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1.
Chinese Journal of Neurology ; (12): 1413-1418, 2022.
Article in Chinese | WPRIM | ID: wpr-958046

ABSTRACT

Systemic lupus erythematosus (SLE) complicated with Aspergillus fumigatus brain abscess is rare and needs to be differentiated from bacterial brain abscess and neuropsychiatric lupus. This article reports 2 cases of surgically diagnosed SLE combined with Aspergillus fumigatus brain abscess. The first patient was a 34-year-old woman. Six months after the diagnosis of SLE, she developed convulsions and unconsciousness. She was diagnosed as neuropsychiatric lupus at the first hospitalization because of negative imaging. After discharge, repeated head magnetic resonance imaging revealed abscess-like signals. The second patient, a 20-year-old male, developed high fever, convulsions, and unconsciousness 3 years after the diagnosis of SLE, and head imaging showed an abscess-like signal. The etiology of the cerebrospinal fluid of the 2 patients was both negative, and the Aspergillus fumigatus brain abscess was diagnosed by pathology through abscess resection or drainage. After treatment with voriconazole, the symptoms were relieved and the lesions were subsided.

2.
Protein & Cell ; (12): 641-660, 2020.
Article in English | WPRIM | ID: wpr-828754

ABSTRACT

In mammalian cells, long noncoding RNAs (lncRNAs) form complexes with proteins to execute various biological functions such as gene transcription, RNA processing and other signaling activities. However, methods to track endogenous lncRNA dynamics in live cells and screen for lncRNA interacting proteins are limited. Here, we report the development of CERTIS (CRISPR-mediated Endogenous lncRNA Tracking and Immunoprecipitation System) to visualize and isolate endogenous lncRNA, by precisely inserting a 24-repeat MS2 tag into the distal end of lncRNA locus through the CRISPR/Cas9 technology. In this study, we show that CERTIS effectively labeled the paraspeckle lncRNA NEAT1 without disturbing its physiological properties and could monitor the endogenous expression variation of NEAT1. In addition, CERTIS displayed superior performance on both short- and long-term tracking of NEAT1 dynamics in live cells. We found that NEAT1 and paraspeckles were sensitive to topoisomerase I specific inhibitors. Moreover, RNA Immunoprecipitation (RIP) of the MS2-tagged NEAT1 lncRNA successfully revealed several new protein components of paraspeckle. Our results support CERTIS as a tool suitable to track both spatial and temporal lncRNA regulation in live cells as well as study the lncRNA-protein interactomes.

3.
Protein & Cell ; (12): 641-660, 2020.
Article in English | WPRIM | ID: wpr-828590

ABSTRACT

In mammalian cells, long noncoding RNAs (lncRNAs) form complexes with proteins to execute various biological functions such as gene transcription, RNA processing and other signaling activities. However, methods to track endogenous lncRNA dynamics in live cells and screen for lncRNA interacting proteins are limited. Here, we report the development of CERTIS (CRISPR-mediated Endogenous lncRNA Tracking and Immunoprecipitation System) to visualize and isolate endogenous lncRNA, by precisely inserting a 24-repeat MS2 tag into the distal end of lncRNA locus through the CRISPR/Cas9 technology. In this study, we show that CERTIS effectively labeled the paraspeckle lncRNA NEAT1 without disturbing its physiological properties and could monitor the endogenous expression variation of NEAT1. In addition, CERTIS displayed superior performance on both short- and long-term tracking of NEAT1 dynamics in live cells. We found that NEAT1 and paraspeckles were sensitive to topoisomerase I specific inhibitors. Moreover, RNA Immunoprecipitation (RIP) of the MS2-tagged NEAT1 lncRNA successfully revealed several new protein components of paraspeckle. Our results support CERTIS as a tool suitable to track both spatial and temporal lncRNA regulation in live cells as well as study the lncRNA-protein interactomes.

