Objective To explore the mechanism by which zinc-regulated transporters, iron-regulated transporter-likeprotein 4 (ZIP4) regulates glycolysis and its impact on tumor progression in cholangiocarcinoma (CCA), providing a theoretical basis for targeted therapy of CCA. Methods ZIP4 expression in CCA was analyzed using the GEPIA database. Immuno-histochemistry (IHC) was used to detect ZIP4 expression in 20 paired CCA and adjacent non-tumor tissues. Stable ZIP4-overexpressing CCA cell lines (ZIP4-OE) were established. Gene set enrichment analysis was used to screen differentially expressed genes and pathways in ZIP-OE CCA cells. ZIP4, N-myc proto-oncogene protein (MYCN), and histone-lysine N-methyltransferase 2E (KMT2E) were knocked down using small interfering RNAs (siRNAs). The expression of glycolysis-related gene (glucose transporter 1 [Glut1], hexokinase 2 [HK2], and lactate dehydrogenase A [LDHA]) was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glycolytic activity was assessed by measuring the extracellular acidification rate (ECAR). Cell proliferation was evaluated using colony formation assays, and cell migration was assessed using Transwell assays. A xenograft mouse model was constructed to examine CCA tumor growth. Protein levels of ZIP4, KMT2E, H3K4me3 (tri-methylation of lysine 4 on histone H3), and MYCN were detected by Western blotting. Results GEPIA database analysis and IHC results confirmed significantly higher ZIP4 expression levels in CCA tissues compared to adjacent non-tumor tissues (P＜0.05). Compared to the control group, the ZIP4-OE group exhibited a significantly increased ECAR, along with significantly enhanced proliferation and migration abilities (P＜0.01). Conversely, knockdown of ZIP4 suppressed CCA cells proliferation and migration. GEPIA analysis indicated that ZIP4 upregulates the transcription of oncogene MYCN, as well as glycolysis-related genes. Knockdown of MYCN abolished the ZIP4 overexpression-induced upregulation of Glut1, HK2, and LDHA gene transcription, reduced glycolysis, and significantly inhibited CCA cell proliferation and migration (P＜0.05). Mechanistic studies demonstrated that ZIP4 increases H3K4me3 level via KMT2E, leading to MYCN transcription. Knockdown of KMT2E in CCA cells suppressed the ZIP4 overexpression-induced enhancement in H3K4me3 modification, resulting in MYCN downregulation and significantly reduced CCA cells proliferation and migration (P＜0.05). Conclusions ZIP4 upregulates H3K4me3 modification through KMT2E, which recruits transcription factors to activate the transcription of MYCN. This subsequently enhances cellular glycolysis and promotes the proliferation and migration of CCA cells.

Objective:To investigate the phenotypic characteristics of dendritic cell (DC) in the peripheral blood of patients with chronic antibody-mediated rejection (CAMR) after kidney transplantation,revealing the potential role of DCs in the occurrence and development of CAMR.Methods:Case-control study. A total of 25 kidney transplant recipients who underwent kidney transplantation or were under long-term follow-up at West China Hospital of Sichuan University from October 1st, 2020, to July 31st, 2021, were recruited in this case-control study, comprising 15 cases of patients with CAMR and 10 cases of patients with stable renal function. Flow cytometry was used to analyze DC subsets in peripheral blood mononuclear cells (PBMC) of patients. Correlation analysis was conducted to explore the relationship between DC subsets and renal function indicators. Iterative cluster analysis was performed based on the absolute counts of DC, ILT3+ mDC, ILT3+ pDC,ILT4+ mDC and ILT4+ pDC in the CAMR group.Results:Compared with the stable renal function group, the percentages and absolute count of pDC, pDC/mDC ratio, and absolute count of ILT3+ pDC were decreased while percentages of ILT3+ mDC were increased in CAMR group . DC percentage was negatively correlated with Scr (r=-0.467, P=0.019) and CysC (r=-0.502, P=0.011) and positively correlated with eGFR (r=0.436, P=0.029). ILT4+ pDC percentage was positively correlated with CysC (r=0.433, P=0.031), and absolute count of DCs was negatively correlated with CysC (r=-0.425, P=0.034). Iterative cluster analysis defined two patient clusters with significant statistic differences of Scr, eGFR, and CysC detection results in the CAMR group.Conclusion:The distribution of DC subsets and the expression of ILT3 and ILT4 in the blood of post-transplant patients could provide new biomarkers for early identification of CAMR risk and optimization of immunosuppressive therapy strategies. Particularly, changes of pDC and the expression levels of ILT3, ILT4 can serve as valuable indicators for assessing post-transplant immune status and renal function prognosis.

Objective:To investigate the phenotypic characteristics of dendritic cell (DC) in the peripheral blood of patients with chronic antibody-mediated rejection (CAMR) after kidney transplantation,revealing the potential role of DCs in the occurrence and development of CAMR.Methods:Case-control study. A total of 25 kidney transplant recipients who underwent kidney transplantation or were under long-term follow-up at West China Hospital of Sichuan University from October 1st, 2020, to July 31st, 2021, were recruited in this case-control study, comprising 15 cases of patients with CAMR and 10 cases of patients with stable renal function. Flow cytometry was used to analyze DC subsets in peripheral blood mononuclear cells (PBMC) of patients. Correlation analysis was conducted to explore the relationship between DC subsets and renal function indicators. Iterative cluster analysis was performed based on the absolute counts of DC, ILT3+ mDC, ILT3+ pDC,ILT4+ mDC and ILT4+ pDC in the CAMR group.Results:Compared with the stable renal function group, the percentages and absolute count of pDC, pDC/mDC ratio, and absolute count of ILT3+ pDC were decreased while percentages of ILT3+ mDC were increased in CAMR group . DC percentage was negatively correlated with Scr (r=-0.467, P=0.019) and CysC (r=-0.502, P=0.011) and positively correlated with eGFR (r=0.436, P=0.029). ILT4+ pDC percentage was positively correlated with CysC (r=0.433, P=0.031), and absolute count of DCs was negatively correlated with CysC (r=-0.425, P=0.034). Iterative cluster analysis defined two patient clusters with significant statistic differences of Scr, eGFR, and CysC detection results in the CAMR group.Conclusion:The distribution of DC subsets and the expression of ILT3 and ILT4 in the blood of post-transplant patients could provide new biomarkers for early identification of CAMR risk and optimization of immunosuppressive therapy strategies. Particularly, changes of pDC and the expression levels of ILT3, ILT4 can serve as valuable indicators for assessing post-transplant immune status and renal function prognosis.

