Latest Issue
      2019 Year 25 Vol. 6 Issue

        Original Article

      • Pan ZHAO,Jun-bin LIANG,Zhong-yang DENG,Ming-jing WANG,Jia-yue QIN,Chong-jian CHEN,Xiao-mei HU
        2019, 25(6): 409-415. DOI: 10.1007/s11655-018-2977-3
        Abstract：Objective:To investigate the relationship between gene mutations and response to Compound Qinghuang Powder (复方青黄散, CQHP) in patients with myelodysplastic syndrome (MDS).Methods:Forty-three MDS patients after treatment with CQHP for 6 months were genotyped by ultra-deep targeted sequencing and the clinical data of patients were collected and the relationship between them was analyzed.Results:Up to 41.86% of patients harbored genet mutations, in most cases with more than one mutation. The most common mutations were in SF3B1, U2AF1, ASXL1, and DNMT3A. After treatment with CQHP, about 88.00% of patients no longer required blood transfusion, or needed half of prior transfusions.Conlusion:CQHP is an effective treatment for patients with MDS, especially those with gene mutations in SF3B1, DNMT3A, U2AF1, and/or ASXL1.  
        Keywords：myelodysplastic syndromes;arsenic;next-generation sequencing;realgar   
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        Published：2021-08-27
      • Wen-ting HE,Yan-hua ZHU,Tong ZHANG,Patima ABULIMITI,Fan-ye ZENG,Li-ping ZHANG,Ling-juan LUO,Xin-mei XIE,Hong-liang ZHANG
        2019, 25(6): 416-424. DOI: 10.1007/s11655-018-2997-z
        Abstract：Objective:To investigate the potential mechanisms that curcumin reverses 5-fluorouracil (5-FU) multidrug resistance (MDR).Methods:Cell growth and the inhibitory rate of curcumin (2–25 μg/mL) and/or 5-FU (0.05–1000 μg/mL) on human colon cancer HCT-8 and HCT-8/5-FU (5-FU-resistant cell line) were determined using cell counting kit-8 (CCK-8) assay. Apoptosis and cell cycle after 5-FU and/or curcumin treatment were detected by flow cytometry (FCM) and transmission electron microscopy (TEM). The expression of the multidrug resistance related factors p-glycoprotein (P-gp) and heat shock protein 27 (HSP-27) genes and proteins were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting (WB), respectively.Results:The inhibitory rate of curcumin or 5-FU on HCT-8 and HCT-8/5-FU cells proliferation at exponential phase were in a dosedependent manner, HCT-8 cell line was more sensitive to curcumin or 5-FU when compared the inhibitory rate of HCT-8/5-FU. The 50% inhibitory concentration (IC50) of combination 5-FU and curcumin (4.0 μg/mL) in HCT-8/5-FU was calculated as 179.26 μg/mL, with reversal fold of 1.85. Another IC50 of combination 5-FU and curcumin (5.5 μg/mL) in HCT-8/5-FU was calculated as 89.25 μg/mL, with reversal fold of 3.71. Synergistic effect of 5-FU and curcumin on HCT-8 and HCT-8/5-FU cells were found. The cell cycle analysis performed by FCM showed that HCT-8 and HCT-8/5-FU cells mostly accumulated at G0/G1 phase, which suggested a synergistic effect of curcumin and 5-FU to induce apoptosis. FCM analysis found that the percentage of apoptosis of cells treated with curcumin, 5-FU and their combination were significantly increased compared to the control group (P<0.05), and the percentage of apoptosis of the combination groups were slightly higher than other groups (P<0.05). The mRNA levels of P-gp (0.28±0.02) and HSP-27 (0.28±0.09) in HCT-8/5-FU cells treated with combination drugs were lower than cells treated with 5-FU alone (P-gp, 0.48±0.07, P=0.009; HSP-27, 0.57±0.10, P=0.007). The protein levels of P-gp (0.25±0.06) and HSP-27 (0.09±0.02) in HCT-8/5-FU cells treated with combination drugs were decreased when compared to 5-FU alone (P-gp, 0.46±0.02, P=0.005; HSP-27, 0.43±0.01, P=0.000).Conclusions:Curcumin can inhibit the proliferation of human colon cancer cells. Curcumin has the ability of reversal effects on the multidrug resistance of human colon cancer cells lines HCT-8/5-FU. Down-regulation of P-gp and HSP-27 may be the mechanism of curcumin reversing the drug resistance of HCT-8/5-FU to 5-FU.  
