1.Analysis of Quality Uniformity of Hengzhi Kechuan Capsules Based on HPLC-DAD-CAD
Qian MA ; An LIU ; Qingxia XU ; Cong GUO ; Jun ZHANG ; Maoqing WANG ; Xiaodi KOU ; Yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):168-174
ObjectiveTo establish the fingerprints of 15 batches of Hengzhi Kechuan capsules, to quantitatively analyze 10 index components, and to evaluate the quality uniformity of samples from different batches. MethodsThe fingerprints and quantitative analysis of Hengzhi Kechuan capsules were established by a combination method of high performance liquid chromatography coupled with diode array detector and charged aerosol detector(HPLC-DAD-CAD), adenosine, guanosine, vanillic acid, safflomin A, agarotetrol, naringin, hesperidin, militarine, ginsenoside Rb1, and glycyrrhizic acid were selected as quality attribute indexes. A total of 15 batches of Hengzhi Kechuan capsules from 2022 to 2024(3 boxes per batch) were qualitatively and quantitatively analyzed, and the quality uniformity level of the manufacturers was characterized by parameters of intra-batch consistency(PA) and inter-batch consistency(PB). The homogeneity and difference of quality attribute indexes of samples from different years were analyzed by heatmap clustering analysis. ResultsHPLC fingerprints and quantitative method of Hengzhi Kechuan capsules were established, and the methods could be used for qualitative and quantitative analysis of this preparation, which was found to be stable and reliable by method validation. The similarity of fingerprints of 15 batches of samples was 0.887-0.975, a total of 13 common peaks were calibrated, and 10 common peaks were designated, all of which were quality attribute index components. The results of quantitative analysis showed that the contents of the above 10 ingredients in the samples were 0.038-0.078, 0.115-0.251, 0.007-0.018, 0.291-0.673, 0.122-0.257, 0.887-1.905, 1.841-3.364, 1.412-2.450, 2.207-3.112, 0.650-1.161, respectively. And the contents of ginsenoside Rb1 and glycyrrhizic acid met the limit requirements in the 2020 edition of Chinese Pharmacopoeia. For the samples from 15 batches, the PA values of the 10 index components were all <10%, indicating good intra-batch homogeneity, and the PB values ranged from 33.86% to 92.97%, suggesting that the inter-batch homogeneity was poor. Heatmap clustering analysis showed that the samples from different years were clustered into separate categories, and adenosine, guanosine, safflomin A, naringin, hesperidin and agarotetrol were the main differential components. ConclusionThe intra-annual quality uniformity of Hengzhi Kechuan capsules is good and the inter-annual quality uniformity is insufficient, which may be related to the quality difference of Pinellinae Rhizoma Praeparatum, Carthami Flos, Citri Sarcodactylis Fructus, Citri Reticulatae Pericarpium, Aquilariae Lignum Resinatum, Citri Fructus, etc. In this study, the fingerprint and multi-indicator determination method of Hengzhi Kechuan capsules was established, which can be used for more accurate and efficient quality control and standardization enhancement.
2.Construction and identification of recombinant fowl adenovirus 4 expressing Cap protein of goose astrovirus virus genotype 2
Xingyu LI ; Yan LI ; Panpan YANG ; Junjie LIU ; Mengjia XIANG ; Yutao ZHU ; Luyao QIU ; Qilong QIAO ; Boshun ZHANG ; Dexin BU ; Chenghao HAN ; Chunmei YU ; Yanfang CONG ; Zeng WANG ; Jianli LI ; Baiyu WANG ; Jun ZHAO
Chinese Journal of Veterinary Science 2025;45(3):443-448,513
To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.
