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1.
Cancer Res ; 2020 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-32376602

RESUMO

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy characterized by extensive local invasion and systemic spread. In this study, we employed a three-dimensional organoid model of human pancreatic cancer to characterize the molecular alterations critical for invasion. Time lapse microscopy was used to observe invasion in organoids from 25 surgically resected human PDAC samples in collagen I. Subsequent lentiviral modification and small molecule inhibitors were used to investigate the molecular programs underlying invasion in PDAC organoids. When cultured in collagen I, PDAC organoids exhibited two distinct, morphologically defined invasive phenotypes, mesenchymal and collective. Each individual PDAC gave rise to organoids with a predominant phenotype, and PDAC that generated organoids with predominantly mesenchymal invasion showed a worse prognosis. Collective invasion predominated in organoids from cancers with somatic mutations in the driver gene SMAD4 (or its signaling partner TGFBR2). Re-expression of SMAD4 abrogated the collective invasion phenotype in SMAD4-mutant PDAC organoids, indicating that SMAD4 loss is required for collective invasion in PDAC organoids. Surprisingly, invasion in passaged SMAD4-mutant PDAC organoids required exogenous TGFß, suggesting that invasion in SMAD4-mutant organoids is mediated through non-canonical TGFß signaling. The Rho-like GTPases RAC1 and CDC42 acted as potential mediators of TGFß-stimulated invasion in SMAD4-mutant PDAC organoids, as inhibition of these GTPases suppressed collective invasion in our model. These data suggest that PDAC utilizes different invasion programs depending on SMAD4 status, with collective invasion uniquely present in PDAC with SMAD4 loss.

2.
Peptides ; : 170328, 2020 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-32380200

RESUMO

An outbreak caused by 2019 novel coronavirus (2019-nCoV) was first identified in Wuhan City, Hubei Province, China. The new virus was later named SARS-CoV-2. The virus has affected tens of thousands of patients in the world. The infection of SARS-CoV-2 causes severe pneumonia and even death. It is urgently needed to find a therapeutic method to treat patients with SARS-CoV-2 infection. Studies showed that the surface spike (S) protein is essential for the coronavirus binding and entry of host cells. The heptad repeats 1 and 2 (HR1 and HR2) in the S protein play a decisive role in the fusion of the viral membrane with the host cell membrane. We predicted the HR1 and HR2 regions in S protein by sequence alignment. We simulated a computational model of HR1/2 regions and the fusion core. The binding energy of HR1 and HR2 of the fusion core was -33.4kCal/mol. We then designed antivirus peptides by molecular dynamics simulation of the fusion core. The binding energy of HR2-based antiviral peptide to HR1 was -43.0kCal/mol, which was stronger than the natural stage of the fusion core, suggesting that the predicted antiviral peptide can competitively bind with HR1 to prevent forming of the fusion core. The antiviral peptides can prevent SARS-CoV-2 membrane fusion and can potentially be used for the prevention and treatment of infections.

3.
PLoS Biol ; 18(5): e3000746, 2020 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-32453802

RESUMO

Members of the Tre2-Bub2-Cdc16 (TBC) family often function to regulate membrane trafficking and to control signaling transductions pathways. As a member of the TBC family, TBC1D23 is critical for endosome-to-Golgi cargo trafficking by serving as a bridge between Golgi-bound golgin-97/245 and the WASH/FAM21 complex on endosomal vesicles. However, the exact mechanisms by which TBC1D23 regulates cargo transport are poorly understood. Here, we present the crystal structure of the N-terminus of TBC1D23 (D23N), which consists of both the TBC and rhodanese domains. We show that the rhodanese domain is unlikely to be an active sulfurtransferase or phosphatase, despite containing a putative catalytic site. Instead, it packs against the TBC domain and forms part of the platform to interact with golgin-97/245. Using the zebrafish model, we show that impacting golgin-97/245-binding, but not the putative catalytic site, impairs neuronal growth and brain development. Altogether, our studies provide structural and functional insights into an essential protein that is required for organelle-specific trafficking and brain development.

