QM/MM Simulations of Afatinib-EGFR Addition: The Role of ß-Dimethylaminomethyl Substitution.

Acrylamides are the most commonly used warheads of targeted covalent inhibitors (TCIs) directed at cysteines; however, the reaction mechanisms of acrylamides in proteins remain controversial, particularly for those involving protonated or unreactive cysteines. Using the combined semiempirical quantum mechanics (QM)/molecular mechanics (MM) free energy simulations, we investigated the reaction between afatinib, the first TCI drug for cancer treatment, and Cys797 in the EGFR kinase. Afatinib contains a ß-dimethylaminomethyl (ß-DMAM) substitution which has been shown to enhance the intrinsic reactivity and potency against EGFR for related inhibitors. Two hypothesized reaction mechanisms were tested. Our data suggest that Cys797 becomes deprotonated in the presence of afatinib, and the reaction proceeds via a classical Michael addition mechanism, with Asp800 stabilizing the ion-pair reactant state ß-DMAM+/C797- and the transition state of the nucleophilic attack. Our work elucidates an important structure-activity relationship of acrylamides in proteins.

QM/MM simulations of EFGR with afatinib reveal the role of the ß-dimethylaminomethyl substitution.

Acrylamides are the most commonly used warheads of targeted covalent inhibitors (TCIs) directed at cysteines; however, the reaction mechanisms of acrylamides in proteins remain controversial, particularly for those involving protonated or unreactive cysteines. Using the combined semiempirical quantum mechanics (QM)/molecular mechanics (MM) free energy simulations, we investigated the reaction between afatinib, the first TCI drug for cancer treatment, and Cys797 in the EGFR kinase. Afatinib contains a ß-dimethylaminomethyl (ß-DMAM) substitution which has been shown to enhance the intrinsic reactivity and potency against EGFR for related inhibitors. Two hypothesized reaction mechanisms were tested. Our data suggest that Cys797 becomes deprotonated in the presence of afatinib and the reaction proceeds via a classical Michael addition mechanism, with Asp800 stabilizing the ion-pair reactant state ß-DMAM+/C797- and the transition state of the nucleophilic attack. Our work elucidates an important structure-activity relationship of acrylamides in proteins.

Developing a Novel Enzalutamide-Resistant Prostate Cancer Model via AR F877L Mutation in LNCaP Cells.

Prostate cancer is a leading diagnosis and major cause of cancer-related deaths in men worldwide. As a typical hormone-responsive disease, prostate cancer is commonly managed with androgen deprivation therapy (ADT) to curb its progression and potential metastasis. Unfortunately, progression to castration-resistant prostate cancer (CRPC), a notably more aggressive phase of the disease, occurs within a timeframe of 2-3 years following ADT. Enzalutamide, a recognized androgen receptor (AR) antagonist, has been employed as a standard of care for men with metastatic castration-resistant prostate cancer (mCRPC) since it was first approved in 2012, due to its ability to prolong survival. However, scientific evidence suggests that sustained treatment with AR antagonists may induce acquired AR mutations or splice variants, such as AR F877L, T878A, and H875Y, leading to drug resistance and thereby diminishing the therapeutic efficacy of these agents. Thus, the establishment of prostate cancer models incorporating these particular mutations is essential for developing new therapeutic strategies to overcome such resistance and evaluate the efficacy of next-generation AR-targeting drugs. We have developed a CRISPR (clustered regularly interspaced short palindromic repeats)-based knock-in technology to introduce an additional F877L mutation in AR into the human prostate cell line LNCaP. This article provides comprehensive descriptions of the methodologies for cellular gene editing and establishment of an in vivo model. Using these methods, we successfully identified an enzalutamide-resistant phenotype in both in vitro and in vivo models. We also assessed the efficacy of target protein degraders (TPDs), such as ARV-110 and ARV-667, in both models, and the corresponding validation data are also included here. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Generation of AR F877L-mutated LNCaP cell line using CRISPR technology Basic Protocol 2: Validation of drug resistance in AR F877L-mutated LNCaP cell line using the 2D CTG assay Support Protocol: Testing of sgRNA efficiency in HEK 293 cells Basic Protocol 3: Validation of drug resistance in AR F877L-mutated LNCaP cell line in vivo.

