2020 Vol. 36, No. 3

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Clinical Observation of the Rehabilitation Formula for Banking up Earth to Generate Metal in Treating COVID-19 Patients with Deficiency of Lung and Spleen Syndrome in the Recovery Stage
SHISuo-fang, FANGZhu-yuan, XIONGKan, YEDe-liang, WANGWei-min, CHENYong-chang, WUDi, LIYun-ze, ZHANGYue-qing, CUILei, CHENMing-qi, WANGBo-han
2020, 36(3): 281-285.
[Abstract](707) [PDF 437KB](501)
Abstract:
OBJECTIVE To discuss the influence of the rehabilitation formula for banking up earth to generate metal on COVID-19 patients with deficiency of lung and spleen syndrome in the recovery stage in terms of clinical efficacy and immunology-related indexes. METHODS 120 COVID-19 patients with deficiency of lung and spleen syndrome in the recovery stage were divided into the treatment group (89 cases) and the control group (31 cases). The rehabilitation formula for banking up earth to generate metal was given to the treatment group while the control group was not given any interventions. The course was 14 days. The symptom scores were observed before and after treatment in the two groups and the effective rates in the two groups were compared. RESULTS The effective rate of the treatment group was 94.38%， higher than 54.84% in the control group (P<0.01). The symptom scores before and after treatment in group A and B had no significant differences (P>0.05)， and so did the effective rate (P>0.05). The levels of IgA， Complement C3 after the treatment group were lower than those before treatment (P<0.01). The percentages of CD3， CD4 and CD8 were higher after the treatment group than those before (P<0.01). Compared with those in the control group， the percentages of CD3 and CD4 in the treatment group had significant difference(P<0.05). CONCLUSION The rehabilitation formula for banking up earth to generate metal can effectively improve the clinical symptoms caused by deficiency of lung and spleen for COVID-19 patients in the recovery stage as well as their immunologic function.
Protection Strategies for COVID-19 Based on the Theory of Preventative Treatment
ZHANGHao-wen, HEFei-yu, CAIYun, TUYue, HUANGYing, LIUHua-dong, GUYi-huang, CHENDi-ping
2020, 36(3): 286-289.
[Abstract](551) [PDF 442KB](395)
Abstract:
The theory of preventative treatment， the core ideology in traditional Chinese medicine， can be applied into every prevention stage of COVID-19. In terms of three prevention stages of COVID-19， the study found that aromatic medicines can be prescribed to dispel filth before its onset， resulting in protecting the lung and kidney as well as nourishing healthy qi. As for the stage of controlling the development of disease， we should pay much attention to the characteristics of dampness toxin and avoid transforming into heat or stasis， or even worse into block or collapse pattern. When it comes to the stage of preventing recurrence， the principle of strengthening earth to generate metal and recovering both the body and mind should be followed. In addition， balanced diet， life-cultivation qi gong， music， and massage on acupoints can be applied to truncate the risk of recurrence and promote the rehabilitation， with a view to providing thoughts for the prevention of COVID-19.
The Academic Insight of SARS-CoV-2 in Traditional Chinese Medicine from Treatise on Warm Pestilence<\i>
ZHANGJian-bin, SHENJia, XIONGJia-wei, CHENMei-yun
2020, 36(3): 290-294.
[Abstract](657) [PDF 448KB](367)
Abstract:
The paper discussed the naming， characteristic， focus， transmission， diagnosis and treatment of COVID-19 based on Treatise on Warm Pestilence. It was suggested that the data related to COVID-19 could be collected to disclose its transmission process in terms of pathogenic characteristic， further explore its diagnosis and treatment modes with regard to the interaction of evil qi and healthy qi， and elaborate the modern significance of latent pathogen and pleurodiaphragmatic interspace (membrane-source) based on the research results in modern medicine and the theory of latent pathogen in the pleurodiaphragmatic interspace. The academic insight of doctrine of epidemic febrile disease in traditional Chinese medicine (TCM) has been further developed and improved through the study on Treatise on Warm Pestilence， epidemiologic analysis， clinical manifestation， laboratory inspection， along with TCM diagnosis and treatment.
Network Pharmacology of Jinxin Oral Liquid in the Treatment of COVID-19
HEYu, XUHai-yan, SHANJin-jun, ZHAOXia, SHENCun-si, JIJian-jian, LINLi-li, YANGRui, XIETong, WANGShou-chuan
2020, 36(3): 295-299.
[Abstract](536) [PDF 3457KB](391)
Abstract:
OBJECTIVE The network pharmacology was used to explore the targets of Jinxin oral Liquid in the treatment of COVID-19. METHODS TCMSP database was used to mining the drug-like components in Jinxin oral Liquid and corresponding targets. The targets of pneumonia-related diseases in DisGeNET library was pointed and matched to the components' targets. STRING database was used for protein interaction network diagram. The DAVID website was used for further gene ontology analysis. RESULTS A total of 129 drug like compounds were obtained. Totally， 75 gene targets were matched to drug like components and pneumonia-related diseases. Protein interaction and gene ontology analysis suggested that the antiviral immune response and anti-inflammatory responses are most enriched. CONCLUSION The components in Jinxin oral Liquid were the possible candidates for treatment of COVID-19.
Discussion on Preventive and Therapeutic Principle of Radiation Injury Based on the Theory of Stasis and Heat
WUMian-hua, WUYan, LIWen-ting
2020, 36(3): 300-302.