4.
Protein & Cell ; (12): 641-660, 2020.
Article in English | WPRIM | ID: wpr-827024

ABSTRACT

In mammalian cells, long noncoding RNAs (lncRNAs) form complexes with proteins to execute various biological functions such as gene transcription, RNA processing and other signaling activities. However, methods to track endogenous lncRNA dynamics in live cells and screen for lncRNA interacting proteins are limited. Here, we report the development of CERTIS (CRISPR-mediated Endogenous lncRNA Tracking and Immunoprecipitation System) to visualize and isolate endogenous lncRNA, by precisely inserting a 24-repeat MS2 tag into the distal end of lncRNA locus through the CRISPR/Cas9 technology. In this study, we show that CERTIS effectively labeled the paraspeckle lncRNA NEAT1 without disturbing its physiological properties and could monitor the endogenous expression variation of NEAT1. In addition, CERTIS displayed superior performance on both short- and long-term tracking of NEAT1 dynamics in live cells. We found that NEAT1 and paraspeckles were sensitive to topoisomerase I specific inhibitors. Moreover, RNA Immunoprecipitation (RIP) of the MS2-tagged NEAT1 lncRNA successfully revealed several new protein components of paraspeckle. Our results support CERTIS as a tool suitable to track both spatial and temporal lncRNA regulation in live cells as well as study the lncRNA-protein interactomes.

5.
Chinese Acupuncture & Moxibustion ; (12): 1189-1194, 2018.
Article in Chinese | WPRIM | ID: wpr-777305

ABSTRACT

OBJECTIVE@#To study the effects of moxibustion at different time points on serum level of tumor necrosis factor-α (TNF-α) in rats with rheumatoid arthritis (RA), and to explore its regulation mechanism on circadian rhythm.@*METHODS@#A total of 96 Sprague-Dawley (SD) adult rats were randomly assigned into a blank group, a model group, a moxibustion at 5-7 AM group and a moxibustion at 5-7 PM group, 24 rats in each group, half male and half female. Each group was further divided into a 0 AM group, a 6 AM group, a 12 N group and a 6 PM group, 6 rats in each group. All rats were treated with the 12 h/12 h light-dark cycle in the whole process of experiment. Except for the blank group, all rats were treated with intracutaneous injection of freund's complete adjuvant (FCA) at right foot pad to establish the RA model. The rats at the two moxibustion groups were treated with grain-sized moxibustion at "Shenshu" (BL 23) and "Zusanli" (ST 36) at 5-7 AM and 5-7 PM, respectively, one side per treatment, once a day; six treatments were taken as one course and 3 courses were given with an interval of one day between courses. The rats in the remaining groups were treated with identical fixation but without moxibustion intervention. The right foot volume was measured before model establishment, after model establishment and after treatment. The blood samples were collected after treatment and the serum level of TNF-α was measured by ELISA. The SPSS 21.0 software and Halberg Cosinor were adopted to analyze the experiment data.@*RESULTS@#After treatment, compared with the blank group, the foot swelling severity was significantly increased in the model group, moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (all <0.01); compared with the model group, the foot swelling severity was significantly reduced in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (both <0.01). Compared with the blank group, the serum level of TNF-α was increased significantly in the model group and moxibustion at 5-7 AM group (both <0.05); compared with the model group, the serum level of TNF-α was reduced significantly in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (both <0.05). The serum level of TNF-α showed circadian rhythm in all the groups (all <0.05), and the peak appeared at night phase; compared with the blank group, the median value of TNF-α was increased significantly in the model group (<0.05), the peak phase was delayed and the amplitude was increased (<0.05); compared with the model group, the median value of TNF-α was significantly reduced in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (<0.01), the peak phase was advanced and the amplitude was reduced (<0.05).@*CONCLUSION@#Moxibustion could effectively reduce the serum level of TNF-α to relieve the foot swelling severity in RA rats. Moxibustion could regulate the circadian rhythm of TNF-α to play its effects on the inhibition of the synthesis of TNF-α. No efficacy is observed between the treatment at 5-7 AM and 5-7 PM.


