1.Determination and Comparison of 26 Components in Different Parts of Two Base Plants of Shiliang Tea Based on UHPLC-MS/MS
Juhua MAO ; Junjie PAN ; Zhangjin CHEN ; Chaqing WU ; Weiying WANG ; Kejun CHENG
Chinese Journal of Modern Applied Pharmacy 2024;41(4):489-495
OBJECTIVE
To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method for determination and comparison of 26 components in different parts of two base plants of Shiliang tea(Chimonanthus salicifolius S.Y.Hu and Chimonanthus zhejiangensis M.C.Liu), and screen quality markers of different parts.
METHODS
The UHPLC method was performed on an Agilent RRHD Eclipse Plus C18 (2.1 mm×50 mm, 1.8 μm) column with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.3 mL·min−1, the column temperature was 35 ℃, and the injection volume was 0.5 μL; the multiple reaction monitoring mode was employed for the quantification of 26 components with electrospray ionization(ESI) source polarity in negative and positive mode.
RESULTS
Good linear relationship(r >0.999) were observed in the test ranges for 26 compounds, and the average recovery was 88.5%−111.7% with RSD was 3.4%−9.8%. There was no significant difference between the two base plants of Shiliang tea, and all of these samples were divided into two categories by hierarchical cluster analysis. The main components in leaves was flavonoids, among them, the content of kaempferol 3-O-rutinoside was the highest, reaching 12.902 mg.g−1; the main components in stems and roots was coumarins, and the content of alkaloids in roots was higher, relatively; 7 quality markers of difference were screened by OPLS-DA, which were kaempferol 3-O-rutinoside, chimonanthine, rutin, fraxetin, calycanthoside, scopolin, neochlorogenic acid.
CONCLUSION
These study elucidates the differences of chemical components in the different parts of two base plants of Shiliang tea, which providing basis for the research of pharmacodynamic substances and references for the comprehensive utilization of Chimonanthus salicifolius S.Y. Hu and Chimonanthus zhejiangensis M.C.Liu resources.
2.Huangjing Jiannao Granules Improve Learning and Memory Abilities and Cerebral Blood Flow in Rat Model of Vascular Cognitive Impairment via PI3K/Akt Signaling Pathway
Rui YANG ; Yumu TIAN ; Yujing JIN ; Jianwen ZHAIWU ; Tong ZHANG ; Zehua ZHAO ; Shijing HUANG ; Juhua PAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):52-60
ObjectiveTo study the effects of Huangjing Jiannao granules on learning and memory abilities and cerebral blood flow in the rat model of vascular cognitive impairment (VCI) and to explore the mechanism of Huangjing Jiannao granules in the treatment of VCI. MethodSeventy-two SPF-grade male SD rats were randomly selected, with 12 rats as the sham operation group. The remaining rats were subjected to bilateral carotid artery ligation (2-VO) for the modeling of VCI. According to the randomized block design, the successfully modeled rats were grouped as follows: model, donepezil hydrochloride (0.50 mg·kg-1), and low-, medium-, and high-dose (2.36, 4.72, 9.44 g·kg-1, respectively) Huangjing Jiannao granules. After 6 weeks of treatment, Morris water maze test and new object recognition test were conducted to evaluate the learning and memory abilities of the rats. After continuous gavage for 8 weeks, the cerebral blood flow was recorded by a laser microcirculation blood flow imager, and the survival and injury of hippocampal neurons were observed by Nissl staining. The expression