidant and anti-inflammatory effects at the cellular level and also have significant anticoagulant effect.

Objective:To summarize the clinical features of epilepsy and (or) developmental delay associated with KCNB1 gene variants in children.Methods:A case series study was conducted on 24 children with KCNB1 gene variants associated with epilepsy and (or) developmental delay who were treated at the Children′s Medical Center of Peking University First Hospital and the Department of Neurology of Shenzhen Children′s Hospital from July 2015 to June 2024. The manifestations of seizures, electroencephalogram (EEG) and genetic test results of those children were analyzed.Results:All the KCNB1 gene variants were de novo, involving 20 different variation, including 15 missense variations, 3 frameshift variations and 2 nonsense variations. There were 7 novel variations. Among the 24 developmental and epileptic encephalopathy children, there were 14 boys and 10 girls. The last follow-up age ranged from 9 months to 13 years and 9 months. Seizures were present in 21 children (88%), with onset ranging from 1 month to 7 years, and 76% (16/21) began before 2 years of age. The seizure types included focal seizures in 15 children (71%), epileptic spasms, myoclonic seizures, and generalized tonic-clonic seizures in 6 children respectively, atypical absence seizures in 4 children, and myoclonic atonic seizures in 1 child. Seventeen children (81%) had a cluster of seizures and 5 had a history of focal status epilepticus with impaired consciousness. All 24 children had varying degrees of developmental delay, with 3 presenting solely developmental delay. EEG abnormalities were present in all the 21 children with seizures, including focal or multifocal discharges in 20 children, generalized discharges in 10 children, hypsarrhythmia in 2 children, and electrical status epilepticus during sleep in 3 children. Magnetic resonance imaging abnormalities were found in 5 of the 24 children. Among the 21 children with seizures, 57% (12/21) achieved seizure control.Conclusions:KCNB1 gene variants are predominantly de novo missense variation. Most affected children present with epilepsy, though some may exhibit only developmental delay. Epilepsy often begins before 2 years of age, with focal seizures being the most common type. About 80% of patients experience clustered seizures. Although most patients achieve seizure control, they still exhibit varying degrees of developmental delay, consistent with developmental epileptic encephalopathy.

Objective This study aimed to explore the differences in global and local brain network topological properties between deficient pattern(DP)and excess pattern(EP)of mild vascular cognitive impairment caused by subcortical small vessel disease based on graph theory network.Methods Patients were recruited prospectively and were classified with DP and EP subtype.The global small-world topological attributes and local nodes were calculated for the comparison of DP,EP,and healthy controls(CN)using the GRETNA platform.Results The three groups all had small-world attributes,but only the patients in EP had a significantly lower small world attribute δ in the range of 0.05-0.26 than the control group(P<0.05).The node efficiency and node strength indicators of multiple brain region were able to significantly distinguish the DP group from the EP group.However,there was no positive brain region in the node efficiency of the DP patients(P>0.05),and only a few brain regions showed increased node strength efficiency(P<0.05).Conclusion The results indicate that the syndrome of DP and EP have significantly different neuroimaging phenotypes,providing a basis for further research of biological classification based on Chinese Medicine syndromes.

