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Objective To analyze the medication rules of Professor HUANG Feng for the treatment of low back pain using data mining methods.Methods The information of prescriptions for the effective cases of outpatients with low back pain treated by Professor HUANG Feng were collected and screened.Microsoft Excel 2019 was used to analyze the frequency of medication and the distribution of properties,flavors and meridian tropism of the drugs in the included prescription.IBM SPSS Modeler 18.0 was used for association rule analysis,and IBM Statistics 26.0 was used for cluster analysis.Results A total of 239 prescriptions and 75 Chinese medicines were included.There were 23 high-frequency Chinese medicines with the medication frequency being or over 20 times,and the top 10 Chinese medicines were Glycyrrhizae Radix et Rhizoma,vinegar-processed Corydalis Rhizoma,Cibotii Rhizoma,Atractylodis Macrocephalae Rhizoma,Zanthoxyli Radix,salt-processed Achyranthis Bidentatae Radix,Rehmanniae Radix,Dipsaci Radix,Coicis Semen,and Salviae Miltiorrhizae Radix et Rhizoma.The medicines were mainly warm in nature,and were sweet,bitter and pungent in flavor.Most of the drugs had the meridian tropism of liver,stomach and spleen meridians.Among the drug combinations obtained from association rule analysis with the top 20 highest support,vinegar-processed Corydalis Rhizoma,Cibotii Rhizoma,Atractylodis Macrocephalae Rhizoma and Zanthoxyli Radix were the core drugs.Cluster analysis yielded 6 clustering combinations.Conclusion For the treatment of low back pain,Professor HUANG Feng follows the principle of"treatment adapting to the climate,individuality,and environment"and"treating the root cause of the disease",usually adopts the drugs for activating blood,moving qi and relieving pain,nourishing the liver and kidney,and also uses the medicines for replenishing qi and strengthening the spleen.The ideas of HUANG Feng for the treatment of low back pain can be used as a reference for the clinical treatment.
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Objective:To investigate the clinical characteristics, diagnosis and treatment of dermatomyositis with kidney neoplasm.Methods:The data of two patients with dermatomyositis complicated with kidney neoplasm in Tongji Hospital from January to February 2022 were retrospectively analyzed. The first case was a 55-year-old female, who was admitted with the chief complaints of recurrent erythema of upper extremities for 2 months and facial erythema for 1 month. Physical examination: erythema can be seen on upper limbs and face, no tenderness or percussion pain in kidney area. Myositis enzyme profile test showed that anti-Mi-2 antibody and anti-SSA /Ro-52 antibody were positive. Contrast CT showed nodular uneven enhancement in the right kidney with a size of 50 mm×41 mm. The second case was a 58-year-old female, who was admitted with the chief complaints of kidney occupying for a month. Physical examination: flaky erythema on face, no tenderness or percussion pain in kidney area. Myositis enzyme profile test showed that anti-Ro-52 antibody and anti-MDA5 antibody were positive. Contrast CT showed a significantly uneven enhanced mass with a size of about 50 mm×41 mm on left kidney. Both patients were diagnosed with kidney neoplasm before surgery and underwent laparoscopic partial nephrectomy in Tongji Hospital.Results:Both patients received regular oral prednisone after surgery. The pathological presentation of case 1 was papillary renal cell carcinoma, the facial erythema subsided 1 month after surgery, and there was no tumor recurrence for 13 months. The pathological presentation of case 2 was clear cell renal cell carcinoma, facial erythema subsided 2 weeks after surgery, and there was no tumor recurrence for 12 months.Conclusions:The diagnosis of dermatomyositis should be combined with clinical manifestations and laboratory examination, and the possibility of malignant tumor should be excluded due to the high likelihood of concomitant malignancy. For patients with dermatomyositis with kidney neoplasm, the main treatment is still surgery, and supplemented with glucocorticoid therapy.
