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Traditional Chinese medicine(TCM) has a long history and abundant experience in external therapy, which marks human wisdom. In the early history of human, people found that fumigation, coating, and sticking of some tree branches and herb stems can help alleviate scabies and remove parasites in productive labor, which indicates the emergence of external therapy. Pathogen usually enters the body through the surface, so external therapy can be used to treat the disease. External therapy is among the major characteristic of surgery of TCM. As one of the external therapies in TCM, external application to acupoints smooths the zang-fu organs through meridians and collaterals, thereby harmonizing yin and yang. This therapy emerged in the early society, formed the Spring and Autumn Period and the Warring States Period, improved in the Song and Ming dynasties, and matured in the Qing dynasty. With the efforts of experts in history, it has had a mature theory. According to modern research, it can avoid the first-pass effect of liver and the gastrointestinal irritation and improve the bioavailability of Chinese medicine. Based on the effect of Chinese medicine and the theory of meridian and collateral, it can stimulate the acupoints, exert regulatory effect on acupoints, and give full play to the efficacy of TCM and the interaction of the two. Thereby, it can regulate qi and blood and balance yin and yang, thus being widely used in the treatment of diseases. In this paper, the use of external application to acupoints, the effect on skin immunity, the regulation of neuro-inflammatory mechanism, the relationship between acupoint application and human circulation network, and the development of its dosage form were summarized through literature review. On this basis, this study is expected to lay a foundation for further research.
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Puntos de Acupuntura , Meridianos , Humanos , Disponibilidad Biológica , Fumigación , Medicina Tradicional ChinaRESUMEN
In this experiment, Panax notoginseng saponins chitosan nanoparticles(PNS-NPs) were prepared by self-assembly and their appearance, particle size, encapsulation efficiency, drug loading, polydispersity index(PDI), Zeta potential, and microstructure were characterized. The prepared PNS-NPs were intact in structure, with an average particle size of(209±0.258) nm, encapsulation efficiency of 42.34%±0.28%, a drug loading of 37.63%±0.85%, and a Zeta potential of(39.8±3.122) mV. The intestinal absorption of PNS-NPs in rats was further studied. The established HPLC method of PNS was employed to investigate the effects of pH, perfusion rate, and different drugs(PNS raw materials, Xuesaitong Capsules, and PNS-NPs). The absorption rate constant(K_a) and apparent permeability coefficient(P_(app)) in the duodenum, jejunum, ileum, and colon were calculated and analyzed. As illustrated by the results, the intestinal absorption of PNS-NPs was increased in the perfusion solution at pH 6.8(P<0.05), and perfusion rate had no significant effect on the K_a and P_(app) of PNS-NPs. The intestinal absorption of PNS-NPs was significantly different from that of PNS raw materials and Xuesaitong Capsules(P<0.05), and the intestinal absorption of PNS-NPs was significantly improved.
