Integrating Network Pharmacology and Experimental Validation to Investigate the Mechanism of Qushi Huatan Decoction Against Coronary Heart Disease.

Purpose: This study was designed to evaluate the effect and mechanism of the Qushi Huatan (QSHT) decoction against coronary heart disease (CHD) through network pharmacology and experimental verification. Methods: In the present study, the active ingredients of the QSHT decoction were identified by ultra performance liquid chromatography/tandem mass spectrometry (UPLC/MS), then the potential ingredients and coronary heart disease targets were predicted using the SwissTarget Prediction database and the database of Genecards and OMIM database, respectively. A herb-compound-target network was constructed using Cytoscape. GO and KEGG enrichment analysis were performed using the ClusterProfiler data package of R software. Molecular docking was used to predict the core targets of QSHT against CHD. In addition, we used a myocardial infarction (MI) and high-fat diet ApoE-/- mice model to investigate the cardioprotective effects of QSHT. Western blotting and immunochemistry were used to verify the core targets and the signaling pathway. Results: A total of 68 active ingredients were found in the QSHT decoction. Network pharmacology indicated 28 targets and 147 signal pathways, including AKT1, HIF-1α, GSK-3ß, TLR4 and NF-κB, those key targets were also verified by molecular docking. The results of GO and KEGG enrichment analysis showed that the targets of QSHT against CHD were largely associated with inflammatory and oxidative stress, and AKT/HIF-1α and TLR4/NF-κB pathways might be key functional pathways. In vivo, QSHT significantly improved cardiac function and attenuated fibrosis and inflammation. Furthermore, QSHT could significantly inhibit the expression of HIF-1α, TLR4, phosphorylation of AKT1, GSK-3ß and NF-κB after MI in ApoE-/- mice. Conclusion: Based on network pharmacology, molecular docking and experimental verification, this study demonstrated that QSHT could improve cardiac function and attenuate cardiac fibrosis by regulating TLR4/NF-κB and AKT/HIF-1α signaling pathway in post- MI and high-fat diet ApoE-/- mice.

Circ_0060551 promotes the migration and invasion of cervical cancer by upregulating TPD52.

PROBLEM: Cervical cancer has been recognized as the second most common cancer in women worldwide. Previous studies have reported that some circular RNAs (circRNAs) can influence the progression of cervical cancer. However, more researches are still required to unveil the underlying mechanism of how circRNAs regulate the progression of such cancer. We aimed at unveiling the mechanism of how circ_0060551 effected the progression of cervical cancer. METHOD OF STUDY: RT-qPCR and western blot assays were used to detect the expression and protein levels. Mechanism experiments were conducted to investigate the relationship among circ_0060551, TPD52, miR-520a-5p and ELAVL1. Rescue assays were mainly carried out to verify how circ_0060551 influenced the migration and invasion of cervical cancer cells. RESULTS: According to the results, circ_0060551 was upregulated in cervical cancer cells and could promote the migration and invasion of cells via TPD52. In addition, circ_0060551 could up-regulate TPD52 expression through a ceRNA model to target miR-520a-5p. Moreover, circ_0060551 could stabilize the mRNA expression of TPD52 via recruiting ELAVL1. CONCLUSION: Our study proved that circ_0060551 could promote the migration and invasion of cervical cancer cells.

Identifying Circulating Tumor DNA Mutations Associated with Neoadjuvant Chemotherapy Efficacy in Local Advanced Breast Cancer.

