Progress of Exosomal MicroRNAs and Traditional Chinese Medicine Monomers in Neurodegenerative Diseases.

Exosomes, extracellular vesicles secreted by various cells, actively participate in intercellular communication by facilitating the exchange of crucial molecular information such as DNA, RNA, and lipids. Within this intricate network, microRNAs, endogenous non-coding small RNAs, emerge as pivotal regulators of post-transcriptional gene expression, significantly influencing the development of neurodegenerative diseases. The historical prominence of traditional Chinese medicine (TCM) in clinical practice in China underscores its enduring significance. Notably, TCM monomers, serving as active constituents within herbal medicine, assume a critical role in the treatment of neurodegenerative diseases, particularly in mitigating oxidative stress, inhibiting apoptosis, and reducing inflammation. This comprehensive review aims to delineate the specific involvement of exosomal microRNAs in various neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Huntington's disease, stroke, and amyotrophic lateral sclerosis. Furthermore, the exploration extends to the application of TCM monomers, elucidating their efficacy as therapeutic agents in these conditions. Additionally, the review examines the utilization of exosomes as drug delivery carriers in the context of neurodegenerative diseases, providing a nuanced understanding of the potential synergies between TCM and modern therapeutic approaches. This synthesis of knowledge aims to contribute to the advancement of our comprehension of the intricate molecular mechanisms underlying neurodegeneration and the potential therapeutic avenues offered by TCcom interventions.

Improved survival outcome with not-delayed radiotherapy and immediate PD-1/PD-L1 inhibitor for non-small-cell lung cancer patients with brain metastases.

PURPOSE: To investigate the impact of radiotherapy (RT) and immune checkpoint inhibitor (ICI) sequence on the survival outcome in NSCLC patients with brain metastasis, and decide the best time to initiate RT. METHODS: Patients were managed with delayed RT (ICI delivered over 2 weeks prior to RT), concurrent RT (ICI delivered within 2 weeks prior to or after RT), or upfront RT (RT delivered over 2 weeks prior to ICI). Overall survival (OS), intracranial local progression-free survival (iLPFS), and intracranial distant progression-free survival (iDPFS) were assessed. A meta-analysis was performed to analyze the association between survival outcome and RT/ICI sequence. RESULTS: A total of 73 NSCLC patients were identified with a median follow-up of 13.9 months. Patients who receive delayed RT demonstrated shorter iLPFS (P = 0.0029), iDPFS (P = 0.016), and OS (P < 0.001). A meta-analysis was conducted and a total of 4 studies, 254 patients were included. The HR was 0.44 for iDPFS (P = 0.03), 0.41 for OS (P < 0.01) when compared concurrent with delayed RT, 0.21 for iDPFS (P < 0.01), 0.32 for OS (P < 0.01) when compared upfront with delayed RT, consistent with our conclusion that delayed RT brought with worst iDPFS and OS. More importantly, the best overall response rate (BOR) decreased in cases with longer RT and ICI intervals. Patients who receive intervals of RT and ICI within 7 days achieve the best median BOR of - 53%. CONCLUSIONS: Delayed RT brought poor survival outcomes including iLPFS, iDPFS, and OS in NSCLC patients. The shorter interval of RT and ICI is associated with better BOR.

RNA Binding Protein RNPC1 Suppresses the Stemness of Human Endometrial Cancer Cells via Stabilizing MST1/2 mRNA.

