Mechanic evaluation of Wu-Mei-Pill on colitis-associated colorectal cancer: An integrated transcriptomics, metabolomics, and experimental validation study.

BACKGROUND: Chronic intestinal inflammatory diseases play a crucial role in the onset of colorectal cancer (CRC). Effectively impeding the progression of colitis-associated colorectal cancer (CAC) can be instrumental in hindering CRC development. Wu-Mei-Pill (WMP), a formulation comprising various herbal extracts, is clinically employed for CAC treatment, yet the underlying mechanism of WMP's efficacy in CAC remains unclear. Our study firstly demonstrated the effects and mechanisms of WMP on transcriptional and metabolic levels based on integrated transcriptomics and untargeted metabolomics and relative experimental validations. MATERIALS AND METHODS: A CAC mouse model was established through a single injection of azoxymethane (AOM) followed by intermittent dextran sodium sulfate (DSS) intervention, with subsequent WMP administration. Initially, the therapeutic impact of WMP on the CAC model was assessed by observing survival rate, body weight change, colon length, tumor number, tumor load, and pathological changes in the colon tissue of CAC mice post-WMP intervention. Subsequently, differential genes and metabolites in the colorectal tissue of CAC mice following WMP intervention were identified through transcriptomics and non-targeted metabolomics. Finally, the influence of WMP on the peroxisome proliferator activated receptor (PPAR) pathway, Wnt pathway, and CC motif chemokine ligand 3 (CCL3)/ CC motif chemokine receptor 1 (CCR1) axis in CAC mice was verified through western blot, immunofluorescence, and ELISA based on the results of transcriptomics and non-targeted metabolomics. RESULTS: WMP intervention enhanced survival, alleviated body weight loss, shortened colon length, tumor occurrence, and pathological changes in the colorectal tissue of CAC mice, such as glandular damage, tumourigenesis, and inflammatory cell infiltration. Transcriptomic and non-targeted metabolomic results revealed that WMP intervention up-regulated the expression of key regulatory mechanisms of fatty acid oxidation PPAR pathway-related genes (Pparg, Ppara, Cpt1a, and Acadm) and metabolites (L-carnitine and L-palmitoylcarnitine). Additionally, it down-regulated Wnt pathway-related genes (Wnt3, Axin2, Tcf7, Mmp7, Lgr5, Wnt5a, Fzd6, Wnt7b, Lef1, and Fzd10 etc.) and pro-inflammatory related genes (Il1b, Il6, Il17a, Ccl3, and Ccr1 etc.). Experimental validation demonstrated that WMP up-regulated PPAR pathway-related proteins [PPARγ, PPARα, carnitine palmitoyltransferase 1A (CPT1A), and acyl-CoA dehydrogenase medium chain (ACADM)] in the colorectal tissue of CAC mice. It also down-regulated Wnt pathway-related proteins [ß-catenin, T-cell factor (TCF), lymphoid enhancer-binding factor (LEF), and matrix metallopeptidase 7 (MMP7)], inhibited the nuclear translocation of the key transcription factor ß-catenin in the Wnt pathway, and suppressed epithelial-to-mesenchymal transition (EMT) activation induced by the Wnt pathway (up-regulated E-cadherin and down-regulated Vimentin). Furthermore, WMP intervention reduced pro-inflammatory factors [interleukin (IL)-6, IL-1ß, and IL-17A] and decreased CCL3/CCR1 axis factors, including CCL3 protein levels and diminished F4/80+CCR1+ positive expressed cells. CONCLUSION: WMP significantly inhibits CAC tumorigenesis by up-regulating PPARα-mediated fatty acid oxidation, inhibiting the Wnt signaling pathway-mediated EMT, and suppressing CCL3/CCR1-mediated inflammatory responses.

Network pharmacology and experimental validation to elucidate the pharmacological mechanisms of Bushen Huashi decoction against kidney stones.

