Anti-CD20 monoclonal antibodies for the treatment of refractory nephrotic syndrome （RNS） in children. The first- generation rituximab is the most widely used in clinical practice； it shows definite efficacy in children with RNS， is recommended by guidelines， particularly for achieving a high remission rate in minimal change nephrosis， and can significantly reduce the cumulative use of glucocorticoids and immunosuppressants. The second-generation ofatumumab has potential as an alternative treatment for patients who are intolerant or resistant to rituximab， while the third-generation obinutuzumab has shown efficacy in complex cases such as rituximab resistance or post-transplant recurrence. However， there is still controversy regarding the optimization of rituximab treatment dosage and whether ofatumumab and obinutuzumab offer greater advantages than rituximab for the treatment of RNS in children. The most common adverse reaction induced by anti-CD20 monoclonal antibodies is infusion reactions， and long-term adverse events mainly include increased risks of sustained immunosuppression and infections. Rituximab has significant economic advantages for the treatment of RNS， but additional pharmacoeconomic research based on China’s healthcare environment is needed to evaluate the cost-effectiveness of ofatumumab and obinutuzumab in this population. Given that the current use of ofatumumab and obinutuzumab in this field is considered off-label use， clinical application should only proceed after a rigorous evaluation of the patient’s benefits and risks.

Organoids are three-dimensional (3D) cellular structures formed through the differentiation and self-organization of pluripotent stem cells or tissue-derived cells, showing considerable potential in the research on disease mechanism, personalized medicine, and developmental biology. However, the development of organoids is limited by the complex composition, batch-to-batch variations, and immunogenicity of basement-membrane matrix in the current culture system, which hinders the clinical translation and in vivo applications of organoids. Hydrogels are highly hydrated 3D polymer network materials, with modifiable mechanical and biochemical properties by engineering, representing an ideal alternative to basement-membrane matrix. This article reviews the research progress in engineered hydrogels with defined composition currently used in organoid culture. We introduce the structural characteristics and engineering design considerations of hydrogels, emphasize the latest research progress and specific application cases, and discuss the future development of these engineered hydrogels, provide valuable insights for the further advancement and optimization of engineered hydrogels for organoid.
Hydrogels/chemistry* ; Organoids/cytology* ; Tissue Engineering/methods* ; Humans ; Animals ; Pluripotent Stem Cells/cytology* ; Cell Culture Techniques, Three Dimensional/methods* ; Tissue Scaffolds

Blood and lymphatic vessels are absent from the normal cornea.Pathological stimulation such as corneal in-fection,chemical burns and transplantation rejection can disrupt the balance between pro-and anti-lymphangiogenesis fac-tors,causing lymphatic vessels to extend from the corneal limbus to the central cornea.Corneal lymphangiogenesis is closely related to various regulatory factors and cellular signaling pathways.Further study and elucidation of the mechanism of corneal lymphangiogenesis will open up new directions for treating corneal transplantation rejection,inflammatory disea-ses,dry eyes and tumor metastasis.This review concludes the endogenous regulatory factors of corneal lymphangiogenesis and therapeutic strategies for its related ocular surface diseases.

Objective:To investigate the serum lactate level in patients with rheumatoid arthritis(RA)and its relationship with disease activity,and to analyze the effect of sodium lactate on the activation of CD4+T cells,the ability of secreting cytokines and CD4+T cell subsets in peripheral blood of the RA patients.Methods:The peripheral blood of healthy controls(HC)and RA patients was collected,and the content of lactate in the supernatant was detected by lactate detection kit,the correlation between the content of lactate and the disease score of the RA patients was analyzed;the activation level of CD4+T cells,the proportion of CD4+T cell subsets and the cytokines secreted by CD4+T cells in peripheral blood of all the RA patients were detected by flow cytometry after being stimulated with sodium lactate.Results:The serum lactate level in the RA patients(n=66)was significantly higher than that in the HC(n=60,P＜0.001),and there was a certain correlation with disease activity score in 28 joints(DAS28)-C-reactive protein(CRP)(r=0.273,P=0.029),The levels of rheumatoid factor[RF,197.50(26.03,783.00)IU/mL vs.29.30(0.00,102.60)IU/mL,P＜0.01],CRP[37.40(11.30,72.60)mg/L vs.5.83(2.36,12.45)mg/L,P＜0.001],were increased in patients with the lactate concentration greater than 5 mmol/L were significantly higher than those in patients with the lactate concentration less than or equal 5 mmol/L,however,there was no significant difference in the expres-sion of erythrocyte sedimentation rate[ESR,42.00(19.00,77.00)mm/h vs.25.00(12.50,45.50)mm/h,P＞0.05]and anti-cyclic citrullinated peptied(CCP)antibody[82.35(17.70,137.00)RU/mL vs.68.60(25.95,119.70)RU/mL,P＞0.05].Compared with the control group,the expression of PD-1(46.15％±8.54％vs.41.67％±9.98％,P＜0.001),inducible costimulatory molecule(ICOS,5.77％±8.60％vs.18.65％±7.94％,P＜0.01)and CD25(25.89％±5.80％vs.22.25％±4.59％,P＜0.01)on the surface of CD4+T cells in the RA patients treated with sodium lactate was significantly increased.Compared with the control group,the proportion of Th17(4.62％±1.74％vs.2.93％±1.92％,P＜0.05)and Tph(28.02％±6.28％vs.20.32％±5.82％,P＜0.01)cells in CD4+T cells of the RA patients in the sodium lactate treatment group increased.Compared with the con-trol group,the expression of IL-21(5.73％±1.59％vs.4.75％±1.71％,P＜0.05)inCD4+Tcells was up-regulated in the RA patients treated with sodium lactate.Conclusion:The level of serum lactate in RA patients is increased,which promotes the activation of CD4+T cells and the secretion of IL-21,and up-regulates the proportion of Th 17 and Tph cells in the RA patients.

