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1.
China Journal of Chinese Materia Medica ; (24): 6423-6433, 2023.
Article in Chinese | WPRIM | ID: wpr-1008842

ABSTRACT

This study aims to investigate the molecular mechanism of tanshinone Ⅱ_(A )(TaⅡ_A) combined with endothelial progenitor cells-derived exosomes(EPCs-exos) in protecting the aortic vascular endothelial cells(AVECs) from oxidative damage via the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt) pathway. The AVECs induced by 1-palmitoyl-2-(5'-oxovaleroyl)-sn-glycero-3-phosphocholine(POVPC) were randomly divided into model, TaⅡ_A, EPCs-exos, and TaⅡ_A+EPCs-exos groups, and the normal cells were taken as the control group. The cell counting kit-8(CCK-8) was used to examine the cell proliferation. The lactate dehydrogenase(LDH) cytotoxicity assay kit, Matrigel assay, DCFH-DA fluorescent probe, and laser confocal microscopy were employed to examine the LDH release, tube-forming ability, cellular reactive oxygen species(ROS) level, and endothelial cell skeleton morphology, respectively. The enzyme-linked immunosorbent assay was employed to measure the expression of interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α. Real-time fluorescence quantitative PCR(qRT-PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of PI3K and Akt. Compared with the control group, the model group showed decreased cell proliferation and tube-forming ability, increased LDH release, elevated ROS level, obvious cytoskeletal disruption, increased expression of IL-1β, IL-6, and TNF-α, and down-regulated mRNA and protein levels of PI3K and Akt. Compared with the model group, TaⅡ_A or EPCs-exos alone increased the cell proliferation and tube-forming ability, reduced LDH release, lowered the ROS level, repaired the damaged skeleton, decreased the expression of IL-1β, IL-6, and TNF-α, and up-regulated the mRNA and protein levels of PI3K and Akt. TaⅡ_A+EPCs-exos outperformed TaⅡ_A or EPCs-exos alone in regulating the above indexes. The results demonstrated that TaⅡ_A and EPCs-exos exerted a protective effect on POVPC-induced AVECs by activating the PI3K/Akt pathway, and the combination of the two had stronger therapeutic effect.


Subject(s)
Proto-Oncogene Proteins c-akt/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Signal Transduction , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/metabolism , Interleukin-6/metabolism , Endothelium, Vascular , Oxidative Stress , Endothelial Progenitor Cells , RNA, Messenger/metabolism , Abietanes
2.
International Eye Science ; (12): 1887-1890, 2023.
Article in Chinese | WPRIM | ID: wpr-996904

ABSTRACT

AIM: To investigate the effect of peripheral defocus spectacles and orthokeratology lenses on the control of axial length in children and adolescents with myopia.METHODS: Prospective study. A total of 71 cases(134 eyes)of children and adolescents with myopia who visited the Second Hospital of Longyan from June 2019 to June 2021 were selected. They were fitted with peripheral defocus spectacles for 12mo and then switched to orthokeratology lenses. The growth of axial length was observed at 3, 6, and 12mo after wearing peripheral defocus spectacles and orthokeratology lenses.RESULTS: The median axial length growth after wearing peripheral defocus spectacles and orthokeratology lenses for 12mo was 0.35 and 0.14mm, respectively. The axial growth at 3, 6, and 12mo after wearing orthokeratology lenses was lower than those after wearing peripheral defocus spectacles(P<0.001), and the growth rate of axial length was significantly reduced. The patients were divided into a rapid progression group(axial growth ≥0.4 mm, 29 cases, 54 eyes)and a non-rapid progression group(axial growth <0.4mm, 42 cases, 80 eyes)according to the axial growth of peripheral defocus spectacles for 12mo. The median axial growth after wearing peripheral defocus spectacles for 12mo in the two groups was 0.70 and 0.24mm, respectively, while the median axial growth after wearing orthokeratology lenses was 0.31 and 0.09mm, respectively. The growth rate was reduced by 56% and 63% respectively in the two groups after wearing orthokeratology lens. The axial growth of cases wearing orthokeratology lenses for 12mo in the non-rapid progression group was lower than that in the rapid progression group, and it did not change with age or diopter. There was no significant difference among different ages and different diopters in the rapid progression group(P>0.05). In the non-rapid progression group, axial growth of cases aged 7-12 years was higher than those aged 13-16 years(P<0.05), but there was no significant difference among different diopters(P>0.05).CONCLUSION: Orthokeratology lens is more effective than peripheral defocus spectacles in controlling axial growth in children and adolescents with myopia, and the control effect of orthokeratology lens on rapid-progressing myopia is remarkable.

