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      Vol. 29 ， Issue 9 ， 2023

        Original Article

      • CHI Xin,CHENG Dan-ying,SUN Xiu,LIU Shun-ai,WANG Rong-bing,CHEN Qin,XING Hui-chun
        Vol. 29, Issue 9, Pages: 771-781(2023) DOI: 10.1007/s11655-023-3542-2
        Abstract：Objective:To analyze the efficacy of Biejiajian Pill (BJJP) on intestinal microbiota in patients with hepatitis B cirrhosis/liver fibrosis, and explore its relationship with liver fibrosis.Methods:This was a prospective, randomized double-blind controlled trial. Using the stratified block randomization method, 35 patients with hepatitis B liver cirrhosis/liver fibrosis were randomly assigned (1:1) to receive entecavir (0.5 mg/d) combined with BJJP (3 g/time, 3 times a day) or placebo (simulator as control, SC group, simulator 3 g/time, 3 times a day) for 48 weeks. Blood and stool samples were collected from patients at baseline and week 48 of treatment, respectively. Liver and renal functions as well as hematological indices were detected. Fecal samples were analyzed by 16S rDNA V3-V4 high-throughput sequencing, and intestinal microbiota changes in both groups before and after treatment were compared, and their correlations with liver fibrosis were analyzed.Results:Compared with the SC group, there was no significant difference in liver function, renal function and hematology indices in the BJJP group, however, the improvement rate of liver fibrosis was higher in the BJJP group (94.4% vs. 64.7%,P=0.041). Principal coordinate analysis (PCoA) based on weighted Unifrac distance showed significant differences in intestinal microbiota community diversity before and after BJJP treatment (P <0.01 and P=0.003), respectively. After 48 weeks' treatment, the abundance levels of beneficial bacteria (Bifidobacteria, Lactobacillus, Faecalibacterium and Blautia) increased, whereas the abundance levels of potential pathogenic bacteria, including Escherichia coli, Bacteroides, Ruminococcus, Parabacteroides and Prevotella decreased, among which Ruminococcus and Parabacteroides were significantly positively correlated with degree of liver fibrosis (r=0.34,P=0.04;r=0.38,P=0.02), respectively. The microbiota in the SC group did not change significantly throughout the whole process of treatment.Conclusion:BJJP had a certain regulatory effect on intestinal microbiota of patients with hepatitis B cirrhosis/liver fibrosis (ChiCTR1800016801).  
        Keywords：Biejiajian Pill;chronic hepatitis B;liver fibrosis;liver cirrhosis;intestinal microbiota;randomized controlled trial;Chinese medicine   
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        Published：2023-08-29
      • WANG Wen,HE Qiao,WANG Ming-qi,XU Jia-yue,JI Peng,ZHANG Rui,ZOU Kang,SUN Xin
        Vol. 29, Issue 9, Pages: 782-790(2023) DOI: 10.1007/s11655-023-3634-z
        Abstract：Objective:To assess whether the use of Tanreqing (TRQ) Injection could show improvements in time to extubation, intensive care unit (ICU) mortality, ventilator-associated events (VAEs) and infection-related ventilator associated complication (IVAC) among patients receiving mechanical ventilation (MV).Methods:A time-dependent cox-regression analysis was conducted using data from a well-established registry of healthcare-associated infections at ICUs in China. Patients receiving continuous MV for 3 days or more were included. A time-varying exposure definition was used for TRQ Injection, which were recorded on daily basis. The outcomes included time to extubation, ICU mortality, VAEs and IVAC. Time-dependent Cox models were used to compare the clinical outcomes between TRQ Injection and non-use, after controlling for the influence of comorbidities/conditions and other medications with both fixed and time-varying covariates. For the analyses of time to extubation and ICU mortality, Fine-Gray competing risk models were also used to measure competing risks and outcomes of interest.Results:Overall, 7,685 patients were included for the analyses of MV duration, and 7,273 patients for the analysis of ICU mortality. Compared to non-use, patients with TRQ Injection had a lower risk of ICU mortality [Hazards ratios (HR) 0.761, 95% CI, 0.581–0.997], and was associated with a higher hazard for time to extubation (HR 1.105, 95% CI, 1.005–1.216), suggesting a beneficial effect on shortened time to extubation. No significant differences were observed between TRQ Injection and non-use regarding VAEs (HR 1.057, 95% CI, 0.912–1.225) and IVAC (HR 1.177, 95% CI, 0.929–1.491). The effect estimates were robust when using alternative statistic models, applying alternative inclusion and exclusion criteria, and handling missing data by alternative approaches.Conclusion:Our findings suggested that the use of TRQ Injection might lower mortality and improve time to extubation among patients receiving MV, even after controlling for the factor that the use of TRQ changed over time.  
