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The canonical transient receptor potential channel(TRPC)proteins form Ca2+-permeable cation channels that are involved in various heart diseases.However,the roles of specific TRPC proteins in myocardial ischemia/reperfusion(I/R)injury remain poorly understood.We observed that TRPC1 and TRPC6 were highly expressed in the area at risk(AAR)in a coronary artery ligation induced I/R model.Trpc1-/-mice exhibited improved cardiac function,lower serum Troponin T and serum creatine kinase level,smaller infarct volume,less fibrotic scars,and fewer apoptotic cells after myocardial-I/R than wild-type or Trpc6-/-mice.Cardiomyocyte-specific knockdown of Trpc1 using adeno-associated virus 9 mitigated myocardial I/R injury.Furthermore,Trpc1 deficiency protected adult mouse ventricular myocytes(AMVMs)and HL-1 cells from death during hypoxia/reoxygenation(H/R)injury.RNA-sequencing-based transcriptome analysis revealed differential expression of genes related to reactive oxygen species(ROS)generation in Trpc1-/-cardiomyocytes.Among these genes,oxoglutarate dehydrogenase-like(Ogdhl)was markedly downregulated.Moreover,Trpc1 deficiency impaired the calcineurin(CaN)/nuclear factor-kappa B(NF-κB)signaling pathway in AMVMs.Suppression of this pathway inhibited Ogdhl upregulation and ROS generation in HL-1 cells under H/R conditions.Chromatin immunoprecipitation assays confirmed NF-κB binding to the Ogdhl promoter.The cardioprotective effect of Trpc1 deficiency was canceled out by overexpression of NF-κB and Ogdhl in cardiomyocytes.In conclusion,our findings reveal that TRPC1 is upregulated in the AAR following myocardial I/R,leading to increased Ca2+influx into associated cardiomyocytes.Subsequently,this upregulates Ogdhl expression through the CaN/NF-κB signaling pathway,ultimately exacerbating ROS production and aggravating myocardial I/R injury.
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OBJECTIVE@#To investigate the clinicopathologic features and prognostic factors in oral basaloid squamous cell carcinoma.@*METHODS@#Retrospective analysis of oral basaloid squamous cell carcinomas patients who underwent tumor resection during the period from January 2002 to December 2020 in the authors' hospital, especially the clinicopathologic characteristics of 28 cases with confirmed diagnosis and follow-up data. Immunohistochemistry was performed to define the helpful markers for differentiation diagnosis. The factors influencing the prognosis were evaluated based on Kaplan-Meier method.@*RESULTS@#The tongue and mouth floor (11 cases, 39.3%) were the most frequently involved sites, followed by gingiva (6 cases, 21.4%), buccal (5 cases, 17.9%), palate (4 cases, 14.3%), and oropharynx (2 cases, 7.1%). The majority of basaloid squamous cell carcinomas were in advanced stage, with 12 cases in stage Ⅱ and 16 cases in stages Ⅲ-Ⅳ. Twelve of 28 patients were identified to have cervical lymph node metastasis, which was confirmed by histopathological examination. The incidence rate of lymph node metastasis was 42.9% (12/28). Nine tumors recurred, with one metastasized to the lung. At the meantime, the 28 conventional squamous cell carcinomas were matched with the same stage, among which 13 cases were identified with cervical lymph node metastasis. The incidence rate of lymph node metastasis was 46.4% (13/28). Five cases recurred, with two cases that metastasized to the lung and one to the brain. The 5-year overall survival rates of the basaloid squamous cell carcinoma and conventional squamous cell carcinoma patients were 54.6% and 53.8%, respectively. Histopathologically, basaloid cells consisted of tumor islands without evident keratinization but frequently with comedo-like necrosis within the tumor islands. CK5/6 and P63 exhibited strongly positive in all the 28 cases, whereas neuroendocrine markers, CgA and Syn, were negative. Eight cases positively expressed P16; one case showed focal SOX10 positive but CK7 negative.@*CONCLUSION@#The majority of oral basaloid squamous cell carcinomas present in advanced stage with a high tendency to lymph node metastasis, but the overall survival rates are not significantly different from conventional squamous cell carcinomas matched with the same stage. The human papilloma virus (HPV), as HPV-positivity rate is high, correlates to good prognosis. In addition, CK7 & SOX10 immunohistochemistry could contribute to differential diagnosis for basaloid squamous cell carcinoma with solid adenoid cystic carcinoma.
