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Objective:To investigate the clinical value of three-dimensional (3D) visualization technology in the precise drainage through endoscopic retrograde biliary drainage (ERBD) for hilar cholangiocarcinoma.Methods:Clinical data of 42 patients with highly suspected hilar cholangiocarcinoma who underwent ERBD in Qinghai University Affiliated Hospital from September 2019 to August 2022 were retrospectively collected. Twenty patients underwent 3D biliary tract reconstruction before surgery (the reconstruction group) and 22 others did not undergo 3D biliary tract reconstruction before surgery (the non-reconstruction group). The surgery time, X-ray exposure time, the technical success rate, the clinical success rate, incidence of postoperative complications, recent and short-term endoscopic retrograde cholangiopancreatography (ERCP) reintervention rate of the two groups were compared.Results:There was no significant difference in preoperative baseline data between the two groups ( P>0.05). ERBD was conducted successfully in all 42 patients. The operation time in the reconstruction group [35.00 (25.00, 57.50) min] was significantly shorter than that in the non-reconstruction group [60.00 (33.75, 60.00) min] with significant difference ( Z=-2.251, P=0.024). There was no significant difference in the X-ray exposure time between the two groups [10.00 (5.00, 12.00) min VS 10.55 (9.50, 17.50) min, Z=-1.552, P=0.121]. The technical success rates of both groups were 100.0%, and the clinical success rate of the reconstruction group was higher than that of the non-reconstruction group [70.0% (14/20) VS 31.8% (7/22)] with significant difference ( χ 2=6.109, P=0.013). There was no significant difference in the incidence of postoperative complications between the two groups [20.0% (4/20) VS 22.7% (5/22), χ 2=0.141, P=0.708]. All patients were followed up for 6 months after the procedure. The median survival time was 3.91 months in the reconstruction group and 2.78 months in the non-reconstruction group. There was no ERCP intervention in the reconstruction group within 2 weeks after the procedure, while 4 cases (18.2%) in the non-reconstruction group received 6 ERCP interventions due to cholangitis and postoperative pancreatitis. Within 2 weeks to 3 months, 2 patients (10.0%) in the reconstruction group received 4 ERCP interventions for cholangitis, and 2 patients (9.1%) in the non-reconstruction group received 3 ERCP interventions for cholangitis. There was no significant difference in recent ( χ 2=2.183, P=0.140) or short-term ( χ 2=0.000, P=1.000) ERCP reintervention rate between the reconstruction group and the non-reconstruction group. Conclusion:3D visualization biliary duct reconstruction technology can measure the volume of liver drainage for hilar cholangiocarcinoma, shorten the operation time and improve the clinical success rate through precise preoperative planning, which is worth of promotion.
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BACKGROUND@#The switch/sucrose nonfermentable chromatin-remodeling (SWI/SNF) complex is a pivotal chromatin remodeling complex, and the genomic alterations (GAs) of the SWI/SNF complex are observed in several cancer types, correlating with multiple biological features of tumor cells. However, their role in liver metastasis of non-small cell lung cancer (NSCLC) remains unclear. Our study aims to investigate the role and potential mechanisms underlying NSCLC liver metastasis induced by the GAs of SWI/SNF complex.@*METHODS@#The GAs of SWI/SNF complex in NSCLC cell lines (H1299, H23 and H460) were identified by whole-exome sequencing (WES). ARID1A knockout H1299 cell was constructed with the CRISPR/Cas9 technology. The mouse model of liver metastasis from NSCLC was established to simulate lung cancer liver metastasis and observe the metastasis rate under different gene mutation conditions. RNA sequencing and Western blot were conducted for differential gene expression analysis. Immunohistochemistry (IHC) analysis was used to assess protein expression levels of SWI/SNF-regulated target molecules in mouse liver metastases.@*RESULTS@#WES analysis revealed intracellular gene mutations. The animal experiments demonstrated a correlation between the GAs of SWI/SNF complex and a higher liver metastasis rate in immunodeficient mice. Transcriptome sequencing and Western blot analysis showed upregulated expression of ALDH1A1 and APOBEC3B in SWI/SNF-mut cells, particularly in ARID1A-deficient H460 and H1299 sgARID1A cells. IHC staining of mouse liver metastases further demonstrated elevated expression of ALDH1A1 in the H460 and H1299 sgARID1A group.@*CONCLUSIONS@#This study underscores the critical role of the GAs of SWI/SNF complex, such as ARID1A and SMARCA4, in promoting liver metastasis of lung cancer cells. The GAs of SWI/SNF complex may promote liver-specific metastasis by upregulating ALDH1A1 and APOBEC3B expression, providing novel insights into the molecular mechanisms underlying lung cancer liver metastasis.