of neuronal nuclear antigen (NeuN) in the hippocampus was detected by immunohistochemistry (IHC). The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the serum were determined by enzyme-linked immunosorbent assay. The protein levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), nuclear factor-κB p65 (NF-κB p65), and nuclear factor-κB inhibitor α (IκBα) in the hippocampus were determined by Western blot. ResultCompared with the sham operation group, the model group showed weakened learning and memory abilities (P<0.01), reduced blood flow in the whole brain, forebrain, and hindbrain (P<0.01), damaged neurons and reduced survived neurons in the hippocampal CA1 region (P<0.01), down-regulated expression of NeuN (P<0.01), elevated levels of IL-1β and TNF-α in the serum (P<0.01), up-regulated protein levels of PI3K, p-Akt, and NF-κB p65 in the hippocampal tissue, and down-regulated protein level of IκBα (P<0.01). Compared with the model group, medium- and high-dose Huangjing Jiannao granules improved the learning and memory abilities (P<0.05,P<0.01). High-dose Huangjing Jiannao granules increased the blood flow in the whole brain, forebrain, and hindbrain (P<0.05,P<0.01), and medium-dose Huangjing Jiannao granules increased the blood flow in the whole brain (P<0.05). All the doses of Huangjing Jiannao granules increased the number of survived neurons (P<0.05,P<0.01) and up-regulated the protein level of NeuN (P<0.05,P<0.01). Medium and high-dose Huangjing Jiannao granules lowered the level of TNF-α (P<0.05,P<0.01), down-regulated the protein levels of PI3K, p-Akt, and NF-κB p65 (P<0.05,P<0.01), and up-regulated the protein level of IκBα (P<0.01). ConclusionHuangjing Jiannao granules can improve the learning and memory abilities and promote the recovery of cerebral blood flow in the rat model of VCI induced by 2-VO by regulating the expression of proteins involved in the PI3K/Akt signaling pathway, inhibiting inflammation, and reducing hippocampal neuron injury.
3.Kaixin Jieyu Pills Up-Regulated PPARα/FGF21/FGFR1 Pathway to Improve Depression-Like Behavior in Chronic Unpredictable Mild Stress Model Rats
Wujianwen ZHAI ; Fengmei YANG ; Yujing JIN ; Rui YANG ; Shijing HUANG ; Juhua PAN ; Yuxia CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(12):3829-3836
Objective To explore the effects of Kaixin Jieyu Pill(KJP)on peroxisome proliferator-activated receptor alpha(PPARα),serum and hippocampal tissue fibroblast growth factor 21(FGF21)and hippocampal tissue fibroblast growth factor receptor 1(FGFR1)in rats with depression model.Methods A rat chronic stress model was prepared using the chronic unpredictable mild stimulus method(CUMS)to observe behavioral changes in depression.Hippocampal FGF21,FGFR1 and liver PPARα protein and mRNA expression levels were measured in hippocampal tissues by western blot and real-time fluorescence quantification(RT-qPCT).Results The expression levels of PPARα,FGF21 and FGFR1's protein and their mRNA were dramatically decreased in the CUMS model rats compared with the normal group(P<0.001).KJP could modify the depressive-like behaviors of rats(P<0.001),upregulate the protein expression level of PPARα,FGF21,and FGFR1(P<0.05,P<0.01,P<0.01);markedly modify the mRNA expression levels of PPARα,FGF21,and FGFR1(P<0.001,P<0.01,P<0.01).Conclusion PPARα/FGF21/FGFR1 pathway may participate in the pathogenesis of depression;upregulation of PPARα/FGF21/FGFR1 pathway may be related to the antidepressant mechanism of KJP.
