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Objective To understand the characteristics and developmental differences between cerebral organoids in vitro and normal cerebral cortices in vivo. Methods 1. Grouping: cerebral cortices in vivo group and cultured cerebral organoids in vitro group. 2. Sample collection: cortical tissues were collected from Kunming mouse embryos at embryonic day 7.5(E7.5), E9.5, E11.5, E14.5, and postnatal day 3 (P3) or P7. Three specimens were taken from each group. Meanwhile, cerebral organoids were cultured with mouse induced pluripotent stem cells (iPSCs), and samples at different culture time point were collected, and more than 3 samples were collected at each time point. 3. Detection method: the distribution of different types of cells in each group of specimens was analyzed by immunofluorescent staining. Results While relative similarities between in vivo cerebral cortical development and the cerebral organoids in vitro were observed, including the histogenesis, and the morphological differentiation of nerve cells and glial cells, the lamellar architecture of cerebral cortex in mouse brain was not observed in cerebral organoids. Conclusion The development of cerebral organoids in vitro has some similarity with body's cortical development. Therefore, cerebral organoids can be used to a substitution of cortex and diseases' models, but improvement of the existing technologies is necessary.
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Osteoarthritis (OA) is one of the most common joint degenerative diseases in the world. At present, the management of OA depends on the lifestyle modification and joint replacement surgery, with the lifespan of prosthesis quite limited yet. Effective drug treatment of OA is essential. However, the current drugs, such as the non-steroidal anti-inflammatory drugs and acetaminophen, as well as glucosamine, chondroitin sulfate, hyaluronic acid, are accompanied by obvious side effects, with the therapeutic efficacy to be enhanced. Recently, novel reagents such as IL-1 antagonists and nerve growth factor inhibitors have entered clinical trials. Moreover, increasing evidence demonstrated that active ingredients of natural plants have great potential for treating OA. Meanwhile, the use of novel drug delivery strategies may overcome the shortcomings of conventional preparations and enhance the bioavailability of drugs, as well as decrease the side effects significantly. This review therefore summarizes the pathological mechanisms, management strategies, and research progress in the drug molecules including the newly identified active ingredient derived from medicinal plants for OA therapy, with the drug delivery technologies also summarized, with the expectation to provide the summary and outlook for developing the next generation of drugs and preparations for OA therapy.
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Objective To investigate the effects of the development of Purkinje cells and expression of cerebellin in postnatal mice with intrauterine hypoxia. Methods Twenty healthy adult Kunming mice were randomly divided into two groups : control group and hypoxia group, with 10 mice in each group. Mice in the hypoxia group were placed in the hypoxia chamber of the animal since the 14th day of gestation to make an animal model of intrauterine hypoxia. After the mother gave birth, the experimental animals were divided into hypoxia group and control group. There were 6 age groups including postnatal day (P)0, P5, P9, P14, P21 and P30 in each group, and 5 mice in each age group. The cerebellum tissue was taken for vibrating sectioning. The developmental changes of calbindin-positive Purkinje cells were detected by immunofluorescence technique. The expression of cerebellin in Purkinje cell protuberances was detected by cerebellin (CBLN)l, CBLN4 and calbindin double labeling. Finally, Western blotting was used to semi-quantitatively analyze the protein expression of cerebellar peptide in cerebellum at each time point. Results Compared with the control group of the same age, the number of cerebellar Purkinje cells in the hypoxic group decreased, the dendritic branches decreased, and the arrangement was disordered, and the expression of CBLN1 and CBLN4 in the cortex were significantly reduced. Conclusion Intrauterine hypoxia leads to abnormal development of the cerebellar Purkinje cells and synaptic changes.
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The purpose of the study was to investigate the effect of sophoridine on the proliferation and apoptosis of human gastric cancer MKN45 cells and the possible mechanism. MKN45 cells were randomly divided into control and sophoridine (including 6 subgroups) groups. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) colorimetric method. The protein expression of high mobility group-box 3 (HMGB3) was observed by immunocytochemical staining and Western blot. Hoechst 33342 staining method was used to observe the morphological changes of cells treated with sophoridine. Apoptosis was detected by flow cytometry. The results showed that the proliferation of cells was inhibited by 48-hour treatment of sophoridine in a dose-dependent manner. Compared with control group, sophoridine group showed decreased HMGB3 protein expression and increased apoptotic rate. These results suggest that sophoridine can inhibit the proliferation of MKN45 cells and promote their apoptosis, which may be related to down-regulation of HMGB3 protein expression.
