ABSTRACT
INTRODUCTION: Airway epithelial cells are recognised as an essential controller for the initiation and perpetuation of asthmatic inflammation, yet the detailed mechanisms remain largely unknown. This study aims to investigate the roles and mechanisms of the mechanistic target of rapamycin (MTOR)-autophagy axis in airway epithelial injury in asthma. METHODS: We examined the MTOR-autophagy signalling in airway epithelium from asthmatic patients or allergic mice induced by ovalbumin or house dust mites, or in human bronchial epithelial (HBE) cells. Furthermore, mice with specific MTOR knockdown in airway epithelium and autophagy-related lc3b-/- mice were used for allergic models. RESULTS: MTOR activity was decreased, while autophagy was elevated, in airway epithelium from asthmatic patients or allergic mice, or in HBE cells treated with IL33 or IL13. These changes were associated with upstream tuberous sclerosis protein 2 signalling. Specific MTOR knockdown in mouse bronchial epithelium augmented, while LC3B deletion diminished allergen-induced airway inflammation and mucus hyperproduction. The worsened inflammation caused by MTOR deficiency was also ameliorated in lc3b-/- mice. Mechanistically, autophagy was induced later than the emergence of allergen-initiated inflammation, particularly IL33 expression. MTOR deficiency increased, while knocking out of LC3B abolished the production of IL25 and the eventual airway inflammation on allergen challenge. Blocking IL25 markedly attenuated the exacerbated airway inflammation in MTOR-deficiency mice. CONCLUSION: Collectively, these results demonstrate that allergen-initiated inflammation suppresses MTOR and induces autophagy in airway epithelial cells, which results in the production of certain proallergic cytokines such as IL25, further promoting the type 2 response and eventually perpetuating airway inflammation in asthma.
Subject(s)
Asthma/metabolism , Inflammation/metabolism , Interleukin-17/biosynthesis , Interleukins/metabolism , TOR Serine-Threonine Kinases/metabolism , Adult , Aged , Allergens , Animals , Asthma/pathology , Asthma/physiopathology , Autophagy/drug effects , Cells, Cultured , Epithelial Cells/metabolism , Female , Gene Knockdown Techniques , Humans , Inflammation/pathology , Interleukin-13/metabolism , Interleukin-13/pharmacology , Interleukin-33/metabolism , Interleukin-33/pharmacology , Male , Mice , Microtubule-Associated Proteins/genetics , Middle Aged , Respiratory Mucosa/physiopathology , Signal Transduction , TOR Serine-Threonine Kinases/genetics , Tuberous Sclerosis Complex 2 Protein/metabolismABSTRACT
BACKGROUND: Mothers' knowledge of neonatal jaundice (NNJ) is grossly deficient or inaccurate, which may adversely affect the actions of mothers in the recognition of NNJ and cause a delay in seeking medical attention. MATERIAL AND METHODS: A total of 1036 primiparas were separated randomly into the intervention group and the control group, with 518 primiparas in each group. RESULTS: All (100%) mothers in the intervention group understood that NNJ is a yellow discoloration of the skin and sclera; 94.19% of them considered that NNJ is a common problem in newborns; 82.80% and 95.27% replied that jaundice appearing within the first 36 hours and lasting more than 2 weeks usually indicates pathological NNJ; 96.34%, 80.86%, and 90.32% realized that premature newborns, low birth weight, and perinatal asphyxia, respectively, are more likely to be accompanied by NNJ; 97.41%, 78.71%, and 64.95% knew that maternal-fetal blood group incompatibility, infection, and glucose-6-phosphate dehydrogenase deficiency, respectively, are the common inducements to NNJ; 94.84% could associate NNJ with brain damage; 92.26%, 93.12%, and 74.62% agreed that phototherapy, strengthen feeding, and exchange blood transfusion, respectively, can greatly relieve NNJ. However, some respondents in the control group responded in other ways, such as stopping breastfeeding (9.19%), placing newborns in sunlight (10.24%) and traditional Chinese medicine (10.24%), which was significantly higher than that of the intervention group. There was also a significant delay for respondents in the control group in consulting a pediatrician, and 6.30% of them did not seek medical help until after the interview. CONCLUSIONS: Prenatal training could significantly improve new mothers' understanding of NNJ.