Pilose antler extract promotes hair growth in androgenic alopecia mice by promoting the initial anagen phase.

Androgenetic alopecia (AGA) is a prevalent disease in worldwide, local application or oral are often used to treat AGA, however, effective treatments for AGA are currently limited. In this work, we observed the promoting the initial anagen phase effect of pilose antler extract (PAE) on hair regeneration in AGA mice. We found that PAE accelerated hair growth and increased the degree of skin blackness by non-invasive in vivo methods including camera, optical coherence tomography and dermoscopy. Meanwhile, HE staining of sagittal and coronal skin sections revealed that PAE augmented the quantity and length of hair follicles, while also enhancing skin thickness and hair papilla diameter. Furthermore, PAE facilitated the shift of the growth cycle from the telogen to the anagen phase and expedited the proliferation of hair follicle stem cells and matrix cells in mice with AGA. This acceleration enabled the hair follicles to enter the growth phase at an earlier stage. PAE upregulated the expression of the sonic hedgehog (SHH), smoothened receptor, glioma-associated hemolog1 (GLI1), and downregulated the expression of bone morphogenetic protein 4 (BMP4), recombinant mothers against decapentaplegic homolog (Smad) 1 and 5 phosphorylation. This evidence suggests that PAE fosters hair growth and facilitates the transition of the growth cycle from the telogen to the anagen phase in AGA mice. This effect is achieved by enhancing the proliferation of follicle stem cells and matrix cells through the activation of the SHH/GLI pathway and suppression of the BMP/Smad pathway.

Quantum Descriptors for Predicting and Understanding the Structure-Activity Relationships of Michael Acceptor Warheads.

Predictive modeling and understanding of chemical warhead reactivities have the potential to accelerate targeted covalent drug discovery. Recently, the carbanion formation free energies as well as other ground-state electronic properties from density functional theory (DFT) calculations have been proposed as predictors of glutathione reactivities of Michael acceptors; however, no clear consensus exists. By profiling the thiol-Michael reactions of a diverse set of singly- and doubly-activated olefins, including several model warheads related to afatinib, here we reexamined the question of whether low-cost electronic properties can be used as predictors of reaction barriers. The electronic properties related to the carbanion intermediate were found to be strong predictors, e.g., the change in the Cß charge accompanying carbanion formation. The least expensive reactant-only properties, the electrophilicity index, and the Cß charge also show strong rank correlations, suggesting their utility as quantum descriptors. A second objective of the work is to clarify the effect of the ß-dimethylaminomethyl (DMAM) substitution, which is incorporated in the warheads of several FDA-approved covalent drugs. Our data suggest that the ß-DMAM substitution is cationic at neutral pH in solution and promotes acrylamide's intrinsic reactivity by enhancing the charge accumulation at Cα upon carbanion formation. In contrast, the inductive effect of the ß-trimethylaminomethyl substitution is diminished due to steric hindrance. Together, these results reconcile the current views of the intrinsic reactivities of acrylamides and contribute to large-scale predictive modeling and an understanding of the structure-activity relationships of Michael acceptors for rational TCI design.

Catalytic Mechanism of Human T-Cell Leukemia Virus Type 1 Protease Investigated by Combined QM/MM Molecular Dynamics Simulations.

Combined quantum mechanical and molecular mechanical (QM/MM) molecular dynamics simulations were performed to investigate the catalytic mechanism of human T-cell leukemia virus type 1 (HTLV-1) protease, a retroviral aspartic protease that is a potential therapeutic target for curing HTLV-1-associated diseases. To elucidate the proteolytic cleavage mechanism, we determined the two-dimensional free energy surfaces of the HTLV-1 protease-catalyzed reactions through various possible pathways. The free energy simulations suggest that the catalytic reactions of the HTLV-1 protease occur in the following sequential steps: (1) a proton is transferred from the lytic water to Asp32', followed by the nucleophilic addition of the resulting hydroxyl to the carbonyl carbon of the scissile bond, forming a tetrahedral oxyanion intermediate, and (2) a proton is transferred from Asp32 to the peptide nitrogen of the scissile bond, leading to the spontaneous breakage of the scissile bond. The rate-limiting step of this catalytic process is the proton transfer from Asp32 to the peptide nitrogen of the scissile bond, with a free energy of activation of 21.1 kcal/mol. This free energy barrier is close to the experimentally determined free energy of activation (16.3 kcal/mol) calculated from the measured catalytic rate constant (kcat). This mechanistic study provides detailed dynamic and structural information that will facilitate the design of mechanism-based inhibitors for the treatment of HTLV-1-associated diseases.