        Keywords：curcumin;Chinese medicine;5-fluorouracil;multidrug resistance;HCT-8;HCT-8/5-FU;colon cancer;P-gp;HSP-27   
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        Published：2021-08-27
      • Hao-jiang ZUO,Shan LIU,Chun YAN,Li-man LI,Xiao-fang PEI
        2019, 25(6): 425-430. DOI: 10.1007/s11655-018-2983-5
        Abstract：Objective:To examine the effect of the aqueous extract of Ligustrum robustum on tumor growth in vitro and in vivo and explore the possible molecular mechanisms.Methods:In in vitro study, cell viabilities of human cervical carcinoma cells (HeLa), human breast cancer cells (MCF-7), human prostate cancer cells (PC-3), human hepatoma cells (7721) and human colon carcinoma cells (SW480) were evaluated with cell counting kit-8. For L. robustum-treated Hela cells, early or late apoptosis were evaluated by annexin V/PI staining. Mitochondrial membrane potential was measured by staining cells with JC-1. Apoptosis was monitored by nuclear morphology based on chromatin condensation and fragmentation by 4',6-diamidino-2-phenylinole (DAPI) staining. Caspase-3 and -8 activity levels were measured by a colorimetric assay. In vivo, to evaluate the possible mechanism of L. robustum-mediated antitumor effect, nude mouse xenograft study was also conducted.Results:In in vitro study, L. robustum was found to be toxic to HeLa, MCF-7, PC-3, 7721, SW480, with an half maximal inhibitory concentration value of 2–5 mg/mL (P<0.05). Moreover, externalization of phosphatidylserine, loss of mitochondrial membrane potential, DNA fragmentation and activation of caspase-3 and -8 were detected in L. robustum-treated Hela cells. Using a nude mouse model bearing Hela xenografts, we found that L. robustum reduced tumor volume and tumor weight (P<0.05), but had no effect on body weight and histological damage of important organs. Intraperitoneal injection of L. robustum caused a significant reduction in serum aspartate transaminase and alanine transaminase levels (P<0.05). Furthermore, cleaved caspase-3-positive and terminal nucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells were observed in L. robustum-treated tumor tissues.Conclusions:L. robustum inhibits tumor cell growth both in vitro and in vivo by inducing apoptosis in a caspasedependent way without apparent hepatic toxicity and histological damage, which may offer partial scientific support for the ethnopharmacological claims of L. robustum as a herbal tea for its antitumor activity.  
        Keywords：Ligustrum robustum;antitumor effect;Ku-Ding-Cha;herbal tea   
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        Published：2021-08-27
      • Shi-hua LUO,Song-lin SONG,Chuan-sheng ZHENG,Wei-yong LI,Yong WANG,Xiang-wen XIA,Gan-sheng FENG
        2019, 25(6): 431-438. DOI: 10.1007/s11655-017-2953-3
        Abstract：Objective:To evaluate the characteristics of Bletilla striata microspheres (BSMs) and its effects as an embolic agent in a rabbit model.Methods:BSMs were prepared with an emulsification-cool condensation-chemical cross-linking method. The characteristics of BSMs in vitro were observed. Embolization experiments were performed in renal artery of rabbit and in a rabbit liver VX2 carcinoma model. Seventy-two New Zealand rabbits were divided into 2 groups, and the right renal artery was embolized with BSMs (200 μm in diameter) in the experimental group and with polyvinyl alcohol (PVA) of the same size in the control group. The pathological findings were examined with hematoxylin-eosin and Masson stainings. Liver and renal functions were tested before and after embolization. VX2 tumor was transplanted in 15 New Zealand rabbits, which were randomly divided into 3 groups (n=5). Group A were treated with saline, group B with a mixture of doxorubicin and lipiodol, and group C with hepatic arterial infusion of BSMs (200 μm in diameter). Tumor growth rate was evaluated by magnetic resonance imaging scan. Apoptosis-related factors (bax, bcl-2) and tumor vascular endothelial cell growth factor (VEGF) were evaluated through immunohistochemical staining.Results:The characteristics of BSMs in vitro were in full compliance with the requirements for use in interventional procedures. In the renal artery embolization experiment, after BSMs intervention, it was more difficult to form collateral circulation than that with PVAs, and the kidney manifested atrophy and calcification. There were no significant difference of liver and renal functions in rabbits between groups. In the liver VX2 carcinoma embolization experiment, compared with group A, the growth rate of VX2 liver tumor and Bcl-2 levels was reduced, while apoptosis index, Bax, and VEGF were increased in group B (P<0.05). There were no significant difference between groups B and C (P>0.05).Conclusions:The characteristics of BSMs in vitro and in vivo meet the requirements for its use as an embolic agent in interventional approaches.  
