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This study aimed to investigate the effects of Erxian Decoction(EXD)-containing serum on the proliferation and osteogenic differentiation of MC3T3-E1 cells under oxidative stress through BK channels. The oxidative stress model was induced in MC3T3-E1 cells by H_2O_2, and 3 mmol·L~(-1) tetraethylammonium(TEA) chloride was used to block the BK channels in MC3T3-E1 cells. MC3T3-E1 cells were divided into a control group, a model group, an EXD group, a TEA group, and a TEA+EXD group. After MC3T3-E1 cells were treated with corresponding drugs for 2 days, 700 μmol·L~(-1) H_2O_2 was added for treatment for another 2 hours. CCK-8 assay was used to detect cell proliferation activity. The alkaline phosphatase(ALP) assay kit was used to detect the ALP activity of cells. Western blot and real-time fluorescence-based quantitative PCR(RT-qPCR) were used to detect protein and mRNA expression, respectively. Alizarin red staining was used to detect the mineralization area of osteoblasts. The results showed that compared with the control group, the model group showed significantly blunted cell proliferation activity and ALP activity, reduced expression of BK channel α subunit(BKα), collagen Ⅰ(COL1), bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG), and phosphorylated Akt, decreased mRNA expression levels of Runt-related transcription factor 2(RUNX2), BMP2, and OPG, and declining area of calcium nodules. EXD-containing serum could significantly potentiate the cell proliferation activity and ALP activity, up-regulate the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt, and forkhead box protein O1(FoxO1), promote the mRNA expression of RUNX2, BMP2, and OPG, and enlarge the area of calcium nodules. However, BK channel blockage by TEA reversed the effects of EXD-containing serum in promoting the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt and FoxO1, increasing the mRNA expression of RUNX2, BMP2, and OPG, and enlarging the area of calcium nodules. EXD-containing serum could improve the proliferation activity, osteogenic differentiation, and mineralization ability of MC3T3-E1 cells under oxidative stress, which might be related to the regulation of BK channels and downstream Akt/FoxO1 signaling pathway.
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Osteogênese , Subunidade alfa 1 de Fator de Ligação ao Core/farmacologia , Canais de Potássio Ativados por Cálcio de Condutância Alta/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Cálcio/metabolismo , Diferenciação Celular , RNA Mensageiro/metabolismo , Proliferação de Células , OsteoblastosRESUMO
To establish the quantitative analysis multi-components with a single-marker(QAMS) method for six components and fingerprint of standard decoction of Gastrodiae Rhizoma, verify the accuracy and feasibility of the method, and evaluate the quality of standard decoction. Based on UPLC with gastrodin as the internal standard, relative correction factors of p-hydroxybenzyl alcohol, parishin E, parishin B, parishin C, parishin A and gastrodin were determined by investigating the column temperature, flow rate, chromatographic columns and multi-point concentration correction. The total contents in 18 batches of standard decoction of Gastrodiae Rhizoma and the similarity were determined to calculate the similarity. The results of standard curve method, external standard one-point method and quantitative analysis multi-components with a single-marker(QAMS) were compared, and the results showed that there was no significant difference among these three methods. By analyzing the results of standard decoctions from different origins, it can be seen that the quality of Gastrodia standard decoctions derived from Anhui and Yunnan was better, followed by Shaanxi and Hubei, and relatively poor in Gansu, with similarities all above 0.90 in the fingerprints. Therefore, the QAMS method that can measure the contents of gastrodin, p-hydroxybenzyl alcohol, parishin E, parishin B, parishin C and parishin A in standard decoction of Gastrodiae Rhizoma combined with fingerprint is accurate, feasible and fast, which can be used to evaluate the quality of standard decoction of Gastrodiae Rhizoma, and also provide a reference for the research on the quality standards of raw materials for Gastrodiae Rhizoma prepared slices and alike.
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Medicamentos de Ervas Chinesas , Gastrodia , China , Cromatografia Líquida de Alta Pressão , Padrões de Referência , RizomaRESUMO
Flood disaster is one of the most serious natural disasters in the world, and it could pose an inestimable impact on the affected people. Based on existing laws, regulations, and emergency manuals in China, extensive literature review, epidemiological and related protection evidence, and expert consultation, this study analyzed different health risk factors of flood disaster and proposed a multi-stage, multi-population, and multi-phase comprehensive protection measures for the public in the perspective of pre-event prevention, in-event intervention and post-event rescue strategy, which could provide a scientific basis for improving the level of public health protection against the flood disaster and corresponding health outcomes.
