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.Teriparatide treatment for osteoporosis:a rapid health technology assessment
Peng LYU ; Jingli ZHANG ; Yuansheng WAN ; Yifei HUANG ; Ruxu YOU
Chinese Journal of Pharmacoepidemiology 2024;33(1):75-84
Objective To evaluate the efficacy,safety and economy of teriparatide in the treatment of osteoporosis.Methods PubMed,Embase,Cochrane Library,Web of Science,CNKI,WanFang Data,VIP databases and websites related to health technology evaluation were systematically searched to collect high-quality clinical evidence and economic evaluation literature of teriparatide in the treatment of osteoporosis from the inception to January 20,2023.Two researchers independently identified studies,extracted data,assessed the quality of included studies,and descriptive analyzed and summarised the results.Results A total of 25 literatures were included,involving 3 HTA reports,15 systematic review/Meta-analyses and 7 economic studies were included.In terms of effectiveness,the evaluation results showed that teriparatide could improve bone mineral density in patients with osteoporosis,reduce the incidence of vertebral/non-vertebral fractures in primary and secondary osteoporosis and prevent the fractures in postmenopausal osteoporosis compared to bisphosphonates and placebos.In terms of safety,teriparatide was proven to be safe with no elevated risk of adverse drug reactions.In terms of economic cost,teriparatide has a higher cost and economic disadvantage compared with bisphosphonates,however,for people with severe postmenopausal osteoporosis and high risk of fracture,teriparatide can be considered as a potential cost-effect treatment option.Conclusion Teriparatide is effective and safe in the treatment of osteoporosis,but it is not cost-effective advantages compared with the existing other anti-osteoporosis medications.
4.Prognostic significance and biological implications of SM‑like genes in mantle cell lymphoma
Xue HE ; Changjian YAN ; Yaru YANG ; Weijia WANG ; Xiaoni LIU ; Chaoling WU ; Zimu ZHOU ; Xin HUANG ; Wei FU ; Jing HU ; Ping YANG ; Jing WANG ; Mingxia ZHU ; Yan LIU ; Wei ZHANG ; Shaoxiang LI ; Gehong DONG ; Xiaoliang YUAN ; Yuansheng LIN ; Hongmei JING ; Weilong ZHANG
Blood Research 2024;59():33-
Background:
SM-like (LSM) genes a family of RNA-binding proteins, are involved in mRNA regulation and can function as oncogenes by altering mRNA stability. However, their roles in B-cell progression and tumorigenesis remain poorly understood.
Methods:
We analyzed gene expression profiles and overall survival data of 123 patients with mantle cell lymphoma (MCL). The LSM index was developed to assess its potential as a prognostic marker of MCL survival.
Results:
Five of the eight LSM genes were identified as potential prognostic markers for survival in MCL, with particular emphasis on the LSM.index. The expression levels of these LSM genes demonstrated their potential utility as classifiers of MCL. The LSM.index-high group exhibited both poorer survival rates and lower RNA levels than did the overall transcript profile. Notably, LSM1 and LSM8 were overexpressed in the LSM.index-high group, with LSM1 showing 2.5-fold increase (p < 0.001) and LSM8 depicting 1.8-fold increase (p < 0.01) than those in the LSM.index-low group.Furthermore, elevated LSM gene expression was associated with increased cell division and RNA splicing pathway activity.
Conclusions
The LSM.index demonstrates potential as a prognostic marker for survival in patients with MCL. Elevated expression of LSM genes, particularly LSM1 and LSM8, may be linked to poor survival outcomes through their involvement in cell division and RNA splicing pathways. These findings suggest that LSM genes may contribute to the aggressive behavior of MCL and represent potential targets for therapeutic interventions.
5.Prognostic significance and biological implications of SM‑like genes in mantle cell lymphoma
Xue HE ; Changjian YAN ; Yaru YANG ; Weijia WANG ; Xiaoni LIU ; Chaoling WU ; Zimu ZHOU ; Xin HUANG ; Wei FU ; Jing HU ; Ping YANG ; Jing WANG ; Mingxia ZHU ; Yan LIU ; Wei ZHANG ; Shaoxiang LI ; Gehong DONG ; Xiaoliang YUAN ; Yuansheng LIN ; Hongmei JING ; Weilong ZHANG
Blood Research 2024;59():33-
Background:
SM-like (LSM) genes a family of RNA-binding proteins, are involved in mRNA regulation and can function as oncogenes by altering mRNA stability. However, their roles in B-cell progression and tumorigenesis remain poorly understood.