Subject(s)
Health Knowledge, Attitudes, Practice , Jaundice, Neonatal/psychology , Mothers/psychology , Prenatal Education/statistics & numerical data , Adult , China , Demography , Female , Humans , Parity , Pregnancy , Socioeconomic FactorsABSTRACT
OBJECTIVE: To investigate the feasibility and advantages of transurethral transumbilical laparoendoscopic single-site surgery (TU-LESS) for radical prostatectomy. METHODS: Five patients with prostate cancer underwent TU-LESS for radical prostatectomy, with a four-channel single-port device inserted into a 2. 5 cm periumbilical incision and another placed through the urethra, followed by analysis of the perioperative data. RESULTS: All the operations were successfully accomplished, with neither conversion to open surgery nor additional channel. The mean operation time, intraoperative blood loss, and postoperative hospital stay were 168 min, 120 ml, and 15 d, respectively. No severe perioperative complications were observed. TNM stage classification manifested T2cN0M0 in 2 cases and T2bN0M0 in the other 3. Postoperative pathology showed no negative surgical margins in any of the cases. CONCLUSION: TU-LESS is safe and feasible for radical prostatectomy and can reduce the complication of low urinary tract surgery by single-site laparoendoscopy.
Subject(s)
Laparoscopy , Prostatectomy/methods , Prostatic Neoplasms/surgery , Blood Loss, Surgical , Feasibility Studies , Humans , Length of Stay , Male , Natural Orifice Endoscopic Surgery/methods , Operative Time , Umbilicus/surgery , Urologic Surgical Procedures, Male/methodsABSTRACT
BACKGROUND: Papillary thyroid carcinoma (PTC) is the most common malignant endocrine tumour, and its incidence and prevalence are increasing considerably. Cellular heterogeneity in the tumour microenvironment is important for PTC prognosis. Spatial transcriptomics is a powerful technique for cellular heterogeneity study. METHODS: In conjunction with a clinical pathologist identification method, spatial transcriptomics was employed to characterise the spatial location and RNA profiles of PTC-associated cells within the tissue sections. The spatial RNA-clinical signature genes for each cell type were extracted and applied to outlining the distribution regions of specific cells on the entire section. The cellular heterogeneity of each cell type was further revealed by ContourPlot analysis, monocle analysis, trajectory analysis, ligand-receptor analysis and Gene Ontology enrichment analysis. RESULTS: The spatial distribution region of tumour cells, typical and atypical follicular cells (FCs and AFCs) and immune cells were accurately and comprehensively identified in all five PTC tissue sections. AFCs were identified as a transitional state between FCs and tumour cells, exhibiting a higher resemblance to the latter. Three tumour foci were shared among all patients out of the 13 observed. Notably, tumour foci No. 2 displayed elevated expression levels of genes associated with lower relapse-free survival in PTC patients. We discovered key ligand-receptor interactions, including LAMB3-ITGA2, FN1-ITGA3 and FN1-SDC4, involved in the transition of PTC cells from FCs to AFCs and eventually to tumour cells. High expression of these patterns correlated with reduced relapse-free survival. In the tumour immune microenvironment, reduced interaction between myeloid-derived TGFB1 and TGFBR1 in tumour focus No. 2 contributed to tumourigenesis and increased heterogeneity. The spatial RNA-clinical analysis method developed here revealed prognosis-associated cellular heterogeneity in the PTC microenvironment. CONCLUSIONS: The occurrence of tumour foci No. 2 and three enhanced ligand-receptor interactions in the AFC area/tumour foci reduced the relapse-free survival of PTC patients, potentially leading to improved prognostic strategies and targeted therapies for PTC patients.