Circulating tumor DNA (ctDNA) correlates with tumor burden and provides early detection of treatment response and tumor genetic alterations in breast cancer. Neoadjuvant chemotherapy (NACT) has become standard therapy for local advanced breast cancer (LABC). The aim of our study was to investigate plasma ctDNA as a prognostic marker for outcome in patients with LABC treated with NACT. A total of 56 patients with LABC were involved in this study. ctDNA mutations were investigated by using a 100 gene panel-target capture next-generation sequencing. The patients then received standard NACT therapy: adriamycin and cyclophosphamide and paclitaxel (AC-T) or AC-TH (AC-T+ Trastuzumab) regimen. The efficacy of NACT was evaluated by Miller-Payne grading system. A predictive and weight model was used to screen ctDNA point mutation biomarkers for NACT. The ctDNA mutational profile of LABC patients was identified. For nonsynonymous mutations, the top 5 mutated genes were MTHFR (51/56, 91.1%), XPC (50/56, 89.3%), ABCB1 (48/51, 94.1%), BRCA2 (38/56, 67.9%), and XRCC1 (38/56, 67.9%). In addition, the mutation frequencies of PIK3CA and TP53 were 32.1% (18/56) and 26.8% (15/56), respectively. The predictive model indicated that XRCC1 44055726 (TG>-) mutation (25/56, 44.6%) was significantly associated with Miller-Payne 4-5 and Miller-Payne 3-5 responses. While mTOR 11249132(G>C) mutation (23/56, 41.1%) was associated with Miller-Payne 1-4 or Miller-Payne 1-3 responses. Furthermore, XRCC1 44055726 (TG>-) accompanied by mTOR wild type predicted a good NACT efficacy in all response classification systems. The ROC curves to discriminate good neoadjuvant chemotherapy efficiency (Miller-Payne 4-5) and poor efficiency (Miller-Payne 1-3) were created, and AUC value was 0.77. Our results suggested that ctDNA mutation of XRCC1 44055726 (TG>-) might be a positive biomarker for NACT therapy in LABC, while mTOR 11249132(G>C) mutation was potentially associated with NACT resistance.

Effect of pharmaceutical care on the treatment of COVID-19: A protocol for systematic review and meta analysis.

BACKGROUND: We aimed to conduct a meta-analysis to assess the effect of pharmaceutical care on the treatment of coronavirus disease 2019 (COVID-19). METHODS: All case-controlled studies related to pharmaceutical care on the treatment of COVID-19 will be included in this review. We will use index words related to pharmaceutical care and COVID-19 to perform literature searches in PubMed, Embase, MEDLINE, CNKI, and Wanfang databases, to include articles indexed as of October 20, 2020 in English and Chinese language. Two reviewers will select trials independently for inclusion and assess trial quality. Two pairs of review authors will independently extract information for each included trials. Primary outcomes are clinical outcomes, average hospital stays, costs, patient satisfaction, and incidence of adverse drug reactions. We will evaluate the risk of bias of the included studies based on Cochrane assessment tool. Revman 5.3 (the Cochrane collaboration, Oxford, UK) will be used for heterogeneity assessment, generating funnel-plots, data synthesis, subgroup analysis, and sensitivity analysis. RESULTS: We will provide targeted and practical results assessing the effect of pharmaceutical care on the treatment of COVID-19. CONCLUSION: The stronger evidence about the effect of pharmaceutical care on the treatment of COVID-19 will be provided for clinicians. SYSTEMATIC REVIEW REGISTRATION NUMBER: PROSPERO CRD42020214223 ETHICS AND DISSEMINATION:: There is no need for ethical approval, and the review will be reported in a peer-reviewed journal.

Ultrasonographic Diagnosis of Lipomatosis of Nerve: A Review of Ultrasonographic Finding for 8 Cases.

OBJECTIVE: Lipomatosis of nerve (LN) is a rare tumor-like condition with epineural and perineural infiltration by adipose and fibrous tissue. The purpose was to analyze the ultrasonographic findings of LN involving upper limb peripheral nerves. METHODS: This was a retrospective analysis of a series of 8 patients with LN involving upper-limb peripheral nerves between 2013 and 2019. All patients underwent preoperative ultrasonography for the upper-extremity nerves and were diagnosed as LN by surgery. The clinical manifestations, ultrasonography characteristics, and accuracy were analyzed. RESULTS: In this series, LN was involved in 10 peripheral nerves from 8 patients. The median nerve was the most commonly affected nerve (60%). Four cases presented macrodactyly combined with masses from distal forearm and extending to wrist and palm areas. Among 8 patients, 5 cases were diagnosed with LN by preoperative ultrasonography, an accuracy of 62.5%. Axial ultrasonic imaging showed the punctate hypoechoic fascicles was embedded in hyperechoic adipose tissue in the "lotus root-like" appearance; longitudinal ultrasonic imaging showed the strip hypoechoic fascicles alternates with hyperechoic adipose tissue in the "cable-like" appearance. Meanwhile, ultrasonic imaging showed the thickened of adipose tissue around the affected nerve and the enlargement of flexor tendons in some patients. CONCLUSIONS: Ultrasonography has the potential to be a useful tool for the noninvasive examination of LN. The possibility of LN should be considered in patients with a mass in wrist and palm, macrodactyly, or syndactyly. Our finding may benefit the preoperative differential diagnosis with common nerve tumors.