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Animals , Mice , Carcinoma, Non-Small-Cell Lung/genetics , Lung Neoplasms/genetics , Mutation , Liver Neoplasms/geneticsABSTRACT
Sangzhi alkaloids (SZ-A) are derived from traditional Chinese medicine Ramulus Mori, serving well as an innovative antidiabetic drug, due to α-glucosidase inhibition. To evaluate the potency of glucosidase inhibitory effect of SZ-A, the enzyme-based screening platforms, including sucrase, maltase and amylase were established, and IC50 was calculated. The effects of SZ-A on postprandial blood glucose at a single dose, oral sucrose, starch and glucose loading were determined in normal ICR mice and alloxan-induced hyperglycemic mice. To confirm the anti-diabetic effects of SZ-A on glucose and lipid metabolism after long-term administration, the postprandial and fasting blood glucose, serum insulin, urinary glucose levels, glycosylated serum proteins and blood lipid levels were determined in high-fat fed C57 obese mice (pre-diabetic HFC57 mice) and diabetic rats induced by streptozotocin (STZ). The Experimental Animal Welfare Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College approved all of the protocols for this research. We found that SZ-A exhibited a significant inhibitory effect on the sucrase and maltase. SZ-A showed no effect on amylase. In normal ICR mice and alloxan-induced hyperglycemic mice, SZ-A at a single dose significantly delayed and reduced the peak of blood glucose after sucrose or starch loading, but showed no effect on the increase of blood glucose after glucose loading. In STZ diabetic rats, SZ-A significantly reduced the postprandial or fasting blood glucose levels, glycosylated serum proteins and urinary glucose. SZ-A also reduced serum triglyceride (TG) and cholesterol (TC) levels after 3 weeks of treatment. SZ-A ameliorated the postprandial blood glucose or the fasting blood glucose elevation, and reduced the incidence of hyperglycemia in HFC57 mice. SZ-A decreased the basal insulin level, improved insulin sensitivity, and ameliorated glucose intolerance in pre-diabetic HFC57 mice. Our results indicated that SZ-A had a novel inhibitory activity on α-glucosidase, especially on disaccharidases. SZ-A at a single dose significantly reduced the peak of blood glucose elevation and delayed the increase of blood glucose in normal and diabetic mice after disaccharide and polysaccharide loading. Long-term SZ-A treatment improved glucose and lipid metabolic profiles by delaying carbohydrate absorption from the intestine and reduced the postprandial blood glucose levels in both pre-diabetic and diabetic animal models. Therefore, SZ-A application may display a beneficial role in preventing the development and complications of diabetes.
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<p><b>BACKGROUND</b>SH3TC2, PMP2, and BSCL2 genes are related to autosomal recessive (AR) Charcot-Marie-Tooth (CMT) disease type 1, autosomal dominant (AD)-CMT1, and AD-CMT2, respectively. Pathogenic variants in these three genes were not well documented in Chinese CMT patients. Therefore, this study aims to detect SH3TC2, PMP2, and BSCL2 pathogenic variants in a cohort of 315 unrelated Chinese CMT families.</p><p><b>METHODS</b>A total of 315 probands from 315 unrelated Chinese CMT families were recruited from the Department of Neurology of Third Xiangya Hospital and Xiangya Hospital. We screened for SH3TC2 pathogenic variants in 84 AR or sporadic CMT probands, PMP2 pathogenic variants in 39 AD or sporadic CMT1 probands, and BSCL2 pathogenic variants in 50 AD or sporadic CMT2 probands, using polymerase chain reaction and Sanger sequencing. All these patients were out of 315 unrelated Chinese CMT families and genetically undiagnosed after exclusion of pathogenic variants of PMP22, MFN2, MPZ, GJB1, GDAP1, HSPB1, HSPB8, EGR2, NEFL, and RAB7. Candidate variants were analyzed based on the standards and guidelines of American College of Medical Genetics and Genomics (ACMG). Clinical features were reevaluated.