BACKGROUND RNA binding protein RNPC1 has a tumor-suppressive role in various tumors, nevertheless, the role of RNPC1 in human endometrial cancer (EC) are never been reported. MATERIAL AND METHODS Western blot, quantitative polymerase chain reaction and sphere forming analysis were performed to evaluate the stem-like traits of cells and RNPC1-induced effects on EC cell stemness. RNA immunoprecipitation (RIP) was constructed to investigate the underlying mechanisms. RESULTS The spheres formed by EC cells, named EC spheres, exhibited a remarkably higher stemness than the parental cells, which is characterized as the increase of sphere forming ability, ALDH1 activity, stemness marker expression and migration ability. Notably, RNPC1 expression was decreased in poorly differentiated EC cells than that in EC cells with moderately differentiated. Additionally, RNPC1 expression was significantly decreased in EC spheres and RNPC1 overexpression attenuated the stemness of EC spheres. Moreover, RNPC1 overexpression decreased the migration ability of EC spheres. Mechanistic studies showed that RNPC1 overexpression activated the Hippo pathway through directly binding to MST1/2. Inhibition of MST1/2 rescued RNPC1-mediated effects on EC sphere stemness. CONCLUSIONS Therefore, our results indicate a novel RNPC1/MST1/2 signaling responsible for EC cell stemness.

The Long Non-Coding RNA MALAT1 Enhances Ovarian Cancer Cell Stemness by Inhibiting YAP Translocation from Nucleus to Cytoplasm.

BACKGROUND The purpose of this work was to unearth the effects and underlying mechanism of long non-coding RNA (lncRNA) MALAT1 in ovarian cancer cell stemness. MATERIAL AND METHODS Western blot, quantitative polymerase chain reaction (qPCR) and sphere forming analysis were performed to evaluate the stem-like traits of cells and MALAT1-induced effects on ovarian cancer cell stemness. Cell viability was performed to evaluate MALAT1 role in the chemoresistance of ovarian cancer cells. RNA immunoprecipitation (RIP) and luciferase reporter analysis were constructed to investigate the underlying mechanisms. RESULTS Here, qPCR assay showed that MALAT1 level was remarkably higher in non-adherent spheres formed by adherent ovarian cancer cells, as well as cisplatin-resistant ovarian cancer cells. Additionally, MALAT1 knockdown reduced ovarian cancer cell stemness, characterized as the decrease of sphere forming ability, expression of stemness regulatory masters, and attenuation of cisplatin resistance. Moreover, MALAT1 interacted with yes-associated protein (YAP), inhibited its nuclear-cytoplasm translocation, promoted YAP protein stability and expression and thus increased its activity. Notably, rescuing expression of YAP attenuated the inhibition of MALAT1 knockdown on ovarian cancer cell stemness. CONCLUSIONS In conclusion, these results demonstrate a MALAT1/YAP axis responsible for ovarian cancer cell stemness.

Tumor Necrosis Factor-α Polymorphisms and Cervical Cancer: Evidence from a Meta-Analysis.

BACKGROUND: Some previous studies already explored associations between tumor necrosis factor-α (TNF-α) polymorphisms and cervical cancer, with conflicting findings. OBJECTIVES: Here, we aimed to better analyze the relationship between TNF-α polymorphisms and cervical cancer in a larger combined population by performing a meta-analysis. METHODS: We searched PubMed, Embase, Web of Science, and CNKI for related articles. We calculated OR and 95% CI to estimate whether there are genetic associations between TNF-α polymorphisms and cervical cancer. RESULTS: Twenty-seven studies were included for this meta-analysis. TNF-α -308 G/A (dominant comparison: p = 0.0004, OR 0.71, 95% CI 0.58-0.86; recessive comparison: p = 0.0002, OR 1.46, 95% CI 1.19-1.79; overdominant comparison: p = 0.002, OR 1.37, 95% CI 1.12-1.68; allele comparison: p < 0.0001, OR 0.72, 95% CI 0.62-0.83) polymorphism was found to be significantly associated with cervical cancer in general population. Subgroup analyses showed similar positive findings for -308 G/A polymorphism in both Asians and Caucasians. Moreover, we found that -238 G/A polymorphism was also significantly associated with cervical cancer in Asians. CONCLUSIONS: This meta-analysis proved that TNF-α -238 and -308 G/A polymorphisms could be used to identity individual with elevated susceptibility to cervical cancer in certain populations.

The Wnt/ß-catenin signaling pathway affects the distribution of cytoskeletal proteins in Aß treated PC12 cells.