3.Protective effects of Shuangyi Qushi Tongluo Capsules on dexamethasone-induced osteoporosis in mice
Yi LI ; Jian-bin HE ; Jia-xiu XIE ; Quan-mou LUO ; Dong-mei LI ; Jun-hui HE ; Dong-mei WEI ; Chao WEI ; Hong-cong QIU ; Gui-ning WEI ; Bo WANG
Chinese Traditional Patent Medicine 2025;47(6):1834-1842
AIM To investigate the protective effects of Shuangyi Qushi Tongluo Capsules(Shuangyi Capsules)on Dexamethasone(Dex)induced osteoporosis in mice.METHODS The C57BL/6J mice were randomly divided into the control group,the model group,the Xianling Gubao Capsules group(1.5 g/kg),and the low-dose,moderate-dose,and high-dose Shuangyi Capsules groups(0.6,1.2,and 2.4 g/kg).The mouse model of osteoporosis was induced by 8-week intraperitoneal injection of Dex sodium phosphate injection(5 mg/kg).The mice had their femur osteogenesis observed with hematoxylin and eosin(HE)staining and tartrate-resistant acid phosphatase(TRAP)staining;their serum alkaline phosphatase(ALP)and osteocalcin(BGP)activities detected by ELISA;their femoral mRNA expressions of Col-Ⅰ,OCN,and OPN detected by RT-qPCR;and their femoral protein expressions of OPG and RANKL detected by Western blot.Upon the MC3T3-E1 cells exposed to Dex and Shuangyi Capsules,their viability was evaluated by CCK-8 assay;their mineralization determined by alkaline phosphatase staining and alizarin red staining(ARS);and their intracellular ROS level detected using DCFH-DA probe.RESULTS Compared with the model group,Shuangyi Capsules groups demonstrated improved fracture of femoral trabeculae and reduced number of osteoclasts;increased serum ALP and BGP activities(P<0.05,P<0.01);increased femoral expressions of Col-Ⅰ mRNA and OPG protein(P<0.05,P<0.01);and decreased RANKL protein expression(P<0.05).Compared with the MC3T3-E1 cells stimulated by Dex,those underwent further treatment of Shuangyi Capsules demonstrated increased cell viability and ALP activity(P<0.05,P<0.01);increased mineralization and calcium nodule formation;increased expressions of Col-Ⅰ,OCN,OPN mRNA and OPG protein(P<0.05,P<0.01);decreased RANKL protein expression(P<0.05,P<0.01);and reduced ROS levels.CONCLUSION Shuangyi Capsules ameliorate Dex-induced osteoporosis in mice by suppressing osteoclast overactivation,enhancing osteoblast activity,and stimulating bone formation through modulation of Col-Ⅰ,OCN,OPN mRNA and OPG/RANKL protein levels.
4.Connotation and Prevention Strategies of Traditional Chinese Medicine for Panvascular Diseases
Jie WANG ; Jun LI ; Yan DONG ; Cong CHEN ; Yongmei LIU ; Chao LIU ; Lanchun LIU ; Xuan SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):1-14
Panvascular disease, with vascular diseases as the common pathological feature, is mainly manifested as atherosclerosis. Panvascular disease mainly affects the important organs of the heart, brain, kidney, and limbs. It is one of the leading causes of death for Chinese residents at present. Previously, due to the narrow branches of disciplines, too much attention was paid to local lesions, resulting in the neglect of panvascular disease as a systemic one. The fact that panvascular disease has overall pathology and comprehensive and individualized treatment strategies, makes the disease highly compatible with the principles of holism concept and syndrome differentiation and treatment in traditional Chinese medicine (TCM). It is believed that blood stasis is the core pathogenesis of atherosclerosis and is involved in the whole process of atherosclerosis. The theories of ''blood vessel'', ''meridians'', ''visceral manifestation'', and ''organs-meridians'' in TCM are helpful to comprehensively understand the complexity of panvascular diseases. Moreover, those theories can provide systematic treatment strategies. The TCM syndromes of panvascular diseases evolve from ''phlegm, stasis, stagnation, and deficiency''. Panvascular arteriosclerosis is related to the syndrome of ''stasis and phlegm'', and the treatment mainly promotes blood circulation and removes phlegm. There are different specific drugs and mechanisms of action for coronary atherosclerosis, cerebral atherosclerosis, and renal artery atherosclerotic