4.
Mol Oncol ; 2020 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-32335998

RESUMO

linc-ROR is reported to be a potential biomarker of breast cancer, but the detailed mechanism of linc-ROR-mediated breast cancer regulation has not been fully studied. We aimed to explore how linc-ROR affects proliferation, metastasis, and drug sensitivity in breast cancer. Cell lines in which linc-ROR was overexpressed or knocked down were constructed, and the cell proliferation, colony formation, cell migration, and invasion abilities of these lines were explored. A CCK-8 assay was performed to determine the sensitivity of the breast cancer cells to rapamycin. Next-generation sequencing was conducted to explore the detailed regulatory mechanism of linc-ROR; differentially expressed RNAs in the linc-ROR-overexpressing cell line compared with the negative control were screened out, and their target genes were chosen to perform Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes analysis, protein-protein interaction network analysis, and competing endogenous RNA (ceRNA) network analysis. The ceRNA mechanism of linc-ROR for miR-194-3p, which targets MECP2, was determined through dual-luciferase reporter assay, RT-qPCR, western blot, and rescue experiments. Finally, we found that linc-ROR was upregulated in breast tumor tissues. linc-ROR promoted the cell proliferation, colony formation, cell migration, and invasion of breast cancer and decreased the sensitivity of breast cancer cells to rapamycin. The overexpression of linc-ROR triggered changes in the whole transcriptome of breast cancer cells, and a total of 85 lncRNAs, 414 microRNAs, 490 mRNAs, and 92 circRNAs were differentially expressed in the linc-ROR-overexpressing cell line compared with the negative control. Through a series of bioinformatic analyses, the 'linc-ROR/miR-194-3p/MECP2' ceRNA regulatory axis was confirmed to be involved in the linc-ROR-mediated progression and drug sensitivity of breast cancer. In conclusion, linc-ROR serves as an onco-lncRNA in breast cancer and promotes the survival of breast cancer cells during rapamycin treatment by functioning as a ceRNA sponge for miR-194-3p, which targets MECP2.

5.
Sci Rep ; 10(1): 3745, 2020 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-32111951

RESUMO

During mung bean post-germination seedling growth, various metabolic and physiological changes occurred, leading to the improvement of its nutritional values. Here, transcriptomic and metabolomic analyses of mung bean samples from 6-hour, 3-day and 6-day after imbibition (6-HAI, 3-DAI, and 6-DAI) were performed to characterize the regulatory mechanism of the primary metabolites during the post-germination seedling growth. From 6-HAI to 3-DAI, rapid changes in transcript level occurred, including starch and sucrose metabolism, glycolysis, citrate cycle, amino acids synthesis, and plant hormones regulation. Later changes in the metabolites, including carbohydrates and amino acids, appeared to be driven by increases in transcript levels. During this process, most amino acids and monosaccharides kept increasing, and accumulated in 6-day germinated sprouts. These processes were also accompanied with changes in hormones including abscisic acid, gibberellin, jasmonic acid, indole-3-acetic acid, etc. Overall, these results will provide insights into molecular mechanisms underlying the primary metabolic regulation in mung bean during post-germination seedling growth.

6.
Med Sci Monit ; 26: e922925, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-32134048

RESUMO

BACKGROUND Anatomical reconstruction using a semitendinosus tendon autograft is one of the most widely-used techniques for chronic lateral ankle instability (CLAI), and it can result in good biomechanical recovery for patients. The purpose of this study was to investigate the outcome of a novel individualized three-dimensional printed guide template for lateral ankle ligament reconstruction compared with the traditional surgical methods. MATERIAL AND METHODS We retrospectively studied 34 patients with CLAI who required lateral ankle ligament reconstruction. Patients were randomly divided into 2 cohorts: the template group (18 patients) and the conventional group (16 patients). The average operation duration and number of radiation exposures were compared between the 2 cohorts. The displacement of anterior talar and talar tilt angle were recorded at the last follow-up, and Karlsson-Peterson score and American Orthopedic Foot and Ankle Society Score (AOFAS) were also compared. RESULTS All patients had satisfactory ankle stability at the last follow-up. The average operation duration was 51.9±3.6 min and the average number of radiation exposures was 1.34±0.6 in the template group, and the average operation duration was 72.4±12.6 min and the average number of radiation exposures was 6.58±1.7 in the conventional group. Difference between the 2 cohorts was statistically significant. However, in AOFAS (95.2±2.5 vs. 94.9±2.2; P>0.01.) and Karlsson Score (94.7±3.6 vs. 93.8±4.1; P>0.01.), no significant differences were found between the 2 cohorts. CONCLUSIONS Both the template technique and the conventional method provided satisfactory outcomes for CLAI patients. However, the shorter operation duration and low number of radiation exposures in the template cohort suggest it is the better alternative for treatment of CLAI.


