Objective:To evaluate the role of exosomes in propofol-induced elimination of cardioprotective effect of remote ischemic preconditioning (RIPC) in rats.Methods:This experiment was performed in 2 parts. In vivo experiment Forty-eight healthy SPF male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 5 groups using the random number table method: sham operation group (Sham group, n=12), ischemia-reperfusion (I/R) group ( n=12), RIPC group ( n=8), RIPC+ propofol group (RIPC+ P group, n=8), and propofol+ I/R group (P+ I/R group, n=8). The model of myocardial I/R injury was developed by ligating the left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion in anesthetized animals. Four cycles of 5-min ischemia induced by occlusion of the bilateral hind limbs with a tourniquet/5-min reperfusion served as the RIPC stimulus. Propofol was intravenously infused at a rate of 12 mg·kg -1·h -1 in RIPC+ P group (during RIPC) and in P+ I/R group (for 40 min). Exosomes from RIPC-treated and RIPC+ propofol-treated rats were extracted (RIPC-EXO and RIPC+ P-EXO respectively) for determination of the expression of surface markers of exosomes CD9 and HSP70. Another 24 rats were randomly selected, and the aforementioned exosomes were injected at 15 min before myocardial ischemia, resulting in RIPC-EXO+ I/R group ( n=12) and RIPC+ P-EXO+ I/R group ( n=12). At the end of reperfusion, the area of myocardial infarction was determined, the concentration of serum cardiac troponin I (cTnI) was measured by the enzyme-linked immunosorbent assay, and the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in myocardial tissues was detected by Western blot. Cell experiment H9c2 cells were cultured in vitro and divided into 4 groups ( n=6 each) using a random number table method: control group (C group), hypoxia-reoxygenation group (H/R group), RIPC-EXOc group and RIPC+ P-EXOc group. The cells were exposed to hypoxia for 4 h followed by reoxygenation for 16 h in H/R group. RIPC-EXO and RIPC+ P-EXO were added at a final concentration of 300 μg/ml before hypoxia in RIPC-EXOc group and RIPC+ P-EXOc group, respectively. The cell viability was determined using a cell counting kit-8 assay and the expression of Bax and Bcl-2 expression was detected by Western blot. Results:In vivo experiment Compared with RIPC-EXO group, the expression of CD9 and HSP70 was significantly down-regulated in RIPC+ P-EXO group ( P<0.05). Compared with Sham group, the percentage of the area of myocardial infarction was significantly increased, and the serum cTnI concentration and Bax/Bcl-2 ratio in myocardial tissues were increased in I/R group ( P<0.05). Compared with I/R group, the percentage of the area of myocardial infarction was significantly decreased in RIPC group and RIPC-EXO+ I/R group, the serum cTnI concentration and Bax/Bcl-2 ratio in myocardial tissues were significantly decreased in RIPC-EXO+ I/R group ( P<0.05), and no significant change was found in the percentage of the area of myocardial infarction in RIPC+ P group ( P>0.05). The percentage of the area of myocardial infarction was significantly larger in RIPC+ P group than in RIPC group ( P<0.05). Compared with RIPC-EXO+ I/R group, the percentage of the area of myocardial infarction was significantly increased, and the serum cTnI concentration and Bax/Bcl-2 ratio were increased in RIPC+ P-EXO+ I/R group ( P<0.05). Cell experiment Compared with C group, the cell viability was significantly decreased, and the Bax/Bcl-2 ratio was increased in H/R group ( P<0.05). Compared with H/R group, the cell viability was significantly increased, and the Bax/Bcl-2 ratio was decreased in RIPC-EXOc group ( P<0.05). Compared with RIPC+ EXOc group, the cell viability was significantly decreased, and the Bax/Bcl-2 ratio was increased in RIPC+ P-EXOc group ( P<0.05). Conclusions:Propofol may abolish the myocardial protective effect of RIPC by decreasing the production and release of serum exosomes in rats.