Methods:
We analyzed gene expression profiles and overall survival data of 123 patients with mantle cell lymphoma (MCL). The LSM index was developed to assess its potential as a prognostic marker of MCL survival.
Results:
Five of the eight LSM genes were identified as potential prognostic markers for survival in MCL, with particular emphasis on the LSM.index. The expression levels of these LSM genes demonstrated their potential utility as classifiers of MCL. The LSM.index-high group exhibited both poorer survival rates and lower RNA levels than did the overall transcript profile. Notably, LSM1 and LSM8 were overexpressed in the LSM.index-high group, with LSM1 showing 2.5-fold increase (p < 0.001) and LSM8 depicting 1.8-fold increase (p < 0.01) than those in the LSM.index-low group.Furthermore, elevated LSM gene expression was associated with increased cell division and RNA splicing pathway activity.
Conclusions
The LSM.index demonstrates potential as a prognostic marker for survival in patients with MCL. Elevated expression of LSM genes, particularly LSM1 and LSM8, may be linked to poor survival outcomes through their involvement in cell division and RNA splicing pathways. These findings suggest that LSM genes may contribute to the aggressive behavior of MCL and represent potential targets for therapeutic interventions.
6.Prognostic significance and biological implications of SM‑like genes in mantle cell lymphoma
Xue HE ; Changjian YAN ; Yaru YANG ; Weijia WANG ; Xiaoni LIU ; Chaoling WU ; Zimu ZHOU ; Xin HUANG ; Wei FU ; Jing HU ; Ping YANG ; Jing WANG ; Mingxia ZHU ; Yan LIU ; Wei ZHANG ; Shaoxiang LI ; Gehong DONG ; Xiaoliang YUAN ; Yuansheng LIN ; Hongmei JING ; Weilong ZHANG
Blood Research 2024;59():33-
Background:
SM-like (LSM) genes a family of RNA-binding proteins, are involved in mRNA regulation and can function as oncogenes by altering mRNA stability. However, their roles in B-cell progression and tumorigenesis remain poorly understood.
Methods:
We analyzed gene expression profiles and overall survival data of 123 patients with mantle cell lymphoma (MCL). The LSM index was developed to assess its potential as a prognostic marker of MCL survival.
Results:
Five of the eight LSM genes were identified as potential prognostic markers for survival in MCL, with particular emphasis on the LSM.index. The expression levels of these LSM genes demonstrated their potential utility as classifiers of MCL. The LSM.index-high group exhibited both poorer survival rates and lower RNA levels than did the overall transcript profile. Notably, LSM1 and LSM8 were overexpressed in the LSM.index-high group, with LSM1 showing 2.5-fold increase (p < 0.001) and LSM8 depicting 1.8-fold increase (p < 0.01) than those in the LSM.index-low group.Furthermore, elevated LSM gene expression was associated with increased cell division and RNA splicing pathway activity.
Conclusions
The LSM.index demonstrates potential as a prognostic marker for survival in patients with MCL. Elevated expression of LSM genes, particularly LSM1 and LSM8, may be linked to poor survival outcomes through their involvement in cell division and RNA splicing pathways. These findings suggest that LSM genes may contribute to the aggressive behavior of MCL and represent potential targets for therapeutic interventions.