4.A Cohort Study of Invigorating Qi and Dissipating Depression Herbs on Depressive Disorders in the Clinical Efficacy
Zehua ZHAO ; Yusong YANG ; Xile YANG ; Shijing HUANG ; Juhua PAN ; Yuxia CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(12):3843-3850
Objective To evaluate the dynamic efficacy of Invigorating Qi and Dissipating Depression Herbs in the treatment of depressive disorders and its effect on recurrence rate and suicidal thoughts.Methods The prospective cohort study was used to divide the patients into three cohorts according to the different exposure factors(treatment regimens):Western medicine group,integrated traditional Chinese and Western medicine group,and Invigorating Qi and Dissipating Depression Herbs group,and followed up for 2 years,and the treatment effects of each cohort at half a year,one year,one and a half years,and two years were observed respectively,and the evaluation indicators were Montgomery Depression Scale(MADRS),Hamilton Anxiety Scale(HAMA)and Social Deficit Screening Scale(SDSS)reduction rates,and endpoint events(relapse,suicidal ideation)incidence;By comparing the efficacy and endpoint event rate of the three cohorts,the effect of Invigorating Qi and Dissipating Depression Herbs on improving depressive disorders was evaluated.Results A total of 409 patients(67 in the Western medicine group,131 in the integrated traditional Chinese and Western medicine group,and 211 cases in the Invigorating Qi and Dissipating Depression Herbs group)were enrolled,and the rate of score reduction after treatment with MADRS,HAMA and SDSS increased compared with the pre-treatment treatment with the extension of the drug taking time.The difference between the MADRS and SDSS reduction rates in the Invigorating Qi and Dissipating Depression Herbs group on taking the drug for half a year and the SDSS reduction rate on one year of taking the drug were statistically significant compared with the Western medicine group,and there was no significant difference compared with the integrated Chinese and Western medicine group.On one and a half years and two years of medication,the integrated Chinese and Western medicine group had more advantages in MADRS reduction rate than the Invigorating Qi and Dissipating Depression Herbs group.The recurrence rates of depression in the three groups within 2 years were 20.9%,30.5%and 36.0%,and the difference was not statistically significant,and the length of taking medication was used as a protective factor by logistic regression analysis.The incidence rates of suicidal thoughts were 44.8%,19.1%and 17.5%,and the differences were statistically significant,and the exposure factors and income level were the protective factors by logistic regression analysis.Conclusion Invigorating Qi and Dissipating Depression Herbs can significantly improve the depressive state of patients,promote the recovery of social function,reduce the occurrence of long-term suicidal ideation,and appropriately extend the time of taking medicine can increase the long-term efficacy and reduce recurrence.
5.Arsenic Content in Cordyceps: A Review
Nuo CHENG ; Juhua PAN ; Jie WANG ; Jun HU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(20):265-273
Cordyceps, a unique precious Chinese herbal medicine in China, has the effects of tonifying the kidney and lungs, stopping bleeding, and resolving phlegm. It has definite clinical effects on diabetes, respiratory diseases (such as chronic obstructive pulmonary disease), chronic kidney diseases, slow arrhythmia, and hyperlipidemia. We searched China National Knowledge Infrastructure (CNKI), Wanfang Data, VIP, SpringerLink (Biomedical Sciences), Embase, Proquest (Life Sciences), PubMed, Cochrane Library, and SinoMed for the research on the arsenic content in Cordyceps. According to statistical analysis, arsenic in Cordyceps mainly exists in the non-toxic or low-toxic form. The arsenic-containing compounds in Cordyceps may be an arsenic-sugar compound with anticancer effect. The inorganic arsenic content in Cordyceps is lower than the arsenic standard stipulated by the State Food and Drug Administration. It remains uncertain whether different measurement methods can affect the results. The total arsenic content varies in Cordyceps from different producing areas, and the general rule of total arsenic content in Cordyceps produced in different provinces, districts, and counties cannot be obtained from the results of the existing studies. The arsenic content in Cordyceps is mainly concentrated in the insect part, which is significantly different from that in stroma, and no arsenic is detected in the mycelia. The soil arsenic content in the growing areas is generally high, which may be a main reason for the high arsenic content in Cordyceps. As a valuable medicinal material for tonifying and nourishing, Cordyceps still plays an irreplaceable role in disease prevention, health care, chronic disease management, and rehabilitation.