Subject(s)
Thyroid Neoplasms , Humans , Thyroid Cancer, Papillary/genetics , Thyroid Cancer, Papillary/metabolism , Thyroid Cancer, Papillary/pathology , Thyroid Neoplasms/diagnosis , Thyroid Neoplasms/genetics , Thyroid Neoplasms/metabolism , Ligands , Tumor Microenvironment/genetics , Neoplasm Recurrence, Local , Gene Expression Profiling , Prognosis , RNAABSTRACT
OBJECTIVE: To explore the expressions of indoleamine 2, 3-dioxygenase (IDO) in hepatocellular carcinoma and analyze its relationship with clinicopathological parameters. METHODS: Quantitative real-time polymerase chain reaction (PCR), fluorogenic quantitative PCR, immunohistochemical and immunofluorescence were used to detect the expression of indoleamine 2, 3-dioxygenase in hepatocellular carcinoma. RESULTS: The IDO mRNA expression in cancerous tissues increased markedly than that in the corresponding non-cancerous tissues (2(-ΔΔCT) = 1.71, P = 0.001) . The immunohistochemical and immunofluorescence results showed that IDO protein was expressed in cytoplasm of hepatocellular carcinoma and tumor-surrounding tissues. But there was no expression in normal liver tissues from benign hepatic lesions and corresponding non-cancerous tissues. An over-expression of IDO protein was detected in 43 patients (48.3%) , a low expression in 25 (28.1%) and no expression in 21 (23.6%). Relationship between IDO expression and clinicopathological parameters: an over-expression of IDO in HCC was associated with recurrence, survival time, metastasis and TNM stage (P < 0.05), but not associated with patient's cirrhosis, AFP level, histological differentiation type, Barcelona clinic liver cancer stage, gender, age, HbsAg positivity, number of tumors and tumor size (P > 0.05). CONCLUSION: An over-expression of IDO in HCC patients may affect patient prognosis.
Subject(s)
Carcinoma, Hepatocellular/enzymology , Indoleamine-Pyrrole 2,3,-Dioxygenase/metabolism , Liver Neoplasms/enzymology , Humans , Liver Neoplasms/pathology , Neoplasm Recurrence, Local , Neoplasm Staging , Prognosis , Real-Time Polymerase Chain ReactionABSTRACT
Introduction: Cochlear implantation is currently the most successful intervention for severe-to-profound sensorineural hearing loss, particularly in deaf infants and children. Nonetheless, there remains a significant degree of variability in the outcomes of CI post-implantation. The purpose of this study was to understand the cortical correlates of the variability in speech outcomes with a cochlear implant in pre-lingually deaf children using functional near-infrared spectroscopy (fNIRS), an emerging brain-imaging technique. Methods: In this experiment, cortical activities when processing visual speech and two levels of auditory speech, including auditory speech in quiet and in noise with signal-to-noise ratios of 10 dB, were examined in 38 CI recipients with pre-lingual deafness and 36 normally hearing children whose age and sex matched CI users. The HOPE corpus (a corpus of Mandarin sentences) was used to generate speech stimuli. The regions of interest (ROIs) for the fNIRS measurements were fronto-temporal-parietal networks involved in language processing, including bilateral superior temporal gyrus, left inferior frontal gyrus, and bilateral inferior parietal lobes. Results: The fNIRS results confirmed and extended findings previously reported in the neuroimaging literature. Firstly, cortical responses of superior temporal gyrus to both auditory and visual speech in CI users were directly correlated to auditory speech perception scores, with the strongest positive association between the levels of cross-modal reorganization and CI outcome. Secondly, compared to NH controls, CI users, particularly those with good speech perception, showed larger cortical activation in the left inferior frontal gyrus in response to all speech stimuli used in the experiment. Discussion: In conclusion, cross-modal activation to visual speech in the auditory cortex of pre-lingually deaf CI children may be at least one of the neural bases of highly variable CI performance due to its beneficial effects for speech understanding, thus supporting the prediction and assessment of CI outcomes in clinic. Additionally, cortical activation of the left inferior frontal gyrus may be a cortical marker for effortful listening.