</p><p><b>RESULTS</b>We identified three novel heterozygous variants such as p.L95V (c.283C>G), p.L1048P (c.3143T>C), and p.V1105M (c.3313G>A) of SH3TC2 gene and no pathogenic variants of PMP2 and BSCL2 genes. Although evaluation in silico and screening in the healthy control revealed that the three SH3TC2 variants were likely pathogenic, no second allele variants were discovered. According to the standards and guidelines of ACMG, the heterozygous SH3TC2 variants such as p.L95V, p.L1048P, and p.V1105M were considered to be of uncertain significance.</p><p><b>CONCLUSIONS</b>SH3TC2, PMP2, and BSCL2 pathogenic variants might be rare in Chinese CMT patients. Further studies to confirm our findings are needed.</p>
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AIM:To predict and identify an HLA-A3 supertype-restricted cytotoxic T-lymphocyte(CTL) epitope derived from MAGEC2,which is utility in epitope design for the development of HLA-based vaccines and immuno-therapeutics.METHODS:HLA-A3 epitopes from MAGEC2 protein were predicted by BIMAS, SYFPEITHI and IEDB. The binding affinity of the peptides to HLA-A*03 molecule was evaluated by T2A3 cell binding assay.ELISPOT assay was used to investigate the ability of the peptides inducing specific restricted CTLs to release interferon -γ(IFN-γ).The ability of the peptides to induce T-cell response was investigated by cytotoxicity assay in vitro.RESULTS:The candidate peptides P147,P167, P196, P229 and P251 showed moderate affinity toward HLA-A3 molecule.ELISPOT assay showed that P167,P196 and P251 were able to induce specific CTLs and higher levels of IFN-γwere released.The CTLs induced by P196 and P251 were able to lyse target cells.CONCLUSION:The peptides P196 and P251 have higher binding affinity with HLA-A3 and retain immunogenicity.They are excellent HLA-A3-restricted CTL epitopes from tumor antigen MA-GEC2,which could serve as new candidates towards antitumor peptide vaccines.
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Objective To investigate the impact of fasting blood glucose on the prognosis of non-diabetic patients undergoing primary percutaneous coronary intervention (PCI) for acute ST-segment elevation myocardial infarction (STEMI). Methods We retrospectively recruited consecutive patients who underwent primary PCI in our hospital from February, 2003 to March, 2015. Patients with prior history of diabetes mellitus before the index hospitalization and those with newly diagnosed diabetes mellitus during the index hospitalization were excluded. The clinical and angiographic features, medical and interventional treatment, and 30-day outcomes were compared between patients with elevated fasting blood glucose (FBG) (>5.4 mmol/L) and those with normal FBG (≤5.4 mmol/L). Results A total of 721 patients were recruited with an age of(61.2 ± 12.8)years, of whom 601 (83.4 %) were male. As compared with patients with normal FBG,those with elevated FBG were more likely to be female(20.1 % vs.13.5 %, P=0.017),had faster heart rate on admission[(82.9 ± 17.2)bpm vs.(79.4 ± 16.7)bpm,P=0.006]and more use of intra-aortic balloon pump(3.8 % vs.1.3 %,P=0.034),and had higher rates of 30 day all-cause mortality(3.5 % vs.0.5 %,P=0.004),cardiac mortality(2.9 % vs.0.5 %,P=0.012)and heart failure(18.1 % vs. 7.4 %, P< 0.001). After adjusting baseline characteristics, FBG > 5.4 mmol/L was one of the independent predictors of 30-day all-cause mortality(HR 6.030,95 % CI 1.235-29.447,P=0.026).Other independent predictors of 30-day all-cause mortality included age(HR 1.059,95 % CI 1.002-1.120,P=0.044),heart rate on admission(HR 1.036,95 % CI 1.003-1.070,P=0.034),left descending artery as the culprit vessel(HR 6.427,95 % CI 1.389-29.728,P=0.017),and use of angiotensin converting enzyme inhibitor/angiotensin receptor blocker(HR 0.154,95 % CI 0.051-0.461,P=0.001).Conclusions In non-diabetic patients undergoing primary PCI for STEMI, elevated FBG was one of the independent predictors of 30-day all-cause mortality.