6.Meridian Tropism and Compatibility of Chinese Medicine from Theory of Brain Sweat Pore
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(14):204-214
Since LIU Hejian proposed the concept of sweat pore, the theory of sweat pore has experienced accelerated development. Especially with the advances in modern human anatomy and physiology, the microscopic anatomy of sweat pore begins to focus on the intercellular space, ion channels and other membranous space with channels, pores, doors, etc., which exert the functions of exchanging fluid, information, and energy inside and outside blood vessels and discharging metabolic wastes so as to maintain the normal operation of organs. Therefore, sweat pore is the structural basis for the movement of Qi and the central link of Qi-fluid exchange in the body. The brain, as the house of original spirit, is in charge of the spirit of five Zang-organs. The brain sweat pore is pivotal for the circulation of Qi, blood, and fluid in the brain, and it is the structural basis for the normal physiological functions of the brain. The dysfunction of the brain sweat pore will cause the stagnation of Qi and the abnormal transport of blood and fluid. It will cause the abnormal exchange of Qi, liquid and other material and information, which fail to nourish the original spirt and cause the loss of vital activity, eventually leading to consciousness and emotion disorders. The treatment should focus on opening the brain sweat pore, smoothing the exchange of Qi and fluid inside and outside the pore, and restoring the Qi movement, so as to cure encephalopathy. At present, western medicine treatment of encephalopathy needs to solve the problem of drug efflux from the blood-brain barrier and improve the effective concentration of drugs into the brain. The structure and function of brain sweat pore is similar to those of the blood-brain barrier. The aromatic resuscitative medicines and wind-extinguishing medicines can open the brain sweat pore. When being combined with other medicines, they can lead the medicine to enter the brain to restore the Qi movement of the brain sweat pore and enhance the therapeutic effect. Liver-pacifying wind-extinguishing medicines, insect medicines, tonifying medicines, heat-clearing toxin-removing medicines, and damp-draining medicines can treat pathological factors such as wind, phlegm, stasis, deficiency, toxin, and dampness, respectively. These medicines, combined with the medicines with the tropism to brain meridians, can open the brain sweat pore and guide the medicine into the brain to enhance the effective concentration of the medicine, thereby enhancing the efficacy against encephalopathy.
7.Preliminary Conception of Theory of Triple Energizer-nutrient-defense Loop
Yujing JIN ; Jianwen ZHAIWU ; Rui YANG ; Tong ZHANG ; Juhua PAN ; Shijing HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(18):213-224
The research on the essence of triple energizer has not reached a consensus. The correspondence between the existing understanding and the classical theory of triple energizer is still limited in terms of structure and function. According to the traditional theory, nutrient-defense takes channels as the main circulatory system, while the operation of nutrient-defense in the triple energizer remains unclear. Since little is known about the physical structure of the triple energizer, the role of triple energizer as a collection of other Zang-fu organs has been ignored in most cases. The new progress in anatomy paves the way for the research on the essence of triple energizer. The function and structure of triple energizer are similar to those of interstitium and interfacial fluid flow, which enriches our understanding of the macro and micro structures of triple energizer. The triple energizer is distributed throughout the body and composed of membrane and interstitial space. The material structure of triple energizer includes fiber scaffold, collagen fiber, mesenchymal stem cells, histiocytes, pericytes, and interstitial fluid. The functions of triple energizer include passing body fluids, operating nutrient-defense, distributing original Qi, and transmitting and changing pathogenic Qi. According to the available theories and research achievements, we put forward the concept of vertical and horizontal triple energizer, pointed out that triple energizer had independent structure and the features of Zang-fu organs, and preliminarily defined the spatial distribution of triple energizer. The relationship between channels and triple energizer is essential for discussing the operation of nutrient-defense. Telocyte (Tc) and telopod (Tp) has the characteristics of channels in function and structure. The connective tissue with the distribution of Tc and Tp belongs to the same material as the basic structure of interstitial/interfacial fluid flow system and the fibrous skeleton of interstitium. It is clear that channels and triple energizer have material commonality. From the operation paths of nutrient-defense, we proposed that channels may be soaked and attached in triple energizer and put forward the model of channels soaked and attached in triple energizer. By combining the circulation of nutrient-defense with the vertical and horizontal triple energize, we developed the theory of triple energizer-nutrient-defense loop to comprehensively describe the generation, transport, and metabolism of nutrient-defense in channels and triple energizer, aiming to provide a theoretical model for future studies of disease transmission and change from exterior to interior.