Assuntos
Instabilidade Articular/cirurgia , Ligamentos Laterais do Tornozelo/cirurgia , Impressão Tridimensional , Tendões/transplante , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Procedimentos Cirúrgicos Reconstrutivos , Estudos Retrospectivos , Cirurgia Assistida por Computador , Transplante Autólogo , Adulto Jovem
7.
PLoS One ; 15(2): e0229205, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32092096

RESUMO

BACKGROUND: Given their geographical proximity but differences in cultural and religious dietary customs, we hypothesize that children from the three main ethnic populations (Han, Hui, and Tibetan) residing in the Qinghai-Tibetan Plateau region differs in their non-iatrogenic antibiotic loads. METHODS: To determine the antibiotic burden of the school children unrelated to medical treatment, we quantified the antibiotic residues in morning urine samples from 92 Han, 72 Tibetan, and 85 Muslim Hui primary school children aged 8 to 12 years using high-performance liquid chromatography-tandem mass spectrometry, and performed correlation analysis between these data and concurrent dietary nutrition assessments. RESULTS: Sixteen of the 18 targeted antibiotics (4 macrolides, 3 ß-lactams, 2 tetracyclines, 4 quinolones, 3 sulfonamides, and 2 aminoanols) were identified in the urine samples with an overall detection frequency of 58.63%. The detection frequency of the six antibiotic classes ranged from 1.61% to 32.53% with ofloxacin showing the single highest frequency (18.47%). Paired comparison analysis revealed significant differences in antibiotic distribution frequency among groups, with Tibetans having higher enrofloxacin (P = 0.015) and oxytetracycline (P = 0.021) than Han children. Norfloxacin (a human/veterinary antibiotic) was significantly higher in the Hui children than in the Han children (P = 0.024). Dietary nutrient intake assessments were comparable among participants, showing adequate levels of essential vitamins and minerals across all three ethnic groups. However, significant differences in specific foods were observed among groups, notably in lower fat consumption in the Hui group. CONCLUSIONS: The introduction and accumulation of antibiotic residues in school children through non-iatrogenic routes (food or environmental sources) poses a serious potential health risk and merits closer scrutiny to determine the sources. While the exact sources of misused or overused antibiotics remains unclear, further study can potentially correlate ethnicity-specific dietary practices with the sources of contamination.

8.
Cancer Med ; 9(5): 1683-1693, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31945265

RESUMO

BACKGROUND: Pembrolizumab (Pembro) in combination with chemotherapy has been approved for the treatment of pretreated advanced NSCLC in the United States and China for its significant efficacy. However, the cost-effectiveness is unknown considering Pembro's high price. The impact of programmed death ligand 1 (PD-L1) test on the cost-effectiveness is also unknown. The current study assessed the cost-effectiveness of combination therapy for nonsquamous NSCLC from the United States and China public payers' perspective. MATERIALS AND METHODS: A literature-based Markov model was conducted using KEYNOTE-189 trial data to compare cost and quality-adjusted life years (QALYs) of three treatment strategies for nonsquamous NSCLC: Pembro-chemotherapy combination and chemotherapy strategy without PD-L1 test, and treatment strategy according to their PD-L1 status. RESULTS: In base case analysis, the combination strategy generated an additional 0.78 QALYs and 0.59 QALYs over chemotherapy in the United States and China respectively, resulting in an ICER of $132 392/QALY in the United States and $92 533/QALY in China. In the PD-L1 ≥1% base case, the ICERs were $77 754/QALY and $56 768/QALY respectively in the United States and China for PD-L1 test strategy. In the PD-L1 ≥50% base case, the ICERs were $44 731/QALY and $34 388/QALY respectively in the United States and China for PD-L1 test strategy. Lowering Pembro price can also partly decrease the ICERs. CONCLUSION: Compared with chemotherapy, the combination strategy is not cost-effective for the treatment of NSCLC in the American and Chinese health care system at WTP threshold of $100 000/QALY for the United States and $27 351/QALY for China. Using PD-L1 test for patient selection and price reduction could improve the cost-effective probabilities of immunotherapy for nonsquamous NSCLC.