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Objective:Efficacy and safety of robot-assisted laparoscopic adrenalectomy as a treatment for large adrenal tumors.Three-dimensional(3D) reconstruction can effectively assist in preoperative planning of robotic adrenalectomy and reduce potential complications.Methods:We retrospectively reviewed the relevant information of patients who had a preoperative 3D reconstruction and underwent RA for adrenal masses larger than 10 cm. Thirteen male patients and sixteen female patients were included. The median(range) age was 43(25, 57) years old and the median tumor diameter was 12.1(10.3, 16.2) cm. The patients underwent preoperative CT enhancement scanning, and three-dimensional images were reconstructed based on the examination data. Robot-assisted laparoscopic adrenalectomy was performed under general anesthesia in 29 cases in this cohort.Results:All surgeries were completed successfully without major complications such as massive bleeding, secondary surgery, or even patient death. The median operative time was 131 (80, 245) min, and the median intraoperative bleeding was 330 (50, 2 200 ml) ml. 9 patients received blood transfusions. There were 11 cases of pheochromocytoma (37.9%), 10 cases of adenocarcinoma (34.5%) as well as 2 cases of teratoma (6.9%) and 6 cases of cortical carcinoma (20.7%). The patients were followed up for a median of 30 months after surgery. Except for 3 cases lost to follow-up and 2 patients with cortical cancer who developed recurrence or metastasis after surgery and died at 16 and 23 months after surgery, respectively, the remaining 24 cases have survived to date.Conclusions:RA is a safe and effective treatment for huge adrenal tumors. The 3D reconstruction could help the preoperative planning of RA and reduce potential complications.
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A case of metachronous bilateral renal pelvic carcinoma was reported. A 55-year-old women underwent left nephroureterectomy for the left renal pelvis cancer in 2011, then she was diagnosed with right renal pelvis carcinoma because of intermittent hematuria in 2014.A transurethral ureteroscopic holmium laser resection of the right renal pelvic tumor, partial right pelvis resection and nephrostomy, instillation with hydroxycamptothecin were taken sequentially to delay the dialysis for 53 months. In 2018, the patient underwent right nephroureterectomy because of recurrence of right renal pelvic carcinoma. The patient was followed up for 17 months postoperatively and there was no recurrence. In this case, patient's renal function was protected by the premise tumor control through a variety of minimally invasive and pharmaceutical therapy, which can provide a reference for the kidney-preserving treatment of high-grade renal pelvis cancer.
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OBJECTIVE@#To analyze the genotypes and distribution of thalassemia in children in Quanzhou Region so as to provide reference for the prevention and control of thalassemia.@*METHODS@#A total of 1 302 children with suspected thalassemia were collected from January 2014 to April 2020 in Quanzhou Region. The deletional α-thalassemia was detected by Gap-PCR, and DNA reverse dot blot (RDB) hybridization was used to detect α- and β-thalassemia mutations.@*RESULTS@#In the 1 302 cases, 667 cases were identified as thalassemia carriers, and the positive detection rate was about 51.23%. Among them, 380 cases of α-thalassemia gene were detected, and --@*CONCLUSION@#There are various genotypes of thalassemia in children in Quanzhou Region, and many children with thalassemia major or intermedia. Therefore, further prevention and control of thalassemia need to be strengthened for reducing the birth of thalassemia major or intermedia.
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Criança , Humanos , China , Testes Genéticos , Genótipo , Heterozigoto , Mutação , Talassemia alfa/genética , Talassemia beta/genéticaRESUMO
Objective:To investigate the protective effect of glycyrrhizic acid (GA) from tumor necrosis factor-α+actinomycetes ketone (TNF-α+CHX) induced apoptosis in rat small intestine crypt epithelial cell line (IEC-6) via AU rich element mRNA binding protein HuR mediated posttranscription of p21 and the potential mechanism. Method:The cultured IEC-6 cells were observed. The experiment was divided into blank group, GA (60 μmol·L-1) group, TNF-α+CHX group and GA+TNF-α+CHX group. Cytoplasmic and nuclear HuR were measured by Western blot. The interaction of HuR and p21 mRNA was detected by biotin pull down and RNA IP. Luciferase activity was measured after transfection with construct with p21 3'-UTR cloned into downstream of luciferase reporter. Cell apoptosis was detected by real-time dynamic cell analyzer, p21 and cysteine proteinas-3 precursor protein(proCaspase-3) association was analysised by CO-IP. Result:After GA treatment for 48 h, cytoplasmic HuR protein expression increased(P<0.05),the binding between HuR and p21 mRNA expression up regulated(P<0.05), luciferase activity increased(P<0.01), and p21 mRNA and protein expression also increased(P<0.05), while these results were abolished by HuR silencing with siRNA. GA enhanced p21 and procaspase3 interaction(P<0.05), and attenuated TNF-α+CHX induced apoptosis in IEC-6 cells. Conclusion:GA protected IEC-6 cells from TNF-α/CHX induced apoptosis via HuR mediated p21 posttranscription, which due to GA enhanced HuR binding to endogenous and recombinant p21 mRNA and increased p21 interaction with proCaspase3.