        Keywords：Bletilla striata;Chinese medicine;microsphere;characteristic;embolization   
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        Published：2021-08-27
      • Shan LIN,Jiu-mao LIN,Ling ZHANG,Da-xin CHEN,Fei XIAO,Hong-wei CHEN,You-qin CHEN,Yu-ling ZHU,Jian-feng CHU,Jun PENG
        2019, 25(6): 439-445. DOI: 10.1007/s11655-018-2976-4
        Abstract：Objectives:To investigate the protective effects of Shexiang Tongxin Dropping Pill (麝香通心滴丸, STP) on Na2S2O4-induced hypoxia-reoxygenation injury in cardiomyoblast H9c2 cells.Methods:The cell viability and levels of mRNA and protein expression in H9c2 cells were determined following Na2S2O4-induced hypoxia using Hoechst staining, annexin V/propidium iodide (PI) flow cytometry, real-time polymerase chain reaction and Western blot analysis.Results:STP pretreatment significantly increased the viability and inhibited aberrant morphological changes in H9c2 cardiomyoblast cells induced by Na2S2O4 treatment (P<0.05). In addition, STP pretreatment attenuated Na2S2O4-induced hypoxic damage, down-regulated the expression of pro-apoptotic Bax, and up-regulated the expression of anti-apoptotic Bcl-2 in H9c2 cells (P<0.05).Conclusions:STP was strongly cardioprotective in hypoxia-reoxygenation injury by preventing hypoxic damage and inhibiting cellular apoptosis. These results further support the use of STP as an effective drug for the treatment of ischemic heart disease.  
        Keywords：myocardial ischemia;apoptosis;Shexiang Tongxin Dropping Pill;Chinese medicine;Na2S2O4-induced hypoxia-reoxygenation injury   
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        Published：2021-08-27
      • Yan-min LIU,Liu-luan ZHU,Rui LI,Jin-liang ZHANG,Shan-shan YAO,Xiao-bing ZHOU,Hui ZENG,Xian-bo WANG
        2019, 25(6): 446-453. DOI: 10.1007/s11655-015-2141-2
        Abstract：Objective:To investigate the hepatoprotective effect of Xijiao Dihuang Decoction (犀角地黄汤, XJDHD) on lipopolysaccharide (LPS)- and tumor necrosis factor alpha (TNF-α)-induced acute liver failure (ALF) as well as the underlying mechanism of action, and to clarify the key herbs and components of XJDHD.Methods:LPS/D-galactosamine (D-GalN) or TNF-α/D-GalN were intraperitoneally injected into C57BL/6J mice to induce ALF. Simultaneously, XJDHD or its individual herbs and components were orally administered. Survival rates, transaminase levels in serum, and hepatic histology were examined to evaluate the effects of XJDHD. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and real-time polymerase chain reaction were additionally performed to expound the mechanism underlying the anti-apoptotic activity of XJDHD.Results:Oral administration of XJDHD protected mice from lethal liver failure induced by LPS and TNF-α, with notable amelioration of liver injury in histology and a significant decrease in transaminase levels in serum. XJDHD significantly inhibited apoptosis of hepatocytes and enhanced expression of the antiapoptosis genes, c-Flip, Iap1, Gadd45b and A20 (all P<0.05). In addition, Rehmannia glutinosa Libosch. was identified as the key herb of XJDHD and galactose as the effective component of Rehmannia glutinosa Libosch. that protects against ALF.Conclusions:XJDHD inhibits TNF-α-induced apoptosis of hepatocytes by promoting the expression of nuclear factor κB-regulated anti-apoptotic genes. Rehmannia glutinosa Libosch. may be the most effective herb of XJDHD and galactose is an active component in this protection.  
        Keywords：Xijiao Dihuang Decoction;Chinese medicine;Rehmannia glutinosa Libosch.;acute liver failure;lipopolysaccharides;tumor necrosis factor alpha   
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        Published：2021-08-27
      • Xuan YU,Xin-pei WANG,Xiao-jin YAN,Jing-fei JIANG,Fan LEI,Dong-ming XING,Yue-ying GUO,Li-jun DU
        2019, 25(6): 454-461. DOI: 10.1007/s11655-017-2952-4
        Abstract：Objective:To explore the anti-nociceptive effect of patchouli alcohol (PA), the essential oil isolated from Pogostemon cablin (Blanco) Bent, and determine the mechanism in molecular levels.Methods:The acetic acid-induced writhing test and formalin-induced plantar injection test in mice were employed to confirm the effect in vivo. Intracellular calcium ion was imaged to verify PA on mu-opioid receptor (MOR). Cyclooxygenase 2 (COX2) and MOR of mouse brain were expressed for determination of PA's target. Cellular experiments were carried out to find out COX2 and MOR expression induced by PA.Results:PA significantly reduced latency period of visceral pain and writhing induced by acetic acid saline solution (P<0.01) and allodynia after intra-plantar formalin (P<0.01) in mice. PA also up-regulated COX2 mRNA and protein (P<0.05) with a down-regulation of MOR (P<0.05) both in in vivo and in vitro experiments, which devote to the analgesic effect of PA. A decrease in the intracellular calcium level (P<0.05) induced by PA may play an important role in its anti-nociceptive effect. PA showed the characters of enhancing the MOR expression and reducing the intracellular calcium ion similar to opioid effect.Conclusions:Both COX2 and MOR are involved in the mechanism of PA's anti-nociceptive effect, and the up-regulation of the receptor expression and the inhibition of intracellular calcium are a new perspective to PA's effect on MOR.  
