Corrigendum: A novel Chinese herbal and corresponding chemical formula for cancer treatment by targeting tumor maintenance, progression, and metastasis.

[This corrects the article DOI: 10.3389/fphar.2022.907826.].

Effect of electrode configuration in electroacupuncture on ischemic stroke treatment in rats.

Background and aim: This study investigated the effect of the electrode configuration on EA treating ischemic stroke. Experimental procedure: An ischemic stroke rat model was established. In the EA-P group, the anodes of EA were placed on the BL7 and BL8 acupoints of the lesioned, and the cathodes were placed on the BL7 and BL8 acupoints of the nonlesioned hemispheres; by contrast, in the EA-N group. Results: The difference in neurological deficit scores between the first and fourth days and the difference in Rotarod test time between the fourth and first days after reperfusion were greater in the EA-P and EA-N groups than in the sham group (all p < 0.001). In the lesioned hemisphere, neuronal nuclei (NeuN), γ-aminobutyric acid-A (GABA)-A, postsynaptic density 95 (PSD95), and astrocyte glutamate transporter 1 (GLT-1) expression and microtubule-associated protein 2 (MAP2)/glyceraldehyde 3-phosphate dehydrogenase (GADPH) ratios were greater and the glial fibrillary acid protein (GFAP)/GADPH ratios were smaller in the EA-P than in the sham group (all p < 0.05), but these ratios in the EA-N group were similar to those in the sham group (all p > 0.05); serum adrenaline and serotonin levels in the sham group were lower than those in the normal and EA-P groups (both p < 0.05), and cerebrospinal fluid (CSF) glutamate levels were higher in the EA-P group than in the sham group (p < 0.05). Conclusion: EA improved neurological function through multiple pathways. However, placing the anode on the lesioned hemisphere can provide more neuroprotection.

Acupuncture modulates development of myopia by reducing NLRP3 inflammasome activation via the dopamine-D1R signaling pathway.

BACKGROUND: Dopamine has been suggested to be a stop signal for eye growth and affects the development of myopia. Acupuncture is known to increase dopamine secretion and is widely used to treat myopia clinically. OBJECTIVE: The aim of this study was to determine if acupuncture inhibits myopia progression in form deprived Syrian hamsters by inducing rises in dopamine content that in turn suppress inflammasome activation. METHODS: Acupuncture was applied at LI4 and Taiyang every other day for 21 days. The levels of molecules associated with the dopamine signaling pathway, inflammatory signaling pathway and inflammasome activation were determined. A dopamine agonist (apomorphine) was used to evaluate if activation of the dopaminergic signaling pathway suppresses myopia progression by inhibiting inflammasome activation in primary retinal pigment epithelial (RPE) cells. A dopamine receptor 1 (D1R) inhibitor (SCH39166) was also administered to the hamsters. RESULTS: Acupuncture inhibited myopia development by increasing dopamine levels and activating the D1R signaling pathway. Furthermore, we also demonstrated that nucleotide-binding oligomerization domain (NOD)-, leucine-rich repeat (LRR)- and pyrin domain-containing protein 3 (NLR) family pyrin domain-containing 3 (NLRP3) inflammasome activation was inhibited by activation of the D1R signaling pathway. CONCLUSION: Our findings suggest that acupuncture inhibits myopia development by suppressing inflammation, which is initiated by activation of the dopamine-D1R signaling pathway.

Changes in Acupuncture-Induced Specific Acupoint Neurotransmitters are Possibly Related to Their Physiological Functions in Rats.

This study investigated changes in neurotransmitters induced by the application of electroacupuncture (EA) at Zusanli (ST36) and Neiguan (PC6). A total of 30 rats were divided into five groups: sham, ST (EA at bilateral ST36 and ST37), ScT (ST plus previous neurectomy of the bilateral sciatic nerves), ScS (sham plus previous neurectomy of the bilateral sciatic nerve), and PC (EA at bilateral PC6 and PC7). The P2X2 receptor expression was stronger in the sham group than in the ST and PC groups (both p < 0.05) but similar between the sham and ScT groups (p > 0.05). Dopamine levels in the extracellular fluid surrounding the acupoints were higher in the PC group than in the sham and ST groups during the postacupuncture period (both p < 0.05). Glutamate levels in the extracellular fluid surrounding the acupoints were higher in the ST group than in the sham group during the acupuncture period (p < 0.05) and higher in the ST group than in the sham and PC groups during the postacupuncture period (both p < 0.05). Serum adrenaline and noradrenaline levels were higher in the PC group than in the sham, ST, and ScT groups (all p < 0.05). Glutamate levels in the CSF were higher in the ST group than in the sham, ScS, and PC groups (all p < 0.05). GABA levels in the CSF were higher in the ST group than in the sham, ScT, and PC groups (all p < 0.05). EA at ST36 and ST37 and PC6 and PC7 exerted an analgesic effect, EA at PC6 and PC7 can enhance heart function, and EA at ST36 and ST37 modulates the cerebral cortex. However, the study needs an evaluation of direct pain behavior, heart function, and brain function in the future.

A Novel Chinese Herbal and Corresponding Chemical Formula for Cancer Treatment by Targeting Tumor Maintenance, Progression, and Metastasis.