9.
Andrology ; 8(1): 110-116, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31127676

RESUMO

BACKGROUND: It has been reported that paternal folic acid deficiency is correlated with male infertility and increased birth defects in the offspring. However, there are few data concerning the influence of folic acid supplementation on male-factor infertility with MTHFR gene polymorphisms. OBJECTIVES: To evaluate whether folic acid supplementation has a beneficial effect on oligozoospermia with MTHFR gene polymorphisms in Chinese infertility population. MATERIALS AND METHODS: The infertile men suffering oligozoospermia with MTHFR gene polymorphisms were randomly divided into the folic acid treatment groups receiving folic acid 0.8 mg daily for 3 months and the placebo groups receiving placebo for 3 months. Semen parameters, seminal MDA, and DNA fragmentation were measured. Furthermore, spontaneous pregnancy rate and live birth rate were evaluated. RESULTS: Administration of folic acid for 3 months could significantly improve the seminal parameters in patients with MTHFR 677 TT genotype in comparison with that receiving placebo. Moreover, seminal MDA and sperm DNA fragmentation index in patients with MTHFR 677 TT genotype significantly declined at the end of treatment. Spontaneous pregnancy rate and live birth rate tended to be significantly higher in couples in which the men with MTHFR 677 TT genotype receiving folic acid than that receiving placebo. However, folic acid treatment did not exhibit any advantage in MTHFR 677 CT, 1298 AC, 1298 CC, 1793 GA, or combined 677 CT/1298 AC genotype. DISCUSSION: The anti-oxidation function of folic acid is one of possible mechanisms invovled in improving seminal parameters and pregnancy outcome. CONCLUSIONS: Folic acid supplementation has a beneficial effect on oligozoospermia with MTHFR 677 TT genotype in term of seminal parameters, seminal MDA, sperm DNA fragmentation, and pregnancy outcome.

10.
Arch Biochem Biophys ; 680: 108225, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31838119

RESUMO

An increase in intracellular Cl- concentration ([Cl-]i) may be a general response of airway epithelial cells to various stimuli and may participate in some basic cellular functions. However, whether the basic functional activities of cells, such as proliferation and wound healing, are related to Cl- activities remains unclear. This study aimed to investigate the effects and potential mechanisms of [Cl-]i on the proliferation and wound healing ability of airway epithelial BEAS-2B cells. BEAS-2B cells were treated with four Cl- channel inhibitors (T16Ainh-A01, CFTRinh-172, CaCCinh-A01, and IAA-94), and the Cl- fluorescence probe N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide was used. Results showed that all Cl- channel inhibitors could increase [Cl-]i in BEAS-2B cells. The increased [Cl-]i induced by Cl- channel inhibitors or clamping [Cl-]i at high levels enhanced the phosphorylation of focal adhesion kinase (FAK) and subsequently promoted the proliferation and wound healing ability of BEAS-2B cells. By contrast, the FAK inhibitor PF573228 abrogated these effects induced by the increased [Cl-]i. FAK also activated the PI3K/AKT signaling pathway. In conclusion, increased [Cl-]i promotes the proliferation and wound healing ability of BEAS-2B cells by activating FAK to activate the PI3K/AKT signaling pathway. Intracellular Cl- may act as a signaling molecule to regulate the proliferation and wound healing ability of airway epithelial cells.