Introduction: Kidney stone disease (KS) is a complicated disease with an increasing global incidence. It was shown that Bushen Huashi decoction (BSHS) is a classic Chinese medicine formula that has therapeutic benefits for patients with KS. However, its pharmacological profile and mechanism of action are yet to be elucidated. Methods: The present study used a network pharmacology approach to characterize the mechanism by which BSHS affects KS. Compounds were retrieved from corresponding databases, and active compounds were selected based on their oral bioavailability (≥30) and drug-likeness index (≥0.18). BSHS potential proteins were obtained from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, whereas KS potential genes were obtained from GeneCards and OMIM, TTD, and DisGeNET. Gene ontology and pathway enrichment analysis were used to determine potential pathways associated with genes. The ingredients of BSHS extract were identified by the ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass spectrometry (UHPLC-Q/Orbitrap MS). The network pharmacology analyses predicted the potential underlying action mechanisms of BSHS on KS, which were further validated experimentally in the rat model of calcium oxalate kidney stones. Results: Our study found that BSHS reduced renal crystal deposition and improved renal function in ethylene glycol(EG)+ammonium chloride(AC)-induced rats, and also reversed oxidative stress levels and inhibited renal tubular epithelial cell apoptosis in rats. BSHS upregulated protein and mRNA expression of E2, ESR1, ESR2, BCL2, NRF2, and HO-1 in EG+AC-induced rat kidney while downregulating BAX protein and mRNA expression, consistent with the network pharmacology results. Discussion: This study provides evidence that BSHS plays a critical role in anti-KS via regulation of E2/ESR1/2, NRF2/HO-1, and BCL2/BAX signaling pathways, indicating that BSHS is a candidate herbal drug for further investigation in treating KS.

Efficacy and safety of thread embedding acupuncture for facial expression muscles atrophy after peripheral facial paralysis: study protocol for a randomized controlled trial.

BACKGROUND: Facial expression muscles atrophy is one kind of sequelae after peripheral facial paralysis. It causes critical problems in facial appearance of patient as well as social and psychological problems. This study aims to evaluate the efficacy and safety of Thread-embedding acupuncture (TEA) for the management of facial expression muscles atrophy after peripheral facial paralysis. METHODS: This is a patient-assessor blinded, randomized, sham-controlled trial. A total of fifty-six eligible patients will be randomly divided into TEA (n=28) and sham TEA (STEA) (n=28) groups. Both groups will receive TEA or STEA treatment at the frontal muscle and the depressor anguli oris muscle, at one predefined points once a week for eight weeks. Additionally, both groups will receive traditional acupuncture treatment at ten acupoints (GB20, LI4, LR3, GB12, ST7, SI18, LI20, BL2, SJ23, ST4) twice a week for eight weeks as a concurrent treatment. B-mode ultrasonography will be used to assess the changes in facial expression muscle thickness ratio of the affected/healthy side at baseline and at 10 weeks after screening, as the primary outcome. House-Brackmann Grade and lip mobility score will be measured and analyzed at baseline and 4, 8, 10, and 12 weeks after screening, as secondary outcomes. DISCUSSION: The study will compare TEA with sham TEA to explore the feasibility for TEA in improving facial expression muscles atrophy after peripheral facial paralysis. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR1900027170. Registered on 3 November 2019, http://www.chictr.org.cn/edit.aspx?pid=45173&htm=4.

[Acupoint application of herbal paste relieves symptoms and naso-mucosal inflammatory response by down-regulating serum TGF-ß1 in allergic rhinitis rats].