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Objective To analyze the characteristics, treatment and prognosis during follow-up in patients with essential thrombocythemia(ET)after percutaneous coronary intervention(PCI). Methods A total of 9 patients with ET who had CAD and treated with PCI in our hospital from 2010-01 to 2017-07 were retrospectively studied. The basic clinical information with clinical data,data of coronary intervention,application of anti-platelet and platelet reducing drugs,and the results of long-term follow-up were recorded in all patients. Results There were 5 male and 4 female with a mean age of (60.9 ± 14.3) years The 9 patients included 3 cases of acute myocardial infaction(MI),3 cases of old MI,2 cases of unstable angina and 2 cases stable angina. Coronary angiography revealed that there were 4 cases with single vessle involved and 5 cases with two-three vessles involved. All the patients were implante with drug-eluting stents. The mean platelet counts of the patients at admission were(596±233)×109/L. In 4 patients, ET was diagnosed during current hospitalization and their mean platelet counts were(803±105)×109/L. All patients received dual anti-platelet drugs after PCI and 8 patients had hydroxyurea during follow-up. During the follow-up of (53.2±26.9) months,no cardiovascular events occurred. One case of gastrointestinal bleeding occurred during the follow-up due to gastric ulcer. Conclusions Application of platelet reducing therapy with dual anti-platelet drugs in ET patients after PCI is eff ective and safe.
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Aim To assess the effects of Trillium Tschonoskii Maxim ( TTM ) decoction on Tau protein phosphorylation and synaptic development in AD model rats induced by high activity GSK-3β. Methods The SD rats were divided into five groups of ten animals, named sham-operated group ( blank group) , AD model group, TTM group (0. 5, 0. 25, 0. 125 g·kg-1 · d-1 ) . Treatment group received gavage once a day for seven days with TTM decoction, while other groups by gavage once a day for seven days with drinking water. On 2nd day by gavage, Morris water maze test was used to assess the spatial learning and memory ability of the rats. After five days' training, rats in the treat-ment groups and AD model group were injected wort-mannin ( WT, PI3K specific inhibitor ) and GF-109203X (GFX, PKC specific inhibitor) (100 μmol ·L-1 of each, total volume of 10 μL) into the right lateral ventricle. Western blot was used to detect the levels of phosphorylation Tau protein at multiple sites and the expression level of PI3K, Akt, PKC, GSK-3β(S9, T216) and synapse-associated proteins. Immu-nohistochemical method was used to detect the hyper-phosphorylation of Tau protein in hippocampus of rats. Golgi staining was applied to detect the number and morphological changes of synaptic development and dendritic spines. Nissl' s staining was employed to ob-serve the development of neonatal neurons in hippo-campus and cortex. Results Western blot showed that the phosphorylation level of Tau in hippocampus increased in model group, and the activity of GSK-3βwas up-regulated. Among them, however, in middle dose TTM group, the phosphorylation level of Tau in hippocampus decreased and the activity of GSK-3βde-creased. The expression levels of p-PKC and p-Akt in low and middle dose treatment group were higher than those in model group, thus increasing the activity of PKC and Akt to inhibit the activity of GSK-3β kinase. Immunohistochemistry also indicated that TTM could decrease the biological effects of Tau phosphorylation in hippocampus of AD rats. Western blot showed that TTM could increase the expression levels of synapsin-1 , syn-aptophysin and GluR-1 in hippocampus of AD rats. Nissl staining showed that the number of Nissl bodies in hippocampal neurons of AD model group were signif-icantly fewer than those of sham operation group, which could be increased by TTM middle and high dose group, and the complexity and dendritic spine density of hippocampal neurons in AD rats could be en-hanced as well. Conclusion TTM can effectively im-prove the cognitive function of AD rats induced by the increase of GSK-3β activity, and its possible mecha-nism may be via down-regulating the activity of GSK-3β and inhibiting the phosphorylation of tau protein and promoting the development of neurons.
