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Objective: To reveal the similarities and differences in myocardial metabolic characteristics between heart failure with preserved ejection fraction (HFpEF) and heart failure with reduced ejection fraction (HFrEF) mice using metabolomics. Methods: The experimental mice were divided into 4 groups, including control, HFpEF, sham and HFrEF groups (10 mice in each group). High fat diet and Nω-nitroarginine methyl ester hydrochloride (L-NAME) were applied to construct a"two-hit"HFpEF mouse model. Transverse aortic constriction (TAC) surgery was used to construct the HFrEF mouse model. The differential expression of metabolites in the myocardium of HFpEF and HFrEF mice was detected by untargeted metabolomics (UHPLC-QE-MS). Variable importance in projection>1 and P<0.05 were used as criteria to screen and classify the differentially expressed metabolites between the mice models. KEGG functional enrichment and pathway impact analysis demonstrated significantly altered metabolic pathways in both HFpEF and HFrEF mice. Results: One hundred and nine differentially expressed metabolites were detected in HFpEF mice, and 270 differentially expressed metabolites were detected in HFrEF mice. Compared with the control group, the most significantly changed metabolite in HFpEF mice was glycerophospholipids, while HFrEF mice presented with the largest proportion of carboxylic acids and their derivatives. KEGG enrichment and pathway impact analysis showed that the differentially expressed metabolites in HFpEF mice were mainly enriched in pathways such as biosynthesis of unsaturated fatty acids, ether lipid metabolism, amino sugar and nucleotide sugar metabolism, glycerophospholipid metabolism, arachidonic acid metabolism and arginine and proline metabolism. The differentially expressed metabolites in HFrEF mice were mainly enriched in arginine and proline metabolism, glycine, serine and threonine metabolism, pantothenate and CoA biosynthesis, glycerophospholipid metabolism, nicotinate and nicotinamide metabolism and arachidonic acid metabolism, etc. Conclusions: HFpEF mice have a significantly different myocardial metabolite expression profile compared with HFrEF mice. In addition, biosynthesis of unsaturated fatty acids, arachidonic acid metabolism, glycerophospholipid metabolism and arginine and proline metabolism are significantly altered in both HFpEF and HFrEF mice, suggesting that these metabolic pathways may play an important role in disease progression in both types of heart failure.
Subject(s)
Mice , Animals , Heart Failure/metabolism , Stroke Volume , Chromatography, Liquid , Tandem Mass Spectrometry , Metabolomics , Arachidonic Acids , ProlineABSTRACT
Objective: To investigate the impact of surgical treatment on quality of life in patients with locally recurrent rectal cancer (LRRC). Methods: A descriptive case series study was performed. The complete clinical data of 62 patients who met the diagnostic criteria of LRRC and treated by surgical procedures in Huashan Hospital of Fudan University from January 2012 to November 2019 were analyzed retrospectively. All the patients were followed up at least 12 months. Assessments of urinary function, sexual function, mobility function of lower limb and quality of life were documented. Patients with distant metastasis and surgical history of the urinary system were excluded. According to the criteria of Memorial Sloan Kettering Cancer Center (MSKCC), recurrence were divided into central (n=27), anterior (n=20), posterior (n=7), and lateral (n=8) subtypes. Baseline characteristics, surgical procedures and short-term complications were analyzed. International prostate symptom score (IPSS) and grade of voiding dysfunction were used to evaluate the urinary function. Higher score of IPSS and higher grade of voiding dysfunction indicated worse voiding function. Sexual function for both genders was assessed preoperatively and postoperatively. International index of erectile function-5 (IIEF-5) was used for assessment of male patients and higher score indicated better function. Female sexual function index (FSFI) was used in females and higher score indicated better function. Short-form health survey with 36 items (SF-36), yielding an 8-scale profile of functional health (physical functioning, role-physical, bodily pain, general health, vitality, social functioning, emotional health and mental health) was used to evaluate the quality of life. The higher score indicated the better quality of life. Results: All the operations of 62 patients completed successfully and R0 resection rate was 88.7% (55/62). Postoperative surgical complications occurred in 16 cases (25.8%), including 3 patients of Clavien-Dindo classification III. At postoperative 3-month, 42 patients without ileum cystectomy or ureterostomy suffered from different grade of voiding dysfunction. IPSS increased significantly after the surgery (before surgery: 12.36±4.75, after surgery: 18.40±4.77, t=-9.128, P<0.001). There was no significant difference among the subtypes (P>0.05). At postoperative 12-month, IIEF-5 decreased from 14 (0~25) to 9 (0~19) in males (Z=-5.174, P<0.001) and FSFI deceased from 8.4 (2.0-27.0) to 2.0 (2.0-18.4) in females (Z=-3.522, P<0.001). Scores of physical functioning and role-physical decreased significantly [physical functioning: before surgery 70 (35-85), after surgery 65 (30-80), Z=-3.685, P<0.001; role-physical: before surgery 50 (0-50), after surgery 25(0-75), Z=-4.065, P<0.001], while those of social functioning role-emotional and mental health increased significantly after the surgery [social functioning: before surgery 44 (22-78), after surgery 56 (0-89), Z=-3.509, P<0.001; role-emotional: before surgery 17 (0-100), after surgery 33 (0-100), Z=-2.439, P=0.015; mental health: before surgery 40 (36-76), after surgery 52 (24-80), Z=-3.395, P<0.001]. All surgical procedures decreased the voiding function of LRRC patients and the sexual function of male patients (all P<0.01). However, only total pelvic exenteration and posterior pelvic exenteration decreased FSFI in female patients [before surgery: 8.4 (2.0-27.0) after surgery: 2.0 (2.0-18.4), Z=-2.810, P=0.005]. Conclusions: Multi-visceral resection in LRRC patients may damage voiding and sexual function. However, successful and effective surgical treatment can improve the psychosocial health of LRRC patients.
