Chinese Journal of Integrative Medicine

Mechanisms of Huangqi Decoction Granules (黄芪汤颗粒剂) on Hepatitis B Cirrhosis Patients Based on RNA-Sequencing Abstract Objective To explore the action mechanisms of Huangqi Decoction Granules (黄芪汤颗粒剂, HQDG) on hepatitis B cirrhosis. Methods A total of 85 patients with hepatitis B cirrhosis were randomly divided into HQDG group (42 cases) and control group (43 cases) by a random number table and were treated with HQDG or placebo for 48 weeks (6 g per times and orally for 3 times a day), respectively. After RNA-sequencing of serum samples extracted from the patients, the differentially expressed genes (DEGs) in HQDG and control groups before and after treatment were separately screened. The DEGs were then performed pathway enrichment analysis and proteinprotein interaction (PPI) network analysis. The expression levels of key genes were detected by quantitative realtime polymerase chain reaction (qRT-PCR). Results After the investigation, 4 and 3 cases were respectively excluded from HQD and control groups because of the incomplete data. Additionally, 3 and 5 cases were lost to follow up in HQD and control groups respectively. Finally, a total of 70 cases with good compliance were included for further DEGs analysis. A total of 1,070 DEGs (including 455 up-regulated genes and 615 down-regulated genes) in HQDG group and 227 DEGs (including 164 up-regulated genes and 63 down-regulated genes) in the control group were identified after treatment. Compared with the control group, 1,043 DEGs were specific in HQDG group. Besides, 1 up-regulated transcription factor (TF, such as GLI family zinc finger 1, GLI1) and 25 down-regulated TFs (such as drosophila mothers against decapentaplegic proteinfamily member 2, SMAD2) were identified. Pathway enrichment analysis showed that down-regulated Ras homolog gene family member A (RHOA) was enriched in pathogenic Escherichia coli infection. In the PPI network, up-regulated epidermal growth factor receptor (EGFR), and down-regulated cell division cycle 42 (CDC42) as well as v-akt murine thymoma viral oncogene homolog 1 (AKT1) had higher degrees. Moreover, long non-coding RNAs (lncRNA) growth arrest-specific 5 (GAS5) was involved in the lncRNA-target regulatory network. Furthermore, qRT-PCR revealed that expression levels of CDC42 and GLI1 had significant differences in HQDG group before and after treatment (P<0.05). Conclusions CDC42 and GLI1 may be the targets of HQDG in patients with hepatitis B cirrhosis. Additionally, SMAD2, EGFR, AKT1, RHOA and GAS5 might be associated with the curative effect of HQDG on hepatitis B cirrhosis patients.

Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway Abstract Objective To investigate whether ginsenoside-Rb1 (Gs-Rb1) improves the CoCl-induced autophagy of cardiomyocytes via upregulation of adenosine 5′-monophosphate-activated protein kinase (AMPK) pathway. Methods Ventricles from 1- to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group (normal level oxygen), hypoxia group (500 μmol/L CoCl2), Gs-Rb1 group (200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2), Ara A group (500 μmol/L Ara A + 500 μmol/L CoCl2), Ara A+ Gs-Rb1 group (500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) + 500 μmol/L CoCl2], and AICAR+Gs-Rb1 group (1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2). Cells were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cardiac troponin I (cTnI) levels were detected by enzyme-linked immunosorbent assay (ELISA). AMPK activity was assessed by 2′,7′-dichlorofluorescein diacetate (DCFH-DA) ELISA assay. The protein expressions of Atg4B, Atg5, Atg6, Atg7, microtubule-associated protein 1A/1B-light chain 3 (LC3), P62, and active-cathepsin B were measured by Western blot. Results Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes (P<0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A (P<0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A (P<0.01) and was not affected by AICAR =0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B II, the LC3B II/I ratio and cathepsin B activity of hypoxia cardiomyocytes (P<0.05), each of these protein levels was significantly enhanced by Ara A (all P<0.01), but was not affected by AICAR (all P>0.05). Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes (P<0.05). The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A (P<0.05) and were not affected by AICAR (P=0.871). Conclusion Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.