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Aim To assess the effects of Trillium Tschonoskii Maxim ( TTM) decoction on learning and memory dysfunction in Alzheimer’ s disease ( AD ) model rats which induced by okadaic acid( OA) and its possible mechanism. Methods The SD rats were di-vided into ten groups,namely,d DMSO control group, OA group, TTM high-dose ( 0. 5 g·kg-1·d-1) group,TTM medium-dose ( 1 g·kg-1·d-1) group, TTM lower-dose (2 g·kg-1·d-1) group,and these groups were divided into one week and two weeks of gavage. Treatment groups were gavaged with TTM de-coction twice a day. After 5 days of Morris water maze training,treatment groups and AD model groups were injected with OA (0.392 mmol·L-1,1. 5 μL) in bi-lateral hippocampal of the rats. The DMSO groups were injected with 10% DMSO. The spatial memory reten- tion wereas detected by water maze at 24 h after injec-tion. After the test, we prepared sample for Western blot and Nissl’s staining. The Western blotting test was used to detect the PP2A activity and the phospho-rylation of Tau protein in the hippocampus. Nissl’'s staining was used to observe the changes of the number of Nissl’s bodies in the hippocampal CA1 and CA3 re-gions. Results The Morris water maze test showed that after injection of OA, the latency of TTM groups wereas shorter than that of OA groups. Western blot showed that the high dose TTM could increase the ac-tivity of PP2A and decrease the level of Tau phospho-rylation at PS-Tau396,,PT-Tau404. The Nissl’s stai-ning results showed that the number of Nissl’s bodies in the hippocampal CA1 and CA3 regions of OA groups wereas significantly attenuated compared with that of the number of Nissl's bodies in the hippocampal CA1 and CA3 regions than DMSO groups. The number of Nissl’s bodies in high groups were morewas larger than that of OA group. Conclusion The results show that TTM can improve the learning and memory dysfunction in AD model rats which induced by OA. The mecha- nism wasis probably that TTM can increase PP2A ac-tivity and then down-regulate the level of Tau phospho-rylation and improve neural development.
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Advances in biological and medical technologies have been providing us explosive volumes of biological and physiological data, such as medical images, electroencephalography, genomic and protein sequences. Learning from these data facilitates the understanding of human health and disease. Developed from artificial neural networks, deep learning-based algorithms show great promise in extracting features and learning patterns from complex data. The aim of this paper is to provide an overview of deep learning techniques and some of the state-of-the-art applications in the biomedical field. We first introduce the development of artificial neural network and deep learning. We then describe two main components of deep learning, i.e., deep learning architectures and model optimization. Subsequently, some examples are demonstrated for deep learning applications, including medical image classification, genomic sequence analysis, as well as protein structure classification and prediction. Finally, we offer our perspectives for the future directions in the field of deep learning.
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Humans , Algorithms , Computational Biology , Methods , Diagnostic Imaging , Genomics , Methods , Image Interpretation, Computer-Assisted , Methods , Machine Learning , Neural Networks, Computer , Protein Structure, Secondary , Proteins , MetabolismABSTRACT
Objective: To explore the prevalence and risk factors of orthostatic hypotension (OH) in elderly hypertension patients. Methods: A total of 532 retired hypertension patients elder than 65 years in Guangzhou military region were enrolled. The patients were divided into 2 groups: Hypertension group, n=414 and Hypertension combining OH (H+OH) group, n=118. The patient's age (65-79、≥ 80), hypertension grade (Grade 1-3) and complication status were studied. The risk factors for H+OH prevalence were analyzed by multivariate Logistic regression analysis. Results: The incidence rate of H+OH was 22.2％ (118/532). In H+OH group, the ratios of elderly and very elderly patients were 6.7％ and 23.1％, P<0.05 and the ratios of OH occurrence for hypertension grade 1, 2 and 3 were 12.6％, 23.3％ and 25.2％ respectively, P<0.05. Multivariate Logistic regression analysis presented that systolic blood pressure (BP) in supine position, BP at immediate standing, heart rate in supine position, heart rate after 2 minutes standing and chronic cardiac insufficiency were the impact factors for H+OH occurrence, P<0.05. Conclusion: In elderly hypertension patients, incidence of OH was increasing with age elevating; H+OH has been related to age, severity of hypertension and chronic cardiac insufficiency.
