1.Enzyme-linked Immunosorbent Assay in Quality Control of Chinese Medicines: A Review
Yuansheng XU ; Jiao LIU ; Tian ZHANG ; Yuyang ZHAO ; Hui TIAN ; Tiegui NAN ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):12-20
In the quality control of Chinese medicine, the detection of active components and toxic and harmful components are two important links. Although conventional methods such as high performance liquid chromatography and liquid chromatography-mass spectrometry can accurately quantify the above substances, they have shortcomings such as complicated operation, high costs, inability of detection at any time, difficult detection of insoluble and macromolecular substances. Enzyme-linked immunosorbent assay (ELISA) can adsorb antigens or antibodies on the surface of solid carriers and realize qualitative or quantitative analysis of targets by using the specific reactions of antigens and antibodies. This method is praised for the simple operation, high sensitivity, strong specificity, simple requirements for experimental equipment, a wide application range, and low costs. In recent years, ELISA has been widely used in the quality control of Chinese medicine, especially in the content determination of mycotoxins represented by aflatoxin and the qualitative and quantitative analysis of active components. ELISA plays an increasingly important role with its unique advantages, providing new methods and ideas for the rapid quality examination of large quantities of Chinese medicines. This paper reviews the research progress in ELISA for the quality control of Chinese medicine in recent years and prospects its technical development and application prospects, aiming to provide reference and research ideas for further using this method to ensure the quality, safety, and controllability of Chinese medicine.
2.Identification of Scolopendra Dispensing Granules by Allele-specific PCR
Yuansheng XU ; Li HU ; Chao JIANG ; Yuyang ZHAO ; Tianyun CHEN ; Hui ZHANG ; Hui TIAN ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):48-54
ObjectiveTo establish an allele-specific polymerase chain reaction (PCR) method for identifying Scolopendra dispensing granules, so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3(COX-3) gene sequence of Scolopendra, and the single nucleotide polymorphism (SNP) loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature, cycles, Taq enzymes, DNA template amount, PCR instruments, and primer concentrations, and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix, 0.4 μL forward primer (10 μmol·L-1), 0.4 μL reverse primer (10 μmol·L-1), 2.5 μL DNA template, and 9.2 μL sterile double distilled water. PCR parameters: Pre-denaturation at 94 ℃ for 3 min, 30 cycles (94 ℃ for 20 s, 62 ℃ for 20 s, 72 ℃ for 45 s), and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above, the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials, decoction pieces, and standard decoction freeze-dried powder of Scolopendra, as well as the intermediates and final products of Scolopendra dispensing granules, which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.
3.A modal study of the effect of adjusting density exposure steps on image quality and radiation dose in digital mammography
Xia XU ; Zhifeng WU ; Yuansheng ZHANG ; Yue TIAN ; Na ZHANG ; Ziquan GUO
Chinese Journal of Radiology 2023;57(4):410-415
Objective:To investigate the effect of adjusting density exposure steps on image quality and radiation dose in digital mammography.Methods:Using the automatic exposure control (AEC) mode of the digital mammography machine, five different gland thicknesses of 4.3, 5.3, 6.3, 7.3, and 8.3 cm were simulated by attaching 0, 1, 2, 3, and 4 PMMA plexiglass plates under the RMI-156 modal body, and the density exposure steps were adjusted to -3, -2, -1, 0, 1, 2, 3, and 4 for each thickness. The target/filter combination, tube voltage, tube current, incident body surface dose (ESD), incident surface air kerma (ESAK), half-value layer (HVL) and the average glandular dose displayed by the device (displayed AGD) were recorded at each step and thickness, and the signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), figure of merit (FOM) and the calculated average glandular dose (calculated AGD) were calculated. Then, the display effects of simulated fibers, simulated calcifications and simulated masses within the modal body were scored subjectively, and the changes in image quality and radiation dose at different steps were analyzed, and the relationships between ESD and ESAK, displayed AGD and calculated AGD, and displayed AGD/ESD and calculated AGD/ESAK were analyzed. A linear fit was used for the steps with SNR, CNR, and FOM, and an exponential function curve fit was used for the steps with mAs, ESAK, and calculated AGD. The differences between ESD and ESAK, displayed AGD and calculated AGD, and displayed AGD/ESD and calculated AGD/ESAK were analyzed by paired-samples t test. Results:The CNR and SNR of mammographic images rose and fell by about 8% with each increase or decrease of one step. The scores of image simulated fibers, simulated calcifications, and simulated masses showed an overall upward trend with increasing steps, but there were still cases where the scores decreased with increasing grades. FOM varied from 97% to 104% at each grade with little variability. ESD, ESAK, displayed AGD, and calculated AGD, which could measure radiation dose, showed an exponential trend of increasing function with increasing steps, with a variation of about 63% to 165%. There were statistically significant differences ( t=-9.61, P=0.001) between ESD (15.14±10.08) and ESAK (16.66±11.07). However, there were no statistically significant differences ( t=1.20, P=0.240) between displayed AGD and calculated AGD, which were 3.66±2.18 and 3.61±1.99, respectively. Conclusions:The adjustment of density exposure steps can make the image quality change linearly and the radiation dose change exponentially with increasing speed, and the mode and magnitude of the adjustment are appropriately stable with high application value.