        Keywords：Tanreqing Injection;critical care;mechanical ventilation;mortality;ventilator-associated events   
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        Published：2023-08-29
      • YE Miao-yong,ZHAO Fan,MA Ke,YAO Li-juan,ZHOU Kang,MA Jian-xiong,LYU Bo-dong,XU Zeng-bao
        Vol. 29, Issue 9, Pages: 791-800(2023) DOI: 10.1007/s11655-022-3532-9
        Abstract：Objective:To verify the effect of Buyang Huanwu Decoction (BHD) in ameliorating erectile dysfunction (ED) after radical prostatectomy (RP).Methods:The composition of BHD was verified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) analysis. Bilateral cavernous nerve crush injury (BCNI) in rats was used to mimic the neurovascular injury occurring after RP. By the envelope method, forty rats were randomly divided into 4 groups as follows: sham (cavernous nerves exposed only), model (BCNI), low-dosage BHD [LBHD, 12.8 g/(kg•d)], and high-dosage BHD [HBHD, 51.2 g/(kg•d)] groups, 10 rats in each group, feeding for 3 weeks respectively. Erectile function was evaluated by measuring intracavernosal pressure (ICP). Changes in the histopathology of corpus cavernosum (CC) were examined by hematoxylin-eosin staining. Meanwhile, the fibrosis of CC was measured by Masson's trichrome staining and Western blot was used to detect the expressions of collagen Ⅰ, transforming growth factor beta 1 (TGF-β1) and α-smooth muscle actin (α-SMA). Apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and Western blot for determining the expressions of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax). The oxidative stress in the CC were assessed by the superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) levels. The proteins expression of c-Jun N-terminal kinase (JNK) and c-Jun were detected by Western blot. In addition, the expression of α-SMA and p-c-Jun in the CC was observed by double immunofluorescence staining.Results:The UPLC-QTOF-MS/MS analysis showed that BHD contained calycosin-7-O-β-D-glucoside, ononin, calycosin and formononetin. Compared with the model group, LBHD and HBHD treatment improved the ICP and the circumference, area, and weight of CC (P<0.05 or P<0.01). Furthermore, LBHD and HBHD treatments increased CC smooth muscle content and decreased apoptosis index (P <0.05 or P <0.01). LBHD and HBHD also elevated SOD and expression level of α-SMA and Bcl-2, and reduced MDA and ROS levels, as well as expression of TGF-β1, collagen Ⅰ, Bax, p-c-JNK, p-JNK in the CC compared with the model group (P <0.05 or P <0.01). The double immunofluorescence staining showed that the fluorescence degree of p-c-Jun in both LBHD and HBHD treatment groups was significantly reduced, whereas the α-SMA expression increased (P <0.05 or P <0.01).Conclusions:BHD can improve ED of rats with BCNI, which is related to inhibiting fibrosis, apoptosis, and oxidative stress of CC. The ROS/JNK/c-Jun signaling pathway may play an important role in the process.  
