OBJECTIVE To explore the effect of Jinkui shenqi pill and its potential mechanism on diabetic nephropathy-related osteoporosis （DNOP） in rats. METHODS The rats were randomly divided into blank control group， control group， model group， Jingui shenqi pill low-， medium- and high-dose groups （0.62， 1.24， 2.48 g/kg）， and positive control group （denosumab 3 mg/ kg）， with 12 cases in each group. Except for the blank control group and the control group， models of DNOP were constructed in the remaining groups by high-glucose and high-fat forage， 3% fructose solution for feeding+intraperitoneal injection of streptozotocin solution. At 2 d after successful molding， Jingui shenqi pill low-， medium- and high-dose groups， and positive control group were given intragastric administration or subcutaneous injection of the corresponding drugs， once a day， for consecutive 8 weeks. After the last administration， bone mineral density （BMD） and serum bone metabolism-related indexes [osteocalcin （OC）， calcium， phosphorus， alkaline phosphatase （ALP）， tartrate-resistant acid phosphatase （TRAP）] were detected. The pathological changes of right femoral tissues were observed， and the expressions of receptor activator of NF-κB ligand （RANKL）/receptor activator of NF-κB （RANK）/osteoprotegerin （OPG） pathway-related mRNAs and proteins in left femoral tissues were detected. RESULTS Compared with the blank control group and the control group， there was rupture of bone trabecula and severe structural damage in the model group， and tissue space was obvious. BMD and ALP activities， mRNA and protein expressions of OPG were significantly decreased or down-regulated， while the levels of OC， calcium and phosphorus， TRAP activity， mRNA and protein expressions of RANKL and RANK were significantly increased or up-regulated （P＜0.05）. Compared with the model group， the number of bone trabeculae was increased in all Jingui shenqi pill groups， and the arrangement of bone trabeculae was more regular and dense. The above indexes generally were improved in a certain dose-dependent manner （P＜0.05）. CONCLUSIONS Jingui shenqi pill can promote osteogenesis and regulate the balance of bone metabolism in DNOP rats， which may be related to down-regulating the expressions of RANKL and RANK， and up-regulating the expression of OPG.

BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

Objective:To investigate the clinical efficacy of daratumumab-containing regimen in the treatment of multiple myeloma (MM) and the associated factors affecting patients' progression-free survival (PFS).Methods:A retrospective case series study was conducted. Clinical data of 21 MM patients who were treated with daratumumab-containing regimen in the Heping Hospital Affiliated to Changzhi Medical College from January 2021 to September 2023 were collected. The patients were treated with daratumumab (16 mg/kg intravenous drip) combined with other drugs for 28 d as 1 cycle until disease progression. Among the 21 cases, 6 cases were newly diagnosed multiple myeloma (NDMM), 7 cases were relapsed/refractory multiple myeloma (RRMM), and 8 cases were second-line treatment with daratumumab after the poor outcome of VRD (bortezomib + lenalidomide +dexamethasone) regimen at the time of initial treatment (daratumumab second-line treatment group). The efficacy of the patients after 2 cycles of daratumumab treatment was summarized; the PFS of the whole group and the NDMM and RRMM patients was analyzed by using Kaplan-Meier method, and log-rank test was used for comparison between the groups; the different status of disease, gender and age were included in the univariate and multivariate Cox proportional hazards models to screen the factors affecting the PFS of MM patients.Results:The median age [ M ( Q1, Q3)] of 21 patients was 62 years old (55 years old, 68 years old); 17 were male and 4 were female. After 2 cycles of daratumumab treatment, the overall remission rate (ORR) of the whole group was 85.7% (18/21), 2 cases (9.5%) achieved strict complete remission (sCR), 3 cases (14.3%) achieved complete remission (CR), 9 cases (42.9%) achieved very good partial remission (VGPR), 4 cases (19.0%) achieved partial remission (PR), 2 cases (9.5%) had stable disease and 1 case (4.8%) had disease progression. After 2 cycles of daratumumab treatment, all 6 NDMM patients were in remission, with 2 cases of sCR, 1 case of CR, and 3 cases of VGPR; 4 of 7 RRMM patients were in remission, with 1 case of CR and 3 cases of PR; 8 patients with daratumumab second-line treatment were in remission, with 1 case of CR, 6 cases of VGPR, and 1 case of PR; the difference in ORR among the 3 groups was statistically significant ( P = 0.010), the difference in ORR between patients with NDMM and daratumumab second-line treatment was not statistically significant ( P = 0.245), the ORR of NDMM patients was higher than that of RRMM patients, and the difference was statistically significant ( P = 0.029). The median follow-up time was 15.4 months (95% CI: 13.7-17.1 months). The median PFS time for the whole group was 10.6 months (95% CI: 7.3-15.5 months); the median PFS time was not reached in NDMM patients, the median PFS time was 14.6 months (95% CI: 2.1-27.2 months) in RRMM patients, the median PFS time was 9.6 months (95% CI: 9.5-9.7 months) in patients with daratumumab second-line treatment, and the difference in PFS among the 3 groups was not statistically significant ( P = 0.085). Multivariate Cox regression analysis showed that high age was an independent risk factor for PFS in MM patients ( HR = 1.12, 95% CI: 1.03-1.21, P = 0.009). Conclusions:Daratumumab has good results in treating MM and can be used as a first-line treatment option for NDMM patients, which may improve the remission rate of MM patients with previous ineffective treatment of VRD regimen, and may also improve the prognosis of RRMM patients. High age may be a risk factor for disease progression in MM patients treated with daratumumab.

