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BACKGROUND@#LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer.@*METHODS@#We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels.@*RESULTS@#On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]).@*CONCLUSION@#LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile.@*TRIAL REGISTRATION@#ClinicalTrials.gov, NCT04563936.
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Humanos , Masculino , Antineoplásicos Hormonais/uso terapêutico , População do Leste Asiático , Hormônio Liberador de Gonadotropina/agonistas , Gosserrelina/uso terapêutico , Antígeno Prostático Específico , Neoplasias da Próstata/tratamento farmacológico , TestosteronaRESUMO
Objective: To explore the safety and feasibility of gasless submental approach endoscopic removal of thyroglossal cyst. Methods: This work prospectively included the clinical data of 13 patients who underwent the gasless submental approach endoscopic removal of thyroglossal cyst at the Department of Otolaryngology, the First Affiliated Hospital of Chongqing Medical University from August 2021 to February 2022. The operative time, bleeding volumes, postoperative complications, subjective pain levels, satisfaction with incisional scars, neck and facial deformities, and recurrences were prospectively evaluated by visual analogue scale(VAS) score. Results: All of 13 patients were successfully treated using this new technique. The patients had a median age of 38 years, ranging from 24 to 59 years, comprising of 3 males and 10 females. The length of the submental incision was about 3 cm and the median of operation time was 135 minutes. Postoperative complications were minimal. The median volume of blood loss was 10 ml. Surgical site swelling existed in 1 case and dysphagia for more than 1 week occurred in 2 cases. Patients were discharged from the hospital with a median of 3 days after surgery. On a VAS of 0-10 scores, the pain had a median of 2 on the first day after surgery, and the satisfaction with incision scars and neck and facial deformities showed a median of 8 at 6 months after surgery. There were no recurrences during the follow-up period of 9-15 months. Conclusion: Gasless submental approach endoscopic removal of thyroglossal cyst may be a reliable new surgical method that is safe and has cosmetic advantages.
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Masculino , Feminino , Humanos , Adulto , Cicatriz/complicações , Cisto Tireoglosso/complicações , Endoscopia/métodos , Complicações Pós-Operatórias , Dor/complicaçõesRESUMO
OBJECTIVE To observe whether mitochondria can be transferred from mesenchymal stem cells(MSCs)to irradiated cells by establishing a mitochondrial fluorescent reporting system.METHODS The lentiviral vector pSIN-EF1α-COX8A-DsRed2(named COX8A-DsRed2)that might guide the expres-sion of red fluorescence protein in the membrane of mitochondria was constructed.A lentivirus(named Lv-COX8A-DsRed2)was prepared in 293T cell line.Dental pulp stem cells(DPSCs)(named DPSC-COX8A-DsRed2)was infected with Lv-COX8A-DsRed2.The intracellular expression of the red fluores-cence protein in DPSC was observed under fluorescence microcopy.The mitochondrial localization of the expressed red fluorescent probe in DPSC-COX8A-DsRed2 was confirmed according to TOMM20 immunostaining and MitoTracker Green staining results,which could specifically label mitochondria.The IEC-6 cells that received 10 Gy X-ray radiation were used as an injured cell model.The co-culture system was established by supplementing DPSC-COX8A-DsRed2 into the culture plate with the irradi-ated IEC-6 labelled by CFSE for 24 h.RESULTS The imaging results of fluorescent microcopy obser-vation showed that DPSC-COX8A-DsRed2 expressed the mitochondrial fluorescent reporting system,which was co-located with TOMM20 protein and Mito Tracker Green.The imaging results of confocal fluorescence microcopy showed that the mitochondria with red fluorescent protein were transferred from DPSC-COX8A-DsRed2 to the irradiated IEC-6 cells,suggesting that the established mitochondrial fluorescent reporting system could indicate mitochondrial transfer from donor cells to injured ones.CONCLUSION DPSC-COX8A-DsRed2 stably expressing the mitochondrial fluorescent reporting system is established,which can be used to track mitochondrial transfer.