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OBJECTIVE:To establish pre -column derivatization-HPLC fingerprint of Polyporus polysaccharide ,and to determine the contents of main monosaccharide components ,so as to provide reference for quality evaluation of Polyporus umbellatus. METHODS :Polyporus polysaccharide was extracted with boiling water and precipitated by ethanol and deproteinized by Sevage from 11 batches of P. umbellatus from different producing areas. The samples were firstly hydrolyzed with trifluoro-acetic acid (TFA)and then derivatized by 1-phenyl-3-methyl-5-pyrazolone(PMP). HPLC analysis was then conducted. The determination was carried out on HypersiL BDS C 18 column with mobile phase composed of 0.1 mol/L phosphate buffer (pH 6.84)-acetonitrile(84∶16,V/V)by gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm, and column temperature was 30 ℃. The sample size was 20 µL. The similarity of 11 batches of Polyporus polysaccharide was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System (2012A edition ),and the contents of main monosassharide components were detected. The peak was identified by comparing with the reference substance ,and cluster analysis was performed by using SPSS 23.0 software. RESULTS :In HPLC fingerprints of the 11 batches of samples ,3 common peaks were identified ,namely mannose ,glucose and galactose. The similarity of all samples was above 0.94. Cluster analysis classified 11 batches of samples into three categories. S 1-S6,and S 8 were grouped into category 1;S7,S10 and S 11 were grouped into category 2;S9 was individually grouped into one category. Results of content determination showed that the contents of mannose ranged from 1.571 to 8.771 mg/g;those of glucose ranged from 26.072 to 132.194 mg/g,and those of galactose ranged from 3.420 to 36.593 mg/g. CONCLUSIONS :Established pre-column derivatization HPLC fingerprints can provide reference for quality evaluation of P. umbellatus . The monosaccharide composition of different batches of Polyporus polysaccharide is the same ;there is no significant correlation between fingerprint characteristic peak and the origin of herbs ;there is significant difference in the content of monosaccharide of P. umbellatus .
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Objective The article aimed to identify health-related quality of life(HRQOL) themes of patients with Crohn's disease(CD) and establish a HRQOL model by a meta synthesis of qualitative studies on CD patients′ HRQOL. Methods A retrieval of HRQOL-related qualitative studies on CD patients was conducted in databases including DirectPsycINFO, VIP, etc and the results were integrated by integrating method. Results A total of 8 researches were included to refine 44 results which were integrated into 15 themes. The themes were further grouped into 6 HRQOL domains: physical function, psychological function, social function, study and work skills, sexual function, perception of health and well-being. Conclusion HRQOL themes of CD have been identified and a preliminary HRQOL concept model has been established, which will provide a reference for the development of HRQOL evaluation tools in CD patients.
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Objective To study the chemical constituents from the stems and leaves of Monimopetalum chinense. Methods The constituents were isolated and purified by silica gel column chromatography and preparative HPLC. Their structures were elucidated based on spectroscopic methods including 1D and 2D NMR and HR-ESI-MS. Results Two new dihydro-β-agarofuran sesquiterpenes were isolated from the stems and leaves of M. chinense and identified as 1α,6β-dinicotinoyloxy-9α-acetoxy-dihydro- β-agarofuran (1) and 1α,6β-dinicotinoyloxy-9α-benzoyloxy-8α-hydroxydihydro-β-agarofuran (2). Conclusion Compounds 1 and 2 are new compounds, and named as monimins I and II, respectively.