Alzheimer's disease is pathologically characterized by the presence of senile plaques and neurofibrillary tangles in the central nervous system. Amyloid ß-protein is toxic to neurons and induces phosphorylation of Tau protein, which accumulates in paired helical filaments and leads to the formation of neurofibrillary tangles. This study is focused on the Wnt/ß-catenin pathway influence on Tau phosphorylation and the distribution of microtubules and neurofilaments in adrenal pheochromocytoma cells. It was found that neurofilament heavy polypeptide and microtubule-associated protein-2 aggregated after treatment with Aß1₋42. Treatment with Wnt5a reduced this aggregation, while Dickkopf-1 treatment promoted microtubule and neurofilament aggregation. Furthermore, Tau phosphorylation at Ser396, Ser422, and Ser199 was significantly reduced after Wnt5a treatment, whereas Dickkopf-1 increased the level of phosphorylation. These results suggest that the Wnt/ß-catenin pathway influences the distribution of microtubules and neurofilaments, possibly by modulating the phosphorylation of Tau protein in adrenal pheochromocytoma cells.

Finite element simulation of treatment with locking plate for distal fibula fractures.

An improved Finite Element Model(FEM) is applied to compare the biomechanical stability of plates with three different options in the treatment of distal fibula fractures in this study. The Computed Tomography(CT) scan of the knee to ankle segment of a volunteer was performed. A 3D fibula FEM was reconstructed based on the CT data. Three different loads (uni-pedal standing, torsion, and twisting) were applied, the same as in the experiments in the literature. The stresses and strains of the three options were compared under the same loads, using a 4-hole locking plate (Option A), a 5-hole locking plate (Option B), and a 6-hole locking plate (Option C) in a standard plate for lateral internal fixation. The simulation results show that all three options showed a stress masking effect. Option C had the best overall biomechanical performance and could effectively distribute the transferred weight. This is because option C has greater torsional stiffness and better biomechanical stability than options A and B, and therefore, option C is the recommended internal fixation method for distal fibula fractures. The Finite Element Analysis(FEA) method developed in this work applies to the stress analysis of fracture treatment options in other body parts.

Higenamine exerts antidepressant effect by improving the astrocytic gap junctions and inflammatory response.

BACKGROUND: Depression is a refractory psychiatric disorder closely associated with dysfunction of the gap junctions (GJs) between astrocytes as well as neuroinflammation. Higenamine (Hig) is a potent cardiotonic ingredient in Fuzi (i.e., Aconitum carmichaeli Debx.) with anti-inflammatory and antioxidant effects, which has a significant protective effect on damaged nerve cells and has great potential for the treatment of neuropsychiatric diseases. METHODS: Rats were stimulated by chronic unpredictable stress (CUS) for 28 days while given Hig (5, 10, 20 mg/kg) and then analyzed behaviorally by the open field test, sucrose preference test, and forced swimming test. Changes in astrocyte GJs function and morphology were observed by dye transfer and transmission electron microscopy, respectively. Expression and phosphorylation of connexin 43 (Cx43) were analyzed by Western blot. Also, considering the close relationship between depression and neuroinflammation, we determined the inflammatory response in serum with ELISA kits and analyzed the expression of inflammation-related proteins with Western blot. RESULTS: Hig ameliorated CUS-induced depression-like behavior in rats. Hig administration improved gap junctional dysfunction in astrocytes, reduced gap junctional gaps and elevated the expression of Cx43 and decreased the phosphorylation of Cx43. Meanwhile, Hig administration was also able to attenuate the inflammatory response that occurs after CUS in rats. LIMITATIONS: For the role of Cx43 in depression, we did not validate it more deeply in animal models with knockout Cx43. In addition, GJs dysfunction might be associated with the inflammatory response seen in depression, but this needs to be further investigated. CONCLUSIONS: Hig ameliorates depression and exerts its antidepressant effect possibly by improving the dysfunctional GJs between astrocytes and the inflammatory response.

