Analysis and study of the mechanism of narcotic addiction and withdrawal.

Narcotic drugs refer to drugs that have anesthetic effects on the central nervous system, and they easily produce physical dependence and mental dependence and can be addictive due to continuous use, abuse or unreasonable use. In this paper, bioinformatics and data analysis and mining techniques were used to analyze the methylation differences in transcriptional and clinical data of narcotic addiction in public databases, to explore the mechanism of narcotic addiction, and to mine some norepinephrine drugs. This study confirmed the possibility of using norepinephrine as an auxiliary drug for drug addiction rehabilitation. In addition, we also conducted a similar analysis on the addiction of three drugs. The results showed that the differences in the body caused by the ingestion of opiates and cocaine were significantly greater than those caused by the ingestion of methamphetamine.

Prediction of immune and targeted drug efficacy in pain-related risk subtypes for bladder cancer patients.

Bladder cancer is a complex disease with high morbidity and mortality rates. At least 430,000 cases are diagnosed annually worldwide. Cancer pain is the most common and distressing symptom in cancer patients. Studies have reported depression, anxiety, and decreased quality of life in survivors of various cancers. The study of pain-related genes in cancer patients may provide a basis for developing targeted drugs for cancer therapy, which could reduce pain and improve quality of life of cancer patients. In this study, the mRNA expression and clinical data of bladder cancer patients were downloaded from public databases. A total of 103 pain-related genes were also downloaded from the public databases. Univariate Cox regression analysis identified 17 pain-related genes that were significantly associated with overall survival. We calculated a pain-related risk score for each patient, constructed a bladder cancer pain risk model, and categorized bladder cancer patients into two risk subtypes. Differences in prognosis, differential gene expression, immune cell signatures, hallmarks, metabolic pathways, and somatic mutations between the different risk subtypes were systematically investigated. Eight drugs associated with bladder cancer risk subtypes were identified. Their differences in the high- and low-risk subtypes of bladder cancer were examined. In addition, the response to immunotherapy was analyzed in patients with different pain-related subtypes. Results revealed significant differences in these characteristics. Finally, a predictive model for pain-related risk subtypes in patients with bladder cancer was established. The study findings provide a reference for prognostication and personalized medical treatment of bladder cancer patients.

Sufentanil inhibits the metastasis and immune response of breast cancer via mediating the NF-κB pathway.

OBJECTIVE: Breast cancer (BC) causes cancer-related death in women. Sufentanil is used for cancer pain and postoperative analgesia. This study aimed to explore the role of sufentanil in BC. METHODS: BC cells were treated with sufentanil, and cell viability was evaluated using the cell counting kit-8 (CCK-8) assay. Biological behaviors were analyzed using EDU assay, flow cytometry, transwell assay, western blotting, and ELISA. The levels of NF-κB pathway-related factors were examined using western blotting. A xenograft tumor model was established to assess the effects of sufentanil on tumor growth in vivo. RESULTS: Sufentanil at the concentration of 20, 40, 80, and 160 nM suppressed cell viability (IC50 = 39.84 in MDA-MB-231 cells, and IC50 = 47.46 in BT549 cells). Sufentanil inhibited the proliferation, invasion, epithelial-mesenchymal transition (EMT), and inflammation, but induced apoptosis of BC cells. Mechanically, sufentanil suppressed the activation of the NF-κB pathway. Rescue experiments showed that RANKL (NF-κB receptor agonist) abrogated the effects induced by sufentanil. Moreover, sufentanil inhibited tumor growth, inflammatory response, but promoted apoptosis via the NF-κB pathway in vivo. CONCLUSIONS: Sufentanil decelerated the progression of BC by regulating the NF-κB pathway, suggesting sufentanil may be used in BC therapy.

Sufentanil treatment inhibited BC cell proliferation, invasion, epithelial-mesenchymal transition (EMT), immune response, but induced apoptosis.Sufentanil suppressed the activation of the NF-κB pathway.RANKL (NF-κB receptor agonist) abrogated the effects induced by sufentanil.Sufentanil inhibited tumor growth, proliferation, immune response and promoted apoptosis via the NF-κB pathway.

