Objective: To observe the dynamic changes of plasma level thymosinβ4 (Tβ4) in acute myocardial infarction (AMI) patients with intervening therapy within 15 days of onset and to explore the relationship between Tβ4 and clinical prognosis in AMI patients.
Methods: Our research included 2 groups:AMI group, n=69 and Control group, the patients with suspected chest pain while CAG excluded coronary artery stenosis, n=32. Plasma levels of Tβ4 were examined in all AMI patients on admission day and every day until 15 days of onset;AMI patients were followed-up for 18 months and the endpoint was defined as major adverse cardiovascular event (MACE) occurrence.
Results: ①Compared with Control group, AMI group had increased plasma level of Tβ4 on admission day and on day-15 of onset, P<0.01. ② With intervening therapy, AMI group had elevated Tβ4 level upon immediate onset, it was decreased on day-1, reached low level on day-3 and elevated to peak on day-6, then reduced followed by slightly raising on day-11.③During follow-up period, the AMI patients without MACE had the higher mean in-hospital maximum Tβ4 value than those with MACE occurrence, P<0.01. Logistic regression analysis indicated that the mean in-hospital maximum Tβ4 value was related to MACE occurrence during follow-up period (OR=0.999, 95%CI 0.999-1.000).
Conclusion: AMI may induce up-regulated expression of plasma Tβ4;with intervening therapy, Tβ4 showed a trend of“elevation-reduction-elevation-reduction”at the early stage of AMI. High expression of Tβ4 was helpful for improving clinical prognosis in AMI patients which may provide a theoretical basis for exogenous use of Tβ4 in AMI treatment.

Objective:To evaluate the role of signal transducer and activator of transcription 3 (STAT3)/nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in salidroside-induced attenuation of intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Thirty-six SPF-grade healthy male C57BL mice, aged 6-8 weeks, weighing 20-25 g, were divided into 6 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation+ salidroside group (SS group), intestinal I/R group (IR group), intestinal I/R+ salidroside group (IS group), intestinal I/R+ salidroside+ autophagy activator rapamycin group (ISR group) and intestinal I/R+ salidroside+ STAT3 activator colivelin group (ISC group). The intestinal I/R injury model was established by clamping the superior mesenteric artery for 45 min followed by 30-min reperfusion in IR, IS, ISR and ISC groups, while the superior mesenteric artery was only isolated without clipping in S and SS groups. At 1 week before developing the model, salidroside 40 mg/kg was intraperitoneally injected once a day for 7 consecutive days in SS, IS, ISC and ISR groups, rapamycin 4 mg/kg was intraperitoneally injected once a day for 7 consecutive days in group ISR, colivelin 1 mg/kg was intraperitoneally injected once a day for 7 consecutive days in group ISC, while the equal volume of normal saline was given instead in the rest two groups. The mice were sacrificed at 30 min of reperfusion, and intestinal tissues were obtained for examination of the pathological changes after HE staining (with a optical microscope) which were scored according to Chiu and for determination of contents of malondialdehyde (MDA), Fe 2+, glutathione (GSH) and reactive oxygen species (ROS), activity of superoxide dismutase (SOD) and expression of p-STAT3, STAT3, glutathione peroxidase 4 (GPX4), NCOA4 and ferritin heavy chain 1 (FTH1) in intestinal tissues (by Western blot). Results:Compared with group S, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly increased, the content of GSH was decreased, the activity of SOD was decreased, the expression of p-STAT3 and NCOA4 was up-regulated, the expression of GPX4 and FTH1 was down-regulated, the p-STAT3/STAT3 ratio was increased ( P<0.05), pathological injury was found in intestinal tissues, and no significant change was found in the aforementioned indexes in group IR( P>0.05). Compared with group IR, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly decreased, GSH content was increased, SOD activity was increased, the expression of p-STAT3 and NCOA4 was down-regulated, the expression of GPX4 and FTH1 was up-regulated, p-STAT3/STAT3 ratio was decreased ( P<0.05), and the pathological injury was significantly alleviated in intestinal tissues in group IS. Compared with group IS, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly increased, GSH content was decreased, SOD activity was decreased, the expression of p-STAT3 and NCOA4 was up-regulated, the expression of GPX4 and FTH1 was down-regulated, p-STAT3/STAT3 ratio was increased ( P<0.05), and the pathological injury was aggravated in intestinal tissues in ISR and ISC groups. There was no statistically significant difference in the expression of STAT3 among the five groups ( P>0.05). Conclusions:STAT3/NCOA4-mediated ferritinophagy is involved in the process of salidroside-induced reduction of intestinal I/R injury in mice, which may be related to inhibiting ferroptosis.