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Quitosano , Nanopartículas , Panax notoginseng , Saponinas , Animales , Quitosano/farmacología , Absorción Intestinal , Panax notoginseng/química , Ratas , Saponinas/farmacologíaRESUMEN
OBJECTIVE: To understand the characteristics of DNA methyltransferase 3a (DNMT3a) in thymoma associated Myasthenia Gravis reveal its transcriptional regulator network as while as analyze the effect of DNMT3a on Rel/ nuclear factor-kappaB family (RelA/RelB) and its downstream autoimmune regulatory factor (Aire). METHODS: Tissues of 30 patients with thymoma, with or without myasthenia gravis (MG), were collected and the DNMT3a protein expression were evaluated through immunohistochemistry. We performed mRNA expression profiling microarray detection and analysis, and integrated the analysis by constructing protein-protein interaction networks and the integration with other database. We identified molecular difference between low and high DNMT3a in the thymoma by heatmap. We also performed PCR validation in thymoma tissues. The DNMT3a-shRNA plasmid was transfected into TEC cells, and these cells were treated with 5-aza-2-deoxycytidine, a blocker of DNMT3a. After the down-regulation of DNMT3a in TEC cells, the transcript and protein levels of RelA, RelB, Aire, and CHRNA3 were evaluated by western blotting. In addition, changes in gene expression profiles were screened through microarray technology. We performed differential gene analysis in the thymoma cohort by heatmap with R (v.4.3.0) software. RESULTS: In 30 matched tissue specimens, the expression of DNMT3a protein in thymoma with MG was lower than that in thymoma. Through mRNA expression profiling analysis, we constructed a co-expression network of DNMT3a and found direct interaction between IKZF1 and DNMT3a, and this co-expression relationship was overlappted with Cistrome DB database. We found up-regulation of 149 mRNAs and repression of 177 mRNAs in thymoma with MG compared with thymoma. Gene ontology and pathway analysis show the involvement of a multitude of genes in the mis-regulation of MG-related pathways. RNA interference significantly reduced the level of mRNA of DNMT3a, which proved that plasmid DNMT3a was effective. In comparison to the control group, the levels of DNMT3a, Aire, and CHRNA3 mRNA and protein in TEC cells transfected with DNMT3a-shRNA interference plasmid were significantly decreased, while the expression level of RelA and RelA/RelB was significantly increased. CONCLUSIONS: Our study reveals the DNMT3a-NF-κB pathway has a major effect on MG, and can be used as a marker for diagnosis as well as a target for MG treatment.
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ADN Metiltransferasa 3A/biosíntesis , Células Epiteliales/metabolismo , Regulación Neoplásica de la Expresión Génica , Miastenia Gravis/metabolismo , FN-kappa B/biosíntesis , Proteínas de Neoplasias/biosíntesis , Interferencia de ARN , Timoma/metabolismo , Timo/metabolismo , Neoplasias del Timo/metabolismo , Adolescente , Adulto , ADN Metiltransferasa 3A/antagonistas & inhibidores , ADN Metiltransferasa 3A/genética , Decitabina/farmacología , Ontología de Genes , Humanos , Masculino , Persona de Mediana Edad , Miastenia Gravis/etiología , Miastenia Gravis/genética , FN-kappa B/genética , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/genética , Mapas de Interacción de Proteínas , ARN Neoplásico/genética , ARN Interferente Pequeño/genética , Receptores Nicotínicos/biosíntesis , Receptores Nicotínicos/genética , Timoma/complicaciones , Timoma/genética , Neoplasias del Timo/complicaciones , Neoplasias del Timo/genética , Análisis de Matrices Tisulares , Factores de Transcripción/biosíntesis , Factores de Transcripción/genética , Transcriptoma , Proteína AIRERESUMEN
Non-small cell lung cancer (NSCLC) is the leading cause of cancer-associated mortality. Therapies targeting programmed cell death 1 ligand 1 (PD1L1) have promising effects on NSCLC. However, resistance to targeted therapy has become the main problem and the underling molecular mechanism remains unclear. In the present study, the expression of PD1L1 in NSCLC was determined and the association with clinicopathological characteristics was analyzed. A combination therapy was also constructed, including pembrolizumab (Pem) and iodine-125 (125I), which represented an efficient strategy for the treatment of NSCLC. The expression of PD1L1 was upregulated in NSCLC tissues and positively correlated with the Ki-67 index, pathological subtypes and risk stages. A higher level of PD1L1 expression was associated with poorer survival in patients with NSCLC, which could be used as a prognostic indicator. When NSCLC cells were cultured in the presence of Pem and 125I seeds, the combination treatment significantly abrogated the tumor proliferation and aggressiveness through the inhibition of matrix metalloproteinase-2 and -9 secretion. Flow cytometry analysis revealed pembrolizumab combined with 125I contributed to a higher rate of apoptosis and cell cycle arrest, indicating that the combination treatment improved the resistance to immunotherapy. Furthermore, the associated molecular mechanism was the dysregulation of ADAM metallopeptidase domain 17. The findings from the present study revealed that PD1L1 could be used as a predictive biomarker, and the application of combination treatment of pembrolizumab and 125I showed promising effects on NSCLC.