11.
Angew Chem Int Ed Engl ; 59(9): 3544-3548, 2020 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-31880061

RESUMO

Heterostructured Mo2 C-MoOx on carbon cloth (Mo2 C-MoOx /CC), as a model of easily oxidized electrocatalysts under ambient conditions, is investigated to uncover surface reconfiguration during the hydrogen evolution reaction (HER). Raman spectroscopy combined with electrochemical tests demonstrates that the MoVI oxides on the surface are in situ reduced to MoIV , accomplishing promoted HER in acidic condition. As indicated by density functional theoretical calculations, the in situ reduced surface with terminal Mo=O moieties can effectively bring the negative ΔGH* on bare Mo2 C close to a thermodynamic neutral value, addressing difficult H* desorption toward fast HER kinetics. The optimized Mo2 C-MoOx /CC only requires a low overpotential (η10 ) of 60 mV at -10 mA cm-2 in 1.0 m HClO4 , outperforming Mo2 C/CC and most non-precious electrocatalysts. In situ surface reconfiguration are shown on W2 C-WOx , highlighting the significance to boost various metal-carbides and to identify active sites.

12.
Molecules ; 24(24)2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31835666

RESUMO

Foliage of jujube (Ziziphus jujuba Mill.) as a byproduct of agriculture, is a traditional nutraceutical material in China. Previous studies have shown that it is a rich resource of polyphenols. However, information on its complete phenolic profile and the difference between cultivars is still limited. This study investigated and compared phytochemical profiles of leaves of 66 Chinese jujube cultivars. Forty-two compounds, including 22 flavonols, two flavanols, one flavanone, 13 derivatives of phenolic acids, three simple acids, and one unknown hexoside were identified/tentatively identified using high-performance liquid chromatography (HPLC) coupled with high-resolution mass spectrometry. Eight major flavonols were quantified by HPLC coupled with an ultraviolet (UV) detector. The contents of total flavonoids ranged from 2.6-25.1 mg/g dry weight (DW). Differences between cultivars were analyzed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). This study presents a systematic study on the phenolic compounds in Chinese jujube leaves of different cultivars.

13.
Mol Ther Nucleic Acids ; 19: 228-239, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31846800

RESUMO

Treatment of pancreatic ductal adenocarcinoma (PDAC) remains a clinical challenge. There is an urgent need to develop novel strategies to enhance survival and improve patient prognosis. MicroRNAs (miRNAs) play critical roles as oncogenes or tumor suppressors in the regulation of cancer development and progression. In this study, we demonstrate that low expression of miR-15a is associated with poor prognosis of PDAC patients. miR-15a expression is reduced in PDAC while closely related miR-16 expression remains relatively unchanged. miR-15a suppresses several important targets such as Wee1, Chk1, Yap-1, and BMI-1, causing cell cycle arrest and inhibiting cell proliferation. Ectopic expression of miR-15a sensitizes PDAC cells to gemcitabine reducing the half maximal inhibitory concentration (IC50) more than 6.5-fold. To investigate the therapeutic potential of miR-15a, we used a modified miR-15a (5-FU-miR-15a) with uracil (U) residues in the guide strand replaced with 5-fluorouracil (5-FU). We demonstrated enhanced inhibition of PDAC cell proliferation by 5-FU-miR-15a compared to native miR-15a. In vivo we showed the therapeutic power of 5-FU-miR-15a alone or in combination with gemcitabine with near complete elimination of PDAC lung metastatic tumor growth. These results support the future development of 5-FU-miR-15a as a novel therapeutic agent as well as a prognostic biomarker in the clinical management of PDAC.