The Effectiveness of Ultrasonography in the Diagnosis of Spontaneous Hourglasslike Constriction of Peripheral Nerve in the Upper Extremity.

OBJECTIVE: To investigate the ultrasonographic characteristics in hourglasslike constriction of peripheral nerve in the upper extremity and to evaluate the value of ultrasonography in the diagnosis. METHODS: Nineteen patients with hourglasslike constriction of peripheral nerve in the upper extremity underwent ultrasonography and the results were compared with surgery. The ultrasonographic characteristics, the accurate rate, and the relation between the ultrasonography and surgery were analyzed. RESULTS: There were 22 affected nerves involved in 19 patients, including 17 radial neuropathies, 4 median neuropathies, and 1 musculocutaneous neuropathy. The accuracy rate of ultrasonography in diagnosing hourglasslike constriction of upper limb nerve was 87.93%. Ultrasonography showed that the constriction sites were completely consistent with the operation. The ultrasonography characteristics of hourglasslike constriction of upper limb nerves were hourglasslike nerve incompleteness or complete constriction, and the nerves at both ends were thickened, and no compression structure was seen around. All lesions with complete constriction diagnosed by ultrasonography were treated with resection of the lesion with or without graft. In addition, 71.43% with incomplete constriction were treated with neurolysis, and 28.57% with resection of the lesion with direct repair. CONCLUSIONS: Ultrasonography could be used as a routine noninvasive examination for hourglasslike constriction of upper limb nerves. Ultrasonography suggests that resection of the lesion rather than neurolysis should be considered in the treatment of complete constriction. For patients with clinical symptoms, ultrasonography showed local nerve enlargement but no constriction; clinicians should be prompted to explore carefully during operation to avoid missing nerve hourglasslike constriction.

MicroRNA-21 increases the expression level of occludin through regulating ROCK1 in prevention of intestinal barrier dysfunction.

OBJECTIVE: The aim of this study is to investigate the role of molecular mechanism of microRNA (miR)-21 on tight junction (TJ)-proteins and its protective effects on the intestinal barrier. METHODS: TJ proteins and target genes expression were analyzed in miR-21 inhibition and overexpression NCM460 cell lines. To further verify the role of miR-21, the mmu-miR-21 intestinal epithelial conditional knockout (IKO) mice model was established. MiR-21 expression was detected in clinical specimens of acute stercoral obstruction patients. RESULTS: Rho-associated protein kinase 1 (ROCK1) were identified as target genes of miR-21. There is a negative correlation between miR-21 expression level and TJ proteins levels. TJ protein and ROCK1 were significantly decreased in miR-21 IKO mice, which presented intestinal inflammation response and intestinal barrier dysfunction (both P < 0.05). Determination of clinical samples showed consistent results with NCM460 cell line and miR-21 IKO mice. CONCLUSIONS: MiR-21 could be a protective factor of intestinal barrier dysfunction, which promoting the expression of TJ protein by targeting ROCK1 in vivo and in vitro.

Shensong Yangxin capsule reduces atrial fibrillation susceptibility by inhibiting atrial fibrosis in rats with post-myocardial infarction heart failure.