Subject(s)
Animals , Female , Male , Rats , Acupuncture Points , Arthritis, Rheumatoid , Circadian Rhythm , Moxibustion , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha
6.
Journal of Central South University(Medical Sciences) ; (12): 410-414, 2018.
Article in Chinese | WPRIM | ID: wpr-693831

ABSTRACT

Objective:To evaluate the incidence and microbiological profile of meningitis and/or bacteremia after craniotomy in patients with glioma and analyze the risk factors relevant to postoperative meningitis.Methods:All demographic data,etiological data and clinical data for hospitalized patients who underwent craniotomy in Dept.of Neurosurgery of Peking Union Medical College Hospital between Jan.2013 to Dec.2017 were recorded.Results:The incidence of post-craniotomy meningitiswas 7.73%,bacteremia was 2.21% and meningitis combined with bacteremia was 1.10%.The positive rate for cerebrospinal fluid culture was 78.57%.Among the 11 cases of meningitis,20 bacteria were detected,and the most common gram-positive bacteria were:epidermal Staphylococcus (3 case,15.0%),and hemolytic Staphylococcus (3 case,15.0%);the most common gram-negative one was Klebsiella pneumoniae (2 case,10.0%).Among the 5 cases of bacteremia,Klebsiella pneumoniae was the most common one (3 cases,60.0%).Most of the gram-positive bacteria were sensitive to vancomycin and were resistant to penicillin G.Most of the gram-negative bacteria were sensitive to the 3rd generation of cephalosporin and meropenem.Among all the 16 patients,13 were good in prognosis (81.3%) while 3 (18.8%) were poor in prognosis,with 2 deaths and 1 discharged automatically.Among the 11 patients with meningitis,7 patients (63.6%) were male and 7(63.6%) were older than 50 years old.The lesionsin the 6 patients located in the frontal lobe (54.5%) and the glioma in 6 patients was at WHO grade Ⅳ (54.5%).Three patients were diagnosed as diabetes before the surgery (27.3%).Conclusion:Postoperative meningitis and bacteremia can occur in patients with glioma.The pathogens and drug susceptibility are different.The risk factors for meningitis after craniotomy should be concerned before making a treatment plan.

7.
Genomics, Proteomics & Bioinformatics ; (4): 294-306, 2018.
Article in English | WPRIM | ID: wpr-772979

ABSTRACT

Protein nitration and nitrosylation are essential post-translational modifications (PTMs) involved in many fundamental cellular processes. Recent studies have revealed that excessive levels of nitration and nitrosylation in some critical proteins are linked to numerous chronic diseases. Therefore, the identification of substrates that undergo such modifications in a site-specific manner is an important research topic in the community and will provide candidates for targeted therapy. In this study, we aimed to develop a computational tool for predicting nitration and nitrosylation sites in proteins. We first constructed four types of encoding features, including positional amino acid distributions, sequence contextual dependencies, physicochemical properties, and position-specific scoring features, to represent the modified residues. Based on these encoding features, we established a predictor called DeepNitro using deep learning methods for predicting protein nitration and nitrosylation. Using n-fold cross-validation, our evaluation shows great AUC values for DeepNitro, 0.65 for tyrosine nitration, 0.80 for tryptophan nitration, and 0.70 for cysteine nitrosylation, respectively, demonstrating the robustness and reliability of our tool. Also, when tested in the independent dataset, DeepNitro is substantially superior to other similar tools with a 7%-42% improvement in the prediction performance. Taken together, the application of deep learning method and novel encoding schemes, especially the position-specific scoring feature, greatly improves the accuracy of nitration and nitrosylation site prediction and may facilitate the prediction of other PTM sites. DeepNitro is implemented in JAVA and PHP and is freely available for academic research at http://deepnitro.renlab.org.