Ultrasonic breast tumor extraction based on adversarial mechanism and active contour.

BACKGROUND AND OBJECTIVE: Breast cancer is a high incidence of gynecological diseases; breast ultrasound screening can effectively reduce the mortality rate of breast cancer. In breast ultrasound images, the localization and segmentation of tumor lesions are important steps for the extraction of lesions, which helps clinicians evaluate breast lesions quantitatively and makes better clinical diagnosis of the disease. However, the segmentation of breast lesions is difficult due to the blurred and uneven edges of some lesions. In this paper, we propose a segmentation framework combining active contour module and deep learning adversarial mechanism and apply it for the segmentation of breast tumor lesions. METHOD: We use a conditional adversarial network as the main framework. The generator is a segmentation network consisting of a Deformed U-Net and an active contour module. Here, the Deformed U-Net performs pixel-level segmentation for breast ultrasound images. The active contour module refines the tumor lesion edges, and the refined result provides loss information for Deformed U-Net. Therefore, the Deformed U-Net can better classify the edge pixels. The discriminator is the Markov discriminator; this discriminator provides loss feedback for the segmentation network. We cross-train the discriminator and segmentation network to implement Adversarial Mechanism for getting a more optimized segmentation network. RESULTS: The segmentation performance of the segmentation network for breast ultrasound images is improved by adding a Markov discriminator to provide discriminant loss training. The proposed method for segmenting the tumor lesions in breast ultrasound image obtains dice coefficient: 89.7%, accuracy: 98.1%, precision: 86.3%, mean-intersection-over-union: 82.2%, recall: 94.7%, specificity: 98.5% and F1score: 89.7%. CONCLUSION: Comparing with traditional methods, the proposed method gives better performance. The experimental results show that the proposed method can effectively segment the lesions in breast ultrasound images, and then assist doctors to realize the diagnosis of breast lesions.

Risk factors and predictive model for abdominal wound dehiscence in neonates: a retrospective cohort study.

BACKGROUND: Abdominal wound dehiscence (AWD) is a major complication of abdominal surgery, and neonates are a group with a high risk of AWD, which has serious consequences or can even result in death. The purpose of this study is to explore the risk factors for neonatal AWD and construct a predictive model. METHODS: The clinical data of 453 cases that underwent neonatal laparotomy from June 2009 to June 2020 were retrospectively analyzed, among which 27 cases of AWD were identified. Nine factors, including gender, age at admission, weight at admission, preterm delivery, level of preoperative anaemia, hypoalbuminemia, operation time, incision length, and incision type, were analyzed to explore their correlation with neonatal AWD. RESULTS: The incidence of neonatal AWD was 6.0% (27/453), among which partial wound dehiscence accounted for 4.9% (22/453) and complete wound dehiscence accounted for 1.1% (5/453). Hypoproteinemia and incision type were the independent risk factors for neonatal AWD, and weight at admission was a protective factor for AWD in the multivariate models. All these factors were incorporated to construct a nomogram, and a calibration curve was plotted. The result indicated that the actual risk was close to the predicted risk when the predicted risk rate was greater than about 35%. CONCLUSIONS: Neonatal AWD is closely related to hypoproteinemia and incision contamination. Our predictive model showed the potential to provide an individualized risk estimate of AWD for neonatal patients undergoing abdominal surgery.Key messagesNeonatal abdominal wound dehiscence (AWD) has a serious consequence and the incidence of neonatal AWD was about 6.0% and the complete AWD morbidity is 1.1%.Hypoproteinemia and incision type were the independent risk factors for neonatal AWD.Our predictive model showed the potential to provide an individualized risk estimate of AWD for neonatal patients undergoing abdominal surgery.

Noni (Morinda citrifolia L.) wine prevents the oxidative stress and obesity in mice induced by high-fat diet.