PURPOSE: Shensong Yangxin (SSYX) capsule is a traditional Chinese medicine that has been used widely to treat cardiac arrhythmia. This study aimed to assess whether SSYX prevents atrial fibrillation (AF) after chronic myocardial infarction (MI)-induced heart failure and to determine the underlying mechanisms. MATERIALS AND METHODS: The study included 45 male Sprague Dawley rats. The rats underwent MI induction or sham surgery. One week after MI induction surgery, we performed serial echocardiography and administered SSYX capsule to some rats that experienced MI. After 4 weeks of treatment, AF inducibility was assessed with transesophageal programmed electrical stimulation technology. Additionally, multielectrode array assessment, histological analysis, and Western blot analysis were performed. RESULTS: AF inducibility was significantly lower in SSYX rats than in MI rats (33.3% vs 73.3%, P<0.05). Additionally, conduction velocities in the left atrium were greater in SSYX rats than in MI rats. Moreover, SSYX decreased left atrial fibrosis, downregulated TGF-ß1, MMP-9, TIMP-I, and type I and III collagen expressions, and inhibited the differentiation of cardiac fibroblasts to myofibroblasts. CONCLUSION: SSYX reduces AF inducibility after MI by improving left atrial conduction function via the inhibition of left atrial fibrosis. It prevents the development of an MI-induced vulnerable substrate for AF.

Shengmai San-derived herbal prevents the development of a vulnerable substrate for atrial fibrillation in a rat model of ischemic heart failure.

OBJECTIVE: The study aimed to investigate whether a Shengmai San-derived herbal, Fumai granule (FM), which had a preventive effect on atrial fibrillation (AF) in myocardial infarction (MI)-induced heart failure (HF) rat and to determine the underlying mechanisms. DESIGN AND METHODS: MI was induced by a ligation of the left anterior descending coronary artery. One week after MI surgery, FM was gavaged for 4 weeks. AF inducibility was detected by transesophageal programmed electrical stimulation technology. Multielectrode array measurements, echocardiogram, histology, and western blotting were performed. RESULTS: The FM-treated group had lower rates of AF inducibility and shorter AF duration compared to the MI group. FM improved the conduction velocity and homogeneity, decreased left atrial positive fibrosis areas and expression of type I and III collagen, inhibited cardiac fibroblast to myofibroblast differential, and increased the expression of connexin 43 and connexin 40 in the left atrium. CONCLUSIONS: These results suggest that FM reduced the AF inducibility after MI by improving the left atrial conduction function via inhibiting left atrial fibrosis and increasing the expression of connexin, indicating its benefit in preventing the MI-induced vulnerable substrate for AF.

Matrine reduces susceptibility to postinfarct atrial fibrillation in rats due to antifibrotic properties.

This study aimed to investigate whether matrine could prevent atrial fibrillation (AF) after myocardial infarction by reducing left atrial fibrosis, and to determine the underlying mechanisms in isolated cardiac fibroblasts (CFs). Five weeks after MI, matrine-treated rats had lower rates of AF inducibility and shorter AF duration than MI rats. Matrine improved the left atrial conduction velocity and homogeneity. Matrine decreased the fibrosis positive areas and the protein levels of type I collagen and type III collagen in the left atrium. Matrine inhibited CFs differentiation to myofibroblasts and the expression of transforming growth factor-beta 1 and matrix metalloproteinase 9. In vitro, matrine inhibited the CFs proliferation, migration, differentiation, and secretion ability. These in vitro and in vivo data demonstrated that matrine has the potential to reduce susceptibility to AF after MI due, at least in part, to reduced atrial fibrosis via inhibiting CFs proliferation, migration, differentiation, and secretion ability.

Attenuation of ischemia/reperfusion-induced inhibition of the rapid component of delayed rectifier potassium current by Isosteviol through scavenging reactive oxygen species.

