Myeloid-derived suppressor cells-induced exhaustion of CD8 + T-cell participates in rejection after liver transplantation.

Liver transplantation (LT) rejection remains the most pervasive problem associated with this procedure, while the mechanism involved is still complicated and undefined. One promising solution may involve the use of myeloid-derived suppressor cells (MDSC). However, the immunological mechanisms underlying the effects of MDSC after LT remain unclear. This study is meant to clarify the role MDSCs play after liver transplantation. In this study, we collected liver tissue and peripheral blood mononuclear cells (PBMC) from LT patients showing varying degrees of rejection, as well as liver and spleen tissue samples from mice LT models. These samples were then analyzed using flow cytometry, immunohistochemistry and multiple immunofluorescence. M-MDSCs and CD8 + T-cells extracted from C57/BL6 mice were enriched and cocultured for in vitro experiments. Results, as obtained in both LT patients and LT mice model, revealed that the proportion and frequency of M-MDSC and PD-1 + T-cells increased significantly under conditions associated with a high degree of LT rejection. Within the LT rejection group, our immunofluorescence results showed that a close spatial contiguity was present between PD-1 + T-cells and M-MDSCs in these liver tissue samples and the proportion of CD84/PD-L1 double-positive M-MDSC was greater than that of G-MDSC. There was a positive correlation between the activity of CD84 and immunosuppressive function of M-MDSCs including PD-L1 expression and reactive oxygen species (ROS) production, as demonstrated in our in vitro model. M-MDSCs treated with CD84 protein were able to induce co-cultured CD8 + T-cells to express high levels of exhaustion markers. We found that CD84 regulated M-MDSC function via expression of PD-L1 through activation of the Akt/Stat3 pathway. These results suggest that the capacity for CD84 to regulate M-MDSC induction of CD8 + T-cell exhaustion may play a key role in LT rejection. Such findings provide important, new insights into the mechanisms of tolerance induction in LT.

FLT3+ DC inhibits immune rejection via interaction with Treg in liver transplantation.

Fms-like tyrosine kinase 3 (FLT3) is a receptor tyrosine kinase (RTK) primarily expressed in hematopoietic stem cells and dendritic cells (DCs). While FLT3 plays a critical role in the proliferation, development and maintenance of DCs, thus influencing immune responses under both normal and pathological conditions, there also exists some evidence that FLT3+DC may be involved with immune responses in liver transplantation (LT). In this study, results from single-cell sequencing data analysis revealed a clear relationship between FLT3+DCs and Regulatory T cells (Tregs) in liver tissue of LT recipients. In peripheral blood samples of LT patients, levels of FLT3+DCs were decreased post-LT-surgery, while Tregs were increased. In a LT mouse model, levels of FLT3+DCs in the liver and bone marrow exhibited an initial time-dependent decrease followed by an increase after LT surgery. Results as obtained with co-culture experiments using mature BMDCs and CD4+ T cells revealed fluctuations in Tregs in response to FLT3 inhibitors and the FLT3 ligand. These findings suggest that FLT3+DCs could emerge as a novel target for mitigating immune rejection in LT.

Landscape and the immune patterns of cuproptosis in oral squamous cell carcinoma.

BACKGROUND: Oral squamous cell carcinoma is an increasingly prevalent cancer type characterized by high incidence and mortality rates. Its early detection is challenging, primarily because of the absence of early molecular markers. Cuproptosis is a novel regulatory mechanism of cell death with implications in various cancers. In this study, we aimed to study cuproptosis-related genes in oral squamous cell carcinoma to identify their prognostic value. METHODS: By analyzing genomic, bulk RNA-seq, and single-cell RNA-seq data, we investigated 13 cuproptosis-related genes in The Cancer Genome Atlas-Oral Squamous Cell Carcinoma dataset and Gene Expression Omnibus repository (GSE172577). RESULTS: ATP7A, ATP7B, and DLST were the most frequently mutated genes, with nine of our studied genes associated with overall survival. Single-cell analysis was conducted to identify cuproptosis-related tumor cells in oral squamous cell carcinoma, which revealed two distinct patterns based on the expression of cuproptosis-related genes. These patterns exhibit differences in genetic alterations and tumor immune microenvironment. Finally, we developed a cuproptosis index using a random forest algorithm based on cuproptosis pattern-related genes in which higher levels were linked to poorer prognosis. CONCLUSION: Our findings provide valuable insights into the mechanisms underlying oral squamous cell carcinoma-associated cuproptosis.

Characterization and Proteomic Analyses of Proinflammatory Cytokines in a Mouse Model of Liver Transplant Rejection.