stenosis. Panvascular venous lesions are related to the syndrome of ''deficiency and stasis'' in TCM, and the TCM treatment mainly invigorates Qi and promotes blood circulation, which can inhibit venous thrombosis, improve venous ulcers, and resist venous endothelial damage. Panvascular microcirculatory lesions are inseparable from the ''stagnation and stasis'' in TCM, and the treatment mainly promotes Qi and dredges collaterals, which has a good effect on coronary microvascular lesions, diabetic microvascular lesions, pulmonary microvascular lesions, and pancreatic microvascular lesions. Panvascular lymphatic lesions are related to the syndrome of ''water and stasis'' in TCM. The treatment method focuses on promoting blood circulation and water excretion, which can promote lymphangiogenesis and enhance lymphatic reflux. In addition, the combination of TCM and modern technology, especially the application of artificial intelligence, can improve the efficiency of early identification and personalized treatment, resulting in early screening and comprehensive management of panvascular diseases. Therefore, TCM will play a vital role in the prevention and treatment of panvascular diseases.
5.Connotation and Prevention Strategies of Traditional Chinese Medicine for Panvascular Diseases
Jie WANG ; Jun LI ; Yan DONG ; Cong CHEN ; Yongmei LIU ; Chao LIU ; Lanchun LIU ; Xuan SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):1-14
Panvascular disease, with vascular diseases as the common pathological feature, is mainly manifested as atherosclerosis. Panvascular disease mainly affects the important organs of the heart, brain, kidney, and limbs. It is one of the leading causes of death for Chinese residents at present. Previously, due to the narrow branches of disciplines, too much attention was paid to local lesions, resulting in the neglect of panvascular disease as a systemic one. The fact that panvascular disease has overall pathology and comprehensive and individualized treatment strategies, makes the disease highly compatible with the principles of holism concept and syndrome differentiation and treatment in traditional Chinese medicine (TCM). It is believed that blood stasis is the core pathogenesis of atherosclerosis and is involved in the whole process of atherosclerosis. The theories of ''blood vessel'', ''meridians'', ''visceral manifestation'', and ''organs-meridians'' in TCM are helpful to comprehensively understand the complexity of panvascular diseases. Moreover, those theories can provide systematic treatment strategies. The TCM syndromes of panvascular diseases evolve from ''phlegm, stasis, stagnation, and deficiency''. Panvascular arteriosclerosis is related to the syndrome of ''stasis and phlegm'', and the treatment mainly promotes blood circulation and removes phlegm. There are different specific drugs and mechanisms of action for coronary atherosclerosis, cerebral atherosclerosis, and renal artery atherosclerotic stenosis. Panvascular venous lesions are related to the syndrome of ''deficiency and stasis'' in TCM, and the TCM treatment mainly invigorates Qi and promotes blood circulation, which can inhibit venous thrombosis, improve venous ulcers, and resist venous endothelial damage. Panvascular microcirculatory lesions are inseparable from the ''stagnation and stasis'' in TCM, and the treatment mainly promotes Qi and dredges collaterals, which has a good effect on coronary microvascular lesions, diabetic microvascular lesions, pulmonary microvascular lesions, and pancreatic microvascular lesions. Panvascular lymphatic lesions are related to the syndrome of ''water and stasis'' in TCM. The treatment method focuses on promoting blood circulation and water excretion, which can promote lymphangiogenesis and enhance lymphatic reflux. In addition, the combination of TCM and modern technology, especially the application of artificial intelligence, can improve the efficiency of early identification and personalized treatment, resulting in early screening and comprehensive management of panvascular diseases. Therefore, TCM will play a vital role in the prevention and treatment of panvascular diseases.