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In this paper, as an active ingredient, puerarin chitosan nanoparticles (Pur-CS/TPP-NPs) are prepared by an ionic gelation method. The chitosan (CS) concentration, pH of the CS solution, sodium tripolyphosphate (TPP) concentration, stirring speed, stirring time, ultrasonic power, and dosage are used as single factors for investigation, and the encapsulation efficiency, drug loading capacity, particle size, and polydispersity index (PDI) are used as indicators for investigation. The optimal prescription is determined using the Box-Behnken effect surface design method. The characterization of the best formulation, which is determined via an in vitro release assay and liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis methods, is used here for pharmacokinetic studies. An in situ single-pass intestinal perfusion model is used to investigate drug absorption in the intestine. After characterization, the morphologies of the nanoparticles are intact. It can be seen from the in vitro release experiments that the equation fitted by the nanoparticles is the Higuchi model, the nanoparticle release process is very stable and without sudden release, indicating that the nanoparticles are well-released in vitro. The pharmacokinetic results and the in situ single-pass intestinal perfusion model study show that the bioavailability and absorption of Pur-CS/TPP-NPs were significantly higher than Pur. Thus, all the results show that the prepared nanoparticles can significantly improve the bioavailability of Pur, and we hope to lay the foundation for the development of new products of Pur.
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BACKGROUND: This study was conducted to investigate the gene expression profiles associated with thymoma to better understand the molecular mechanism underlying the pathogenesis of thymoma. METHODS: Eight patients with thymomas (type A, AB, B1, and B2) and four controls with thymic cysts were analyzed using microarray profiling to identify changes in gene expression. RESULTS: Across all of our samples, 2319 messenger RNAs were upregulated and 2776 were downregulated in thymomas relative to thymic cysts. Gene ontology and pathway analyses revealed that a large number of genes participate in cellular functions, among which MHC class II protein complex assembly, assembly with peptide antigen, calcium activated phosphatidylcholine scrambling, and release of cytoplasmic sequestered NF-κB were dysregulated, whereas intestinal immune network for immunoglobulin A production, cytokine-cytokine receptor interaction, the calcium signaling pathway, and pathways related to autoimmune diseases were downregulated. CONCLUSIONS: Our results revealed gene expression differences between thymomas and thymic cysts, and identified key candidate genes/pathways that might be used as diagnostic markers and potential therapeutic targets to treat cancer metastasis.
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Regulación Neoplásica de la Expresión Génica , Timoma/genética , Transcriptoma , Biomarcadores de Tumor , Estudios de Casos y Controles , Biología Computacional/métodos , Femenino , Perfilación de la Expresión Génica , Ontología de Genes , Humanos , Masculino , Quiste Mediastínico/genética , Timoma/patologíaRESUMEN
BACKGROUND: To investigate the gene expression profile of a set of candidate genes for a better understanding of the molecular mechanism underlying the pathogenesis of thymoma with or without myasthenia gravis. METHODS: Thymoma patients and thymoma patients with myasthenia gravis were analyzed using microarray profiling to identify significant changes in gene expression of autoimmune regulator pathway genes including AIRE, IL-7R, CHRNA3, SYMD1, THRA, and CAV3. RESULTS: Across all of our samples, we found that 1484 mRNAs were upregulated and 770 were downregulated in thymoma patients compared with thymoma with myasthenia gravis patients. Gene ontology and pathway analysis revealed that a large number of genes participated in cellular functions for humoral immune response, sequence-specific DNA binding RNA polymerase II transcription factor activity, positive regulation of gene expression, regulation of neuron projection development, extracellular ligand-gated ion channel activity, positive regulation of striated muscle cell differentiation, and regulation of nuclear factor-kappaB import into the nucleus. CONCLUSION: Our results revealed genetic differences between thymomas and myasthenia gravis, and identified the key candidate genes/pathways for molecular mechanism.