Subject(s)
Influenza A virus/genetics , Influenza in Birds/virology , Poultry/virology , Animals , China , Humans , Influenza A virus/pathogenicity , Influenza in Birds/epidemiology , Influenza in Birds/genetics , Influenza in Birds/pathology , Influenza, Human/epidemiology , Influenza, Human/genetics , Influenza, Human/pathology , Zoonoses/genetics , Zoonoses/pathologyABSTRACT
OBJECTIVE: To evaluate whether the clinical characteristics of chronic cough were helpful in determining its specific causes. METHODS: Patients with chronic cough were evaluated by a validated systematic diagnostic protocol. The patients with identified single cause were divided into 4 groups accordingly: cough-variant asthma (CVA), upper airway cough syndrome (UACS) or post-nasal drip syndrome (PNDS), eosinophilic bronchitis (EB), gastroesophageal reflux related cough (GERC), and the characteristics of the timing, character, onset and associated manifestations of chronic cough in different causes were compared. RESULTS: A total of 196 patients met the inclusion criteria, including 55 with EB, 45 with UACS, 50 with CVA and 46 with GERC. No significant difference was found in age, gender and course among EB, UACS, CVA and GERC. The incidence of nocturnal cough in CVA was 26.0% (13/44), significantly higher than in EB (9.1% (5/55), chi2 = 5.272, P<0.05), UACS (2.2% (1/45), chi2 = 10.657, P<0.01) and GERC (0% (0/46), chi2 = 13.833, P<0.01). The specificity of nocturnal cough for CVA was 95.9%. The sensitivity and specificity of cough associated with meals in GERC was 52.2% (24/46) and 83.3%, and regurgitation associated symptom in GERC were 69.6% (32/46) and 80.0%, which were significantly higher than other groups. The incidence of postnasal drip, rhinitis associated symptom and case history of nasal diseases in UACS were 66.7% (30/45), 88.9% (40/45) and 82.2% (37/45), and the specificity of them were 89.4%, 65.6% and 63.6% respectively. CONCLUSION: The timing character and some associated symptoms of chronic cough are useful in predicting a single cause.
Subject(s)
Cough/diagnosis , Cough/etiology , Adolescent , Adult , Aged , Asthma/complications , Asthma/diagnosis , Chronic Disease , Diagnosis, Differential , Female , Gastroesophageal Reflux/complications , Gastroesophageal Reflux/diagnosis , Humans , Male , Middle Aged , Nose Diseases/complications , Nose Diseases/diagnosis , Rhinitis/complications , Rhinitis/diagnosis , Young AdultABSTRACT
Drug repurposing and/or repositioning is an alternative method to develop new treatment for certain diseases. Albendazole was originally developed as an anthelmintic medication, and it has been used to treat a variety of parasitic infestations. In this study, we investigated the antitumor effect of albendazole and putative action mechanism. Results showed that albendazole dramatically decreased the cell viability of SCC cell lines (SCC12 and SCC13 cells). Albendazole increased apoptosis-related signals, including cleaved caspase-3 and PARP-1 in a dose-dependent fashion. The mechanistic study showed that albendazole induced endoplasmic reticulum (ER) stress, evidenced by increase of CHOP, ATF-4, caspase-4, and caspase-12. Pretreatment with ER stress inhibitor 4-PBA attenuated albendazole-induced apoptosis of SCC cells. In addition, albendazole decreased the colony-forming ability of SCC cells, together with inhibition of Wnt/ß-catenin signaling. These results indicate that albendazole shows an antitumor effect via regulation of ER stress and cancer stemness, suggesting that albendazole could be repositioned for cutaneous SCC treatment.