7.Changes of complement fragment C3a and its receptor in lung injury induced by silica nanoparticles
Sifan ZHOU ; Zhoujian WANG ; Lei GAO ; Yuansheng DUAN ; Muyue CHEN ; Jiaxiang ZHANG ; Qixing ZHU
Journal of Environmental and Occupational Medicine 2023;40(8):877-883
Background Silica nanoparticles (SiNPs) enter the human body through respiratory tract, digestive tract, and skin, causing body damage. Lung is one of the main damaged organs. Objective To observe the expressions of complement activated fragment C3a and its receptor C3aR in the lungs of mice exposed to SiNPs through respiratory tract, and to explore the involvement of C3a/C3aR in lung injury induced by SiNPs exposure. Methods The ultrastructure of SiNPs (particle size 5-20 nm) was determined under a transmission electron microscope, and the hydrodynamic diameter and surface Zeta potential of SiNPs were determined using a nanoparticle size analyzer. A total of 88 SPF C57BL/6J mice were randomly divided into five groups: a blank control group without any treatment (14 mice), a vehicle control group treated with 50 μL stroke-physiological saline solution by intratracheal instillation (14 mice), and three SiNPs exposure groups (low-dose group, medium-dose group, and high-dose group with 20 mice in each group, who were given 50 μL SiNPs suspension of 7, 21, and 35 mg·kg−1 respectively and exposed once every 3 days for 5 times). The mice were anesthetized on day 1 (1-day model group) and day 15 (15-day model group) after exposure, then sacrificed after extraction of bronchoalveolar lavage fluid (BALF), and lung tissues were retained. The morphological changes of lung tissues were observed by HE staining, the expression level of C3a in BALF was detected by enzyme-linked immunosorbent assay, the deposition of C3a and C3aR in lung tissues were observed by immunohistochemistry, the protein expression level of C3aR was determined by Western blotting, and the localization and semi-quantitative detection of C3a and C3aR in lung tissues was observed by immunofluorescence. Results SiNPs agglomerated in stroke-physiological saline solution. The average hydrodynamic diameter was (185.60±7.39) nm and the absolute value of Zeta potential was (43.33±0.76) mV. The condition of mice in the 1-day model group and the 15-day model group was good, while 2 mice died in the medium-dose group of the 1-day model group due to misoperation. The autopsy results of the two mice showed congestion of the lung tissue, emphysema, and no imperfection of trachea integrity. No death was observed in other dose groups. The HE staining results showed pathological damage to the mouse lung, including alveolar wall thickening and inflammatory cell infiltration after SiNPs exposure. The pathological damage became more serious with the increase of dose. Regarding pathological changes, the 15-day model group was slightly relieved compared with the 1-day model group, but there were still pathological changes. The enzyme-linked immunosorbent assay results showed that there was no difference in the expression level of C3a between the blank control group and the vehicle control group (P>0.05), the expression levels of C3a in the medium-dose group and the high-dose group were significantly higher than that in the vehicle control group (P<0.05). The immunohistochemistry results showed that C3a deposition was consistent with the enzyme-linked immunosorbent assay results. The Western blotting and the immunohistochemistry results showed that C3aR expression was low in the blank control group and the vehicle control group, while the expression in each dose group tended to increase with the increase of dose. The immunofluorescence results showed that the fluorescence signals of C3a and C3aR were weak in the blank control group and the vehicle control group in the 1-day model group and the 15-day model group, while the fluorescence signals in the lung tissues of mice in the SiNPs exposure groups tended to increase with the increase of dose. Conclusion The increased expressions of C3a and C3aR in complement activation may be related to lung injury induced by intratracheal instillation of SiNPs, suggesting that C3a/C3aR may be involved in lung injury induced by SiNPs exposure.
8.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.