14.
Medicine (Baltimore) ; 98(47): e17875, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31764782

RESUMO

BACKGROUND: This study aimed to explore the effect of YHJD (Yiqi Huayu Jiedu decoction) in patients with stages II and III gastric cancer. METHODS: A multicenter, prospective, cohort study was conducted in Jiangsu Province Hospital of Traditional Chinese Medicine, Jiangsu Cancer Hospital, Nanjing Drum Tower Hospital, People's Liberation Army Bayi Hospital, Changzhou Traditional Chinese Medicine Hospital, Changzhou Tumor Hospital, Traditional Chinese Medicine Hospital of Kunshan, Yangzhou Hospital of Traditional Chinese Medicine, and Yixing Tumor Hospital. A total of 489 patients with stage II or III gastric cancer were enrolled after radical gastrectomy. Among them, 238 were included in the chemotherapy group (received chemotherapy alone) and 251 in the YHJD group (received chemotherapy combined with YHJD). The DFS (disease-free survival) rate, 5-year survival rate, quality of life, and traditional Chinese medicine (TCM) symptoms of the 2 groups were compared. RESULTS: The DFS curve of the YHJD group was higher than that of the chemotherapy group (P = .0042). The HR (hazard ratio) was 0.672, and its corresponding 95% CI (confidence interval) was 0.511 to 0.884. For stage II patients, the P value was .8323, which indicated that the difference was not significant. The risk HR was 0.938, and the corresponding 95% CI was 0.521 to 1.689. For stage III patients, the P value was .0072, indicating a statistically significant difference. The HR was 0.653, and the corresponding 95% CI was 0.477 to 0.893. The 5-year survival rate of the YHJD group was 85.29%, which was higher than that of the chemotherapy group (71.05%). Compared with the chemotherapy group, the YHJD group had better quality of life and lower TCM symptom scores. CONCLUSION: YHJD decoction is effective in improving DFS rate in patients with gastric cancer stage III after radical gastrectomy. Moreover, it can reduce the risk of recurrence and metastasis and improve the quality of life in patients with gastric cancer stage II or III after radical gastrectomy.


Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Neoplasias Gástricas/tratamento farmacológico , Adulto , Idoso , Estudos de Coortes , Terapia Combinada , Feminino , Gastrectomia , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Estudos Prospectivos , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Resultado do Tratamento , Adulto Jovem
15.
Biomed Pharmacother ; 120: 109507, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31590125

RESUMO

LINC01234 plays a pivot role in the tumorigenesis of gastric cancer, colon cancer and lung cancer. However, how LINC01234 participates in oral squamous cell carcinoma (OSCC) progression remains unknown. In our research, we showed that LINC01234 was dramatically upregulated in OSCC tissues. And interestingly, high LINC01234 expression predicted a low overall survival rate in OSCC patients. Knockdown of OSCC inhibited the proliferation of cancer cells and led to more cells restricted in G0 phase. Moreover, LINC01234 silencing decreased the migration and invasion of OSCC cells. Additionally, downregulation of LINC01234 limited OSCC tumor propagation in vivo. Mechanistic investigation elucidated that LINC01234 inhibited the activity of miR-637 to increase the expression of NUPR1. Via upregulating NUPR1 level, LINC01234 contributed to malignant behaviors of OSCC cells. Collectively, our research shows that LINC01234 exerts an important role in OSCC progression via miR-637/NUPR1 axis.

16.
Proc Natl Acad Sci U S A ; 116(45): 22598-22608, 2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31624125

RESUMO

Pontocerebellar hypoplasia (PCH) is a group of neurological disorders that affect the development of the brain, in particular, the pons and cerebellum. Homozygous mutations of TBC1D23 have been found recently to lead to PCH; however, the underlying molecular mechanisms remain unclear. Here, we show that the crystal structure of the TBC1D23 C-terminal domain adopts a Pleckstrin homology domain fold and selectively binds to phosphoinositides, in particular, PtdIns(4)P, through one surface while binding FAM21 via the opposite surface. Mutation of key residues of TBC1D23 or FAM21 selectively disrupts the endosomal vesicular trafficking toward the Trans-Golgi Network. Finally, using the zebrafish model, we show that PCH patient-derived mutants, impacting either phosphoinositide binding or FAM21 binding, lead to abnormal neuronal growth and brain development. Taken together, our data provide a molecular basis for the interaction between TBC1D23 and FAM21, and suggest a plausible role for PtdIns(4)P in the TBC1D23-mediating endosome-to-TGN trafficking pathway. Defects in this trafficking pathway are, at least partially, responsible for the pathogenesis of certain types of PCH.