3.
China Journal of Chinese Materia Medica ; (24): 4164-4172, 2023.
Article in Chinese | WPRIM | ID: wpr-1008613

ABSTRACT

The study aims to observe the effects and explore the mechanisms of Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination in the treatment of the inflammatory response of mice with atherosclerosis(AS) via the Toll-like receptor 4(TLR4)/myeloid differentiation primary response protein 88(MyD88)/nuclear factor-κB(NF-κB) signaling pathway. Male ApoE~(-/-) mice were randomly assigned into a model group, a Buyang Huanwu Decoction group, an Astragali Radix-Angelicae Sinensis Radix combination group, and an atorvastatin group, and male C57BL/6J mice of the same weeks old were used as the control group. Other groups except the control group were given high-fat diets for 12 weeks to establish the AS model, and drugs were administrated by gavage. Aortic intimal hyperplasia thickness, blood lipid level, plasma inflammatory cytokine levels, M1/M2 macrophage markers, and expression levels of proteins in TLR4/MyD88/NF-κB pathway in the vessel wall were measured to evaluate the effects of drugs on AS lesions and inflammatory responses. The results showed that the AS model was successfully established with the ApoE~(-/-) mice fed with high-fat diets. Compared with the control group, the model group showed elevated plasma total cholesterol(TC), triglyceride(TG), and low-density lipoprotein cholesterol(LDL-c) levels(P<0.05), thickened intima(P<0.01), and increased plasma tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) levels(P<0.01). Moreover, the model group showed increased expression of vascular cell adhesion molecule-1(VCAM-1) and inducible nitric oxide synthase(iNOS)(P<0.01), inhibited expression of endothelial nitric oxide synthase(eNOS) and cluster of differentiation 206(CD206)(P<0.01), and up-regulated mRNA and protein levels of TLR4, MyD88, NF-κB inhibitor alpha(IκBα), and NF-κB in the vessel wall(P<0.05). Compared with the model group, Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination lowered the plasma TC and LDL-c levels(P<0.01), alleviated the intimal hyperplasia(P<0.01), and reduced the plasma TNF-α and IL-6 levels(P<0.05). Moreover, the two interventions promoted the expression of eNOS and CD206(P<0.05), inhibited the expression of VCAM-1 and iNOS(P<0.01), and down-regulated the mRNA and protein levels of TLR4, MyD88, IκBα, and NF-κB(P<0.05) in the vessel wall. This study indicated that Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination could delay the progression of AS, inhibit the polarization of vascular wall macrophages toward M1 type, and attenuate vascular inflammatory response by inhibiting the activation of TLR4/MyD88/NF-κB signaling pathway in the vascular wall. Astragali Radix and Angelicae Sinensis Radix were the main pharmacological substances in Buyang Huanwu Decoction for alleviating the AS vascular inflammatory response.


Subject(s)
Mice , Male , Animals , NF-kappa B/metabolism , Toll-Like Receptor 4/metabolism , NF-KappaB Inhibitor alpha/metabolism , Tumor Necrosis Factor-alpha/metabolism , Interleukin-6/metabolism , Myeloid Differentiation Factor 88/metabolism , Vascular Cell Adhesion Molecule-1/metabolism , Cholesterol, LDL , Hyperplasia , Mice, Inbred C57BL , Atherosclerosis/genetics , Apolipoproteins E/therapeutic use , RNA, Messenger
4.
Chinese Pharmacological Bulletin ; (12): 1182-1189, 2023.
Article in Chinese | WPRIM | ID: wpr-1013794

ABSTRACT

Aim To investigate the antibacterial activity and anti-resistant mutation ability of Qiguiyin decoction (a traditional Chinese herbal formula) combined with levofloxacin against pseudomonas aeruginosa byantibacterial experiment in vitro and serum pharmacology. Methods The minimal inhibitory concentrations (MICs) of levofloxacin and Qiguiyin decoction were detected respectively by the broth dilution technique.The MIC of the combination of two drugs was determined by the micro chessboard dilution method. The effects of combined drugs on enhancing the antibacterial activity of different strains were evaluated respectively by calculating the fractional inhibitory concentration index (FICI). The drug-containing serum of levofloxa-cin group, Qiguiyin decoction group, Qiguiyin decoction combined with levofloxacin group and control group was prepared. The antibacterial rate, MIC and MBC of 10% ~ 90% serum against the two strains were determined. Results Combined with Qiguiyin decoction, MIC of levofloxacin against pseudomonas aeruginosa (standard/resistant) decreased significantly, 0. 5 < FICI