Noni (Morinda citrifolia L.) is rich in polyphenols, flavonoids, terpenoids, and iridoids. However, its bad taste and smell make noni fruit unsuitable for consumption. After fermentation, noni wine becomes free from the undesirable smell. Nevertheless, it is still unclear whether processed noni could retain its original nutrients and effects. Therefore, we conducted a series of evaluations on the nutritional composition and efficacy of noni wine. Our results showed that the polyphenol, flavonoid, and vitamin C contents in noni wine were 558.80, 234.42, and 0.30 mg/L, respectively. Our animal experiments showed that 40 ml kg-1  day-1 noni wine could reduce bodyweight, as well as the levels of body fat, serum triglycerides, total cholesterol, and low-density lipoprotein, while it simultaneously increased the amount of energy expenditure and activity, and improved the systemic antioxidant capacity in mice following a high-fat diet. The results of the gene expression and western blot analyses showed that 40 ml kg-1  day-1 noni wine could regulate the Nrf2 pathway and improve the antioxidant enzyme gene expression in mice maintained on a high-fat diet, thereby improving body lipid metabolism, reducing fatty acid synthesis, and promoting fatty acid ß-oxidation. Our study indicated that drinking 40 ml kg-1  day-1 noni wine could effectively prevent high-fat diet-induced oxidative stress and obesity in mice. PRACTICAL APPLICATIONS: Noni fruit is rich in nutrients but its bad smell and hardship of processing make its commercialization difficult. Previous studies mainly focused on fresh noni juice and its primary processed products, while few noni products, of poor taste and low quality, are available in the market. Therefore, the fruit wine with both the nutritive values and the special flavor of noni has broad market prospects. Our work provides a valuable reference for the commercialization of noni wine.

Evaluating the Activity of Sodium Butyrate to Prevent Osteoporosis in Rats by Promoting Osteal GSK-3ß/Nrf2 Signaling and Mitochondrial Function.

Oxidative stress (OS) and mitochondrial dysfunction are key pathophysiological features of osteoporosis and obesity. Sodium butyrate (NaB), produced by fermentation by the gut microbiota of the large intestine, has been demonstrated to protect against OS by improving specific antioxidant enzymes and to regulate mitochondria redox homeostasis in vivo. Here, in an unblinded study, we identified femur mitochondria as the main target of the beneficial effects of NaB, consisting of reversion of bone loss and body-weight gain in obesity-prone rats. In particular, NaB promoted the activity of mitochondrial antioxidant enzymes and energy metabolism, preserved the bone microstructure and calcium homeostasis, and activated bone metabolism, as shown by increased Nrf2/GSK-3ß signaling and the upregulation of PGC-1α and TFAM. In vitro experiments showed that moderate NaB treatment prevented H2O2-induced oxidative damage in MC3T3-E1 cells, improved osteoblast mineralization and differentiation, and maintained the balance in bone metabolism by enhancing intracellular antioxidant enzyme activity and ATP production and decreasing the ROS level. In conclusion, NaB promoted the Nrf2/GSK-3ß signaling pathway and mitochondrial function and is a potential new therapeutic strategy for obesity and osteoporosis.

Effects of Chronic Ephedrine Toxicity on Functional Connections, Cell Apoptosis, and CREB-Related Proteins in the Prefrontal Cortex of Rhesus Monkeys.

Ephedrine abuse has spread in many parts of the world, severely threatening human health. The mechanism of ephedrine toxicity is still unclear. To explore the possible neural mechanisms of ephedrine toxicity, this study established a non-human primate model of ephedrine exposure, analyzed the functional connectivity changes in its prefrontal cortex through resting state BOLD-fMRI, and then inspected the pathophysiological changes as well as the expression of the cyclic adenosine monophosphate response element-binding protein (CREB), phosphorylated CREB (P-CREB), and CREB target proteins (c-fos and fosB) in the prefrontal cortex. After ephedrine toxicity, we found that the prefrontal cortex of monkeys strengthened its functional connectivity with the brain regions that perform motivation, drive, reward, and learning and memory functions and weakened its functional connectivity with the brain regions that perform cognitive control. These results suggest that ephedrine toxicity causes abnormal neural circuits that lead to the amplification and enhancement of drug-related cues and the weakening and damage of cognitive control function. Histology showed that the neurocytotoxicity of ephedrine can cause neuronal degeneration and apoptosis. Real-time PCR and Western blot showed increased expression of CREB mRNA and CREB/P-CREB/c-fos/fosB protein in the prefrontal cortex after ephedrine toxicity. Collectively, the present study indicates that the enhancement of drug-related cues and the weakening of cognitive control caused by abnormal neural circuits after drug exposure may be a major mechanism of brain function changes caused by ephedrine. These histological and molecular changes may be the pathophysiological basis of brain function changes caused by ephedrine.