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Induced pluripotent stem cells (iPSCs) have the potential of proliferation and differentiation into a variety of somatic cells, including hepatocyte-like cells (HLCs). HLCs from human iPSCs (hiPSC-HLCs) have similar features and functions as primary hepatocytes and are used as an efficient in vitro model of hepatocytes, which brings hope to studies on liver diseases and drug hepatotoxicity evaluation. This article reviews the research advances in hiPSC-HLCs and their application in the fields of disease model, drug hepatotoxicity evaluation, and cell transplantation and discusses the future perspectives of the application of hiPSC-HLCs.
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<p><b>OBJECTIVE</b>To explore the clinical characteristics, therapeutic measures and risk factors of pulmonary fungal infection in patients after renal transplantation.</p><p><b>METHODS</b>The clinical data of 176 patients receiving renal allograft transplantation with postoperative infections were retrospectively analyzed. Among the patients, 40 were diagnosed to have pulmonary fungal infection, and their clinical symptoms, signs, radiographic findings, pathogenic bacterial culture, histopathological examination, and treatments were analyzed.</p><p><b>RESULTS</b>The 40 recipients with postoperative pulmonary fungal infection included 25 male and 15 female patients with a mean age of 49 years. Twenty-eight of the patients developed pulmonary fungal infection within 6 months after transplantation. Positive pathogen cultivation was reported in 19 cases, and Candida albicans was detected in 11 cases, Candida krusei in 2 cases, Candida glabrata in 3 cases, Candida tropicalis in 1 case, aspergillosis in 1 case, and Candida mycoderma in 1 case. Twenty-four of out of the 40 cases were found to have co-infection. All the patients received antifungal drugs and adjuvant treatments, and 38 patients were cured and 2 died.</p><p><b>CONCLUSION</b>Pulmonary fungal infection often occurs within 6 months after renal transplantation. The most common fungal pathogen is Candida albicans, and the patients often had coinfections. Early diagnosis and timely intervention with antifungal drugs and comprehensive measures are critical in the management of pulmonary fungal infection following renal transplantation.</p>
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Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Antifúngicos , Usos Terapêuticos , Aspergillus , Candida , Transplante de Rim , Pneumopatias Fúngicas , Epidemiologia , Complicações Pós-Operatórias , Epidemiologia , Microbiologia , Estudos Retrospectivos , Fatores de RiscoRESUMO
Objective The radiotherapy resistance is one of important causes for nasopharyngeal carcinoma(NPC) treatment failure.Junctional adhesion molecule A(JAMA)is closely correlated with the tumor poor prognosis.Thus this experiment is to in vestigate the relationship between JAMA expression and the radiosensitivity of NPC.Methods To overexpress or interfere the JAMA expression in CNE2 and HONE1 cell lines.Then different doses of X-ray were adopted to conduct irradiation.The cell clone formation capacity and cellular apoptosis change were detected after 24 h.The role of JAMA in the NPC radiotherapy was understand.The related signal pathway protein in cell lines with different JAMA expression was detected by Western blot.Results The cell lines with low JAMA expression were more sensitive to radiotherapy:After low JAMA expression,the D0 value in the CNE2 cell line was decreased from 3.26 ±0.78 to 1.92 ± 0.23;the Dq value was decreased from 46.51 ± 4.27 to 32.12 ± 3.19.The radio therapy induced apoptosis was significantly increased in the cell lines with low JAMA expression,after low JAMA expressing,thcellular apoptosis was elevated from 6.9 % ± 0.9 % to 13.7 % ± 1.3 %;the HONE1 cellular apoptosis was elevated from 6.5 % + 1.1 % to 12.3 % ± 1.7%;JAMA overexpression cell lines were significantly decreased.The preliminary mechanism research results showed that JAMA played the effect via Akt signal pathway.Conclusion This research results verifiy that JAMA expression level is closely correlated with the radiosensitivity of NPC cell line:JAMA can increase the radiotherapy resistance of NPC cell lines,which provides a new feasible research direction for NPC enhancing radiosensitivity.