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Objective To observe the effects of acupuncture treatment on the levels of HPA axis related hormones and acupoint compatibility in insomnia rats; To discuss the mechanism of action for acupuncture treatment. Methods Chlorophenylalanine suspension was under intraperitoneal injection to establish insomnia model rats. Sixty SD rats were randomly divided into blank group, model group, Baihui+Shenmen group, Baihui+Sanyinjiao group, Baihui+non-acupoint group, with 12 rats in each group. Each treatment group received acupuncture in relevant acupoints, 30 min each time, for 7 d. ELISA was used to measure the levels of CRH, ACTH and CORT. Results Compared with the model group, the levels of CRH, ACTH and CORT of the acupuncture groups decreased to some extent. In the three acupuncture groups, the efficacy of Baihui+Shenmen group was better than that of Baihui+Sanyinjiao group and Baihui+non-non-acupoint group. Conclusion Acupuncture treatment may calm and soothe the nerves to release the insomnia through regulating HPA axis related hormones. Acupuncture acupoints at different meridians may be one of the factors that cause the difference of acupoints compatibility effect.
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Objective: To explore the prevalence and risk factors of orthostatic hypotension (OH) in elderly hypertension patients. Methods: A total of 532 retired hypertension patients elder than 65 years in Guangzhou military region were enrolled. The patients were divided into 2 groups: Hypertension group, n=414 and Hypertension combining OH (H+OH) group, n=118. The patient's age (65-79、≥ 80), hypertension grade (Grade 1-3) and complication status were studied. The risk factors for H+OH prevalence were analyzed by multivariate Logistic regression analysis. Results: The incidence rate of H+OH was 22.2％ (118/532). In H+OH group, the ratios of elderly and very elderly patients were 6.7％ and 23.1％, P<0.05 and the ratios of OH occurrence for hypertension grade 1, 2 and 3 were 12.6％, 23.3％ and 25.2％ respectively, P<0.05. Multivariate Logistic regression analysis presented that systolic blood pressure (BP) in supine position, BP at immediate standing, heart rate in supine position, heart rate after 2 minutes standing and chronic cardiac insufficiency were the impact factors for H+OH occurrence, P<0.05. Conclusion: In elderly hypertension patients, incidence of OH was increasing with age elevating; H+OH has been related to age, severity of hypertension and chronic cardiac insufficiency.
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Objective To observe the effects of acupuncture treatment on the levels of HPA axis related hormones and acupoint compatibility in insomnia rats; To discuss the mechanism of action for acupuncture treatment. Methods Chlorophenylalanine suspension was under intraperitoneal injection to establish insomnia model rats. Sixty SD rats were randomly divided into blank group, model group, Baihui+Shenmen group, Baihui+Sanyinjiao group, Baihui+non-acupoint group, with 12 rats in each group. Each treatment group received acupuncture in relevant acupoints, 30 min each time, for 7 d. ELISA was used to measure the levels of CRH, ACTH and CORT. Results Compared with the model group, the levels of CRH, ACTH and CORT of the acupuncture groups decreased to some extent. In the three acupuncture groups, the efficacy of Baihui+Shenmen group was better than that of Baihui+Sanyinjiao group and Baihui+non-non-acupoint group. Conclusion Acupuncture treatment may calm and soothe the nerves to release the insomnia through regulating HPA axis related hormones. Acupuncture acupoints at different meridians may be one of the factors that cause the difference of acupoints compatibility effect.