        Keywords：Buyang Huanwu Decoction;erectile dysfunction;cavernous nerve injury;Chinese medicine;Jun N-terminal kinase   
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        Published：2023-08-29
      • LIU Hong,CHEN Wei-dong,HU Yang-lin,YANG Wen-qiang,HU Tao-tao,WANG Huan-lan,ZHANG Yan-min
        Vol. 29, Issue 9, Pages: 801-808(2023) DOI: 10.1007/s11655-022-3540-9
        Abstract：Objective:To investigate the effect of emodin on high glucose (HG)-induced podocyte apoptosis and whether the potential anti-apoptotic mechanism of emodin is related to induction of adenosine-monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)-mediated autophagy in podocytes (MPC5 cells) in vitro.Methods:MPC5 cells were treated with different concentrations of HG (2.5, 5, 10, 20, 40, 80 and 160 mmol/L), emodin (2, 4, 8 μmol/L), or HG (40 mmol/L) and emodin (4 μmol/L) with or without rapamycin (Rap, 100 nmol/L) and compound C (10 μmol/L). The viability and apoptosis of MPC5 cells were detected using cell counting kit-8 (CCK-8) assay and flow cytometry analysis, respectively. The expression levels of cleaved caspase-3, autophagy marker light chain 3 (LC3) Ⅰ/Ⅱ, and AMPK/mTOR signaling pathway-related proteins were determined by Western blot. The changes of morphology and RFP-LC3 fluorescence were observed under microscopy.Results:HG at 20, 40, 80 and 160 mmol/L dose-dependently induced cell apoptosis in MPC5 cells, whereas emodin (4 μmol/L) significantly ameliorated HG-induced cell apoptosis and caspase-3 cleavage (P <0.01). Emodin (4 μmol/L) significantly increased LC3-Ⅱ protein expression levels and induced RFP-LC3-containing punctate structures in MPC5 cells (P <0.01). Furthermore, the protective effects of emodin were mimicked by rapamycin (100 nmol/L). Moreover, emodin increased the phosphorylation of AMPK and suppressed the phosphorylation of mTOR. The AMPK inhibitor compound C (10 μmol/L) reversed emodin-induced autophagy activation.Conclusion:Emodin ameliorated HG-induced apoptosis of MPC5 cells in vitro that involved induction of autophagy through the AMPK/mTOR signaling pathway, which might provide a potential therapeutic option for diabetic nephropathy.  
        Keywords：emodin;diabetic nephropathy;autophagy;podocyte apoptosis;adenosine-monophosphate-activated protein kinase/mammalian target of rapamycin signaling pathways   
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        Published：2023-08-29
      • WEN Yong,ZHAN Yu,TANG Shi-yu,LIU Fang,WANG Qiu-xiao,KONG Peng-fei,TANG Xue-gui
        Vol. 29, Issue 9, Pages: 809-817(2023) DOI: 10.1007/s11655-022-3539-2
        Abstract：Objective:To explore the possible effects and mechanism of Zhizhu Decoction (ZZD) on the pathophysiology of slow transit constipation (STC).Methods:A total of 54 C57BL/6 mice was randomly divided into the following 6 groups by a random number table, including control, STC model (model), positive control, and low-, medium- and high-doses ZZD treatment groups (5, 10, 20 g/kg, namely L, M-, and H-ZZD, respectively), 9 mice in each group. Following 2-week treatment, intestinal transport rate (ITR) and fecal water content were determined, and blood and colon tissue samples were collected. Hematoxylin-eosin and periodic acid-Schiff staining were performed to evaluate the morphology of colon tissues and calculate the number of goblet cells. To determine intestinal permeability, serum levels of lipopolysaccharide (LPS), low-density lipoprotein (LDL) and mannose were measured using enzyme-linked immunosorbent assay (ELISA). Western blot analysis was carried out to detect the expression levels of intestinal tight junction proteins zona-occludens-1 (ZO-1), claudin-1, occludin and recombinant mucin 2 (MUC2). The mRNA expression levels of inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-4, IL-10 and IL-22 were determined using reverse transcription-quantitative reverse transcription reaction. Colon indexes of oxidative stress were measured by ELISA, and protein expression levels of colon silent information regulator 1/forkhead box O transcription factor 1 (SIRT1/FoxO1) antioxidant signaling pathway were detected by Western blot.Results:Compared with the model group, ITR and fecal moisture were significantly enhanced in STC mice in the M-ZZD and H-ZZD groups (P <0.01). Additionally, ZZD treatment notably increased the thickness of mucosal and muscular tissue, elevated the number of goblet cells in the colon of STC mice, reduced the secretion levels of LPS, LDL and mannose, and upregulated ZO-1, claudin-1, occludin and MUC2 expressions in the colon in a dose-dependent manner, compared with the model group (P <0.05 or P <0.01). In addition, ZZD significantly attenuated intestinal inflammation and oxidative stress and activated the SIRT1/FoxO1 signaling pathway (P <0.05 or P <0.01).Conclusion:ZZD exhibited beneficial effects on the intestinal system of STC mice and alleviated intestinal inflammation and oxidative stress via activating SIRT1/FoxO1 antioxidant signaling pathway in the colon.  