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Objective To identify the role of miR-124 in regulating the radiosensitivity and the epithelial-mesenchymal transition (EMT) of nasopharyngeal carcinoma (NPC). Methods Transient transfection of cells with miR-124 mimic or inhibitor was performed and wound-healing assay was used to investigate the role of miR-124 in the EMT of NPC. The apoptosis affected by miR-124 was also measured after irradiation , followed by investigating the cell proliferation by EdU assay. Finally , proteins of Akt and ERK associated with EMT and radiosensitivity, were measured by western blot. Results The migration index from NPC cell line indicated that miR-124 repressed the EMT. The results from caspase-3 activity assay showed that caspase-3 activity after irradiation significantly increased in miR-124 mimic group compared with the control group (P < 0.01). It was also confirmed that irradiation led to a higher percentage of apoptosis in miR-124 group compared with the control group in NPC cells. Cell proliferation after irradiation was significantly decreased in MiR-124 group as compared with control group . MiR-124 inhibited the protein expression of p-Akt . Conclusion MiR-124 may repress the EMT and decrease radio-resistance of NPC via p-Akt signaling pathway , which may provide a new insight into radio-resistance in NPC.
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To study the effect of the combined administration of different doses of Glycyrrhizae Radix et Rhizoma and Atractylodis Macrocephalae Rhizoma on the proliferation of DFMO-treated intestinal epithelial cells (IEC-6) and p53, p21 mRNA and protein expressions, in order to define the molecular basis for the effect of the combined administration of different doses of Glycyrrhizae Radix et Rhizoma and Atractylodis Macrocephalae Rhizoma on the cell proliferation. The effect of the drugs on the cell division rate and cell cycle of IEC-6 cells was detected by FCM. Quantitative Real-time PCR (qRT-PCR) was used to analyze the effect of the drugs on mRNA of p2l and p53 related to IEC-6 proliferation. Western blot was used to analyze the effect of the drugs on p2l and p53 protein expressions of IEC-6 cells. Atractylodis Macrocephalae Rhizoma could increase p53, p21 mRNA and proteins expression in DFMO-treated IEC-6 cells. The combined administration of different ratios of Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma could significantly down-regulate Atractylodis Macrocephalae Rhizoma's effect on p53, p21 mRNA and proteins expression in DFMO-treated IEC-6 cells and promote the proliferation of IEC-6 cells. The combined administration of Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma could down-regulate Atractylodis Macrocephalae Rhizoma's effect on DFMO-treated intestinal epithelial cells (IEC-6).
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Animais , Ratos , Atractylodes , Química , Linhagem Celular , Inibidor de Quinase Dependente de Ciclina p21 , Genética , Metabolismo , Medicamentos de Ervas Chinesas , Farmacologia , Células Epiteliais , Metabolismo , Expressão Gênica , Glycyrrhiza , Química , Intestinos , Metabolismo , Rizoma , Química , Proteína Supressora de Tumor p53 , Genética , MetabolismoRESUMO
Objective:To study the positive expression rate of M2 subtype of macrophage cell surface molecules and the inflammatory factors of PPS in IL-4-induced M2 macrophage.Methods:The experiment was divided into 5 groups:blank control group, Model group,PPS groups(50 μg/ml,100 μg/ml and 200 μg/ml).The expression of CD206 and CD23 was used as bio-maker to confirm IL-4 induced macrophages by treating RAW264.7 with 20ng/ml of IL-4.IL-4 induced RAW264.7 cells were treated with PPS of 50μg/ml,100μg/ml and 200μg/ml for 24 h.Then the expression of CD206,CD16/32 and CD40 were analyzed by flow cytometry, and the mRNA expression of IL-1β,TNF-α,IL-10 and iNOS were detect by qRT-PCR.Results: After treated with IL-4,the positive rate of CD206 of RAW264.7 were high.After treated with PPS ,the rate of CD16/32 and CD40 in IL-4 induced RAW264.7 cells were high ,the expression of CD206 decreased,and the mRNA level of IL-1βand TNF-αincreased.Conclusion:RAW264.7 cells can be polarlized to M2 subtype macrophage by using 20 ng/ml IL-4.PPS enhances the mRNA of IL-1β,TNF-αand the expression of CD40, CD16/32 in IL-4-induced RAW264.7 cells .These results indicate that PPS can induce the M2 subtype to become M1 macrophages, can improve immune function of macrophages.