4.Analyses on horizontal scientific research projects from 2015 to 2019 by provincial Center for Disease Control and Prevention in China
Xi YANG ; Meng WANG ; Xiaojie LI ; Yudan SONG ; Liang CHEN ; Yuansheng CHEN ; Jing XU ; Yu QIN ; Yanhua HOU ; Jichun WANG
Chinese Journal of Medical Science Research Management 2022;35(1):36-41
Objective:To analyze the horizontal scientific research projects from 2015 to 2019 by the provincial Centers for Disease Control and Prevention (CDC) in China, and to compare the regional differences, in order to provide the suggestion on the scientific management of CDC.Methods:The horizontal scientific research projects from 2015 to 2019 were retrospectively analyzed by questionnaire survey. Multiple linear regression models were adopted to examine the trend, and variance analyses were used to test the differences in horizontal scientific research projects among the Eastern, Central, and Western regions.Results:From 2015 to 2019, provincial CDC have received RMB 124.3 million of horizontal scientific research project funds totally, of which 51.9% were funded by enterprises, and 86.9% were undertaken by provincial CDC themselves. There were no statistical significance in the change of research project funds obtained by provincial CDC ( F=0.46, P = 0.764) during this period.The number of horizontal scientific research projects undertook or participated by provincial CDC in the Eastern region were more than that of the Central and Western region ( F = 5.85, P = 0.004; F = 5.03, P = 0.008). Conclusions:The horizontal scientific research projects obtained by the provincial CDC remained stable in recent years while distribution was unbalanced in the region areas. It is suggested to innovate the management mode of scientific research projects with strengthening the trans-agency, trans-department and trans-regional cooperation.
5.Therapeutic effect of anti-inflammatory tripeptide KdPT on ophthalmoxerosis in mice
Hua WANG ; Yuansheng XU ; Yan SONG
Journal of China Pharmaceutical University 2021;52(1):100-103
This study was designed to investigate the therapeutic effect of anti-inflammatory tripeptide KdPT on ophthalmoxerosis. Male BALB/c mice, 8-week old, were treated with 0.2% benzalkonium chloride solution to establish the ophthalmoxerosis model. Four weeks after modeling, the mice were randomly divided into control group, positive group and the low, medium, high dose groups of KdPT. Each group was given normal saline, artificial tears and 1, 10, 100 μg/mL KdPT, respectively. After 3, 5, 7, 10 and 14 days of treatment, the morphology of the eye surface was observed, and the fluorescein sodium staining score was performed. The amount of tear secretion was measured by phenol red cotton thread and the right corneas were taken out for histopathological analysis after 14 days of treatment. Data showed that there was no significant abnormality in general state and the weight of mice in each group at each time point of treatment. After 14 days of treatment, KdPT can promote the secretion of tear, repair the damaged corneal epithelium, and showed a significant therapeutic effect on ophthalmoxerosis in mice. Based on the data, it is possible for KdPT to be developed as a novel drug for ophthalmoxerosis.