Cannabinoid Receptor Type 2 Agonist Reduces Morphine Tolerance via Mitogen Activated Protein Kinase Phosphatase Induction and Mitogen Activated Protein Kinase Dephosphorylation.

Morphine is an opioid drug often used in treating moderate to severe pain. However, morphine tolerance in patients limits its used in clinical settings. Our previous study showed that a cannabinoid type 2 (CB2) receptor agonist attenuated morphine tolerance. However, the exact mechanism by which CB2 agonists reduce morphine tolerance remains unclear. In this study, we investigated the effect of mitogen activated protein kinase (MAPK) and mitogen activated protein kinase phosphatases 1 and 3 (MKP-1 and MKP-3) on the regulation of morphine tolerance by CB2 receptor agonist. Chronic morphine treatments for 7 days reduced the protein expression of MKP-1 and MKP-3 in the spinal cord and increased the phosphorylation of p38, ERK1/2 and the level of proinflammatory mediator, such as IL-1ß, IL-6 and TNF-α. Coadministration of CB2 receptor agonist AM1241 alleviated the inhibition of MKP-1 and MKP-3 by chronic morphine administration and reduced the expression of phosphorylated MAPK and proinflammatory factors. The effect of the CB2 receptor agonist on morphine-induced downregulation of MKP-1 and MKP-3 was reversed by the MKP-1 and MKP-3 antagonist triptolide. Our findings suggested that CB2 receptor agonist may induce the expression of MKP-1 and MKP-3 to promote MAPK dephosphorylation and reduce the production of downstream cytokine, thereby reducing morphine tolerance. This finding suggested that MKPs may serve as a new target for alleviating morphine tolerance.

Long Noncoding RNA WDFY3-AS2 Represses the Progression of Esophageal Cancer through miR-18a/PTEN Axis.

BACKGROUND: Understanding the role of lncRNAs in the development of human malignancies is necessary for the targeted therapy of malignant tumors, including esophageal cancer (EC). Nevertheless, the specific role and regulatory mechanism of lncRNA WDFY3-AS2 in EC are still unclear. Here, we examined the functional role and regulatory mechanism of WDFY3-AS2 in EC. MATERIALS AND METHODS: RT-qPCR assay was applied to measure the expression of WDFY3-AS2 and miR-18a in EC samples and cells. The luciferase reporter and RIP assays were used to check the relationship between WDFY3-AS2, miR-18a, and PTEN. Counting Clock Kit-8 (CCK-8) assay was carried out to detect cell viability, and transwell assays were used for measuring cell migration and invasion. RESULTS: Underexpression of WDFY3-AS2 was found in EC specimens and cells, which predicted a poor prognosis of EC patients. Reexpression of WDFY3-AS2 repressed the progression of EC via inhibiting cell proliferation, migration, and invasion. Additionally, WDFY3-AS2 was negatively correlated with miR-18a and positively with PTEN. Furthermore, we discovered that the expression of PTEN decreased by miR-18a mimic was rescued by WDFY3-AS2 overexpression. CONCLUSIONS: WDFY3-AS2 modulates the expressional level of PTEN as a competitive endogenous RNA via sponging miR-18a in EC, which suggests that the WDFY3-AS2/miR-18a/PTEN pathway might be involved in the progression of EC.

Caspase-1 inhibition ameliorates murine acute graft versus host disease by modulating the Th1/Th17/Treg balance.

Our previous studies have implicated Caspase-1 signaling in driving the proinflammatory state of acute graft versus host disease (aGVHD). Therefore, we aimed to elucidate the mechanism of Caspase-1 in in murine models of aGVHD through specific inhibition of its activity with the decoy peptide Ac-YVAD-CMK. We transplanted bone marrow from donor C57BL/6 (H-2b) mice into recipient BALB/c (H-2Kd) mice and randomized the recipients into the following treatment cohorts: (1) allogeneic hematopoietic stem cell transplantation and splenic cell infusion control (PBS group); (2) low dose Ac-YVAD-CMK (AC low group); (3) and high dose Ac-YVAD-CMK (AC high group). Indeed, we observed that Caspase-1 inhibition by Ac-YVAD-CMK ameliorated pathological damage and inflammation in the liver, lungs, and colon elicited by aGVHD. This was associated with reduced mortality secondary to aGVHD. Mechanistically, we found that Caspase-1 inhibition modulated donor T cell expansion, restored the balance of Th1/Th17/Treg subsets, and markedly decreased serum levels and aGVHD target organ mRNA expression of IL-1ß, IL-18, and HMGB1. Thus, we demonstrate that inhibition of Caspase-1 by Ac-YVAD-CMK mitigates murine aGVHD by regulating Th1/Th17/Treg balance and attenuating its characteristic proinflammatory state.