[Abstract](592) [PDF 366KB](405)
Abstract:
Along with the development of nuclear medicine， radiation therapy is very important in the comprehensive treatment of tumor， but radiation damage is the main bottleneck restricting its development. National traditional Chinese medicine (TCM) master Professor Zhou Zhongying held that the etiology and pathogenesis of radiation injury was fighting of static blood and extreme heat， as well as deficiency of qi and yin under the guidance of the theory of stasis and heat. Radiation injury was characterized by burning pain， delayed traumatic， damaged collateral， impaired yin and qi. In TCM， the focus of it is usually classified by its location in head and neck， chest， epigastrium and hypogastrium. When it came to different foci， the clinical manifestation would be different. Hence， Professor Zhou regarded cooling blood and dispersing stasis along with nourishing yin and invigorating qi as its preventive and therapeutic principle.
Discussion on Traditional Chinese Medicine in Tumor Prevention and Treatment by Master Zhou Zhongying
LI Liu, CHENG Hai-bo, YE Fang, WU Mian-hua, ZHOU Zhong-ying
2020, 36(3): 303-306.
[Abstract](570) [PDF 435KB](417)
Abstract:
Based on the current situation of traditional Chinese medicine in the prevention and treatment of tumor， Professor Zhou Zhongying puts forward eight important arguments，which include careful using of attack medicine to prevent pathogenic decline and positive consumption，using detoxification and poison attack medicine with different primary and secondary， paying attention to individuals with precise treatment， reducing toxicity and increasing efficiency with multiple methods of integration，reversing pathological changes from evil to good，using follow-up treatment to prevent relapse，increasing confidence by psychosomatic care，giving diet suggestion varies from person to person. The purpose is to further provide guidance for the clinical diagnosis and treatment of tumor in traditional Chinese Medicine.
Quantitative Assessment of CMR T1 Mapping on the Interventional Effects of Wenlü Pingji Granule on Myocardial Fibrosis and Atrial Fibrillation
WANGXin-dong, SUNXue-mei, FANGZhu-yuan
2020, 36(3): 307-312.
[Abstract](445) [PDF 615KB](346)
Abstract:
OBJECTIVE To observe the quantitative assessment of cardiac magnetic resonance longitudinal relaxation time (CMR T1 mapping) on the interventional effect of Wenlü Pingji Granule on patients with atrial fibrillation (deficiency of both qi and yin) in terms of left ventricular myocardial fibrosis. METHODS 43 patients with atrial fibrillation with deficiency of qi and yin syndrome who met the inclusion criteria from March 2017 to November 2019， including 17 cases of persistent atrial fibrillation and 26 cases of paroxysmal atrial fibrillation. They were randomly divided into western medicine control group (20 cases) and Chinese medicine group (23 cases) applied with Wenlü Pingji Granule. The treatment and observation period was 24 weeks. The left ventricular myocardial fibrosis after treatment was quantitatively assessed by CMR T1 mapping imaging. ELISA was applied to detect serum interleukin 18 (IL-18)， tumor necrosis factor-α (TNF-α)， superoxide dismutase (SOD)， malondialdehyde (MDA)， liver kinase B1 (LKB1) levels and fibrosis indicator matrix metalloproteinase-2 (MMP-2)， pre-collagen amino terminal peptide Ⅲ (PⅢ NP) so as to assess the changes of upstream oxidative inflammation-related factors in atrial fibrillation fibrosis. RESULTS The number of recurrence frequency and rate of atrial fibrillation in patients with paroxysmal atrial fibrillation in the Chinese medicine group were lower than those in the control group (P<0.05). The T1 value and ECV were significantly decreased (P<0.01)， serum MMP-2， PⅢ NP， IL-18， TNF-α and MDA levels were significantly decreased (P<0.01) and SOD and LKB1 levels were significantly increased (P<0.01) after treatment in the Chinese medicine group， which were better than those in the western medicine control group (P<0.01). CONCLUSION Wenlü Pingji Granule can keep Sinus rhythm maintenance and reduce recurrence in patients with paroxysmal atrial fibrillation， as well as inhibit myocardial fibrosis in atrial fibrillation patients with deficiency of qi and yin syndrome. The mechanism may be related to the inhibition of the upstream oxidative inflammatory cascade.
Study on the Correlation Between Single-Center Adult Urinary Calculi and Traditional Chinese Medicine Constitution
YU Qiu, LU Zi-jie, CHEN Xue-hua, MA Long, ZHOU Yue, XU Yan, GU Xiao-jian, ZHANG Li
2020, 36(3): 313-317.
[Abstract](428) [PDF 424KB](354)
Abstract:
OBJECTIVE To explore the correlation between adult urinary calculi and traditional Chinese medicine (TCM) constitution， and to provide new directions and strategies for the prevention and treatment of urolithiasis. METHODS A total of 998 adult patients with urinary calculus in Jiangsu Province Hospital of Chinese Medicine from June 2018 to December 2019 were recruited. The basic information such as sex， age， height， body weight， location of disease， recurrence， analyzed the results of calculus composition and identified the TCM constitution types were collected. The statistical methods were applied to analyze the correlation between adult urinary calculus and TCM constitution types. RESULTS Adult urinary calculus patients' TCM constitution types were significantly correlated with their BMI， gender， age and location (P<0.01). The most common types of TCM constitutions in adult patients with urinary calculus were phlegm-dampness， dampness-heat， yang deficiency and yin deficiency， among which male patients were mainly dampness-heat and phlegm-dampness while female patients were mainly yang deficiency and phlegm-dampness. A total of 998 adult patients with urinary calculus were found to be composed of 23 kinds of calculus composition， including 334 of single ones， 462 of two compound ones， 194 of three composition compound ones， 8 of four composition compound ones. The mixed form was more common in clinics. The affected patients were mainly middle-aged people， and their constitutions were mostly phlegm-dampness and yang deficiency. The young patients with urinary calculus were mostly dampness-heat and the elderly patients yin deficiency. The main focus located in the upper urinary tract. Calcium oxalate monohydrate， calcium oxalate dihydrate， and carbonate apatite were the most common calculus components. Qi deficiency was more likely to cause infectious calculi in adult patients than the type of dampness-heat， phlegm-dampness and yin deficiency (P<0.003). Patients with dampness-heat had a higher recurrence rate of calculus than patients with other constitutions. Mild constitution patients were not so likely to relapse (P<0.003). CONCLUSION There is a clear correlation between adult urinary calculus and TCM constitution. From the perspective of Chinese medicine constitution， it provides new methods for clinical prevention and treatment of adult urinary calculus.