Subject(s)
Humans , Amino Acid Sequence , Amino Acids , Metabolism , Deep Learning , Internet , Neural Networks, Computer , Nitrosation , Proteins , Chemistry , Metabolism , Reproducibility of Results , Software
8.
The Journal of Practical Medicine ; (24): 1021-1024, 2017.
Article in Chinese | WPRIM | ID: wpr-619005

ABSTRACT

Objective To investigate the effects and the possible mechanisms of nerve growth factor (NGF) on serum levels of vascular endothelial growth factor (VEGF) and stromal cell-derived factor 1 (SDF-1) protein expression in rats with focal cerebral ischemia.Methods Male Sprague-Dawley rats weighting 200 ~ 250 g were subjected to middle cerebral occlusion (MCAO).MCAO rats were randomly divided into NGF group,saline control group and NGF+PI3K antagonist group (n =12).The neurological function was evaluated on the 4th,7th day after MCAO,and the serum levels of VEGF and SDF-1 protein were measured by ELISA.Results The neurological function was better in rats of the NGF group than those of the saline control group and NGF+PI3K antagonist on the 4th,7th day after MCAO (P < 0.05).The serum levels of VEGF and SDF-1 protein of the NGF group was significantly higher than that of the saline control group and NGF+PI3K antagonist group on the 4th day after MCAO (P < 0.05).Conclusions Treatment with NGF may improve neurological function of rats with focal cerebral ischemia,and upregulate serum levels of VEGF and SDF-1 protein expression.PI3K/AKT signal pathway may have attended the above regulation.

9.
Protein & Cell ; (12): 601-611, 2017.
Article in English | WPRIM | ID: wpr-756965

ABSTRACT

Targeted point mutagenesis through homologous recombination has been widely used in genetic studies and holds considerable promise for repairing disease-causing mutations in patients. However, problems such as mosaicism and low mutagenesis efficiency continue to pose challenges to clinical application of such approaches. Recently, a base editor (BE) system built on cytidine (C) deaminase and CRISPR/Cas9 technology was developed as an alternative method for targeted point mutagenesis in plant, yeast, and human cells. Base editors convert C in the deamination window to thymidine (T) efficiently, however, it remains unclear whether targeted base editing in mouse embryos is feasible. In this report, we generated a modified high-fidelity version of base editor 2 (HF2-BE2), and investigated its base editing efficacy in mouse embryos. We found that HF2-BE2 could convert C to T efficiently, with up to 100% biallelic mutation efficiency in mouse embryos. Unlike BE3, HF2-BE2 could convert C to T on both the target and non-target strand, expanding the editing scope of base editors. Surprisingly, we found HF2-BE2 could also deaminate C that was proximal to the gRNA-binding region. Taken together, our work demonstrates the feasibility of generating point mutations in mouse by base editing, and underscores the need to carefully optimize base editing systems in order to eliminate proximal-site deamination.


Subject(s)
Animals , Humans , Mice , APOBEC-1 Deaminase , Genetics , Metabolism , Bacterial Proteins , Genetics , Metabolism , Base Sequence , CRISPR-Associated Protein 9 , CRISPR-Cas Systems , Cytidine , Genetics , Metabolism , Embryo Transfer , Embryo, Mammalian , Endonucleases , Genetics , Metabolism , Gene Editing , Methods , HEK293 Cells , High-Throughput Nucleotide Sequencing , Mice, Inbred C57BL , Microinjections , Plasmids , Chemistry , Metabolism , Point Mutation , Genetics , Metabolism , Thymidine , Genetics , Metabolism , Zygote , Metabolism , Transplantation
10.
Protein & Cell ; (12): 811-822, 2017.
Article in English | WPRIM | ID: wpr-756922

ABSTRACT

β-Thalassemia is a global health issue, caused by mutations in the HBB gene. Among these mutations, HBB -28 (A>G) mutations is one of the three most common mutations in China and Southeast Asia patients with β-thalassemia. Correcting this mutation in human embryos may prevent the disease being passed onto future generations and cure anemia. Here we report the first study using base editor (BE) system to correct disease mutant in human embryos. Firstly, we produced a 293T cell line with an exogenous HBB -28 (A>G) mutant fragment for gRNAs and targeting efficiency evaluation. Then we collected primary skin fibroblast cells from a β-thalassemia patient with HBB -28 (A>G) homozygous mutation. Data showed that base editor could precisely correct HBB -28 (A>G) mutation in the patient's primary cells. To model homozygous mutation disease embryos, we constructed nuclear transfer embryos by fusing the lymphocyte or skin fibroblast cells with enucleated in vitro matured (IVM) oocytes. Notably, the gene correction efficiency was over 23.0% in these embryos by base editor. Although these embryos were still mosaic, the percentage of repaired blastomeres was over 20.0%. In addition, we found that base editor variants, with narrowed deamination window, could promote G-to-A conversion at HBB -28 site precisely in human embryos. Collectively, this study demonstrated the feasibility of curing genetic disease in human somatic cells and embryos by base editor system.