Objective:To evaluate the role of exosomes in reduction of myocardial ischemia-reperfusion (I/R) injury (MIRI) by remote preconditioning of trauma (RPCT) in rats.Methods:This experiment was performed in 2 parts. In vivo experiment Adult male Sprague-Dawley rats, aged 8 weeks, weighing 250-300 g, were used. Six rats were selected and randomly divided into 2 groups ( n=3 each): control group and RPCT group. Rats in control group underwent thoracotomy only, while rats in RPCT group were subjected to an additional 4 cm transverse skin incision along the abdominal midline after thoracotomy. Blood samples were collected, and serum exosomes were isolated from blood samples and labeled as control exosomes and RPCT exosomes. The expression of exosomal surface marker proteins CD9 and heat shock protein 70 (HSP70) was determined by Western blot, and the serum exosome concentration was measured. Another 30 rats were selected and randomly assigned to 5 groups ( n=6 each): sham operation group (Sham group), I/R group, I/R+ RPCT group, I/R+ control exosomes group (I/R+ EXO-CON group), and I/R+ RPCT exosomes group (I/R+ EXO-RPCT group). The MIRI model was established by ligating the left anterior descending coronary artery for 30 min followed by 120 min of reperfusion in anesthetized animals. In I/R+ RPCT group, the MIRI model was prepared at 15 min after the end of RPCT. In I/R+ EXO-CON and I/R+ EXO-RPCT groups, control exosomes and RPCT exosomes 100 μg were administered via the jugular vein at 15 min before ischemia respectively. At the end of reperfusion, the myocardial infarct size was measured, serum concentrations of cardiac troponin T (cTnT) and lactate dehydrogenase (LDH) were determined, and the contents of malondialdehyde (MDA) and superoxide dismutase (SOD) in myocardial tissues were measured. The expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase-3 and cleaved caspase-3 was detected. In vitro experiment H9c2 cells were cultured in vitro and randomly divided into 4 groups ( n=6 each): control group (Con group), hypoxia/reoxygenation (H/R) group, H/R+ control exosomes group (H/R+ EXO-CON group), and H/R+ RPCT exosomes group (H/R+ EXO-RPCT group). The rats were subjected to 4 h of hypoxia followed by 16 h of reoxygenation to establish the H/R injury model. In H/R+ EXO-CON and H/R+ EXO-RPCT groups, control exosomes and RPCT exosomes 4 μg were added at 15 min before hypoxia respectively. The cell survival rate and concentration of lactate dehydrogenase (LDH) in the supernatant were measured, and the expression of Bax, Bcl-2, cleaved caspase-3 and caspase-3 was detected. Results:In vivo experiment Compared with control group, the expression of serum CD9 and HSP70 was significantly up-regulated, and the exosome concentration was increased in RPCT group ( P<0.05). Compared with Sham group, the serum concentrations of cTnT and LDH, percentage of myocardial infarct size, content of MDA in myocardial tissues, Bax/Bcl-2 ratio, and cleaved caspase-3/caspase-3 ratio were significantly increased, and the activity of SOD was decreased in I/R group ( P<0.05). Compared with I/R group, the serum cTnT and LDH concentrations, percentage of myocardial infarct size, content of MDA in myocardial tissues, Bax/Bcl-2 ratio and cleaved caspase-3/caspase-3 ratio were significantly decreased, and the activity of SOD was increased in I/R+ RPCT group ( P<0.05), and no significant changes were observed in the aforementioned parameters in I/R+ EXO-CON group ( P>0.05). There were no significant differences in the aforementioned parameters between I/R+ RPCT group and I/R+ EXO-RPCT group ( P>0.05). In vitro experiment Compared with Con group, the cell survival rate was significantly decreased, and the LDH concentration in the supernatant and Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were increased in H/R group ( P<0.05). Compared with H/R group, the cell survival rate was significantly increased, the LDH concentration in the supernatant, and Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were decreased in H/R+ EXO-CON group ( P<0.05), and no significant changes were found in the aforementioned parameters in H/R+ EXO-RPCT group ( P>0.05). Conclusions:The mechanism by which RPCT reduces MIRI may be related to the increased release of serum exosomes in rats.

Bacillus is a class of spore-producing Gram-positive bacteria that produce a variety of antimicrobial substances with different structures and functions. The application of the antimicrobial substances produced by Bacillus can effectively inhibit the activity of harmful bacteria and fungi and promote the sustainable development of green agriculture. The antimicrobial substances produced by Bacillus mainly include proteins, lipopeptides, polyketones, and polypeptides. This paper reviews the synthesis gene clusters, synthesis pathways, structures, and mechanisms of various antimicrobial substances produced by Bacillus and discusses the challenges in the industrial application of these antimicrobial substances. Furthermore, this paper clarifies the future research and development focuses and prospects the application prospects, and provides comprehensive theoretical support for the in-depth research and wide application of the antimicrobial substances produced by Bacillus.