9.Experimental study on optimization of mAs in digital mammography
Yuansheng ZHANG ; Xiaocheng WANG ; Ziquan GUO ; Jianming WANG
Chinese Journal of Radiological Medicine and Protection 2022;42(2):144-149
Objective:To evaluate the effect of reducing mAs on image quality when different target/filter combinations are used in digital mammography.Methods:In different target/filter combinations, based on the mAs of automatic exposure control(AEC), the reduction of mAs by 10%, 20%, 30%, 40% and 50% respectively were used to expose the phantom with 4.4 cm thickness. The contrast to noise ratio(CNR), signal to noise ratio(SNR), figure of merit(FOM)and average glandular dose(AGD) of the resulting image were calculated. While the image quality was ensured, the optimal mAs and the corresponding AGD under the two target/filter combination were conducted by calculating the FOM. The image features of three tissues of phantom were subjectively evaluated, and the relationship between the calculated AGD and displayed AGD was compared. The mAs that meets the image quality requirements is corresponded with the density exposure gear, and the average values of the mAs for two method were calculated and compared.Results:In two targets/filters combinations, the displayed AGD was less than the calculated AGD, and Mo/Mo was underestimated by 22.5% to 23.7%. The calculated and displayed AGD values were statistically different ( F=4 982.86, 5 555.48, P<0.05). W/Rh was underestimated by 13.1% to 14.2%. The calculated and displayed AGD values were statistically different ( F=18 859.09, 15 973.55, P<0.05). In the Mo/Mo combination, when the mAs was reduced by 20%, the FOM could be increased by 9.6% for the maximum value, and the calculated AGD was decreased by 18.8%. In the W/Rh combination, when the mAs was reduced by 10%, the FOM was increased by 5.1% for the maximum value, and the calculated AGD was decreased by 11.9%. While the image quality was ensured, the mAs was reduced by 30% for evaluating simulated fibers, and by 20% for evaluating simulated specks in the two targets/filter combinations. For evaluating simulated masses, Mo/Mo combination reduced the mAs by 40%, and the W/Rh combination reduced the mAs by 30%. And the image quality scores in above were not significantly different from those in AEC method ( P>0.05). The density exposure gear(-1 to -3) could correspond with the reduced mAs(10%-30%). Conclusions:Under different target/filter combination, the tube mAs could be reduced when the image quality was ensured.
10.Effects of HDAC6 regulation of HSC70 acetylation level on bone loss in osteoporosis after oophorectomy
Hui LI ; Jieyun LEI ; Li ZHANG ; Shuangxue LI ; Yuansheng LEI ; Feng LI
Chinese Journal of Endocrine Surgery 2022;16(2):190-195
Objective:To investigate the effects of histone deacetylase 6 (histone deacetylase 6, HDAC6) on oopherectomy (OOX) induced osteoporosis (OP) bone loss by binding to the promoter region of heat-shock protein 70 (HSC70) and regulating it’s acetylation.Methods:OP mouse model was established by using OOX methods. Then the mice were divided into sham operation group, OOX group, OOX+shHDAC6 group, OOX+shNC group and OOX+shHDAC6+shHSC70 group. The micro-CT system and Western blot experiment were used to detect the bone microscopic parameters of the mouse right femur and the protein expression levels of osteoblast-specific transcription factors. In vitro experiments, Westwen blot, alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining were used to determine the effects of HDAC6 and HSC70 on the osteogenic differentiation of MC3T3-E1 cells. QRT-PCR was used to detect the expression levels of HDAC6 and HSC70 in tissue or cells. The relationship between HDAC6 and HSC70 was analyzed by ChIP experiment.Results:Compared with sham group, the expression of bone mineral density (BMD) , trabecular bone number (Tb. N) , trabecular thickness (Tb.th) and bone volume fraction (BV/TV) in the right femur of OOX group mice were decreased, the expression of TB. Sp was increased, protein expression of OSX and RUNX2 was increased. At the same time, compared with sham group (1±0.11) , the expression of HDAC6 was increased in OOX group (2.33±0.19) ( t=10.56, P<0.001) . Compared with pcDNA3.1-NC group, the protein level of Osterix (OSX) and runt-related transcription factor 2 (RUNX2) , ALP activity and mineralized area in pcDNA3.1-HDAC6 group were decreased (all P<0.05) . ChIP analysis showed that compared with the pcDNA3.1-NC group (5.26±0.47) , the acetylation level of the HSC70 promoter region in the pcDNA3.1-HDAC6 group (2.37±0.21) was significantly reduced ( t=9.72, P<0.001) . Compared with pcDNA3.1-HDAC6 group, the expression of OSX and RUNX2, ALP activity and mineralization were increased in pcDNA3.1-HDAC6+ pcDNA3.1-HSC70 group (all P<0.05) . Compared with OOX+shHDAC6 group, the expression of OSX and RUNX2 protein, BMD, Tb.N, Tb.th and BV/TV were decreased but the expression of Tb. Sp was increased in OOX+ shHDAC6+ shHSC70 group. Conclusions:HDAC6 regulates the acetylation level of HSC70 and then affects OOX-induced OP bone loss. Inhibition of HDAC6 can significantly improve OP bone loss.

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