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Humanos , Carcinoma Adenoide Cístico , Carcinoma de Células Escamosas , Imuno-Histoquímica , Recidiva Local de Neoplasia/epidemiologia , Estudos RetrospectivosRESUMO
Alzheimer's disease(AD) patients in China have been surging, and the resultant medical burden and care demand have a huge impact on the development of individuals, families, and the society. The active component compound of Epimedii Folium, Astragali Radix, and Puerariae Lobatae Radix(YHG) can regulate the expression of iron metabolism-related proteins to inhibit brain iron overload and relieve hypofunction of central nervous system in AD patients. Hepcidin is an important target regulating iron metabolism. This study investigated the effect of YHG on the expression of a disintegrin and metalloprotease-17(ADAM17), a key enzyme in the hydrolysis of β amyloid precursor protein(APP) in HT22 cells, by mediating hepcidin. To be specific, HT22 cells were cultured in vitro, followed by liposome-mediated siRNA transfection to silence the expression of hepcidin. Real-time PCR and Western blot were performed to examine the silencing result and the effect of YHG on hepcidin in AD cell model. HT22 cells were randomized into 7 groups: control group, Aβ25-35 induction(Aβ) group, hepcidin-siRNA(siRNA) group, Aβ25-35 + hepcidin-siRNA(Aβ + siRNA) group, Aβ25-35+YHG(Aβ+YHG) group, hepcidin-siRNA+YHG(siRNA+YHG) group, Aβ25-35+hepcidin-siRNA+YHG(Aβ+siRNA+YHG) group. The expression of ADAM17 mRNA in cells was detected by real-time PCR, and the expression of ADAM17 protein by immunofluorescence and Western blot. Immunofluorescence showed that the ADAM17 protein expression was lower in the Aβ group, siRNA group, and Aβ+siRNA group than in the control group(P<0.05) and the expression was lower in the Aβ+siRNA group(P<0.05) and higher in the Aβ+YHG group(P<0.05) than in the Aβ group. Moreover, the ADAM17 protein expression was lower in the Aβ+siRNA group(P<0.05) and higher in the siRNA+YHG group(P< 0.05) than in the siRNA group. The expression was higher in the Aβ+siRNA+YHG group than in the Aβ+siRNA group(P<0.05). The results of Western blot and real-time PCR were consistent with those of immunofluorescence. The experiment showed that YHG induced hepcidin to up-regulate the expression of ADAM17 in AD cell model and promote the activation of non-starch metabolic pathways, which might be the internal mechanism of YHG in preventing and treating AD.