Microsurgical Anatomy of the Common Tendinous Ring and Its Surgical Implications.

BACKGROUND AND OBJECTIVES: The common tendinous ring (CTR), also known as the common annular tendon or annulus of Zinn, is a critical anatomic structure located at the convergence of the orbital apex, superior orbital fissure (SOF), optic canal, and the anterior aspect of the lateral sellar compartment. It plays a vital role in both neurosurgical and neuro-ophthalmological interventions. The aim of this study was to delineate the complex 3-dimensional (3D) topography of the CTR and explore its implications for surgical procedures. METHODS: Ten formalin-fixed skull base specimens from adult Chinese cadavers were meticulously dissected to investigate the morphology of the CTR, focusing particularly on its relationship with the 4 extraocular rectus tendons, the optic strut, the SOF, and the optic canal. Additional skull base specimens were subjected to 3D surface scanning, computed tomography, and histopathological examinations to deepen our understanding of the CTR's structural complexities. RESULTS: The CTR establishes a spatial, 3D tendinous assembly, encompassing 4 rectus tendons, 2 tendinous connections, and a singular common tendinous root. These components interlink to form a distinctive dual-ring configuration, featuring the optic foramen and the oculomotor foramen. The posterior part of the superior rectus tendon demarcates the common boundary between these 2 foramina. The oculomotor foramen itself serves as the central sector of the SOF. Precise incisions of the medial and lateral tendinous connections and fusions are essential for safely opening the CTR. CONCLUSION: The structural composition, interconnections, and dual-ring configuration of the CTR are crucial for precise and safe surgery of orbital apex and adjacent regions.

Effects of estrogen and phytoestrogens on endometrial leakage in ovariectomized rats and the related mechanisms.

Phytoestrogens, a group of plant-derived non-steroidal compounds that can behave as estrogens by binding to estrogen receptors, have drawn great attention for their potentially beneficial effects on human health. However, there are few studies investigating the potential side effects of phytoestrogens on the reproductive system. The present study was to elucidate the effects of 17ß-estradiol (E2), progesterone (P4), and phytoestrogens genistein (Gen), resveratrol (Res), and phloretin (Phl) on eosinophilic infiltration of the ovariectomized rat uterus and endometrial vascular permeability, and to analyze the underlying mechanisms. The ovariectomized rats received daily subcutaneous injections of E2, E2+P4, P4, Gen, Res, Phl, or an equivalent volume of vehicle for 21 days, and sham-operated animals (Sham rats) were used as the controls. Hematoxylin-eosin staining revealed a marked increase in uterine eosinophilic infiltrations in ovariectomized rats treated with E2, E2+P4 or P4, which was associated with increased expression of vascular endothelial growth factor (VEGF), nuclear factor-κB (NF-κB), and tumor necrosis factor-α (TNF-α) proteins as determined by immunohistochemical and Western blot analysis. However, all three phytoestrogens had no markedly effect on the uterine eosinophilic infiltration and the expressions of VEGF, NF-κB, and TNF-α in the uterus of ovariectomized rats. Our data demonstrate that E2 alone or in combination with P4 increases uterine eosinophilic infiltration which is related with vascular hyperpermeability caused by VEGF, NF-κB and TNF-α, whereas phytoestrogens Gen, Res, and Phl, have no such an effect.

The Formation and Phase Stability of A-Site High-Entropy Perovskite Oxides.

High entropy perovskite oxides (HEPOs) were a class of advanced ceramic materials, which had attracted much scientific attention in recent years. However, the effect of factors affecting the phase stability of high entropy perovskite oxides was still controversial. Herein, 17 kinds of A-site HEPOs were synthesized by solid-state methods, and several criteria for the formation of HEPOs and phase stability were investigated. Single-phase solid solutions were synthesized in 12 kinds of subsystems. The results show that the phase stability of a single-phase solid solution was affected by the size disorder and configurational entropy. The electronegativity difference was the key parameter to predict the evolution of the cubic/tetragonal phase, rather than the tolerance factor. Cubic HEPOs were easily formed when the electronegativity difference was <0.4, while the tetragonal HEPOs were easily formed when the electronegativity difference was ≥0.4. This study can further broaden the family of HEPOs and is expected to design the phase stability of HEPOs through electronegativity difference.