5.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 335-341, 2023.
Article in Chinese | WPRIM | ID: wpr-965850

ABSTRACT

ObjectiveTo discuss the correlation between young patients’ adverse transfusion reaction and pre-transfusion C reactive protein (CRP) level and EO% (percentage of eosinophils) . MethodsThe observation group was chosen from among sixty-six young patients who experienced transfusion-related adverse events between January 2019 and December 2020. For each patient chosen to be included in the observation group, another patient from the same department, with the same disease and gender, who had been hospitalized in the same month and received the same type of blood product transfusion, but had not experienced any transfusion-related adverse effects, was chosen to be in the control group. We examined and compared their ages, transfusion experiences, allergy backgrounds, EO%, and CRP levels in peripheral blood prior to transfusion. A receiver operating characteristic (ROC) curve was used to examine the diagnostic value of EO% and CRP for transfusion-related adverse events. Simultaneously, a logistic analysis was performed on the risk factors for transfusion-related adverse events. ResultsPre-transfusion CRP was higher in patients with FNHTR in the observation group than it was in patients in the control group; pre-transfusion CRP was also higher in patients with ATR in the observation group than it was in patients in the control group. There were also statistically significant differences between these variables in the percentage of patients with transfusion history and pre-transfusion EO% ( P<0.05). For the transfusion of different blood types, there was statistical significance in the occurrence of ATR and FNHTR ( P<0.05). For the diagnosis of FNHTR, the CRP area under the ROC curve was 0.889, and the best cut-off value was 18.05 mg/L. For the diagnosis of ATR, the area under the ROC curve was 0.749, and the best cut-off values were 17.60 mg/L. ConclusionPre-transfusion C-reactive protein level is an independent risk factor for FNHTR and ATR in young patients; the predictive value of EO% for adverse blood transfusion reactions is insufficient.

6.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 73-79, 2021.
Article in Chinese | WPRIM | ID: wpr-905959

ABSTRACT

Objective:To investigate the effect of astragaloside Ⅳ(AST Ⅳ)and Notoginseng total saponins (NTS) combined with bone marrow mesenchymal stem cell (BMSC) transplantation on neural repair and angiogenesis in rats with cerebral ischemia. Method:The rats were randomly divided into a sham operation group, a model group, low- and high-dose AST Ⅳ + NTS groups, a BMSC infusion group, and low- and high-dose BMSC infusion+AST Ⅳ (10 and 20 mg·kg<sup>-1</sup>) + NTS group (25, 50 mg·kg<sup>-1</sup>). BMSCs were isolated and purified by whole bone marrow adherent culture. The positive expression of surface markers of BMSCs (CD29, CD90, CD34, and CD45) was detected by flow cytometry. The focal cerebral ischemia model was established by middle cerebral artery occlusion (MCAO). The PKH26-labeled BMSCs were injected into the tail vein of rats in the BMSC infusion group, once a day. The rats in the combination groups received BMSC injection once a day and intragastric administration of drugs twice a day. Other groups were administered twice a day by gavage. The sham operation group and the model group received the same amount of normal saline. Symptoms and signs of neurological deficits were assessed by the Longa method and the cerebral infarction rate was determined by TTC staining. The survival and vascularization [double positive expression of PKH26/vascular endothelial growth factor (VEGF)] after transplantation of BMSCs were observed by the immunofluorescence method. The protein expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was measured by Western blot. Result:BMSCs were properly isolated and cultured. The identification of surface markers CD29, CD90, CD34, and CD45 was consistent with the characteristics of BMSCs. The neurological deficit score and cerebral infarction rate of the model group were significantly increased (<italic>P</italic><0.01). All drugs and cell transplantation could alleviate the above pathological changes in varying degrees. The strongest effect was observed in high-dose BMSC infusion+AST Ⅳ+NTS group (<italic>P</italic><0.01), which was superior to those in the AST Ⅳ+NTS groups or the BMSC infusion group. BMSC injection helped cells survive in the ischemic brain tissues and promoted angiogenesis, and this effect could be enhanced by the combination with drugs. After cerebral ischemia, the expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was increased, and the effect in the BMSC infusion+AST Ⅳ+NTS groups was the strongest (<italic>P</italic><0.01). Conclusion:AST Ⅳ combined with NTS can promote the survival of transplanted BMSCs and facilitate angiogenesis after target repair of damaged blood vessels after cerebral ischemia. The mechanism may be related to the improvement of the local microenvironment in the brain after cerebral ischemia and the promotion of the survival and differentiation of transplanted stem cells.