Subject(s)
Female , Humans , Male , Neoplasm Recurrence, Local , Quality of Life , Rectal Neoplasms/surgery , Rectum , Retrospective StudiesABSTRACT
ObjectiveExosomes secreted by BMSC overexpressing GATA-4 gene (BMSCGATA-4-exosome) can promote the differentiation of BMSC into cardiomyocyte-like cells, thereby improve cardiac function after myocardial infarction. However, the molecular mechanism of BMSCGATA-4-exosome in cardiomyocyte-like cell differentiation is unknown. The effect of the secretion of BMSCGATA-4 exosome from bone marrow mesenchymal stem cells (BMSC) in the differentiation of stem cells into cardiomyocytes was determined in miRNA-673-5p/Tsc-1 axis dependent manner.MethodsMouse models of myocardial infarction were established and divided into seven groups. Simulation group (BMSCmiR-673-5p-mimic exosome), inhibition group (BMSCmiR-673-5p-inhibitor exosome), GATA-4 group (BMSCGATA-4 exosome), empty vector group (BMSCempty vector exosome）, and BMSC group (BMSC exosome) were injected into the tail vein for 48 h, and the untreated and normal mice were used as the control group. Cardiac ultrasound was used to detect cardiac function in each group. miRNA-673-5p expression in myocardial infarction was detected using real-time polymerase chain reaction (RT-PCR). The myocardial tissues were extracted from the same myocardial infarction site. Myocardial-specific molecules, such as α-actin, Desmin, cTnT, and Cx43, were detected using RT-PCR. Western blot was used to determine the expression of the corresponding target gene of miRNA-673-5p, Tsc-1, Erk1/2, and Mef2c proteins.ResultsThe simulation group wan shown the most significantly improved myocardial function (P<0.05) with an expression peak of miRNA-673-5p in cardiomyocytes (P<0.05). The highest content of myocardial-specific molecules including α-actin, Desmin, cTnT, and Cx43 was found in the simulation group. The simulation group had the lowest expression of Tsc-1 in cardiomyocytes (P<0.05).ConclusionOverexpressed BMSCGATA-4 exosomes inhibit Tsc-1 expression through miRNA-673-5p to improve cardiac function during myocardial infarction.
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Objective: To predict B cell and T cell epitopes of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins. Methods: The sequences of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins which were derived from Orientia tsutsugamushi were analyzed by SOPMA, DNAstar, Bcepred, ABCpred, NetMHC, NetMHC II and IEDB. The 58-kDa tertiary structure model was built by MODELLER9.17. Results: The 22-kDa B-cell epitopes were located at positions 194-200, 20-26 and 143-154, whereas the T-cell epitopes were located at positions 154-174, 95-107, 17-25 and 57-65. The 47-kDa protein B-cell epitopes were at positions 413-434, 150-161 and 283-322, whereas the T-cell epitopes were located at positions 129-147, 259-267, 412-420 and 80-88. The 56-kDa protein B-cell epitopes were at positions 167-173, 410-419 and 101-108, whereas the T-cell epitopes were located at positions 88-104, 429-439, 232-240 and 194-202. The 58-kDa protein B-cell epitopes were at positions 312-317, 540-548 and 35-55, whereas the T-cell epitopes were located at positions 415-434, 66-84 and 214-230. Conclusions: We identified candidate epitopes of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins from Orientia tsutsugamushi. In the case of 58-kDa, the dominant antigen is displayed on tertiary structure by homology modeling. Our findings will help target additional recombinant antigens with strong specificity, high sensitivity, and stable expression and will aid in their isolation and purification.