Subject(s)
Albendazole/therapeutic use , Antineoplastic Agents/therapeutic use , Carcinoma, Squamous Cell/drug therapy , Skin Neoplasms/drug therapy , Albendazole/chemistry , Albendazole/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Carcinoma, Squamous Cell/pathology , Cell Line, Tumor , Endoplasmic Reticulum Stress/drug effects , Humans , Keratinocytes/drug effects , Keratinocytes/pathology , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Tunicamycin/pharmacologyABSTRACT
BACKGROUND: Skin keratinocytes participate actively in inducing immune responses when external pathogens are introduced, thereby contributing to elimination of pathogens. However, in condition where the excessive inflammation is occurred, chronic skin disease such as psoriasis can be provoked. OBJECTIVE: We tried to screen the putative therapeutics for inflammatory skin disease, and found that salvianolic acid A (SAA) has an inhibitory effect on keratinocyte inflammatory reaction. The aim of this study is to demonstrate the effects of SAA in poly(I:C)-induced inflammatory reaction in skin keratinocytes. METHODS: We pre-treated keratinocytes with SAA then stimulated with poly(I:C). Inflammatory reaction of keratinocytes was verified using real-time polymerase chain reaction, enzyme-linked immunosorbent assay and Western blot. RESULTS: When skin keratinocytes were pre-treated with SAA, it significantly inhibited poly (I:C)-induced expression of inflammatory cytokines including interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor-α, and CCL20. SAA inhibited poly(I:C)-induced activation of nuclear factor-κB signaling. And SAA also inhibited inflammasome activation, evidenced by decrease of IL-1ß secretion. Finally, SAA markedly inhibited poly(I:C)-induced NLRP3 expression. CONCLUSION: These results demonstrate that SAA has an inhibitory effect on poly(I:C)-induced inflammatory reaction of keratinocytes, suggesting that SAA can be developed for the treatment of inflammatory skin diseases such as psoriasis.
ABSTRACT
Cutaneous squamous cell carcinoma (SCC) is a common cancer that significantly decreases the quality of life. It is known that external stimulus such as ultraviolet (UV) radiation induces cutaneous SCC via provoking oxidative stress. NAD(P)H dehydrogenase 1 (NQO1) is a ubiquitous flavoenzyme that functions as a guardian against oxidative stress. However, the effect of NQO1 on cutaneous SCC is not clearly elucidated. In this study, we investigated the effect of NQO1 on cutaneous SCC cells using the recombinant adenoviruses that can upregulate and/or downregulate NQO1 expression. Overexpression of NQO1 resulted in significant decrease of cell proliferation and colony forming activity of SCC lines (SCC12 and SCC13 cells). By contrast, knockdown of NQO1 increased the cell proliferation and colony forming activity. Accordingly, the levels of proliferation-related regulators, such as Cyclin D1, Cyclin E, PCNA, SOX2, and p63, were decreased by the overexpression of NQO1, while those were increased by knockdown of NQO1. In addition, NQO1 affected the invasion and migration of SCC cells in a very similar way, with the regulation of epithelial-mesenchymal transition- (EMT-) related molecules, including E-cadherin, N-cadherin, Vimentin, Snail, and Slug. Finally, the overexpression of NQO1 decreased the level of phosphorylated AKT, JNK, and p38 MAPK, while the knockdown of NQO1 increased the level of phosphorylated signaling molecules. Based on these data, NQO1 has tumor suppressive function in cutaneous SCC cells.
Subject(s)
Carcinoma, Squamous Cell/genetics , Gene Expression Regulation, Neoplastic/genetics , NAD(P)H Dehydrogenase (Quinone) , Skin Neoplasms/genetics , Carcinoma, Squamous Cell/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/genetics , Epithelial-Mesenchymal Transition/drug effects , Epithelial-Mesenchymal Transition/genetics , Humans , NAD(P)H Dehydrogenase (Quinone)/genetics , NAD(P)H Dehydrogenase (Quinone)/metabolism , NAD(P)H Dehydrogenase (Quinone)/pharmacology , Signal Transduction/drug effects , Signal Transduction/genetics , Skin Neoplasms/metabolismABSTRACT
Three new labdane-type diterpenoids, callicapene M3-M5 (1-3) were isolated from the Callicarpa macrophylla Vahl. Their structures were identified by spectroscopic method. The isolated compounds were evaluated for inhibitory activity on NO production in LPS-activated RAW 264.7 macrophage cells by using MTT assays. Compounds 1-3 showed potent inhibitory activity, with IC50 value of 48.15, 46.31 and 38.72 µM respectively.