17.
Artigo em Inglês | MEDLINE | ID: mdl-31636680

RESUMO

HongJing I (HJI), a traditional Chinese herbal formula, has been confirmed to be effective for the clinical treatment of erectile dysfunction (ED). However, the mechanism of action of HJI remains unclear. Here, we aimed to investigate the effect and underlying mechanisms of HJI against ED in a rat model of bilateral cavernous nerve injury (BCNI). Rats were divided into five groups: normal control (NC), BCNI-induced ED model (M), M + low-dose HJI (HL), M + medium-dose HJI (HM), and M + high-dose HJI (HH). All groups were treated with normal saline or the relevant drug for 28 consecutive days after inducing BCNI-ED. At the end of the treatment period, the intracavernous pressure (ICP) was recorded, and histological examination was conducted using Masson's trichrome staining. Immunofluorescence staining and western blotting were applied to detect the changes in fibrosis protein and Ras homolog A (RhoA), Rho-associated protein kinase 1 (ROCK1), and ROCK2 expression. We found that HJI effectively improved the ICP in the treatment groups. In addition, RhoA, ROCK1, and ROCK2 expression levels were increased upon BCNI-ED induction, and HJI successfully inhibited cavernosum fibrosis and the activation of RhoA/ROCK2 signaling. Overall, these results suggest that the effects of HJI in attenuating ED may be caused, at least in part, by the suppression of RhoA/ROCK2 signaling and alleviation of fibrosis. However, the precise mechanism surrounding this requires further investigation in future studies.

18.
Mol Med Rep ; 20(6): 5050-5058, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31638214

RESUMO

Ghrelin is an orexigenic hormone that is produced by gastric cells. Ghrelin stimulates food intake and increases gastric movement. In rat model, injected ß­hydroxybutyric acid (ß­HB) leads to a decrease in body weight. It has been reported that patients with gastric erosions are slower to evacuate the stomach. The aim of the present study was to investigate the effects of ghrelin and ß­HB on motility and inflammation in rat gastric antral smooth muscle cells (GASMCs). GASMCs were extracted from rat gastric antrum. Cell viability was determined using the Cell Counting Kit­8 assay. A reactive oxygen species (ROS) assay kit was used to analyze the levels of ROS using flow cytometry. Protein levels were determined using western blotting, and the expression levels of mRNAs were evaluated using reverse transcription­quantitative PCR. ß­HB inhibited the expression of myosin regulatory light polypeptide 9 (MYL9), myosin light chain kinase (MLCK), transforming protein RhoA (RhoA), Rho­associated protein kinase­1 (ROCK­1) and growth hormone secretagogue receptor (GHS­R). By contrast, ghrelin increased the expression of MYL9, MLCK, RhoA, ROCK­1 and GHS­R in ß­HB­treated GASMCs. ß­HB increased the levels of tumor necrosis factor (TNF)­α, interleukin (IL)­6 and ROS, and decreased the levels of manganese (Mn) superoxide dismutase (SOD), copper/zinc (Cu/Zn)SOD and catalase. Ghrelin decreased the expression of TNF­α, IL­6, ROS and catalase, whereas ghrelin promoted the expression of MnSOD and Cu/ZnSOD in ß­HB­treated GASMCs. Short interfering RNA targeting GHS­R inhibited the expression of MYL9, MLCK, RhoA and ROCK­1, and increased the levels of TNF­α, IL­6 and ROS in ß­HB­treated or ghrelin­treated GASMCs. The present study provided preliminary evidence that ß­HB inhibits the motility of GASMCs and promotes inflammation in GASMCs, whereas ghrelin decreases these effects. GHS­R acted as a primary regulator of motility and inflammation in GASMCs treated with ß­HB and ghrelin.

