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1.
Chinese Pharmacological Bulletin ; (12): 162-170, 2024.
Article in Chinese | WPRIM | ID: wpr-1013611

ABSTRACT

Aim To investigate the targeting mechanism of miR-23b on PINKl/Parkin pathway in transdifferentiation of NRK-52E cellsinduced by TGF-β1, and to elucidate the intervention mechanism of Qingshen granules drug-containing serum on NRK-52E cell transdifferentiation. Methods Ultra-high performance liquid chromatography ( UPLC ) fingerprinting method was used to analyze Qingshen granules. The NRK-52E transdifferentiation model induced by TGF-β1 was constructed. The NRK-52E cells were divided into simulated no-load control group, miR-23b-5p simulated group, inhibitor no-load control group, and miR-23b-5p inhibitor group, after transfection with siRNA, and the effect of miR-23b-5p on PINK1 expression was ob-served. The NRK-52E cells were then divided into normal group, TGF-(31 group, Qingshen granule group, miR-23 b-mimic group, miR-23 b-mimic group, and miR-23b-mimic + Qingshen granule group. Western blot was used to detect the expression of Pinkl, Parkin, LC3 n, Beclin-1, P62 and a-SMA proteins, and RT- PCR was used to detect the expression of miR-23 b-5p, Pinkl, Parkin, Beclin-1 and a-SMA mRNA in NRK- 52E cells. Dual-Luciferase Reporter gene experiment was used to detect the targeting relationship between miR-23b-5p and PINKL Results UPLC fingerprinting method found 11 active components in Qingshen granules. After overexpression of miR-23b-5p, the expression of PINkl mRNA significantly increased (P 0. 05 ). The experimental results showed that the expressions of miR- 23b-5p, Pinkl, Parkin, Beclin-1, LC3 II and LC3 II/ I ratio in TGF-β1 group were significantly lower than those in normal group, but the expressions of P62 and a-SMA were significantly higher than those in normal group ( P <0.05). The expressions of miR-23 b-5 p, Pinkl, Parkin, Beclin-1, LC3 II and LC3 11/ I ratio in Qingshen granule group and miR-23 b-mimic group were significantly higher than those in TGF-β1 group, and the expressions of P62 and a-SMA were significantly lower than those in TGF-β1 group (P < 0. 05 ). The performance of miR-23 b-mimic + Qingshen granule group was better than that of miR-23 b-mimic group (P < 0. 05 ). Conclusions Qingshen granules can up- regulate the expression of miR-23b-5p in NRK-52E cellsand inhibit the transdifferentiation process of NRK- 52E cells by enhancing the mitochondrial autophagy activity mediated by PINKl/Parkin pathway.

2.
Acta Pharmaceutica Sinica ; (12): 2155-2167, 2023.
Article in Chinese | WPRIM | ID: wpr-999108

ABSTRACT

The medicinal history of Pien Tze Huang is long, and it is the only "double top secret" variety of technology and formula at present. It has the effects of clearing heat and detoxifying, detumescence and pain, cooling blood and removing blood stasis. At present, researchers have analyzed and identified some compounds in Pien Tze Huang and its precious medicinal materials, such as Panax notoginseng, calculus bovis, snake gall and musk, and conducted activity screening, pharmacokinetics and pharmacological related studies on these chemical components. It was found that Pien Tze Huang had a significant effect on the treatment of acute and chronic hepatitis, ulcer, colon cancer, liver cancer and other diseases. The purpose of this paper is to systematically discuss the research achievements of researchers in recent years on the material basis, pharmacological effects and clinical application of Pien Tze Huang, with a view to providing ideas for the further research of Pien Tze Huang.

