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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., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

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.

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.

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.

Since the 1980's nitric oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H(2)S), the endogenous gas molecules produced from metabolic pathway, have been realized as signal molecules to be involved in the regulation of body homeostasis and to play important roles under physiological and pathophysiological conditions. The researches on these endogenous gas signal molecules opened a new avenue in life science. To explore the new member of gasotransmitter family, other endogenous gas molecules which have been regarded as metabolic waste up to date, and their biological regulatory effects have been paid close attention to in the current fields of life science and medicine. Sulfur dioxide (SO(2)) can be produced endogenously from normal metabolism of sulfur-containing amino acids. L-cysteine is oxidized via cysteine dioxygenase to L-cysteinesulfinate, and the latter can proceed through transamination by glutamate oxaloacetate transaminase (GOT) to beta-sulfinyl pyruvate which decomposes spontaneously to pyruvate and SO(2). In mammals, activated neutrophils by oxidative stress can convert H(2)S to sulfite through a reduced form of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase-dependent process. The authors detected endogenous production of SO(2) in all cardiovascular tissues, including in heart, aorta, pulmonary artery, mesenteric artery, renal artery, tail artery and the plasma SO(2) content. As the key enzyme producing SO(2), GOT mRNA in cardiovascular system was detected and found to be located enriched in endothelial cells and vascular smooth muscle cells near the endothelial layer. When the normal rats were treated with hydroxamate(HDX), a GOT inhibitor, at a dose of 3.7 mg/kg body weight, the blood pressure (BP) went high markedly, the ratio of wall thickness to lumen radius was increased by 18.34%, and smooth muscle cell proliferation was enhanced. The plasma SO(2) level in the rats injected with 125 micromol/kg body weight SO(2) donor was increased to 721.98+/-30.11 micromol/L at the end of 30 seconds, while the blood pressure was decreased to the lowest point 65.0+/- 4.9 mm Hg at the end of 1 minute. The above results showed that endogenous SO(2) might be involved in the maintenance of blood pressure and normal vascular structure. In spontaneous hypertensive rat (SHR) animal model, exogenous supplement of SO(2) donor decreased the BP, the media cross-sectional area, and pressure of the media and the ratio of wall thickness to lumen radius in the SHR. Moreover, the proliferative index of aortic smooth muscle cells was decreased in the SHR treated with SO(2) donor compared with that in SHR. The above data showed that SO(2) could prevent the aortic structural remodeling by inhibiting the proliferation of aortic smooth muscle cells. The authors observed the direct vasorelaxant effects of SO(2) on the aortic ring pre-treated with norepinephrine (NE). SO(2) donor at a concentration of 25-100 micromol/L relaxed the aortic ring temporarily and slightly, but SO(2) donor at a concentration of 1-12 mmol/L induced relaxation of the ring in a concentration-dependent manner. Administration with nicardipine, an L-type calcium channel blocker other than glibenclamide, an ATP sensitive potassium channel (K(ATP) channel) blocker or removal of vascular endothelium could decrease the SO(2)-induced vasorelaxation. In hypoxic pulmonary hypertension animal model, SO(2) donor decreased the mean pulmonary artery pressure and the systolic pulmonary artery pressure (P<0.01), respectively as compared with hypoxic group, and alleviated obviously the hypoxic pulmonary vascular structural remodeling. The percentage of muscularized arteries of small pulmonary vessels was significantly decreased in hypoxia+SO(2) donor-treated rats compared with that of hypoxic rats (P<0.01), while the percentage of non-muscularized vessels was obviously higher in hypoxia with SO(2) donor-treated rats than that of hypoxic rats (P<0.01). Similarly, SO(2) obviously decreased relative media area and relative media thickness of small muscularized pulmonary arteries in hypoxic rats (P<0.01). The above data showed that SO(2) might play an important role in development of hypoxic pulmonary hypertension. Perfusion with SO(2) donor (10(-6)-10(-3) mol/L) to the isolated rat heart obviously inhibited the left ventricular peak rate of contraction ( + LV dp/ dtmax) , peak rate of relaxation (-LV dp/ dtmax) and difference of left ventricular pressure ( DeltaLVP) in a concentration dependent manner. Nicardipine, an L-type calcium channel blocker, could partly antagonize the inhibitory effect of SO(2) on the heart function. In a word, SO(2) could be endogenously generated in cardiovascular tissues and exert important cardiovascular effects such as vasorelaxant effect and negative inotropic effects. Moreover, SO(2) might play considerable roles in the regulation of systemic circulatory pressure, pulmonary circulatory pressure and vascular structural remodeling in the pathogenesis of hypertension and hypoxic pulmonary hypertension. On the basis of the above findings, we presumed that endogenous SO(2) might be a novel cardiovascular functional regulatory gasotransmitter. More studies on the significance of endogenous SO(2) in cardiovascular system under physiological and pathophysiological conditions need to be investigated.

Objective: To study the effect of safflower injection (SI) in protecting heart, and on energy charge and anti-apoptosis gen bcl-2 in cardiac tissue., Methods: Rats' Langendorff isolated heart infused model was used in the experiment to study the effect of SI by measuring the cardiac function, energy charge and bcl-2 expression of the cultured heart in the modified Euro-Collins (mEC) heart preserving liquid with or without addition of SI., Results: As compared with the control, SI showed the effects of improving functions of cardiac contraction and dilation, increasing coronary blood flow, and strengthening the bcl-2 protein expression., Conclusion: SI has excellent effect in protecting heart.

Objective: To observe the effect of Jisheng injection (JSI) in protecting heart., Methods: Isolated heart of rat was preserved in modified Euro-Collins solution (mEC) containing JSI for 20 hrs, and that preserved in simple mEC was taken as control. Then Langendorff isolated rat heart perfusion was conducted. Forty minutes after perfusion, the cardiac function, coronary flow, myocardial water content were determined, and lactate dehydrogenase (LDH), creatine kinase (CK) activity in perfusate, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content in myocardial tissue and pathologic change in myocardium were also observed., Results: The cardiac function and coronary flow of isolated heart preserved in JSI containing mEC was significantly better than those in the control (P < 0.01), with the LDH, CK activity and MDA content significantly lower (P < 0.01 and P < 0.05), SOD activity significantly higher (P < 0.05) and pathologic injury milder than those in control, but comparison of cardiac water content between the two groups showed insignificant difference., Conclusion: JSI has good cardiac protective and anti-oxidizing effects.