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Miastenia Gravis/genética , Proteínas de Neoplasias/genética , Timoma/genética , Transcriptoma/genética , Adulto , Enfermedades Autoinmunes/complicaciones , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Caveolina 3/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Masculino , Análisis por Micromatrices , Persona de Mediana Edad , Miastenia Gravis/complicaciones , Miastenia Gravis/patología , FN-kappa B/genética , Receptores de Interleucina-7/genética , Receptores Nicotínicos/genética , Timoma/complicaciones , Timoma/patología , Receptores alfa de Hormona Tiroidea/genética , Factores de Transcripción/genética , Proteína AIRERESUMEN
BACKGROUND: Recombined humanized endostatin (Rh-endostatin) exhibits a potent anti-cancer effect involving multiple molecular targets and signaling pathways. HMGB1 is a highly conserved DNA-binding protein involved in cancer development. The therapeutic effect of Rh-endostatin on HMGB1 has not been reported, thus we investigate the effect in non-small cell lung cancer (NSCLC) cells. METHODS: Quantitative real-time PCR and Western blot were used to analyze the messenger RNA and protein expression of HMGB1 in A549 cancer cells, while enzyme-linked immunosorbent assay was used to detect the release of HMGB1. Western blot was performed to evaluate HMGB1 expression in SK-MES-1 and H661 NSCLC cells. RESULTS: Rh-endostatin inhibited the proliferation of A549 cancer cells and distinctly downregulated the expression and release of HMGB1 in dose and time dependent manners. Rh-endostatin-induced HMGB1 downregulation was confirmed in different types of NSCLC cells. CONCLUSION: These results demonstrate the general phenomenon that Rh-endostatin can induce HMGB1 suppression in a variety of NSCLC cells. Rh-endostatin may suppress HMGB1 expression and release in A549 cancer cells, thus inhibiting cell proliferation.
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Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Endostatinas/farmacología , Proteína HMGB1/genética , Proteínas Recombinantes/farmacología , Células A549 , Apoptosis/efectos de los fármacos , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Proliferación Celular/efectos de los fármacos , Endostatinas/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Proteína HMGB1/antagonistas & inhibidores , Humanos , Proteínas Recombinantes/genética , Transducción de Señal/efectos de los fármacosRESUMEN
BACKGROUND: Computed tomography (CT)-guided percutaneous implantation of 125 Iodine radioactive seeds requires the precise arrangement of seeds by tumor shape. We tested whether selecting target areas, including subclinical areas around tumors, can influence locoregional recurrence in patients with non-small cell lung cancer (NSCLC). METHOD: We divided 82 patients with NSCLC into two groups. Target areas in group 1 (n = 40) were defined along tumor margins based on lung-window CT. Target areas in group 2 (n = 42) were extended by 0.5 cm in all dimensions outside tumor margins. Preoperative plans for both groups were based on a treatment plan system, which guided 125 I seed implantation. Six months later, patients underwent chest CT to evaluate treatment efficacy (per Response Evaluation Criteria in Solid Tumors version 1). We compared locoregional recurrences between the groups after a year of follow-up. We then used the treatment plan system to extend target areas for group 1 patients by 0.5 cm (defined as group 3 data) and compared these hypothetical group 3 planned seeds with the actual seed numbers used in group 1 patients. RESULTS: All patients successfully underwent implantation; none died during the follow-up period. Recurrence was significantly lower in group 2 than in group 1 ( P < 0.05). Group 1 patients and group 3 data significantly differed in seed numbers ( P < 0.01). CONCLUSION: Our results imply that extending the implantation area for 125 I seeds can decrease recurrence risk by eradicating cancerous lymph-duct blockades within the extended areas.