3.
Journal of Experimental Hematology ; (6): 364-376, 2023.
Article in Chinese | WPRIM | ID: wpr-982068

ABSTRACT

OBJECTIVE@#To investigate and analyze the effect of CXC chemokine receptor 1/2 (CXCR1/2) targeting inhibitor Reparixin combined with cytarabine (Ara-C) on the malignant biological behaviors of acute myeloid leukemia cells and its effect on the expression of the CXCR family, while exploring the accompanying molecular mechanism, providing scientific basis and reference for new molecular markers and targeted therapy for AML.@*METHODS@#Acute myeloid leukemia U937 cells were treated with different concentrations of Reparixin, Ara-C alone or in combination, and the cell morphology was observed under an inverted microscope; Wright-Giemsa staining was used to detect cell morphological changes; CCK-8 method was used to detect cell proliferation; the ability of cell invasion was detected by Transwell chamber method; the ability of colony formation was detected by colony formation assay; cell apoptosis was detected by Hoechst 33258 fluorescent staining and Annexin V/PI double-staining flow cytometry; monodansylcadaverine(MDC) staining was used to detect cell autophagy; the expression of apoptosis, autophagy and related signaling pathway proteins was detected by Western blot and the expression changes of CXCR family were detected by real-time quantitative polymerase chain reaction (qRT-PCR).@*RESULTS@#Reparixin could inhibit the proliferation, invasion, migration and clone formation ability of U937 cells. Compared with the single drug group, when U937 cells were intervened by Reparixin combined with Ara-C, the malignant biological behaviors such as proliferation, invasion and colony formation were significantly decreased, and the levels of apoptosis and autophagy were significantly increased (P<0.01). After Reparixin combined with Ara-C intervenes in U937 cells, it can up-regulate the expression of the pro-apoptotic protein Bax and significantly down-regulate the expression of the anti-apoptotic protein Bcl-2, and also hydrolyze and activate Caspase-3, thereby inducing cell apoptosis. Reparixin combined with Ara-C could up-regulate the expressions of LC3Ⅱ and Beclin-1 proteins in U937 cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (P<0.01). MDC result showed that the green granules of vesicles increased significantly, and a large number of broken cells were seen (P<0.01). Reparixin combined with Ara-C can significantly inhibit the phosphorylation level of PI3K, AKT and NF-κB signaling molecule, inhibit the malignant biological behavior of cells by inhibiting the activation of PI3K/AKT/NF-κB pathway, and induce programmed cell death. Ara-C intervention in U937 cells had no effect on the expression of CXCR family (P>0.05). The expression of CXCR1, CXCR2, and CXCR4 mRNA could be down-regulated by Reparixin single-agent intervention in U937 cells (P<0.05), and the expression of CXCR2 was more significantly down-regulated than the control group and other CXCRs (P<0.01). When Reparixin and Ara-C intervened in combination, the down-regulated levels of CXCR1 and CXCR2 were more significant than those in the single-drug group (P<0.01), while the relative expressions of CXCR4 and CXCR7 mRNA had no significant difference compared with the single-drug group (P>0.05).@*CONCLUSION@#Reparixin combined with Ara-C can synergistically inhibit the malignant biological behaviors of U937 cells such as proliferation, invasion, migration and clone formation, and induce autophagy and apoptosis. The mechanism may be related to affecting the proteins expression of Bcl-2 family and down-regulating the proteins expression of CXCR family, while inhibiting the PI3K/AKT/NF-κB signaling pathway.


Subject(s)
Humans , U937 Cells , Cytarabine/therapeutic use , Receptors, Interleukin-8A , NF-kappa B , Proto-Oncogene Proteins c-akt , Phosphatidylinositol 3-Kinases , Leukemia, Myeloid, Acute/genetics , Apoptosis , Cell Proliferation , Apoptosis Regulatory Proteins , Proto-Oncogene Proteins c-bcl-2 , RNA, Messenger , Cell Line, Tumor
4.
Journal of Experimental Hematology ; (6): 1394-1402, 2023.
Article in Chinese | WPRIM | ID: wpr-1009994

