Synthesis and biological evaluation of artemisinin derivatives as potential MS agents.

In this paper, a series of artemisinin derivatives were synthesized and evaluated. Studies have shown that IFN-γ produced by Th1 CD4+ T cells and IL-17A secreted by Th17 CD4+ T cells played critical roles in the treatment of multiple sclerosis. We used different concentrations of artemisinin derivatives to inhibit Th1 / Th17 differentiation in naive CD4+ T cells and to characterize IFN-γ / IL-17A in in vitro experiments. The preliminary screening results showed that ester compound 5 exhibited obvious inhibitory activities on Th1 and Th17 (IFN-γ decreased from 41% to 3% and IL-17A decreased from 24% to 8% at the concentration of 10 nM to 10 µM), and carbamate compounds also had obvious inhibitory activities against Th17 at high concentration. Moreover, we investigated the effect of compound 5 on myelin oligodendrocyte glycoprotein (MOG)-induced mice experimental autoimmune encephalomyelitis (EAE) model in vivo. 100 mg/kg compound 5 effectively reduced the disease severity of EAE compared with the vehicle group. This research revealed that compound 5 could be a promising avenue as potential MS inhibitor.

Prognostic Significance of Preoperative Integrated Liver Inflammatory Score in Patients with Hepatocellular Carcinoma.

BACKGROUND The Integrated Liver Inflammatory Score (ILIS), which includes 5 serum indicators (albumin, bilirubin, neutrophil count, alpha-fetoprotein [AFP], and alkaline phosphatase [ALP]), is a novel inflammation-based predictive model associated with poor survival in hepatocellular carcinoma (HCC) patients. Our study aimed to assess the prognostic value of ILIS in HCC patients undergoing radical hepatectomy and establish a nomogram and artificial neural network based on their ILIS scores. MATERIAL AND METHODS This multicenter retrospective study included patients from 2 institutions from 2007 to 2017. Independent risk factors associated with Recurrence-free survival (RFS) and overall survival (OS) were identified through univariate and multifactor analysis in the training and validation groups, respectively. Afterward, column line graphs and artificial neural networks (ANN) were constructed and validated using the validation group. RESULTS A total of 432 patients were included in this study (275 in the training group and 157 in the validation group). In both cohorts, ILIS was correlated with pathological features such as tumor size, degree of differentiation, Child-Pugh class classification, and BCLC staging. Moreover, ILIS was identified as an independent risk factor for OS. ILIS-based nomograms and artificial neural networks also showed the prognostic value of ILIS. CONCLUSIONS Preoperative ILIS is an independent and effective predictor of prognosis in HCC patients treated with radical hepatectomy, as shown by the fact that higher ILIS are associated with worse patient prognosis. We have also established nomograms and ANNs that predict HCC prognosis with high accuracy.

Phenotypic and functional characterization of tumor-derived endothelial cells isolated from primary human hepatocellular carcinoma.

AIMS: Tumor endothelial cells (TECs) have been investigated using human tumor xenografts in mice models. In order to provide pure human TECs for the updating of clinical anti-angiogenic cancer therapy, in the present study we established a protocol of purification of TECs derived from clinical hepatocellular carcinoma (HCC) and revealed the TEC features by in vitro and in vivo assays. METHODS: We isolated TECs from fresh surgical resections of HCC by magnetic-activated cell sorting and purified by flow cytometry sorting upon CD31 expression, referred to as ECDHCCs. Next, we identified cultured ECDHCCs by morphology, phenotype, genotype, and functional assays. RESULTS: The ECDHCCs appeared as Weibel-Palade bodies under electron microscopy. They expressed endothelial markers, such as CD31, CD105, and vascular endothelial growth factor receptor 2, and expressed the genes that are associated with pro-angiogenesis, especially vascular endothelial growth factor, epiregulin, and programmed cell death 10. Functionally, ECDHCCs were capable of tube formation, wound healing, and Transwell migration in vitro. These in vitro behaviors were validated by in vivo Matrigel plug assay in mice. Finally, comparison of ECDHCC with the Hep-G2 liver cancer cell line showed there was no similarity of phenotype or function between these two types of cells. CONCLUSIONS: Tumor endothelial cells derived from human HCC can be isolated and purified from clinical samples by flow cytometer. They have the endothelial phenotype and morphologic features and are capable of tube formation and migration. This study provides a useful model for researchers to study tumor angiogenesis and screening of candidate targets.

Gut microbes improve prognosis of Klebsiella pneumoniae pulmonary infection through the lung-gut axis.