Subject(s)
Female , Humans , APOBEC-1 Deaminase , Genetics , Metabolism , Base Sequence , Blastomeres , Cell Biology , Metabolism , CRISPR-Cas Systems , Embryo, Mammalian , Metabolism , Pathology , Fibroblasts , Metabolism , Pathology , Gene Editing , Methods , Gene Expression , HEK293 Cells , Heterozygote , Homozygote , Point Mutation , Primary Cell Culture , Promoter Regions, Genetic , Sequence Analysis, DNA , beta-Globins , Genetics , Metabolism , beta-Thalassemia , Genetics , Metabolism , Pathology , Therapeutics
11.
Chinese Journal of Information on Traditional Chinese Medicine ; (12): 56-59, 2016.
Article in Chinese | WPRIM | ID: wpr-483557

ABSTRACT

Objective To observe the mechanism of action of different extracts ofTangwang Mingmu Granules on high glucose induced VEGF and IL-1α gene and protein expressions in vascular endothelial cells.Methods Human umbilical vein endothelial cells EA.hy926 were divided into six groups: blank, high glucose,Tangwang Mingmu Granules, extract 1 (glycoside and flavonoid), extract 2 (organic acid and polysaccharides) and extract 3 (alkaloids) groups. High concentration glucose was used to establish the high glucose model in EA.hy926 cells. The expressions of VEGF and IL-1α mRNA were detected by semi-quantitative RT-PCR. The contents of VEGF and IL-1α protein were tested by ELISA.Results The gene expression and protein levels of VEGF and IL-1α were significantly up-regulated under the high glucose condition (P extract 3> extract 2.Conclusion The action intensity of glycosides and flavonoids, alkaloids, organic acids and polysaccharides on VEGF and IL-1α expression inTangwang Mingmu Granules weakens in sequence.

12.
Chinese Acupuncture & Moxibustion ; (12): 396-401, 2016.
Article in Chinese | WPRIM | ID: wpr-319948

ABSTRACT

<p><b>OBJECTIVE</b>To explore the rhythm regulatory mechanism of interleukin-6 (IL-6) in the process of moxibustion for rheumatoid arthritis (RA).</p><p><b>METHODS</b>A total of 144 Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, a moxibustion group, a sham operation group, an operation group, an operation+moxibustion group, 24 rats in each one. Each group was divided into 4 time points (0:00 am, 6:00' am, 12:00 am, 6:00 pm), 6 rats in each time point. The Light-Dark 12 : 12 was given in all rats for light-dark cycle. Except the blank group, rats in the remaining groups were treated with intracutaneous injection of freund's complete adjuvant at right-side foot to establish the model of RA. After the model establishment, bilateral adrenal, glands were removed in the operation group and operation + moxibustion group, while those in the sham operation group were not removed with identical operation procedure. Rats in the moxibustion group and operation + moxibustion group were treated with grain-sized moxibustion from 7:00 am to 9:00 am at "Shenshu" (BL 23) and "Zusanli" (ST 36) once everyday, 6 times were taken as one session and 3 sessions were required tatclly, while rats in the remaining groups received identical fixation without moxibustion. The general health state and foot volume of rats were measured before model establishment, after establishment and after treatment. After treatment, rats were sacrificed at each time point to collect the blood sample and measure the content of IL-6 by using enzymne-immunoassay method.</p><p><b>RESULTS</b>Compared with the blank group, the foot swelling in the model group was obviously increased (P<0. 05); the IL-6 maintained circadian rhythm (P<0. 05), but the peak phase had a backward trend, famplitude had an increased trend and the median was significantly lifted (P<0. 05). Compared with model group, !the foot swelling in the moxibustion group was obviously decreased (P<0. 05); the IL-6 maintained circadian. rhythm (P<0. 05), and the peak phase had a forward trend, amplitude had a decreased trend and the median was significantly reduced (P<0. 05). Compared with the moxibustion group, the foot swelling in the operation--moxibustion group was obviously increased (P < 0.05); the IL-6 maintained circadian rhythm (P < 0.5), but the peak phase moved forwrd, and the median was significantly elevated (P < 0.05).</p><p><b>CONCLUSION</b>The IL-6 in plasma maintains significant pathological circadian rhythm in RA rats; with the complete hypothalamic-pituitary-adrenal axis, moxibustion is likely to regulate the circadian rhythm of IL-6 to play an important role of anti-inflammatory effect in RA rats.</p>