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Objective:To evaluate the clinical significance of neoadjuvant immunotherapy combined with chemotherapy in the treatment of larynx preservation in locally advanced hypopharyngeal squamous cell carcinoma. Methods:Patients with locally advanced HPSCC(cT3-T4aN0-N3M0) were eligible. All received 2 cycles of pembrolizumab combined with docetaxel and platinum neoadjuvant induction therapy. After two cycles, the efficacy was evaluated, followed by radical chemoradiotherapy or surgery and adjuvant chemoradiotherapy according to the efficacy. The primary endpoints were objective response rate(ORR) ,larynx-preservation(LP) rate at 3 months post-treatment and the adverse reactions during neoadjuvant therapy. Results:From December 2021 to December 2022, 10 patients with locally advanced HPSCC(cT3-T4aN0-N3M0) were enrolled. After 2 cycles of the neoadjuvant therapy, 2 patients achieved complete response(CR), 7 patients achieved partial response(PR), 1 patient was stable disease(SD), objective response rate(ORR) was 90%, and disease control rate(DCR) was 100%. 5 patients received radical chemoradiotherapy, 5 patients received surgery and adjuvant chemoradiotherapy, four of them received partial laryngectomy and partial hypopharyngeal resection surgery, and one of them received total laryngectomy and partial hypopharyngeal resection surgery. All patients were able to withstand adverse reactions of neoadjuvant therapy and successfully completed the whole treatment of HPSCC without grade 3-4 treatment-related adverse reactions. There was no recurrence or metastasis during 3-18 months of follow-up. 1 patient died of severe pneumonia 3 months after the completion of radical chemoradiotherapy. At 3 months after treatment, the larynx-preservation rate was 80%. Conclusion:Neoadjuvant immunotherapy combined with chemotherapy has good short-term efficacy and the adverse reactions were tolerable. It can improve the larynx-preservation rate of patients with locally advanced HPSCC, thus improving the prognosis and quality of life of patients.
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Humanos , Carcinoma de Células Escamosas de Cabeça e Pescoço/etiologia , Terapia Neoadjuvante , Qualidade de Vida , Cisplatino , Resultado do Tratamento , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Laringe , Neoplasias de Cabeça e Pescoço , ImunoterapiaRESUMO
Objective:To investigate the prognostic impact of different tumor invasion patterns in the surgical treatment of T3 glottic laryngeal cancer. Methods:A retrospective analysis was conducted on the clinical data of 91 patients with T3 glottic laryngeal cancer. Results:Among the 91 patients, 58 cases (63.7%) had anterior invasion and 33 cases (36.3%) had posterior invasion. The posterior invasion was significantly correlated with invasions of the dorsal plate of cricoid cartilage (P<0.001), arytenoid cartilage (P= 0.001), and subglottic region(P = 0.001). There was no statistical difference in survival outcomes between the total laryngectomy group and the partial laryngectomy group. But in the partial laryngectomy group, the 5-year disease-free survival(DFS) of patients with anterior invasive tumors was better than that of patients with posterior invasion tumors (HR: 4.681, 95%CI 1.337-16.393, P=0.016), and subglottic invasion was associated with worse loco-regional recurrence-free survival(LRRFS)(HR: 3.931, 95%CI 1.054-14.658, P=0.041). At the same time, we found that involvement of the dorsal plate of cricoid cartilage was an independent risk factor for postoperative laryngeal stenosis in partial laryngectomy patients (HR:11.67, 95%CI 1.89-71.98,P=0.008). Conclusion:Compared with total laryngectomy, selected partial laryngectomy can also achieve favorable oncological outcomes. Posterior invasion and subglottic extension are independent prognostic factors for recurrence of partial laryngectomy in T3 glottic laryngeal cancer, and the involvement of the dorsal plate of cricoid cartilage is associated with postoperative laryngeal stenosis. The tumor invasion pattern of laryngeal cancer should be further subdivided in order to select a more individualized treatment plan.