Background: The gut microbiota plays a vital role in the development of sepsis and in protecting against pneumonia. Previous studies have demonstrated the existence of the gut-lung axis and the interaction between the gut and the lung, which is related to the prognosis of critically ill patients; however, most of these studies focused on chronic lung diseases and influenza virus infections. The purpose of this study was to investigate the effect of faecal microbiota transplantation (FMT) on Klebsiella pneumoniae-related pulmonary infection via the gut-lung axis and to compare the effects of FMT with those of traditional antibiotics to identify new therapeutic strategies. Methods: We divided the mice into six groups: the blank control (PBS), pneumonia-derived sepsis (KP), pneumonia-derived sepsis + antibiotic (KP + PIP), pneumonia-derived sepsis + faecal microbiota transplantation(KP + FMT), antibiotic treatment control (KP+PIP+PBS), and pneumonia-derived sepsis+ antibiotic + faecal microbiota transplantation (KP + PIP + FMT) groups to compare the survival of mice, lung injury, inflammation response, airway barrier function and the intestinal flora, metabolites and drug resistance genes in each group. Results: Alterations in specific intestinal flora can occur in the gut of patients with pneumonia-derived sepsis caused by Klebsiella pneumoniae. Compared with those in the faecal microbiota transplantation group, the antibiotic treatment group had lower levels of proinflammatory factors and higher levels of anti-inflammatory factors but less amelioration of lung pathology and improvement of airway epithelial barrier function. Additionally, the increase in opportunistic pathogens and drug resistance-related genes in the gut of mice was accompanied by decreased production of favourable fatty acids such as acetic acid, propionic acid, butyric acid, decanoic acid, and secondary bile acids such as chenodeoxycholic acid 3-sulfate, isodeoxycholic acid, taurodeoxycholic acid, and 3-dehydrocholic acid; the levels of these metabolites were restored by faecal microbiota transplantation. Faecal microbiota transplantation after antibiotic treatment can gradually ameliorate gut microbiota disorder caused by antibiotic treatment and reduce the number of drug resistance genes induced by antibiotics. Conclusion: In contrast to direct antibiotic treatment, faecal microbiota transplantation improves the prognosis of mice with pneumonia-derived sepsis caused by Klebsiella pneumoniae by improving the structure of the intestinal flora and increasing the level of beneficial metabolites, fatty acids and secondary bile acids, thereby reducing systemic inflammation, repairing the barrier function of alveolar epithelial cells, and alleviating pathological damage to the lungs. The combination of antibiotics with faecal microbiota transplantation significantly alleviates intestinal microbiota disorder, reduces the selection for drug resistance genes caused by antibiotics, and mitigates lung lesions; these effects are superior to those following antibiotic monotherapy.

Modification of Iron-Rich Phase in Al-7Si-3Fe Alloy by Mechanical Vibration during Solidification.

The plate-like iron-rich intermetallic phases in recycled aluminum alloys significantly deteriorate the mechanical properties. In this paper, the effects of mechanical vibration on the microstructure and properties of the Al-7Si-3Fe alloy were systematically investigated. Simultaneously, the modification mechanism of the iron-rich phase was also discussed. The results indicated that the mechanical vibration was effective in refining the α-Al phase and modifying the iron-rich phase during solidification. The forcing convection and a high heat transfer inside the melt to the mold interface caused by mechanical vibration inhibited the quasi-peritectic reaction: L + α-Al8Fe2Si → (Al) + ß-Al5FeSi and eutectic reaction: L → (Al) + ß-Al5FeSi + Si. Thus, the plate-like ß-Al5FeSi phases in traditional gravity-casting were replaced by the polygonal bulk-like α-Al8Fe2Si. As a result, the ultimate tensile strength and elongation were increased to 220 MPa and 2.6%, respectively.

Screening and Validation of a Carvacrol-Targeting Viability-Regulating Protein, SLC6A3, in Liver Hepatocellular Carcinoma.

Background: Liver hepatocellular carcinoma (LIHC) is the second leading cause of tumor-related death in the world. Carvacrol was also found to inhibit multiple cancer types. Here, we proposed that Carvacrol inhibited LIHC. Methods: We used MTT assay to determine the inhibition of Carvacrol on LIHC cells. BATMAN-TCM was used to predict targets of Carvacrol. These targets were further screened by their survival association and expression in cancer using TCGA data. The bioinformatic screened candidates were further validated in in vitro experiments and clinical samples. Finally, docking models of the interaction of Carvacrol and target protein were conducted. Results: Carvacrol inhibited the viability of LIHC cell lines. 40 target genes of Carvacrol were predicted, 8 of them associated with survival. 4 genes were found differentially expressed in LIHC vs. normal liver. Among these genes, the expression of SLC6A3 and SCN4A was found affected by Carvacrol in LIHC cells, but only SLC6A3 correlated with the viability inhibition of Carvacrol on LIHC cell lines. A docking model of the interaction of Carvacrol and SLC6A3 was established with a good binding affinity. SLC6A3 knockdown and expression revealed that SLC6A3 promoted the viability of LIHC cells. Conclusion: Carvacrol inhibited the viability of LIHC cells by downregulating SLC6A3.