CircRNA circUGGT2 Contributes to Hepatocellular Carcinoma Development via Regulation of the miR-526b-5p/RAB1A Axis.

BACKGROUND: Hepatocellular carcinoma (HCC) is a common malignant tumor in the world. Circular RNA hsa_circ_0008274 (circUGGT2) is reported to be upregulated in HCC tissues. Notwithstanding, the role and regulatory mechanism of circUGGT2 in HCC are indistinct. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was implemented to examine the levels of circUGGT2, microRNA (miR)-526b-5p, and ras-related protein Rab-1A (RAB1A) mRNA in HCC tissues and cells. Cell proliferation and colony formation were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) or colony formation assays. The levels of cyclin D1, proliferating cell nuclear antigen (PCNA), and RAB1A were detected with Western blotting. Cell cycle progression, migration, and invasion were evaluated by using flow cytometry or transwell assays. The relationship between circUGGT2 or RAB1A and miR-526b-5p was verified via dual-luciferase reporter and/or RNA pull-down assays. Xenograft assay was executed to confirm the role of circUGGT2 in vivo. RESULTS: We observed that circUGGT2 and RAB1A were upregulated while miR-526b-5p was downregulated in HCC tissues and cells. CircUGGT2 silencing suppressed tumor growth in vivo and curbed proliferation, colony formation, cell cycle progression, migration, and invasion of HCC cells in vitro. Mechanically, circUGGT2 regulated RAB1A expression via competitively binding to miR-526b-5p. Also, the inhibitory influence of circUGGT2 silencing on the malignancy of HCC cells was overturned by miR-526b-5p inhibitor. Furthermore, RAB1A overexpression reversed the suppressive influence of miR-526b-5p mimic on the malignancy of HCC cells. CONCLUSION: CircUGGT2 silencing inhibited HCC development via modulating the miR-526b-5p/RAB1A axis, providing a possible target for HCC treatment.

[Caspase1 Inhibitor Ac-YVAD-CMK Prevents and Treats the Acute Graft Versus Host Disease in Mice].

OBJECTIVE: To explore the effect of Caspase 1 inhibitor Ac-YVAD-CMK on acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation(allo-HSCT) and its mechanism. METHODS: Experiments were divided randomly into 3 groups: allogeneic hematopoietic stem cell transplantation combined with splenic cell infusion group (TS group, n=12), allogeneic hematopoietic stem cell transplantation combined with splenic cell infusion plus injection of low dose Caspase 1 inhibitor group (TS+low dose of C group, n=16) and plus high dose Caspase1 inhibitor (TS+high dose of C group, n=19). The body weight of mice in each group was dynamically detected, and the clinical manifestation of GVHD and score of aGVHD were determined, and the chimerism rate of mice was detected after transplantation for 14 days. Th1, Th2 and Th17 cells in peripheral blood were examined by flow cytometry. Peripheral proinflammatory cytokines IL-1ß, IFN-γ, IL-1α and IL-18 were examined by enzyme-linked immunosorbent assay(ELISA). The tissues sections of GVHD target organs (liver, lung, colon and skin) were stained with HE for histopathologic examination and histopathologic score. RESULTS: Ac-YVAD-CMK could alleviate murine aGVHD and pathological injury, decreare the incidence and severity of aGVHD in recipient mice. The detection of Th cell subsets in peripheral blood by flow cytometry showed that compared with TS group, the Th1 cell ratio in TS+low dose of C and TS+high dose of C groups was significantly reduced (P<0.05), while the Th2 and Th17 cell ratio was significantly enhanced (P<0.05) in TS+low dose of high dose of C groups. The detection of peripheral inflamematory cytokines by ELISA demonstrated that the inflammatory cytokines including IL-1ß,IFN-γ,IL-18 and IL-1α were reduced significantly (P<0.05). CONCLUSION: Ac-YVAD-CMK can improve aGVHD by inhibiting Caspase 1 and reducing the release of some inflammatory cytokines, thereby alleviated the aGVHD pathological damage.

