1.The diversity of endophytic fungi in endangered medicinal material Daphnes Cortex as determined by high-throughput sequencing technology
Yuan HE ; Qi-rui MU ; Zhen-fang BAI ; Dan JIANG ; Guang-xi REN ; Chun-sheng LIU
Acta Pharmaceutica Sinica 2021;56(3):872-878
In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%.
2.The degradation of housekeeping mRNA in dead rats by real-time RT-PCR.
Guang-mu REN ; Ji LIU ; Ying-yuan WANG
Journal of Forensic Medicine 2009;25(1):33-36
OBJECTIVE:
To explore the feasibility of real-time RT-PCR of housekeeping mRNA degradation in dead rats in order to seek new suitable techniques for post-mortem interval (PMI).
METHODS:
The levels of GAPDH mRNA and beta-actin mRNA in the brain and spleen of the rats were measured at different times after death by the SYBR Green I real-time RT-PCR. The relative mRNA level was indicated with the Ct value, then the relationship between the Ct value and PMI were analyzed and the corresponding regression equation was obtained.
RESULTS:
The Ct values of GAPDH mRNA and beta-actin mRNA by SYBR Green I real time RT-PCR correlated well with the PMI.
CONCLUSION
The SYBR Green I real-time RT-PCR is a reliable technique for studying mRNA degradation. Adoption of the housekeeping genes eliminates systematical errors of other genes which have individual difference. As an objective and dynamic indication, Ct value has a good correlation with PMI, and could be applied for estimating PMI, especially in late period.
Actins/genetics*
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Animals
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Glyceraldehyde-3-Phosphate Dehydrogenases/genetics*
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Postmortem Changes
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RNA Stability/genetics*
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RNA, Messenger/metabolism*
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Rats
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Rats, Sprague-Dawley
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Reverse Transcriptase Polymerase Chain Reaction/methods*
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Time Factors
3.Lethal anaphylactic shock model induced by human mixed serum in guinea pigs.
Guang-Mu REN ; Ji-Wei BAI ; Cai-Rong GAO
Journal of Forensic Medicine 2005;21(3):169-170
OBJECTIVE:
To establish an anaphylactic shock model induced by human mixed serum in guinea pigs.
METHODS:
Eighteen guinea pigs were divided into two groups: sensitized and control, The sensitized group were immunized intracutaneously with human mixed serum and then induced by endocardiac injection after 3 weeks.
RESULTS:
Symptoms of anaphylactic shock appeared in the sensitized group. The level of serum IgE were increased in the sensitized group significantly.
CONCLUSIONS
An animal model of anaphylactic shock wer established successfully. It provide a tool for both forensic study and anaphylactic shock therapy.
Allergens
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Anaphylaxis/immunology*
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Animals
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Death, Sudden
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Disease Models, Animal
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Female
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Forensic Medicine
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Guinea Pigs
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Humans
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Immunoglobulin G/blood*
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Male
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Radioimmunoassay
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Random Allocation
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Serum/immunology*
4.The correlation between the DNA content in human thyroid follicular epithelial cells and the postmortem interval.
Guang-Mu REN ; Cai-Liang JING ; Ji LIU ; Ya-Fang WANG ; Ying-Yuan WANG
Journal of Forensic Medicine 2007;23(2):92-96
OBJECTIVE:
To study the relationship between the DNA content in follicular epithelial cells of the human thyroid and postmortem interval (PMI).
METHODS:
Changes of the DNA content in thyroid follicular epithelial cells at different PMI were determined by Methyl Green-Pyronin (MGP) stain combined with an image analysis technique.
RESULTS:
The average DNA content in the thyroid follicular epithelial continued to decrease with increased PMI. The coefficient of determination for DNA content with average gray, aimed area, aimed area ratio and positive index in follicular epithelial cells were 0.960, 0.987, 0.988 and 0.990, respectively.
CONCLUSION
There is an apparent correlation between the average DNA content of follicular epithelial cells and the PMI. MGP stain combined with an image analysis technique seems to be a useful means to study the degradation of DNA in thyroid follicular epithelial cells.
Adult
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Cadaver
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Cell Nucleus/metabolism*
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DNA/metabolism*
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Epithelium/metabolism*
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Female
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Forensic Pathology
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Humans
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Image Processing, Computer-Assisted/methods*
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Male
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Middle Aged
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Postmortem Changes
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Rosaniline Dyes
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Staining and Labeling/methods*
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Thyroid Gland/metabolism*
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Time Factors
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Young Adult
5.Relationship between Radiological Stages and Prognoses of Pneumocystis Pneumonia in Non-AIDS Immunocompromised Patients.
Xiang-Dong MU ; Peng JIA ; Li GAO ; Li SU ; Cheng ZHANG ; Ren-Gui WANG ; Guang-Fa WANG
Chinese Medical Journal 2016;129(17):2020-2025
BACKGROUNDAlthough radiological features of pneumocystis pneumonia (PCP) in non-Acquired Immune Deficiency Syndrome (AIDS) immunocompromised patients have been reported by other authors, there were no studies on the radiological stages of PCP previously. This study aimed to elucidate the radiological stages and prognoses of PCP in non-AIDS immunocompromised patients.