Isosteviol has been demonstrated to play a protective role during ischemia reperfusion (I/R) myocardial infarction. However, the underlying electrophysiological mechanisms of isosteviol are still unknown. Our previous study showed that the rapid component of the delayed rectifier potassium channel (IKr) plays an important role in the prolongation of I/R-induced QT interval-related arrhythmia. This study aimed to investigate whether isosteviol could attenuate I/R-induced prolongation of the action potential duration (APD) along with inhibition of IKr, and we aimed to clarify the electrophysiological mechanism of isosteviol to determine its cardioprotective effects in guinea pigs. We observed that the APD90 were 298.5±41.6ms in control, 528.6±56.7ms during I/R, and reduced to 327.8±40.5ms after 10µmol/L of isosteviol treatment. The IKr currents were 1.44±0.06 pA·pF-1in the control group, 0.50±0.07pA·pF-1during I/R, and recovered to 1.20±0.12pA·pF-1after 10µmol/L of isoteviol treatment. Moreover, isosteviol reduced the over-production of reactive oxygen species (ROS) during I/R. Importantly, isosteviol does not affect the IKr and human ether-a-go-go-related gene currents of normal cardiomyocytes. It attenuated the I/R-induced inhibition of IKr due to reduced over-production of ROS. Furthermore, isosteviol is safe and has no cardiotoxicity, and it might be beneficial for coronary reperfusion therapy.

Inhibition of rapid delayed rectifier potassium current (IKr) by ischemia/reperfusion and its recovery by vitamin E in ventricular myocytes.

Ischemia/reperfusion (I/R) induces prolongation of QT interval and action potential duration (APD), which is a major cardiac electrical disorder in patients with arrhythmias. However, the mechanism of QT interval prolongation induced by I/R remains unclear. In the present study, we hypothesized that the rapid component of delayed rectifier potassium (IKr) channel plays an important role in I/R-induced QT interval prolongation. We observed a marked attenuation of IKr and a significant prolongation of action potential duration (APD) in a simulated I/R system with sodium dithionite (Na2S2O4) in ventricular myocytes of guinea pigs. The IKr current density was inhibited by 64% and APD increased by 87% respectively. Moreover, the inhibition of IKr is primarily ascribed to overproduction of reactive oxygen species (ROS) by I/R, which can be partly reversed by antioxidant vitamin E (100µmol/L). The value of IKr tail current density increased from 0.516±0.040 pA/pF in I/R to 0.939±0.091 pA/pF when treated with vitamin E. Moreover, we also demonstrated that QTc interval was increased by I/R and reversed by Vitamin E in isolated guinea pig hearts. In conclusion, the inhibition of IKr is one of the underlying mechanisms of prolongation of QT interval and APD in I/R. Vitamin E might have a benefit in coronary reperfusion therapy.

Isosteviol Sensitizes sarcKATP Channels towards Pinacidil and Potentiates Mitochondrial Uncoupling of Diazoxide in Guinea Pig Ventricular Myocytes.

KATP channel is an important mediator or factor in physiological and pathological metabolic pathway. Activation of KATP channel has been identified to be a critical step in the cardioprotective mechanism against IR injury. On the other hand, desensitization of the channel to its opener or the metabolic ligand ATP in pathological conditions, like cardiac hypertrophy, would decrease the adaption of myocardium to metabolic stress and is a disadvantage for drug therapy. Isosteviol, obtained by acid hydrolysis of stevioside, has been demonstrated to play a cardioprotective role against diseases of cardiovascular system, like anti-IR injury, antihypertension, antihyperglycemia, and so forth. The present study investigated the effect of isosteviol (STV) on sarcKATP channel current induced by pinacidil and mitochondrial flavoprotein oxidation induced by diazoxide. Our results showed that preincubating cells with STV not only increased the current amplitude and activating rate of sarcKATP channels induced by pinacidil but also potentiated diazoxide-elicited oxidation of flavoprotein in mitochondria.

Intercellular adhesion molecule-1 expression in activated eosinophils is associated with mucosal remodeling in nasal polyps.