Novel insights into the progression and prognosis of the calpain family members in hepatocellular carcinoma: a comprehensive integrated analysis.

Objectives: The goal of our bioinformatics study was to comprehensively analyze the association between the whole calpain family members and the progression and prognosis of hepatocellular carcinoma (HCC). Methods: The data were collected from The Cancer Genome Atlas (TCGA). The landscape of the gene expression, copy number variation (CNV), mutation, and DNA methylation of calpain members were analyzed. Clustering analysis was performed to stratify the calpain-related groups. The least absolute shrinkage and selection operator (LASSO)-based Cox model was used to select hub survival genes. Results: We found 14 out of 16 calpain members expressed differently between tumor and normal tissues of HCC. The clustering analyses revealed high- and low-risk calpain groups which had prognostic difference. We found the high-risk calpain group had higher B cell infiltration and higher expression of immune checkpoint genes HAVCR2, PDCD1, and TIGHT. The CMap analysis found that the histone deacetylase (HDAC) inhibitor trichostatin A and the PI3K-AKT-mTOR pathway inhibitors LY-294002 and wortmannin might have a therapeutic effect on the high-risk calpain group. The DEGs between calpain groups were identified. Subsequent univariate Cox analysis of each DEG and LASSO-based Cox model obtained a calpain-related prognostic signature. The risk score model of this signature showed good ability to predict the overall survival of HCC patients in TCGA datasets and external validation datasets from the Gene Expression Omnibus database and the International Cancer Genome Consortium database. Conclusion: We found that calpain family members were associated with the progression, prognosis, and drug response of HCC. Our results require further studies to confirm.

Preoperative prediction of clinical and pathological stages for patients with esophageal cancer using PET/CT radiomics.

BACKGROUND: Preoperative stratification is critical for the management of patients with esophageal cancer (EC). To investigate the feasibility and accuracy of PET-CT-based radiomics in preoperative prediction of clinical and pathological stages for patients with EC. METHODS: Histologically confirmed 100 EC patients with preoperative PET-CT images were enrolled retrospectively and randomly divided into training and validation cohorts at a ratio of 7:3. The maximum relevance minimum redundancy (mRMR) was applied to select optimal radiomics features from PET, CT, and fused PET-CT images, respectively. Logistic regression (LR) was applied to classify the T stage (T1,2 vs. T3,4), lymph node metastasis (LNM) (LNM(-) vs. LNM(+)), and pathological state (pstage) (I-II vs. III-IV) with features from CT (CT_LR_Score), PET (PET_LR_Score), fused PET/CT (Fused_LR_Score), and combined CT and PET features (CT + PET_LR_Score), respectively. RESULTS: Seven, 10, and 7 CT features; 7, 8, and 7 PET features; and 3, 6, and 3 fused PET/CT features were selected using mRMR for the prediction of T stage, LNM, and pstage, respectively. The area under curves (AUCs) for T stage, LNM, and pstage prediction in the validation cohorts were 0.846, 0.756, 0.665, and 0.815; 0.769, 0.760, 0.665, and 0.824; and 0.727, 0.785, 0.689, and 0.837 for models of CT_LR_Score, PET_ LR_Score, Fused_ LR_Score, and CT + PET_ LR_Score, respectively. CONCLUSIONS: Accurate prediction ability was observed with combined PET and CT radiomics in the prediction of T stage, LNM, and pstage for EC patients. CRITICAL RELEVANCE STATEMENT: PET/CT radiomics is feasible and promising to stratify stages for esophageal cancer preoperatively. KEY POINTS: • PET-CT radiomics achieved the best performance for Node and pathological stage prediction. • CT radiomics achieved the best AUC for T stage prediction. • PET-CT radiomics is feasible and promising to stratify stages for EC preoperatively.