7.
International Eye Science ; (12): 2074-2078, 2020.
Article in Chinese | WPRIM | ID: wpr-829708

ABSTRACT

@#The virus-related corneal endotheliitis has diverse inducement, complex etiology, unclear pathogenesis, diverse clinical manifestations. There is no unified classification at present. Auxiliary diagnosis technology is improving constantly, loop-mediated isothermal amplification and all kinds of PCR have a certain auxiliary role in the diagnosis of this disease. But the diagnosis also mainly relys on history and clinical manifestation in clinic. Anti-viral drug and glucocorticoid are the main treatment. Virus-related corneal endotheliitis has high recurrence rate, so there is an urgent need to research new drugs and new treatment. Experiment researches of genetic engineering drugs, such as vaccines, unwinding-primers enzyme inhibitors, and chemokine receptors, bring hope to the treatment of virus-related corneal endotheliitis. The purpose of this article is to review the related research of virus-related corneal endotheliitis.

8.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 44-55, 2020.
Article in Chinese | WPRIM | ID: wpr-872697

ABSTRACT

The prescription research and clinical application of Wenjingtang were summarized in order to provide reference for the formulation of material standard and the development of compound preparation. By systematically sorting out the relevant ancient medical books and modern literature reports, combined with the relevant policy requirements of the development of compound preparations of famous classical formulas, this paper expounded the existing problems and put forward some suggestions. Wenjingtang is composed of nine herbs, which is derived from Complete Effective Prescriptions for Women's Diseases written by CHEN Zi-ming in Song dynasty. The original medicinal plants and medicinal parts of the formula were basically the same as those recorded in the 2020 edition of Chinese Pharmacopoeia, for example, Paeonia lactiflora was the choice of Paeoniae Radix Alba, Cinnamomi Cortex was selected as Guixin, the rhizoma of Curcuma phaeocaulis was selected as Curcumae Rhizoma. It was suggested that raw products should be selected for decoction pieces and processed according to the methods recorded in the 2020 edition of Chinese Pharmacopoeia. According to the dosage of 1 Liang=40 g, the dosages of Angelicae Sinensis Radix, Chuanxiong Rhizoma, Paeoniae Radix Alba, Cinnamomi Cortex, Moutan Cortex and Curcumae Rhizoma were 20 g, the dosages of Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma and Achyranthis Bidentatae Radix were 40 g, the total amount of this formula was 240 g. The 9 kinds of decoction pieces were crushed (particle size<6 mm) separately and mixed, each dose was 20 g, 525 mL of water was added, and then decocted to 280 mL. After filtration, warm medicine was taken for once a day. According to ancient books, Wenjingtang has the functions of activating blood circulation, regulating menstruation,warming meridians and dispersing cold. It is mainly used to treat dysmenorrhea, irregular menstruation and other diseases in modern clinical practice. In addition, it has certain curative effect on endometriosis, pelvic inflammatory disease, acne, eczema and other skin diseases. Through the research, the historical evolution and clinical application of Wenjingtang are fully clarified, which can provide research for the later development and application of this famous classical formula.

9.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 229-234, 2019.
Article in Chinese | WPRIM | ID: wpr-801891

ABSTRACT

Mori Cortex is the dry root bark of Moras alba L. and usually used in clinical practice. It is sweet and cold in nature, and enters the lung meridian. With effects in purging lung and relieving asthma, and inducing diuresis to reduce edema, it is mainly used to treat lung heat, asthma, cough, swelling, urine deficiency and facial skin edema. In clinic, it is mainly used for the treatment of respiratory system, urinary infection and diabetes mellitus. In recent years, great progress has been made in studies on the pharmacological effects of Mori Cortex. The literatures on the pharmacological effects of Mori Cortex in recent years were reviewed and summarized in this paper. Mori Cortex has antitussive, anti-inflammatory, hypoglycemic, cardiovascular, antiviral, anticancer, immunoregulatory, antioxidation and anti-allergy and other pharmacological effects, in addition to antitussive, expectorant, antiasthmatic and other traditional effects. Total flavones have a strong pharmacological activity. These extended studies provide valuable reference for the further development of Mori Cortex. This paper summarizes the pharmacological effects of Mori Cortex, proposes the key directions of further studies, and provides the beneficial reference for better development and utilization of Mori Cortex.