Clinical Research of Curcumin Microemulsion Gel on Skin Fibrosis after Radiotherapy
HANGui-xiang, LIUJia, MAChao-qun, ZHUXuan-xuan, ZHANGDi
2020, 36(3): 318-321.
[Abstract](553) [PDF 530KB](366)
Abstract:
OBJECTIVE To explore the curative effect of curcumin microemulsion gel on skin fibrosis after radiotherapy and the effect for fibrosis index factors. METHODS 80 patients with skin fibrosis diagnosed after tumor radiotherapy from March 2017 to December 2018 in the Department of Oncology of Affiliated Hospital of Nanjing university of Chinese medicine were selected. Curcumin microemulsion gel was set as the treatment group and asiaticoside cream was set as the control group， with 2 times daily skin administration for 2 consecutive months. The total effective rate， western medicine score， traditional Chinese medicine (TCM) syndrome score and expression of fibrosis factor in the two groups were compared. RESULTS After treatment， the total effective rate of the treatment group was 92.5%， better than 77.5% in the control group(P<0.05). Two groups of western medicine treatment efficacy scores were significantly improved (P<0.05). The treatment group was significantly better than the control group in improving the arthralgia (P<0.05). 2 groups of TCM syndrome evaluations were improved (P<0.05). The skin of the treatment group was hard and the symptoms of limb numbness were better than those of the control group (P<0.05). The serum fibrinization factor hydroxyproline (HYP)， transforming growth factor-β (TGF-β) and connective tissue growth factor (CTGF) of the two groups all decreased significantly (P<0.01). The contents of TGF-β and CTGF in blood serum of treatment group were significantly lower than those of control group (P<0.05). CONCLUSION Curcumin microemulsion gel treatment has significant effects on skin fibrosis after radiotherapy， and reduces the levels of fibrosis factor in patients' blood significantly.
Observation on the Efficacy of Qingli Xiezhuo Qutong Decoction Combined with Western Medicine for the Treatment of Acute Gouty Arthritis with Damp-Heat Accumulation Syndrome
NIU Chao-yang, LI Peng-chao, MENG Qing-liang
2020, 36(3): 322-325.
[Abstract](488) [PDF 403KB](396)
Abstract:
OBJECTIVE To observe the clinical efficacy of Qingli Xiezhuo Qutong Decoction in combination with Western medicine for the treatment of acute gouty arthritis with damp-heat accumulation syndrome. METHODS 70 patients were randomly divided into the control group and the treatment groups with 35 cases each. The control group was given colchicine tablets orally at an initial dose of 1 mg， followed by 0.5 mg per hour and 0.5 mg at a time， for 3 times per day when symptom was relieved， and the treatment group was given Qingli Xiezhuo Qutong Decoction one dose a day. Both groups were treated for 7 days. The joint pain score， joint swelling score， and traditional Chinese medicine (TCM) syndrome score were recorded before and after treatment， and blood leukocyte (WBC)， erythrocyte sedimentation rate (ESR)， C-reactive protein (CRP)， blood uric acid (BUA)， tumor necrosis factor-α (TNF-α)， interleukin-1 (IL-1) levels were tested before and after treatment， and safety evaluation was conducted. RESULTS The clinical efficacy of the treatment group was better than that of the control group after treatment (P<0.05). Joint pain score， joint swelling score， TCM syndrome score and CRP， ESA， TNF-α， IL-1， BUA levels were significantly reduced in both groups (P<0.01) and the treatment group was better than the control group (P<0.05， P<0.01). CONCLUSION On the basis of colchicine， Qingli Xiezhuo Qutong Decoction for acute gouty arthritis (damp-heat accumulation syndrome) can effectively relieve the main clinical symptoms reducing the inflammatory response， with better safety and clinical efficacy than that of pure western medicine.
Effects of Huazhuo Jiedu Formula on Chronic Atrophic Gastritis Patients with Helicobacter Pylori<\i> Infection
BAIHai-yan, HAOXu-rui, LINa, LIUKai-juan, LIWei-kang, YANGQian
2020, 36(3): 326-330.
[Abstract](520) [PDF 448KB](366)
Abstract:
OBJECTIVE To observe the Helicobacter pylori(Hp) eradication rate and clinical effect of Huazhuo Jiedu Formula in the treatment of chronic atrophic gastritis (CAG) with Hp infection， and to explore its possible mechanism. METHODS 120 patients of CAG with Hp infection were randomly divided into two groups: the treatment group and the control group. The treatment group was given one dose of Huazhuo Jiedu formula per day， while the control group was given standard quadruple bactericidal therapy for 2 weeks， followed by placebo， one dose per day. After 3 months' treatment， TCM syndrome score， gastroscope mucosal symptom score， pathological tissue change score， gastrin-17 (G-17)， serum pepsinogen Ⅰ， Ⅱ (PGⅠ， PGⅡ)， PGⅠ/PG Ⅱ ratio (PGR)， cytotoxin associated protein (CagA)， vacuole cytotoxin (VacA) and urease B (UreB) of the two groups were compared before and after treatment. The eradication rate of HP was recorded before treatment， 6 weeks and 3 months after treatment， and after the follow-up for 6 months to observe the recurrence of Hp. RESULTS After three months' treatment， TCM syndrome scores in the two groups were significantly lower than those before treatment with significant difference (P<0.05， P<0.01)， compared with the control group (P<0.05， P<0.01). Compared with the control group， the gastroscope mucosal symptom score and pathological tissue change score in the treatment group were lower than those before treatment (P<0.05， P<0.01)， compared with the control group (P<0.05， P<0.01). The levels of PGⅠ and PGR in the treatment group were higher than those before treatment， and the levels of PG Ⅱ and G-17 were lower than those before treatment (P<0.05， P<0.01)， compared with the control group (P<0.05， P<0.01). The positive rates of CagA， VacA and UreB in the two groups were significantly lower than those before treatment (P<0.01)， compared with the control group (P<0.05， P<0.01). The eradication rate of Hp of patients in the treatment group was better than that in the control group (P<0.05). The 6 months follow-up recurrence rate of the treatment group was lower than that of the control group (P<0.01). CONCLUSION Huazhuo Jiedu formula， having good effect on Hp inhibition and low recurrence rate， is effective in the treatment of CAG with Hp infection. It can effectively alleviate clinical symptoms， improve pathological changes and promote mucosal repair. Its mechanism may be related to the levels of CagA， VacA， UreB， PGⅠ， PGⅡ， PGR and G-17.