Objective:To systematically assess the effectiveness and safety of acupuncture for spastic hemiplegia after ischemic stroke. Methods:Randomized controlled trials(RCTs)of acupuncture treatment for spastic hemiplegia after ischemic stroke meeting the inclusion criteria in Cochrane Library,Medline,Excerpta Medica Database(EMBASE),PubMed,China National Knowledge Infrastructure(CNKI),SinoMed,Chongqing VIP Database(VIP),and Wanfang Data Knowledge Service Platform(Wanfang)published from each database's inception to February 2023 were retrieved by computer.Two reviewers independently extracted data and evaluated the risk of bias using Cochrane's risk of bias tool.Review Manager 5.4 was used for data analysis.Continuous data were evaluated using mean difference(MD)with a 95%confidence interval(CI),and dichotomous data were analyzed using risk ratio(RR). Results:A total of 24 trials,including 1 970 participants,were included in the study.The meta-analysis of 7 trials showed that compared to the rehabilitation therapy,acupuncture therapy was more effective in improving the simplified Fugl-Meyer assessment score after 1-month treatments[MD=10.52,95%CI(7.81,13.23),P<0.001].The meta-analysis of 2 articles showed the same tendency after 6-month treatments[MD=19.18,95%CI(11.34,27.02),P<0.001],and the 6-month treatment course resulted in better outcomes than the 1-month course.The meta-analysis of 8 trials showed that acupuncture had a better improvement on the Barthel index score than rehabilitation therapy after 1-month treatments[MD=10.78,95%CI(8.91,12.64),P<0.001].The meta-analysis of 2 articles showed the same tendency after 6-month treatments[MD=19.94,95%CI(19.02,20.87),P<0.001],and the 6-month course was better than the 1-month course.The meta-analysis of 2 trials showed that the effective rate of the modified Ashworth scale score improvement was more notable in the acupuncture group after 1-month treatments[RR=1.20,95%CI(1.02,1.40),P=0.020].One trial reported no adverse event,and 1 trial reported 3 adverse events without severe influence. Conclusion:Acupuncture might be an effective and safe therapy for spastic hemiplegia after ischemic stroke,but more high-quality,large-sample objectively-evaluated RCTs are needed to validate the conclusion.