Research Progress on Chinese Medicine Immunomodulatory Intervention for Chronic Primary Immune Thrombocytopenia: Targeting Cellular Immunity Abstract Chronic primary immune thrombocytopenia (CITP) is the most common acquired autoimmune disease that seriously threaten the physical and mental health of patients. Compared with Western medicine treatment, the intervention and treatment of Chinese medicine (CM) has shown certain therapeutic advantages. This paper reviewed the new pathogenesis progress on T cell immune abnormality in CITP, and CM studies on interferes effects of cellular immune regulation of CITP in recent years. Qi deficiency failing to control blood and internal obstruction of blood stasis are the two common types of CM syndromes in CITP patients, the corresponding treatments include invigorating Pi (Spleen), supplementing qi, activating blood, as well as tonifying qi and activating yang, regulating Gan (Liver) to invigorate Pi. The authors also mentioned the problems in the research field of CM for CTIP treatment, and put forward new ideas for the research in the future.

Acupoint Catgut Embedding Alleviates Insomnia in Different Chinese Medicine Syndrome Types: A Randomized Controlled Trial Abstract Objective To investigate the effects and safety of catgut embedding on alleviating insomnia. Methods Totally 510 patients with insomnia were divided into 5 Chinese medicine (CM) syndrome types: Xin (Heart) and Pi (Spleen) deficiency, yin deficiency with excess fire, Xin and gut qi deficiency, Wei (Stomach) disorder, and qi and blood deficiency, respectively. These 5 types of patients were randomly assigned to a catgut embedding group, an acupuncture group or a medication group (30 cases in Xin and Pi deficiency type, Wei disorder type, Xin and gut qi deficiency type, respectively; 40 cases in yin deficiency with excess fire type and qi and blood deficiency type, respectively). In the catgut embedding group, patients were treated by implanting catgut into acupoints once every 10 days for a total of 30 days. In the acupuncture group, patients were treated with acupuncture once per day over 30 days (excluding weekends); and patients in the medication group took 1 mg Eurodin Tablet orally every night for 30 days. Pittsburgh Sleep Quality Index (PSQI) was evaluated before treatment, on 30 and 60 days after the first treatment, respectively. The International Unified Sleep Efficiency Value (IUSEV) was measured at 30 and 60 days. The safety was evaluated after treatment and adverse events were analyzed. Results The objective PSQI scores including subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbance, daytime dysfunction, and total scores at 30 days were significantly improved compared with pre-treatment in the catgut embedding and acupuncture groups (P<0.01 or P<0.05). At 30 days, the PSQI scores in catgut embedding group were superior to the medication group in the patients with each type of insomnia, with the exception of sleep duration (P<0.01 or P<0.05). At 60 days, significant differences were found between the catgut embedding group and the medication group (P<0.01 for all indices). The IUSEV scores in the catgut embedding group were significantly higher than the acupuncture group at 60 days, and the scores in acupuncture group were higher than the medication group at 30 days (P<0.05 for all types). No severe adverse events were found in this study. Conclusions Acupoint catgut embedding and acupuncture were more effective than medication in alleviating insomnia syndrome in different Chinese medicine syndrome type. However, the sustained effects of acupoint catgut embedding were superior to acupuncture.