Bacteriostasis of Prenylated Flavonoids from Sophora Flavescens<\i> and Flavonoids from Glycyrrhiza Uralensis<\i> Alone and Their Combination on Staphylococcus Aureus<\i> and Experimental Mastitis
CAOYa-qi, WEIDan-dan, ZHANGSen, ZENGFei, GUOSheng, GUOJian-ming, DUANJin-ao
2020, 36(3): 331-338.
[Abstract](593) [PDF 745KB](386)
Abstract:
OBJECTIVE To explore the antibacterial activity of prenylated flavonoids from Sophora flavecens and licorice flavonoids and their combination in vitro and in vivo， so as to provide scientific basis for the resource value discovery and comprehensive development of Sophora flavescens Ait. and Glycyrrhiza uralensis Fisch. METHODS The UPLC-Q-TOF-MS technology was applied to qualitatively analyze the prepared prenylated flavonoids from Sophora flavescens Ait. (KSHT) and licorice flavonoids (GCHT). The in vitro antibacterial activity was evaluated by investigating the effect of a single use and combined use of KSHT and GCHT on inhibiting the growth of Staphylococcus aureus and the generation of biofilm and flavin. The in vivo antibacterial activity was evaluated by building a mouse mastitis model induced by Staphylococcus aureus. RESULTS A total of 20 and 22 flavonoids were identified from KSHT and GCHT， respectively. KSHT outperformed GCHT in inhibiting the growth of Staphylococcus aureus and the generation of biofilm and flavin. Moreover， the combined use of KSHT and GCHT had better inhibitory effect than their independent use， suggesting that they have synergistic antibacterial action. Whether they were independently or jointly used in high or low doses， all the indexes of mastitis in mice were improved. Compared with the independent use of KSHT and GCHT， the combined use can better improve the viscera index and fungal burden of tissue in mice with mastitis. Regarding the improvement of indexes， KSHT was better than GCHT. In terms of the effect on improving the histopathology of mammary gland and reducing IL-1β， IL-2 and TNF-α in model group， GCHT was better than KSHT， but equivalent to the combination group. CONCLUSION The results of in vitro and in vivo tests indicated that the combined use of KSHT and GCHT has the best effect in resisting Staphylococcus aureus. It is expected that they can be jointly used in the prevention and treatment of mastitis in animal husbandry.
Investigation on Protective Effects of Medicated Sera of Sinisan Containing Bupleuri Radix and Paeoniae Radix Alba before and after Vinegar-Processing on CORT-Induced PC12 Cell Injury
LIUJing, CAIHao, DUANYu, PEIKe, ZHOUJia, ZHANGYa-ting, MOZi-qing, NIUMin-jie
2020, 36(3): 339-345.
[Abstract](498) [PDF 1235KB](340)
Abstract:
OBJECTIVE To investigate the molecular mechanism of anti-depression and protective effects of medicated sera of Sinisan containing Bupleuri Radix and Paeoniae Radix Alba before and after vinegar-processing on CORT-induced PC12 cell injury. METHEODS Firstly， the MTT and morphological observation methods were used to evaluate whether the model was established successfully. Secondly， the cultured cells were divided into four groups (blank control group， CORT group， SNS serum group， VPSNS serum group). ELISA method was used to determine the contents of the monoamine neurotransmitters (DA and 5-HT) and the inflammatory cytokines (TNF-α and IL-1β) in cell supernatant. Then， the content of the injury index (LDH) and the activity of the protective index (SOD) in cell supernatant were measured using the microplate method， and the cell apoptosis rates were measured by flow cytometric analysis. Finally， the protein and gene expression of BDNF， ERK， and CREB were detected by Western blot and qPCR. RESULTS Compared to the model group， both 20%SNS serum group and 20%VPSNS serum group increased the contents of DA， 5-HT， the activity of SOD， and the protein and gene expression of BDNF， ERK， and CREB (P<0.05). And both of them reduced the contents of TNF-α， IL-1β， LDH， and the cell apoptosis rates (P<0.05). The protective effect of 20%VPSNS serum group on CORT-induced PC12 cell injury was better than that of 20%SNS serum group (P<0.05). CONCLUSION The BDNF-ERK-CREB pathway is probably involved in the protective effects of SNS and VPSNS on CORT-induced PC12 cell injury， and the protective effect of VPSNS is better than that of SNS.
Study on the Anti-Apoptosis of Ferulic Acid on Osteoblast and Its Correlation with GPR30
HUANGTian-yi, CHENTing-ting, CUIJie, LIMeng-yu, HUAYong-qing
2020, 36(3): 346-351.