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Braquiterapia/efectos adversos , Carcinoma de Pulmón de Células no Pequeñas/radioterapia , Radioisótopos de Yodo/administración & dosificación , Recurrencia Local de Neoplasia/radioterapia , Anciano , Carcinoma de Pulmón de Células no Pequeñas/patología , Femenino , Humanos , Radioisótopos de Yodo/efectos adversos , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia/patología , Radioinmunoterapia , Tomografía Computarizada por Rayos XRESUMEN
HPLC-ELSD was applied to explore the absorption mechanism of pulchinenosides (B3, BD, B7, B10, B11) in rats. The experimental results showed that the absorption rate constant, Ka value (B3, BD) and Permeability coefficient, Peff value (B3, B7) displayed significant difference (P <0.05) in various intestinal segments, The Ka value and Peff value of PRS was different from each other with the highest absorption in duodenum (duodenum > jejunum > colon > ileum); The PRS displayed excessive satuation as the concentration increased over 0.05-2.5 g · L(-1). There were no obvious linear correlations between Peff values and concentrations in duodenum (0.6007 ≤ R2 ≤ 0.7727); Ka and Peff value declined when the PRS was perfused with P-glycoprotein promoter digoxin, on the other hand, inclined when perfused with P-glycoprotein inhibitor verapamil with significant difference among PRS B3, BD, B7, B11 (P <0.05). All the above results demonstrated that B3, BD, B7 were greatly influenced by absorption sites, duodenum was the main absorption site; PRS didn't entirely transported in a concentration dependent manner, and the transporter-protein involved the transportation, so the intestinal absorption of the five pulchinenosides was not entirely passive diffusion; and PRS might be the substrates of P-glycoprotein.
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Absorción Intestinal , Ácido Oleanólico/farmacocinética , Pulsatilla/química , Saponinas/farmacocinética , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/fisiología , Animales , Masculino , Ratas , Ratas WistarRESUMEN
OBJECTIVE: To elucidate the role of mast cells (MC) activation in the jejunal mucous membrane in the pathogenesis of cryptosporidiosis (CPS) and explore the mechanism of prevention and treatment of radix sophorae flavescetis(RSF) mixture on CPS. METHODS: A total of 30 healthy male BALB/c mice were randomly divided into a normal control group, CPS model control group and RSF mixture experimental group. The mice of CPS model were inoculated intragastrically with 1 x 10(5) Cryptosporidium oocyst (CSO). The mice in the RSF mixture experimental group were treated with inoculation of RSF mixture (0.2 ml doses) twice one week for three weeks continuously after CPS models were established. Pathological changes of the jejunal mucosa membrane were observed by a light microscope. The MCs were stained by toluidine blue, the number of mast cells was recorded and the changes of degranulation were observed. RESULTS: The HE staining showed inflammatory pathological changes in the jejunal mucosa membrane of the CPS model control group. After three-week treatment of RSF mixture, the small intestine epithelium was integrated on the whole. The toluidine blue stain showed the number of mast cell in submucosa and muscular layer of the jejunal mucous membrane increased significantly in the model control group (12.80 +/- 0.84) compared with those of the normal control group (1.60 +/- 0.89) (P < 0.01) and an obvious degranulation was seen in the CPS model control group. The number of mast cells of the mice in the RSF mixture experimental group decreased significantly (P < 0.01) and the number (2.00 +/- 0.71) and morphous were closed to the normal after administration for three weeks. CONCLUSIONS: MC activation is involved in the intestinal inflammatory response caused by Cryptosporidium. RSF mixture could decline the number of MC, inhibit the activation and degranulation of MC in the jejunal mucosa membrane of CPS mice to reduce inflammation and repair the damaged intestinal mucosa, which may realize the purpose of treatment of CPS.