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Humanos , Proteína ADAM17 , Doença de Alzheimer/genética , Peptídeos beta-Amiloides , Medicamentos de Ervas Chinesas/farmacologia , Hepcidinas/genética , PuerariaRESUMO
OBJECTIVE@#To investigate the incidence and clinicopathologic features of cervical lymph node metastasis in salivary gland adenoid cystic carcinoma (AdCC).@*METHODS@#Retrospective analysis was made in 798 AdCC patients who underwent tumor resection during January, 2001 to January, 2019 in Peking University School and Hospital of Stomatology, especially the clincopathologic characteristics of 82 cases with lymph node metastasis.@*RESULTS@#In the study, 82 of the 798 patients were identified with cervical lymph node metastasis, which was confirmed by histopathological examination. The palate, the region of sublingual gland and mouth floor, and the parotid were the frequently involved primary sites for AdCCs. The general incidence rate of lymph node metastasis was approximately 10%. The submandibular gland, the region of sublingual gland and mouth floor, and the mobile tongue were the most frequent sites of lymph node metastasis with the incidence rates of 20.8%, 16.1%, and 15.1%, respectively; while lymph node metastasis was uncommon in the tumors which were the origin from the palate and parotid, with incidence rates of 6.1% and 3.4%, respectively. Most AdCC cases (70.7%) showed the classic "tunnel-style" metastatic pattern of occurrence, and the level I and II regions were the most frequently involved areas. The 5-year and 10-year overall survival rates of the patients with lymph node metastasis were 77.4% and 20.6% respectively, while the 5-year and 10-year overall survival rates of the patients with no lymph node metastasis were 83.5% and 57.6%, respectively. The univariate analysis demonstrated that statistically significant differences in the overall survival for the presence of lymph node metastasis (P<0.001). In the meantime, the 5-year disease-free survival rate of the patients with lymph node metastasis also showed statistically significant differences to that of the AdCC patients with no lymph node metastasis. In addition, the primary site and histological grade were significantly associated with lymph node metastasis, and the high-grade solid growth pattern was identified as a strong predictor for the occurrence of lymph node metastasis.@*CONCLUSION@#Cervical lymph node metastasis has a high tendency of occurrence in submandibular gland and tongue-mouth floor complex, and the high-grade solid growth pattern could be taken as a strong predictor for the occurrence of lymph node metastasis, which correlates to poor prognosis of AdCC patients. A selective neck dissection should be considered as a management in such patients.
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Humanos , Carcinoma Adenoide Cístico , Linfonodos , Metástase Linfática , Estudos Retrospectivos , Neoplasias das Glândulas Salivares , Glândulas SalivaresRESUMO
There are many problems in the use of Bupleurum medicinal materials such as confusion in variety, unclearness in origin and quality problems, e.g., adulteration of authentic Bupleurum and mixing of non-medicinal parts. Therefore, the quality of Chinese patent medicines containing Bupleurum chinense is variable, and it is necessary to establish targeted quality control methods. Based on the quality control methods of Bupleurum and its Chinese patent medicines, combined with the example of Xiaochaihu granules, this paper discussed the quality standard improvement and supplementary test methods of Bupleurum and its Chinese patent medicines, aiming to provide reference for the quality control, quality supervision and standardized production of Chinese patent medicines containing Bupleurum and promote the improvement of its intrinsic quality.
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The chemical property of cinnamaldehyde is unstable in vivo, although early experiments have shown its obvious therapeutic effects on viral myocarditis (VMC). To overcome this problem, we used cinnamaldehyde as a leading compound to synthesize derivatives. Five derivatives of cinnamaldehyde were synthesized: 4-methylcinnamaldehyde (1), 4-chlorocinnamaldehyde (2), 4-methoxycinnamaldehyde (3), α-bromo-4-methylcinnamaldehyde (4), and α-bromo-4-chlorocinnamaldehyde (5). Neonatal rat cardiomyocytes and HeLa cells infected by coxsackievirus B3 (CVB3) were used to evaluate their antiviral and cytotoxic effects. In vivo BALB/c mice were infected with CVB3 for establishing VMC models. Among the derivatives, compound 4 and 5 inhibited the CVB3 in HeLa cells with the half-maximal inhibitory concentrations values of 11.38 ± 2.22 μM and 2.12 ± 0.37 μM, respectively. The 50% toxic concentrations of compound 4 and 5-treated cells were 39-fold and 87-fold higher than in the cinnamaldehyde group. Compound 4 and 5 effectively reduced the viral titers and cardiac pathological changes in a dose-dependent manner. In addition, compound 4 and 5 significantly inhibited the secretion, mRNA and protein expressions of inflammatory cytokines TNF-α, IL-1β and IL-6 in CVB3-infected cardiomyocytes, indicating that brominated cinnamaldehyde not only improved the anti-vital activities for VMC, but also had potent anti-inflammatory effects in cardiomyocytes induced by CVB3.