Objective:To evaluate the role of endoplasmic reticulum stress-mediated apoptosis in proanthocyanidins-induced attenuation of intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty SPF healthy adult male C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 5 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation + proanthocyanidins group (S+ PC group), intestinal I/R group (I/R group), intestinal I/R + proanthocyanidins group (I/R+ PC group) and intestinal I/R + proanthocyanidins + tunicamycin group (I/R+ PC+ TM group). The superior mesenteric artery was clamped for 60 min and reperfused for 120 min to establish a mouse intestinal I/R injury model.Proanthocyanidin 100 mg/kg was given by intragastric gavage every day 1 week before ischemia in S+ PC group, I/R+ PC group and I/R+ PC+ TM group, and the equal volume of normal saline was given for 7 consecutive days in S group and I/R group, and endoplasmic reticulum stress agonist tunicamycin 1 mg/kg was intraperitoneally injected at 24 h before ischemia in I/R+ PC+ TM group.The mice were sacrificed at 120 min of reperfusion, and the small intestinal tissues were taken for microscopic examination of the histopathological changes (using light microscopy) and for determination of the level of diamine oxidase (DAO) (by enzyme-linked immunosorbent assay), cell apoptosis (by TUNEL method), glucose regulatory protein 78 (GRP78), C/EBP-homologous protein (CHOP) and cleaved caspase-3, Bax and Bcl-2 (by Western blot). Intestinal damage was assessed and scored according to Chiu, and the apoptosis index (AI) and Bcl-2/Bax ratio were calculated. Results:Compared with S group, the Chiu′s score, level of DAO and AI were significantly increased, the expression of GRP78, CHOP, cleaved caspase-3 and Bax was up-regulated, the expression of Bcl-2 was down-regulated, the ratio of Bcl-2/Bax was decreased( P<0.05), and the pathological damage occurred in the small intestinal tissue in I/R group, and no significant change was found in the aforementioned indexes in S+ PC group ( P>0.05). Compared with I/R group, the Chiu′s score, DAO level and AI were significantly decreased, the expression of GRP78, CHOP, cleaved caspase-3 and Bax was down-regulated, the expression of Bcl-2 was up-regulated, the ratio of Bcl-2/Bax was increased( P<0.05), and the pathological injury to the small intestinal tissue was significantly reduced in I/R+ PC group. Compared with I/R+ PC group, the Chiu′s score, level of DAO and AI were significantly increased, the expression of GRP78, CHOP, cleaved caspase-3 and Bax was up-regulated, the expression of Bcl-2 was down-regulated, and the ratio of Bcl-2/Bax was decreased( P<0.05), and the pathological damage to the small intestinal tissue was aggravated in I/R+ PC+ TM group. Conclusions:Proanthocyanidins can alleviate intestinal I/R injury by inhibiting endoplasmic reticulum stress-mediated cell apoptosis in mice.

Objective:To evaluate the role of silencing information regulatory factor 1/nuclear factor E2 related factor 2 (SIRT1/Nrf2) signaling pathway in berberine preconditioning-induced reduction of intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Thirty-six SPF-grade healthy male C57BL/6 mice, aged 8-10 weeks, weighing 22-25 g, were divided into 6 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation + berberine preconditioning group (SB group), intestinal I/R group (IR group), intestinal I/R + berberine preconditioning group (B group), intestinal I/R + berberine preconditioning + SIRT1 inhibitor EX527 group (BE group) and berberine preconditioning + intestinal I/R + ferroptosis inducer RSL3 group (BR group). The model of intestinal I/R injury was prepared by clamping the superior mesenteric artery for 45 min followed by 30-min reperfusion in IR group, B group, BE group and BR group, while the superior mesenteric artery was only isolated without ligation in S group and SB group. Berberine 50 mg/kg was administered by intragastric gavage once a day starting from 7 days before developing the model in SB group, B group, BE group and BR group. EX527 5 mg/kg and RSL3 5 mg/kg were intraperitoneally injected once a day at 3 days before surgery in BE group and BR group, respectively. The equal volume of normal saline was given in the other groups. The mice were sacrificed at 30 min of reperfusion, and the intestinal tissues were taken for microscopic examination of the pathological changes of intestinal mucosa (with a light microscope) which was scored according to Chiu and for determination of the contents of Fe 2+ and malondialdehyde (MDA) and superoxide dismutase (SOD) activity (by colorimetry), glutathione (GSH) content (by enzyme-linked immunosorbent assay), reactive oxygen species (ROS) content (by fluorescence staining), and expression of glutathione peroxidase 4 (GPX4), SIRT1 and Nrf2 (by Western blot). Results:Compared with S group, Chiu′s score was significantly increased, the contents of Fe 2+, MDA and ROS were increased, the content of GSH and activity of SOD were decreased, the expression of GPX4 was down-regulated, and the expression of SIRT1 and Nrf2 was up-regulated in IR group ( P<0.05), and no significant change was found in Chiu′s score in SB group ( P>0.05). Compared with IR group, Chiu′s score was significantly decreased, the contents of Fe 2+, MDA and ROS were decreased, the content of GSH and activity of SOD were increased, the expression of GPX4 was up-regulated, and the expression of SIRT1 and Nrf2 was up-regulated in B group ( P<0.05). Compared with B group, Chiu′s score was significantly increased, the contents of Fe 2+, MDA and ROS were increased, the content of GSH and activity of SOD were decreased, and the expression of GPX4 was down-regulated in BE and BR groups, and the expression of SIRT1 and Nrf2 was down-regulated in BE group( P<0.05). Conclusions:The mechanism by which berberine preconditioning reduces intestinal I/R injury may be associated with activation of SIRT1/Nrf2 signaling pathway, thus inhibiting ferroptosis in mice.