Pilose Antler Extracts (PAEs) Protect against Neurodegeneration in 6-OHDA-Induced Parkinson's Disease Rat Models.

Parkinson's disease (PD) is one of the most common neurodegenerative diseases worldwide. Although dopamine replacement therapy mitigates motor dysfunction in PD patients, there are no therapeutics that are currently available to reverse neuronal cell death in the substantia nigra pars compacta (SNc), which is the main region for dopamine loss in PD patients. The protein concentration of the Pilose antler extracts (PAEs) was estimated using the Bradford Protein Assay Kit. Hematoxylin and eosin (HE) staining was used to evaluate the protective effect of PAEs on 6-OHDA induced cell death in PD model rats. Immunohistochemistry (IHC) was used to detect the tyrosine hydroxylase (TH) positive neuronal cell in SNc. HPLC-MS was used to detect dopamine (DA), 3,4-Dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), and glutamate (Glu) levels in the striatum and cerebrospinal fluid (CSF). The amino acid level in the striatum and CSF was measured by HPLC-FLD. Protein expression of growth associated protein-43 (GAP-43) and neurofilament heavy polypeptide (NF-H) was measured using western blotting. The components of PAEs through blood vessels were detected by HPLC/MS/MS. In this study, PAEs with proteins ranging from 10 kDa to 250 kDa molecular weight was administered to 6-OHDA-induced PD rats. We found that PAEs inhibited 6-OHDA-induced neuronal cell death and TH-positive neuronal loss in SNc. PAEs administration also increased the levels of DA, DOPAC, and 5-HT, in addition to DOPAC/DA and HVA/DA indexes in the CSF and Striatum of 6-OHDA induced rats. Conversely, PAEs decreased the levels of Glu and GABA. Treatment with PAEs and Madopar increased GAP-43 and NF-H expression in the SNc and striatum. Proteomic analysis using LC/MS/MS indicated that 11 components of PAEs may have neuropharmacological effects. These results demonstrate that PAEs protects against 6-OHDA induced toxic effects in the PD rat models. Intragastric administration of PAEs may be a novel therapeutic strategy for neurodegenerative disorders like PD.

Megameatus intact prepuce treated with urethral plate-preserving surgery: a retrospective study of an unusual hypospadias variant.

BACKGROUND: Megameatus intact prepuce (MIP) is a unique variant of hypospadias and is a clinically rare condition. Due to the anatomical characteristics of the MIP hypospadias variant presenting a unique challenge to surgeons, no single urethroplasty method provides a universal solution for all patients. The purpose of this study was to evaluate the outcomes of hypospadias after MIP repair by urethral plate-preserving urethroplasty. METHODS: A retrospective study was performed on 25 coronal or distal MIP patients, with a median age of 8, with most deficiencies being discovered during their first hospital visit for phimosis. Correction with urethroplasty was performed for all patients; 5 underwent the Mathieu procedure, 13 underwent the tubularized incised plate (TIP) procedure, and 7 underwent the Duplay procedure. The 25 patients were followed up for 6 to 36 months to evaluate the surgical outcomes. RESULTS: There were no significant differences in intraoperative bleeding, hospital stays, postoperative analgesia rate, and cure rate among the three surgical procedures. The operative time for the Mathieu procedure was longer than that for the TIP and Duplay procedures, which did not differ. Complications occurred in 4 of the 25 patients (16.0%), and the overall complication-free survival rate at 1 year after surgery was 80.5%. The age at the time of surgery, urethral plate width, urethroplasty length, surgical procedures, or meatal location (coronal or distal penis) were not independently predictive of complications. CONCLUSIONS: The clinical manifestations of MIP are often concealed and then accidentally discovered during hospital visits for phimosis; thus, the actual incidence of MIP might be higher. The urethral plate should be preserved during MIP-correcting treatment, especially for coronal or distal MIP. The same satisfactory outcomes can be obtained with Mathieu, TIP, or Duplay urethroplasty.