        Keywords：Zhizhu Decoction;intestinal barrier;slow transit constipation;oxidative stress;silent information regulator 1/forkhead box O transcription factor 1 signaling pathway   
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        Published：2023-08-29
      • Sahar Idris,Anuradha Mishra,Mohammad Khushtar
        Vol. 29, Issue 9, Pages: 818-824(2023) DOI: 10.1007/s11655-023-3734-9
        Abstract：Objective:To quantify phytochemicals using liquid chromatography and mass spectroscopy (LCMS) analysis and explore the therapeutic effect of Aesculus hippocastanum L. (AH) seeds ethanolic extract against gastric ulcers in rats.Methods:Preliminary phytochemical testing and LCMS analysis were performed according to standard methods. For treatment, the animals were divided into 7 groups including normal control, ulcer control, self-healing, AH seeds low and high doses, ranitidine and per se groups. Rats were orally administered with 10 mg/kg of indomethacin, excluding the normal control group (which received 1% carboxy methyl cellulose) and the per se group (received 200 mg/kg AH seeds extract). The test group rats were then given 2 doses of AH seeds extract (100 and 200 mg/kg, respectively), while the standard group was given ranitidine (50 mg/kg). On the 11th day, rats in all groups were sacrificed, and their stomach was isolated to calculate the ulcer index, and other parameters such as blood prostaglandin (PGE2), tissue superoxide dismutase (SOD), catalase (CAT), malonyldialdehyde (MDA), and glutathione (GSH). All isolated stomach tissues were analyzed for histopathological findings.Results:The phytochemical examination shows that the AH seeds contain alkaloids, flavonoids, saponins, phenolic components, and glycosides. LCMS analysis confirms the presence of quercetin and rutin. The AH seeds extract showed significant improvement in gastric mucosa conditions after indomethacin-induced gastric lesions (P <0.01). Further marked improvement in blood PGE2 and antioxidant enzymes, SOD, CAT, MDA and GSH, were observed compared with self-healing and untreated ulcer-induced groups (P <0.01). Histopathology results confirmed that AH seeds extract improved the mucosal layer and gastric epithelial membrane in treated groups compared to untreated ulcer-induced groups.Conclusions:LCMS report confirms the presence of quercetin and rutin in AH seeds ethanolic extract. The therapeutic effect of AH seeds extract against indomethacin-induced ulcer in rat model indicated the regenerated membrane integrity, with improved cellular functions and mucus thickness. Further, improved antioxidant enzyme level would help to reduce PGE2 biosynthesis.  