ABSTRACT

OBJECTIVE@#To analyze the effects of mangiferin combined with bortezomib on the proliferation, invasion, apoptosis and autophagy of human Burkitt lymphoma Raji cells, as well as the expression of CXC chemokine receptors (CXCRs) family, and explore the molecular mechanism between them to provide scientific basis for basic research and clinical work of Burkitt lymphoma.@*METHODS@#Raji cells were intervened with different concentrations of mangiferin and bortezomib alone or in combination, then cell proliferation was detected by CCK-8 assay, cell invasion ability was detected by Transwell chamber method, cell apoptosis was detected by Annexin V/PI double-staining flow cytometry, apoptosis, autophagy and Akt/mTOR pathway protein expression were detected by Western blot, and the expression changes of CXCR family was detected by real-time quantitative PCR (RT-qPCR).@*RESULTS@#Different concentrations of mangiferin intervened Raji cells for different time could inhibit cell viability in a concentration- and time-dependent manner (r =-0.682, r =-0.836). When Raji cells were intervened by combination of mangiferin and bortezomib, compared with single drug group, the proliferation and invasion abilities were significantly decreased, while the apoptosis level was significantly increased (P <0.01). Mangiferin combined with bortezomib could significantly up-regulate the expression of pro-apoptotic protein Bax and down-regulate the expression of anti-apoptotic protein Bcl-2 after intervention in Raji cells. Caspase-3 was also hydrolyzed and activated, and then induced the apoptosis of Raji cells. Mangiferin combined with bortezomib could up-regulate the expression of LC3Ⅱ protein in Raji cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (P <0.01). Mangiferin combined with bortezomib could significantly inhibit the phosphorylation levels of Akt and mTOR, inhibit the proliferation and invasion of Raji cells by inhibiting Akt/mTOR pathway, and induce cell autophagy and apoptosis. Mangiferin and bortezomib could down-regulate the expressions of CXCR4 and CXCR7 mRNA after single-agent intervention in Raji cells, and the down-regulations of CXCR4 and CXCR7 mRNA expression were more significant when the two drugs were combined (P <0.01). Mangiferin alone or combined with bortezomib had no significant effect on CXCR5 mRNA expression in Raji cells (P >0.05), while the combination of the two drugs could down-regulate the expression of CXCR3 (P <0.05).@*CONCLUSION@#Mangiferin combined with bortezomib can synergistically inhibit the proliferation and invasion of Raji cells, and induce autophagy and apoptosis. The mechanism may be related to the inhibition of Akt/mTOR signaling pathway, down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax, and the inhibition of the expression of CXCR family.


Subject(s)
Humans , Antineoplastic Agents/therapeutic use , Apoptosis/drug effects , Apoptosis Regulatory Proteins/immunology , Autophagy/immunology , bcl-2-Associated X Protein/immunology , Bortezomib/therapeutic use , Burkitt Lymphoma/immunology , Cell Line, Tumor , Cell Proliferation/drug effects , Drug Therapy, Combination , Proto-Oncogene Proteins c-akt , Proto-Oncogene Proteins c-bcl-2 , Receptors, CXCR/immunology , RNA, Messenger , TOR Serine-Threonine Kinases , Xanthones/therapeutic use
5.
Journal of Experimental Hematology ; (6): 794-800, 2023.
Article in Chinese | WPRIM | ID: wpr-982132