Liver transplantation (LT) is an effective strategy for the treatment of end-stage liver disease, but immune rejection remains a significant detriment to the survival and prognosis of these LT patients. While immune rejection is closely related to cytokines, the cytokines investigated within previous studies have been limited and have not included a systematic analysis of proinflammatory cytokines. In the present study, we used a protein chip system and proteomics to detect and analyze serum proinflammatory cytokines and differentially expressed proteins in liver tissue in a mouse model of liver transplantation. In addition, bioinformatics analysis was employed to analyze the proinflammatory cytokines and differential changes in proteins in response to this procedure. With these analyses, we found that serum contents of GC-CSF, CXCL-1, MCP-5, and CXCL-2 were significantly increased after liver transplantation, while IL-5, IL-10, and IL-17 were significantly decreased. Results from Gene Ontology (GO) and KEGG pathway analyses revealed that the cytokine-cytokine receptor, Th1/Th2 cell differentiation, and JAK-STAT signaling pathway were enriched in a network associated with the activation of immune response. Results from our proteomic analysis of liver tissue samples revealed that 470 proteins are increased and 50 decreased, including Anxa1, Anxa2, Acsl4, Sirpa, S100a8, and S100a9. KEGG pathway analysis indicated that the neutrophil extracellular trap formation, NOD-like receptor signaling pathway, and leukocyte transendothelial migration were all associated with liver transplant rejection in these mice. Bioinformatics analysis results demonstrated that CXCL-1/CXCL-2 and S100a8/S100a9 were the genes most closely related to the functions of neutrophils and the mononuclear phagocyte system. These findings provide new insights into some of the critical factors associated with liver transplant rejection and thus offer new targets for the treatment and prevention of this condition.

MicroRNA-143 Increases Oxidative Stress and Myocardial Cell Apoptosis in a Mouse Model of Doxorubicin-Induced Cardiac Toxicity.

BACKGROUND Oxidative stress and myocardial apoptosis are features of doxorubicin-induced cardiac toxicity that can result in cardiac dysfunction. Previous studies showed that microRNA-143 (miR-143) was expressed in the myocardium and had a role in cardiac function. This study aimed to investigate the effects and possible molecular mechanisms of miR-143 on oxidative stress and myocardial cell apoptosis in a mouse model of doxorubicin-induced cardiac toxicity. MATERIAL AND METHODS Mice underwent intraperitoneal injection of doxorubicin (15 mg/kg) daily for eight days to develop the mouse model of doxorubicin-induced cardiac toxicity. Four days before doxorubicin administration, a group of mice was pretreated daily with a miR-143 antagonist (25 mg/kg/day) for four consecutive days by tail vein injection. The study included the use of a miR-143 antagomir, or anti-microRNA, an oligonucleotide that silenced endogenous microRNA (miR), and an agomir to miR-143, and also the AKT inhibitor, MK2206. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunoblot analysis were used to measure mRNA and protein expression, respectively. RESULTS Doxorubicin treatment increased the expression of miR-143, which was reduced by the miR-143 antagomir. Overexpression of miR-143 increased doxorubicin-induced myocardial apoptosis and oxidative stress. The use of the miR-143 antagomir significantly activated protein kinase B (PKB) and AKT, which were reduced in the presence of the AKT inhibitor, MK2206. However, the use of the miR-143 antagomir further down-regulated AKT phosphorylation following doxorubicin treatment and increased AKT activation. CONCLUSIONS In a mouse model of doxorubicin-induced cardiac toxicity, miR-143 increased oxidative stress and myocardial cell apoptosis following doxorubicin treatment by inhibiting AKT.

[Differences in root developmenly of winter wheat cultivars in Huang-Huai Plain, China].

Selecting one presently popularized winter wheat cultivar (Zhengmai 9023) and two cultivars (Abo and Fengchan 3) introduced in the 1950s and 1960s in Huang-Huai Plain as test materials, and by using minirhizotron technique, this paper studied the live root length, root diameter distribution, and net root growth rate of the cultivars. Fine roots with a diameter from 0.05 mm to 0.25 mm occupied the majority of the whole root system, and the fine roots with a diameter less than 0.5 mm accounted for 98% of the live root length. The average root diameter varied with plant growth, the variation range being 0.15 - 0.22 mm, and no significant difference was observe among the cultivars. The live root length was significantly positively correlated root number, suggesting that root number was the main factor for the increase of live root length. The most vigorous growth period of the roots was from reviving to jointing stage, and Abo and Fengchan 3 had a longer period increased root vitality, as compared with Zhengmai 9023. For Zhengmai 9023, its fine roots with a diameter more than 0.1 mm had an increasing proportion after jointing stage, which was helpful for improving plant resistance, root activity, and grain-filling at late growth stages.