[Abstract](514) [PDF 1581KB](342)
Abstract:
OBJECTIVE To investigate the effects of ferulic acid (FA) on osteoblast apoptosis and its mechanism， and explore the potential therapeutic effects of FA on osteoporosis. METHEODS The pre-osteoblast cell line MC3T3-E1 was selected. The effect of FA on cell viability was observed by MTT assay. Apoptosis was detected by flow cytometry. The expression of apoptosis-related protein Bcl-2 was detected by Western blot. Flow cytometry and DCFH-DA fluorescence staining were performed to observe the reactive oxygen species (ROS) level. The nuclear localization of FoxO3a， one member in the forkhead box (Fox) family， was observed by immunofluorescence. The G protein coupled receptor 30 (GPR30) specific antagonist G15 was used to observe FA effect. RESULTS FA showed no significant effect on normal cells in the dose range of 25~400 μmol/L， but promoted the cell viability after hydrogen peroxide injury. Flow results showed that FA could reduce cell apoptosis rate and ROS level after peroxidation injury. Immunofluorescence results indicated that FA could inhibit the apoptosis and the nuclear content of oxidative stress-related protein FoxO3a under peroxidative conditions. After treatment with G15， the effects of FA on decreasing apoptosis， lowering ROS levels， and reducing the content of FoxO3a in the nucleus were significantly blocked. CONCLUSION FA shows the ability to resist osteoblast apoptosis， reduce intracellular ROS levels， and prevent FoxO3a nuclear translocation under conditions of peroxidative damage. The mechanism may be related to the membrane estrogen receptor GPR30.
Observation on the Effect of Erzhiwan on Zebrafish Osteoporosis Model and Study on Autophagy Mechanism of Osteoclast
TIANTao, FUQiang, ZHUJian-kui, YINHong
2020, 36(3): 352-357.
[Abstract](616) [PDF 3170KB](354)
Abstract:
OBJECTIVE To observe the treatment efficacy of Erzhiwan on prednisolone-induced osteoporosis of zebrafish model and to clarify its regulation of autophagy mechanism on osteoclast differentiation. METHODS The young zebrafishes of good energy after 3 days， normal development were selected. The zebrafish osteoporosis model was established with 25 μmol/L prednisolone. The zebrafishes were divided into control group， model group， etidronate disodium group (300 mg/L)， Mohanlian group (50 μg/mL)， Nvzhenzi group (50 μg/mL) and Erzhiwan group (100 μg/mL). After 4 days， the calcein staining was adopted to measure the vertebral bone fluorescence area of zebrafish; qPCR was adopted to detect the osteogenic marker genes ALP， BMP-2b， Runx2 and osteoclast marker genes CTSK， TRAP， NFATC-1 expression. RAW264.7 cells were divided into control， LPS induction (10 μg/L)， EZW(100 μg /mL)， MHL(50 μg/mL)， and NZZ group (50 μg/mL) after they were cultured to 80%~90% density. The mRNA expression of TRAP， CTSK and autophagy-related genes 5 (ATG5)， ubiquitin-binding protein 62 (p62)， mammalian target of rapamycin(mTOR)，and Beclin were detected by qPCR. RESULTS Compared with the control group， the model group had significant declines in both vertebral bone fluorescence area and osteogenic marker gene expression and had significant rise in osteoclast marker genes（P<0.05，P<0.01）. Compared with the model group， the ED group， the Nvzhenzi group and the Erzhiwan group all remarkably improved zebrafish osteoporosis and acted better than both Mohanlian group and Nvzhenzi group（P<0.01）. qPCR showed that compared with the control group， the model group's osteogenic marker gene expression significantly declined and osteoclast marker genes rose conspicuously（P<0.05，P<0.01）. Compared with model group， the ED group， Mohanlian group and the Nvzhenzi group as well as Erzhiwan group had osteogenic marker gene expression rose significantly（P<0.05，P<0.01）， and Erzhiwan group has osteoclast marker genes reduced significantly（P<0.01）. The in-vitro study showed that the expression of osteoclast marker genes CTSK， TRAP， ATG5 and NFATC-1 induced by LPS as well as autophagy-related genes mTOR， ATG5， Beclin， p62 all rose（P<0.01）. The Erzhiwan group， Mohanlian group and Nvzhenzi group significantly decreased the expressions of osteoclast marker genes and autophagy-related genes（P<0.05， P<0.01）. CONCLUSION Erzhiwan can significantly improve the clinical symptoms of osteoporosis in zebrafish model， its mechanism may be related to the inhibition of the osteoclast-differentiation by reducing autophagy-related gene expressions.
Mechanism of Jinshui Liujun Decoction in the Treatment of Chronic Obstructive Pulmonary Disease Based on Network Pharmacology
YUE Qin-fei, CHEN Ye-qing, FAN Xin-sheng
2020, 36(3): 358-364.
[Abstract](545) [PDF 716KB](359)
Abstract:
OBJECTIVE To explore the mechanism of Jinshui Liujun decoction in the treatment of chronic obstructive pulmonary disease (COPD) by network pharmacology. METHODS The chemical ingredients and corresponding targets of Jinshui Liujun decoction were retrieved from TCMSP platform. The relevant targets of COPD were obtained through the TTD， DisGeNET， GeneCardsand and CTD databases， the targets of drug chemical ingredients were mapped to the targets of COPD. The intersection targets and the efficacy components were obtained. Using STRING database and Cytoscape 3.7.2 software， component-target network model was constructed. The OmicShare open platform was used to perform gene function (GO) and the Kyoto Gene and Genome Encyclopedia (KEGG) enrichment analysis. RESULTS Jinshui Liujun decoction had 96 intersection targets for COPD， 435 efficacy components (The main components are quercetin， stigmasterol， kaempferol， β-sitosterol， naringin， ursolic acid， etc.)， and 20 key targets which degree，betweenness and closeness are greater than average， including MAPK3， TNF， PPAR-γ， SIRTI， HMOX1， MAPK8， CAT， SIRT1， STAT3， etc. Enrichment analysis showed that the formula mainly involved the response to biological and chemical stimuli， various cell biological processes， cell proliferation， immunity， metabolism， etc. 17 signaling pathways including IL-17， Toll-like receptor， TNF， HIF-1， FoxO， TGF-β and ErbB， etc. play a role in the treatment of COPD. CONCLUSION Jinshui Liujun decoction can treat COPD through the synergy of multiple ingredients， multiple targets and multiple pathways， which provides basic data for in-depth exploration of the mechanism of Jinshui Liu jun decoction in the treatment of COPD.