Subject(s)
Callicarpa/chemistry , Diterpenes/isolation & purification , Nitric Oxide/antagonists & inhibitors , Animals , Diterpenes/chemistry , Diterpenes/pharmacology , Inhibitory Concentration 50 , Macrophages/drug effects , Macrophages/metabolism , Mice , Molecular Structure , Nitric Oxide/biosynthesis , Plant Leaves/chemistry , RAW 264.7 CellsABSTRACT
Wilms tumor gene on the X chromosome (WTX) is a putative tumor suppressor gene in Wilms tumor, but its expression and functions in other tumors are unclear. Colorectal cancer (CRC) is the third leading cause of cancer-related deaths in women and the second leading cause in men in the United States. We demonstrated that WTX frequently lost in CRC which was highly correlated with cell proliferation, tumor invasion and metastasis. Mechanistically, WTX loss disrupts the interaction between RhoGDIα and CDC42 by losing of the binding with RhoGDIα and triggers the activation of CDC42 and its downstream cascades, which promotes CRC development and liver metastasis. The aberrant upregulation of miR-20a/miR-106a were identified as the reason of WTX loss in CRC both in vivo and in vitro. These study defined the mechanism how miR-20a/miR-106a-mediated WTX loss regulates CRC progression and metastasis, and provided a potential therapeutic target for preventing CRC progression.
Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Colonic Neoplasms/genetics , MicroRNAs/genetics , Tumor Suppressor Proteins/genetics , cdc42 GTP-Binding Protein/genetics , rho Guanine Nucleotide Dissociation Inhibitor alpha/genetics , Adaptor Proteins, Signal Transducing/metabolism , Animals , Cell Line, Tumor , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , Disease Progression , Female , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , HCT116 Cells , HT29 Cells , Humans , Male , Mice, Inbred BALB C , Mice, Nude , Middle Aged , Signal Transduction/genetics , Transplantation, Heterologous , Tumor Suppressor Proteins/metabolism , cdc42 GTP-Binding Protein/metabolism , rho Guanine Nucleotide Dissociation Inhibitor alpha/metabolismABSTRACT
OBJECTIVES: To review trials on the efficacy and safety of auricular acupuncture (AA) treatment for insomnia and to identify the most commonly used auricular acupoints for treating insomnia in the studies via a frequency analysis. DATA SOURCES: The international electronic databases searched included: (1) AMED; (2) the Cochrane library; (3) CINAHL; (4) EMBASE; and (5) MEDLINE. Chinese electronic databases searched included: (1) VIP Information; (2) CBMdisc; and (3) CNKI. STUDY SELECTION: Any randomized controlled trials using AA as an intervention without using any co-interventions for insomnia were included. Studies using AA versus no treatment, placebo, sham AA, or Western medicine were included. DATA EXTRACTION: Two (2) independent reviewers were responsible for data extraction and assessment. The efficacy of AA was estimated by the relative risk (RR) using a meta-analysis. RESULTS: Eight hundred and seventy eight (878) papers were searched. Six (6) trials (402 treated with AA among 673 participants) that met the inclusion criteria were retrieved. A meta-analysis showed that AA was chosen with a higher priority among the treatment subjects than among the controls (p < 0.05). The recovery and improvement rates produced by AA was significantly higher than those of diazepam (p < 0.05). The rate of success was higher when AA was used for enhancement of sleeping hours up to 6 hours in treatment subjects (p < 0.05). The efficacy of using Semen vaccariae ear seeds was better than that of the controls (p < 0.01); while magnetic pearls did not show statistical significance (p = 0.28). Six (6) commonly used auricular acupoints were Shenmen (100%), Heart (83.33%), Occiput (66.67%), Subcortex (50%), Brain and Kidney (each 33.33%, respectively). CONCLUSIONS: AA appears to be effective for treating insomnia. Because the trials were low quality, further clinical trials with higher design quality, longer duration of treatment, and longer follow-up should be conducted.