METHODSRetrospective analysis of radiological manifestations and prognoses of 105 non-AIDS PCP immunocompromised patients from August 2009 to April 2016 was conducted. Chest radiograph was divided into three stages: early stage (normal or nearly normal chest radiograph), mid stage (bilateral pulmonary infiltrates), and late stage (bilateral pulmonary consolidations); chest high-resolution computed tomography (HRCT) was also divided into three stages: early stage (bilateral diffuse ground-glass opacity [GGO]), mid stage (bilateral diffuse GGO and patchy consolidations), and late stage (bilateral diffuse consolidations).
RESULTSThe case fatality rate (CFR) of all patients was 34.3% (36/105), all of them took routine chest X-ray (CXR), and 84 underwent chest CT examinations. According to the CXR most near the beginning of anti-PCP therapy, 18 cases were at early stage and CFR was 0 (0/18, P< 0.01), 50 cases were at mid stage and CFR was 28.0% (14/50, P> 0.05), and 37 cases were at late stage and CFR was 59.5% (22/37, P< 0.01). According to the chest HRCT most near the beginning of anti-PCP therapy, 40 cases were at early stage and CFR was 20.0% (8/40, P> 0.05), 34 cases were at mid stage and CFR was 47.1% (16/34, P> 0.05), and 10 cases were at late stage and CFR was 80.0% (8/10, P< 0.05); barotrauma, including pneumothorax, pneumomediastinum, and pneumohypoderma, was found in 18 cases and the CFR was 77.8% (14/18, P< 0.01).
CONCLUSIONSBased on the radiological manifestations, the course of PCP in non-AIDS immunocompromised patients can be divided into three stages: early stage, mid stage, and late stage. The prognoses of patients treated at early stage are good, and those at late stage are poor. Furthermore, the CFR of patients with barotrauma is high.
Acquired Immunodeficiency Syndrome ; complications ; mortality ; pathology ; Adult ; Female ; Humans ; Immunocompromised Host ; Male ; Middle Aged ; Pneumonia, Pneumocystis ; diagnosis ; mortality ; pathology ; Prognosis ; Retrospective Studies ; Tomography, X-Ray Computed
6.Correlation between chemical composition,ecological factors and soil factors of Chinese herbal medicine Daphnes Cortex.
Qi-Rui MU ; Dan JIANG ; Yuan HE ; Lu GENG ; Guang-Xi REN ; Zhen-Fang BAI ; Xu ZHANG ; Zhong-Yi ZHANG ; Chun-Sheng LIU
China Journal of Chinese Materia Medica 2020;45(5):1059-1063
In this paper, the correlation between the chemical constituents of Chinese herbal medicines Daphnes Cortex and the ecological factors and soil factors was studied, which provided a reference for the selection of suitable areas for artificial cultivation of Daphne giraldii and wild tending. The geographic information system(GIS) was applied to obtain the ecological factor information of 23 collection sites of Daphnes Cortex, and the soil factor information was determined by the standard procedure in the soil test standard manual. Combining the information of 93 chemical constituents of Daphnes Cortex in 23 collection sites the correlation between components and ecological factors and soil factors was analyzed by statistical methods. The correlation analysis showed that the longitude, annual average rainfall, annual sunshine intensity, annual average temperature in the ecological factors, soil type, effective copper and pH value were the dominant factors affecting the chemical composition of Daphnes Cortex.
China
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Copper
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Daphne/chemistry*
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Drugs, Chinese Herbal
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Geographic Information Systems
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Hydrogen-Ion Concentration
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Plants, Medicinal/chemistry*
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Rain
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Soil/chemistry*
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Sunlight
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Temperature
7.Mechanisms of sorafenib induced NB4 cell apoptosis.
Yun-Jie ZHANG ; Xin LIU ; Yan-Ping SONG ; Gang-Can LI ; Nai-Cen ZHOU ; Hao WANG ; Qi-Xia WANG ; Jia XIE ; Guang LI ; Jing-Jing REN ; Fei GAO ; Xiao-Bo ZHANG ; Jin-Qian DAI ; Lu WANG ; Jiao MU
Journal of Experimental Hematology 2015;23(1):77-82
OBJECTIVETo investigate the effects of sorafenib on human acute promyelocytic leukemia cell NB4 and its mechanism.
METHODSThe human acute promyelocytic leukemia cell NB4 was treated with different concentrations (0, 1.5, 3, 6 and 12 µmol/L) of sorafenib, the proliferation inhibitory rate of NB4 cells was assayed by MTT, the apoptosis of NB4 was determined with flow-cytomatry after treatment; after extraction of total protein, the Western blot was performed to determine the expressions of apoptosis-relatived molecules Caspase-3, Caspase-8 and MCL-1. The mRNA expressions of Caspase-3, Caspase-8 and MCL-1 were determined by RT-PCR.
RESULTSAs compared with the control group, the proliferation of NB4 significantly decreased after treatment with different concentrations of sorafenib. The sorafenib significantly induced the apopotosis of NB4 cells in time- and dose-dependent manners. Furthermore, sorafenib treatment resulted in the obvious increase of the Caspase-3 and Caspase-8 protein and mRNA expressions, and down-regulated the MCL-1 protein and mRNA expressions in NB4 cells.
CONCLUSIONSorafenib can inhibit proliferation and induce apopotosis of human acute promyelocytic leukemia cell NB4 through the expression of Caspase-3 and Caspase-8, and down-regulation of the expression of MCL-1.
Antineoplastic Agents ; Apoptosis ; Caspase 3 ; Caspase 8 ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; T-Lymphocytes, Helper-Inducer