Effect of Hedan Tiaozhi Recipe on Lipid Metabolites in Mice
CAILin-ling, FANGLi-wen, LIUZi-xiu
2020, 36(3): 365-369.
[Abstract](503) [PDF 754KB](342)
Abstract:
OBJECTIVE To study the regulatory effect of Hedan Tiaozhi Recipe on abnormal plasma lipid metabolism in hyperlipidemia mice. METHODS By using UPLC-LTQ/Orbitrap XL， the effect of drug intervention on lipid metabolizer group was investigated， potential biomarkers were searched， and the action way of Hedan Tiaozhi Recipe in regulating lipid metabolism was explored. RESULTS Mass spectrometry of plasma lipids of normal group， model group and Hedan Tiaozhi Recipe group were all significantly separated. 11 biomarkers were identified in the plasma， mainly including phosphatidylcholine， lysophosphatidylcholine and triglycerideincluding phosphatidylcholine. The intervention of Hedan Tiaozhi Recipe significantly improved some abnormal lipid metabolism. CONCLUSION Hedan Tiaozhi Recipe can effectively alleviate the metabolism disorder of phosphatidylcholine， lysophosphatidylcholine and triglyceride induced by high-fat diet， which indicated that the mechanism of Hedan capsule regulating blood lipid may be related to the regulation of these metabolic pathways.
Effect of Zuogui Jiangtang Shuxin Formula Medicated Plasma on Foaming and Apoptosis of Macrophage Induced by ox-LDL Through PERK/ eIF2α/ATF4 Signaling Pathway
YANGJin-wei, ZHAOCan, LIUXiu, WUYong-jun, YURong
2020, 36(3): 370-375.
[Abstract](541) [PDF 2717KB](367)
Abstract:
OBJECTIVE To investigate the effect of Zuogui Jiangtang Shuxin Formula medicated plasma on ox-LDL-induced PERK/eIF2α/ATF4 integrated stress response and activation of apoptosis key signaling molecules CHOP and Caspase-12. METHEOD J774A.1 cells were randomly divided into 5 groups: normal group， model group， TUDCA group， Zuogui Jiangtang Shuxin Formula medicated plasma group and western medicine combination (metformin+atorvastatin calcium) medicated plasma group. All the drug intervention groups were pretreated with ox-LDL to establish an experimental model for macrophage foam cell formation. The cells of drug intervention groups were cultured in 10% different drug containing plasma， respectively. Cells of normal and model groups were cultured in normal culture medium. The growth inhibition of J774A.1 were tested by CCK-8 essay after 24 h culture. Oil Red O Dyeing experiment was used to show the cellular lipid droplets. The expression of p-PERK， p-eIF2α， ATF4， CHOP and Caspase-12 were detected by Western blot. RESULTS 100 mg/L ox-LDL suppressed the growth of J774A.1 cells with an inhibitory rate of (50.06±0.09)%. Oil Red O staining showed that the model group cells had more lipid droplets than normal group (P<0.01). Drug intervention groups effectively prevented the pervasion of ox-LDL to reduce lipid accumulation. Oil red O staining showed that lipid aggregated in Zuogui Jiangtang Shuxin Formula medicated plasma， compared with western medicine combination medicated plasma group (P<0.01)， and the effect was equivalent to TUDCA group (P＞0.05). ox-LDL up-regulated the expression of p-PERK， p-eIF2α， ATF4， CHOP and Caspase-12. Zuogui Jiangtang Shuxin Formula medicated plasma significantly restrained the expression of p-PERK， p-eIF2α and ATF4， and apparently decreased endoplasmic reticulum associated apoptosis protein CHOP and Caspase-12 expression (P<0.01). There was no significant difference in Zuogui Jiangtang Shuxin Formula medicated plasma group and TUDCA group. And Zuogui Jiangtang Shuxin Formula medicated plasma group was superior to the western medicine combination medicated plasma group (P<0.01). CONCLUSION Zuogui Jiangtang Shuxin Formula medicated plasma can suppress foam cell formation and apoptosis in J774A.1 cells， which may be related to the regulation of PERK/eIF2α/ATF4 signaling pathway and the expression of CHOP and Caspase-12.
Effects of Guanxinping-Containing Serum on RAW 264.7 Macrophages Polarization Induced by LPS
ZHOUGuan-jin, SUNYu-ting, ZHOULi-yu, CHENWei-kai, ZHANGLin-hui, HUWen-qi, LIUFu-ming, YUANDong-ping
2020, 36(3): 376-379.
[Abstract](778) [PDF 598KB](388)
Abstract:
OBJECTIVE To investigate the effects of Guanxinping (GXP) on RAW264.7 macrophage polarization induced by LPS. METHODS Macrophages were divided into control group， model group and low-， mid-， high-dose of GXP groups. Cells were starved for 24 hours before intervention with drug-containing serum of rats. The characteristic molecules of macrophage polarization in cell supernatant， including tumor necrosis factor α (TNF-α) and transforming growth factor β(TGF-β) were detected by ELISA; the expression levels of M1 and M2 macrophage surface molecules CD86 and CD206 were detected by flow cytometry; the expression of mRNA of CD86 and CD206 were detected by qPCR. RESULTS Compared to the model group， high-dose of GXP decreased the secretion of TNF-α (P<0.01); low-， mid-， high-dose of GXP decreased the secretion of TGF-β1 (P<0.01) in macrophage supernatant. Low- and high- dose of GXP decreased the expressions of CD86 (P<0.05) and increased the expression of CD206 (P<0.01). There were no significant difference in the mRNA expression of CD86 between the GXP groups and model group， but GXP increased the mRNA expression of CD206. CONCLUSION GXP can inhibit the polarization of macrophages towards M2， which may be one of the mechanisms of GXP in the treatment of atherosclerosis.