Bacillus/genetics* ; Anti-Infective Agents/metabolism* ; Bacterial Proteins/genetics* ; Antimicrobial Peptides/biosynthesis* ; Lipopeptides/biosynthesis*

Objective:To evaluate the role of exosomes in propofol-induced elimination of cardioprotective effect of remote ischemic preconditioning (RIPC) in rats.Methods:This experiment was performed in 2 parts. In vivo experiment Forty-eight healthy SPF male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 5 groups using the random number table method: sham operation group (Sham group, n=12), ischemia-reperfusion (I/R) group ( n=12), RIPC group ( n=8), RIPC+ propofol group (RIPC+ P group, n=8), and propofol+ I/R group (P+ I/R group, n=8). The model of myocardial I/R injury was developed by ligating the left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion in anesthetized animals. Four cycles of 5-min ischemia induced by occlusion of the bilateral hind limbs with a tourniquet/5-min reperfusion served as the RIPC stimulus. Propofol was intravenously infused at a rate of 12 mg·kg -1·h -1 in RIPC+ P group (during RIPC) and in P+ I/R group (for 40 min). Exosomes from RIPC-treated and RIPC+ propofol-treated rats were extracted (RIPC-EXO and RIPC+ P-EXO respectively) for determination of the expression of surface markers of exosomes CD9 and HSP70. Another 24 rats were randomly selected, and the aforementioned exosomes were injected at 15 min before myocardial ischemia, resulting in RIPC-EXO+ I/R group ( n=12) and RIPC+ P-EXO+ I/R group ( n=12). At the end of reperfusion, the area of myocardial infarction was determined, the concentration of serum cardiac troponin I (cTnI) was measured by the enzyme-linked immunosorbent assay, and the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in myocardial tissues was detected by Western blot. Cell experiment H9c2 cells were cultured in vitro and divided into 4 groups ( n=6 each) using a random number table method: control group (C group), hypoxia-reoxygenation group (H/R group), RIPC-EXOc group and RIPC+ P-EXOc group. The cells were exposed to hypoxia for 4 h followed by reoxygenation for 16 h in H/R group. RIPC-EXO and RIPC+ P-EXO were added at a final concentration of 300 μg/ml before hypoxia in RIPC-EXOc group and RIPC+ P-EXOc group, respectively. The cell viability was determined using a cell counting kit-8 assay and the expression of Bax and Bcl-2 expression was detected by Western blot. Results:In vivo experiment Compared with RIPC-EXO group, the expression of CD9 and HSP70 was significantly down-regulated in RIPC+ P-EXO group ( P<0.05). Compared with Sham group, the percentage of the area of myocardial infarction was significantly increased, and the serum cTnI concentration and Bax/Bcl-2 ratio in myocardial tissues were increased in I/R group ( P<0.05). Compared with I/R group, the percentage of the area of myocardial infarction was significantly decreased in RIPC group and RIPC-EXO+ I/R group, the serum cTnI concentration and Bax/Bcl-2 ratio in myocardial tissues were significantly decreased in RIPC-EXO+ I/R group ( P<0.05), and no significant change was found in the percentage of the area of myocardial infarction in RIPC+ P group ( P>0.05). The percentage of the area of myocardial infarction was significantly larger in RIPC+ P group than in RIPC group ( P<0.05). Compared with RIPC-EXO+ I/R group, the percentage of the area of myocardial infarction was significantly increased, and the serum cTnI concentration and Bax/Bcl-2 ratio were increased in RIPC+ P-EXO+ I/R group ( P<0.05). Cell experiment Compared with C group, the cell viability was significantly decreased, and the Bax/Bcl-2 ratio was increased in H/R group ( P<0.05). Compared with H/R group, the cell viability was significantly increased, and the Bax/Bcl-2 ratio was decreased in RIPC-EXOc group ( P<0.05). Compared with RIPC+ EXOc group, the cell viability was significantly decreased, and the Bax/Bcl-2 ratio was increased in RIPC+ P-EXOc group ( P<0.05). Conclusions:Propofol may abolish the myocardial protective effect of RIPC by decreasing the production and release of serum exosomes in rats.