Nasal polyposis (NP) is characterized by chronic mucosal inflammation with infiltrating eosinophils. Eosinophil­mediated tissue remodeling may be involved in NP pathogenesis; therefore, improved understanding of tissue remodeling may result the identification of novel pathways and therapeutic strategies. The present study aimed to investigate the pathological changes occurring during tissue remodeling in NP, in order to assess the role of intercellular adhesion molecule­1 (ICAM­1) in localized tissue remodeling and the potential association between ICAM­1 expression and markers of eosinophil activation. A total of 28 eligible patients and 10 healthy controls participated in the current study. Nasal mucosal tissues of these subjects were retrospectively evaluated for mucosal remodeling using histopathological staining. ICAM­1 and eosinophil cationic protein (ECP) expression levels were determined by immunohistochemical analysis. Compared with the healthy controls, all the specimens from NP patients presented substantial epithelial damage, skewed cellular distribution with a reduced density of goblet cells, an increased density of subepithelial gland and increased subepithelial collagen deposition. In addition, the NP specimens exhibited significantly higher eosinophil infiltration and ICAM­1 expression compared with the controls. Positive correlations were observed between ICAM­1 and ECP expression levels (P=0.010), as well as between extracellular collagen deposition and ICAM­1 (P=0.010) and ECP (P=0.012) expression levels in the NP specimens, but not in the control specimens. Morphological evidence demonstrated eosinophil­mediated tissue remodeling in NP tissues. ICAM­1 expression in activated eosinophils was associated with NP remodeling, indicating the possibility that ICAM­1 may regulate NP remodeling.

Clinical significance of the stem cell gene Oct-4 in cervical cancer.

This study aims to investigate the association between the expression of Oct-4 and the biological behavior or prognosis of cervical cancer. Serum-free suspension culture technology was used to select a suspension of microspheres that can stabilize clones. The tumorigenicity of the microsphere suspension was analyzed in NOD/SCID mice. Microarray analysis was used to detect the specific expression of genes in the microsphere suspension. The expression of Oct-4 was detected by immunohistochemistry, and the correlation between Oct-4 expression and clinical pathological prognostic indicators was analyzed in cervical cancer. The expression of the following genes was significantly different between the experimental and control groups: stem cell differentiation (CD44 and Oct-4), markers cell cycle regulators (APC), cell cycle regulators (MYC), and self-renewal markers (MYST2, NEUROG2, and SOX1). The expression of Oct-4 was significantly higher in cervical cancer tissues than in adjacent normal tissues and was significantly related to differentiation, clinical stage, and lymph node metastasis. The 5-year survival rate of patients with Oct-4-positive expression was lower than that of patients with Oct-4-negative expression (36.7 vs. 67.7 %, respectively; P=0.001). Cox regression analysis revealed that clinical stage, lymph node metastasis, and Oct-4 were independent prognostic factors in cervical cancer (P=0.031, 0.012, and 0.001, respectively). Our results showed that Oct-4 was highly expressed in cervical cancer stem cells; Oct-4 expression was associated with biological behavior and was an independent prognostic factor in cervical cancer. Therefore, it may represent a potential target for cervical cancer treatment.

[Comparison of the effect of fluoxetine combined with hormone replacement therapy (HRT) and single HRT in treating menopausal depression].

OBJECTIVE: To compare the treatment effects between fluoxetine plus hormone replacement therapy (HRT) and HRT alone on menopausal depression. METHODS: A total of 54 women with climacteric symptoms and depression were enrolled and randomly divided into two groups. The fluoxetine plus HRT group received fluoxetine (20 mg, qd, po) and cyclic use of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg, while HRT group was assigned to receive cyclic use of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg only. All subjects were interviewed and evaluated with Hamilton Depression score (HAMD) and Kupperman menopause index (KMI) at week 0, 1, 2, 3, 4, 6 and 8 of the treatment. RESULTS: There was no difference between two groups in HAMD and KMI before the trial (P > 0.05). HAMD scores at the 8th week in fluoxetine plus HRT group and HRT group were 3.0 +/- 2.3 and 11.2 +/- 5.8 respectively, being significantly different (P < 0.001). At the 6th week of treatment KMIs in fluoxetine plus HRT group and HRT group were 9 +/- 6 and 14 +/- 7 respectively, also significantly different (P < 0.05). The healing rates of fluoxetine plus HRT group and HRT group were 92% and 48% respectively, which were statistically different by Chi square test (P < 0.001). CONCLUSIONS: The effect of fluoxetine plus HRT on menopausal depression is significantly superior than that of HRT only and the difference becomes more obvious with treatment time. Both groups could ameliorate the climacteric symptoms with the effect of fluoxetine plus HRT more obvious.