10.
Journal of Acupuncture and Tuina Science ; (6): 16-23, 2019.
Article in Chinese | WPRIM | ID: wpr-746361

ABSTRACT

Objective:To observe the therapeutic effect of electroacupuncture (EA) at Zusanli (ST 36),Guanyuan (CV 4) and Ashi points on adjuvant arthritis rats,and explore the mechanism of EA treatment of rheumatoid arthritis (RA).Methods:Sixty male rats were randomly divided into a normal group,a model group,a methotrexate group and an EA group,with 15 rats in each group.Rats in the normal group and the model group were routinely raised and did not receive treatment;rats in the methotrexate group received methotrexate at a dose of 0.35 mg/(kg·bw),twice a week for 3 weeks;rats in the EA group received acupuncture at Zusanli (ST 36),Guanyuan (CV 4) and Ashi points,and the bilateral Zusanli (ST 36) and Ashi points were connected to EA apparatus,once a day for 3 weeks.The general status,the swelling degree of the toe,the arthritis index (AI) score,the pathological morphology of the ankle joint,and the mRNA expressions of cellular inhibitor of apoptosis protein (c-IAP) 1 and c-IAP2 in joint synovial tissue cells of the rats in each group were observed.Results:The swelling degree of the toe,AI score and mRNA expressions of c-IAP1 and c-IAP2 in the model group were significantly higher than those in the normal group (all P<0.05).Compared with the model group,the swelling degree of the toe,AI score and mRNA expressions of c-IAP1 and c-IAP2 in the methotrexate group and the EA group improved (P<0.01 or P<0.05);the expressions of c-IAP1 mRNA and c-IAP2 mRNA in rat synovial tissues in the EA group were significantly higher than those in the methotrexate group (P<0.01).Conclusion:EA alleviates joint swelling in rats with adjuvant arthritis.The mechanism may be related to suppressing mRNA expressions of c-IAP1 and c-IAP2,thus to induce apoptosis of synoviocytes.

11.
Chinese Medical Journal ; (24): 2683-2690, 2016.
Article in English | WPRIM | ID: wpr-230900

ABSTRACT

<p><b>BACKGROUND</b>Among HIV-infected patients initiating antiretroviral therapy (ART), early changes in CD4+ T-cell subsets are well described. However, HIV-infected late presenters initiating treatment present with a suboptimal CD4+ T-cell reconstitution and remain at a higher risk for AIDS and non-AIDS events. Therefore, factors associated with CD4+ T-cell reconstitution need to be determined in this population, which will allow designing effective immunotherapeutic strategies.</p><p><b>METHODS</b>Thirty-one adult patients with baseline CD4+ T-cell count <350 cells/mm3 exhibiting viral suppression after ART initiation were followed in the HIV/AIDS research center of Peking Union Medical College Hospital in Beijing, China, from October 2002 to September 2013. Changes in T-cell subsets and associated determinants were measured.</p><p><b>RESULTS</b>Median baseline CD4+ T-cell count was 70 cells/mm3. We found a biphasic reconstitution of T-cell subsets and immune activation: a rapid change during the first 6 months followed by a more gradual change over the subsequent 8 years. Baseline CD4+ T-cell count >200 cells/mm3 in comparison to CD4+ T-cell count ≤200 cells/mm3 was associated with more complete immune Reconstitution (77.8% vs. 27.3% respectively; P = 0.017) and normalized CD4/CD8 ratio. We showed that the baseline percentage of naive CD4+ T-cell was a predictive marker for complete immune reconstitution (area under receiver operating characteristic curve 0.907), and 12.4% as cutoff value had a sensitivity of 84.6% and a specificity of 88.2%.</p><p><b>CONCLUSIONS</b>Baseline naive CD4+ T-cell percentage may serve as a predictive marker for optimal immune reconstitution during long-term therapy. Such study findings suggest that increasing thymic output should represent an avenue to improve patients who are diagnosed late in the course of infection.</p>


Subject(s)
Adult , Female , Humans , Male , Antiretroviral Therapy, Highly Active , Methods , CD4 Lymphocyte Count , CD4-CD8 Ratio , CD4-Positive T-Lymphocytes , Metabolism , HIV Infections , Drug Therapy , Allergy and Immunology , Metabolism , HIV-1 , Allergy and Immunology , Virulence , Prospective Studies , T-Lymphocyte Subsets , Allergy and Immunology
12.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 910-915, 2016.
Article in English | WPRIM | ID: wpr-238423