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Humanos , Prognóstico , Neoplasias Laríngeas/patologia , Estudos Retrospectivos , Laringoestenose/cirurgia , Carcinoma de Células Escamosas/patologia , Complicações Pós-Operatórias/cirurgia , LaringectomiaRESUMO
Objective:To investigate the diversity and clinical effect of supraclavicular island flap in repairing the defect after head and neck tumor surgery. Methods:A retrospective analysis was performed on 30 patients who received the repair of head and neck defects with supraclavicular island flaps at Department of Otorhinolaryngology Head and Neck Surgery of the First Affiliated Hospital of Chongqing Medical University from January 2017 to March 2023. The sites and types of defects, intraoperative blood loss, time of flaps preparation, areas of flaps, survival of the flaps and other complications were recorded. Results:A total of 30 patients were enrolled, including 26 males and 4 females, aged 36-82 years. Among them, 22 patients with hypopharyngeal partial defect were repaired (19 patients with ipsilateral defect and 3 patients with contralateral defect). In addition, 2 patients were repaired with contralateral pectoralis major musculocutaneous flap around the hypopharynx, the neck skin defect was repaired in 2 patients, the parotid skin defect was repaired in 2 patients, the temporal bone skin defect was repaired in 1 patient, and the cervical esophageal defect was repaired in 1 patient. The average blood loss during the operation was 8 ml, and the average time was 32 min. The flap areas ranged from 5.0 cm×4.0 cm to 20.0 cm×8.0 cm. 27 of 30 flaps survived(90.0%), and pharyngeal fistula occurred in 6 patients after operation(4 flaps survived after local dressing). One patient was complicated with venous thrombosis(the flap necrosis after local dressing). Shoulder and neck functions(lift, internal rotation and abduction) were not significantly affected in 29 patients, and the function of 1 patient with shoulder infection was not affected after treatment. Conclusion:Supraclavicular island flap is a highly vascularized axial fascial flap. It is easy to make, thin, and soft in texture, and can be used to repair different sites and types of postoperative head and neck tumor defects with a low donor site complication rate. Good results in post-operative repair of head and neck tumors are worth promoting.
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Masculino , Feminino , Humanos , Procedimentos de Cirurgia Plástica , Estudos Retrospectivos , Transplante de Pele , Lesões dos Tecidos Moles/cirurgia , Resultado do Tratamento , Retalhos Cirúrgicos , Neoplasias de Cabeça e Pescoço/cirurgiaRESUMO
Objective: To establish a method for the induction of peripheral blood mononuclear cells to hepatocyte-like cells, and preliminarily investigate cell response to injury under the effect of acetaminophen (APAP). Methods: The surface marker CD45 of peripheral blood mononuclear cells wase detected cells by using flow cytometry and immunofluorescence methods. The cellular morphology of induced hepatocyte-like cells was observed under an inverted microscope. Real-time fluorescent quantitative PCR (RT-PCR) was used to detect the expression level of hepatocyte-specific genes, such as cytochrome (CY) P1A2, CYP3A4, CYP2C9, albumin (ALB), alpha-fetoprotein (AFP), and hepatocyte nuclear factor (HNF)4α mRNA. Immunofluorescence method was used to detect intracellular hepatocyte markers AFP, HNF4α, and ALB expression at the protein level. Biochemical analyzer was used to detect hepatocyte-specific secretory functions of AFP, ALB, and urea. Luciferase chemiluminescence method was used to detect the activity of key drug metabolizing enzyme CYP3A4. Colorimetric assay was used to detect the effect of the drug acetaminophen on hepatocyte-like cells, and alanine aminotransferase (ALT) was used as an indicator of liver cell injury. The statistical differences between the data were compared with t-test and rank-sum test. Results: The positive expression rate of CD45 cell surface markers isolated from peripheral blood mononuclear cells was about 98%, and hepatocyte-like cell morphology changes appeared on 15th day of induction. Compared with isolated mononuclear cells, CYP1A2, CYP3A4, CYP2C9, ALB, AFP and HNF4α mRNA was markedly elevated. The expression level of AFP, ALB and HNF4α protein were equally increased, and the secretory function of AFP, ALB and urea were enhanced. Compared with primary hepatocytes, CYP1A2, CYP2C9, AFP, HNF4α mRNA, and CYP3A4 mRNA did not decrease. The expression levels of AFP, ALB, and HNF4α proteins in the cells did not decrease, and the secretory function of AFP, ALB, and urea did not decrease. In addition, the CYP3A4 enzyme activity produced by hepatocyte-like cells was similar to that of primary hepatocytes. Compared with hepatocyte-like cells incubated without APAP, hepatocyte-like cells incubated with APAP had higher ALT level. Under the effect of APAP, the ALT level of hepatocyte-like cells was higher than isolated mononuclear cells. Conclusion: Peripheral blood mononuclear cells can be induced into hepatocyte-like cells with partial characteristics of hepatocytes, including the activity of CYP3A4, a key enzyme of hepatocyte drug metabolism. Additionally, preliminarily ALT secretory features reflect the hepatocytes injury under the effect of acetaminophen.