Subject(s)
Animals , Female , Humans , Male , Rats , Acupuncture Points , Arthritis, Rheumatoid , Metabolism , Therapeutics , Circadian Rhythm , Disease Models, Animal , Hypothalamus , Metabolism , Interleukin-6 , Metabolism , Moxibustion , Pituitary-Adrenal System , Metabolism , Rats, Sprague-Dawley , Time Factors
13.
Journal of International Oncology ; (12): 371-373, 2015.
Article in Chinese | WPRIM | ID: wpr-467330

ABSTRACT

Diffuse intrinsic pontine glioma( DIPG)is a highly invasive tumor located in the pons (middle)of the brain stem. They are usually diagnosed during childhood and account for 10% -15% of primary brain tumors in children. DIPG has a very poor prognosis. Fewer than 10% of DIPG patients survive more than 2 years after diagnosis. The imaging manifestations of DIPG are typical,and biopsy is only performed in atypi-cal cases. The tissue specimens of newly diagnosed DIPG are very few and limit its molecular biological research. Recent advances in surgical and molecular-analytic techniques have increased the safety of biopsy which has already been used in many clinical trials step by step. The research of DIPG′s molecular pathogenesis and treatment is sure to achieve new breakthroughs.

14.
Protein & Cell ; (12): 363-372, 2015.
Article in English | WPRIM | ID: wpr-757583

ABSTRACT

Genome editing tools such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) have been widely used to modify genes in model systems including animal zygotes and human cells, and hold tremendous promise for both basic research and clinical applications. To date, a serious knowledge gap remains in our understanding of DNA repair mechanisms in human early embryos, and in the efficiency and potential off-target effects of using technologies such as CRISPR/Cas9 in human pre-implantation embryos. In this report, we used tripronuclear (3PN) zygotes to further investigate CRISPR/Cas9-mediated gene editing in human cells. We found that CRISPR/Cas9 could effectively cleave the endogenous β-globin gene (HBB). However, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic. Off-target cleavage was also apparent in these 3PN zygotes as revealed by the T7E1 assay and whole-exome sequencing. Furthermore, the endogenous delta-globin gene (HBD), which is homologous to HBB, competed with exogenous donor oligos to act as the repair template, leading to untoward mutations. Our data also indicated that repair of the HBB locus in these embryos occurred preferentially through the non-crossover HDR pathway. Taken together, our work highlights the pressing need to further improve the fidelity and specificity of the CRISPR/Cas9 platform, a prerequisite for any clinical applications of CRSIPR/Cas9-mediated editing.


Subject(s)
Humans , Blastocyst , CRISPR-Cas Systems , Hemoglobins, Abnormal , Genetics , Metabolism , Zygote
15.
Chinese Journal of Information on Traditional Chinese Medicine ; (12): 45-49, 2015.
Article in Chinese | WPRIM | ID: wpr-478669

ABSTRACT

ObjectiveTo observe the effect of different extracts ofTangwang Mingmu Granule on hypoxia induced gene expressions in vascular endothelial cells.Methods COCl2 intervention cells were used to copy hypoxia models. Human umbilical vein endothelial cells EA. Hy926 were divided into blank group, hypoxia model group,Tangwang Mingmu Granule group, extract 1 (glycosides and flavonoids) group, extract 2 (orgain acids and polysaccharides) group and extract 3 (alkaloids) group. The changes in gene expressions of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α mRNA were detected by semi-quantitative RT-PCR.ResultsThe gene expression levels of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α were significantly up-regulated under the hypoxic condition (Palkaloids>organic acids and polysaccharides inTangwang Mingmu Granule.