Impact of spatial misallocation of electric power resources on economic efficiency and carbon emissions in China.

The relationship between resource misallocation and productivity has become a hot topic in recent years, but few studies examined the impact of spatial misallocation of electric power resources (SMEPRs) on economic efficiency and carbon emissions. Here, we constructed a calculation model of SMEPRs that can measure both the misallocation degree and direction and uncovered the spatiotemporal evolvement mechanism of SMEPRs. On this basis, we explored the impact of SMEPRs on regional economic efficiency and carbon emissions using panel data from 29 provinces in China from 1988 to 2017. The results demonstrate that the high level of SMEPRs in China shows complex spatiotemporal characteristics and significantly affects the regional economic efficiency and carbon emissions. Specifically speaking, first, SMEPRs present the characteristics of the coexistence of excessive and insufficient allocation among provinces and regions, the increasing extent of misallocation in the eastern and western regions, and the gradual decline in the central region; second, SMEPRs have a strong negative effect on the regional economic efficiency and carbon emissions by affecting regional industrial structures, which indicates that SMEPRs are an important factor restricting the high-quality development of regional economies. The research is conducive to the development of resource misallocation theory. Moreover, the research observations offer fresh insights to upgrading the high-quality and green development of China's power sector and promoting regional economic transformation and ecological sustainability.

Screening of the Key Genes for the Progression of Liver Cirrhosis to Hepatocellular Carcinoma Based on Bioinformatics.

Hepatocellular carcinoma (HCC), which is among the most globally prevalent cancers, is strongly associated with liver cirrhosis. Using a bioinformatics approach, we have identified and investigated the hub genes responsible for the progression of cirrhosis into HCC. We analyzed the Gene Expression Omnibus (GEO) microarray datasets, GSE25097 and GSE17549, to identify differentially expressed genes (DEGs) in these two conditions and also performed protein-protein interaction (PPI) network analysis. STRING database and Cytoscape software were used to analyze the modules and locate hub genes following which the connections between hub genes and the transition from cirrhosis to HCC, progression of HCC, and prognosis of HCC were investigated. We used the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to detect the molecular mechanisms underlying the action of the primary hub genes. In all, 239 DEGs were obtained, with 94 of them showing evidence of upregulation and 145 showing evidence of downregulation in HCC tissues as compared to cirrhotic liver tissues. We identified six hub genes, namely, BUB1B, NUSAP1, TTK, HMMR, CCNA2, and KIF2C, which were upregulated and had a high diagnostic value for HCC. Besides, these six hub genes were positively related to immune cell infiltration. Since these genes may play a direct role in the progression of cirrhosis to HCC, they can be considered as potential novel molecular indicators for the onset and development of HCC.

Glycolysis-related gene dihydrolipoamide acetyltransferase promotes poor prognosis in hepatocellular carcinoma through the Wnt/ß-catenin and PI3K/Akt signaling pathways.

Background: Recent research suggests that dihydrolipoamide acetyltransferase (DLAT), which is a copper-induced cell death-related gene, is involved in multiple biological events in tumors. This study sought to investigate the relationship between DLAT and hepatocellular carcinoma (HCC). Methods: In the Cancer Genome Atlas (TCGA) database, we first identified the differentially expressed gene (i.e., DLAT), then confirmed DLAT expression, and found a link between it and the prognosis of HCC patients. An internal validation nomogram was built based on a multivariate Cox regression analysis. Data from the Tumor Immune Estimation Resource (TIMER) database was used to examine the association between DLT and immunological cells. A gene set enrichment analysis (GSEA) was conducted to investigate the probable mechanism of action. Finally, in vitro cytological research was conducted to further examin the involvement of DLAT in HCC-related unfavorable biological events. Results: The database screenings showed that DLAT was a differentially expressed molecule; that is, DLAT was more highly expressed in the cancer tissues than normal tissues. TCGA results and Kaplan-Meier-plotter data sets showed that HCC patients with reduced DLAT expression had greater disease-specific survival (DSS), overall survival (OS), and progression-free interval (PFI). The prediction model had a concordance index of 0.659 (0.614-0.704), which indicates high accuracy. According to the TIMER database, tumor cells in the HCC microenvironment may be able to bypass the immune system due to the expression of DLAT. The in vitro cytological tests showed that DLAT knockdown significantly decreased the proliferation and invasion of the HCC cells. It also inhibited the activity of the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/protein kinase B (Akt) and Wnt/ß-catenin signaling pathways. Conclusions: Decreased DLAT expression significantly prolongs the OS, PFI, and DSS of HCC patients. DLAT may be employed as a new predictive biomarker for HCC, and may be linked to the immune system in HCC patients. The tumor microenvironment (TME) may have a significant effect on the ability of tumor cells to evade the immune system. The PI3K/Akt and Wnt/ß-catenin signaling pathways may affect the prognosis of HCC by interfering with DLAT. Given these findings, HCC may be an ideal target for the development of anti-cancer therapies.