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Acetaminofen/farmacologia , Diferenciação Celular , Células Cultivadas , Hepatócitos , Leucócitos Mononucleares , RNA MensageiroRESUMO
OBJECTIVE@#To investigate the clinical effect of titanium cable assisted fixation combined with lengthened Gamma nail in the treatment of subtrochanteric fracture of femur.@*METHODS@#From January 2010 to January 2018, 45 patients with long subtrochanteric fractures were retrospectively analyzed, and they were divided into two groups according to the operation plan. There were 24 cases of titanium cable assisted fixation combined with lengthened Gamma nail (observation group), including 14 males and 10 females, with an age of (54.2±12.8) years; 21 patients (control group) were treated with simple lengthened Gamma nail (control group), including 12 males and 9 females, with an age of (51.4±15.9) years. The operation time, intraoperative blood loss, hospital stay, hemoglobin decrease and fracture healing time were observed and recorded in the two groups. Harris hip function score was used to evaluate the postoperative curative effect, and the occurrence of complications was observed.@*RESULTS@#All patients were followed up for 14 to 36 months. There was significant difference in operation time, intraoperative blood loss, hospital stay between two groups(@*CONCLUSION@#Lengthened Gamma nail fixation is reliable in the treatment of subtrochanteric fracture. Limited incision titanium cable assisted fixation can improve the reduction rate and fixation strength of long segment fracture, which is conducive to early functional exercise.
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Feminino , Humanos , Masculino , Pinos Ortopédicos , Fêmur , Fixação Interna de Fraturas , Fixação Intramedular de Fraturas , Fraturas do Quadril/cirurgia , Estudos Retrospectivos , Titânio , Resultado do TratamentoRESUMO
Nitidine chloride (NC) is a compound with prominent anti-tumor activity. To determine potential cardiotoxicity of NC, this study was designed to investigate the distribution of NC in rat heart and the underlying mechanism. The animal studies were approved by Institutional Animal Care and Use Committee of Zhejiang University Medical Center (2015-380-01) and complied with the standards of animal welfare in China. At 0.25, 0.5 and 2 h after a single intravenous injection (iv) of 5 mg·kg-1 NC, the concentrations of NC in rat heart were 47.7, 71.1 and 63.2 μg·g-1 respectively, which were 576, 1 352 and 1 212 folds of that in plasma. This study also revealed that the NC concentration in heart was 458.5 μg·g-1 (7 336 folds of that in plasma) at 2 h after the last dose in rats, after daily iv administration of NC at 5 mg·kg-1·day-1 for successive 20 days. Further studies showed that the accumulations of NC in MDCK-hOCT1 and MDCK-hOCT3 cells were 16.1 and 4.99 folds higher than that of the mock cells, respectively. There is no significant difference between the accumulations of NC in MDCK cells transfected with hOCTN1, hOCTN2 or hPMAT and the mock cells. Additionally, quinidine, L-tetrahydropalmatine and Decynium 22, the inhibitors of OCTs, clearly reduced the accumulations of NC in primary cardiomyocytes and cardiac fibroblasts from neonatal rats. MTT assay showed that the LC50 of NC on cardiomyocytes and cardiac fibroblasts were 10.9 and 10.4 μmol·L-1, respectively. Moreover, treatment of the primary cardiomyocytes and cardiac fibroblasts with NC (1~15 μmol·L-1) for 48 h resulted in significantly increased LDH enzyme leakage. These results indicated that NC can be highly accumulated in the heart, and accumulation is mediated by OCT1 and OCT3, but not by OCTN1, OCTN2 and PMAT. The accumulated NC has potential cytotoxicity as shown in the results from primary cardiomyocytes and cardiac fibroblasts.