Effect of Current Density on the Microstructure and Mechanical Properties of 3YSZ/Al2O3 Composites by Flash Sintering.

The 3YSZ/40 wt% Al2O3 composites were prepared by flash sintering at a low furnace temperature (700 °C). The effects of the current density on the relative density and Vickers hardness of the composites were systematically investigated. The results showed that the relative densities and Vickers hardness of the samples increased gradually with the increasing of the current densities, and the relative density was as high as 94.2%. The Vickers hardness of 11.3 GPa was obtained under a current density of 102 mA/mm2. Joule heating and defects generation are suggested to be the main causes of rapid densification in flash sintering. The microstructure of the molten zone showed the formation of eutectic structures in the composite, suggesting that grain boundary overheating may have contributed to the formation of the molten zone.

Anisotropic Piezoelectric Properties of Porous (Ba0.85Ca0.15)(Zr0.1Ti0.9)O3 Ceramics with Oriented Pores through TBA-Based Freeze-Casting Method.

Porous (Ba0.85Ca0.15)(Zr0.1Ti0.9)O3 (BCZT) piezoelectric ceramics with an oriented directional hole structure were prepared by using the tertbutyl alcohol (TBA)-based freeze-casting method. The influences of sintering temperatures on the microstructure and piezoelectric properties of porous BCZT ceramics were investigated both perpendicular and parallel to the freezing direction. With the increase in sintering temperatures and the porosities decreased from 58% to 42%, the compressive strength increased from 14.0 MPa to 25.0 MPa. In addition, the d33 value of 407 pC/N for the sample sintered at 1400 °C was obtained parallel to the freezing direction, which was 1.40 times that of the other direction.

Implications of m6A-associated snRNAs in the prognosis and immunotherapeutic responses of hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) is the most prevalent pathological type of liver cancer worldwide with high mortality and poor prognosis. N6-methyladenosine (m6A) can modify RNAs such as mRNA, lncRNA, miRNA, and tRNA, thereby playing a critical role in the pathogenesis of HCC. However, the role of m6A-associated small nuclear RNA (snRNA) in the prognostic value and immunotherapeutic response in HCC remains unclear. Materials and methods: In this study, snRNA expression data, gene mutation data, and clinical data of HCC patients were acquired from The Cancer Genome Atlas (TCGA) database. We used the least absolute shrinkage and selection operator (LASSO) Cox regression analysis to identify significant prognostic m6A-associated snRNAs, and then developed a multivariate Cox model based on the selected snRNAs. HCC patients were split into low- and high-risk groups based on the median risk score. We subsequently performed Kaplan-Meier curve analysis to estimate overall survival (OS) by clinicopathological characteristics and tumor mutational burden (TMB) status in low- and high-risk HCC patients. Finally, we compared the immunotherapeutic response as represented by tumor immune dysfunction and exclusion (TIDE) scores between the two risk groups. Results: Eight m6A-associated snRNAs were selected as independent predictors to develop the risk model. Our results revealed that the OS of HCC patients in the high-risk group was significantly worse than that in the low-risk group on clinicopathologic characteristics, including age (≤65 years and >65 years), gender (male), grade (G I-II and G III-IV) and TNM staging (Stage I-II and Stage III-IV). In addition, the OS of low-TMB and low-risk group was longer than that of high-TMB and high-risk group. The TIDE score indicated that HCC patients in the high-risk group were more susceptible to immunotherapy. Conclusion: Our study suggests that m6A-associated snRNAs may be useful biomarkers for the prognosis of HCC and that m6A-associated snRNA models can predict the effect of immunotherapy in HCC patients.

Icotinib With Concurrent Radiotherapy vs Radiotherapy Alone in Older Adults With Unresectable Esophageal Squamous Cell Carcinoma: A Phase II Randomized Clinical Trial.