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BACKGROUND: Xianling Gubao Capsule can reduce bone loss and promote the formation of osteoblasts. Its efficacy in osteoporosis and fracture has been preliminarily confirmed, but its efficacy in postmenopausal female periodontitis has not been clarified. OBJECTIVE: To assess the effect of Xianling Gubao Capsule on alveolar bone mass in postmenopausal women with periodontitis by cone-beam CT, providing an effective basis for its clinical use. METHODS: One hundred and thirty-five patients with periodontitis diagnosed and treated in the Department of Stomatology, First Affiliated Hospital of Guiyang College of Traditional Chinese Medicine from October 2015 to April 2017 were enrolled. All patents were randomly divided into blank group (basic treatment), control group (basic treatment+vitamin D) and experimental group (basic treatment+Xianling Gubao Capsule), with 45 patients in each group. All patients received oral hygiene and health education, and were guided to brush their teeth correctly and self-control plaques. Basic treatments for periodontal disease were performed to eliminate plaque and calculus by cleaning and curing the gums, and smoothing the root surface, once 3 months for 6 continuous months. The control group received oral Calci-D (600 mg of calcium and 125 U of vitamin D per tablet), one tablet per day, for 6 continuous months. The experimental group was orally given Xianling Gubao Capsule 1.0 g/time, 3 times per day, 3 months as a course of treatment, for two consecutive courses. Cone-beam CT was used to detect the changes of bone mineral density and height of alveolar bone defect in all patients before treatment, 3 and 6 months after treatment. Ethics approval was obtained from the First Affiliated Hospital of Guiyang College of Traditional Chinese Medicine on May 29, 2015 (approval No. K2015-186). RESULTS AND CONCLUSION: (1) Compared with the baseline, the alveolar bone defect height was significantly reduced in the experimental and control groups (P 0.05). The improvement of alveolar bone defect height and bone density was better in the experimental group than the control group (P 0.05). The alveolar bone defect height and bone mineral density of the patients older than 55 years had no changes (P > 0.05). (3) Alveolar bone defect height and bone mineral density were significantly improved in patients with mild and moderate periodontitis (P 0.05). To conclude, the treatment of periodontitis by Xianling Gubao Capsule can significantly improve the alveolar bone defect height and bone mineral density, and cone-beam CT can be used to evaluate its clinical efficacy.
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The technical review for clinical trials of implantable sacral nerve stimulation system needs to focus on the basic principles of trials, clinical institutition and design elements of clinical trial protocols.
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BACKGROUND: Adiponectin can regulate glucose and lipid metabolism in the body and protect body tissues and organs. Therefore, we combine it with stem cell transplantation for the treatment of nephrotic syndrome. OBJECTIVE: To investigate the therapeutic effect of adiponectin combined with adipose stem cell transplantation in the rats with nephrotic syndrome. METHODS:Rat adipose stem cells were resuscitated and cultured in vitro,and labeled using CD-Dil prior to the transplantation.Eighty Sprague-Dawley rats were randomly assigned into a normal control group, a model group, an adipose stem cell transplantation group, and a combined group (adiponectin and adipose stem cell transplantation), with 20 rats in each group. Rats in the normal control group received no treatment. The remaining rats in the latter three groups were given a single intravenous injection of adriamycin (6 mg/kg) to prepare animal models of nephrotic syndrome, followed by a tail vein injection of normal saline, 2×106/L adipose stem cell suspension, and 1 μg/kg adiponectin plus 2×106/L adipose stem cell suspension, respectively. Administration of adiponectin in the combined group was given once a day, for 3 consecutive days. RESULTS AND CONCLUSION: (1) Compared with the normal control group, higher levels of 24-hour urinary protein, blood cholesterol and urea nitrogen as well as lower serum albumin level were observed in the remaining three groups at 28 days after modeling. Compared with the model group, the levels of 24-hour urinary protein, blood cholesterol and urea nitrogen in the adipose stem cell transplantation group were decreased, while the level of serum albumin was increased. Compared with the adipose stem cell transplantation group, the levels of 24-hour urinary protein, blood cholesterol and urea nitrogen in the combined group were decreased, while the level of serum albumin was increased. (2) At 28 days after modeling, severest renal damage appeared in the model group, while the damage was reduced in the adipose stem cell transplantation group and considerably relieved in the combined group accompanied by improved inflammation and edema. (3) The number of CM-Dil positive cells in renal tissues was highest in the combined group followed by the adipose stem cell transplantation group, but no positive cells were detected in the normal control and model groups. Our experimental findings reveal that adipose-derived stem cell transplantation has a certain therapeutic effect on nephrotic syndrome, and adiponectin can enhance this therapeutic effect, by improving renal function and effectively inhibiting the pathological changes of renal tissues.