OBJECTIVE: To observe the effect of acupoint application of herbal paste on symptoms of allergic rhinitis (AR), serum immunoglobulin E (IgE) and transforming growth factor beta 1 (TGF-ß1) level, and number of nasal eosinophils (EOS) in rats with AR, so as to explore its underlying mechanisms. METHODS: Forty male Wistar rats were randomly divided into normal control, model, medication and acupoint application groups (n＝10 rats per group). The AR model was established by intraperitoneal (i.p.) injection of mixture solution of ovalbumin, aluminum hydroxide and normal saline (once every other day, for 7 times), and nasal drip plus spray inhalation of ovalbumin (on the following day of i．p., once daily for 9 days). For acupoint application, the prepared herbal paste (containing White Mustard Seed, Rhizoma Corydalis, unprocessed Radix Kansui, Herba Asari and ginger juice) was applied to bilateral "Feishu" (BL13), "Pishu" (BL20) and "Shenshu" (BL23) for 2 h, once every other day for 7 times. The rats in the medication group were given Fluticasone Propionate nasal spray daily for 14 days. Scores of nasal itching, sneezing and nasal discharge on the day after modeling and the ending of the intervention were used to evaluate behavioral changes. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of serum IgE and TGF-ß1, and the infiltration state of EOS in the nasal mucosa tissue was observed under light microscope after HE staining. RESULTS: After modeling and compared with the normal control group, the behavioral scores and the levels of serum IgE and TGF-ß1 were significantly higher (P<0.05), and the infiltration state of EOS got worse. Compared with the model group, the increased behavioral score and serum IgE and TGF-ß1 levels were evidently suppressed (P<0.05) and EOS infiltration severity in the nasal mucosa was obviously milder in both medication and acupoint application groups. No significant differences were found between the medication and acupoint application groups in behavioral score and serum IgE and TGF-ß1 levels (P>0.05). CONCLUSION: Acupoint application can improve the symptoms of AR rats, which may be associated with its effect in down-regulating the levels of serum IgE and TGF-ß1.

In Vivo Study on Site of Action of Sinapine Thiocyanate following Acupoint Herbal Patching.

OBJECTIVE: To investigate the site of action of sinapine thiocyanate (ST), following acupoint herbal patching (AHP). METHODS: Twenty Wistar rats were randomized into five groups (groups A, B, C, D, and E), and all groups received the same AHP in vivo. Skin samples were excised at 2 h, 4 h, 6 h, 10 h, and 26 h after AHP administration from group A to group E separately and the concentrations of ST in the skin were determined using a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method. A pharmacokinetic profile of ST following AHP was performed at the same time in a group of five Wistar rats to detect plasma levels at the same time intervals. RESULTS: The mean ± SD ST concentrations (ng/ml) at 2 h (group A), 4 h (group B), 6 h (group C), 10 h (group D), and 26 h (group E) after AHP administration were 250.01 ± 61.99, 61.01 ± 30.41, 40.12 ± 26.94, 78.66 ± 59.43, and 19.55 ± 18.95, respectively. No ST was detected in rats' plasma samples at the same time points. CONCLUSIONS: The site of action of ST following AHP is in the skin.

Effect and safety of stimulating acupoints in children with cough variant asthma: A Meta-analysis.

OBJECTIVE: To evaluate the efficacy and safety of stimulating acupoints in subjects with childhood cough variant asthma (CVA). METHODS: A Meta-analysis of randomized controlled trials (RCTs) about the comparison between acupoint sticking therapy and non-acupoint sticking therapy for childhood CVA was conducted. The trials' quality and risk bias were assessed using the Cochrane Handbook tool. Odds ratio (OR) or risk ratio (RR) with 95% confidence intervals (CIs), mean differences (MDs) or standardized mean differences (SMDs) of a random-effects model were calculated. Heterogeneity was assessed by P value and I 2 statistics. RESULTS: Thirteen studies were included in our review, indicating that the total effective rate of stimulating-acupoint group is better than that of control group [RR 1.19, 95% CI (1.13-1.26), P < 0.000 01]. The recurrence rate in two years [RR 0.31, 95% CI (0.19-0.51), P < 0.000 01] and cough duration [MD = -2.42, 95% CI (-3.75, -1.09), P = 0.0004] of childhood CVA in stimulating-acupoint group were significantly lower than those in control group. Besides, stimulating acupoints can reduce the level of IgE [SMD = -0.75, 95% CI (-1.21,-0.30), P = 0.001] and EOS [SMD = -0.36, 95% CI (-0.92, 0.21), P = 0.22]. CONCLUSION: Our findings suggest that stimulating acupoints had positive effects on childhood CVA and was relatively safe treatment. However, more RCTs with more useful indicators are warrant to confirm the current findings.