ABSTRACT

OBJECTIVE@#To investigate the effect of pure Chinese herbal extract Mangiferin on the malignant biological behaviors of multiple myeloma (MM) cells, and to analyze the molecular mechanism of the anti-myeloma effect of Mangiferin, so as to provide experimental basis for MM replacement therapy.@*METHODS@#U266 and RPMI8226 of human MM cell lines were intervened with different concentrations of Mangiferin. Cell proliferation was detected by CCK-8 method. Annexin V/PI double staining flow cytometry was used to detect cell apoptosis. Western blot was used to detect the expression of apoptosis and related signaling pathway proteins, and real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of matrix metalloproteinase (MMP) and CXC chemokine receptor (CXCR) family.@*RESULTS@#Mangiferin could inhibit the proliferation activity of U266 and RPMI8226 cells and induce cells apoptosis. After Mangiferin intervened in U266, RPMI8226 cells for 48 h, the expression of Bcl-2 family pro-apoptotic protein Bax was up-regulated, while the expression of survivin and Bcl-xL proteins was down-regulated and caspase-3 was hydrolyzed and activated to promote cell apoptosis, besides, the expression of Bcl-2 protein in U266 cells was also significantly down-regulated to induce apoptosis (P<0.05). After Mangiferin intervenes in MM cells, it can not only increase the expression level of tumor suppressor p53, but also induce programmed cell death of MM cells by inhibiting the expression of anti-apoptotic molecules and down-regulating the phosphorylation levels of AKT and NF-κB. In addition, after the intervention of Mangiferin, the expressions of CXCR4, MMP2 and MMP9 in U266 cells were down-regulated (P<0.05), while there is no effect on the expressions of CXCR2, CXCR7 and MMP13 (P>0.05). However, the expressions of CXCR4, MMP9, and MMP13 in RPMI8226 cells were down-regulated (P<0.01), the expression of MMP2 was weakly affected, and the expression of CXCR2 and CXCR7 was basically not affected (P>0.05).@*CONCLUSION@#Mangiferin can inhibit the proliferation and induce apoptosis of MM cells, and its mechanism may be related to inhibiting the activation of NF-κB signaling pathway, affecting the expression of Bcl-2 family proteins, and inhibiting the expression of core members of MMP and CXCR family.


Subject(s)
Humans , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Matrix Metalloproteinase 13 , Cell Line, Tumor , NF-kappa B , Multiple Myeloma/pathology , Cell Proliferation , Apoptosis , Proto-Oncogene Proteins c-bcl-2
6.
Acta Academiae Medicinae Sinicae ; (6): 488-493, 2021.
Article in Chinese | WPRIM | ID: wpr-887884

ABSTRACT

A case of primary oral mucosal diffuse large B-cell lymphoma(DLBCL)due to long-term use of methotrexate(MTX)for the treatment of rheumatoid arthritis(RA)was admitted to the Department of Hematology,Fujian Medical University Union Hospital.We analyzed and discussed the clinical features,diagnosis and treatment,and prognosis of specific malignant lymphoma induced by MTX in this RA patient.Our purpose is to improve the awareness and knowledge of other iatrogenic immunodeficiency-associated lymphoproliferative disorders of clinicians and pathologists.This study provides a new reference for the clinical diagnosis and treatment of MTX-associated DLBCL.


Subject(s)
Humans , Arthritis, Rheumatoid/drug therapy , Lymphoma, Large B-Cell, Diffuse/drug therapy , Lymphoproliferative Disorders , Methotrexate/adverse effects
7.
China Journal of Chinese Materia Medica ; (24): 3144-3154, 2020.
Article in Chinese | WPRIM | ID: wpr-828004

ABSTRACT

To reveal the processing mechanism of Chrysanthemi Flos from the changes of chemical compositions after frying and its effect on the efficacy of liver protection. Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) and ultra high performance liquid chromatography(HPLC) were used for the qualitative and quantitative researches of chemical compositions before and after Chrysanthemi Flos frying. Progenesis QI and SPSS software were used for principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), variable importance projection(VIP) analysis and t-test to identify the compositions with significant changes. Pharmacodynamics experiment was used to investigate the protective effect of crude and fried Chrysanthemi Flos on CCl_4-induced acute liver injury in mice. According to mass spectrometry data, there were 28 chemical compositions in crude and fried Chrysanthemi Flos, mainly including flavonoids and organic acids. 13 compositions such as luteolin, apigenin and luteolin glycoside were increased significantly after frying, while 7 compositions such as chlorogenic acid, luteolin-7-O-glucuronide and apigenin-7-O-glucuronide were decreased significantly after frying. Through principal component analysis, crude and fried Chrysanthemi Flos products were divided into two categories, indicating that there were internal differences in quality. The results of liver injury protection experiment in mice showed that the AST, ALT and MDA contents were significantly decreased and SOD level was increased in mice with liver injury in both the high and medium dose groups. Histopathological examination showed that crude and fried Chrysanthemi Flos can protect the liver by reducing inflammatory cell infiltration, reducing steatosis, and repairing damaged liver cells. The results of this study showed that the chemical compositions had obvious changes after frying, and both crude and fried Chrysanthemis Flos had protective effects on CCl_4-induced acute liver injury in mice. In addition, in the range of high, medium and low doses, the liver protection effect of crude and fried Chrysanthemi Flos increased with the increase of dose. The experiment results provided reference for the mechanism of fried Chrysanthemi Flos and clinical selection of processed products.