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PURPOSE: This study was designed to investigate the relationship between molecular subtype and locoregional recurrence (LRR) in patients with early-stage breast cancer with 1-3 positive axillary lymph nodes (ALNs) and improve the individualized indications for postmastectomy radiotherapy (PMRT). METHODS: The records of 701 patients with pT1-2N1M0 breast cancer who did not undergo PMRT were retrospectively analyzed. Tumors were subclassified as follows: luminal A, luminal B, human epidermal growth factor receptor 2 (HER2)-enriched, and basal-like subtypes. Multivariate Cox analysis was used to determine the risk of LRR associated with the different subtypes and to adjust for clinicopathologic factors. RESULTS: Luminal A, luminal B, HER2-enriched, and basal-like subtypes accounted for 51.2%, 28.0%, 8.1%, and 12.7% of cases, respectively. The median follow-up duration was 67 months (range, 9-156 months). Univariate analysis revealed that, compared with the luminal A subtype, the HER2-enriched and basal-like subtypes were associated with significantly higher 5-year LRR rates (5.6% vs. 21.6% and vs.15.7% respectively; p=0.002 each), lower 5-year LRR-free survival (LRFS) rates (90.6% vs. 73.8% and 78.5%, respectively; p=0.001 each), and poorer 5-year breast cancer-specific survival (BCSS) rates (93.7% vs. 82.2% [p=0.002] and 84.9% [p=0.001], respectively). Multivariate analysis revealed that the HER2-enriched and basal-like subtypes, age ≤35 years, a medial tumor, and pT2 stage were poor prognostic factors for LRR and LRFS; furthermore, 2 to 3 positive ALNs represented an independent prognostic factor affecting LRR. The 10-year LRR rates of patients with 0, 1, 2, 3, and 4 risk factors were 1.0%, 6.9%, 14.3%, 30.4%, and 54.3%, respectively (p<0.001); the 10-year BCSS rates were 86.6%, 88.5%, 84.4%, 79.7%, and 38.8%, respectively (p<0.001). CONCLUSION: Molecular subtyping allows for individualized evaluation of LRR risk in patients with pT1-2N1M0 breast cancer. PMRT should be recommended for patients with ≥3 LRR risk factors.
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Humanos , Neoplasias da Mama , Mama , Seguimentos , Linfonodos , Tipagem Molecular , Análise Multivariada , Recidiva Local de Neoplasia , Fenobarbital , Prognóstico , Radioterapia , Receptores ErbB , Recidiva , Estudos Retrospectivos , Fatores de RiscoRESUMO
Nature Calculus Bovis is one of the precious traditional Chinese medicines, which is the dried gallstone in gallbladder, bile duct or hepatic duct. In order to meet the clinical needs, we have developed artificial Calculus Bovis, invitro and other varieties. Calculus Bovis mainly contains bile pigments, bile acids, amino acids, inorganic elements and other types of components. Besides bile pigments, bile acids is the second largest of the active ingredients of Calculus Bovis, which is closely related to the antipyretic and analgesic, antiinflammatory effect and depressurization. The article outlines and summarizes the study progress in pharmacological effects and quantitative methods for components of bile acids in Calculus Bovis and its substitutes and preparations, aiming to provide a reference for subsequent research as well as quality control.