Endogenous sulfur dioxide protects against isoproterenol-induced myocardial injury and increases myocardial antioxidant capacity in rats.

Recently, sulfur dioxide (SO(2)) was discovered to be produced in the cardiovascular system and to influence important biological processes. Here, we investigated changes in endogenous SO(2)/glutamic oxaloacetic transaminase (GOT) pathway in the development of isoproterenol (ISO)-induced myocardial injury in rats and the regulatory effect of SO(2) on cardiac function, myocardial micro- and ultrastructure, and oxidative stress. Wistar male rats were divided into control, ISO-treated, ISO+SO(2), and SO(2) groups. At the termination of the experiment, parameters of cardiac function and hemodynamics were measured and the micro- and ultrastructure of myocardium and stereological ultrastructure of mitochondria were analyzed. Myocardial SO(2) content was detected by high-performance liquid chromatography. GOT (key enzyme for endogenous SO(2) production) activity and gene (GOT1 and GOT2) expressions were measured, and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), hydrogen peroxide, and superoxide radical levels were assayed. SOD (SOD1 and SOD2) and GSH-Px (GSH-Px1) gene expressions were also detected. The results showed that SO(2) donor at a dose of 85 mg/(kg day) did not impact the cardiac function and structure of rats, but exerted a subtle influence on myocardial redox status. ISO-treated rats exhibited decreased cardiac function, damaged myocardial structures, and downregulated endogenous SO(2)/GOT pathway. Meanwhile, myocardial oxidative stress increased, whereas antioxidative capacity downregulated. Administration of SO(2) markedly improved cardiac function and ISO-induced myocardial damage by ameliorating the pathological structure of the myocardium and the mitochondria. At the same time, myocardial products of oxidative stress decreased, whereas antioxidative capacity increased. These results suggest that downregulation of the endogenous SO(2)/GOT pathway is likely involved in the pathogenesis of ISO-induced myocardial injury. SO(2) protects against ISO-induced myocardial injury associated with increased myocardial antioxidant capacity in rats.

Overexpression of the PLK4 Gene as a Novel Strategy for the Treatment of Autosomal Recessive Microcephaly by Improving Centrosomal Dysfunction.

Autosomal recessive microcephaly and chorioretinopathy (MCCRP) is a neurodevelopmental disorder characterized by delayed psychomotor development, growth retardation with dwarfism, and ocular abnormalities, and its occurrence has been found to be closely related to variants of the gene encoding centrosomes. However, the association between centrosomal duplication defects and the etiology of microcephaly syndromes is poorly understood. It is well known that polo-like kinase 4 (PLK4) is a key regulator of centriole duplication, and the abnormalities of centrosomal function caused by its protein variation need to be further explored in the pathogenesis of microcephaly. In our study, we found that a patient with microcephaly and chorioretinopathy harbored compound heterozygous missense variants NM_014264.4: c.2221C > T (p.Gln741*) and NM_014264.4: c.2062 T > C (p.Tyr688His) in the PLK4 gene. Overexpression experiments of the variant PLK4 proteins then showed that the G741 variant rather than the T688H variant had lost centrosomal amplification ability, and the G741 variant but not the T688H variant induced centrosomal replication disorder, which further inhibited cell proliferation, cycle division and cytoskeleton morphology in HeLa cells. Moreover, the overexpression of the two variant proteins had inconsistent effects on the target protein PLK4 by western blot analysis, also indicating that T688H variant overexpression is not functionally equivalent to WT-PLK4 overexpression. Therefore, all data support the idea that the PLK4 mutation induces centriolar duplication disorder and reduces the efficiency of mitosis inducing cell death or cell proliferation in the etiology of microcephaly disorder.