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Criptosporidiosis/tratamiento farmacológico , Criptosporidiosis/inmunología , Cryptosporidium/fisiología , Medicamentos Herbarios Chinos/administración & dosificación , Mucosa Intestinal/inmunología , Yeyuno/inmunología , Mastocitos/inmunología , Animales , Criptosporidiosis/parasitología , Cryptosporidium/efectos de los fármacos , Cryptosporidium/aislamiento & purificación , Humanos , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/fisiología , Yeyuno/efectos de los fármacos , Yeyuno/parasitología , Masculino , Mastocitos/efectos de los fármacos , Ratones , Ratones Endogámicos BALB CRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Fritillaria thunbergii Miq. has been traditionally used in China as antitussive and expectorant herbs, and newly used in the clinical treatment of leukemia in recent years. AIM: To investigate whether gender exerted a significant influence on the pharmacokinetics, tissue distribution and excretion of peimine and peiminine in Sprague-Dawley rats who were given a single oral administration of 4.25 g/kg Fritillaria thunbergii Miq. extract. MATERIALS AND METHODS: Sprague-Dawley rats were assigned into two groups based on the gender and orally administered 4.25 g/kg Fritillaria thunbergii Miq. extract for each individual pharmacokinetics, tissue distribution and excretion study. RESULTS: Compared with female rats, peimine and peiminine were eliminated slowly from male rat plasma, and significant gender-related differences were observed in the pharmacokinetic parameters. Drug blood and tissue levels in male rats were significantly higher than the female counterparts except for several tissues, such as fat, muscle and skin. Gender also exerted a significant influence on the urine excretion but such effect was not observed in the feces excretion study. CONCLUSIONS: Gender exerted a significant influence on the pharmacokinetics, tissue distribution and urine excretion of peimine and peiminine. It is assumed that the sex-associated differences of peimine and peiminine in rats might be mainly result from sex-dependent expression and activity of drug metabolism enzymes and P-glycoprotein.
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Cevanas/farmacocinética , Cromatografía Liquida/métodos , Extractos Vegetales/farmacocinética , Caracteres Sexuales , Espectrometría de Masas en Tándem/métodos , Animales , Femenino , Fritillaria/química , Masculino , Extractos Vegetales/química , Ratas , Ratas Sprague-Dawley , Distribución TisularRESUMEN
To study the in situ intestinal absorption kinetics and compatibility influence of peimine and peiminine in rats, the absorption of peimine and peiminine in small intestine (duodenum, jejunum and ileum) and colon of rats was investigated using in situ single-pass perfusion method and the drug content was measured by HPLC-ELSD. Perfusion rate, pH, concentration of drug, gender and bile duct ligation can significantly affect the absorption of peimine and peiminine, the Ka, and Papp values in the condition of pH 6.8 and pH 7.4 had significant difference (P<0.01), as drug concentration irlcreased, the absorption parameters of peimine and peiminine decreased, Ka and Papp between low concentrations and middle concentrations was significant difference (P<0.01). Verapamil can not affect Ka and Papp of peimine and peiminine which are in the extract (P> 0.05). Bitter almonds and licorice can significantly reduce the absorption of peimine and peiminine with the usual dose (P<0.01), extracted separately and together had no significant difference on Ka and Papp (P> 0.05). Experimental results show that the absorption features of peimine and peiminine are basically the same, both of them could be absorbed at all segments of the intestine in rats and had no special absorption window, and with significant differences between male and female individuals. The absorption of peimine and peiminine complies with the active transport and facilitated diffusion in the general intestinal segments. Bitter almond and licorice can reduce the intestinal absorption rate ofpeimine and peiminine.