6.Innate immune cell LXR-β deficiency exacerbates hepatic injury and fibrosis in murine models of primary sclerosing cholangitis
Xiaohui FANG ; Yang ZHANG ; Junyao WANG ; Yu ZHANG ; Ziliang KE ; Yiken LIN ; Fangyuan CONG ; Feng ZHANG ; Jianhua ZHOU ; Huiting SU ; Shan CAO ; Yulan LIU ; Jun XU
Liver Research 2025;9(3):239-248
Background and aims:Primary sclerosing cholangitis(PSC)is an autoimmune liver disease characterized by complex pathogenesis and limited available therapeutic options.The mechanisms underlying the development and progression of PSCs remain unclear.Liver X receptor beta(LXR-β)is recognized to modulate lipid metabolism and immune response,but its specific involvement in the PSC has not been elucidated.Here,we explored the role and mechanism of LXR-β in PSC induced by 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-collidine(DDC).Methods:CRISPR-Cas9 technology was applied to generate Abcb4(coding MDR2,next named as Mdr2),Nr1h2(coding LXR-β,next named as Lxrβ),and Rag2(coding RAG2)knockout mice.DDC was used to induce PSC.Hematoxylin and eosin and Sirius red staining were used to assess the extent of hepatic injury and fibrosis.Flow cytometry was used to observe immune cell subsets.Results:We observed a declining trend in hepatic Lxrβ in the PSC model.Unexpectedly,Lxrβ knockout failed to modulate DDC-induced PSC pathogenesis.Concomitantly,assessment of the influence of Rag2 deficiency on PSC progression revealed the absence of aggravated or alleviated hepatic injury or fibrosis in the Rag2-/-DDC mice.However,Lxrβ depletion intensified DDC-induced PSC in the Rag2-/-mice,with more abundant infiltrative inflammatory cells and more severe liver fibrosis.Compared with Rag2-/-DDC mice,Lxrβ-/-Rag2-/-DDC mice had higher serum ALT and AST levels and mRNA expression of proinflammatory and profibrotic genes.Flow cytometry showed that LXR-β deficiency resulted in a diminished population of hepatic innate immune cells.Conclusion:This study indicated innate immune cell LXR-β deficiency can exacerbate hepatic injury and fibrosis in murine models of PSC suggesting that LXR-β may regulate the function of innate immunity in the fibrotic advancement of PSC.
7.Effect of preoperative anxiety on laparoscopic hysterectomy under general anesthesia in the elderly
Jing-hui QIU ; Hui-rong DAI ; Lin JI ; Cong-hu YUAN ; Tao WANG ; Ti-jun DAI
Journal of Regional Anatomy and Operative Surgery 2025;34(3):228-231
Objective To investigate the effects of preoperative anxiety on the intraoperative body temperature,recessive blood loss,and wound healing in elderly patients undergoing laparoscopic hysterectomy under general anesthesia.Methods A total of 118 elderly patients who underwent laparoscopic hysterectomy in Yancheng Third People's Hospital from January 2021 to December 2023 were selected as the research subjects,and they were divided into the normal group(69 cases with normal emotions)and the anxiety group(49 cases with anxiety emotions)according to the self-rating anxiety scale(SAS)score 1 day before operation.The core temperature at different time points and the incidence of intraoperative hypothermia were compared between the two groups.The intraoperative bleeding volume,incision drainage bleeding volume,recessive blood loss,incidence of adverse reactions,and wound healing were compared between the two groups.Results The core temperature 60 minutes after operation and at the end of operation in the anxiety group were significantly lower than those in the normal group(P<0.05).The incidence of intraoperative hypothermia in the anxiety group was significantly higher than that in the normal group(P<0.05).There was no statistically significant difference in the intraoperative bleeding volume or incision drainage bleeding volume between the two groups(P>0.05),while the recessive blood loss in the anxiety group was significantly higher than that in the normal group(P<0.05).The incidence of shivering in the anxiety group was significantly higher than that in the normal group(P<0.05);the incidence of poor wound healing in the anxiety group was significantly higher than that in the normal group(P<0.05).Conclusion For elderly patients who undergo laparoscopic hysterectomy for more than 60 minutes,the body temperature will significantly decrease,the incidence of intraoperative hypothermia and shivering is high,the recessive blood loss will significantly increase,which is prone to poor wound healing and is not conducive to the rapid recovery of patients after operation,if combined with anxiety before surgery.