An Integrative Pan-Cancer Analysis of the Oncogenic Role of COPB2 in Human Tumors.

Several studies have suggested that coatomer protein complex subunit beta 2 (COPB2) may act as an oncogene in various cancer types. However, no systematic pan-cancer analysis has been performed to date. Therefore, the present study analyzed the potential oncogenic role of COPB2 using TCGA (The Cancer Genome Atlas) and GEO (Gene Expression Omnibus) datasets. The majority of the cancer types overexpressed the COPB2 protein, and its expression significantly correlated with tumor prognosis. In certain tumors, such as those found in breast and ovarian tissues, phosphorylated S859 exhibited high expression. It was found that mutations of the COPB2 protein in kidney and endometrial cancers exhibited a significant impact on patient prognosis. It is interesting to note that COPB2 expression correlated with the number of cancer-associated fibroblasts in certain tumors, such as cervical and endocervical cancers and colon adenocarcinomas. In addition, COPB2 was involved in the transport of substances and correlated with chemotherapy sensitivity. This is considered the first pan-tumor study, which provided a relatively comprehensive understanding of the mechanism by which COPB2 promotes cancer growth.

Exosomal microRNA-15a from mesenchymal stem cells impedes hepatocellular carcinoma progression via downregulation of SALL4.

Hepatocellular carcinoma (HCC) is a heterogeneous tumor with an increased incidence worldwide accompanied by high mortality and dismal prognosis. Emerging evidence indicates that mesenchymal stem cells (MSCs)-derived exosomes possess protective effects against various human diseases by transporting microRNAs (miRNAs or miRs). We aimed to explore the role of exosomal miR-15a derived from MSCs and its related mechanisms in HCC. Exosomes were isolated from transduced MSCs and co-incubated with Hep3B and Huh7 cells. miR-15a expression was examined by RT-qPCR in HCC cells, MSCs, and secreted exosomes. CCK-8, transwell, and flow cytometry were used to detect the effects of miR-15a or spalt-like transcription factor 4 (SALL4) on cell proliferative, migrating, invasive, and apoptotic properties. A dual-luciferase reporter gene assay was performed to validate the predicted targeting relationship of miR-15a with SALL4. Finally, in vivo experiments in nude mice were implemented to assess the impact of exosome-delivered miR-15a on HCC. The exosomes from MSCs restrained HCC cell proliferative, migrating, and invasive potentials, and accelerated their apoptosis. miR-15a was expressed at low levels in HCC cells and could bind to SALL4, thus curtailing the proliferative, migrating, and invasive abilities of HCC cells. Exosomes successfully delivered miR-15a to HCC cells. Exosomal miR-15a depressed tumorigenicity and metastasis of HCC tumors in vivo. Overall, exosomal miR-15a from MSCs can downregulate SALL4 expression and thereby retard HCC development.

Endovascular treatment or general treatment: how should acute ischemic stroke patients choose to benefit from them the most?: A systematic review and meta-analysis.

BACKGROUND: Acute ischemic stroke due to large-vessel occlusion is a leading cause of death and disability, and therapeutic time window was limited to 4.5 hour when treated with intravenous thrombolysis. It has been acknowledged that endovascular treatment (EVT) is superior to general treatment (only medication, including intravenous recombinant tissue plasminogen activator (rt-PA)) in improving the outcome of AIS since 2015. However, the benefits were limited to improvement of functional outcomes and functional independence. Hence, this meta-analysis was conducted to summarize the benefits of EVT for acute ischemic stroke, explore underlying indications of EVT for AIS patients and suggest implications for clinical practice and future research. METHODS: A search was performed to identify eligible studies in PubMed, Scopus and Web of Science updated to February 5, 2019. Functional outcomes, the modified Rankin Scale (mRS) 0-1, mRS 0-2, all-cause mortality, symptomatic intracerebral hemorrhage and asymptomatic intracerebral hemorrhage (aICH) at 90 days were selected as outcomes. Data was pooled to calculate the odds ratio (OR) and 95% confidence interval (CI). Heterogeneity, subgroup analysis, sensitivity analysis and publication bias were also performed in this meta-analysis. RESULTS: Eighteen studies comprising 3831 patients were included and analyzed in this meta-analysis. In comparison with general treatment, improved functional outcomes (mRS 0-1: OR = 1.68, 95% CI = 1.43-1.97, inconsistency index [I = 57%, P < .00001; mRS 0-2: OR = 1.78, 95% CI = 1.55-2.03, I = 69%, P < .00001), reduced risk of all-cause mortality (OR = 0.82, 95% CI = 0.70-0.98, I = 27%, P = .03) but higher risk of aICH (OR = 1.43, 95% CI = 1.05-1.95, I = 0%, P = .02) at 90 days were found in AIS patients treated with EVT. Age < 70, National Institutes of Health Stroke Scale ≥20 and maximum delay for invention>5 hours could improve clinical outcomes following EVT. In sensitivity analysis, it showed that 2 studies had a great influence on the pooled ORs. No potential publication bias was found in this meta-analysis. CONCLUSION: Taken together, EVT, which led to improved functional outcomes and decreased risk of death, is superior to general treatment for AIS patients with age < 70, National Institutes of Health Stroke Scale ≥20 and maximum delay for invention>5 hours. Moreover, it suggests that "with mechanical thrombectomy" is potential favorable factor for improving aICH in comparison with general treatment.