Nicotinic Acid Receptor GPR109A Exerts Anti-Inflammatory Effects Through Inhibiting the Akt/mTOR Signaling Pathway in MIN6 Pancreatic ß cells.

OBJECTIVES: We found that activation of the nicotinic acid receptor GPR109A, expressed by the MIN6 murine pancreatic ß cell line, inhibits nitric oxide accumulation induced by IFN-γ and TNF-α, implicating an anti-inflammatory effect of GPR109A in MIN6 cells. Nevertheless, the mechanism of its anti-inflammatory effect is still unknown. In this study, we used palmitic acid to stimulate MIN6 cells to induce inflammatory cytokine production and explored the mechanism by which GPR109A exerts anti-inflammatory effects. MATERIALS AND METHODS: RT-PCR and immunocytochemical staining were used to detect the expression of GPR109A in MIN6 cells. Western blotting was used to detect the activation of the Akt/mTOR signaling pathway and expression of the inflammatory cytokine INF-γ, in MIN6 cells, following treatments with palmitic acid and palmitic acid+nicotinic acid, or with different concentrations of nicotinic acid and 3-hydroxybutyrate. RESULTS: In MIN6 cells, GPR109A transcripts and protein are expressed and GPR109A protein is mainly located in the cell membrane and cytoplasm. Palmitic acid enhanced the phosphorylation of Akt and p70S6K and elevated the expression of IFN-γ. Co-treatment with nicotinic acid, which is an agonist of GPR109A, inhibited the palmitic acid-induced phosphorylation of Akt, mTOR, and p70S6K, as well as the expression of IFN-γ. CONCLUSIONS: GPR109A may inhibit inflammatory cytokine production, induced by palmitic acid, by MIN6 cells possibly via inhibiting the Akt/mTOR signaling pathway.

Peutz-Jeghers syndrome with intermittent upper intestinal obstruction: A case report and review of the literature.

RATIONALE: Peutz-Jeghers syndrome (PJS) is an autosomal dominant genetic syndrome characterized by a unique type of gastrointestinal hamartomatous polyp associated with oral and anal mucocutaneous pigmentations. Peutz-Jeghers polyps occur most numerously in the small intestine but frequently in the colon and stomach, only a few cases have been reported in the duodenum. PATIENT CONCERN: A further family history survey discovered 10 out of 14 members of the family (in 4 generations) had mucocutaneous pigmentations, but many of them were living in rural areas where they had no access to specialized medical services, so none were checked with endoscopy for polyps of hamartoma. DIAGNOSES: We report the case of a boy patient with mucocutaneous pigmentations over the lips, and a history of recurrent bouts of vomit and anemia over the preceding two years, no abdominal pain and mass. An upper gastrointestinal endoscopy revealed some small polyps in the stomach and multiple sessile polyps in the second part of the duodenum, but colonoscopy exam did not reveal any lesion. INTERVENTIONS: A double polypectomy and duodenum segmentary resection with end-to-end anastomosis was performed. Histopathology of the resected duodenum polyps indicated it to be a typical hamartomatous polyp. OUTCOMES: The child was under regular follow-up and recovered well. LESSONS: In this case, the patient was characteristic with pigmentations on his lips and intermittent upper intestinal obstruction (due to mass duodenal polyps), there are no definitive guidelines for the treatment to duodenal PJS hamartomatous polyp, each case requires tailor-made management.

Velvet antler polypeptide is able to induce differentiation of neural stem cells towards neurons in vitro.

OBJECTIVE: To investigate the neural differentiation capacity of water extraction of velvet antler. METHODS: Velvet antler (Cervus Nippon Temminck) polypeptide (VAP) was used to differentiate neural stem cells (NSCs) towards neurons in the study. Firstly, we obtain the polypeptides of VAP by water extraction. Secondly, we observed the morphology, assayed the factors in the media by enzyme-linked immunosorbent assay, and detected the special neural molecules by immunfluorescence staining. NSCs were cultured on the cell climbing film. After neuronal differentiation, differentiated NSCs were mounted for immunocytochemistry with immunofluorescence technique. RESULTS: The differentiating cells look like neuron, some special factors, such as Glial cell line-derived neurotrophic factor, nerve growth factor, in the media can be detected while differentiated neuron-like cells can express the special neural molecules. CONCLUSION: Differentiation of NSCs towards neurons can be induced by velvet antler polypeptide.