Objective:To evaluate the role of exosomes in reduction of myocardial ischemia-reperfusion (I/R) injury (MIRI) by remote preconditioning of trauma (RPCT) in rats.Methods:This experiment was performed in 2 parts. In vivo experiment Adult male Sprague-Dawley rats, aged 8 weeks, weighing 250-300 g, were used. Six rats were selected and randomly divided into 2 groups ( n=3 each): control group and RPCT group. Rats in control group underwent thoracotomy only, while rats in RPCT group were subjected to an additional 4 cm transverse skin incision along the abdominal midline after thoracotomy. Blood samples were collected, and serum exosomes were isolated from blood samples and labeled as control exosomes and RPCT exosomes. The expression of exosomal surface marker proteins CD9 and heat shock protein 70 (HSP70) was determined by Western blot, and the serum exosome concentration was measured. Another 30 rats were selected and randomly assigned to 5 groups ( n=6 each): sham operation group (Sham group), I/R group, I/R+ RPCT group, I/R+ control exosomes group (I/R+ EXO-CON group), and I/R+ RPCT exosomes group (I/R+ EXO-RPCT group). The MIRI model was established by ligating the left anterior descending coronary artery for 30 min followed by 120 min of reperfusion in anesthetized animals. In I/R+ RPCT group, the MIRI model was prepared at 15 min after the end of RPCT. In I/R+ EXO-CON and I/R+ EXO-RPCT groups, control exosomes and RPCT exosomes 100 μg were administered via the jugular vein at 15 min before ischemia respectively. At the end of reperfusion, the myocardial infarct size was measured, serum concentrations of cardiac troponin T (cTnT) and lactate dehydrogenase (LDH) were determined, and the contents of malondialdehyde (MDA) and superoxide dismutase (SOD) in myocardial tissues were measured. The expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase-3 and cleaved caspase-3 was detected. In vitro experiment H9c2 cells were cultured in vitro and randomly divided into 4 groups ( n=6 each): control group (Con group), hypoxia/reoxygenation (H/R) group, H/R+ control exosomes group (H/R+ EXO-CON group), and H/R+ RPCT exosomes group (H/R+ EXO-RPCT group). The rats were subjected to 4 h of hypoxia followed by 16 h of reoxygenation to establish the H/R injury model. In H/R+ EXO-CON and H/R+ EXO-RPCT groups, control exosomes and RPCT exosomes 4 μg were added at 15 min before hypoxia respectively. The cell survival rate and concentration of lactate dehydrogenase (LDH) in the supernatant were measured, and the expression of Bax, Bcl-2, cleaved caspase-3 and caspase-3 was detected. Results:In vivo experiment Compared with control group, the expression of serum CD9 and HSP70 was significantly up-regulated, and the exosome concentration was increased in RPCT group ( P<0.05). Compared with Sham group, the serum concentrations of cTnT and LDH, percentage of myocardial infarct size, content of MDA in myocardial tissues, Bax/Bcl-2 ratio, and cleaved caspase-3/caspase-3 ratio were significantly increased, and the activity of SOD was decreased in I/R group ( P<0.05). Compared with I/R group, the serum cTnT and LDH concentrations, percentage of myocardial infarct size, content of MDA in myocardial tissues, Bax/Bcl-2 ratio and cleaved caspase-3/caspase-3 ratio were significantly decreased, and the activity of SOD was increased in I/R+ RPCT group ( P<0.05), and no significant changes were observed in the aforementioned parameters in I/R+ EXO-CON group ( P>0.05). There were no significant differences in the aforementioned parameters between I/R+ RPCT group and I/R+ EXO-RPCT group ( P>0.05). In vitro experiment Compared with Con group, the cell survival rate was significantly decreased, and the LDH concentration in the supernatant and Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were increased in H/R group ( P<0.05). Compared with H/R group, the cell survival rate was significantly increased, the LDH concentration in the supernatant, and Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were decreased in H/R+ EXO-CON group ( P<0.05), and no significant changes were found in the aforementioned parameters in H/R+ EXO-RPCT group ( P>0.05). Conclusions:The mechanism by which RPCT reduces MIRI may be related to the increased release of serum exosomes in rats.