Objective:To investigate frequency and function of peripheral CD4+CD28-T cells in immune thrombocytopenia(ITP)patients,to deeply understand pathogenesis of ITP.Methods:A total of 30 ITP patients treated in The Second Affiliated Hospi-tal of Dalian Medical University from January 2021 to June 2022 were recruited,and 30 gender/age-matched healthy volunteers were served as control group.Peripheral anticoagulation was collected and mononuclear cells(PBMC)were extracted,frequencies and func-tions of CD4+CD28-T cells in PBMC was analyzed by flow cytometry,plasma levels of IL-18 and IL-18BP in ITP patients were detected by ELISA,and frequencies of CD4+CD28-T cells before and after treated with IL-18 were determined by flow cytometry.Results:Frequencies of CD4+CD28-T cells in peripheral blood of ITP patients were significantly higher than that of healthy control group,and CD4+CD28-T cells had a stronger ability to secrete IFN-γ and TNF-α;level of IL-18 was higher in plasma from ITP patients;IL-18 could increase frequency of CD4+CD28-T cells in PBMC of ITP patients.Conclusion:Frequency of CD4+CD28-T cells in peripheral blood of ITP patients is significantly increased,and has stronger ability to secrete inflammatory factors.

BACKGROUND: Colorectal carcinogenesis and progression are related to the gut microbiota and the tumor immune microenvironment. Our previous clinical trial demonstrated that berberine (BBR) hydrochloride might reduce the recurrence and canceration of colorectal adenoma (CRA). The present study aimed to further explore the mechanism of BBR in preventing colorectal cancer (CRC). METHODS: We performed metagenomics sequencing on fecal specimens obtained from the BBR intervention trial, and the differential bacteria before and after medication were validated using quantitative polymerase chain reaction. We further performed ApcMin/+ animal intervention tests, RNA sequencing, flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assays. RESULTS: The abundance of fecal Veillonella parvula ( V . parvula ) decreased significantly after BBR administration ( P = 0.0016) and increased through the development from CRA to CRC. Patients with CRC with a higher V. parvula abundance had worse tumor staging and a higher lymph node metastasis rate. The intestinal immune pathway of Immunoglobulin A production was activated, and the expression of TNFSF13B (Tumor necrosis factor superfamily 13b, encoding B lymphocyte stimulator [BLyS]), the representative gene of this pathway, and the genes encoding its receptors (interleukin-10 and transforming growth factor beta) were significantly upregulated. Animal experiments revealed that V. parvula promoted colorectal carcinogenesis and increased BLyS levels, while BBR reversed this effect. CONCLUSION: BBR might inhibit V. parvula and further weaken the immunomodulatory effect of B cells induced by V. parvula , thereby blocking the development of colorectal tumors. TRIAL REGISTRAION ClinicalTrials.gov, No. NCT02226185.
Animals ; Humans ; Berberine/therapeutic use* ; Carcinogenesis ; Veillonella ; Colorectal Neoplasms/genetics* ; Tumor Microenvironment

Objective To explore the influence of family history of diabetes and central obesity on prediabetes(Pre-DM)and their interaction.Methods Based on the REACTION studyduring August December 2011 in Dalian,2587 individuals aged≥40 with normal blood glucose(BG)were divided into newly diagnosed Pre-DM(n=522)and BG normal group(NC,n=2065)during follow-up.The influence of family history of DM and central obesity on prediabetes and their interaction were analyzed.Results The age,SBP,DBP,BMI,WC,HDL-C,LDL-C,TG,ALT,GGT,FPG,2 hPG,HbA1c,FIns,HOMA-IR,SUA and the incidence of hypertension,fatty liver disease,dyslipidemia,overweight/obesity and central obesity in Pre-DM group were all higher than those in NC group(P＜0.05).Multivariate logistic regression analysis showed that family history of diabetes and central obesity were risk factors for Pre-DM.After adjustment,no significant multiplicative interaction was shown between diabetes family history and central obesity on the risk of Pre-DM(P=0.764).Quantitative analysis of interaction showed that RERI=0.110,S=1.112 and AP=0.053,suggesting that DM family history and central obesity have no significant additive interaction on the risk of Pre-DM.Conclusion The family history of DM and central obesity are both associated with the occurrence of Pre-DM,but there is no significant interaction.