[Effects of taurine on the ultrastructure and P2X7 receptor expression in brain following traumatic brain injury in rats].

OBJECTIVE: To explore the effects of taurine on the ultrastructure and P2X7 receptor protein expression in brain following traumatic brain injury (TBI) in rats. METHODS: Forty male SD rats, were divided randomly into four groups that were sham-operated group, TBI group, TBI plus low-dose taurine group and TBI plus high-dose taurine group. The TBI model was established by Marmarou's method, the expression of P2X7 receptor protein in parietal cortex and hippocampus was detected by the immunohistochemical method, the ultrastructure of parietal cortex were observed by transmission electron microscope. RESULTS: Compared with sham-operated group, the positive expression cells of P2X7 receptor protein in parietal cortex and hippocampus of TBI group were significantly increased (P < 0.01). Compared with TBI group, the positive expression cells of P2X7 receptor protein in parietal cortex and hippocampus of TBI plus low-dose taurine group and TBI plus high-dose taurine group were significantly decreased (P <0.01 or P <0.05). Compared with TBI plus low-dose taurine group, the positive expression cells of P2X7 receptor protein in parietal cortex and hippocampus of TBI plus high-dose taurine group were significantly decreased (P < 0.05 or P < 0.01). The pathological damage of parietal cortex in the TBI plus high-dose taurine group was obviously lightened. CONCLUSION: Taurine exerts the neuroprotective effect on TBI in rats, the protective mechanism might be associated with down-regulating the expression of P2X7 receptor protein in parietal cortex and hippocampus.

[Effect of progesterone on MMP-3 expression in neonatal rat brain after hypoxic-ischemia].

OBJECTIVE: To study the effect of progesterone on matrix metalloproteinase-3 (MMP-3) expression in neonatal rat brain after hypoxic-ischemia. METHODS: Followed the hypoxic-ischemia of neonatal rat brain, Evans blue (EB) staining and transmission electron microscopy were used to detect the blood-brain barrier pathological changes on permeability. MMP-3 protein expression in cerebral cortex was measured with Western blot. RESULTS: Transmission electron microscopy results showed that the blood brain barrier in hypoxic-ischemic group changed significantly compare to progesterone group. EB staining results suggested that the blood-brain barrier permeability of hypoxic-ischemic group was significantly increased compared to sham-operated group (P < 0.01). The blood-brain barrier permeability in progesterone group was also decreased in comparison to that of hypoxic-ischemic group (P < 0.05). Western blot image analysis results indicated that MMP-3 protein expression in the hypoxic-ischemic group increased significantly than that in sham-operated group (P < 0.01), and the progesterone group was decreased significantly than that in hypoxic-ischemic group (P < 0.05). CONCLUSION: Progesterone may reduce the blood-brain barrier damage by reducing MMP-3 expression. This might be one of the protective mechanisms in the hypoxic-ischemic brain injury.

Establishment of a human hepatoma multidrug resistant cell line in vitro.

AIM: To establish a multidrug-resistant hepatoma cell line (SK-Hep-1), and to investigate its biological characteristics. METHODS: A highly invasive SK-Hep-1 cell line of human hepatocellular carcinoma, also known as malignant hepatoma was incubated with a high concentration of cisplatin (CDDP) to establish a CDDP-resistant cell subline (SK-Hep-1/CDDP). The 50% inhibitory dose (IC(50)) values and the resistance indexes [(IC(50) SK-Hep-1/CDDP)/(IC(50) SK-Hep-1)] for other chemotherapeutic agents and the growth curve of cells were all evaluated using cell counting kit-8 assays. The distribution of the cell cycles were detected by flow cytometry. Expression of acquired multidrug resistance P-glycoprotein (MDR1, ABCB1) and multidrug resistance-associated protein 1 (MRP1, ABCC1) was compared with that in parent cells by Western blotting and immunofluorescence combined with laser scanning confocal microscopy. RESULTS: The SK-Hep-1/CDDP cells (IC(50) = 70.61 +/- 1.06 microg/mL) was 13.76 times more resistant to CDDP than the SK-Hep-1 cells (IC(50) = 5.13 +/- 0.09 microg/mL), and CDDP-resistant cells also demonstrated cross-resistance to many anti-tumor agents such as doxorubicin, 5-fluorouracil and vincristine. Similar morphologies were determined in both SK-Hep-1 and SK-Hep-1/CDDP groups. The cell cycle distribution of the SK-Hep-1/CDDP cell line exhibited a significantly increased percentage of cells in S (42.2% +/- 2.65% vs 27.91% +/- 2.16%, P < 0.01) and G2/M (20.67% +/- 5.69% vs 12.14% +/- 3.36%, P < 0.01) phases in comparison with SK-Hep-1 cells, while the percentage of cells in the G0/G1 phase decreased (37.5% +/- 5.05% vs 59.83% +/- 3.28%, P < 0.01). The levels of MDR1 and MRP1 were overexpressed in the SK-Hep-1/CDDP cells exhibiting the MDR phenotype. CONCLUSION: Multiple drug resistance of multiple drugs in the human hepatoma cell line SK-Hep-1/CDDP was closely related to the overexpression of MDR1 and MRP1.