16.
Chinese Pharmacological Bulletin ; (12): 1743-1747, 2014.
Article in Chinese | WPRIM | ID: wpr-458712

ABSTRACT

Aim To explore the role of ERO1 αand its DNA methylation in homocysteine (Hcy)-induced in-hibition of hepatocytes proliferation.Methods The hepatocytes stimulated with 0 μmol·L -1 Hcy were set as the normal group (NC group)and the hepatocytes stimulated with 1 00 μmol·L -1 Hcy as the experimen-tal group (Hcy group).Methyl thiazolyl tetrazolium (MTT)reduction assay was used to reflect the prolifer-ation of the hepatocytes;qRT-PCR and Western blot were used to detect the mRNA and protein levels of ERO1 α;the expression of green fluorescence protein was observed in hepatocytes after the recombinant plas-mid of ERO1 α was constructed,which was used to confirm if the recombinant plasmid into hepatocytes was successful,then the mRNA and protein levels of ERO1 αwere assayed and the proliferation of the hepa-tocytes was also detected;ntMSP was used to detect the change of ERO1 αDNA methylation.Results The mRNA and protein levels of ERO1 αwere decreased in Hcy group compared with NC group,and the prolifera-tion activity of hepatocytes in Hcy group was de-creased.Sequencing result showed that the recombi-nant plasmid of ERO1 αwas constructed successfully. QRT-PCR and Western blot revealed that ERO1 αwas overexpressed. The result of MTT suggested that ERO1 αoverexpression restored hepatocyte proliferation inhibited by Hcy.Hcy caused ERO1 αDNA hyperm-ethylation.Conclusions Hcy inhibits hepatocyte pro-liferation by downregulating the expression of ERO1 α, and methylation of ERO1 αpromoter may play a role in this process.

17.
Chinese Pharmacological Bulletin ; (12): 1287-1292, 2014.
Article in Chinese | WPRIM | ID: wpr-456609

ABSTRACT

Aim To investigate the possible mecha-nisms of the levels of NO decrease induced apoptosis in human placental trophoblast cells. Methods Human placental trophoblast cells ( HTR-8 ) were cultured in 5 ml DMEM-F12 culture medium with 37℃ 5% CO2 . Then, the old culture medium was discarded and re-placed with 10,100,500,1 000 μmol·L-1 L-NAME, and the group without L-NAME was set as the control group, cultured for 48h. The effects of L-NAME on the survival of cells were detected by methylthiazolyldiphe-nyl tetrazolium bromide ( MTT); the content of NO in cells was tested by nitrate reductive enzymatic;trans-mission electron microscopy, flow cytometry analysis and Annexin-V FITC dyeing were used to test the effects of L-NAME on apoptosis in HTR-8 cells;restore Fe3+ colorimetric assay was applied for detection of to-tal antioxidant capacity ( T-AOC ) , xanthine oxidase for detection of superoxide dismutase ( SOD) activity, and thiobarbituric acid colorimetry for determination of content of MDA. Results Compared with the control group, the survival rate of HTR-8 cells and the levels of NO in 100,500,1 000 μmol·L-1 L-NAME group were significantly reduced(P<0.05,P<0.01). Flow analysis and Annexin-V FITC staining showed that L-NAME could induce cell apoptosis in a dose-dependent manner. The number of cell apoptosis was negatively correlated with the content of NO ( r = -0.5210 ) in HTR-8 cells. Transmission electron microscopy results showed that compared with the control group, the ex-perimental group's cell nucleus shape was irregular, nuclear pyknosis in irregular shape, the chromatin ag-glutination or side the collection, mitochondrial swell-ing or enrichment, crest fracture or dissolved, even vanished, forming the vacuole, especially in 100 μmol ·L-1 L-NAME group, the apoptotic bodies obviously appeared. At the same time, T-AOC, SOD levels in HTR-8 cells decreased ( P <0.05 ) , and the MDA content increased ( P<0.05 ) . The number of cell ap-optosis was negatively correlated with the level of T-AOC ( r= -0.3212 ) , SOD ( r= -0.2779 ) in HTR-8 cells , while positively correlated with the content of MDA(r=0.2807). Conclusion Oxidative stress may play an important role in the levels of NO decrease in-duced apoptosis in human placental trophoblast cells.