ABSTRACT

This study investigated the molecular markers of DS-1-47, a component of an implantation- promoting traditional Chinese medicine consisting of Astragalus mongholicus, Atractylodes macrocephala, Scutellaria baicalensis and Dipsacales, in an attempt to clarify the molecular mechanism and action targets of DS-1-47. Controlled ovarian stimulation (COS) method was used to establish the implantation dysfunction models of mice. Animals were divided into normal pregnant group, COS model group and DS-1-47 group. Laser capture microdissection-double dimensional electrophoresis-mass spectrum (LCM-DE-MS) was used to analyze the uterine protein molecules that were possibly involved in the promotion of implantation. Twenty-three proteins in DS-1-47 group were significantly changed as compared to those in COS model group, with 7 proteins down-regulated and 16 proteins up-regulated. Except for some constituent proteins, the down-regulated proteins included collagen α-1 (VI) chain, keratin 7, keratin 14, myosin regulatory light chain 12B, myosin light polypeptide 9, heat shock protein β-7, and C-U-editing enzyme APOBEC-2; the up-regulated proteins included apolipoprotein A-I, calcium regulated protein-3, proliferating cell nuclear antigen, L-xylulose reductase, and calcium binding protein. These 23 proteins that were regulated by DS-1-47 represented a broad diversity of molecule functions. The down-regulated proteins were associated with stress and immune response, and those up-regulated proteins were related to proliferation. It was suggested that these proteins were important in regulating the uterine environment for the blastocyst implantation. By identification of DS-1-47 markers, proteomic analysis coupled with functional assays is demonstrated to be a promising approach to better understand the molecular mechanism of traditional Chinese medicine.


Subject(s)
Animals , Female , Mice , Pregnancy , Drugs, Chinese Herbal , Pharmacology , Embryo Implantation , Ovulation Induction , Proteome , Genetics , Metabolism , Uterus , Metabolism , Physiology
13.
China Journal of Chinese Materia Medica ; (24): 1227-1234, 2016.
Article in Chinese | WPRIM | ID: wpr-320873

ABSTRACT

High throughput sequencing technology is also called Next Generation Sequencing (NGS), which can sequence hundreds and thousands sequences in different samples at the same time. In the present study, the culture-independent high throughput sequencing technology was applied to sequence the fungi metagenomic DNA of the fungal internal transcribed spacer 1(ITS 1) in the root of Sinopodophyllum hexandrum. Sequencing data suggested that after the quality control, 22 565 reads were remained. Cluster similarity analysis was done based on 97% sequence similarity, which obtained 517 OTUs for the three samples (LD1, LD2 and LD3). All the fungi which identified from all the reads of OTUs based on 0.8 classification thresholds using the software of RDP classifier were classified as 13 classes, 35 orders, 44 family, 55 genera. Among these genera, the genus of Tetracladium was the dominant genera in all samples(35.49%, 68.55% and 12.96%).The Shannon's diversity indices and the Simpson indices of the endophytic fungi in the samples ranged from 1.75-2.92, 0.11-0.32, respectively.This is the first time for applying high through put sequencing technol-ogyto analyze the community composition and diversity of endophytic fungi in the medicinal plant, and the results showed that there were hyper diver sity and high community composition complexity of endophytic fungi in the root of S. hexandrum. It is also proved that the high through put sequencing technology has great advantage for analyzing ecommunity composition and diversity of endophtye in the plant.

14.
China Journal of Chinese Materia Medica ; (24): 1837-1841, 2015.
Article in Chinese | WPRIM | ID: wpr-351254

ABSTRACT

The contents of two lignans, namely 4'-demethylpodophyllotoxin and podophyllotoxin in cultivated and wild Sinopodophyllum hexandrum plants were extracted by ultrasonicaction and determined by HPLC. According to the result showed, the order of parts of cultivated plants containing 4'-demethylpodophyllotoxin from high to low is as follows: stem > root, no 4'-demethypodophyllotoxin was detected in leaves of cultivated plants; The order of parts of wild plants 4'-demethylpodophyllotoxin from high to low is as follows: lateral root > petiole > rhizome > leaf, no 4'-demethypodophyllotoxin was detected in fruit. The order of parts of cultivated and wild S. hexandrum containing podophyllotoxin from high to low is as follows: root > petiole > leaf ( > fruit). Both of the lignan contents in different parts of cultivated plant varied in a " W" curve with the changes in seasons, with the highest content in July.