8.Protective effects of Shuangyi Qushi Tongluo Capsules on dexamethasone-induced osteoporosis in mice
Yi LI ; Jian-bin HE ; Jia-xiu XIE ; Quan-mou LUO ; Dong-mei LI ; Jun-hui HE ; Dong-mei WEI ; Chao WEI ; Hong-cong QIU ; Gui-ning WEI ; Bo WANG
Chinese Traditional Patent Medicine 2025;47(6):1834-1842
AIM To investigate the protective effects of Shuangyi Qushi Tongluo Capsules(Shuangyi Capsules)on Dexamethasone(Dex)induced osteoporosis in mice.METHODS The C57BL/6J mice were randomly divided into the control group,the model group,the Xianling Gubao Capsules group(1.5 g/kg),and the low-dose,moderate-dose,and high-dose Shuangyi Capsules groups(0.6,1.2,and 2.4 g/kg).The mouse model of osteoporosis was induced by 8-week intraperitoneal injection of Dex sodium phosphate injection(5 mg/kg).The mice had their femur osteogenesis observed with hematoxylin and eosin(HE)staining and tartrate-resistant acid phosphatase(TRAP)staining;their serum alkaline phosphatase(ALP)and osteocalcin(BGP)activities detected by ELISA;their femoral mRNA expressions of Col-Ⅰ,OCN,and OPN detected by RT-qPCR;and their femoral protein expressions of OPG and RANKL detected by Western blot.Upon the MC3T3-E1 cells exposed to Dex and Shuangyi Capsules,their viability was evaluated by CCK-8 assay;their mineralization determined by alkaline phosphatase staining and alizarin red staining(ARS);and their intracellular ROS level detected using DCFH-DA probe.RESULTS Compared with the model group,Shuangyi Capsules groups demonstrated improved fracture of femoral trabeculae and reduced number of osteoclasts;increased serum ALP and BGP activities(P<0.05,P<0.01);increased femoral expressions of Col-Ⅰ mRNA and OPG protein(P<0.05,P<0.01);and decreased RANKL protein expression(P<0.05).Compared with the MC3T3-E1 cells stimulated by Dex,those underwent further treatment of Shuangyi Capsules demonstrated increased cell viability and ALP activity(P<0.05,P<0.01);increased mineralization and calcium nodule formation;increased expressions of Col-Ⅰ,OCN,OPN mRNA and OPG protein(P<0.05,P<0.01);decreased RANKL protein expression(P<0.05,P<0.01);and reduced ROS levels.CONCLUSION Shuangyi Capsules ameliorate Dex-induced osteoporosis in mice by suppressing osteoclast overactivation,enhancing osteoblast activity,and stimulating bone formation through modulation of Col-Ⅰ,OCN,OPN mRNA and OPG/RANKL protein levels.