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In traditional biomedical research, a series of mechanism and measures had been taken for identity protection of data subjects, such as data disclosure in aggregated methods, information restricted in public only after identified variables removal and etc. The purpose of such process was aimed to properly keep confidentiality of health information for the target subjects in research. As the protection of subject privacy was viewed as one of the most essential principle of medical ethics in human research, the effects to fulfill and accomplish such process can help to maintain the trust and support among participants and social public. Currently, such traditional modes of privacy safeguard are widely-applied in genetics and genomics study. However, the universal applicability also causes a number of controversies, and the effectiveness remains to be proven. Nowadays, the risk assessments of data subjects’ privacy call for taking the whole“data context” into consideration, not just self-restricted in isolation and confined to quality control of data disclosure. With the soaring increasing of data resources in research involved human subjects, the issues of releasing genetic data have caused more and more public attention, especially for the sensitive domains of privacy protection. Based on the core problem and principles, this article attempted to discuss the controversial bioethical issues such as data context, data-intruder concept, privacy of data subject, identity control of releasing data, potential risk of individual identification, privacy protection of data subject, and etc. We hope these considerations can provide references to the bioethical understanding of biobanks research and decision-making of ethic review.
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In order to study the excretion of genistein (GEN) capsule, an estrogen drugs, in human, 30 healthy volunteers were selected and orally administered 50, 100, and 300 mg genistein in an parallel study. Genistein were determined in urine by LC-MS/MS and glucuronidated genistein (GENG) were indirectly determined with enzymatic hydrolysis in urine by LC-MS/MS, and the pharmacokinetic parameters were analyzed by DAS software (ver 2.0). The result showed that the concentrations of genistein in human urine were less than 1% of the GENG, and the cumulative excretion of GEN in 48 h were 0.037, 0.134, and 0.142 mg, separately, and the urinary excretion percentage were only 0.07%, 0.13%, and 0.05%, separately. But the cumulative excretion of GENG in 48 h was 5.3, 13.8, and 15.4 mg, separately, and the urinary excretion percentage were 10.6%, 13.8%, and 5.1%, separately, and the max urinary excretive rate was 0.4, 1.0, and 1.4 mg x h(-1), separately (tmax were 6 h). Studies showed that part of drug excreted through kidney in a form of GENG in human, and the cumulative urinary excretion and the maximum excretion rate of GENG showed a proportional increase conditioned with the dose in the range of 50-100 mg, but showed non-linear increase feature in 300 mg.
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Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Administração Oral , Anticarcinógenos , Farmacocinética , Urina , Cromatografia Líquida , Genisteína , Farmacocinética , Urina , Glucuronídeos , Urina , Voluntários Saudáveis , Fitoestrógenos , Farmacocinética , Urina , Espectrometria de Massas em TandemRESUMO
In order to study the excretion of genistein (GEN) capsule, an estrogen drugs, in human, 30 healthy volunteers were selected and orally administered 50, 100, and 300 mg genistein in an parallel study. Genistein were determined in urine by LC-MS/MS and glucuronidated genistein (GENG) were indirectly determined with enzymatic hydrolysis in urine by LC-MS/MS, and the pharmacokinetic parameters were analyzed by DAS software (ver 2.0). The result showed that the concentrations of genistein in human urine were less than 1% of the GENG, and the cumulative excretion of GEN in 48 h were 0.037, 0.134, and 0.142 mg, separately, and the urinary excretion percentage were only 0.07%, 0.13%, and 0.05%, separately. But the cumulative excretion of GENG in 48 h was 5.3, 13.8, and 15.4 mg, separately, and the urinary excretion percentage were 10.6%, 13.8%, and 5.1%, separately, and the max urinary excretive rate was 0.4, 1.0, and 1.4 mg x h(-1), separately (tmax were 6 h). Studies showed that part of drug excreted through kidney in a form of GENG in human, and the cumulative urinary excretion and the maximum excretion rate of GENG showed a proportional increase conditioned with the dose in the range of 50-100 mg, but showed non-linear increase feature in 300 mg.