Objective:To investigate the effect of panax notoginseng saponins(PNS)on the expression of tumor necrosis factor-α(TNF-α)in lipopolysaccharide(LPS)-induced activated BV2 microglia through p38 mitogen-activa-ted protein kinase(p38 MAPK)pathway.Methods:BV2 microglia were divided into control group,LPS activated group and LPS+panax notoginseng saponins intervention group(LPS+PNS).The CCK-8 method was used to detect the viability of BV2 microglia and determine the optimal drug intervention concentration.Western Blot and immunofluo-rescence were used to detect the expression of p38 MAPK and TNF-α and the phosphorylation level of p38 MAPK(p-p38 MAPK)in BV2 microglia.Results:Compared with the blank control group,there was no significant difference in the cell viability of BV2 microglia,and finally 100 mg/L was selected as the drug intervention concentration.Western Blot and immunofluorescence results indicated that after LPS activation,the expression of TNF-α and the phosphoryla-tion level of p38 MAPK in BV2 microglia were significantly increased(P＜0.05).After PNS intervention,compared with LPS-activated group,the expression of TNF-α and the phosphorylation level of p38 MAPK were significantly decreased(P＜0.05).After treatment with p38 MAPK pathway inhibitor(SB203580),there was no significant differ-ence in the expression levels of p-p38 MAPK and TNF-α in PNS combined with SB203580 group(LPS+PNS+I)com-pared with LPS+PNS group(P＞0.05).In addition,the changes of p38 MAPK in each group were not statistically sig-nificant(P＞0.05).Conclusion:PNS may inhibit the expression of inflammatory factor TNF-α secreted by activated BV2 microglia through p38 MAPK pathway.

Objective:To investigate the clinical value of tissue Doppler echocardiography in the evaluation of left ventricular function in patients with pregnancy-induced hypertension.Methods:This is a case-control study, including 100 patients with pregnancy-induced hypertension who received treatment at the Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from May 2019 to May 2022, and an additional 100 healthy pregnant women who underwent physical examination during the same period. All participants underwent two-dimensional echocardiography, pulsed Doppler echocardiography, and tissue Doppler echocardiography. Ultrasound parameters related to left ventricular morphology and function were collected from all participants. The ultrasound parameters related to left ventricular morphology and function between patients with different types of pregnancy-induced hypertension and healthy controls were compared. The correlation between left ventricular function ultrasound parameters and serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels was investigated.Results:Patients with pregnancy-induced hypertension, patients with mild preeclampsia, patients with severe preeclampsia, and healthy controls demonstrated differences in interventricular septum thickness during diastole [(10.24 ± 1.18) mm, (11.39 ± 1.24) mm, (11.57 ± 1.29) mm, (8.81 ± 0.95) mm], left ventricular end-diastolic diameter [(47.31 ± 2.81) mm, (49.82 ± 2.89) mm, (52.03 ± 2.94) mm, (46.82 ± 2.76) mm], left ventricular posterior wall thickness [(9.73 ± 1.06) mm, (10.62 ± 1.13) mm, (11.75 ± 1.21) mm, (8.96 ± 0.97) mm], left ventricular inner diameter [(32.82 ± 2.34) mm, (35.48 ± 2.39) mm, (36.04 ± 2.45) mm, (30.41 ± 2.27) mm], and left ventricular mass index [(98.41 ± 7.83) g/m 2, (105.73 ± 8.26) g/m 2, (108.63 ± 8.57) g/m 2, (96.59 ± 7.69) g/m 2]. All of these parameters showed significant differences between patients with different types of pregnancy-induced hypertension and healthy controls ( F = 13.47, 12.61, 16.59, 13.26, 19.73, all P < 0.001). Significant differences were also observed in echocardiographic indices of left ventricular function such as peak velocity ratio of E and A waves, systolic motor amplitude, early peak diastolic velocity to late peak diastolic velocity, and Tei index between patients with different types of pregnancy-induced hypertension and healthy controls ( F = 12.84, 11.27, 14.64, 21.43, all P < 0.001). In patients with pregnancy-induced hypertension, peak velocity ratio of E and A waves, systolic motor amplitude, and early peak diastolic velocity to late peak diastolic velocity were moderately negatively correlated with serum NT-proBNP level ( r = -0.56, -0.43, -0.54, P = 0.029, 0.042, 0.031), while Tei index showed a positive correlation with serum NT-proBNP level ( r = 0.77; P = 0.003). Conclusion:Two-dimensional echocardiography, pulsed Doppler echocardiography combined with tissue Doppler echocardiography can be used to effectively evaluate the changes in left ventricular structure and function in patients with different types of pregnancy-induced hypertension. Monitoring the Tei index using tissue Doppler echocardiography can accurately reflect myocardial injury and functional changes, which has a great clinical application value.