Effects of Luteolin and Its Analogues Against Myocardial Ischemia/Reperfusion Injury， Lipid Metabolism and Apoptosis
ZOUXiao-yan, FANXiang-cheng, TIANYou-qing, SHANGJing
2020, 36(3): 380-386.
[Abstract](522) [PDF 1356KB](392)
Abstract:
OBJECTIVE To explore the multi-component and on MIRI model rat multi-target mechanism of Chinese medicine against MIRI， the protective effects of luteolin， kaempferol and luteoloside on rat MIRI， and the inhibition on lipid-metabolization-related enzymes 11β-HSD1， LPAATβ， and apoptosis-related enzymes PTEN， and SHP-2. METHEODS Using a Langendorff apparatus， the MIRI model was established by stopping perfusion and reperfusion with oxygenated Locke-Ringer's solution. The HR， LVEDP， LVDP， (-dp/dt)/(+dp/dt)， CF， LDH and myocardial infarct size were assayed. LPAATβ and 11β-HSD1 were assayed by spectrophotometry. The inhibition of PTEN and SHP-2 activity was measured by spectrophotometry. RESULTS Luteolin， kaempferol and luteoloside could protect HR， reduce LDH release， raise LVDP， LVEDP and (-dp/dt)/ (+dp/dt)， recover CF， and reduce myocardial infarct size， respectively. Moreover， the activities of LPAATβ， 11β-HSD1， PTEN and SHP-2 were repressed by pretreatment with luteolin， kaempferol and luteoloside. CONCLUSION Luteolin， kaempferol and luteoloside show the cardioprotective effects on MIRI of isolated rat heart by against lipid metabolism disorders and apoptosis.
Study on Variation of Non-Volatile Compounds of Raw Ginger after Processing by UHPLC-Orbitrap/HRMS
CHENChen, GENGJian-liang, KUANGHai-xue, WANGQiu-hong
2020, 36(3): 387-395.
[Abstract](539) [PDF 747KB](343)
Abstract:
OBJECTIVE To study the different non-volatile compounds between raw ginger and processed ginger by UHPLC-Orbitrap/HRMS. METHEODS Raw ginger and processed ginger were extracted by 80% methanol， respectively. The characteristic compounds were identified by retention time， accurate molecular weight and MS fragmentation. The different compounds were screened by multivariate analysis and their bioactivities were measured. RESULTS Seventy-two compounds were identified or tentatively characterized. The combination of principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) revealed the different chemical profiles of raw ginger and processed ginger and 11 compounds， including 6-shogaol， 8-shogaol， 10-shogaol， 12-shogaol， 6-gingerol， 10-gingerol， 6-paradol， 6-dehydrogingerdione， 8-dehydrogingerdione， methylacetoxy-6-gingerdiol and 7-(4-hydroxyphenyl)-1-phenylhept-4-en-3-one were found changed after processing. Among those compounds， shogaols showed significant anti-inflammatory activities. CONCLUSION The results provide evidence to explain the efficacy differences between raw ginger and processed ginger as well as the processing mechanism.
Enrichment of Thiol-Containing Peptides in Water Buffalo Horn by Core-Shell Fe3O4@PDA@Au nano-Materials
LIUTing, HANShu-ying, KANGAn, DENGHai-shan, ZHUDong, CHIYu-mei, LIURui
2020, 36(3): 396-400.
[Abstract](476) [PDF 1568KB](342)
Abstract:
OBJECTIVE Enrichment， identification， and characterization of thiol-containing peptides in water buffalo horn by using core-shell Fe3O4@PDA@Au nano-material. METHODS The core-shell Fe3O4@PDA@Au magnetic nano-materials were prepared by seed growth method， and the material was applied to the enrichment of thiol-containing peptides in water buffalo horn extract. The contents of -SH and peptide in water buffalo horn samples were determined by Ellman method and peptide assay kit， respectively. And then the peptides in water buffalo horn extract and enriched fraction were identified and analyzed by nano LC-MS/MS system. RESULTS The enrichment factor of Fe3O4@PDA@Au nano-materials in the enrichment of thiol-containing peptides in water buffalo horn extract was calculated as 23.41. A total of 4114 peptides were identified in the water buffalo horn extract， including 11 peptides containing Cys residue; a total of 2254 peptides were identified in the enriched fraction， including 709 peptides containing Cys residue. After enrichment， the ratio of the number of thiol-containing peptide to the total peptide in water buffalo horn extract increased from 0.003 to 0.314. The results indicated that the nano-material had a high efficiency on the enrichment of thiol-containing peptides in water buffalo horn extract. CONCLUSION Fe3O4@PDA@Au nano-materials can be used for the enrichment of thiol-containing peptides in water buffalo horn， which is of great significance for the basic research of water buffalo horn.
Determination Method of Starch Content in Chinese Medicine Water Extract： Comparative and Improvement
PENGJing, ZHUHua-xu, LIBo, MAOXiang-jun
2020, 36(3): 401-407.