8.Preliminary study of the antibody level in confirmed patients with COVID-19 after discharge
Ge SHEN ; Gang YANG ; Ziyan ZENG ; Yan HU ; Qiong LI ; Zugui LIU ; Huiyuan FU ; Junyu HU ; Pan ZHU ; Juhua HUANG ; Qingqing LU ; Shengjie SHI ; Ying HE ; Xiaobing XIE
Chinese Journal of Preventive Medicine 2020;54(12):1448-1452
Objective:To analyze the antibody levels and dynamic changes in patients infected with 2019-novel coronavirus(2019-nCoV).Methods:The average age of 72 corona virus disease 2019 (COVID-19) patients was (45.53±16.74)years(median age:47 year), including (44.88±17.09) years(median age:46 year) for 38 males and (46.32±16.52)years (median age:46 year) for 34 females in Loudi City, Hunan Province. There is no significant difference in genders between the severe and mild groups (χ2=0.916, P>0.05). There is a significant difference in the age between the severe and mild groups ( F=3.315, P<0.05). The blood samples of 72 discharged patients were collected and the consistence of IgM and IgG antibodies were detected by chemiluminescence method. SPSS25.0 was used for gender, age, case type and antibody analysis of variance, χ 2 test and other analysis. Results:The average time of the serum samples collection of 72 patients was (34.89±9.02)days (median time: 34 days) from onset of COVID-19, and (14.53±8.35) days (median time: 14 days) from discharge. The positive rate of IgM or IgG was 97.22% (70/72), and the positive rate of IgM and IgG was 48.61% (35/72) and 97.22% (70/72) respectively. Serum COVID-19 antibodies were detected in 72 patients from 1st to 40th days after discharge. The average concentration of IgM in 1-7 days, 8-14 days, 15-21 days, 22-28 days, above 29 days were 21.91(7.07-52.84)AU/ml, 14.16(6.19-32.88)AU/ml, 11.36(6.65-42.15)AU/ml, 8.15(3.66-30.12)AU/ml, 2.98(0.46-6.37)AU/ml. There was no significant difference in the time of IgM antibody concentration ( H= 8.439, P>0.05). The average concentrations of IgG in 1-7 days, 8-14 days, 15-21 days, 22-28 days, 29 days and above were 169.90 (92.06-190.91) AU/ml, 163.89 (91.19-208.02) AU/ml, 173.31 (95.06-191.28) AU/ml, 122.84 (103.19-188.34) AU/ml, 101.98 (43.75-175.30) AU/ml, respectively, ( H=2.232, P>0.05). The IgM becomes negative after the 3rd week of discharge and decreases rapidly with time. The IgG concentration higher than IgM during the same period, and keep at high level without any change, and decrease in the fourth week. Among them, 5 cases developed "re-infection" within 1-3 weeks after discharge, and the rate of "re-infection" was 6.94% (5/72 cases). Conclusions:After the COVID-19 patients are discharged from the hospital, the level of antibodies produced varies greatly among individuals, but the overall changes in antibodies have a certain pattern. It is recommended to strengthen the antibody monitoring during hospitalization and after discharge from the hospital to reduce the "re-infection" rate and potential risk of infection.