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Orally disintegrating tablets (ODT), a kind of new solid tablet that rapidly disintegrates to work in the mouth, has became the hot form of new drug research in recent years with many advantages, such as the convenient taking, a widely applicable people, fast acting, high bioavailability, good compliance, and so on. ODT has been widely used in chemical medicines, while the application of it in traditional Chinese medicines (TCMs) is still in the stage of development The development of TCMs ODT provides a new direction for the research of Chinese medicine new dosage, accelerates the pace of connecting to the world and modernization of Chinese medicine. This dosage has a broad market prospect, and its quality control and assessment standards, taste, the disintegration time in vitro and evaluation method are the key factors that affect the industrialization, standardization of Chinese medicine ODT. Therefore, this paper reviewed the characteristics, preparation, taste masking technology and quality evaluation with new technology of ODT. Meantime, numerous application examples of ODT used in traditional Chinese medicine were described. We expect to provide the reference and utilization for the development of traditional Chinese medicine orally disinteeratine tablets.
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Humanos , Administración Oral , Composición de Medicamentos , Métodos , Medicina Tradicional China , Solubilidad , Comprimidos , Química , GustoRESUMEN
The aim of the study was to investigate the effect of astragalus on differentiation of human amniotic epithelial cell line WISH into neurons, the expression of Notch1 gene and cell viability. WISH were randomly divided into astragalus group (4 subgroups), alltransretinoic acid (RA) group and control group. Astragalus group and RA group were induced to differentiate into neurocytes by using chemical inducer RA and astragalus, respectively. The expression of neuron-specific enolase (NSE), microtubule associated protein 2 (MAP-2), Nestin and GFAP of induced cells in three groups were detected using immunocytochemical method. RT- PCR was further used to detect the expression of Oct4, Notch1, Hes1, Nestin and NSE. The cell viability was measured by methyl thiazolyl tetrazolium methods. Under the convert microscope it was observed that WISH cells started to change their shape, and there were several axon or dendrite-like processes out from the cell body induced by astragalus for 24 h or RA for 12 h. The positive cell rates of NSE and MAP-2 in 100 μL/mL astragalus-induced group were less than those in RA-induced group at 48 h (P < 0.05), but higher than those in control group. Cell viability in astragalus group was higher than that of RA group (P < 0.05). While the positive cell rates of Nestin and GFAP in 100 μL/mL astragalus-induced group were higher than those in RA-induced group at 48 h (P < 0.05). The positive cell rates of Nestin in the two induced groups were lower than those in control group. RT-PCR showed that the expressions of Oct4, Notch1 and Hes1 in RA and astragalus (100 μL/mL) groups were less than those in control group, but the expression of NSE was higher than that in control group. These results suggest that astragalus (especially at 100 μL/mL, 48 h) and RA can both induce human amniotic epithelial cell line WISH cells into neuron-like cells, but astragalus induction has a higher cell survival rate than RA induction, and the expression of Notch1 signal molecules is inhibited during the induction.