Subject(s)
Animals , Mice , Chromatography, High Pressure Liquid , Chrysanthemum , Flavonoids , Flowers , Chemistry , Liver , Chemistry
8.
Chinese Traditional Patent Medicine ; (12): 777-782, 2018.
Article in Chinese | WPRIM | ID: wpr-710236

ABSTRACT

AIM To observe the effect of Zishen Yutai Pills (Cuscutae Semen,Ginseng Radix et Rhizoma,Dipsaci Radix,etc.) on RAS,VEGF,VEGFR-2,sVEGFR-1 and MAPK in recurrent miscarriage mice,and to explore its mechanism.METHODS CBA/J female mice + DBA/2 male mice,and CBA/J female mice + BALB/c male mice were mated by 2 females and 1 male in cage to establish the RSA model and the normal pregnancy CBA × BALB/c mouse model respectively.Since the zeroth day of pregnancy,a total of 24 CBA/J × DBA/2 mice were randomly divided into model control group,Zishen Yutai Pills group and progesterone capsule group,and 10 CBA × BALB/c mice were used as normal pregnancy control group.Mice of all groups after the respective 15-day intervention had their rate of uterine embryo loss measured and calculated.Their pathological changes of decidual tissue were determined by HE staining,their RAS,VEGF,VEGFR-2,sVEGFR-1,MAPK protein and mRNA expressions were detected by immunohistochemistry and real-time PCR.RESULTS Zishen Yutai Pills significantly reduced the rate of embryo loss and improved pathological changes of decidual tissue in RSA mice through regulating mouse decidual tissue angiogenesis and recasting,as revealed by the lowered levels of RAS,VEGF,VEGFR-2 and MAPK,and increased expression of sVEGFR-1.CONCLUSION Zishen Yutai Pills can lower the rate of embryo loss and improve decidual angiogenesis in RSA mice through altering the expression of RAS,VEGF,VEG-FR-2,sVEGFR-1 and MAPK.

9.
Chinese Traditional and Herbal Drugs ; (24): 5264-5270, 2018.
Article in Chinese | WPRIM | ID: wpr-851542

ABSTRACT

Objective To analyze and identify the transitional constituents of combination Psoralea corylifolia-Myristica fragrans in vivo and in vitro, and further study the effect of this combination on transitional components. Methods A rapid ultra-performance liquid chromatography/orthogonal acceleration time-of-flight mass spectrometry (UPLC-Q-TOF/MS) method was established to rapidly analyze constituents of before and after combination P. corylifolia-M. fragrans after oral administration in rats, and combined with Peakview software analysis. Results Compared with in vitro extract of P. corylifolia, there are 15 prototypes absorbed into the blood. Compared with in vitro combination P. corylifolia-M. fragrans extract, 26 prototype components were absorbed into the blood. Compared with M. fragrans extract, six prototype components were absorbed into the blood. Conclusion By using UPLC-Q-TOF/MS method, the main chemical constituents from the combination can be rapidly and accurately identified, and the results would facilitate the quality control of combination P. corylifolia-M. fragrans for safe and efficient use.