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Antrodia camphorata, a well-known and highly valued edible medicinal mushroom with intriguing activities like liver protection, has been traditionally used for the treatment of alcoholic liver disease. A. camphorata shows highly medicinal and commercial values with the demand far exceeds the available supply. Thus, the petri-dish cultured A. camphorata (PDCA) is expected to develope as a substitute. In this paper, nineteen triterpenes were isolated from PDCA, and thirteen of them were the unique anthroic acids in A. camphorata, including the main content antcin K, which suggested that PDCA produced a large array of the same anthroic acids as the wild one. Furthermore, no obvious acute toxicity was found suggesting the edible safety of PDCA. In mice alcohol-induced liver injury model, triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and malondialdehyde (MDA) had been reduced by the PDCA powder as well as the main content antcin K, which indicated that the PDCA could protect alcoholic liver injury in mice model and antcin K could be the effective component responsible for the hepatoprotective activities of PDCA against alcoholic liver diseases.
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Animais , Feminino , Masculino , Camundongos , Alanina Transaminase , Sangue , Aldeído Desidrogenase , Sangue , Antrodia , Química , Aspartato Aminotransferases , Sangue , Produtos Biológicos , Química , Farmacologia , Usos Terapêuticos , Doença Hepática Induzida por Substâncias e Drogas , Colestenos , Química , Farmacologia , Usos Terapêuticos , VLDL-Colesterol , Sangue , Modelos Animais de Doenças , Etanol , Toxicidade , Carpóforos , Química , Fígado , Metabolismo , Patologia , Hepatopatias Alcoólicas , Malondialdeído , Sangue , Estrutura Molecular , Triglicerídeos , Sangue , Triterpenos , Química , Farmacologia , Usos TerapêuticosRESUMO
Objective@#To investigate the incidence of occult cervical lymph node metastasis and the common neck level of metastases in cN0 laryngocarcinoma, and the relationship between the clinicopathologic features of laryngocarcinoma and cervical lymph node metastasis.@*Methods@#A total of 506 cases with cN0 laryngocarcinoma treated at the First Affiliated Hospital of Chongqing Medical University between March 2011 and March 2018 were enrolled, and their medical records and follow-up data were retrospectively analyzed. Of them, 211 cases of were glottic carcinoma in stage T1 without neck dissection and they were observed by clinical follow-up; other 295 cases, including glottic carcinoma, supraglottic carcinoma and hypopharyngeal carcinoma in stage T2-T4 were treated with surgical resection of the primary lesions and selective neck dissection. SPSS 22.0 software was used to analyze the data.@*Results@#The total incidence of cervical lymph node metastasis was 10.87%(55/506), with a lower incidence in T1 stage glottic carcinoma(6/211,2.84%) than that in other cases(49/295,16.61%). The incidence of cervical lymph node metastasis in glottic carcinoma (29/426, 6.81%) was lower than those in supraglottic carcinoma (22/71,30.99%) and subglottic carcinoma (4/9) (χ2=35.810,P<0.01).The pN+ rates of glottic carcinoma at T1, T2, T3 were 2.84%(6/211), 5.31%(6/113), 16.05%(13/81), and 19.05%(4/21), respectively (χ2=18.572, P<0.01). The pN+ rates of supraglottic carcinoma at T2, T3 and T4 were 3/13, 32.50%(13/40) and 6/13, respectively (χ2=3.649,P>0.05). The incidence of cervical lymph node metastasis in poorly differentiated carcinoma (17/42, 40.48%) was higher than those in moderately differentiated carcinoma (26/205, 12.68%) and high differentiated carcinoma(12/246, 4.88%)(χ2=36.356, P<0.01). Moreover, 85 pN+ lymph nodes were obtained by selective neck dissection, respectively 43(50.59%) in level Ⅱa, 30(35.29%) in level Ⅲ, 1(1.18%) in level Ⅳ and 11(12.94%) in level Ⅵ.@*Conclusions@#The occult cervical lymph node metastasis was frequently found in cN0 laryngocarcinoma. Selective neck dissection should be performed with surgery for the primary lesions in T3-T4 glottic laryngeal cancer, T2-T4 supraglottic laryngeal cancer and subglottic carcinoma, and the neck dissection for level Ⅱa and Ⅲ is appropriate. It is required to detect pre-laryngeal and pre-tracheal lymph nodes in patients with subglottic laryngeal carcinoma.