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Humanos , Amnios , Biología Celular , Planta del Astrágalo , Química , Diferenciación Celular , Línea Celular , Supervivencia Celular , Células Epiteliales , Biología Celular , Neuronas , Biología Celular , Receptor Notch1 , Metabolismo , Tretinoina , FarmacologíaRESUMEN
<p><b>BACKGROUND</b>Deep venous thrombosis (DVT) can result in pulmonary embolism, a fatal complication that is due to the dislodgement and movement of a blood clot (thrombus) from a limb into the lungs. Genetic risk factors related to DVT development include mutations in coagulation proteins, especially the endothelial protein C receptor (EPCR), a component of the anticoagulation protein C (PC) pathway. The objective of the present study was to analyze the relationship between the 6936A/G polymorphism in the EPCR gene and the occurrence of DVT.</p><p><b>METHODS</b>This study involved 65 patients with DVT and 71 age- and gender-matched healthy controls. Peripheral blood samples were collected from all subjects. Plasma levels of soluble EPCR (sEPCR) were measured by enzyme-linked immunosorbent assay. Genomic DNA was extracted and EPCR gene product was amplified by a standard PCR reaction. Gene product bands were sequenced to identify EPCR gene polymorphisms.</p><p><b>RESULTS</b>In the control group, the level of sEPCR in subjects with 6936AG genotype was significantly higher than that in subjects with 6936AA genotype ((0.97 ± 0.32) pg/ml vs. (0.61 ± 0.24) pg/ml, P < 0.01). Similarly in the DVT group, the level of sEPCR in subjects with the 6936AG were greater than that in subjects with the 6936AA genotype ((0.87 ± 0.21) pg/ml vs. (0.50 ± 0.18) pg/ml, P < 0.01). The sEPCR level in DVT patients was significantly higher than that in healthy controls ((0.68 ± 0.32) pg/ml vs. (0.54 ± 0.22) pg/ml, P < 0.05). The 6936AG genotype frequency in DVT patients was significantly higher than that in healthy controls (P < 0.05). In contrast, the 6936AA genotype frequency in DVT patients was lower than that in healthy controls (P < 0.05). Subjects carrying 6936AG had an increased risk of thrombosis (OR = 2.75, 95%CI: 1.04 - 7.30, P < 0.05).</p><p><b>CONCLUSIONS</b>EPCR gene 6936A/G polymorphism is associated with increased plasma levels of sEPCR. Subjects carrying 6936AG likely have an increased risk of thrombosis.</p>
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Femenino , Humanos , Masculino , Persona de Mediana Edad , Factores de Coagulación Sanguínea , Genética , Estudios de Casos y Controles , Predisposición Genética a la Enfermedad , Genética , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Genética , Receptores de Superficie Celular , Genética , Trombosis de la Vena , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To investigate the relationship between endothelial protein C receptor(EPCR) gene 6936A/G polymorphism and deep vein thrombosis (DVT).</p><p><b>METHODS</b>The study group included 65 DVT patients and 71 normal controls. Plasma sEPCR was measured by ELISA. Genomic DNA was extracted by using Genomic Purification Kit. A 315bp EPCR product was amplified by a standard PCR reaction, and the bands were confirmed by direct sequencing after purification.</p><p><b>RESULTS</b>(1) sEPCR levels in healthy controls with 6936AG genotype were significantly higher than that in those with 6936AA genotype \[(0.97 ± 0.32) ng/L vs (0.61 ± 0.24) ng/L, P < 0.01)\], and so did in DVT patients \[(0.87 ± 0.21) ng/L vs (0.50 ± 0.18) ng/L, P < 0.01\]. (2) The sEPCR levels of DVT patients \[(0.68 ± 0.32) ng/L\] were significantly higher than that of healthy controls \[(0.54 ± 0.22) ng/L\](P < 0.05). (3) The distribution of 6936A/G genotype was higher in DVT patients than in healthy controls (P < 0.05). (4) Subjects with 6936A/G had an increased risk of thrombosis (OR = 2.75, 95%CI = 1.04 - 7.30) (P < 0.05).</p><p><b>CONCLUSIONS</b>EPCR gene 6936A/G polymorphism is associated with increased plasma sEPCR levels. The sEPCR levels in DVT patients were significantly higher than that in healthy controls. The subject with 6936AG likely had an increased risk of thrombosis.</p>