10.
China Journal of Chinese Materia Medica ; (24): 1782-1786, 2017.
Article in Chinese | WPRIM | ID: wpr-256096

ABSTRACT

To conduct multiple-reaction monitoring(MRM) quantitative analysis with ultra-high performance liquid chromatography coupled with mass spectrometry method(UPLC-MS/MS), determine the concentrations of psoralen, isopsoralen, bakuchiol and dehydrodiisoeugenol in plasma under positive iron mode with chloramghenicol as internal standard, and investigate the pharmacokinetics process of the main components before and after oral administration of drug pair Psoralea corylifolia -Myristica fragrants. Thirty-six SD rats were randomly divided into three group(A, B, C) and received P. corylifolia extract, P. corylifolia-M. fragrants extract, and M. fragrants extract respectively by intragastric administration. The plasma samples were collected at different time points. In the plasma samples, psoralen, isopsoralen, bakuchiol and dehydrodiisoeugenol showed good linear relationship within concentration rages of 0.098 125 to 39.25, 0.084 37 to 33.75, 0.046 875 to 18.75, and 0.11 to 2.2 mg•L⁻¹ respectively. The precision and stability results showed that the determination method of plasma concentration for such compositions was stable and reliable. The pharmacokinetic parameters obtained by DAS 2.0 showed varying differences before and after compatibility. According to the experimental results, the compatibility of P. corylifolia and M. fragrants can significantly impact the pharmacokinetic process of main components, expand their distribution and accelerate their metabolism and elimination in vivo.

11.
China Journal of Chinese Materia Medica ; (24): 2321-2328, 2016.
Article in Chinese | WPRIM | ID: wpr-236091

ABSTRACT

To observe the effect of total saponins of Clematidis Radix et Rhizoma (TSCR) on serum metabolic profile changes in adjuvant arthritis(AA) rats, and explore its possible action mechanism for AA rats. The AA rat models were induced by Freund's complete adjuvant(FCA), and their histopathological changes were observed. Gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS), principal component analysis(PCA) and partial least squares-discriminant analysis (PLS-DA) were employed to analyze the metabolic profile among normal group, AA model group and TSCR group. Potential biomarkers in the serum were screened based on the variable importance projection(VIP) value>1, P<0.05. As compared with the normal group, 17 potential biomarkers such as aspartic acid, inositol and phenylacetaldehyde were found and identified in the serum of model group rats. As compared with the model group, the above biomarkers were regulated nearly to a normal state after TSCR administration for 16 days. Metabolomic analysis revealed that the total saponins of Clematidis Radix et Rhizoma has a certain therapeutic effect for AA rats, and the mechanism may be related to regulation of lipid metabolism, amino acid metabolism and energy metabolism.

12.
Acta Physiologica Sinica ; (6): 193-200, 2015.
Article in Chinese | WPRIM | ID: wpr-255957

ABSTRACT

Angiotensin II (Ang II) and calcitonin gene-related peptide (CGRP) play important roles in vascular injury and protection. In order to determine the role of CGRP receptor component protein (RCP) in signal transduction whereby CGRP and Ang II mediate the expression of vascular peroxidase-1 (VPO1) in vascular smooth muscle cell (VSMC), mouse derived A10 vascular smooth muscle cell line (A10VSMC) was cultured with CGRP or/and Ang II in vitro. RCP-specific small interference RNA (siRNA-RCP) was used to silence oligonucleotide sequence. Western blot and RT-PCR were used to determine the protein and mRNA expressions of RCP and VPO1, respectively. The results showed that the expressions of RCP and VPO1 were increased in the presence of CGRP or Ang II in the quiescent A10VSMC. But the protein expressions of RCP and VPO1 induced by Ang II were decreased by pretreatment of CGRP (P < 0.05). The expressions of VPO1 were decreased in all the groups treated with siRNA-RCP, compared with those of wide-type counterparts. Meanwhile, the expression of VPO1 was significantly induced by CGRP but not Ang II in the siRNA-RCP treated A10VSMCs. Ang II in combination with CGRP increased the protein expression of VPO1 in the siRNA-RCP-transfected cells, compared with Ang II alone, and this effect could be abolished by catalase. The results suggest that RCP may play an important role in the integration of signal transduction whereby CGRP and Ang II receptors jointly regulate VPO1 expression in VSMC.