Synthesis and Luminescence Properties of Terbium-Doped Lanthanum Oxychloride Nanostructures.

LaOCl:Tb3+ nanofibers, nanotubes and nanobelts were prepared via electrospinning combined with a double-crucible chlorination technique using NH4Cl powders as chlorinating agent. Different morphologies of LaOCl:Tb3+ nanostructures were obtained through adjusting some of the electrospun parameters. The as-prepared LaOCl:Tb3+ nanostructures are tetragonal in structure with space group of P4/nmm. The diameters of LaOCl:Tb3+ nanofibers, nanotubes and the width of LaOCl:Tb3+ nanobelts are respectively 133.99 ± 16.95 nm, 140.57 ± 17.82 nm and 5.32 ± 0.63 µm under the 95% confidence level. Under the excitation of 230-nm ultraviolet light, the LaOCl:Tb3+ nanostructures emit the predominant emission peaks at 544 nm originated from the energy levels transition of 5D4 --> 7F5 of Tb3+ ions. The optimum molar percentage of Tb3+ in the LaOCl:Tb3+ nanofibers is 7%. LaOCl:Tb3+ nanobelts exhibit the strongest PL intensity of the three nanostructures under the same doping molar concentration. The possible formation mechanisms of LaOCl:Tb3+ nanostructures are also proposed.

Expression of PEBP4 protein correlates with the invasion and metastasis of colorectal cancer.

This study aimed to investigate the expression of PEBP4 protein in colorectal carcinoma tissues and its correlation with the clinical pathology of colorectal cancer and to investigate the relationship between PEBP4 expression and the invasion and metastasis of colorectal cancer cells, which could provide an experimental basis for future biological treatments of human colorectal cancer. RT-PCR and western blot methods were applied to detect the mRNA and protein expressions, respectively, of PEBP4 in colorectal cancer tissues and normal pericarcinoma tissues, and their correlations with the tumorigenesis and development of colorectal cancer, as well as its clinical pathology, were analyzed. Using the RNA interference technology, the expression of PEBP4 was knocked down in the human colorectal cancer cell HCT116, and the changes of the invasion capability of HCT116 were monitored. The positive mRNA expression rate of PEBP4 in colorectal cancer tissue was significantly higher than that in the normal pericarcinoma tissue (p < 0.05). Also, the positive expression rate in the cancer tissues from patients with positive lymph node and distant metastasis was significantly higher than that from the patients negative for lymph node and distant metastasis (p < 0.05). The positive expression rate of PEBP4 in the cancer tissues from the patients in early stages (I, II) was significantly lower than the expression rate in patients in advanced stages (III, IV) (p < 0.05). A lower degree of differentiation in colorectal cancer corresponded to a higher positive mRNA expression rate of PEBP4 (p < 0.05). However, this was independent of the patient's gender, age, and tumor size (p > 0.05). In colorectal cancer tissue, the expression of PEBP4 protein was consistent with its mRNA. Namely, PEBP4 protein expression in colorectal cancer tissues was significantly higher than that in the normal pericarcinoma tissues (p < 0.05), the expression in the cancer tissues from the patients with positive lymph node and distant metastasis was significantly higher than that from the patients who were negative for these metastases (p < 0.05), and a lower degree of differentiation in colorectal cancer corresponded to a higher TNM staging along with a higher PEBP4 protein expression (p < 0.05). After HCT116 cells transfected with PEBP4 siRNA, they showed a significantly lower expression level of PEBP4 protein (p < 0.05), and the number of cells that passed through the Transwell chamber was significantly lower compared to the non-transfected or the transfected controls (p < 0.05). The over-expression of PEBP4 protein may be related to the tumorigenesis, development, metastasis, and invasion of colorectal cancer.