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Animais , Humanos , Camundongos , Ratos , Citocinas , Células HeLa , Interleucina-6 , Miocardite , Miócitos Cardíacos , RNA Mensageiro , Usos TerapêuticosRESUMO
Objective: A rapid and specific ultra performance liquid chromatography (UPLC) method was established for simultaneous analysis on six compounds and fingerprint analysis on Polygalae Radix to evaluate the herb quality from different habitats in China. Methods: The UPLC method was carried out by gradient elution with acetonitrile-formic acid water (0.1%). The flow rate was 0.4 mL/min. The detection wavelength was at 320 nm. The fingerprint chromatograms and the contents of six compounds including sibiricose A5, sibiricose A6, sibiricaxanthone B, glomeratose A, polygalaxanthone III, and 3,6'-disinapoyl sucrose in 24 batches of Polygalae Radix were analyzed. The common peaks were identified by ultra performance liquid chromatography tandem with time-of-flight mass spectrometry with MSE data-acquistion mode (UPLC-Q-TOF-MSE). Results: There was a difference in contents of six compounds, especially for the content of 3,6'-disinapoyl sucrose and sibiricose A6. Thirty-seven peaks were selected as the common peaks, of which 33 peaks were identified, and the similarities of 24 batches were between 0.756 and 0.997. Based on the results of quantification and fingerprint analysis, a certain difference between samples from different habitats was further proven. Conclusion: The validated UPLC quantitative analysis and fingerprint methods are successfully used in the quality control of Polygalae Radix.
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Objective: To establish an HPLC fingerprint of Xiangsha Liujun Pills (XLP), and to apply this method to determining 49 batches of samples from 16 different manufacturers for quality evaluation, supporting the foundation of quality control system of XLP. Methods: The HPLC separation was performed on a Sepax Bio-C18 reversed-phase column in gradient mode of acetonitrile-0.1% acetic acid with UV detection at 260 nm. The column temperature was kept at 30℃, and the flow rate of mobile phase was 1.0 mL/min. The fingerprint similarities and cluster analysis based on the chromatographic peaks in detected samples were obtained. Results: The HPLC fingerprint of XLP was developed firstly. Seventeen common fingerprint peaks were indicated and attributed to raw materials, and 16 of them were identified by comparison with reference standards and UPLC-Q-TOF-MS. The analyzed samples were geographically classified into four classes with the similarities between 0.617-0.988. Conclusion: Significant differences of quality in XLP on the market are observed. The method is simple, reliable, and suitable for the quality control of XLP, supplying helpful information for the comprehensive evaluation of XLP.
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<p><b>OBJECTIVE</b>To study the diagnostic value and influencing factors for amplitude-integrated EEG (aEEG) in brain injury in preterm infants.</p><p><b>METHODS</b>One hundred and sixteen preterm infants with a gestational age (GA) between 27 weeks and 36(+6) weeks were enrolled as subjects. The aEEG scores of all preterm infants were obtained within 6 hours after birth. According to the diagnostic results, the 116 preterm infants were divided into two groups: brain injury (n=63) and non-brain injury (n=53). The risk factors for brain injury were evaluated using logistic regression analysis. According to the aEEG results, the 116 preterm infants were divided into two groups: normal aEEG (n=58) and abnormal aEEG (n=58). The influencing factors for aEEG results in preterm infants were determined using univariate analysis.</p><p><b>RESULTS</b>The brain injury group had a significantly higher rate of abnormal aEEG than the non-brain injury group (83% vs 11%; P<0.05). The infants in the brain injury group from two different GA subgroups (27-33(+6) weeks and 34-36(+6) weeks) had significantly lower aEEG scores than the non-brain injury group from corresponding GA subgroups (P<0.01). Logistic regression analysis showed that low GA (<32 weeks), low birth weight (<1 500 g), abnormal placenta, fetal membranes, and umbilical cord, and hypertension during pregnancy were high-risk factors for brain injury (P<0.05). There were significant differences in GA, birth weight, abnormal placenta, fetal membranes, and umbilical cord, and hypertension during pregnancy between the normal and abnormal aEEG groups (P<0.05).</p><p><b>CONCLUSIONS</b>The risk factors for brain injury are consistent with the influencing factors for aEEG results in preterm infants, suggesting that aEEG contributes to the early diagnosis of brain injury.</p>
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Feminino , Humanos , Recém-Nascido , Gravidez , Peso ao Nascer , Lesões Encefálicas , Diagnóstico , Eletroencefalografia , Recém-Nascido Prematuro , Modelos Logísticos , Fatores de RiscoRESUMO
Twelve compounds were isolated from the rhizome of Paris mairei Lévl by silica gel, Sephadex LH-20 and ODS col-umn chromatographies. The structure elucidation was accomplished by ESI-MS and NMR methods. These compounds were identified as lupeol(1), lup-20(29) -ene-3β-yl octacosanoate(2), palmitic acid(3), glyceryl α-mono-palmitate(4), α-spinasterol(5), diosgenin (6), (25R) diosgenin-3-O-α-L-rhamnopyranosyl (1--> 4) -α-L-rhamnopyranosyl (1 --> 4) - [α-L-rhamnopyranosyl(1 --> 2)] -β-D-glucopyranoside(7), pennogenin(8), pennogenin-3-O-β-D-glucopyranosyl(1 -->3) - [α-L-rhamnopyranosyl(1 --> 2)] -β-D-glucopyranoside(9), flazin(10), calonysterone(11), and isorhamnetin-3-O-β-gentiobioside(12). Compounds 1-5,10-11 were isolated from the genus Paris for the first time, and all compounds were isolated from this plant for the first time.