The mesencephalic astrocyte-derived neurotrophic factor (MANF) has been recently identified as a neurotrophic factor, but its role in hepatic fibrosis is unknown. Here, we found that MANF was upregulated in the fibrotic liver tissues of the patients with chronic liver diseases and of mice treated with CCl₄. MANF deficiency in either hepatocytes or hepatic mono-macrophages, particularly in hepatic mono-macrophages, clearly exacerbated hepatic fibrosis. Myeloid-specific MANF knockout increased the population of hepatic Ly6C^high macrophages and promoted HSCs activation. Furthermore, MANF-sufficient macrophages (from WT mice) transfusion ameliorated CCl₄-induced hepatic fibrosis in myeloid cells-specific MANF knockout (MKO) mice. Mechanistically, MANF interacted with S100A8 to competitively block S100A8/A9 heterodimer formation and inhibited S100A8/A9-mediated TLR4-NF-κB signal activation. Pharmacologically, systemic administration of recombinant human MANF significantly alleviated CCl₄-induced hepatic fibrosis in both WT and hepatocytes-specific MANF knockout (HKO) mice. This study reveals a mechanism by which MANF targets S100A8/A9-TLR4 as a "brake" on the upstream of NF-κB pathway, which exerts an impact on macrophage differentiation and shed light on hepatic fibrosis treatment.

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine and metabolic disease, and its pathogenesis is closely related to inflammation, insulin resistance, and metabolic disorders. Bilirubin is the final product of the destruction and degradation of senescent red blood cells in the body. In addition, bilirubin can be not only used to evaluate liver function damage and cytotoxicity, but also can anti-inflammatory, antioxidant, alleviate metabolic disorders, etc. Recently, studies have found a certain correlation between low levels of bilirubin and PCOS: the level of bilirubin in patients with PCOS is low, and the anti-inflammatory and antioxidant properties of bilirubin may play a protective role in the pathogenesis of PCOS.

Objective:To investigate whether etomidate affects inflammatory response and apoptosis of PC12 cells induced by hypoxia by regulating miR-142-3p.Methods:PC12 cells were pretreated with different doses(2,6,12 μmol/L)of etomidate to establish hypoxia model;PC12 cells that transfected with miR-142-3p mimics or inhibitors were pretreated with 0 or 12 μmol/L of etomidate to establish hypoxia model.Cell viability,apoptosis and protein(CyclinD1,Cleaved-caspase-3)expressions were detected by CCK-8 method,flow cytometry and Western blot,respectively.ELISA was used to detect levels of inflammatory factors TNF-α,IL-1β,IL-6.Expression of miR-142-3p was detected by RT-qPCR.Results:Etomidate increased hypoxia-induced PC12 cells activity and expres-sion of CyclinD1 protein and miR-142-3p,while decreased cell apoptosis rate,Cleaved-caspase-3 protein expression and levels of inflammatory factors TNF-α,IL-1β,IL-6(P<0.05).Up-regulation of miR-142-3p increased activity and expression of CyclinD1 pro-tein of hypoxia-induced PC12 cells,while decreased cell apoptosis rate,Cleaved-caspase-3 protein expression and levels of inflamma-tory factors TNF-α,IL-1β,IL-6(P<0.05).Down-regulation of miR-142-3p reversed effects of etomidate on hypoxia-induced PC12 cell activity,apoptosis and expressions of inflammatory factors(P<0.05).Conclusion:Etomidate can reduce inflammatory response and apoptosis of PC12 cells induced by hypoxia,and its mechanism may be related to the up-regulation of miR-142-3p expression in cells.

Colorectal cancer (CRC), a malignant tumor worldwide consists of microsatellite instability (MSI) and stable (MSS) phenotypes. Although SHP2 is a hopeful target for cancer therapy, its relationship with innate immunosuppression remains elusive. To address that, single-cell RNA sequencing was performed to explore the role of SHP2 in all cell types of tumor microenvironment (TME) from murine MC38 xenografts. Intratumoral cells were found to be functionally heterogeneous and responded significantly to SHP099, a SHP2 allosteric inhibitor. The malignant evolution of tumor cells was remarkably arrested by SHP099. Mechanistically, STING-TBK1-IRF3-mediated type I interferon signaling was highly activated by SHP099 in infiltrated myeloid cells. Notably, CRC patients with MSS phenotype exhibited greater macrophage infiltration and more potent SHP2 phosphorylation in CD68⁺ macrophages than MSI-high phenotypes, suggesting the potential role of macrophagic SHP2 in TME. Collectively, our data reveals a mechanism of innate immunosuppression mediated by SHP2, suggesting that SHP2 is a promising target for colon cancer immunotherapy.