        Keywords：Aesculus hippocastanum L.;pharmacological action;phytochemical;physicochemical analysis;liquid chromatography and mass spectroscopy analysis;gastro-protective activity   
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        Published：2023-08-29
      • WANG Xi-xin,ZOU Hong-yuan,CAO Yong-na,ZHANG Xuan-ming,SUN Meng,TU Peng-fei,LIU Ke-chun,ZHANG Yun
        Vol. 29, Issue 9, Pages: 825-831(2023) DOI: 10.1007/s11655-022-3543-6
        Abstract：Objective:To investigate the anti-inflammatory activity of Radix Panacis quinquefolii extract (RPQE) and its therapeutic effects on inflammatory bowel disease (IBD).Methods:The 72-hour post-fertilization zebrafish was used to generate the local and systematic inflammation models through tail-amputation and lipopolysaccharide (LPS)-induction (100 μg/mL), respectively. The Tg(zlyz:EGFP) zebrafish was induced with 75 μg/mL 2,4,6-trinitrobenzene sulfonic acid (TNBS) for establishing the IBD model. The tail-amputated, LPS-, and TNBS-induced models were subjected to RPQE (ethanol fraction, 10–20 μg/mL) administration for 12 and 24 h, respectively. Anti-inflammatory activity of RPQE was evaluated by detecting migration and aggregation of leukocytes and expression of inflammation-related genes. Meanwhile, TNBS-induced fish were immersed in 0.2% (W/V) calcein for 1.5 h and RPQE for 12 h before photographing to analyze the intestinal efflux efficiency (IEE). Moreover, the expression of inflammation-related genes in these fish was detected by quantitative polymerase chain reaction.Results:Subject to RPQE administration, the migration and aggregation of leukocytes were significantly alleviated in 3 zebrafish models (P <0.01). Herein, RPQE ameliorated TNBS-induced IBD with respect to a significantly reduced number of leukocytes, improved IEE, and inhibited gene expression of pro-inflammatory factors (P <0.05 or P <0.01).Conclusion:RPQE exhibited therapeutic effects on IBD by inhibiting inflammation.  
        Keywords：Radix Panacis quinquefolii extract;anti-inflammatory;zebrafish;inflammatory bowel disease   
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        Published：2023-08-29

        Clinical Experience

      • WU Yu-he,XIAO Hai-yan,QUAN Ri-cheng,TANG Xu-dong,LIU Wei-yi,LYU Yan,CHEN Zhuo,LIU Chi,HU Xiao-mei
        Vol. 29, Issue 9, Pages: 832-837(2023) DOI: 10.1007/s11655-023-3603-6
        Abstract：Objective:To compare the clinical effect of arsenic-containing Qinghuang Powder (QHP) and low-intensity chemotherapy (LIC) in treatment of elderly acute myeloid leukemia (eAML) patients.Methods:Clinical data of 80 eAML patients treated at Xiyuan Hospital of China Academy of Chinese Medical Sciences from January 2015 to December 2020 were retrospectively analyzed. The treatment scheme was designed by real world study according to patients' preference, and patients were divided into a QHP group (35 cases) and a LIC group (45 cases). The median overall survival (mOS), 1-, 2-, and 3-year OS rates, and incidence of adverse events were compared between the two groups.Results:The mOS of 80 patients was 11 months, and the 1-, 2-, and 3-year OS rates were 45.51%, 17.96%, and 11.05%, respectively. The QHP and LIC groups demonstrated no significant difference in mOS (12 months vs. 10 months), 1- (48.57% vs. 39.65%), 2- (11.43% vs. 20.04%), and 3-year OS rates (5.71% vs. 13.27%, all P >0.05). Moreover, the related factors of mOS demonstrated no significant difference in patients with age >75 years (11 months vs. 8 months), secondary AML (11 months vs. 8 months), poor genetic prognosis (9 monthsvs. 7 months), Eastern Cooperative Oncology Group performance status score 3 (10 months vs. 7 months) and hematopoietic stem cell transplant comorbidity index 4 (11 months vs. 7 months) between the QHP and LIC groups (all P >0.05). However, the incidence of myelosuppression was significantly lower in the QHP group than that in the LIC group (28.57% vs. 73.33%, P <0.01).Conclusions:QHP and LIC had similar survival rates in eAML patients, but QHP had a lower myelosuppression incidence. Hence, QHP can be an alternative for eAML patients who do not tolerate LIC.  