Wet-Cupping Is Effective on Persistent Nonspecific Low Back Pain: A Randomized Clinical Trial Abstract Objective To compare the possible effects of wet-cupping therapy with conventional therapy on persistent nonspecific low back pain (PNSLBP). Methods In this randomized clinical trial, 180 participants with the mean age of 45±10 years old, who had been suffering from PNSLBP were randomly assigned to wet-cupping and conventional groups. The wet-cupping group was treated with two separate sessions (4 weeks in total) on the inter-scapular and sacrum area. In the conventional group, patients were conservatively treated using rest (6 weeks) and oral medications (3 weeks). The primary and the secondary outcome were the quantity of disability using Oswestry Disability Index (ODI), and pain intensity using Visual Analogue Scale (VAS), respectively. Results There was no significant difference in demographic characteristics (age, gender, and body mass index) between the two groups (P>0.05). Therapeutic effect of wet-cupping therapy was comparable to conventional treatment in the 1st month follow-up visits (P<0.05). The functional outcomes of wet-cupping at the 3rd and 6th month visits were significantly increased compared to the conventional group. Final ODI scores in the wet-cupping and conventional groups were 16.7 ± 5.7 and 22.3 ± 4.5, respectively (P<0.01). Conclusion Wet-cupping may be a proper method to decrease PNSLBP without any conventional treatment. The therapeutic effects of wet-cupping can be longer lasting than conventional therapy. (Registration No. IRCT2013021672741))

Identification of Chinese Medicine Constitution in Public Health Services Abstract Public health is the science and art of preventing disease, prolonging life and prompting physical and mental health through evaluation, policy development and sanitary measures. The development of public health has become an important part of health care reform in China, and it requires the participation of Chinese medicine (CM) in the practice of public health. This study analyzed the problems of CM services in public health, suggested that the identification of CM constitution could propose an application paradigm of CM services in public health, and described from aspects of theory, technology and application. The identification of CM constitution gained national approval and support of government and industry. With the application and popularization of this technology, it will promote the establishment of characteristic CM preventive care system and the development of public health.

Impact of Hydroalcoholic Extract of Humulus Lupulus L. on Sperm Quality, Reproductive Organs and Hormones in Male Rats Abstract Objective To investigate the impact of Humulus Lupulus L. hydroalcoholic extract on the body weights, reproductive organs, sperm quality and hormone levels in male rats. Methods By simple random sampling method, seventy male Sprague-Dawley rats were randomly assigned to 7 groups including control group [distilled water, 1 mL/(kg•d)], Tween 80 group [25% Tween 80 solution, 1 mL/(kg•d)], olive oil group [olive oil, 1 mL/(kg•d)], diethyl stilbestrol (DES) group [DES, 100 μg/(kg•body weight)], H50, H150 and H450 [50, 150 and 400 mg/(kg•d) of Humulus Lupulus L extract, respectively]. The administration was performed via gavage once daily for 7 weeks. Body and reproductive organs weights including testes, seminal vesicles, epididymis and prostate were weighted and epididymal sperm quality were determined by digital balance. Blood samples were collected and serum free testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estrogen (E2) levels were measured by rat specific enzyme-linked immunosorbent assay. Results The percentage increase in mean body weights of rats in the DES and H50, H150 and H450 groups decreased significantly compared to olive oil and Tween 80 groups (all P<0.05). The weights of seminal vesicle, epididymis and testes in rats receiving H50 were significantly higher than the control group (P<0.05 or P<0.01). The sperm count in the rats receiving H50 was significantly lower than the control group (P<0.05). The sperm motile characteristics of the rats receiving hydroalcoholic extract at and DES were significantly lower than those of the control or rats receiving vehicles (all P<0.05). In H50, H150, H450 and DES groups, T and LH levels were decreased, and E2 was significantly increased compared to the control (P<0.05 or P<0.01). The FSH level did not change in all groups (P>0.05). Conclusion Humulus Lupulus L. extract significantly increased the seminal vesicle and testes weights and reduced the sperm motility.