Effect of tiaozhi yanggan decoction in treating patients with non-alcoholic fatty liver.

OBJECTIVE: To observe the clinical efficacy and safety of Tiaozhi Yanggan Decoction (TZYGD) in treating non-alcoholic fatty liver. METHODS: One hundred and thirty-eight patients were enrolled and randomized into two groups according to the random number table in a ratio of 3:1, with 8 cases eventually dropping out. The symptoms, signs, liver function markers, blood lipids, iconographic indices and clinical comprehensive efficacy after a 12-week treatment course were assessed in 101 patients treated with TZYGD in the treated group and 29 patients treated with Thiola in the control group. RESULTS: The total effective rate in the treated group and the control group was 81.19% and 68.97%, respectively, showing a significant difference between the two groups with the former being significantly higher than the latter (P<0.05). Moreover, the improvements in the symptoms, signs, liver function, blood lipids and iconographic indices in the treated group were favorable with no serious adverse reactions. CONCLUSION: TZYGD is effective and highly safe in treating non-alcoholic fatty liver.

[Observation of the effect of the mixed composite skin graft on deep partial thickness burn wounds].

OBJECTIVE: To evaluate the effect of mixed composite skin graft on the deep partial thickness burn wounds after tangential excision in burn patients. METHODS: Tangential excision was performed in 30 extremities of 23 burn patients within 3 postburn days (PBDs). Then large pieces of homologous acellular dermal matrix were grafted onto the superficial fascia with razor thin autoskin on top of them. The survival rate of skin grafts, the appearance and the functional recovery of the extremities were observed on 10 to 12 post operative day (POD). Skin samples from a healed wound of a patient were harvested three months after the injury for pathologic examination. RESULTS: The survival rate of the composite skin grafts was 93%. Necrosis was encountered in 7% of the grafts in the lower extremities due to the poor fixation of the grafts leading to separation of autologous skin and the dermal template, and also due to infection resulting in lysis of the grafts. The grafted skin was excellent in the appearance and elasticity, and function of the injured extremities recovered well after grafting after 3 - 6 months of follow-up. Epidermal and dermal texture was also good as shown by pathologic examination. CONCLUSION: Mixed composite skin grafting after early tangential excision might be an ideal and effective method in the management of deep partial thickness burn wounds.

[In vitro cleavage activity of the ribozymes constructed specifically for the second exons of pro alpha I and III collagen genes].

OBJECTIVE: To study the in vitro cleavage activity and the reaction conditions of the hammerhead ribozymes for the second exons of pro alpha 1I and III collagen genes, and observe the effect of the two ribozymes on collagen synthesis by the fibroblasts in scar tissue. METHODS: The fragments of the second exons of pro alpha 1I and III collagen genes were cloned into the plasmids pT-I and pT-III and labeled with (32)P during transcription to obtain the target RNA. The transcription products of the plasmids pT-gI and pT-gIII containing specific ribozymes were incubated with the label target RNAs, respectively, under various conditions, and the results observed by electrophoresis autoradiography. The constructed ribozymes were transduced into cultured fibroblasts via liposomes for investigation of their effects on mRNA synthesis of type I and III collagen protein by image analysis. RESULTS: The two ribozymes (rgI and rg III) could efficiently cleave their target RNAs at both 37 degrees Celsius and 42 degrees Celsius, in the presence of Mg(2+) within a relatively wide concentration range form 10 to 20 mmol/L. When the temperature was lowered from 65 degrees Celsius to 37 degrees Celsius and maintained at 37 degrees Celsius, the cleavage activity of the ribozymes reached the maximum. The expression of mRNA of I and III collagen decreased accompanied by reduced collagen synthesis. CONCLUSION: Hammerhead ribozymes for the fragments of the second exons of pro alpha 1I and III collagen genes can cleave its target RNAs in vitro and effectively inhibit the collagen synthesis by the scar-derived fibroblasts.