Whole-Transcriptome Sequencing Identifies Key Differentially Expressed mRNAs, miRNAs, lncRNAs, and circRNAs Associated with CHOL.

To systematically evaluate the whole-transcriptome sequencing data of cholangiocarcinoma (CHOL) to gain more insights into the transcriptomic landscape and molecular mechanism of this cancer, we performed whole-transcriptome sequencing based on the tumorous (C) and their corresponding non-tumorous adjacent to the tumors (CP) from eight CHOL patients. Subsequently, differential expression analysis was performed on the C and CP groups, followed by functional interaction prediction analysis to investigate gene-regulatory circuits in CHOL. In addition, The Cancer Genome Atlas (TCGA) for CHOL data was used to validate the results. In total, 2,895 differentially expressed messenger RNAs (dif-mRNAs), 56 differentially expressed microRNAs (dif-miRNAs), 151 differentially expressed long non-coding RNAs (dif-lncRNAs), and 110 differentially expressed circular RNAs (dif-circRNAs) were found in CHOL samples compared with controls. Enrichment analysis on those differentially expressed genes (DEGs) related to miRNA, lncRNA, and circRNA also identified the function of spliceosome. The downregulated hsa-miR-144-3p were significantly enriched in the competing endogenous RNA (ceRNA) complex network, which also included 7 upregulated and 13 downregulated circRNAs, 7 upregulated lncRNAs, and 90 upregulated and 40 downregulated mRNAs. Moreover, most of the DEGs and a few of the miRNAs (such as hsa-miR-144-3p) were successfully validated by TCGA data. The genes involved in RNA splicing and protein degradation processes and miR-144-3p may play fundamental roles in the pathogenesis of CHOL.

[Hydrogen sulfide induce negative inotropic effect in isolated hearts via KATP channel and mitochondria membrane KATP channel].

OBJECTIVE: Hydrogen sulfide (H(2)S) dilates blood vessels in vivo and in vitro probably by opening vascular smooth muscle K(+)-ATP channels. The study was designed to observe the role of mitochondria membrane K(ATP) channel blocker (5-HD) in the regulation of cardiac function isolated perfused heart of rat with H(2)S. METHODS: The isolated rat heart was perfused in a Langendorff apparatus. After 20 minutes of stabilization, physiological concentration of NaHS (H(2)S donor, 100 micromol/L) was continuously perfused for 20 min in group A (n = 6), isolated hearts in group B (n = 6) and C (n = 7) were pretreated with nonspecific K(ATP) channel blocker glibenclamide (100 micromol/L) or 5-HD (100 micromol/L) for 5 minutes then perfused with NaHS (100 micromol/L) for 10 minutes. Heart rate (HR), left ventricular developed pressure (DeltaLVP), dp/dt(max) and dp/dt(min) and coronary perfusion flow (CPF) were measured. RESULTS: Post continuous perfusion of NaHS at physiological concentration for 20 minutes, DeltaLVP, dp/dt(max) and dp/dt(min) all significantly decreased while HR and CPF remained unchanged compared to baseline levels (all P < 0.05). The negative inotropic effect of H(2)S could partly be blocked by nonspecific K(ATP) channel blocker glibenclamide and mitochondria membrane K(ATP) channel blocker 5-HD. CONCLUSION: Present findings suggested that H(2)S at physiological concentration could produce negative inotropic effect in isolated hearts and this effect was mediated by K(ATP) channel and mitochondria membrane K(ATP) channel.

Synthesis of Nanocrystalline AZ91 Magnesium Alloy Dispersed with 15 vol.% Submicron SiC Particles by Mechanical Milling.