18.
Clinical Medicine of China ; (12): 339-342, 2013.
Article in Chinese | WPRIM | ID: wpr-432051

ABSTRACT

Objective To investigate the differences of related clinicopathologic features in colorectal cancer (CRC) patients with or without Type 2 Diabetes Mellitus (T2DM).Methods Using case-control studies,retrospective analysis of colorectal cancer of 60 patients with colorectal cancer hospitalization and 32 cases of T2DM were taken.The clinical data,histological grade,tumor size,clinical stage and lymph node metastasis pathology indicators conditions of the hospitalized CRC patients with or without type 2 diabetes were compared.Results The average age of the team with hospitalized CRC patients combined with type 2 diabetes is (64.90 ±8.87),fasting blood-glucose is (8.33 ±4.66) mmol/L.BMI and TG are (25.77 ±3.80) kg/m2 and (1.71 ±0.85) mmol/L which were all higher((54.70 ± 11.62),(5.85 ±0.88) mmol/L,(23.57 ±3.64)kg/m2,(1.33 ± 0.83) mmol/L) than patients without DM.There were significant differences between two groups (t =4.324,t =2.982,t =2.728,t =2.045,respectively,P < 0.05).However,total cholesterol,high density lipoprotein-cholesterol and low density lipoprotein-cholesterol had no significantly differences with those patients without DM (P > 0.05).There were no significant differences of clinicopathologic features between CRC patients with and without DM (P > 0.05).Conclusion There is a certain correlation between CRC and DM,DM increases the risk of colorectal cancer so diabetes patients should be early screened for colorectal,with a view to early detection,treatment and improve the prognosis.

19.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 1-2, 2012.
Article in Chinese | WPRIM | ID: wpr-417825

ABSTRACT

ObjectiveTo investigate the control rate and influence factors of glycolated hemoglobin A1C(HbA1c) of type 2 diabetes in Baoding city community.MethodsA cluster-randomized study was conducted and three communities were selected randomly.The study involved all of people aged 45 years old and above in the three communities.The type 2 diabetic patients diagnosed before,were recorded through the cross section study.Then the questionnaire was finished.All diabetic plasma HbA1c was determined by HPLC method.ResultsEighty-seven patients with HbA1c≤7% were only 18.6% in all diabetic patients.As the plasma level of HbA1c increased,occurrences of macroangiopathy and microangiopathy were both increased,by the trend test.Logistic regression analysis showed that therapeutic measures and knowledge about diabetes were main influence factors of achieving the hemoglobin Alc target of <7% in type 2 diabetes.ConclusionThere were low HbA1c control in diabetic patients of Baoding community,and knowing diabetes well and receiving insulin treatment might decrease HbA1c level apparently.

20.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 2252-2253, 2012.
Article in Chinese | WPRIM | ID: wpr-427835

ABSTRACT

Objective To analyze the cause of acute brain swelling,brain encephalocele in brain traumatic injury craniotomy,and the reason,diagnosis and treatment methods of progressive epidural hematoma.Methods The clinical data of 381 patients with brain traumatic injury craniotomy were retrospetively analyzed.Of 318 patients,27cases had progressive epidural hematoma during operation.Results 9 cases died because of functional failure of brain stem,the other 18 cases were all clinically cured.Conclusion If acute brain swelling and encephalocele occured during operation,the cause should be found quickly,especially when there was skull fracture or epidural hematoma on the contralateral brain,the head CT and operation should be taken immediately,and the bulging brain tissue should not be removed blindly,and the skull should not be forced to close,otherwise the prognosis maybe poor.

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