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Multidisciplinary team is the basic model of modern head and neck oncology treatment. We try to integrated multimedia presentation in multidisciplinary head and neck oncology clinical teaching. Through building a multidisciplinary platform for discussion , on the basis of teacher's instruction and student's self-learning, we integrate the related knowledge with multimedia presentation and computer net-work to guide the learning of core knowledge and key problems of head and neck cancer, thus forming a new clinical teaching mode combining theory study with practice training. In the process of implementation, we should pay attention to arranging the learning tasks in advance, and highlight the key learning content in the multidisciplinary discussion, so as to improve the students' interest and enthusiasm in learning.
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Objective To evaluate the cytotoxicity of natural killer(NK)-92 cell lines against various human hepatocellular carcinoma cells(HCCs)and to explore the potential mechanism.Methods We established a culture method of NK-92 cell lines in vitro.Lactate debydrogenase(LDH)cytotoxicity assays and cytokine release assays were performed to determine whether NK-92 cell lines could recognize and kill HCCs in vitro.At the same time,Nu/Nu mices were housed. Subcutaneous(sc)xenografts HepG2 models of human hepatocellular carcinoma were established.1×107NK-92 cells were intravenously(iv)injected through the tail vein on days 2,9,16,23 while the control group was injected with PBS in the same way.Tumor size, tumor volume, tumor mass and mouse survival status were closely observed in experimental and control groups.Mice were euthanized when tumor-bearing time reached 28 days.Xenograft tissues were taken for general observation.Sections were cut and processed for HE staining and immunofluorescence staining.The expression of glypican-3(GPC3)protein in xenografts tissue was clearly defined.Results NK-92 cell lines that were chronically cultured in vitro and maintained typical phenotypic characteristics of NK cells with good cellular activity.Enhanced cytotoxicity and IFN-γ production of NK-92 cell lines were identified by LDH and ELISA,indicating that NK-92 cell lines could recognize and kill different kinds of HCCs.In addition,NK-92 cell lines efficiently suppressed the growth of HCC xenografts in vivo.Tumor volume in experimental group was significantly reduced compared with control group and there was low a GPC 3 expression in experimental group through immunofluorescence and immunohistochemistry results, pointing to the possibility that the cytotoxicity of NK cells was correlated with GPC3 +HCCs.Conclusion NK cells provide a promising means of therapeutic intervention for HCCs.NK-92 cell lines could eliminate HCC cells in vitro and in vivo.The cytotoxicity of NK-92 cell lines may work by killing the GPC3-positive cells in the liver cancer tissue.In addition to the anti-tumor effect, NK cells also have cytotoxicity on pathogens such as bacteria and viruses.
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Objective To generate hemogenic endothelial cells(HECs)from human induced pluripotent stem cells (hiPSCs)in vitro in order to learn more about the mechanism by which the vascular niche affects HECs production and self -renewal.Methods hiPSCs with reporter gene runx1c were differentiated to hematopoietic cells by spinEB method.The CD34 positive cells were sorted by magnetic-activated cell sorting(MACS)at day 10 after hematopoietic differentiation. Afterwards,these CD34 positive cells were co-cultured with DLL4 overexpressed vascular niche cells VeraVec to further differentiate to HECs.The HECs derived from the hiPSCs were characterized by FACS.Results We first established an hiPSCs single cell culture method for spinEB differentiation.Single cell cultured hiPSCs with reporter gene runx 1c were differentiated to form embryonic bodies(EBs)by spinEB method.The HECs were enriched from the day 10.Meanwhile, we cultured the E4ORF1 transfected human umbilical vein endothelial cell(HUVEC)line(VeraVec)and examined the expression of NOTCH signaling pathway related genes.According to the results, VeraVec had a high expression level of NOTCH ligand DLL4 at both mRNA and protein levels.And the CD34 positive HECs were co-cultured with DLL4 overexpressed VeraVec cells,which promoted the expression of tdTomato during hematopoitic differentiation and increased HSCs production.Conclusion A method of inducing hiPSCs differentiation by spinEB has been established, which can enrich HECs.This model can be applied to study the mechanism by which the vascular niche promotes hematopoietic differentiation from hPSCs.The generated functional HSCs are of great social and military values for HSCs transplantation and battlefield radiation injury treatment.