The development of a magnesium matrix composite with a high content of dispersions using conventional liquid-phase process is a great challenge, especially for nanometer/submicron particles. In this work, mechanical milling was employed to prepare nanocrystalline AZ91 dispersed with 15 vol.% submicron SiC particles (SiCp/AZ91). AZ91 with no SiCp was applied as a comparative study with the same mechanical milling. In order to investigate the mechanism of dispersing, the morphology evolution of powders and the corresponding SiCp distribution were observed. As the scanning electron microscope (SEM) analysis exhibited, the addition of SiCp accelerated the smashing of AZ91 particles, which promoted the dispersion of SiCp in AZ91. Thus, after mechanical milling, 15 vol.% SiCp, which was smashed from 800 to 255 nm, got uniformly distributed in the Mg matrix. Based on X-ray diffraction (XRD) results, part of the Mg17Al12 precipitate got dissolved, and an Al-supersaturated Mg solid solution was formed. The transmission electron microscopy (TEM) results showed that the ultimate Mg grain (32 nm) of milled SiCp/AZ91 was much smaller than that of milled AZ91 (64 nm), which can be attributed to a pinning effect of submicron SiCp. After mechanical milling, the hardness of SiCp/AZ91 reached 185 HV, which was 185% higher than the original AZ91 and 33% higher than milled AZ91, due to fine Mg grain and submicron dispersions.

[Effects of auto-tumor infiltrating lymphocytes induced by interleukin (IL)-12 with IL-2 on patients of primary hepatic carcinoma].

OBJECTIVE: To investigate the effects of the tumor infiltrating lymphocytes (TILs) from primary hepatic carcinoma (PHC) induced by interleukin-12 (IL-12) with IL-2, on their cytotoxicity, proliferation, and cytokine production, and the immunological function and survival of the PHC patients. METHODS: PHC cells and TILs were isolated from the resected tumors and cultured. IL-2 was added into the culture medium with or without IL-12. Ten to 14 days later the cytotoxic activity and proliferation index (PI) of the TILs were calculated. The levels of the cytokine interferon (IFR)-gamma and tumor necrosis factor (TNF)-alpha in the supernatant were tested by ELISA. 20-25 days after the operation the TILs were re-infused into the bodies. The clinical manifestation, cytotoxicity of TILs, phenotype of peripheral blood lymphocytes, and survival of patients were observed. RESULTS: 1. The cytotoxicity of the TILs of the IL-12 + IL-2 group against the autologous hepatic carcinoma cells was greater than that of the TILs of the IL-2 group (P < 0.05). 2. The PI of the TILs of the IL-12 + IL-2 group was 17.78%, significantly higher than that of the IL-2 group (10.9%, P < 0. 05). 3. The levels of IFN-gamma and TNF-alpha of the IL-12 + IL-2 group were (2180 +/- 494) pg/ml and(485 +/- 98) pg/ml, both significantly higher than those of the IL-2 group [(1078 +/- 309.46) pg/ml and (422.00 +/- 15.81) pg/ml, both P < 0. 05]. 4. After re-infusion of TILs, the CD3+, CD8+, CD4+, and CD56+ rates, especially the CD3+ and CD8+ rates, of the IL-12 + IL-2 group were all significantly higher than those of the IL-2 group (all P < 0.05). 5. After TIL re-infusion the clinical manifestation of all patients were improved. 6. 23 of the 25 patients receiving the re-infusion of the TILs of the IL-12 + IL-2 group survived for more than 1 year, with a rate significantly higher than that of the IL-2 group (17/25, chi2 = 4.5, P < 0.05), however, there were no significant differences in the 3-year and 5-year survival rates between these 2 groups (both P > 0.05). The number of patients who showed recurrence in less than 1 year was 17 in the IL-2 group, with a rate significantly higher that of the IL-12 + IL-2 group (8/25, chi2 = 4.32, P < 0.0 5). However, there was no significant difference in the recurrence rate < or = 3 years between these 2 groups (chi2 = 1.56, P > 0.05). CONCLUSION: TILs from primary hepatic carcinoma induced by IL-12 can obviously enhance specific cytotoxicity and proliferation of TILs. TILs induced by IL-12 + IL-2 increase the patients' immunological function, prolong the 1-year survival, and decreases recurrence more effectively than the TILs induced by IL-2 only.

[Effects of sulfur dioxide on cardiac function of isolated perfusion heart of rat].