Subject(s)
Acupuncture Points , Acupuncture, Ear/methods , Sleep Initiation and Maintenance Disorders/therapy , Acupuncture, Ear/statistics & numerical data , Humans , Research Design , Sleep , Sleep Initiation and Maintenance Disorders/epidemiology , Treatment OutcomeABSTRACT
OBJECTIVE: To confirm the role of Tiam1 (T lymphoma invasion and metastasis 1) gene in the proliferation and metastasis of colorectal cancer. METHODS: Proliferative and metastatic abilities of Tiam1 transfectant were investigated by subcutaneous injection of cells and surgical orthotopic transplantation (SOI) in mice. RESULTS: The expression of Tiam1 led to a pronounced increase in HT29/Tiam1 cell growth starting from day 7, up to 2.5 fold increase of tumor volume at day 20 post injection. Tumors in the HT29/Tiam1 group receiving surgical orthotopic implantation were significantly heavier than those in HT29/mock group (t = -14.916, P < 0.01). In vivo metastasis assay by SOI showed that in HT29/Tiam1 group, 7/7 of mice developed peritoneal metastases and 4/7 had hepatic lesions. In addition, one of the seven HT29/Tiam1 group mice had tumors in lung, spleen and lymph nodes. In the HT29/mock group, only 2/7 of animals had peritoneal metastases and none produced detectable tumor in the liver. CONCLUSIONS: Tiam1 gene plays an important role in the proliferation, invasion and metastasis of colorectal cancer. It may serve as a useful clinical marker for tumor progression and metastasis of colorectal cancer.
Subject(s)
Cell Proliferation , Colorectal Neoplasms/pathology , Guanine Nucleotide Exchange Factors/metabolism , Peritoneal Neoplasms/secondary , Animals , Biomarkers, Tumor/analysis , Guanine Nucleotide Exchange Factors/genetics , Guanine Nucleotide Exchange Factors/physiology , HT29 Cells , Humans , Liver Neoplasms/secondary , Lung Neoplasms/secondary , Lymphatic Metastasis , Mice , Mice, Nude , Neoplasm Invasiveness , Neoplasm Transplantation , Plasmids , T-Lymphoma Invasion and Metastasis-inducing Protein 1 , Transfection , Tumor BurdenABSTRACT
WTX (Wilms' tumor suppressor X chromosome) is a novel putative tumor suppressor gene in Wilms' tumor of kidney, its expression and function in other human cancers had not been explored. This study detected the expression of WTX in 459 cases of 15 organs of cancers and adjacent normal tissues by using immunohistochemical staining (IHC), and validated them by in situ hybridization (ISH) and quantitative real-time reverse transcription PCR (qRT-PCR). IHC and ISH data showed that WTX protein was generally expressed in normal tissues, but reduced expression in corresponding cancers. This study demonstrated that WTX downregulation is a common phenomenon in human cancers, WTX might be a general tumor-suppressor gene and biological marker of multiple cancer tissues. Apart from kidney, stomach is another target tissue of WTX gene. The germline and somatic mutations of WTX were screened in 12 gastric cancer patients and identified in one cases (8.3%). Mutation in the WTX gene might be one of the reasons of WTX loss in gastric cancer patients.
Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Gene Expression/genetics , Neoplasms/genetics , Tumor Suppressor Proteins/genetics , Biomarkers, Tumor/genetics , Down-Regulation/genetics , Humans , Mutation/geneticsABSTRACT
OBJECTIVE: To test the effect of CDC42 (a member of Rho family of small GTPases) knockdown mediated by a CDC42 short-hairpin RNA (shRNA) on the morphology of colorectal cancer SW480 cells in vitro. METHODS: Four CDC42 siRNA fragments targeting CDC42 were designed and the most efficient siRNA for CDC42 knockdown was selected to construct the shRNA vector for transfection of colorectal cancer SW480 cells. The interference efficiency in the stably transfected cells (sw480.shCDC) was detected using real-time PCR and Western blotting, and the morphological changes of the transfected cells were observed. RESULTS: Western blotting result showed that siCDC42-3 was the most efficient fragment for CDC42 knockdown, which caused CDC42 knockdown by over 50%. DNA sequencing confirmed successful construction of the CDC42 shRNA vector. Transfection of the cells with the vector significantly reduced CDC42 expressions at both the mRNA and protein levels. The transfected cells exhibited reduced filopodia and cell size with smooth cell margins. CONCLUSION: shRNA-mediated CDC42 knockdown can reduce the cytoskeleton dynamics of colorectal cancer cells to lower their invasiveness. This shRNA construct facilitates further study of the role of CDC42 in the tumorigenesis and progression of colorectal cancer.