Subject(s)
Animals , Mice , Angiotensin II , Pharmacology , Calcitonin Gene-Related Peptide , Pharmacology , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , Metabolism , Peroxidases , Metabolism , RNA, Small Interfering , Signal Transduction
13.
Chinese Journal of Virology ; (6): 668-674, 2014.
Article in Chinese | WPRIM | ID: wpr-280311

ABSTRACT

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.


Subject(s)
Animals , Chick Embryo , Chlorocebus aethiops , Chickens , Coronavirus Infections , Virology , Infectious bronchitis virus , Chemistry , Genetics , Metabolism , Poultry Diseases , Virology , Protein Structure, Tertiary , Spike Glycoprotein, Coronavirus , Chemistry , Genetics , Metabolism , Transfection , Vero Cells
14.
China Journal of Chinese Materia Medica ; (24): 1248-1254, 2014.
Article in Chinese | WPRIM | ID: wpr-321329

ABSTRACT

The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.


Subject(s)
Caffeic Acids , Toxicity , Chemistry, Pharmaceutical , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Toxicity , Quinic Acid , Toxicity , Xanthium , Chemistry , Classification
15.
China Journal of Chinese Materia Medica ; (24): 1766-1771, 2013.
Article in Chinese | WPRIM | ID: wpr-294026

ABSTRACT

This study was establish an UPLC fingerprint of Xanthii Fructus from different habitats, to provide a comprehensive evaluation for its quality control. UPLC-PDA was adopted to analysis of 26 baches of Xanthii Fructus from different habitats. The chromatographic condition was as follow: ACQUITY BEH C18 Column (2.1 mm x 100 mm,1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acid water in gradient mode. The flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 220 nm. The fingerprints of 26 batches Xanthii Fructus were carried out by similarity comparation, cluster and the principal component analysis (PCA). There were nineteen common peaks, nine of which had been identified, and the similarity degrees of the twenty-six batches of the samples were between 0.804 and 0.990. All the samples were classified into six categories, and the PCA value of each fingerprint peak was calculated, and six principal components accounted for over 81. 140% of the total variance were extracted from the original data This method can be used to assess the quality of Xanthii Fructus.


Subject(s)
China , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Ecosystem , Fruit , Chemistry , Quality Control , Xanthium , Chemistry
16.
Chinese Journal of Endemiology ; (6): 182-186, 2012.
Article in Chinese | WPRIM | ID: wpr-643074

ABSTRACT

ObjectiveTo establish the methodology for identification of clinical isolates of Mycobacterium and to identify the distribution of Mycobacterium species in hospitalized patients with tuberculosis in Heilongjiang province.It would provide the basis for accurate diagnosis of infections with Mycobacterium tuberculosis (MTB) and non-tuberculous Mycobacterium (NTM) as well as for effective therapy.Methods Three hundred and thirty Mycobacterium isolates from 330 tuberculosis patients hospitalized and clinically diagnosed in Harbin Chest Hospital from May 2007 to December 2008 were collected.Genomic DNA from the isolates was extracted after inactivation of Mycobacterium.Molecular identification was carried out using PCR,PCR-restriction fragment length polymorphism (RFLP) and sequencing.ResultsAmong 330 clinical isolates,328 were identified as MTB complex (MTBC),accounting for 99.4% (328/330); 2 were NTM,accounting for 0.6% (2/330).Among 328 MTBC isolates,326were MTB,one was Mycobacterium Africanum(M.African) and one was Mycobacterium microti(M,microti),accounting for 99.4% (326/328),0.3% (1/328) and 0.3% (1/328),respectively.It was found that the homology between the two NTM isolates and Mycobacterium avium intracellulare (MAC)was 99% and 93%,respectively,suggesting that the two NTM isolates were MAC.The homology between the two NTM isolates was 93%.ConclusionsPCR plus PCR-RFLP and sequencing provides an ideal method for precise identification of Mycobacterium species.In the clinically diagnosed tuberculosis patients,the predominant Mycobacterium species is MTB,however M.African,M.microti as well as NTM are also found.