9.Preliminary study of the antibody level in confirmed patients with COVID-19 after discharge
Ge SHEN ; Gang YANG ; Ziyan ZENG ; Yan HU ; Qiong LI ; Zugui LIU ; Huiyuan FU ; Junyu HU ; Pan ZHU ; Juhua HUANG ; Qingqing LU ; Shengjie SHI ; Ying HE ; Xiaobing XIE
Chinese Journal of Preventive Medicine 2020;54(12):1448-1452
Objective:To analyze the antibody levels and dynamic changes in patients infected with 2019-novel coronavirus(2019-nCoV).Methods:The average age of 72 corona virus disease 2019 (COVID-19) patients was (45.53±16.74)years(median age:47 year), including (44.88±17.09) years(median age:46 year) for 38 males and (46.32±16.52)years (median age:46 year) for 34 females in Loudi City, Hunan Province. There is no significant difference in genders between the severe and mild groups (χ2=0.916, P>0.05). There is a significant difference in the age between the severe and mild groups ( F=3.315, P<0.05). The blood samples of 72 discharged patients were collected and the consistence of IgM and IgG antibodies were detected by chemiluminescence method. SPSS25.0 was used for gender, age, case type and antibody analysis of variance, χ 2 test and other analysis. Results:The average time of the serum samples collection of 72 patients was (34.89±9.02)days (median time: 34 days) from onset of COVID-19, and (14.53±8.35) days (median time: 14 days) from discharge. The positive rate of IgM or IgG was 97.22% (70/72), and the positive rate of IgM and IgG was 48.61% (35/72) and 97.22% (70/72) respectively. Serum COVID-19 antibodies were detected in 72 patients from 1st to 40th days after discharge. The average concentration of IgM in 1-7 days, 8-14 days, 15-21 days, 22-28 days, above 29 days were 21.91(7.07-52.84)AU/ml, 14.16(6.19-32.88)AU/ml, 11.36(6.65-42.15)AU/ml, 8.15(3.66-30.12)AU/ml, 2.98(0.46-6.37)AU/ml. There was no significant difference in the time of IgM antibody concentration ( H= 8.439, P>0.05). The average concentrations of IgG in 1-7 days, 8-14 days, 15-21 days, 22-28 days, 29 days and above were 169.90 (92.06-190.91) AU/ml, 163.89 (91.19-208.02) AU/ml, 173.31 (95.06-191.28) AU/ml, 122.84 (103.19-188.34) AU/ml, 101.98 (43.75-175.30) AU/ml, respectively, ( H=2.232, P>0.05). The IgM becomes negative after the 3rd week of discharge and decreases rapidly with time. The IgG concentration higher than IgM during the same period, and keep at high level without any change, and decrease in the fourth week. Among them, 5 cases developed "re-infection" within 1-3 weeks after discharge, and the rate of "re-infection" was 6.94% (5/72 cases). Conclusions:After the COVID-19 patients are discharged from the hospital, the level of antibodies produced varies greatly among individuals, but the overall changes in antibodies have a certain pattern. It is recommended to strengthen the antibody monitoring during hospitalization and after discharge from the hospital to reduce the "re-infection" rate and potential risk of infection.
10.Non-Hodgkin lymphoma with hemophagocytic syndrome as the first manifestation:report of two cases and review of literature
Bin ZHU ; Shaoying PAN ; Juhua ZHU ; Zhiyong DING ; Wenli ZHAO
Journal of Leukemia & Lymphoma 2018;27(2):113-116
Objective To explore the features of non-Hodgkin lymphoma (NHL) with hemophagocytic syndrome (HPS) as the initial manifestation at diagnosis. Methods The morbidity and treatment of two B-cell NHL patients with HPS as the initial manifestation were retrospectively analyzed, and the literature were reviewed at home and abroad. Results Two B-cell NHL patients with HPS were treated with the HLH-94 protocol to control febrile condition. After then, R-CHOP protocol (dexamethasone substituted for prednisone) was used and their results were good. In reviewing the literature, it was felt that the main sub-type of B-cell lymphoma-associated HPS (B-LAHPS) was diffuse large B-cell lymphoma. However, sub-types of T-cell lymphoma-associated HPS (T-LAHPS) had no significant difference. The incidence of T-LAHPS was significantly higher than that of B-LAHPS in China and South Korea, but there was no significant difference in the Western countries. T-cell lymphomas were more vulnerable to HPS. The median age of B-LAHPS patients at diagnosis was significantly higher than that of T-LAHPS patients. The median survival and overall survival time of B-LAHPS patients were longer than those of T-LAHPS patients by virtue of the application of rituximab. Conclusion B-cell NHL with HPS as the initial manifestation treated with the 2004-HLH plus R-CHOP protocol could achieve good outcomes.


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