Subject(s)
Berberidaceae , Chemistry , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fruit , Chemistry , Lignans , Plant Leaves , Chemistry , Plant Roots , Chemistry , Plant Stems , Chemistry , Rhizome , Chemistry , Seasons
15.
Chinese Journal of Contemporary Pediatrics ; (12): 190-195, 2015.
Article in Chinese | WPRIM | ID: wpr-346185

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of 1,25-(OH)(2)D(3) on airway remodeling and expression of high mobility group box 1 (HMGB1) and IL-17 in asthmatic mice.</p><p><b>METHODS</b>Fifty female mice were randomly divided into 5 groups: control, asthma, low-dose, middle-dose, and high-dose intervention groups (n=10 each). Asthma was induced by intraperitoneal injections of ovalbumin (OVA) and aerosol inhalation of OVA solution. The low-dose, middle-dose, and high-dose intervention groups were administered with 1,25-(OH)(2)D(3) solution at the dosage of 1, 4 and 10 μg/kg respectively by intraperitoneal injections before asthma challenge. The airway structural changes were assessed by hematoxylin and eosin staining. mRNA expression levels of HMGB1 and IL-17 in the lung tissues were evaluated by RT-PCR. The protein levels of HMGB1 and IL-17 in the lung tissues were observed by immunohistochemistry.</p><p><b>RESULTS</b>The airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 were higher in the untreated asthma group than in the control group (P<0.05). The airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 were lower in the middle-dose and low-dose intervention groups than in the untreated asthma group, and the middle-dose intervention group demonstrated lower airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 than in the low-dose intervention group (P<0.05). However, the airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 in the high-dose intervention group were higher than in the untreated asthma group (P<0.05).</p><p><b>CONCLUSIONS</b>HMGB1 and IL-17 may be involved in the airway remodeling process in asthmatic mice. A moderate amount of HMGB1 and IL-17 may be involved in the airway remodeling process in asthmatic mice. A moderate amount of 1,25-(OH)(2)D(3) can improve the airway remodeling, but a higher dose of 1,25-(OH)(2)D(3) may affect adversely the airway remodeling process.</p>


Subject(s)
Animals , Female , Mice , Airway Remodeling , Asthma , Drug Therapy , Metabolism , Pathology , Calcitriol , Pharmacology , Dose-Response Relationship, Drug , HMGB1 Protein , Genetics , Physiology , Interleukin-17 , Genetics , Physiology , Lung , Metabolism , Pathology , Mice, Inbred BALB C
16.
China Journal of Chinese Materia Medica ; (24): 1209-1213, 2014.
Article in Chinese | WPRIM | ID: wpr-321337

ABSTRACT

<p><b>OBJECTIVE</b>To construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.</p><p><b>METHOD</b>The 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.</p><p><b>RESULT</b>Plant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.</p><p><b>CONCLUSION</b>Genetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.</p>


Subject(s)
Anti-Bacterial Agents , Pharmacology , Biomarkers , Cinnamates , Pharmacology , Escherichia coli , Genetics , Escherichia coli Proteins , Genetics , Metabolism , Gene Expression , Genetic Vectors , Genetics , Metabolism , Hygromycin B , Pharmacology , Mannose-6-Phosphate Isomerase , Genetics , Metabolism , Plants, Genetically Modified , Genetics , Metabolism , Salvia miltiorrhiza , Genetics , Metabolism , Transformation, Genetic
17.
Chinese Journal of Epidemiology ; (12): 998-1002, 2013.
Article in Chinese | WPRIM | ID: wpr-320954

ABSTRACT

<p><b>OBJECTIVE</b>To obtain information on HIV disease progression through western blot (WB) band profile.</p><p><b>METHODS</b>1466 available serum samples of newly reported HIV/AIDS cases, confirmed by WB from Dehong prefecture of Yunnan province in 2011, were tested, using the BED HIV Incidence Capture Enzyme Immunoassay (BED-CEIA) and Limiting Antigen Avidity Enzyme Immunoassay (LAg-Avidity EIA), respectively. Differences of WB band profiled between recent infections and established infections were then compared.</p><p><b>RESULTS</b>Infections defined as 'recent' showed lower mean antibody intensities to p17, p24, p31, gp41, p51, p55, p66, gp120 and gp160 in WB when compared to those established infections with statistical difference(P < 0.05).</p><p><b>CONCLUSION</b>These WB band profiles which were considered as 'recent' infections would include factors as:lack of antibody reactivity to gp41 antigen, with intensity scores of all bands lower than 13, and those intensity scores of antibody to env protein lower than 5 or to pol protein lower than 4.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Acquired Immunodeficiency Syndrome , Blood , Diagnosis , Epidemiology , Blotting, Western , China , HIV Seropositivity , HIV-1 , Allergy and Immunology
18.
Chinese Journal of Applied Physiology ; (6): 116-127, 2013.
Article in Chinese | WPRIM | ID: wpr-358664