9.Analysis of Quality Difference Factors of Perillae Caulis Based on Chemometrics Combined with TOPSIS Model
Maoqing WANG ; Sha CHEN ; Qian MA ; Jun ZHANG ; Qingxia XU ; Cong GUO ; Rui SHEN ; Yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):168-175
ObjectiveTo explore quality difference factors of Perillae Caulis based on the contents of multiple chemical components and comprehensively evaluate the quality. MethodsA total of 32 batches of Perillae Caulis samples were collected from 12 producing areas such as Hebei, Anhui and Guangdong, and their diameter range, epidermis color and producing areas were recorded. Total flavonoids, total phenols, volatile oils, 5 active components and 84 volatile components in 32 batches of samples were quantitatively or semi-quantitatively determined by colorimetry, ultra performance liquid chromatography-photodiode array detector(UPLC-PDA) and gas chromatography-mass spectrometry(GC-MS). Then the differences between the contents of these components were analyzed by principal component analysis(PCA) and non-parametric test. According to the weights of the index components determined by PCA model, entropy weight-technique for order preference by similarity to ideal solution(TOPSIS) model was constructed to evaluate the quality of Perillae Caulis with different characters and origins. ResultsThere were significant differences in the composition of Perillae Caulis with different diameters, epidermis colors and producing areas, and 9 differential components were screened out, including 6 index constituents(total flavonoids, total phenols, caffeic acid, scutellarin, rosmarinic acid and luteolin) and 3 volatile components(caryophyllene oxide, (-)-humulene epoxide Ⅱ, 14-hydroxycaryophyllene), of which 6 index constituents were higher in samples with small diameter, purple-brown epidermis and southern origin, while the contents of 3 volatile components were higher in samples with large diameter, dark-brown epidermis and northern origin. A significant difference was shown in the model scores of different diameters, epidermis colors and origins(P<0.05), and the scores of Perillae Caulis with small diameter and purple-brown epidermis from southern area, especially Guangdong, had a high score. ConclusionThere are significant differences in the composition and content of chemical constituents between different diameters, epidermal colors and production areas of Perillae Caulis, samples showing small diameter, owing purple-brown epidermis, and originating from Guangdong were of higher-quality due to their higher content of 8 key indices.
10.Analysis of Quality Difference Factors of Perillae Caulis Based on Chemometrics Combined with TOPSIS Model
Maoqing WANG ; Sha CHEN ; Qian MA ; Jun ZHANG ; Qingxia XU ; Cong GUO ; Rui SHEN ; Yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):168-175
ObjectiveTo explore quality difference factors of Perillae Caulis based on the contents of multiple chemical components and comprehensively evaluate the quality. MethodsA total of 32 batches of Perillae Caulis samples were collected from 12 producing areas such as Hebei, Anhui and Guangdong, and their diameter range, epidermis color and producing areas were recorded. Total flavonoids, total phenols, volatile oils, 5 active components and 84 volatile components in 32 batches of samples were quantitatively or semi-quantitatively determined by colorimetry, ultra performance liquid chromatography-photodiode array detector(UPLC-PDA) and gas chromatography-mass spectrometry(GC-MS). Then the differences between the contents of these components were analyzed by principal component analysis(PCA) and non-parametric test. According to the weights of the index components determined by PCA model, entropy weight-technique for order preference by similarity to ideal solution(TOPSIS) model was constructed to evaluate the quality of Perillae Caulis with different characters and origins. ResultsThere were significant differences in the composition of Perillae Caulis with different diameters, epidermis colors and producing areas, and 9 differential components were screened out, including 6 index constituents(total flavonoids, total phenols, caffeic acid, scutellarin, rosmarinic acid and luteolin) and 3 volatile components(caryophyllene oxide, (-)-humulene epoxide Ⅱ, 14-hydroxycaryophyllene), of which 6 index constituents were higher in samples with small diameter, purple-brown epidermis and southern origin, while the contents of 3 volatile components were higher in samples with large diameter, dark-brown epidermis and northern origin. A significant difference was shown in the model scores of different diameters, epidermis colors and origins(P<0.05), and the scores of Perillae Caulis with small diameter and purple-brown epidermis from southern area, especially Guangdong, had a high score. ConclusionThere are significant differences in the composition and content of chemical constituents between different diameters, epidermal colors and production areas of Perillae Caulis, samples showing small diameter, owing purple-brown epidermis, and originating from Guangdong were of higher-quality due to their higher content of 8 key indices.

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