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<p><b>OBJECTIVE</b>To explore the correlation of c-met protein with the clinical staging and cell differentiation of esophageal squamous cell carcinoma (ESCC).</p><p><b>METHODS</b>A total of 100 patients with ESCC were enrolled were examined for expression of c-met protein using immunohistochemistry, and the patients in negative and positive c-met expression groups were compared for clinicopathological characteristics and overall survival.</p><p><b>RESULTS</b>s The 100 ESCC patients included 67 male and 33 female patients with a median age of 59 years; 49 of the patients were negative and 51 were positive for c-met expression. Positive c-met expression was significantly correlated with advanced TMN stages and lower tumor differentiation. Kaplan-Meier survival curve showed that the median survival time of c-met-positive patients was significantly reduced compared with that of c-met-negative patients (30.9 vs 48.2 months, P<0.05). COX regression analysis showed that c-met was a independent risk factor for the overall survival of the patients (HR: 2.34, 95% CI: 1.63-4.54, P<0.05).</p><p><b>CONCLUSION</b>A positive expression of c-met protein is significantly correlated with an advanced TMN stage, lower tumor differentiation and a poor prognosis, and may serve as a indicator for predicting the prognosis of ESCC.</p>
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Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Diagnosis , Metabolism , Esophageal Neoplasms , Diagnosis , Metabolism , Immunohistochemistry , Kaplan-Meier Estimate , Neoplasm Staging , Prognosis , Proportional Hazards Models , Proto-Oncogene Proteins c-met , Metabolism , Risk FactorsABSTRACT
Objective To evaluate regulatory effects of glutamate receptor antagonists on the proliferation and migration of WM451LU malignant melanoma cells, and to explore their related mechanisms. Methods WM451LU cells at exponential growth phase were classified into 3 groups to be treated with the glutamate receptor antagonist MK?801 at 100μmol/L(MK?801 group), the glutamate receptor antagonist CPCCOEt at 10μmol/L(CPCCOEt group), or culture medium(control group). After 24?hour treatment, methyl thiazolyl tetrazolium(MTT)assay was performed to determine cell proliferation rates, scratch assay to evaluate the migration activity of cells, and Western?blot analysis to measure expression levels of proliferating cell nuclear antigen (PCNA), protein kinase Cα(PKCα) both on cell membrane and in cytoplasm, and phosphorylated mitogen?activated protein kinase(p?MAPK). Results After 24?hour treatment, cell proliferation rates were significantly decreased in the MK?801 group and CPCCOEt group compared with the control group(63%± 3.1%and 60%± 2.4%vs. 100%± 1.1%, both P<0.05). The scratch assay showed that cell?free zones in the control group gradually narrowed over time, and the scratch wound tended to close. However, the cell?free zones in the MK?801 group and CPCCOEt group narrowed more slowly compared with the control group, and were still wide after 24?hour culture with no obvious closure of the scratch. The MK?801 group and CPCCOEt group both showed significantly decreased expressions of PCNA(77.0% ± 5.4% and 72.0% ± 4.2% respectively), PKCα on the cell membrane(0.12 ± 0.02 and 0.14 ± 0.02 respectively), and p?MAPK(0.48 ± 0.03 and 0.36 ± 0.04 respectively) compared with the control group(PCNA:100.0%± 1.3%;PKCα:0.38 ± 0.01;p?MAPK:1.00 ± 0.02;all P<0.05).Conclusion In vitro suppression of glutamate receptors can inhibit the proliferation and migration of WM451LU cells, likely through the mediation of the PKCα?MAPK signaling pathway.
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AIM:Increasing evidence indicates that inflammation contributes to the initiation and perpetuation of atrial fibrillation ( AF) .Al-though tumor necrosis factor ( TNF)-αlevels are increased in patients with AF , the role of TNF-αin the pathogenesis of AF remains unclear.Recent research has revealed that T-type Ca2+currents ( ICa,T ) play an important role in the pathogenesis of AF .METH-ODS:In this study , we used the whole-cell voltage-clamp technique and biochemical assays to explore the role of TNF-αin the regula-tion of ICa,T in atrial myocytes.RESULTS:We found that compared with sinus rhythm (SR) controls, T-type calcium channel (TCC) subunit mRNA levels were decreased , while TNF-αexpression levels were increased , in human atrial tissue from patients with AF .In murine atrial myocyte HL-1 cells, after cultured for 24 h, 12.5, 25 and 50 μg/L TNF-αsignificantly reduced the protein expression levels of the TCC α1G subunit in a concentration-dependent manner .The peak current was reduced by the application of 12.5 or 25μg/L TNF-αin a concentration-dependent manner [from ( -15.08 ±1.11) pA/pF in controls to ( -11.89 ±0.83) pA/pF and (-8.54 ±1.55) pA/pF in 12.5 and 25 μg/L TNF-αgroups, respectively].TNF-αapplication also inhibited voltage-dependent inactivation of ICa,T shifted the inactivation curve to the left .CONCLUSION:These results suggest that TNF-αis involved in the path-ogenesis of AF, probably via decreasing ICa,T function in atrium-derived myocytes through impaired channel function and down -regula-tion of channel protein expression .This pathway thus represents a potential pathogenic mechanism in AF .