ABSTRACT

<p><b>OBJECTIVE</b>To observe the correlation between the decline of cognitive function and the level of plasma homocysteine in patients with Alzheimer's disease (AD).</p><p><b>METHODS</b>Thirty six AD patients were selected from hospitals in Tianjin. The enrolled patients were in accord with the diagnosis criteria. Thirty two control subjects were corresponding patients without AD in the period. Blood samples were extracted from each subject to determine the levels of homocysteine (Hcy) and folate. Cognitive status was evaluated by the mini- mental state examination (MMSE) and clinical dementia rating scale (CDR).</p><p><b>RESULTS</b>The mean value of serum Hcy concentration [(17.51 +/- 5.62) micromol/L] of AD group was higher than that of control group [(12.38 +/- 4.25)micromol/L]. The serum [(5.17 +/- 1.76) microg/L] and diet folate [(206.94 +/- 44.51) microg/d] concentration of AD group were lower than those of control group [(7.92 +/- 2.22) microg/L, (259.74 +/- 41.92) microg/ d]. The incidence of hyperhomocysteinemia in AD group (64%) was higher than that in control group (22%). A significant relation between Hcy concentrations and the CDR was observed. With the increase of Hcy concentrations the CDR raised, and with the increase of Hcy concentrations the MMSE decreased.</p><p><b>CONCLUSION</b>Hyperhomocysteinemia is one of the risk factors inducing the onset of AD. There is a significant negative correlation between Hcy levels and cognitive levels in AD group. Folate deficiency is an important reason to cause elevated Hcy levels in AD.</p>


Subject(s)
Humans , Alzheimer Disease , Blood , Case-Control Studies , Folic Acid , Blood , Homocysteine , Blood , Hyperhomocysteinemia , Blood
19.
China Journal of Chinese Materia Medica ; (24): 817-820, 2013.
Article in Chinese | WPRIM | ID: wpr-318638

ABSTRACT

<p><b>OBJECTIVE</b>To prepare the new traditional Chinese medicine preparation--pH-dependent brevisapin colon-specific tablets, and investigate its in vitro release, in order to discuss the feasibility of preparing colon-targeted traditional Chinese medicines.</p><p><b>METHOD</b>With scutellarin, the active ingredient in brevisapin, as the evaluation index, coating prescriptions of the preparation was screened. The in vitro release determination method was adopted to detect the in vitro release performance of the preparation.</p><p><b>RESULT</b>The in vitro release determination results showed no brevisapin in artificial pH 1. 2 dilute hydrochloric acid solution for 2 h, an accumulated dissolution rate of less than 5% in pH 6. 8 phosphate buffer solution for 4 h, but an accumulated dissolution rate exceeding 90% in pH 7. 6 phosphate buffer solution for 1 h.</p><p><b>CONCLUSION</b>Brevisapin colon-specific tablets prepared can realize colon-specific release.</p>


Subject(s)
Administration, Oral , Apigenin , Chemistry , Buffers , Chemistry, Pharmaceutical , Methods , Colon , Drugs, Chinese Herbal , Chemistry , Glucuronates , Chemistry , Hydrochloric Acid , Chemistry , Hydrogen-Ion Concentration , Organ Specificity , Solubility , Tablets
20.
China Journal of Chinese Materia Medica ; (24): 678-682, 2013.
Article in Chinese | WPRIM | ID: wpr-350707

ABSTRACT

<p><b>OBJECTIVE</b>To study the application of degrading multi-enzymes from Ganoderma lucidum in extracting effective constituents from fibrous roots of Salvia miltiorrhiza.</p><p><b>METHOD</b>Effective constituents were extracted from fibrous roots by degrading multi-enzymes of wood fiber. The enzymatic parameters were optimized by the orthogonal design.</p><p><b>RESULT</b>The extraction efficiencies of total tanshinones and total salvianolic acids in the extracts of fibrous roots of S. miltiorrhiza was obtained using optimum enzymolysis process reached 11.923%, 12.465%, respectively, which were 62.794%, 56.086% more than that by conventional non-enzymatic hydrolysis.</p><p><b>CONCLUSION</b>Degrading multi-enzymes of wood fiber can be used to fully extract effective constituents from fibrous roots of S. miltiorrhiza, which provides a new approach for recycling wastes of traditional Chinese medicines.</p>


Subject(s)
Alkenes , Metabolism , Abietanes , Metabolism , Drugs, Chinese Herbal , Metabolism , Hydrogen-Ion Concentration , Plant Roots , Chemistry , Polyphenols , Metabolism , Reishi , Salvia miltiorrhiza , Chemistry , Temperature , Wood
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