Objective:To evaluate the effect of obesity factor on myocardial ischemia-reperfusion injury and the role of transient receptor potential ankyrin 1 (TRPA1) expression in mice.Methods:Twenty-four SPF healthy male C57BL/6J mice, aged 6-7 weeks, weighing 18-21 g, were divided into 2 groups ( n=12 each) using a random number table method: common diet group (CD group) and high fat diet group (HFD group). CD group was fed a common diet and HFD group was fed a high-fat diet supplied with 60% fat for 12 weeks, and the body weight of mice was recorded every week. The mice in CD group were then divided into 2 groups ( n=6 each) using a random number table method: CD+ solvent group (CD+ C group) and CD+ TRPA1 agonist allyl isothiocyanate (AITC) group (CD+ A group). The mice in group HFD were divided into 2 groups ( n=6 each) using a random number table method: HFD+ solvent group (HFD+ C group) and HFD+ AITC group (HFD+ A group). At 13th week, AITC 25 mg·kg -1·d -1 was intragastrically administered based on the original diet in CD+ A group and HFD+ A group, and the equal volume of solvent was given in CD+ C and HFD+ C groups. At 14th week, myocardial I/R injury was established by occlusion of the left coronary artery for 30 min followed by 120-min reperfusion. The body weight, epididymal fat, mesenteric fat and perirenal fat were recorded, and the levels of LDH, triglyceride and cholesterol in serum were determined. The percentage of myocardial infarct size was calculated. The expression of TRPA1, Bax and Bcl-2 was detected by Western blot, and the ratio of Bax/Bcl-2 was calculated. Results:Compared with CD+ C group, the expression of TRPA1 in myocardial tissues was significantly up-regulated ( P<0.05), and no significant differences were found in the other parameters in CD+ A group ( P>0.05), and the concentrations of serum triglyceride and cholesterol, body weight, weight of epididymal fat, mesenteric fat, perirenal fat and percentage of myocardial infarct size were significantly increased, the expression of TRPA1 in myocardial tissues was down-regulated, and the concentration of LDH and ratio of Bax/Bcl-2 were increased in HFD+ C group ( P<0.05). Compared with HFD+ C group, the concentrations of serum triglyceride and cholesterol, body weight, weight of epididymal fat, mesenteric fat, perirenal fat and percentage of myocardial infarct size were significantly decreased, the expression of TRPA1 in myocardial tissues was up-regulated, and the concentration of LDH and ratio of Bax/Bcl-2 were decreased in HFD+ A group ( P<0.05). Conclusions:Obesity can aggravate myocardial ischemia-reperfusion injury in mice, and the down-regulated myocardial TRPA1 expression is involved in this process.

OBJECTIVE: To explore the genetic basis of a patient with clinically suspected Loeys-Dietz syndrome (LDS). METHODS: A child who had presented at Beijing Anzhen Hospital in September 2018 was selected as the study subject. Clinical data and family history of the patient were collected, along with peripheral blood samples of the proband and his parents. Whole exome sequencing (WES) was carried out through next-generation sequencing. RESULTS: Candidate variants were searched through bioinformatic analysis focusing on genes associated with hereditary aortic aneurysms. Candidate variant was verified by Sanger sequencing. The patient was found to have cardiovascular abnormalities including early-onset aortic dilatation and coarctation, and LDS syndrome was suspected. WES revealed that he has harbored a heterozygous c.1526G>T missense variant of the TGFBR2 gene. The same variant was not found in either parent and was predicted as likely pathogenic (PM1+PM2_Supporting+ PM6+PP3+PP4) based on the guidelines from the American College for Medical Genetics and Genomics (ACMG). CONCLUSION The TGFBR2 c.1526G>T variant probably underlay the LDS in this patient and was unreported previously in China. Above finding has enriched the mutational spectrum of the TGFBR2 gene associated with the LDS and provided a basis for the genetic counseling for the patient.
Child ; Humans ; Male ; China ; Computational Biology ; Family ; Loeys-Dietz Syndrome/genetics* ; Mutation ; Receptor, Transforming Growth Factor-beta Type II/genetics*

From the perspectives of the pathogenesis, diagnosis and treatment of cardiac graft vasculopathy(CGV), this review summarized the current understanding and cognition of its pathology, monitoring, diagnosis and treatment to provide rationales for better survivals of CGV.

Patent foramen ovale (PFO) is the most prevalent congenital heart disease, often accompanied by neurological symptoms as migraine, unexplained dizziness, and even anxiety and depression. Recent research findings indicate that the pathogenesis of neurological complications related to PFO involves abnormal embolism hypothesis, vasoactive substance hypothesis, impaired cerebral blood flow regulation and genetic inheritance. Treatments include primarily encompass pharmacological intervention and foramen ovale occlusion. This article summarizes the aforementioned research progress in order to provide clinical guidance for managing nervous system complications related to PFO.