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Medicamentos de Ervas Chinesas , Química , Liliaceae , Química , Estrutura Molecular , Rizoma , Química , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
<p><b>OBJECTIVE</b>To study the pharmacokinetics of a novel recombinant human granulocyte colony-stimulating factor (rhG-CSFa) in rats and to determine the proteolytic rates of rhG-CSFa in the whole blood and serum of rats in vitro.</p><p><b>METHODS</b>The pharmacokinetics of rhG-CSFa and conventional (wild type, WT) granulocyte colony-stimulating factor (G-CSF) were investigated in Sprague-Dawley rats which received either intravenous or subcutaneous injection of rhG-CSFa or WT G-CSF at three different doses (20, 50, or 100 microg/kg). The blood samples of rats were collected at multiple time points (from 0.08 to 12 h) and the concentrations of rhG-CSFa and WT G-CSF in serum were determined with a sandwich enzyme-linked immunosorbent assay (ELISA). For the study of proteolytic rates in vitro, the concentrations of rhG-CSFa or WT G-CSF were determined at 3-minute intervals after addition of the respective drug to rat's whole blood or serum.</p><p><b>RESULTS</b>Pharmacokinetic analysis of serum rhG-CSFa or WT G-CSF levels indicated that, at each dose tested, for either route of drug administration, the area under concentration-time curve values and the maximum serum concentration of rhG-CSFa were higher than those of WT G-CSF, and the serum half life of rhG-CSFa was longer than that of WT G-CSF. Subsequent in vitro whole blood and serum stability study showed that the rates of drug degradation in WT G-CSF were 1.8 folds and 1.5 folds higher than those in rhG-CSFa, respectively.</p><p><b>CONCLUSION</b>rhG-CSFa has better serum and whole blood stability in vitro and higher bioavailability in vivo as compared to WT G-CSF.</p>
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Animais , Feminino , Humanos , Masculino , Camundongos , Ratos , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Métodos , Padrões de Referência , Fator Estimulador de Colônias de Granulócitos , Sangue , Genética , Farmacocinética , Camundongos Endogâmicos BALB C , Ratos Sprague-Dawley , Proteínas Recombinantes , Sensibilidade e EspecificidadeRESUMO
<p><b>OBJECTIVE</b>To investigate the chemical constituents from the root of Psammosilene tunicoides.</p><p><b>METHOD</b>Column chromatographic methods were used to isolate the chemical constituents of this plant. ESI-MS, EI-MS and NMR methods were employed for their structural elucidation.</p><p><b>RESULT</b>Seven compounds were isolated and identified as goyaprosaponin (1), Soya-cerebroside (2), tectoridin (3), alpha-spinasterol (4), tetracosanoic acid (5), beta-sitosterol (6), daucosterol (7) respectively.</p><p><b>CONCLUSION</b>Compounds 2-7 were obtained from genus Psammosilene for the first time.</p>