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Objective: To predict B cell and T cell epitopes of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins. Methods: The sequences of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins which were derived from Orientia tsutsugamushi were analyzed by SOPMA, DNAstar, Bcepred, ABCpred, NetMHC, NetMHCⅡ and IEDB. The 58-kDa tertiary structure model was built by MODELLER9.17. Results: The 22-kDa B-cell epitopes were located at positions 194-200, 20-26 and 143-154, whereas the T-cell epitopes were located at positions 154-174, 95-107, 17-25 and 57-65. The 47-kDa protein B-cell epitopes were at positions 413-434, 150-161 and 283-322, whereas the T-cell epitopes were located at positions 129-147, 259-267, 412-420 and 80-88. The 56-kDa protein B-cell epitopes were at positions 167-173, 410-419 and 101-108, whereas the T-cell epitopes were located at positions 88-104, 429-439, 232-240 and 194-202. The 58-kDa protein B-cell epitopes were at positions 312-317, 540-548 and 35-55, whereas the T-cell epitopes were located at positions 415-434, 66-84 and 214-230. Conclusions: We identified candidate epitopes of 22-kDa, 47-kDa, 56-kDa and 58-kDa proteins from Orientia tsutsugamushi. In the case of 58-kDa, the dominant antigen is displayed on tertiary structure by homology modeling. Our findings will help target additional recombinant antigens with strong specificity, high sensitivity, and stable expression and will aid in their isolation and purification.
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Objective Exosomes secreted from mouse bone marrow mesenchymal stem cells (BMSC) overexpressing the cardiomyocyte transcription factor GATA-4 (BMSCGATA-4-exosome) may play a key role in repairing myocardial injury. This study aimed to investigate the molecular regulatory network of BMSCGATA-4-exosome for inhibiting the apoptosis of cardiomyocytes. Methods Exosomes extracted from GATA-4-overexpressing BMSCs of the mouse cultured with miR-330-3p-mimic were cultured with myocardial cells under hypoxic and serum-free conditions for 24 hours (the experimental group), the overexpressed GATA-4, empty vector and BMSCs were taken as the confounding factor control (CFC), the myocardial cells cultured under hypoxic and serum-free conditions for 24 hours were used as the positive control, and those cultured under the normal condition for 24 hours as the negative control. The apoptosis rates of myocardial cells in different groups were measured by flow cytometry, the expression levels of miR-330-3p in the myocardial cells determined by RT-PCR, and those of the corresponding miR-330-3p target gene Ap2m1 and transcriptional protein Cnot4 detected by Western blot. Results CD29 was expressed in 99.71% of the mouse BMSCs, CD44 in 97.28%, SCA-1 in 99.40%, and CD11b overexpressed in only 0.1%. The early apoptosis rate of myocardial cells was significantly higher in the experimental than in the negative control group ([7.90 ± 0.34]% vs [2.30 ± 0.09]%, P < 0.05) but lower than in the positive control ([51.48 ± 0.40]%), BMSC ([18.32 ± 3.03]%), empty vector ([16.99 ± 2.93]%) and overexpressed GATA-4 groups ([10.22 ± 0.35]%) (P < 0.05). The expression of miR-330-3p in the myocardial cells was markedly higher in the experimental ([396.10 ± 1.02]%) than in the negative control ([1.37 ± 0.33]%), positive control ([0.26±0.32]%), BMSC ([1.40 ± 0.42]%), empty vector ([1.41 ± 0.27]%) and overexpressed GATA-4 groups ([3.80 ± 0.62]%) (P < 0.05). The expressions of Ap2m1 and Cont4 in the myocardial cells were remarkably decreased in the experimental group compared with those in the other five groups (P < 0.05). Conclusion Overexpressed BMSCGATA-4-exosomes suppress the apoptosis of myocardial cells by inhibiting the expression of the Ap2m1 protein via miR-330-3p.
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Repeated IVF failure has been a pain pointin the assisted reproductive process.The fallopian tubehas long been considered as a conduit for transporta-tion,ignoring its important role in embryo implantation.The main reasons for the repeated failure of IVF areembryonic factors,endometrial receptivity and otherfactors.The article mainly focuses on the effects ofvarious types of fallopian tube lesions on the abovethree factors,and analyzes the relationship betweenvarious oviduct chronic inflammatory lesions and IVFfailure.IVFfallopian tube obstructionendome-