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<p><b>OBJECTIVE</b>To study the effect of diallyl thiosulfinate (DATS) on the proliferation of side population (SP) cells in multiple myeloma (MM) and its mechanism.</p><p><b>METHODS</b>RPMI-8226 and NCI-H929 cells were cultured, and the level of SP cells was detected by Hoechst33342 staining. The SP cells were cultured and treated with 10 µg/ml DATS, the CCK8 assay was carried out to examine the effect of DATS on the proliferation ability in SP cells, and plate colony-forming test was used to examine the colony-forming ability, the flow cytometry assay was carried out to examine the cell cycle, Western blot assay was used to examine the expression of cyclin D1, cyclin E, CDK2 and CDK4.</p><p><b>RESULTS</b>SP cells were detected in RPMI-8226 and NCI-H929 cells with a proportion of 3.17±0.98 and 2.65±0.61, respectively. DATS treatment could significantly inhibit the SP cells survival in a time-dependent manner, and also could significantly inhibit the colony forming. In addition, DATS treatment could significantly induce the G/S arrest and suppress the expression of cyclin D1, cyclin E, CDK2 and CDK4.</p><p><b>CONCLUSION</b>DATS can inhibit the proliferation and colony-forming of SP cells in multiple myeloma, and induce the G/S arrest that may be carried out via suppressing the expression of cyclin D1, cyclin E, CDK2 and CDK4.</p>
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This study aims to explore the differences of event-related potential (ERP) between attention deficit hyperactivity disorder (ADHD) and normal children, so that these differences provide scientific basis for the diagnosis of ADHD. Eight children were identified to be ADHD group by the diagnostic criteria of DSM-IV (diagnostic and statistical manual of mental disorders-IV), and the control group also consisted of 8 normal children. Modified visual-continuous performance test (CPT) was used as the experiment paradigm. The experiment included two major conditions, i. e. Go and NoGo. All the 16 subjects participated in the study. A high density EEG acquisition instrument was used to record the EEG signal and processed these EEG data by means of ERP and spectrum analysis. P2-N2 peak-peak value and spectral peak around 11 Hz were analyzed between ADHD subjects and those in the control group, and then statistical tests were applied to these two groups. Results showed that: (1) Under the condition of Go, ADHD group had a significant lower P2-N2 peak-peak value than the values in the control group (P < 0.05); but under the condition of NoGo there was no significant difference in between. (2) Compared with the control group, the ADHD group had significant lower spectral amplitude around 11 Hz under the condition of NoGo (P < 0.05). However, under the condition of Go the difference was insignificant. In conclusion, there is certain cognitive dysfunction in ADHD children. P2-N2 peak-peak value and spectral peak around 11 Hz could be considered as clinical evaluation indexes of ADHD children's cognitive function. These two objective indexes provide an early diagnosis and effective treatment of ADHD.