19.
Sex Med ; 7(4): 433-440, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31540881

RESUMO

INTRODUCTION: Cavernosal nerve (CN) injury is commonly caused by radical prostatectomy surgery, and it might directly lead to erectile dysfunction (ED). Currently, the role of mitogen-activated protein kinase (MAPK) family proteins in phenotypic transformation of corpus cavernosum smooth muscle cell (CCSMC) after CNs injury is poorly understood. AIM: To investigate the role of p38 MAPK in hypoxia-induced phenotypic transformation of CCSMCs after CN injury. METHODS: In total, 20 Sprague-Dawley rats (male and 8 weeks of age) were randomly divided into 2 groups, including a sham group and CNCI group. In the sham group, rats were sham-operated by identifying 2 CNs without causing direct damage to the CNs. In the CNCI group, rats were subjected to bilateral CN crush injury. CCSMCs were isolated from the normal corpus cavernosum tissues of the Sprague-Dawley rat and then cultured in 21% or 1% O2 concentration context for 48 hours. MAIN OUTCOME MEASURES: Intracavernous pressure/mean arterial pressure were analyzed to measure erectile response. The impact of hypoxia on penile pathology, as well as the expression of extracellular signal-regulated kinases, the c-Jun NH2-terminal kinase, and p38 MAPK, were analyzed. RESULTS: Compared with the sham group, the intracavernous pressure/mean arterial pressure rate and α-smooth muscle actin expression of CNCI group were decreased significantly (P = .0001; P = .016, respectively), but vimentin expression was significantly increased (P = .023). Phosphorylated p38 level in CNCI group was decreased significantly (P = .017; sham: 0.17 ± 0.005; CNCI: 0.14 ± 0.02). The CCSMCs in the normoxia group were long fusiform, whereas the morphology of CCSMCs in the hypoxia group became hypertrophic. After hypoxia for 48 hours, the expression of α-smooth muscle actin and phosphorylated p38 MAPK was decreased significantly (P = .01; P = .024, normoxia: 0.66 ± 0.18, hypoxia: 0.26 ± 0.08, respectively), and the expression of hypoxia-inducible factor-1α and collagen I was increased significantly in hypoxia group (P = .04; P = .012, respectively). CONCLUSIONS: Hypoxia induced the phenotypic transformation of CCSMCs after CNCI might be associated with the downregulation of phosphorylated p38 MAPK. Chen S, Huang X, Kong X, et al. Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating p38 Mitogen-Activated Protein Kinase Expression. Sex Med 2019;7:433-440.

20.
Front Pharmacol ; 10: 673, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31258479

RESUMO

Background: Programmed cell death 1 (PD-1) or programmed cell death ligand 1 (PD-L1) inhibitions are being strongly recommended for the treatment of various cancers, while the efficacy of PD-1/PD-L1 inhibitions varies from individuals. It is urgent to explore some biomarkers to screen the most appropriate cancer patients. Tumor mutation burden (TMB) as a potential alternative has been drawing more and more attention. Therefore, we conducted a meta-analysis to quantitatively explore the association between TMB and outcomes of PD-1/PD-L1 inhibitions. Methods: We searched eligible studies that evaluated the association between TMB and the outcomes of PD-1/PD-L1 inhibitions from PubMed, Embase, and Cochrane database up to October 2018. The primary endpoints were the progression-free survival (PFS) and the overall survival (OS) in patients with high TMB or low TMB. The pooled hazard ratios (HR) for PFS and OS were performed by Stata. Results: In this analysis, a total of 2,661 patients from eight studies were included. Comparing PD-1/PD-L1 inhibitions to chemotherapy, the pooled HR for PFS and OS in patients with high TMB was 0.66 [95% confidence interval (CI) 0.50 to 0.88; P = 0.004] and 0.73 (95% CI 0.50 to 1.08; P = 0.114), respectively, while the pooled HR for PFS and OS in patients with low TMB was 1.38 (95% CI 0.82 to 2.31; P = 0.229) and 1.00 (95% CI 0.80 to 1.24; P = 0.970), respectively. Meanwhile, comparing patients with high TMB to patients with low TMB, the pooled HR for PFS in patients treated with PD-1/PD-L1 inhibitions was 0.47 (95% CI 0.35 to 0.63; P = 0.000). Patients with high TMB showed significant benefits from PD-1/PD-L1 inhibitions compared to patients with low TMB. Conclusion: Despite the present technical and practical barriers, TMB may be a preferable biomarker to optimize the efficacy of PD-1/PD-L1 inhibitions.

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