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Cevanas/farmacocinética , Fritillaria/química , Absorción Intestinal , Animales , Cevanas/administración & dosificación , Cevanas/aislamiento & purificación , Colon/metabolismo , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Femenino , Glycyrrhiza/química , Ácido Glicirrínico/aislamiento & purificación , Ácido Glicirrínico/farmacología , Absorción Intestinal/efectos de los fármacos , Intestino Delgado/metabolismo , Masculino , Perfusión , Raíces de Plantas/química , Plantas Medicinales/química , Prunus dulcis/química , Ratas , Ratas Sprague-Dawley , Factores SexualesRESUMEN
OBJECTIVE: To investigate the protective effect of radix sophorae flavescentis (RSF) mixture on intestinal mucosa in mice infected with Cryptosporidium parvum. METHODS: Thirty BALB/c male mice were randomly divided into control group, infection group and RSF mixture treatment group. Mice of the posterior two groups were inoculated intragastrically with 1 x 10(5) C. parvum oocysts, immunosuppressed with dexamethasone (5 microg/ml) and gentamycin sulfate (40 microg/ml) in drinking water. At the 8th day post-infection, mice in RSF mixture treatment group were treated with 0.2 ml dose of RSF mixture twice a week (three-day intervals) for three weeks. The mice in infection group and RSF mixture treatment group were monitored for oocyst shedding in fecal pellets every two days after treatment. At 28 days after infection, experimental mice were sacrificed, jejunal tissue was removed for preparation of paraffin-embedded sections. The changes of CD3+, CD4+, CD8+ T lymphocytes and IgA plasmocytes in intestinal mucosa were determined by immunohistochemistry. In addition, jejunum of infected mice and treated mice were collected, and ultrastructural changes were observed under electron microscopy. RESULTS: Compared with infection group, the level of oocyst shedding was obviously lower and the time of the oocyst discharging was significantly shorter in RSF mixture treatment group. The proportion of CD3+, CD4+ T lymphocyte and CD4+/CD8+ T cell ratio in infection group (49.7% +/- 2.4%, 25.7% +/- 2.2%, 1.1 +/- 0.3) were significantly lower than that of treatment group (62.4% +/- 1.4%, 37.5% +/- 3.1%, 1.5 +/- 0.3) and control group (66.5% +/- 1.9%, 40.1% +/- 1.8%, 1.5 +/- 0.2) (P < 0.01). CD8+ T lymphocytes showed no significant difference in each group (P > 0.05). The number of IgA plasmocytes in treatment group (52.7 +/- 3.5) was significantly higher than that of control group (8.3 +/- 2.3) and infection group (33.7 +/- 2.6) (P < 0.01). After administration for three weeks, the damaged C. parvum parasites were seldom seen in mouse jejunum, and lysosomes appeared in large number, RSF mixture treatment improved mitochondrial structure and repaired microvilli. In infection group, mitochondria ridges were significantly broken and microvilli surrounding C. parvum oocysts were shed, resulting in the appearance of crater-like lesions on the surface, the oocyst wall and host cell membrane fused together. CONCLUSION: RSF mixture is effective against Cryptosporidium parvum. The damage of intestinal mucosa in infected mice can be repaired after treatment.