Objective:To investigate the relationship between the levels of serum cytokines and chemokines and the prognosis of patients with acute B-ALL after receiving chimeric antigen receptor (CAR)-T cell immunotherapy and acute graft-versus-host disease (aGVHD) in patients after bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:According to the case-control principle, Forty-two patients with B-ALL who received CD19-CAR-T cell immunotherapy bridged to allo-HSCT at Heibei Yanda Ludaopei Hospital from September 18, 2019 to May 9, 2022 were enrolled. Mann-Whitney U test was used to compare the changes of aGVHD-related cytokines and chemokine levels between CAR-T cell immunotherapy and bridging transplantation in different patients at the same time. Their plasma levels of cytokines and chemokines related to aGVHD were monitored at the day before CAR-T therapy and after CAR-T treatment at day 4, 7,14,21,28. The receiver operating characteristic curve was drawn to evaluate the predictive value of cytokines and chemokines in predicting the occurrence and the death of aGVHD patients. Kaplan-Meier method and Log-rank tests were used for Overall survival (OS) analysis. Results:Twenty-four of total 42 patients had aGVHD, of which 11 patients died and 31 patients survived. There was no significant difference in cytokines and chemokines between the aGVHD group and the non-aGVHD group on the day before CAR-T cell treatment. According to statistical analysis, the serum Elafin levels of aGVHD group was higher than that of non-aGVHD group at the 21st day [4 482 (2 811, 6 061) ng/L vs 2 466 (1 948, 3 375) ng/L, Z=3.145, P=0.001] and the 28st day [4 391 (2 808, 5594) ng/L vs 2 463 (1 658, 2 830) ng/L, Z=2.038, P=0.048] separately. At the 14th day, serum cytokines and chemokines levels between the two group were as follows,MIP-1 α [21.02 (12.36, 30.35) ng/L vs 5.56 (3.64, 10.79) ng/L], sCD25 [422.47 (257.99, 1 233.78) IU/ml vs 216.11 (133.75,457.39) IU/ml], Elafin [4 101 (2 393, 5 006) ng/L vs 2 155 (1 781, 3 033) ng/L], IL-6 [119.08 (23.97, 183.43) ng/L vs 8.39 (2.91, 17.42) ng/L] and IL-8 [13.56 (12.50, 24.52) ng/L vs 2.83 (1.73,6.87) ng/L] were at higher levels ( Z=2.653, P=0.007; Z=2.176, P=0. 030; Z=2.058, P=0.041; Z=3.329, P<0.001; Z=3.162, P=0.001). The KM survival curve showed that the cumulative survival rates of patients with higher serum levels of MIP-1α, sCD25, Elafin, IL-6 and IL-8 were lower than those with low levels at day 14, and the difference was statistically significant (χ 2=12.353, 4.890, 6.551, 10.563, 20.755, P<0.05). Conclusion:The outcomes of patients treated with CAR-T cell therapy bridged to allo-HSCT was correlated with serum MIP-1α, sCD25, Elafin, IL-6 and IL-8 levels after receiving CAR-T therapy. High concentrations of MIP-1α, sCD25, Elafin, IL-6 and IL-8 suggest poor prognosis and can be used as biomarkers to suggest appropriate clinical selection of therapy.

Induction of coronary collateral circulation, that is, therapeutic angiogenesis, is considered a promising treatment for coronary heart disease.However, coronary collateral growth is a complex process and is related to a variety of factors.Although it has achieved promising outcomes in animal experiments, clinical trials have so far failed to replicate these results.Further studies on the growth mechanisms of coronary collateral circulation are still needed before a feasible clinical treatment strategy becomes available.

Objective To establish the determination of luteolin and 1,7-dihydroxy-3,8-dimethoxyxanthone in Herbal Gentianopsis paludosa. Method The RP-HPLC with ZORBAX SB-C18 (250 mm?4.6 mm, 5 ?m) comlum was used. Mobile phase was methanol (A)-0.4% H3PO4 (B) Gradient elution was used, 0～15 min, A∶B=50∶50, 15～60 min, A∶B=55∶45. Detection wavelengh was 260 nm. Flow rate was 1.0 mL/min. Temperature of column was 30 ℃. Result The linear relationship of 1,7-dihydroxy-3,8-dimethoxyxanthone and luteolin were showed at the range of 0.084～0.84 ?g and 0.184～1.84 ?g (r=0.999 4 and r =0.999 7). The average recovery rate of 1,7-dihydroxy-3,8-dimethoxyxanthone and luteolin were 96.82% and 97.19%, RSD

0.05). In bone defects PeroGlas granales bond to bone and new bone formation were observed 4 weeks after operation. The bone defects were repa ired by new bone 12 weeks after PerioGlas implantation. Conclusion: PerioGlas may induce new bone formation in osseous defects.