6.Progress of research on disease-modifying osteoarthritis drugs
SU Boya ; XU Yuansheng ; WANG Hua ; TANG Yuqing ; ZHANG Shiqun ; SONG Yan
Journal of China Pharmaceutical University 2021;52(2):253-260
Osteoarthritis (OA) is a common chronic joint disease,whose main pathological changes are the degeneration of articular cartilage and secondary bone hyperplasia.The limitation of current treatment methods including pain relief and joint replacement surgery is that they cannot fundamentally improve the damage of articular cartilage.The emergence of disease-modifying osteoarthritis drugs (DMOAD) may break the above limitations.They fundamentally inhibit the structural degeneration of articular cartilage by participating in the regulation of cartilage metabolic balance, regulation of subchondral bone remodeling,and control of local inflammation.Thereby,OA patients will get symptom improvement including pain relief and joint function restoration,delay the artificial joint replacement surgery, and improve the quality of life. There are still no DMOAD drugs widely available on the market worldwide.This paper reviews the background of R&D,the classification of mechanisms of action and research progress of representative drugs under different inechanisms so as to provide reference for future research.
7.Morphological analysis of meibomian glands in patients with meibomian gland cyst under in vivo confocal microscope
Jianhao CAI ; Cangeng XU ; Lingling ZHOU ; Zifeng ZHAO ; Zeyi LI ; Yuansheng ZHOU
Chinese Journal of Experimental Ophthalmology 2021;39(2):139-143
Objective:To analyze the morphological changes and features of meibomian gland in patients with meibomian gland cyst under in vivo confocal microscope (IVCM). Methods:A cross-sectional study was performed.A total of 34 patients (34 eyes) with meibomian gland cysts and 18 control subjects (18 eyes) in the outpatient department without meibomian gland cysts treated in Shantou International Eye Center from September 2018 to April 2019 were included into the meibomian gland cyst group and control group accordingly.All the subjects underwent routine ophthalmologic examination and IVCM examination.IVCM test indicators included the opening area of meibomian gland, the longest diameter and the shortest diameter of meibomian gland opening, the morphology of glandular tube and acinus adjacent to the meibomian gland opening.The measurement indexes of the meibomian gland cyst group and the control group were compared and analyzed.This study followed the Declaration of Helsinki and was approved by the Ethics Committee of Joint Shantou International Eye Center of Shantou University and The Chinese University of Hong Kong (No.EC20171103[6]-P01). Written informed consent was obtained from each patient before examination.Results:The opening of the meibomian glands of the 34 subjects in the meibomian gland cyst group were all enlarged irregularly with smooth boundaries, and emboli in the openings were observed in 70.59% (24/34) of patients.The longest diameter, the shortest diameter and the area of meibomian gland openings were (148.12±70.16)μm, (114.77±52.38)μm and 9 239.11(5 506.96, 24 111.36)μm 2 in the meibomian gland cyst group, respectively, while (59.35±16.78)μm, (41.98±11.77)μm and 2 094.19 (1 432.28, 2 945.65)μm 2 in the control group, respectively.Compared with the control group, the longest diameter and shortest diameter in the meibomian cyst group were longer, and the area of meibomian gland openings in the meibomian cyst group was larger, and the differences were statistically significant (all at P<0.01). Adjacent to the opening, there was cystic dilation of glandular tube containing accumulated secretion of different characteristics detected in the 61.76% (21/34) of patients in the meibomian gland cyst group, and the dilated glandular tubes were with flat edges.The boundaries between the dilated glandular tubes and surrounding acini were clear. Conclusions:In vivo confocal microscope can detect the morphological changes of meibomian glands in patients with meibomian gland cyst, including enlarged opening with embolus, cystic dilation of glandular tube with clear boundary and accumulated secretion.