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<p><b>BACKGROUND</b>H3N2 subtype influenza A viruses have been identified in humans worldwide, raising concerns about their pandemic potential and prompting the development of candidate vaccines to protect humans against this subtype of influenza A virus. The aim of this study was to establish a system for rescuing of a cold-adapted high-yielding H3N2 subtype human influenza virus by reverse genetics.</p><p><b>METHODS</b>In order to generate better and safer vaccine candidate viruses, a cold-adapted high yielding reassortant H3N2 influenza A virus was genetically constructed by reverse genetics and was designated as rgAA-H3N2. The rgAA-H3N2 virus contained HA and NA genes from an epidemic strain A/Wisconsin/67/2005 (H3N2) in a background of internal genes derived from the master donor viruses (MDV), cold-adapted (ca), temperature sensitive (ts), live attenuated influenza virus strain A/Ann Arbor/6/60 (MDV-A).</p><p><b>RESULTS</b>In this presentation, the virus HA titer of rgAA-H3N2 in the allantoic fluid from infected embryonated eggs was as high as 1:1024. A fluorescent focus assay (FFU) was performed 24-36 hours post-infection using a specific antibody and bright staining was used for determining the virus titer. The allantoic fluid containing the recovered influenza virus was analyzed in a hemagglutination inhibition (HI) test and the specific inhibition was found.</p><p><b>CONCLUSION</b>The results mentioned above demonstrated that cold-adapted, attenuated reassortant H3N2 subtype influenza A virus was successfully generated, which laid a good foundation for the further related research.</p>
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Animais , Cães , Camundongos , Células COS , Chlorocebus aethiops , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Genética , Vírus da Influenza A Subtipo H3N2 , Alergia e Imunologia , Vacinas contra Influenza , Alergia e Imunologia , Camundongos Endogâmicos BALB C , Neuraminidase , Genética , Plasmídeos , Vírus Reordenados , Alergia e Imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vacinas Atenuadas , Alergia e Imunologia , Proteínas Virais , GenéticaRESUMO
<p><b>OBJECTIVE</b>To investigate the clinical features, treatment modalities and the prognosis of nasal type NK/T cell lymphoma.</p><p><b>METHODS</b>The data of 39 such patients treated from June 2000 to December 2003 were retrospectively reviewed. Twenty three patients were treated by combined chemoradiotherapy, basing on anthracycline-containing CHOP or similar regimens (median 5 cycles). Eleven patients by chemotherapy alone, 2 by radiotherapy alone and 2 aged patients by palliative chemotherapy or radiotherapy. Radiotherapy was given by high energy photon ray combined with electron beam with a median curative dose of 56 Gy in conventional fractionation. Bivariate correlations and univariate prognostic factors were analyzed.</p><p><b>RESULTS</b>Median follow-up time for the 21 patients who were still alive was 22.5 months. The overall remission rate (RR) after initial treatment was 66.7% (21 CR, 3 PR). Chemotherapy alone got a CR rate of only 37.5%. The overall local control rate was 59.4%. Local relapse rate after curative radiotherapy was 25.0%. Radiotherapy was positively correlated with local control (P = 0.000) and time to disease progression (TTP, P = 0.002). Skin and intestine were among the extranodal relapse sites. Fifteen patients had highly aggressive tumors with a median survival time of only 5 months. Univariate analysis showed that significant favorable survival prognostic factors were: radiotherapy (P = 0.001); lower risk International Prognostic Index (IPI, P = 0.001); complete remission after primary treatment (P = 0.000); pre-diagnostic history > 2 months (P = 0.024); and free of skin involvement (P = 0.034).</p><p><b>CONCLUSION</b>Most of nasal type NK/T cell lymphoma are in early stage when diagnosed. Radiotherapy remains to be the mainstay of treatment. Combined chemoradiotherapy needs further improvement for the progressive disease type. Some patients may have highly aggressive tumors with poor prognosis. Optimal prognostic factors and individualized treatment regimens need to be investigated.</p>
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Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Combinada , Seguimentos , Células Matadoras Naturais , Linfoma de Células T , Patologia , Terapêutica , Recidiva Local de Neoplasia , Neoplasias Nasais , Patologia , Terapêutica , Prognóstico , Indução de Remissão , Estudos RetrospectivosRESUMO
<p><b>OBJECTIVE</b>To retrospectively analyze the treatment outcomes and prognostic factors of nasal and nasopharyngeal peripheral T cell lymphomas (PTCL) patients.</p><p><b>METHODS</b>One hundred and twelve patients with pathologically confirmed nasal and nasopharyngeal PTCL were included, among which 39 were CD56(+) NK/T cell lymphomas. The median pre-treatment disease course was 4 months. 84 were males and 28 females median age was 46 years. The tumors mainly involved nasal cavity (88 cases) and/or nasopharynx (50 cases) and adjacent structures, and 83 cases with extra-cavity diseases. 91.1% of the patients had Ann Arbor I(E)/II(E) diseases. The International Prognostic Indices (IPI) were less than 2 scores in 78.8% of the patients. Seventy two patients received combined chemo-radiotherapy, 32 chemotherapy only, 3 radiotherapy only and 5 no any treatment.