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Humanos , Estudios de Casos y Controles , Polimorfismo Genético , Proteína C , Metabolismo , Trombosis , Trombosis de la Vena , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To explore whether transplantation of autologous bone marrow stem cells might augment angiogenesis and collateral vessel formation in a canine model of hindlimb ischemia.</p><p><b>METHODS</b>CD34(+) stem cells were centrifugation through Ficoll and an immune magnetic cell sorting system from bone marrow (20 ml) of canine (n = 5) and induced into endothelial cells with VEGF in vitro, and expression of von Willebrand factor. Bilateral hindlimb ischemia was surgically induced in canines and Dil fluorescence labeled autologous stem cells were transplanted into the ischemic tissues.</p><p><b>RESULTS</b>Four weeks after transplantation, fluorescence microscopy revealed that transplanted cells were incorporated into the capillary network among preserved skeletal myocytes. The stem cells transplanted group had more angiographically detectable collateral vessels, a higher capillary density (12.0 +/- 2.8 vs. 5.0 +/- 1.6 per field; t = 4.17 P < 0.05) and a higher ABI (0.58 +/- 0.14 vs. 0.32 +/- 0.11; t = 2.95, P < 0.05).</p><p><b>CONCLUSIONS</b>Direct local transplantation of autologous bone marrow CD34(+) stem cells seems to be a useful strategy for therapeutic neovascularization in ischemic tissues in adults, consistent with "therapeutic vasculogenesis."</p>
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Animales , Perros , Femenino , Masculino , Antígenos CD34 , Diferenciación Celular , Modelos Animales de Enfermedad , Células Endoteliales , Biología Celular , Trasplante de Células Madre Hematopoyéticas , Métodos , Células Madre Hematopoyéticas , Química , Biología Celular , Fisiología , Miembro Posterior , Isquemia , Terapéutica , Neovascularización FisiológicaRESUMEN
<p><b>OBJECTIVE</b>To exploration the endothelialization of prostheses with bone marrow CD(34)(+) cells.</p><p><b>METHODS</b>CD(34)(+) cells were isolated from bone marrow of carine by an immune magnetic cell sorting system and induced into endothelial cells with VEGF. Seeding the cells to PTFE prostheses which implanted the abdominal aorta artery (AAA) and inferior vena cava (IVC).</p><p><b>RESULTS</b>The isolated cells from bone marrow were CD(34)(+) by flow cytometer which could differentiate into endothelial cells in vascular endothelial growth factor (VEGF). The endothelial cells were identified by immunostaining and transmission electron microscope. The obliteration rate of the seeding grafts implanting AAA was 0%, the stenosis rate 12.5%; the obliteration rate implanting IVC 12.5%, the stenosis rate 25%.</p><p><b>CONCLUSION</b>CD(34)(+) cells were isolated from bone marrow by an immune magnetic cell sorting system and were able to be induced into endothelial cells with VEGF in vitro. PTFE prostheses seeding CD(34)(+) cells have the ideal endothelialization and patency.</p>
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Animales , Perros , Femenino , Masculino , Antígenos CD34 , Metabolismo , Prótesis Vascular , Implantación de Prótesis Vascular , Médula Ósea , Células de la Médula Ósea , Metabolismo , Diferenciación Celular , Células Endoteliales , Citometría de Flujo , Microscopía Electrónica , Factor A de Crecimiento Endotelial Vascular , FarmacologíaRESUMEN
<p><b>OBJECTIVE</b>To offer evidences for quality control of medicinal Asteropyrum plants.</p><p><b>METHOD</b>Pharmacognostic studies were made through field collection, market investigation, document utilization, comparative morphology and histology.</p><p><b>RESULT</b>The shape and properties, microscopic characteristics in root, rhizome, leaves were worked out.</p><p><b>CONCLUSION</b>Asteropyrum can be distinguished from close relative Coptis plants by microscopic and histology characteristics. Two species in Asteropyrum can also be identified by microscopic and histology characteristics, and the morphological and histology characteristics can be used as evidences for quality control of medicinal Asteropyrum plants.</p>
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Coptis , Farmacognosia , Hojas de la Planta , Química , Raíces de Plantas , Química , Plantas Medicinales , Control de Calidad , Ranunculaceae , Especificidad de la EspecieRESUMEN
Objective To explore the effect of intrauterin hypoxia on proliferation of neural stem cells(NSCs)and memory and learning capacity of juvenile rats.Methods Conceiving female SD rats(n=20)were divided randomly into 2 groups:control group(n=10)and hypoxia group(n=10).The rats of hypoxia group were put into 3 gases incubator to make the injured brain newborn rats model,rats of control group were not done so.The juvenile rats were taken to do the experiment of water maze after they were born in 30 d,then killed when the experiment was over,the brains of the rats were taken,immobilization,paraffin imbedding,slicing(hippocampus),common immunohistochemistry staining were used to detect the Nestin positive NSCs.Results Optical density of Nestin positive cells in hippocampus dentation of hypoxia group(0.16?0.10)was higher than that of control group(0.15?0.09)(t=3.24 P