1.Expression pattern of genes involved in tropane alkaloids biosynthesis and tropane alkaloids accumulation in Atropa belladonna.
Wei QIANG ; Ya-Xiong WANG ; Qiao-Zhuo ZHANG ; Jin-Di LI ; Ke XIA ; Neng-Biao WU ; Zhi-Hua LIAO
China Journal of Chinese Materia Medica 2014;39(1):52-58
Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.
Alkaloids
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biosynthesis
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genetics
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metabolism
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Atropa belladonna
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genetics
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metabolism
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Gene Expression Regulation, Plant
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genetics
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Hyoscyamine
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genetics
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metabolism
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Plants, Medicinal
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genetics
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metabolism
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Scopolamine Hydrobromide
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metabolism
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Tropanes
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metabolism
2.Effect of Acupuncture on Histology of Thymus and Expressions of ANP and NPR-A in Anxiety Rats.
Wen-qin HUANG ; Da-neng WEI ; Qi-zhi ZHOU ; Shu-guang YU ; Ding-jun CAI ; Wei JING-JIN ; Xia XIAO ; Hong YAO ; Guang-chun XIE ; Ming-ji YANG ; Zhen-dong ZHONG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(2):216-221
OBJECTIVETo explore possible mechanism of electroacupuncture (EA) for regulating immune function in anxiety disorder (AD) rats by observing the effect of acupuncture on the histology of thymus and expressions of atrial natriuretic peptide (ANP) and natriuretic peptide receptor type A (NPR- A) in thymus.
METHODSTotally 34 SD healthy rats were randomly divided into the blank control group (n = 10), the model group (n = 12), the EA group (n = 12). Anxiety model was established in rats of the model group and the EA group by using chronic unpredictable stress (CUS) stimulation. EA (15/25 Hz) at Neiguan (PC6) and Shenmen (HT7) was performed in the EA group, with 15-min needle retaining, once every other day, 15 days in total. Needle was fixed at same acupoints for 15 min without electric stimulus in the other two groups. Anxiety-like behavior was measured by elevated plus-maze (EPM) test. Pathological changes of thymus tissue were observed by optical microscope. Expressions of ANP and NPR-A in thymus were measured by immunohistochemical assay.
RESULTSThe thymus tissue in the model group was severely atrophied, with unclear structure of thymic lobules, unclear margin of thymic medulla, loosely arranged lymphocytes ,and obviously enlarged volume of thymic corpuscle. The thymus tissue in the EA group was mildly atrophied, with existent structure of thymic lobules, clear margin of thymic medulla, densely arranged lymphocytes in cortical region, and widened medullary area. Com- pared with the blank control group, the percentage of open-arms entries (OE%) in the total QE times ob- viously decreased in the model group (P < 0.05), ANP expression obviously increased (P < 0.05), and NPR-A expression obviously decreased (P < 0.01). Compared with the model group, OE% was obviously elevated (P < 0.05), ANP expression obviously decreased (P < 0.05), and NPR-A expression obviously increased (P < 0.01) in the EA group.
CONCLUSIONEA not only could reduce anxiety of rats, but also could improve chronic stress induced thymus injury through intervening synthesis and secretion of ANP, as well as the expression of NPR-A (a specific receptor of ANP).
Acupuncture Points ; Animals ; Anxiety Disorders ; therapy ; Atrial Natriuretic Factor ; metabolism ; Electroacupuncture ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Atrial Natriuretic Factor ; metabolism ; Thymus Gland ; pathology
3.Azoospermia factor microdeletion on Y chromosome in patients with idiopathic azoospermia or severe oligozoospermia.
Lan-Ping HU ; Neng-Hui LIU ; Qian PAN ; De-Sheng LIANG ; Zhi-Gao LONG ; Hao HU ; Hai-Yan ZHU ; He-Ping DAI ; Fang CAI ; Ling-Qian WU ; Kun XIA ; Jia-Hui XIA
Journal of Central South University(Medical Sciences) 2007;32(2):241-245
OBJECTIVE:
To investigate the correlation between male infertility and Y chromosome microdeletions of azoospermia factor (AZF) regions, and to establish a reliable genetic diagnosis in idiopathic infertile male patients with azoospermia or severe oligozoospermia.
METHODS:
Multiplex PCR amplification of 6 sequence-tagged sites in AZF regions of the Y chromosome was examined among 100 normal karyotype male patients with azoospermia or oligozoospermia.
RESULTS:
Four patients (4%) had Y chromosome microdeletions, the microdeletions of 3 patients were idiopathic azoospermic and those of the other 1 patient were secretory azoospermia.
CONCLUSION
The PCR-based Y chromosome microdeletion screening is simple and effective in the diagnosis of patients with severe male infertility. Microdeletion of Y chromosome is one of the major causes of severe dyszooospermia.
Adult
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Azoospermia
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genetics
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Chromosome Deletion
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Chromosomes, Human, Y
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genetics
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Genetic Loci
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Humans
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Infertility, Male
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diagnosis
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genetics
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Karyotyping
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Male
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Oligospermia
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genetics
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Seminal Plasma Proteins
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genetics
4.Application of Virtual Anatomy Technology in Postmortem Examination of Medical Dispute Cases.
Xin JIN ; Jian Xin ZHAO ; Yi YAO ; Jun Jie HUANG ; Feng ZHANG ; Xing Biao LI ; Guang Hua YE ; Yan Yan FAN ; Ding Pin HUANG ; Neng Zhi XIA ; Dong Hua ZOU ; Ning Guo LIU ; Lin Sheng YU
Journal of Forensic Medicine 2020;36(1):72-76
Objective To discuss the application value of CT scanning technology in cause of death determination of medical dispute cases. Methods From July 2017 to December 2018, postmortem CT imaging data of 12 medical dispute cases were collected. CT imaging diagnosis results and anatomy findings as well as differences between antemortem and postmortem CT diagnosis were compared. The advantages and disadvantages of CT routine tests of the cadavers in terms of the diagnosis of disease and damage were analyzed. Results The comparison between CT imaging diagnosis and anatomical findings showed that CT scans had advantages in the diagnosis of disease and damage with large differences in density changes, such as atelectasis, pneumonia, calcification, fracture and hemorrhage, etc. The comparison of CT diagnosis in antemortem and postmortem examination showed that the cadavers of medical dispute cases were well preserved and that postmortem CT scan was meaningful for the diagnosis of antemortem diseases. Conclusion Virtual anatomy technology has a relatively high application value in postmortem examination of medical dispute cases. It can provide effective information for the appraisers before the autopsy and can also provide a reference for cause of death analysis when the anatomy cannot be performed.
Autopsy
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Cadaver
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Dissent and Disputes
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Humans
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Postmortem Changes
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Tomography, X-Ray Computed