</p><p><b>RESULTS</b>Median follow-up duration was 42 months. Chemotherapy achieved a complete remission (CR) rate of 34.4% for initial treatment, and of 65.1% after primary treatment. The local tumor controlled rate was 50.5%, and the median time to tumor progression (TTP) was 11 months. There were evidences of systemic relapse in more than 30% of the patients. The extra-cavity tumors usually had a shorter TTP (r(s) = -0.191, P = 0.024). The progress-free survival and overall survival rates were 38.8% and 52.4% at 3 years, and 34.9% and 44.8% at 5 years respectively. Univariate analysis showed that favorable prognostic factors for survival were pre-treatment course > 3 months, earlier clinical stage, non NK/T lymphoma, no skin involvement, lower IPI, CR after initial chemotherapy, radiotherapy, CR after primary treatment and local tumor controlled. Multivariate analysis showed that, pre-treatment course > 3 months (P = 0.011), non NK/T lymphoma (P = 0.007), CR after initial chemotherapy (P = 0.008) and radiotherapy (P = 0.000) were favorable prognostic factors for survival.</p><p><b>CONCLUSIONS</b>Although most nasal and nasopharyngeal peripheral T-cell lymphomas were diagnosed at early stage diseases, some of them were highly aggressive with poor prognosis, particularly CD56(+) NK/T cell lymphomas. Combination chemo/radiotherapy, though remained principal treatments, more effective therapeutic modalities are expected.</p>
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Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Terapia Combinada , Tratamento Farmacológico , Métodos , Seguimentos , Estimativa de Kaplan-Meier , Linfoma de Células T Periférico , Patologia , Terapêutica , Neoplasias Nasofaríngeas , Patologia , Terapêutica , Neoplasias Nasais , Patologia , Terapêutica , Prognóstico , Radioterapia , Métodos , Estudos Retrospectivos , Resultado do TratamentoRESUMO
<p><b>OBJECTIVE</b>To study the radiobiological effects of fast neutron/photon mixed irradiation on human cancer cell in vitro and to discuss the mechanism in relation with cell cycle and apoptosis, thus to provide experimental support for the further application of fast neutron radiotherapy of cancer.</p><p><b>METHODS</b>Exponentially growing human nasopharyngeal cancer cell line CNE-1 was irradiated in vitro with 35 MeV p-->Be fast neutron and 6 MV-X ray in grading doses (0 cGy, 40 cGy, 80 cGy, 120 cGy, 160 cGy, 240 cGy, 320 cGy and 400 cGy for neutron, and 0 cGy, 100 cGy, 200 cGy, 300 cGy, 400 cGy, 600 cGy, 800 cGy and 1000 cGy for X ray). Clonogenic assay was performed, and relative biological effectiveness (RBE) of fast neutron was determined with D(10) by means of cell survival curves. Isoeffective doses of 35 MeV p-->Be fast neutron and 6 MV-X ray were obtained according to the RBE. The cells were assigned into two irradiation regimens, (1) the one-week-fractionation regimen, which adopted the radiation pattern of X x 5, N x 2 and X-N-X-X-N. After irradiation the clonogenic assay was performed to compare their survival fractions; (2) the two-dose regimen, with the radiation pattern of X + N, N + X and X + X. Flow cytometry was done at different time points after irradiation to analyze cell cycle distribution and apoptosis. Fast neutron dose was delivered on Tuesday and Friday, and all the other irradiation intervals were 24 h.</p><p><b>RESULTS</b>The RBE of fast neutron to X ray in CNE-1 cells according to the D(10) ratio was 2.40. The neutron isoeffective dose for a single dose of 200 cGy of 6 MV-X ray was approximately 80 cGy. In clonogenic assay, the cell survival fractions were significantly lower in X-N-X-X-N group (0.0079) than those in X x 5 (0.018) and N x 2 (0.017) groups. The flow cytometry suggested a higher percentage of apoptotic cells after mixed irradiation, and different sequence of X ray and neutron irradiations caused varying changes in cell cycle arrest.</p><p><b>CONCLUSION</b>Mixed irradiation of fast neutron and X ray showed a synergic effect in vitro on CNE-1 cell killing. Cell cycle arrest and apoptosis may play some role in the radiation damage repair mechanisms of mixed beam irradiation.</p>
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Humanos , Apoptose , Efeitos da Radiação , Carcinoma de Células Escamosas , Patologia , Ciclo Celular , Efeitos da Radiação , Linhagem Celular Tumoral , Relação Dose-Resposta à Radiação , Nêutrons Rápidos , Usos Terapêuticos , Neoplasias Nasofaríngeas , Patologia , Fótons , Usos TerapêuticosRESUMO
Spider dragline silk is synthesized in special gland named major ampulate (MA) gland. The MA glands were dissected from the abdomen of the spiders Nephila clavata and the total RNA was extracted by the TRIZOL. The cDNA of dragline silk was amplificated by RT-PCR (reverse transcription polymerase chain reaction), multiplex PCR and cloned. PCR identification, restriction analysis and DNA sequence analysis were carried out to verify the recombinant plasmids. The codon usage frequencies of the cloned cDNA were added up, and the predicted amino acid sequence was compared with Spidroin2 of Nephila clavipes. Predicted secondary structure of the predicted amino-acid sequence was analysized by DNAStar software. All results showed that the cloned cDNA we got (GenBank Accession No. AF441245) was the very fragment of spider dragline silk Spidroin2 cDNA.