        Keywords：Chinese medicine;patchouli alcohol;analgesic effect;mu-opioid receptor;cyclooxygenase 2   
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        Published：2021-08-27

        Acupuncture Research

      • Wen-sheng ZHAO,Zhen-ni JIANG,Hui SHI,Lu-lu XU,Yue YANG,Ying-chao WANG
        2019, 25(6): 462-467. DOI: 10.1007/s11655-018-3057-4
        Abstract：Objective:To study the effects of electroacupuncture (EA) in chronic constrictive injury (CCI) rat model and the expression of N-methyl-D-aspartate receptor type 2B (NR2B) in ipsilateral spinal dorsal horn in rats to explore the analgesic mechanisms of EA.Methods:According to the random number table, totally 180 rats were evenly divided into a sham group, a CCI group, and an EA group. CCI model was conducted with four 4–0 chromic gut ligatures loosely ligated around the left sciatic nerve 1 cm above the trifurcation. Rats in the EA group received 2 Hz EA therapy bilaterally at acupoints of Zusanli (ST 36) and Sanyinjiao (SP 6) once daily (30 min/d) for 30 days after surgery. Paw withdrawal thresholds (PWTs) were measured on 0 (baseline), 1, 3, 7, 15, 30 days after surgery. Rats were sacrificed on 0, 1, 3, 7, 15 and 30 days after surgery, and the L4–5 segments of spinal cord were removed to detect the expression of NR2B by immunohistochemistry and quantitative polymerase chain reaction.Results:PWTs in the CCI group were significantly lower than the sham group at Day 1–30 after surgery, and reached its lowest at Day 1 (P<0.01). After EA treatment, the PWTs recovered rapidly and were significantly higher than those in the CCI group on 3, 7, 15 and 30 days after surgery (P<0.01). The numbers of NR2B-immunoreactive cells of the CCI group significantly increased after CCI surgery compared with the sham group (P<0.01). Compared with the CCI group, stimulation of EA markedly decreased the numbers of NR2B-immunoreactive cells at Day 3, 7, 15 and 30 (P<0.05). In the sham group, NR2B mRNA was expressed at a low level. It increased after CCI surgery, which increased rapidly at Day 7 (P<0.01) and reached its peak value at Day 15 (P<0.01). After EA stimulation, relative quantity of NR2B mRNA expression was less than that in the CCI group at Day 15 and 30 (P<0.05).Conclusions:Low frequency of EA had antinociceptive effect in CCI rat model. The analgesic effects of EA might be through the inhibition of NR2B.  
        Keywords：sciatica;electroacupuncture;chronic constrictive injury;spinal dorsal horn;N-methyl-D-aspartate receptor type 2B;Chinese medicine   
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        Published：2021-08-27

        Evidence-Based Integrative Medicine

      • Jun ZHAO,Xing LIAO,Hui ZHAO,Zhi-geng LI,Nan-yue WANG,Li-min WANG
        2019, 25(6): 471-480. DOI: 10.1007/s11655-018-2982-6
        Abstract：Objective:To evaluate the effectiveness and safety of Chinese herbs in the treatment of sub-health systematically.Methods:Nine databases were systematically and extensively searched to collect randomized controlled trials (RCTs) about Chinese herbs in the treatment of sub-health. The outcomes included overall effective rate, main symptoms, quality of life, etc. Literature screening, data extraction and quality assessment were conducted according to Cochrane Handbook 5.1. Meta-analysis was conducted to the included literature with Review Manager Software.Results:Seventy-two studies involving 9,296 patients with sub-health were included with 4,908 patients in experimental groups and 4,387 patients in control groups. The overall quality of included clinical research was not high. In the aspect of improving overall effective rate, relieving main symptoms, Pittsburgh sleep quality index (PSQI) score, Athens Insomnia Scale (AIS) score, Fatigue Scale-14 (FS-14), Cornell Medical Index (CMI) score and discontinuation rate, the effects of experimental groups were better than that of control groups. According to available research reports, adverse reactions in Chinese herb groups were mainly mild gastrointestinal symptoms, which did not affect the treatment.Conclusion:Chinese herbs have a curative effect in the treatment of sub-health. However, there are no clear criteria for diagnosis and curative effectiveness judgment globally, which would affect the accuracy of curative effect evaluation.  
        Keywords：sub-health;randomized controlled trials;Chinese medicine;systematic review   
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        Published：2021-08-27
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