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Objective To develop neutralizing monoclonal antibodies (MAbs) against H10N8 avian influenza virus hemagglutinin and to identify the binding sites. Methods MAbs against hemagglutinin of H10N8 avian influenza virus were developed by genetic engineering. Neutralizing MAbs were screened by microneutralization assay,and then tested by enzyme-linked immunosorbent assay and Western blot to identity the binding sites.The homology modeling process was performed using Discovery Studio 3.5 software,while the binding epitopes were analyzed by BioEdit software. Results One MAb that could neutralize the H10N8 pseudovirus was obtained and characterized. Analysis about epitopes suggested that the antibody could bind to the HA1 region of hemagglutinin,while the epitopes on antigen were conserved in H10 subtypes.Conclusions One neutralizing antibody was obtained by this research.The MAb may potentially be further developed as a pre-clinical candidate to treat avian influenza H10N8 virus infection.
Subject(s)
Antibodies, Monoclonal , Allergy and Immunology , Antibodies, Neutralizing , Allergy and Immunology , Antibodies, Viral , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Epitopes , Allergy and Immunology , Hemagglutinin Glycoproteins, Influenza Virus , Allergy and Immunology , Influenza A Virus, H10N8 Subtype , Neutralization TestsABSTRACT
<p><b>OBJECTIVE</b>To study the distribution of mutations of UDP-glucuronosyltransferase 1A1 (UGT1A1) gene and its relationship with hyperbilirubinemia among neonates with hyperbilirubinemia of Guangxi Heiyi Zhuang nationality.</p><p><b>METHODS</b>Total genomic DNA was extracted from the blood of 100 neonates with hyperbilirubinemia (case group) and 100 neonates without hyperbilirubinemia (control group), all of whom were selected from Guangxi Heiyi Zhuang population. TATA box and all exons of UGT1A1 gene were amplified by PCR and directly sequenced.</p><p><b>RESULTS</b>(TA)7 insertion mutation in TATA box, G71R missense mutation in exon 1, and 4 single nucleotide polymorphisms (SNPs) (rs199539868, rs114982090, rs1042640 and rs8330) in exon 5 were observed. The allele frequency of G71R mutation in the case group was significantly higher than that in the control group (P<0.01). There were no significant differences in the genotype distribution and allele frequency of TATA box mutation and SNPs (rs1042640 and rs8330) between the two groups (P>0.05). The logistic regression analysis showed that the odds ratios (95% confidence intervals) of UGT1A1 TATA box mutation, G71R mutation, and SNPs (rs1042640 and rs8330) associated with the development of neonatal hyperbilirubinemia were 0.846 (0.440, 1.629), 3.932 (1.745, 8.858), 0.899 (0.364, 2.222), respectively.</p><p><b>CONCLUSIONS</b>(TA)7 insertion mutation and G71R missense mutation of UGT1A1 gene are common mutation types in neonates with hyperbilirubinemia of Guangxi Heiyi Zhuang nationality. Four SNPs (rs199539868, rs114982090, rs1042640, and rs8330) was first reported in China. UGT1A1 G71R missense mutation is a risk factor for hyperbilirubinemia in neonates of Guangxi Heiyi Zhuang nationality.</p>
Subject(s)
Humans , Infant, Newborn , China , Ethnology , Glucuronosyltransferase , Genetics , Hyperbilirubinemia, Neonatal , Genetics , Logistic Models , Mutation , Polymorphism, Single Nucleotide , TATA BoxABSTRACT
<p><b>OBJECTIVE</b>To express and purify recombinant human interleukin-35[IL-35-IgG1 (Fc) in eukaryotic expression system and to study the interaction of IL-35 with gp130 protein.</p><p><b>METHODS</b>A mammalian expression vector, pSTEP2-IL35-LFc, was constructed and transfected into HEK293T cells. Then rhIL-35-IgG1 (Fc) was expressed and purified with protein A affinity chromatography, and was examined with SDS-PAGE and Western blot analysis. The binding of IL-35 to its receptor gp130 was investigated using enzyme-linked immunosorbent assay (ELISA). The biological effect of IL-35 on gp130 was explored in M1 myeloid leukemia cells.</p><p><b>RESULTS</b>rhIL-35-Fc with high purity on reduced SDS-PAGE was obtained. ELISA confirmed that IL-35-Fc was bound to gp130 and neutralized the function of gp130 in M1 myeloid leukemic cells.</p><p><b>CONCLUSIONS</b>High-purity and biologically active rhIL-35-Fc protein successfully produced in this study. IL-35 binds to gp130 and neutralizes its activity of in M1 myeloid leukemic cells.</p>