AIM: To screen the active constituents in Taohong Siwu Decoction(THSWD) by Systems Pharmacology and to study its protective effect on myocardial injury through cell experiments. METHODS: The TCMSP database and Pharmmaper database were used to retrieve the constituents and active constituent targets of THSWD. The Genecards database was used to retrieve the myocardial injury related targets gene. Then construct and analyze the constituents-disease intersection target network and the contribution value of constituents, and screen out the main active constituents of THSWD acting on the myocardium. Furthermore, CoCl

Objective To explore the effects of astaxanthin on pressure injury wounds.Methods In vitro experiment:Fibroblasts were treated with different concentrations of astaxanthin and their proliferation activity was detected by CCK-8 assay.Subsequently,fibroblasts were induced by hypoxia/reoxygenation,and the optimal concentration of astaxanthin was administered.Then the intracellular reactive oxygen species(ROS)level was detected by DHE fluorescent probes and the mRNA expression level of TNF-α,IL-1β,IL-6,IL-10,TGF-β was evaluated by RT qPCR.In vivo experiment:to construct a pressure injury model,two circular magnets were symmetrically adsorbed on both sides of the mouse skin for 5 hours everyday.Subsequently,equal amounts of physiological saline,low-dose astaxanthin(10 mg/kg),and high-dose astaxanthin(20 mg/kg)were administered by gavage in groups.Wound images were taken regularly.After 7-day treatment,wound healing rates were counted and wound tissues were collected for histopathological staining.Results In vitro,the fluorescence intensity of DHE in the astaxanthin groups were reduced dramatically.The relative mRNA expression level of TNF-α,IL-1β,IL-6 in the astaxanthin group declined,and the level of TGF-β and IL-10 mRNA increased significantly(P<0.05).In vivo,the wound healing rate and the level of TGF-β,IL-10 in high-dose astaxanthin group increased significantly.The ROS content and the level of TNF-α,IL-1β and IL-6 dropped markedly in astaxanthin groups(P<0.05).Conclusions Astaxanthin can significanlty alleviate oxidative stress,mitigate inflammation,thus exerting a protective effect on pressure injury wounds.

Organic anion transporting polypeptide 1B1 and 1B3 (OATP1B1/3) as important uptake transporters play a fundamental role in the transportation of exogenous drugs and endogenous substances into cells. Rat OATP1B2, encoded by the gene, is homologous to human OATP1B1/3. Although OATP1B1/3 is very important, few animal models can be used to study its properties. In this report, we successfully constructed the S knockout (KO) rat model using the CRISPR/Cas9 technology for the first time. The novel rat model showed the absence of OATP1B2 protein expression, with no off-target effects as well as compensatory regulation of other transporters. Further pharmacokinetic study of pitavastatin, a typical substrate of OATP1B2, confirmed the OATP1B2 function was absent. Since bilirubin and bile acids are the substrates of OATP1B2, the contents of total bilirubin, direct bilirubin, indirect bilirubin, and total bile acids in serum are significantly higher in KO rats than the data of wild-type rats. These results are consistent with the symptoms caused by the absence of OATP1B1/3 in Rotor syndrome. Therefore, this rat model is not only a powerful tool for the study of OATP1B2-mediated drug transportation, but also a good disease model to study hyperbilirubinemia-related diseases.