Tetramethylpyrazine Improves Postoperative Tissue Adhesion: A Drug Repurposing Abstract Plants are known to possess plenty of pharmacological activities as a result of various phytoconstituents. Tetramethylpyrazine (TMP), one of the most widely used medicinal compound isolated from traditional Chinese herb, is usually employed for anti-oxidation, anti-inflammation, anti-platelet aggregation, anti-lipid, anti-fibrosis, as well as activating blood, removing stasis, dilating small arteries, improving microcirculation and antagonizing calcium. In the present paper, the anti-adhesion effect of TMP were reviewed. TMP was found to play a multi-target and muti-link role in anti-adhesion by inhibiting hyperplasia of collagen and overexpression of adhesion-related factors and reducing the concentration of white blood cells and fibrin in plasma. Because previous studies mostly focused on in vitro experiments and animal experiments, there is an urgent need for clinical research with abundant indicators to further prove its anti-adhesion potency. Future basic research should concentrate on the development of TMP as a biological material.

Diagnostic Accuracy of Chinese Medicine Diagnosis Scale of Phlegm and Blood Stasis Syndrome in Coronary Heart Disease: A Study Protocol Abstract Background Phlegm and blood stasis syndrome (PBSS) is one of the main syndromes in coronary heart disease (CHD). Syndromes of Chinese medicine (CM) are lack of quantitative and easy-implementation diagnosis standards. To quantify and standardize the diagnosis of PBSS, scales are usually applied. Objective To evaluate the diagnostic accuracy of CM diagnosis scale of PBSS in CHD. Methods Six hundred patients with stable angina pectoris of CHD, 300 in case group and 300 in control group, will be recruited from 5 hospitals across China. Diagnosis from 2 experts will be considered as the “gold standard”. The study design consists of 2 phases: pilot test is used to evaluate the reliability and validity, and diagnostic test is used to assess the diagnostic accuracy of the scale, including sensitivity, specificity, likelihood ratio and area under the receiver operator characteristic (ROC) curve. Discussion This study will evaluate the diagnostic accuracy of CM diagnosis scale of PBSS in CHD. The consensus of 2 experts may not be ideal as a “gold standard”, and itself still requires further study. (No. ChiCTR-OOC-15006599).

Therapeutic Effect of Ershen Pill (二神丸) Extract on Pi (Spleen)-Shen (Kidney) Yang Deficiency-Induced Diarrhea in Rat Model Abstract Objective To investigate whether Ershen Pill (ESP, 二神丸) could alleviate the symptom of Pi (Spleen)-Shen (Kidney) yang deficiency (PSYD)-induced diarrhea in rat model and explore its anti-diarrhea mechanism. Methods Seventy-five Sprague-Dawley rats were divided into 5 groups by a random number table, including control, positive, model, low-dose (LD) and high-dose (HD) ESP groups, 15 rats in each group. All the rats, except those in the control group, were developed PSYD induced-diarrhea based on its pathology and etiology. The rats in positive, LD and HD ESP groups were treated with Shenling Baizhu Pill (参苓白术丸), LD (1.05 g/kg) or HD (3.50 g/kg) ESP petroleum ether extract once a day for 2 weeks, respectively. Body weight change and diarrhea index were measured. The histology scores of the kidney were evaluated via hematoxylin and eosin (HE) staining. Aquaporin-3 (AQP3) expression in the colon was analyzed by immunofluorescence, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot, respectively. Results Compared with the model group, oral administration of LD and HD ESP prevented body weight loss and inhibited diarrhea after 2-week treatment (P<0.05). Kidney deterioration was impeded, and the histology score in LD and HD ESP groups were 8.2 and 10.5, respectively, which were both higher than those in the model group (P<0.05). In addition, ESP treatment alleviated rat colitis, and HD ESP significantly improved the AQP3 positive staining intensity in the colon tissue compared with the model group. The result from Western blot revealed that AQP3 protein synthesis in colon tissue of LD and HD ESP groups increased by 2.1- and 5.9-fold compared with the model group (P<0.05). qRT-PCR result showed that AQP3 gene expression in the HD ESP group was also up-regulated by 2.5-fold normalized to the model group (P<0.05). Conclusion ESP extract effectively alleviates the symptoms of PSYD and relieves PSYD-induced diarrhea by improving AQP3 synthesis in the colon.