OBJECTIVE: To observe the effects of sulfur dioxide (SO2) on cardiac function of isolated perfused heart of rat and to explore the physiological regulation of endogenous SO2 on myocardial action. METHODS: The hearts of 64 Wistar rats were isolated, perfused with Krebs-Henseleit (KH) solution through Langendorff apparatus, and randomly divided into 8 equal groups: Four groups underwent perfusion of SO2 of the concentrations 1, 10, 100, 1000 micromol/L respectively for 5 min and then perfused with KH solution for 15 min. Eight hearts underwent perfusion of SO2 of the physiological concentration (10 micromol/L) for 20 min. The control group underwent perfusion of KH solution for 20 min. Eight hearts of the nicardipine group underwent perfusion of nicardipine, a L-type calcium channel blocker, 2.5 micromol/L for 5 min, SO2 10 micromol/L for 5 min, and then KH solution for 10 min. The heart in the hydroxamate (HDX) group underwent perfusion of HDX, an inhibitor of SO2 endogenous generating enzymes, for 5 min, and then perfused by KH solution for 15 min. The heart rate (HR), difference of left ventricular pressure (LVP), left ventricular peak rate of contraction (+ dp/dtmax), peak rate of relaxation (- dp/dtmax), and coronary flow (CF) were measured. Then transmission electron microscopy was conducted. RESULTS: SO2 concentration-dependently inhibited the left ventricular +/- dp/dtmax, LVP, HR, and CF (all P < 0.01). The left ventricular +/- dp/dtmax, LVP, and HR were inhibited (P < 0.05) by the physiological concentration (10 micromol/L) SO2 donor continuous perfusion for 20 min. During perfusion 20 min, the LVP, + dp/dtmax, - dp/dtmax, and HR after perfusion for 20 min of the physiological concentration (10 micromol/L) SO2 donor continuous perfusion group were (15 +/- 3) mm Hg, (485 +/- 74) mm Hg/s, (339 +/- 64) mm Hg/s, and (114 +/- 26)/min respectively, all significantly lower than those 5 min after perfusion [(23 +/- 7) mm Hg, (595 +/- 93)mm Hg/s, (436 +/- 83) mm Hg/s, and (159 +/- 31)/min, all P < 0.05]. The LVP, + dp/ dtmax, -dp/dtmax, and HR of the nicardipine group were(37 +/- l0)mm Hg, (1025 +/- 287)mm Hg/s, (570 +/- 181)mm Hg/s, and (139 +/- 48)/min respectively, all not significantly different from those of the control group. The LVP, + dp/dtmax, - dp/dtmax, and CF after perfusion of the HDX group were (50 +/- 11)mm Hg, (1167 +/- 270) mm Hg/s, (889 +/- 72) mm Hg/s, and (6.3 +/- 1.9) ml/min respectively, all significantly lower than those before perfusion [(69 +/- 16) mm Hg, (1579 +/- 315) mm Hg/s, (1186 +/- 263) mm Hg/s, and (9.5 +/- 1.3) ml/min, all P < 0.05]. CONCLUSION: Exogenous SO2 has negative inotropic effect on myocardium. by the mechanism related to voltage-gated calcium channel. Nicardipine blocks the inhibitory effect of SO2 at physiological concentration.

Clinicopathological significance of cytoplasmic transducer of ErbB2. 1 expression in gastric cancer.

The aim of the present study was to investigate the expression of transducer of ErbB2. 1 (TOB1) in gastric carcinoma and to clarify the association between TOB1 expression and the clinical significance of this expression in patients with gastric carcinoma. Western blot analysis was performed to confirm the expression of TOB1 in gastric cancer. Immunohistochemistry (IHC) was performed on a tissue microarray containing 90 pairs of primary gastric cancer and adjacent normal tissue samples. TOB1 expression was evaluated separately with cytoplasmic and nuclear staining. Western blot analysis revealed significantly lower expression levels of TOB1 in gastric cancer tissues than those in adjacent normal tissues in 91.7% of cases. This was confirmed by IHC, which revealed decreased cytoplasmic TOB1 expression in cancer tissues compared with those of normal tissue samples in 84.4% of cases. The IHC data also revealed low cytoplasmic expression of TOB1 in 67.8% of human gastric cancer samples. Nuclear TOB1 expression exhibited no significant association with specific pathological features. However, a significant association was identified between cytoplasmic expression levels of TOB1 and clinicopathological characteristics, including the depth of invasion (P=0.017), differentiation grade (P=0.034) and tumor-node-metastasis stage (P<0.000). In conclusion, cytoplasmic TOB1 expression was suggested to be significant in angiogenesis and cell differentiation in gastric cancer tissues and may be used as a potential prognostic marker.