[Abstract](358) [PDF 453KB](330)
Abstract:
OBJECTIVE To establish a rapid and accurate method for determination of starch content in Chinese medicine water extract. METHODS In this study， the starch content assay kit method was optimized for the traditional Chinese medicine water extraction system and the methodology was investigated. Then the starch simulation solution was taken as the research object. The content measurement results of three methods including iodine colorimetry， enzymatic hydrolysis method and the optimized starch content assay kit method were compared. RESULTS The optimized starch content assay kit method has good reproducibility and stable test results. The precision tests showed a relative standard deviation (n=6) of 1.91%. The accuracy test relative standard deviations (n=6) of the starch simulated solution and the aqueous extract were 1.82%~4.32%. The average recovery sampling rate of starch sample were 99.18%~104.41%， and the relative standard deviations (n=9) were 2.33%~4.94%. The best method was the optimized starch content assay kit method. CONCLUSION It shows that the optimized starch content assay kit method has good precision and accuracy， can be used as a detection method for determination of starch content in Chinese medicine water extract.
The Design of Specific Primers of Pheretima aspergillum<\i> and the Identification of Adulterants
ZHANGQian-cheng, WENHong-mei, LIUNa, YUANQi, LIURui, CUIXiao-bing, LISong, CHENSheng-jun
2020, 36(3): 408-413.
[Abstract](1564) [PDF 695KB](348)
Abstract:
OBJECTIVE The specific primers were designed to identify Pheretima aspergillum and its adulterants. METHODS The DNA of Pheretima aspergillum and its adulterants were extracted and amplified using the COⅠsequences as primers. The amplification products were sequenced afterwards. By comparing the COⅠsequences of Pheretima aspergillum with its adulterants， the variation sites were found in Pheretima aspergillum and the specific primers were designed to identify Pheretima aspergillum and its adulterants. PCR reaction system was optimized. RESULTS 750 bp fragments could be detected in the products of other animal medicinal materials using the COⅠsequences as primers， which meant the primers were not specific enough to distinguish the Pheretima aspergillum from its adulterants. Based on the COⅠsequences of amplification products， the specific primers PA-f/PA-r were designed. Through the established PCR reaction system， 574 bp fragments were amplified from DNA templates of Pheretima aspergillum. All the adulterants had no bands. CONCLUSION The designed primers sequences could accurately and successfully distinguish the Pheretima aspergillum from its adulterants， providing an effective method for the identification of Pheretima aspergillum.
Quality Evaluation of Toosendan Fructus Based on Content Determination and Pharmacological Activity
SU Hang, XUE Qian-qian, FEI Cheng-hao, LI Wei-dong, YIN Fang-zhou
2020, 36(3): 414-418.
[Abstract](484) [PDF 770KB](337)
Abstract:
OBJECTIVE To compare the contents and color values of two representative components， toosendanin and isotoosendanin， in different batches of toosendan fructus， and to compare the effects of different batches of toosendan fructus extracts on the expression of DR5 and the apoptosis of NSCLC cells induced by TRAIL. METHEODS The contents of toosendanin and isotoosendanin in toosendan fructus were determined by UHPLC-QqQ-MS/MS. The analysis was performed on Poroshell 120 SB C18 (2.1 mm×100 mm，1.8 μm) column， mobile phase was 0.1% formic acid aqueous solution and acetonitrile with flow rate of 0.3 mL/min. The injection volume was 1 μL， and the column temperature was 35 ℃. Meanwhile， spectrophotometer was used to determine the color value of the samples. Flow cytometry analysis was used to detect apoptosis and the expressions of cell death receptor DR5 for TRAIL in non-small cell lung cancer cell line A549. RESULTS The contents of toosendanin and isotoosendanin in 10 batches of samples were 1.024~4.128 mg/g and 0.010~0.045 mg/g， respectively. The average of colour value (L*，a*，b*) was 68.56，5.27 and 25.12. The extracts from 10 batches of toosendan fructus could significantly increase the expression of DR5 and restore the apoptotic sensitivity of A549 cells towards TRAIL. CONCLUSION The content determination and color measurement， combined with pharmacological activity can be successfully applied to accurately evaluate the quality of toosendan fructus.
Research Progress of GProtein Coupled Receptor MrgprD and its Implications for the Research of Chinese Medicine
WANG Di-jun, JIN Xiang, TANG Zong-xiang, WANG Chang-ming
2020, 36(3): 419-423.
[Abstract](551) [PDF 486KB](375)
Abstract:
Traditional Chinese medicine has the advantages of small side effects and clear curative effect in relieving itching and analgesia， but the molecular biological mechanisms are not clear. MrgprD (Mas-related gene D) is a member of the G-protein coupled receptor and is mainly expressed on peripheral primary sensory neurons. The study revealed the mechanism of involvement of MrgprD in neuropathic pain and discussed the relationship between itch and pain. Based on our own research results， we reviewed the current research status of MrgprD in recent decades. This review could provide a reference for the understanding of the function of MrgprD and the treatment of related diseases. Additionally， MrgprD may be the targets of antipruritic and analgesic Chinese herb and provide a new idea for the research of Chinese medicine.
Research Progress on Anti-Tumor Effect of Ligustrazine in Digestive System Neoplasms
LIHua, BIANYao-yao, YANGLi-li, ZHOUXi, ZENGLi
2020, 36(3): 424-427.
[Abstract](456) [PDF 409KB](385)
Abstract:
Digestive system neoplasms remain a major threat to residents' health in our country， with the second highest morbidity and mortality in China. Therefore， there is an urgent need for developing safe and effective anti-tumor drugs. The anti-tumor effect of plant extracts has gradually become a research focus. A growing attention has been paid to anti-cancer effects of Ligustrazine， which is the effective component of Chuanxiong. We reviewed the anti-neoplastic effect of Ligustrazine from the aspects of proliferation， apoptosis， metastasis， invasion and reversion of chemotherapy resistance in neoplasms of digestive system， providing a theoretical basis and foundation for further exploring the anti-cancer mechanisms of Ligustrazine and promoting the clinical application.

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