8.IC4R-2.0:Rice Genome Reannotation Using Massive RNA-seq Data
Sang JIAN ; Zou DONG ; Wang ZHENNAN ; Wang FAN ; Zhang YUANSHENG ; Xia LIN ; Li ZHAOHUA ; Ma LINA ; Li MENGWEI ; Xu BINGXIANG ; Liu XIAONAN ; Wu SHUANGYANG ; Liu LIN ; Niu GUANGYI ; Li MAN ; Luo YINGFENG ; Hu SONGNIAN ; Hao LILI ; Zhang ZHANG
Genomics, Proteomics & Bioinformatics 2020;18(2):161-172
Genome reannotation aims for complete and accurate characterization of gene models and thus is of critical significance for in-depth exploration of gene function. Although the availability of massive RNA-seq data provides great opportunities for gene model refinement, few efforts have been made to adopt these precious data in rice genome reannotation. Here we reannotate the rice (Oryza sativa L. ssp. japonica) genome based on integration of large-scale RNA-seq data and release a new annotation system IC4R-2.0. In general, IC4R-2.0 significantly improves the completeness of gene structure, identifies a number of novel genes, and integrates a variety of functional annota-tions. Furthermore, long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) are system-atically characterized in the rice genome. Performance evaluation shows that compared to previous annotation systems, IC4R-2.0 achieves higher integrity and quality, primarily attributable to mas-sive RNA-seq data applied in genome annotation. Consequently, we incorporate the improvedannotations into the Information Commons for Rice (IC4R), a database integrating multiple omics data of rice, and accordingly update IC4R by providing more user-friendly web interfaces and implementing a series of practical online tools. Together, the updated IC4R, which is equipped with the improved annotations, bears great promise for comparative and functional genomic studies in rice and other monocotyledonous species. The IC4R-2.0 annotation system and related resources are freely accessible at http://ic4r.org/.
9. Regulation of angiotensin Ⅱ type 2 receptor in mechanically ventilated lung injury
Shuangyong DONG ; Han HAN ; Yuansheng XU ; Junfeng SONG ; Zhenxi YU ; Jin LI ; Xuyang ZHENG
Chinese Journal of Emergency Medicine 2019;28(12):1511-1516
Objective:
Through the study of angiotensinⅡ- type 2 receptor agonist (AT2R) after pretreatment of mechanical ventilation lung injury (VILI) in rats model, to clarify the role of angiotensin Ⅱ - type 2 receptor agonist (C21) in alleviating VILI inflammation and the damage of immune function and its possible mechanism.
Methods:
In this experiment, the acute lung injury model was established by mechanical spring-volume ventilation in SD rats, and C21 pretreatment was performed to observe the pathological condition of lung tissue in rats with different ventilation duration, and to detect the inflammatory changes of BALF lavage fluid. Flow cytometry was used to detect the CD68+/iNOS+ labeled M1 type AMφ and the CD68+/Arg-1+ labeled M2 type AMφ in alveolar lavage fluid.
Results:
The mechanical VILI rat model was successfully established. The pathological injury score of the mechanical ventilation 4 h model, the wet/dry weight of lung tissue, the number of cells and protein in BALF lavage fluid were increased significantly, the levels of TNF-α and IL - 1 were increased significantly, the levels of IL-4 and IL-10 were decreased significantly, and the level of inflammatory reaction decreased with the increase of ventilation time. The M1/M2 ratio in the 4 h ventilation model group was the highest, which was significantly different from the control group (
10. Prenatal ultrasonic diagnosis and outcome of congenital dacryocystocele
Yixiu ZHANG ; Yuansheng WANG ; Qunni LAMU ; Hua MENG ; Qing DAI ; Zhonghui XU ; Jia LU ; Yunshu OUYANG ; Xining WU ; Yuxin JIANG ; Ji DE ; Yuzhen NIMA
Chinese Journal of Ultrasonography 2018;27(6):520-523
Objective:
To explore the value of prenatal ultrasound in diagnosis of congenital dacryocystocele.
Methods:
The ultrasonographic features of 16 fetuses with congenital dacryocystocele were retrospectively reviewed and the outcome of pregnancy were followed up.
Results:
The median gestational week detected with prenatal ultrasound was 30.29 weeks, the mean diameter was (8.96±1.96)mm. Congenital dacryocystoceles were unilateral in 12 fetuses and bilateral in 4 fetuses, 10 were female and 6 were male. The typical ultrasonic feature was anechoic cystic mass with clear boundary in relation to the medial and inferior aspects of the fetal orbit. The dacryocystocele resolved spontaneously prenatally in 5 fetuses, resolved spontaneously after delivery in 10 fetuses. One fetus died in caesarean section due to complete placenta previa.
Conclusions
Congenital dacryocystitis has its characteristic ultrasonographic features, and most cases can disappear naturally in prenatal or early newborns.

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