        Keywords：acute myeloid leukemia;elderly;Qinghuang Powder;low-intensity chemotherapy;Chinese medicine   
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        Published：2023-08-29

        Data Mining Research

      • CHEN Hao-yu,LI Qi,ZHOU Ping-ping,YANG Tian-xiao,LIU Shao-wei,ZHANG Teng-fei,CUI Zhen,LYU Jing-jing,WANG Yan-gang
        Vol. 29, Issue 9, Pages: 838-846(2023) DOI: 10.1007/s11655-022-3538-3
        Abstract：Objective:To identify specific Chinese medicines (CMs) that may benefit patients with gastroesophageal reflux disease (GERD), and explore the action mechanism.Methods:Domestic and foreign literature on the treatment of GERD with CMs was searched and selected from China National Knowledge Infrastructure, China Science and Technology Journal Database, Wanfang Database, and PubMed from October 1, 2011 to October 1, 2021. Data from all eligible articles were extracted to establish the database of CMs for GERD. Apriori algorithm of data mining techniques was used to analyze the rules of herbs selection and core Chinese medicine formulas were identified. A system pharmacology approach was used to explore the action mechanism of these medicines.Results:A total of 278 prescriptions for GERD were analyzed, including 192 CMs. Results of Apriori algorithm indicated that Evodiae Fructus and Coptidis Rhizoma were the highest confidence combination. A total of 32 active ingredients and 66 targets were screened for the treatment of GERD. Enrichment analysis showed that the mechanisms of action mainly involved pathways in cancer, fluid shear stress and atherosclerosis, advanced glycation end product (AGE), the receptor for AGE signaling pathway in diabetic complications, bladder cancer, and rheumatoid arthritis.Conclusion:Evodiae Fructus and Coptidis Rhizoma are the core drugs in the treatment of GERD and the potential mechanism of action of these medicines includes potential target and pathways.  
        Keywords：gastroesophageal reflux disease;Chinese medicine;systematic pharmacology;data mining   
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        Published：2023-08-29

        Review

      • LI Han,YE Xiao-feng,SU Yang-shuai,HE Wei,ZHANG Jian-bin,ZHANG Qi,ZHAN Li-bin,JING Xiang-hong
        Vol. 29, Issue 9, Pages: 847-857(2023) DOI: 10.1007/s11655-022-3531-x
        Abstract：The latest guideline about ulcerative colitis (UC) clinical practice stresses that mucosal healing, rather than anti-inflammation, is the main target in UC clinical management. Current mucosal dysfunction mainly closely relates to the endoscopic intestinal wall (mechanical barrier) injury with the imbalance between intestinal epithelial cells (IECs) regeneration and death, as well as tight junction (TJ) dysfunction. It is suggested that biological barrier (gut microbiota), chemical barrier (mucus protein layer, MUC) and immune barrier (immune cells) all take part in the imbalance, leading to mechanical barrier injury. Lots of experimental studies reported that acupuncture and moxibustion on UC recovery by adjusting the gut microbiota, MUC and immune cells on multiple targets and pathways, which contributes to the balance of IEC regeneration and death, as well as TJ structure recovery in animals. Moreover, the validity and superiority of acupuncture and moxibustion were also demonstrated in clinic. This paper aims to review the achievements of acupuncture and moxibustion on mucosal healing and analyse the underlying mechanisms.  
        Keywords：acupuncture and moxibustion;ulcerative colitis;mucosal healing;intestinal epithelial cells regeneration and death;tight junction function;gut microbiota;review   
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        Published：2023-08-29
      • LU Chun-miao,YANG Yu-song,LU Yan-min,ZHU Ya-ping,ZHANG Ao,LYU Shi-chao,ZHANG Jun-ping
        Vol. 29, Issue 9, Pages: 857-864(2023) DOI: 10.1007/s11655-022-3288-2
        Abstract：Qishen Yiqi Dripping Pills (QSYQ) is a compound of Chinese medicine, which has been used to treat coronary heart disease and cardiac dysfunction. Its natural components include astragaloside Ⅳ, flavonoids, danshensu, protocatechualdehyde, salvianolic acid B, salvianolic acid A, ginsenosides Rg1, ginsenosides Rb1, and essential oils, etc. It exerts effects of nourishing qi and promoting blood circulation to relieve pain. In this review, the bioactive components of QSYQ and its effects for treating cardiovascular diseases and possible mechanism were summarized, providing references for further study and clinical application of QSYQ.  
        Keywords：Qishen Yiqi Dripping Pills;active ingredients;cardiovascular diseases;mechanisms;review   
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