Importance: Palliative radiotherapy (RT) is generally recommended for older patients with esophageal squamous cell carcinoma (ESCC) with poor prognosis. A new combination treatment is therefore needed. Objective: To assess the efficacy and toxicity of RT plus icotinib vs RT alone in older patients with ESCC. Design, Setting, and Participants: This randomized, multicenter, open-label, phase II clinical trial was conducted in China, with enrollment between January 1, 2015, and October 31, 2016. Patients aged 70 years or older with clinical stage T2 to T4, N0/1, M0/1a unresectable (because of comorbidities, T4 disease, unresectable lymph node, or refused surgery) ESCC were randomized 1:1 to receive RT plus icotinib or RT alone. Radiation was prescribed at 60 Gy in 30 fractions in both groups, and icotinib was administered at a dosage of 125 mg 3 times a day in the RT plus icotinib group. The last follow-up was completed on June 30, 2019, and data were analyzed from July 1 to September 30, 2019. Interventions: Patients were randomized to either RT plus icotinib or RT alone. Main Outcomes and Measures: The primary end point was overall survival (OS). Treatment-related toxic effects were evaluated. Immunohistochemistry was performed to analyze epidermal growth factor receptor (EGFR) expression if available. Results: A total of 127 patients (median age, 76 years [range, 70-91 years]; 76 men [59.8%]) were enrolled and were eligible for survival analysis. Median OS was 24.0 (95% CI, 22.2-25.8) months in the RT plus icotinib group vs 16.3 (95% CI, 13.8-18.8) months in the RT group (hazard ratio, 0.53; 95% CI, 0.33-0.87; P = .008). No difference was observed in grades 3 or 4 adverse events. Patients with EGFR overexpression had a significantly better median overall survival (not reached vs 16.3 months [range, 2.6-45.1 months]; P = .03) in the RT plus icotinib group. Conclusions and Relevance: In this randomized clinical trial, icotinib plus RT was well tolerated and improved OS in older patients with ESCC relative to RT alone. Patients with EGFR overexpression benefitted more from icotinib with RT. Trial Registration: ClinicalTrials.gov Identifier: NCT02375581.

Genistein stimulates myocardial contractility in guinea pigs by different subcellular mechanisms.

The purpose of this study was to investigate the mechanisms involved in the excitatory effect induced by genistein in isolated guinea pig left ventricular papillary muscles and to determine relationship of genistein action with the tyrosine kinase pathway and phosphatidylinositol 3-kinase (PI3K) activity, the cyclic adenosine 5'-monophosphate (cAMP) signal system and the sarcoplasmic reticulum Ca2+ mobilization. Genistein (1-100 microM) significantly increased contraction of left ventricular papillary muscles from male and female guinea pigs in a concentration-dependent manner and its action had no obvious gender differences. Prior treatment with an L-type Ca2+ channel blocker verapamil hydrochloride, beta-adrenoceptor inhibitors propranolol and atenolol, an inhibitor of Na+-Ca2+ reverse exchanger Kb-r7943 or the blocker of estrogen receptor ICI 182,780 failed to alter the positive inotropic effect induced by genistein in papillary muscles. However, tyrosine phosphatase inhibitor, sodium orthovanadate or a potent phosphotyrosine phosphatase inhibitor bpV (phen) could partly but significantly reduce the stimulatory action of genistein. Interestingly, insulin-like growth factor-1, a known PI3K activator could also decrease the stimulatory action of genistein obviously, but the PI3K inhibitor LY294002 had no significant effect on the stimulatory action of genistein. The excitatory effect of genistein was markedly attenuated not only after treatment with an inhibitor of cAMP synthesis Sq 22536, carbachol or an inhibitor of specific protein kinase A H-89, but also after the inhibition of sarcoplasmic reticulum Ca2+ mobilization by ruthenium red, ryanodine or the inhibitor of sarcoplasmic reticulum Ca2+-ATPase thapsigargin. All these results indicate that the excitatory effects of genistein in papillary muscles are due to the inhibition of tyrosine kinase pathway and PI3K activity, thereby locally activating cAMP signal transduction and facilitating intracellular Ca2+ mobilization, but are not related to the activation of beta-adrenoceptor, the Na+-Ca2+ reverse exchange and the estrogen receptor.