Remote pharmaceutical care refers to the process that pharmacists provide pharmaceutical care to patients remotely through information technology. Remote pharmaceutical care in China starts late and develops slowly. Therefore ，this paper discusses the pharmaceutical care modes that pharmacists at home and abroad can provide under the remote mode by collecting literature. The results show that foreign remote pharmaceutical care starts early and is relatively mature. The service mainly included remote follow-up and intervention ，24-hour online prescription and order review ，24-hour online drug reorganization ，and guidance on rational drug use in remote areas or community hospitals. The service population covers patients with cardiovascular disease ， diabetes，asthma，AIDS and so on. Some hospitals have established an integrated pharmaceutical care system of “Internet+Medical Consortium”in China ，with which pharmacists can provide patients with pharmaceutical care such as remote follow-up and intervention，drug consultation and so on. With the promotion of telemedicine ，domestic pharmacists can gradually expand the scope of services ，expand pharmaceutical services such as remote consultation and remote popular science push ，and realize the sharing of high-quality pharmaceutical care for the whole people.

Objective:To evaluate the relationship between the euchromatic histone-lysine N-methyltransferase (G9a) and sodium-dependent activation of potassium channel (Slack) in the dorsal root ganglia (DRG) of rats with neuropathic pain (NP).Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 1 month, weighing 100-120 g, were divided into 4 groups ( n=12 each) by a random number table method: sham operation group (S group), vector plus sham operation group (VS group), vector plus NP group (VN group), and G9a CRISPR/Cas9 knockout plus NP group (GN group). Sham operation was performed at the age of 2 months in group S. In group VS, AAV5 1 μl was microinjected into L 4 and L 5 DRG at the age of 1 month, and sham operation was performed at the age of 2 months.In VN group and GN group, AAV5 and G9a CRISPR/Cas9 knockout plasmid 1 μl were microinjected into L 4 and L 5 DRG at the age of 1 month, and NP model was established by spinal nerve ligation (SNL) at the age of 2 months.Six rats in each group were selected to measure the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) before microinjection (T 0), before SNL (T 1), and at 3, 5 and 7 days after SNL (T 2-4). The animals were sacrificed after the last behavioral testing, the DRGs of lumbar segment (L 4, 5) were removed for determination of the expression of G9a, dimethylation of histone H3 at lysine 9(H3K9me2) and Slack (by Western blot). At 7 days after establishing the model, 6 rats from each group were selected to culture the primary DRG neurons.The frequency and amplitude of Slack current in DRG neurons and miniature excitatory post-synaptic currents (mEPSCs) in the spinal dorsal horn were measured by whole-cell patch-clamp technique. Results:Compared with group S, the TWL was significantly shortened and the MWT was decreased at T 2-4, the expression of G9a and H3K9me2 in the spinal dorsal horn was up-regulated, the expression of Slack was down-regulated, the amplitude and frequency of Slack currents in DRG neurons were decreased, and the frequency of mEPSCs was increased in group VN ( P<0.05), and no significant change was found in the parameters mentioned above in group VS ( P>0.05). Compared with group VN, the TWL was significantly prolonged and the MWT was increased at T 2-4, the expression of G9a and H3K9me2 in the spinal dorsal horn was down-regulated, the expression of Slack was up-regulated, the amplitude and frequency of Slack currents in DRG neurons were increased, and the frequency of mEPSCs was decreased in group GN ( P<0.05). Conclusion:The mechanism of NP is related to up-regulating the expression of G9a in DRG, thus inhibiting the expression and opening of Slack channels in rats.

Objective To analyze whether free fatty acids(FFA)are associated with anxiety and depression in elderly patients with chronic heart failure(CHF).Methods A cross-sectional study was conducted in patients by using questionnaires.According to the Zung Self-Rating Anxiety Scale (SAS) and the Self-Rating Depression Scale(SDS),126 patients were divided into the depression group (n=94),non-depression group (n =32),and anxiety group (n =48),and non-anxiety group (n =78).The difference in FFA levels between the two groups was analyzed.Moreover,the association of FFA with anxiety and depression was analyzed Results The incidence of anxiety in elderly patients with CHF was 38.1％(48 cases),and the incidence of depression was 74.6％ (94 cases).FFA levels were higher in both the anxiety and depression groups than in the non-anxiety and non-depression groups [(559.9 ± 256.2)μmol/L vs.(393.8 ± 147.8) μmol/L,(573.8 ± 264.7)μmol/L vs.(483.2 ± 225.2)μmol/L,P＜0.05 and 0.01)].The FFA level was positively correlated with SAS and SDS in elderly patients with CHF(r=0.214 and 0.275,P＜0.05 and 0.01).Multivariate regression analysis showed that the FFA level was a risk factor for elderly CHF patients with anxiety(β=0.074,95％CI:0.024～0.125,P=0.004)and depression(β=0.084,95％CI:0.022～0.145,P=0.008).Conclusions FFA levels are correlated with anxiety and depression in elderly patients with CHF.The FFA level is a risk factor for anxiety and depression in elderly patients with CHF.Psychological intervention should be carried out early in CHF patients with high levels of FFA to reduce the incidence of anxiety and depression.