Objective:To evaluate the left ventricular myocardial work parameters of the animal models with left bundle branch block (LBBB) and evaluate the effects of LBBB on left ventricular function and motion pattern by pressure-strain loops (PSL) of speckle tracking imaging (STI).Methods:In Twenty-four healthy male beagles, LBBB was induced by radio frequency ablation under anesthesia, and blood pressure was measured at the same time. Electrocardiograms and echocardiography images were acquired before (baseline), 30 minutes after (acute-LBBB) and 3 months after(chronic-LBBB) the creation of LBBB respectively. STI was applied to measure the left ventricular global longitudinal strain (GLS) and obtain the the PSL of each time point to evaluate the left ventricular global and segmental myocardial work parameters.Results:Compared to the baseline, the global work efficiencies(GWE) were obviously reduced ( P<0.05) and global wasted works(GWW) were significantly increased ( P<0.01) in the acute-LBBB and chronic-LBBB, significant differences were observed in GLS between acute-LBBB and baseline( P=0.04). In baseline, the work efficiency (WEsept) and the constructive work (CWsept) in the basal and middle segments of the septal wall were both obviously higher than the corresponding segments of left ventricular lateral wall( P<0.01), while the distribution of the wasted work(WWsept) was opposite( P<0.01). In acute-LBBB, the WEsept of all segments were significantly decreased ( P<0.05), the WWsept were obviously increased( P<0.05), the CWsept of basal segment was significantly reduced( P=0.01), while the wasted work in the basal segment of lateral wall(WWlat) was increased( P=0.04) compared with the baseline. Compared with the acute-LBBB, the WEsept of basal and middle segments were mildly recovery( P=0.03) in chronic-LBBB, but were still lower than the baseline ( P=0.001), the changes of the other myocardial work parameters of septal and lateral wall were similar to the acute-LBBB. Conclusions:Both acute-LBBB and chronic-LBBB can lead to the changes of left ventricular global and segmental myocardial work parameters. The myocardial work parameters of left ventricle can quantitatively analyze the changes of left ventricular function and motion pattern of the LBBB.

Objective To investigate the protective effect of Carbachol on hyperoxia-induced acute lung injury (HALI) in mice and its related mechanisms. Methods Thirty-two healthy male ICR mice were randomly divided into four groups:control group, hyperoxia exposure three days group (HO3d group), hyperoxia exposure three days + Carbachol group (HO3d+Carba group), and Carbachol group (Carba group), eight mice in each group. The pathological changes of lung tissue in each group were observed under light microscope after the models were completed in each group.The expression of TLR4 and NF-κB protein in lung tissues were detected by Western blot, and the expression of HMGB-1 and TNF-α mRNA in lung tissues by RT-PCR. LSD-t test was used for sample pairwise comparison, and one-way ANOVA for intergroup comparison. P<0.05 was considered statistically significant. Results There was no statistical difference between the control group and the Carba group (P> 0.05), and no obvious abnormal changes in lung tissue structure. The expression of TLR4, NF-κB protein and HMGB-1 and TNF-α mRNA in the HO3d group were significantly higher than those in the control group (P<0.01), and there were obvious bleeding on the surface of the lung tissue and severe pathological damage. The expression of TLR4,NF-κB protein and HMGB-1 and TNF-α mRNA in the HO3d+Carba group were significantly lower than those in the HO3d group(P<0.01), while lung tissue damage degree was also lower than that in the HO3d group. Conclusions Hyperoxia can increase the expression of TLR4 and NF-κB in lung tissues, and cause inflammatory injury in lung tissue. Carbachol can reduce the release of HMGB-1 and TNF-α inflammatory factors in hyperoxia-induced acute lung injury, and its mechanism is related to the inhibition of TLR4/NF-κB signal pathway, which has a protective effect on HALI.