Inhibitory effects of genistein and resveratrol on guinea pig gallbladder contractility in vitro.

AIM: To observe and compare the effects of phytoestrogen genistein, resveratrol and 17beta-estradiol on the tonic contraction and the phasic contraction of isolated gallbladder muscle strips and to study the underlying mechanisms. METHODS: Isolated strips of gallbladder muscle from guinea pigs were suspended in organ baths containing Kreb's solution, and the contractilities of strips were measured before and after incubation with genistein, resveratrol and 17beta-estradiol respectively. RESULTS: Similar to 17beta-estradiol, genistein and resveratrol could dose-dependently inhibit the phasic contractile activities, they decreased the mean contractile amplitude and the contractile frequencies of gallbladder muscle strips, and also produced a marked reduction in resting tone. The blocker of estrogen receptor ICI 182780 failed to alter the inhibitory effects induced by genistein and resveratrol, but potassium bisperoxo (1, 10 phenanthroline) oxovanadate bpV (phen), a potent protein tyrosine phosphatase inhibitor, markedly attenuated the inhibitory effects induced by genistein and resveratrol. In calcium-free Kreb's solution containing 0.01 mmol/L egtazic acid (EGTA), genistein and resveratrol inhibited the first phasic contraction induced by acetylcholine (ACh), but did not affect the second contraction induced by CaCl(2). In addition, genistein, resveratrol and 17beta-estradiol also could reduce the contractile responses of ACh and KCl, and shift their cumulative concentration-response curves rightward. CONCLUSION: Phytoestrogen genistein and resveratrol can directly inhibit the contractile activity of isolated gallbladder muscle both at rest and in response to stimulation. The mechanisms responsible for the inhibitory effects probably due mainly to inhibition of tyrosine kinase, Ca(2+) influx through potential-dependent calcium channels (PDCs) and Ca(2+) release from sarcoplasmic reticulum (SR), but were not related to the estrogen receptors.

Randomised phase III trial of concurrent chemoradiotherapy with extended nodal irradiation and erlotinib in patients with inoperable oesophageal squamous cell cancer.

BACKGROUND: This randomised phase III study was conducted to investigate the efficacy of extended nodal irradiation (ENI) and/or erlotinib in inoperable oesophageal squamous cell cancer (ESCC). PATIENTS AND METHODS: Patients with histologically confirmed locally advanced ESCC or medically inoperable disease were randomly assigned (ratio 1:1:1:1) to one of four treatment groups: group A, radiotherapy adoption of ENI with two cycles of concurrent TP chemotherapy (paclitaxel 135 mg/m2 day 1 and cisplatin 20 mg/m2 days 1-3, every 4 weeks) plus erlotinib (150 mg per day during chemoradiotherapy); group B, radiotherapy adoption of ENI with two cycles of concurrent TP; group C, radiotherapy adoption of conventional field irradiation (CFI) with two cycles of concurrent TP plus erlotinib; group D, radiotherapy adoption of CFI with two cycles of concurrent TP. RESULTS: A total of 352 patients (88 assigned to each treatment group) were enrolled. The 2-year overall survival rates of group A, B, C and D were 57.8%, 49.9%, 44.9% and 38.7%, respectively (P = 0.015). Group A significantly improved 2-year overall survival compared with group D. The ENI significantly improved overall survival in patients with inoperable ESCC (P = 0.014). The addition of erlotinib significantly decreased loco-regional recurrence (P = 0.042). Aside from rash and radiation oesophagitis, the incidence of grade 3 or greater toxicities did not differ among 4 groups. CONCLUSION: Chemoradiotherapy with ENI and erlotinib might represent a substantial improvement on the standard of care for inoperable ESCC. ENI alone should be adopted in concurrent chemoradiotherapy for ESCC patients.