17.
Chinese Medical Journal ; (24): 2254-2258, 2010.
Article in English | WPRIM | ID: wpr-237470

ABSTRACT

<p><b>BACKGROUND</b>Therapeutic hypercapnia (TH) has been demonstrated to protect several organs ischemia-reperfusion injury. The study aimed to investigate the effects of therapeutic hypercapnia on hepatic ischemia-reperfusion injury (HIRI).</p><p><b>METHODS</b>Thirty adult male Wistar rats weighing (250+/-20) g were randomized into 3 groups (n=10 in each), group C (control group), group A (hypercapnia group) and group B (CO2 preconditioning group). A segmental ischemia of the liver was induced by interrupting the blood vessels including the bile duct to the median and left lateral lobes for 60 minutes and all the animals were sacrificed after 240 minutes observation period of reperfusion. Mean arterial pressure (MAP) and the blood gases were measured before ischemia (baseline) and at 30, 60, 120, 180 and 240 minutes after reperfusion. Arterial blood samples were obtained for determination of serum levels of TNF-alpha, IL-10, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The histopathology of liver tissues was evaluated by light microscopy. The NF-kappaB expression and apoptotic hepatocytes were respectively determined by immunohistochemistry and TUNEL assay.</p><p><b>RESULTS</b>The serum levels of liver enzymes and TNF-alpha were significantly decreased while the IL-10 level was significantly increased in groups A and B than in group C (P<0.05), and group B surpassed group A (P<0.05). The histopathological scores, the NF-kappaB immunohistochemical score (IHS) and apoptotic index were significantly lower in groups A and B than in group C (P<0.05), and the decrease in group B was more obvious than in group A (P<0.05).</p><p><b>CONCLUSION</b>Therapeutic hypercapnia attenuates ischemia-reperfusion injury to the liver. Moreover, the effects of CO2 preconditioning are outstandingly notable.</p>


Subject(s)
Animals , Male , Rats , Apoptosis , Carbon Dioxide , Therapeutic Uses , Hemodynamics , Immunohistochemistry , In Situ Nick-End Labeling , Inflammation , Drug Therapy , Metabolism , Interleukin-10 , Blood , Liver , Metabolism , Pathology , NF-kappa B , Metabolism , Random Allocation , Rats, Wistar , Reperfusion Injury , Blood , Drug Therapy , Metabolism , Tumor Necrosis Factor-alpha , Blood
18.
Journal of Applied Clinical Pediatrics ; (24)2004.
Article in Chinese | WPRIM | ID: wpr-639651

ABSTRACT

0.05).The scales of subtests including arithmetic,digit span,picture completion,block design and coding of children in CAE group were significantly lower than those of control group(Pa

19.
Chinese Journal of Medical Instrumentation ; (6): 63-64, 2002.
Article in Chinese | WPRIM | ID: wpr-241062

ABSTRACT

A new instrument for measuring pressure pain strength to different bodily parts is introduced in this paper. For a new measuring appliance, it can be used conveniently for detection and evaluation of a patient's pressure pain threshold value, and for the research of bodily channels and network vessels. The system uses silicone oil as pressure conducting medium of a transducer, and five pressure detecting probes with different surface areas are designed to meet the requirements of the parts to be measured. The system has digital lock function for easy operation.


Subject(s)
Humans , Pain , Pain Measurement , Pain Threshold , Physiology , Pressure , Sensitivity and Specificity , Silicone Oils
20.
Chinese Journal of Medical Instrumentation ; (6): 215-217, 2002.
Article in Chinese | WPRIM | ID: wpr-344250

ABSTRACT

This paper is to introduce the method in which the SIEMENS' MR images are converted into BMP format ones based on the correct format analysis of SIMENS' MR images, we condude that the conversion from SIEMENS' MR image to BMP image can be realized by software design.


Subject(s)
Humans , Computers , Magnetic Resonance Imaging , Methods , Software
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