Detection of the Single-Session Complete Ablation Rate by Contrast-Enhanced Ultrasound during Ultrasound-Guided Laser Ablation for Benign Thyroid Nodules: A Prospective Study.

This study aimed to investigate the single-session complete ablation rate of ultrasound-guided percutaneous laser ablation (LA) for benign thyroid nodules. LA was performed in 90 patients with 118 benign thyroid nodules. Contrast-enhanced ultrasound (CEUS) was used to evaluate complete nodule ablation one day after ablation. Thyroid nodule volumes, thyroid functions, clinical symptoms and complications were evaluated 1, 3, 6, 12, and 18 months after ablation. Results showed that all benign thyroid nodules successfully underwent LA. The single-session complete ablation rates for nodules with maximum diameters ≤2 cm, 2-3 cm and ≥3 cm were 93.4%, 70.3% and 61.1%, respectively. All nodule volumes significantly decreased than that one day after ablation (P < 0.05); at the final evaluation, the volume decreased from 6.16 ± 5.21 mL to 0.05 ± 0.01 mL. Thyroid functions did not show significant differences at one month after ablation compared with that before (P > 0.05). Three patients had obvious pain during ablation; one (1.1%) had recurrent laryngeal nerve injury, but the voice returned to normal within 6 months after treatment. Thus, ultrasound-guided LA can effectively inactivate benign thyroid nodules. LA is a potentially viable minimally invasive treatment that offers good cosmetic effects.

4-Carbonyl-2,6-dibenzylidenecyclohexanone derivatives as small molecule inhibitors of STAT3 signaling pathway.

Inhibition of STAT3 signaling pathway is proposed to be a promising strategy for cancer treatment. In this study, a series of 4-carbonyl-2,6-dibenzylidenecyclohexanone derivatives were prepared and evaluated as anticancer agents. The most potent compound 13r was discovered to exhibit antiproliferative activity against a broad rang of cancer cell lines and relatively low cytotoxicity against normal human cells. Besides, 13r effectively suppressed STAT3 expression as well as phosphorylation, and surface plasmon resonance analysis confirmed the direct interaction of 13r with STAT3. Docking simulation showed that 13r could inhibit STAT3 by targeting SH2 domain. This study provided evidence for these compounds to be further developed as antitumor agents through inhibition of the STAT3 pathway.

Diethylstilbestrol Regulates the Expression of LGR8 in Mouse Gubernaculum Testis Cells.

BACKGROUND Hormonal effects on the gubernaculum can affect testicular descent. Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen that disrupts the outgrowth of gubernaculums, leading to testis maldescent. However, the underlying mechanisms remain elusive. MATERIAL AND METHODS The gubernaculum were removed from 3-day-old mice and cultured. The subcultured cells were randomly divided into a normal control group and experimental groups. The DES groups were administered 10 µg/ml, 1 µg/ml, 0.1 µg/ml, 0.01 µg/ml of diethylstilbestrol dissolved in dimethyl sulfoxide (DMSO) respectively. The cell morphology was observed under an inverted microscope, and leucine-rich repeat-containing G protein-coupled receptor 8 (LGR8) was localized by immunofluorescence. The expressions of LGR8 gene and protein in gubernaculum cells were quantified by RT-PCR and Flow Cytometer respectively. RESULTS DES treatment converted cells from a normal fibroblast-like morphology into a more refractile, spindle-shaped morphology or irregular elliptical shapes along with cytoplasmic shrinkage. LGR8 was expressed in the cytoplasmic membrane, DES dose-dependently downregulated LGR8 expression at low doses (≤1.0 µg/ml), but upregulated LGR8 at high doses (10 µg/ml) at both the mRNA and protein levels. CONCLUSIONS These results suggest that DES causes testicular maldescent by altering the LGR8 pathway in mouse gubernaculum testis cells.