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Criptosporidiosis/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Mucosa Intestinal/efectos de los fármacos , Animales , Cryptosporidium parvum , Medicamentos Herbarios Chinos/uso terapéutico , Mucosa Intestinal/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Sophora/químicaRESUMEN
OBJECTIVE: To investigate the effect of Toxoplasma gondii infection on the proliferation, differentiation and migration of the embryonic neural stem cells (NSCs) in early pregnancy of rat. METHODS: Twelve pregnant Sprague-Dawley rats were randomly divided into control and infection groups. Rats in the infection group were each inoculated intraperitoneally with 1 x 10(5) T. gondii RH strain tachyzoites at day 1 (E1 day). Same amount of physiological saline was intraperitoneally injected for rats in control group. At E5 day, blood samples were taken from caudal vein and Giemsa staining of blood cells was performed to find T. gondii. At E9, E10 and E11 day, two rats in each group per time point were sacrificed and reverse transcription PCR (RT-PCR) was performed to detect B1 gene expression of T. gondii in amniotic fluid to confirm T. gondii infection. NSCs were cultured in vitro. The proliferation level was detected by methyl thiazolyl tetrazolium (MTT) assay. After differentiation culture of NSCs, the immunofluorescence assay was conducted to detect the expression of nestin, microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) to calculate the ratio of NSCs which differentiated to neurons and astrocytes. The embryonic nerve tissues at E9, E10 and E11 day in each group were taken to make frozen sections. The immunofluorescence assay was carried out to detect the expression of neuronal cell adhesion molecule (NCAM) in the nerve tissues at different developmental stages. RESULTS: Both the results of blood smears and RT-PCR confirmed that the pregnant rats and embryos were all infected by T. gondii in infection group. The morphology of the cultured NSCs under microscope was consistent with the characteristics of the normal NSCs. In addition, the NSC biomarker nestin protein was stained positive. The MTT assay showed that the proliferation level was lower in infection group than that of the control, and statistical differences were found between the two groups at day 3 and 4 after passages (P < 0.05). The immunofluorescence staining of MAP2 and GFAP showed that the percentage of neuron differentiation was 15.15% (55/363) in control group and 8.73% (31/355) in infection group, respectively, with a statistical difference (P < 0.05), and the percentage of astrocyte differentiation was 53.35% (199/374) and 67.48% 249/369), respectively (P > 0.05). In both groups, NCAM protein was found expressed at E9, E10 and E11 day in embryo nerve tissues. The fluorescence became stronger with time. The expression level in control group was significantly higher than that in infection group (P < 0.01). CONCLUSION: T. gondii infection at early gestation may inhibit the proliferation, differentiation and migration of neural stem cells in rats.
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Células Madre Embrionarias/citología , Células-Madre Neurales/citología , Complicaciones Infecciosas del Embarazo/parasitología , Toxoplasmosis/patología , Animales , Diferenciación Celular , Proliferación Celular , Células Madre Embrionarias/parasitología , Femenino , Células-Madre Neurales/parasitología , Neuronas/citología , Embarazo , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To study the optimum inclusion process of clolloid mill for borneolumsyntheticum from Huoxuezhitong Capsules. METHODS: The study was carried out with orthogonal design. The borneol content in inclusion complex, the utilization ratio of borneol, and synthesis grading were used as evaluating indicator to review the effects of 3 factors, including the proportion of borneolumsyntheticum to beta-cyclodextrin (mol: mol), milling time and water addition. RESULTS: The optimum process were borneolumsyntheticum: beta-cyclodextrin was 1:1, the milling time was 40 min, water addition was double. CONCLUSION: The optimum process is feasible, and fit for industrial production, with the advantage of high utilization ratio, shot time and low energy consumption.
Asunto(s)
Canfanos/análisis , Medicamentos Herbarios Chinos/química , Aceites Volátiles/análisis , Tecnología Farmacéutica/métodos , beta-Ciclodextrinas , Disponibilidad Biológica , Cápsulas , Cromatografía de Gases , Cromatografía en Capa Delgada , Portadores de Fármacos , Estabilidad de Medicamentos , Medicamentos Herbarios Chinos/análisis , Aceites Volátiles/química , Factores de Tiempo , Agua/administración & dosificaciónRESUMEN
OBJECTIVE: To study optimum inclusion process condition for volatile oil with beta-cyclodextrin (beta-CD). METHODS: The optimum preparation condition were investigated by orthogonal design. The influence of utilization beta-CD and oil, inclusion temperature and time were studied to determin the utilization ratio of volatile oil and oil-bearing rate. RESULTS: The optimum preparation conditions were established as: volatile oil: beta-CD was 1:10, the inclusion temperature was 60 degrees C and inclusion time was 3h. CONCLUSION: The method is convenient and can he used for mass production.