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Criança , Humanos , Transtorno do Deficit de Atenção com Hiperatividade , Diagnóstico , Encéfalo , Estudos de Casos e Controles , Transtornos Cognitivos , Diagnóstico , Eletroencefalografia , Potenciais EvocadosRESUMO
This article studied the chemical constituents from the aerial part of Vitis thunbergii var. taiwaniana. The 60% ethanol extract was eluted with 95% ethanol though HP-20 macroporous adsorption resin column. 12 compounds, including (1) betulinic acid, (2)2, 2, 2'-bis (4-hydroxyphenyl) propane bis (2, 3-epoxypropyl) ether, (3) eriodictyol, (4) trans-ε-viniferin, (5) (+)-cis-ε-viniferin, (6) kobophenol A, (7) ampelopsin A, (8) nepalensinol B, (9) cis-miyabenol C, (10) cis-vitisin B, (11) cis-gnetin H and (12) (+)-hopeaphenol, were separated by using normal phase silica gel, ODS, Sephdadex LH-20 column chromatographies and semi-preparative or preparative HPLC. Compounds 2, 5, 6, 8, 9, 10, 11 were separated from the genus Vitis for the first time and compounds 3, 7, 12 were separated from Vitis thunbergii var. taiwaniana for the first time. At a concentration of 50 μmol · L(-1), compound 6, 7 and 11 showed strong cytotoxicity against MCF-7 cell lines with the inhibition rate of 66.58%, 57.16%, 52.84%, respectively.
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Humanos , Antineoplásicos Fitogênicos , Farmacologia , Células MCF-7 , Extratos Vegetais , Farmacologia , Vitis , QuímicaRESUMO
OBJECTIVE@#To investigate whether serum thyroid-stimulating hormone(TSH) levels are of value in predicting malignancy in patients with nodular thyroid disease. The relationship between TSH levels and thyroid cancer incidence was also investigated.@*METHOD@#One hundred and eight patients with thyroid tumors who underwent surgical treatment were included in this study (25 cases of papillary thyroid cancer and 83 cases of benign tumors). The data of their preoperative serum TSH level, gender, age, number of tumors detected by ultrasonic inspection, and pathological type were retrospectively analyzed, and their association with thyroid cancer incidence was explored. Logistic regression analysis was used to predict thyroid cancer risk factors.@*RESULT@#Patients with malignancy had a higher mean value of TSH than that of the patients with benignancy [(1.94±1.01)mlU/L vs (1.16± 0.85)mIU/L, respectively; P<0.05]. Compared with the patients below the population mean, patients above the population mean had a significantly higher malignancy rate (35.9% vs 15.9%, P<0.05). High serum TSH level (OR=10.913, P=0.001), male (OR=4. 845, P=0.028) and age ≥45 (OR=10.831, P=0.001) are independent risk factors of thyroid cancer.@*CONCLUSION@#The preoperative serum TSH level may be useful in predicting the probability of cancer. The high serum TSH level, male, age ≥45 are risk factors of thyroid cancer.
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Humanos , Masculino , Carcinoma , Patologia , Carcinoma Papilar , Incidência , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide , Patologia , Nódulo da Glândula Tireoide , Sangue , Tireotropina , SangueRESUMO
OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.
RESUMO
<p><b>OBJECTIVE</b>To study the chemical constituents of Elephantopus tomentosus.</p><p><b>METHOD</b>The compounds were isolated by repeated HP20 macro porous adsorption resin column combined with Sephadex LH-20, ODS and silica gel chromatographies. The structures were identified on the basis of extensive spectroscopic data analysis and by comparison of their spectral data reported.</p><p><b>RESULT</b>Eighteen compounds were identified as 2-deethoxy-2beta-hydroxyphantomolin (1), 2beta-hydroxy-2-deethoxy-8-O-deacylphantomolin-8-O-tiglinate (2), 2beta-methoxy-2-deethoxyphantomolin (3), 2beta-methoxy-2-deethoxy-8-O-deacylphantomolin-8-O-tiglinate (4), molephantin (5), molephantinin (6), tricin (7), luteolin (8), quercetin (9), 3beta-friedelinol (10), 3beta-hydroxyolean-12-en-28-oic acid (11), 3, 5-di-O-caffeoyl quinic acid (12), 3,4-di-O-caffeoyl quinic acid (13), syringaresinol-4-beta-D-glucopyranoside (14), xylogranatinin (15), byzantionoside B (16), 2'-hydroxycinnamaldehyde (17), and caffeic acid ethyl ester (18).</p><p><b>CONCLUSION</b>Compounds 9, 11, 14-18 were separated from Elephantopus for the first time.</p>