Subject(s)
Colorectal Neoplasms/genetics , RNA Interference , RNA, Small Interfering , cdc42 GTP-Binding Protein/genetics , Cell Line, Tumor , Gene Knockdown Techniques , Humans , RNA, Messenger , Real-Time Polymerase Chain Reaction , TransfectionABSTRACT
OBJECTIVE: To evaluate the effects of amino acids (AA) on the development of in vitro cultured preimplantation embryos of Kunming mice, and define the optimal AA concentration for embryo culture. METHODS: Totally 630 zygotes were collected from the oviducts of superovulated female Kunming mice, which were cultured in protein-free potassium simplex optimized medium (mKSOM) supplemented with Eagle's essential amino acids and Eagle's non-essential amino acids of different concentrations (mKSOM, mKSOM+1/16AA, mKSOM+1/8AA, mKSOM+1/4AA, mKSOM+1/2AA, mKSOM+AA, and mKSOM+2AA). RESULTS: The embryos cultured with the amino acids showed higher development rate to both 8-cell embryo stage and blastocyst stage than those cultured without amino acids. The correlation of amino acid concentration with 8-cell and blastocyst development rates conformed to the cubic model, with the highest development rate to both of the two stages observed at half of the amino acid concentration. CONCLUSION: Amino acids can promote the development of preimplantation Kunming mouse embryos, but excessively high concentration of amino acids impair embryo development possibly because of metabolic and osmotic pressure changes of the embryos as well as toxicity of ammonium resulting from the metabolism of amino acids.
Subject(s)
Amino Acids/pharmacology , Embryo, Mammalian , Embryonic Development/drug effects , Zygote/cytology , Animals , Culture Media , Dose-Response Relationship, Drug , Embryo, Mammalian/embryology , Female , Male , Mice , Organ Culture TechniquesABSTRACT
OBJECTIVE: To investigate the efficacy of posturography with inclinometer and platform with global bottom to make the subjects contact the ground almost at one point in the evaluation of body balance function. METHODS: The body balance function of 100 normal persons, 50 males and 50 females, aged 21 approximately 50, and 87 patients with peripheral vertigo due to semicircular canal palsy, aged 20 approximately 60, was measured with a new type of posturograph with inclinometer and platform with global bottom developed by the authors. The subjects with an inclination-sensor being fixed at the waist were asked to adopt four postures during the measurement: standing on the ground with eyes opened (test 1), standing on the ground with the eyes closed (test 2), standing on the platform with the eyes opened (test 3), and standing on the platform with the eyes closed (test 4). RESULTS: The measurement of the normal persons showed the average values of body sway angle velocity were 0.57 +/- 0.24 d/s in test 1, 0.70 +/- 0.18 d/s in test 2, 1.16 +/- 0.32 d/s in test 3, and 2.67 +/- 0.70 d/s in test 4. Sixty-two out of the 87 patients (71.26%) showed abnormality in one or more than one of the 4 tests with a value more than x +/- 2s of the normal value, among them 7 showed abnormal value in test 1, 14 in test 2, 16 in test 16, and 56 in test 4. Two weeks after the onset of vertigo, 55 out of the 87 patients showed abnormal values by this examination. CONCLUSION: Posturography with inclinometer and platform with global bottom is effective on evaluation of balance function, In particular, the test of standing on the platform with the eyes closed effectively reflects the vestibulospinal reflex.