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<p><b>OBJECTIVE</b>To detect the effects of Polyporus polysaccharide (PPS), Bacillus Calmette-Guerin (BCG), and their combination on the nuclear factor kappa B (NF-κB) signaling pathway associated-gene expression and investigate the molecular mechanisms of the toxic-reducing effect of PPS in coordination with BCG against bladder cancer.</p><p><b>METHODS</b>After T739 cells were treated with PPS, BCG and their combination, the changes in mRNA and protein expression of inhibitor of kappa B kinase beta (IKKβ), NF-κB subunit p65 (NF-κB p65), intracellular adhesion molecule 1 (ICAM1) and chemokine (C-c motif) ligand 2 (CCL2) in bladder cancer cell line T739 were determined by relative quantitative real-time PCR, Western blot, and flow cytometry (FCM). NF-κB p65 DNA-binding activity in T739 cell was detected by biotinylated probe-ELISA, and NF-κB p65 nuclear expression in T739 cell was observed by immunohistochemistry.</p><p><b>RESULTS</b>Compared with the T739 control group, the mRNA expression of IKBKB (IKKβ), Rel A (NF-κB p65), ICAM1 and CCL2 in T739 cells treated with BCG were increased obviously (Ratio>2.0), as well as the expression of IKKβ, CCL2 and ICAM1 proteins. Meanwhile, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression in T739 cells treated with BCG were up-regulated significantly (P<0.05). Compared with the control, the increased expression in T739 cells were simultaneously down-regulated after PPS treatment, except for ICAM1 protein expression. With cells treated with a combination of BCG and PPS, the expression of genes associated with the NF-κB signaling pathway, such as IKBKB, ICAM1 and CCL2, were all down-regulated compared to the BCG group, as well as Rel A mRNA expression, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression.</p><p><b>CONCLUSIONS</b>PPS could inhibit the over-activation of the NF-κB signaling pathway induced by BCG in bladder cancer cells and accordingly attenuate the adverse reactions to BCG therapy.</p>
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Humanos , Linhagem Celular Tumoral , Núcleo Celular , Metabolismo , Regulação Neoplásica da Expressão Gênica , Mycobacterium bovis , NF-kappa B , Metabolismo , Proteínas de Neoplasias , Genética , Metabolismo , Polyporus , Química , Polissacarídeos , Farmacologia , Transdução de Sinais , Neoplasias da Bexiga Urinária , Genética , Microbiologia , PatologiaRESUMO
<p><b>OBJECTIVE</b>To study the effects of licorice on the proliferation of intestinal crypt stem cell line IEC-6 and the expression of p53.</p><p><b>METHODS</b>Induced by difluoro-methylornithine (DFMO), polyamine-depleted IEC-6 cells under growth inhibition were used as the pathological cell model in this study. Cells were divided into four groups, i. e., the control group, the DFMO-treated group, the high dose licorice group, and the low dose licorice group. The control group consisted of IEC-6 cells cultured in normal condition. The other three groups were all treated with 5 mmol/L DFMO. The high dose and low dose licorice groups were supplemented with 40 and 80 microg/mL licorice granule respectively. All the groups were cultured for 6 successive days. The cell number and viability were determined using flow cytometry. The level of p53 protein was detected by Western blot. The p53 mRNA levels and stability were detected using fluorescent quantitative Real-time PCR.</p><p><b>RESULTS</b>Compared with the control group, the cell growth of the DFMO group was obviously inhibited on the 4th day (P < 0.05). The cell number increased more obviously in the low dose licorice and the high dose licorice groups in a dose-dependent way on the 6th day when compared with the DFMO group (P < 0.05). When compared with the control group, significantly elevated expression levels of p53 protein and mRNA in cells of the DFMO group were detected after 6-day treatment (P < 0.05). When compared with the DFMO group, the expression levels of p53 protein and mRNA were significantly down-regulated in the low dose licorice and the high dose licorice groups (P < 0.05). The degradation of p53 mRNA was the fastest in the control group, while the degradation speed of cells in the DFMO group was the slowest.</p><p><b>CONCLUSION</b>One of mechanisms for protective and healing effects of licorice on the intestinal mucosa was possibly through down-regulating the stability of p53 mRNA, lowering the expression of p53, thus promoting the proliferation of the intestinal crypt stem cells.</p>