We characterized a so-called "heirloom recipe" Chinese herbal formula (temporarily named Formula X) that contains five Chinese medical botanical drugs, Huang-Lian (Coptis chinensis Franch. [Ranunculaceae]), Huang-Qin (Scutellaria baicalensis Georgi [Lamiaceae]), Bai-Wei (Vincetoxicum atratum (Bunge) C. Morren and Decne. [Apocynaceae]), E-Zhu (Curcuma aromatica Salisb. [Zingiberaceae]) and Bai-Zhu (Atractylodes macrocephala Koidz. [Asteraceae]). Formula X inhibited the growth of various cancer cells and decreased the expression levels of a panel of proteins, including CD133, Myc, PD-L1, and Slug, in cancer cells. We further found that the inhibition of growth and protein expression were exerted by Huang-Lian, Huang-Qin, and Bai-Wei (formula HHB), which exhibited the same biological effects as those of Formula X. Furthermore, we selected three active chemicals, berberine, baicalin, and saponin from Huang-Lian, Huang-Qin, and Bai-Wei, respectively, to produce a chemical formulation (formula BBS), which exhibited similar effects on cell growth and protein expression as those induced by formula HHB. Both the formulae HHB and BBS suppressed tumor growth in an animal study. Moreover, they decreased the protein levels of Myc and PD-L1 in tumor cells in vivo. In summary, we established a novel Chinese herbal formula and a chemical formula that targeted three important processes, tumor maintenance (tumor stem cells), progression, and metastasis, and that influenced the response of tumors to host immunosuppression, for the potentially effective treatment of cancer patients.

Effect of the traditional Chinese herb Helminthostachys zeylanica on postsurgical recovery in patients with ankle fracture: A double-blinded randomized controlled clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Helminthostachys zeylanica (HZ), which is also called "Dao-Di-U-Gon" in Taiwan, has anti-inflammatory and antiedema effects and is commonly used to treat edema in patients with fractures. The ugonin K component of HZ can induce osteogenesis and promote bone mineralization, its therapeutic effect, however, its therapeutic effect remains unclear. Therefore, the purpose of the present study was to investigate the effect of HZ on functional recovery in patients with ankle fractures requiring surgical treatment. METHODS: A double-blinded, randomized, controlled study was conducted. A total of 45 patients with ankle fractures requiring surgical treatment were assigned to either the control group (n = 23 patients), which received the oral administration of HZ placebo 1.0 g t.i.d. for 42 days continuously, or to the treatment group (22 patients), which received HZ for 42 days. RESULTS: The serum amino-terminal propeptide of type 1 procollagen (PINP) levels were similar in the first assessment (V1) between the control (45.90 ± 16.31 ng/mL) and treatment groups (52.61 ± 21.02 ng/mL; p = 0.240); the differences in PINP level between the third assessment (V3) and V1 were greater in the treatment group (35.84 ± 24.56 ng/mL) than in the control group (16.34 ± 11.97 ng/mL; p = 0.002). Radiographic healing time (RHT) was 9.09 ± 1.15 weeks in the treatment group, which was shorter than the 9.91 ± 0.79 weeks (p = 0.012) in the control group. CONCLUSION: Oral administration of HZ for 42 days can increase serum PINP level and reduce the RHT. Therefore, HZ can be used to treat patients with ankle fractures requiring surgical treatment. However, a larger sample size is needed in future studies.

Effects of Yi-Gan-san on the psychiatric behavior of children and adolescents with Tourette's Syndrome: A randomized, double-blind, controlled preliminary study.

ETHNOPHARMACOLOGICAL RELEVANCE: Gilles de la Tourette's Syndrome (TS) is a childhood-onset disease with clinical features of motor and phonic tics. Yi-Gan-san (YGS) is a traditional Chinese medicine formula that can reduce aggressiveness and agitation and inhibit dopamine function. This study investigated the effects of YGS on the psychiatric behavior of children and adolescents with TS. METHODS: A double-blind, randomized, controlled preliminary study was conducted. A total of 38 patients with TS were assigned to the control group (CG, 19 patients) who received the oral administration of YGS placebo (90% starch and 10% YGS; 2.5 g thrice daily) or to a treatment group (TG, 19 patients) who received YGS for 4 weeks. The primary outcome measure was the change in Yale Global Tic Severity Scale (YGTSS) overall and subscale scores. RESULTS: The intensity score for phonic tics before oral administration of YGS, and after 2 weeks, 3 weeks and 4 weeks was not significantly different between CG and TG groups (2.94 ± 1.14 vs 2.79 ± 1.08, p = .686; 2.29 ± 1.21 vs 1.95 ± 1.08, p = .370; 2.41 ± 1.18 vs 2.05 ± 1.51, p = .435; and 2.29 ± 1.26 vs 1.84 ± 1.42, p = .323, respectively), while the intensity score for phonic tics after 1-week oral administration of YGS in the TG was 1.89 ± 1.10 lower than 3.06 ± 1.39 in the CG (p = .008). CONCLUSION: Oral administration of YGS for 1 week only reduced the intensity of phonic tics compared with oral administration of YGS placebo, suggesting that YGS can reduce their intensity for a short period, and the compliance of oral administration of YGS for 4 weeks can be accepted in children and adolescents with Tourette's Syndrome. However, because this study was preliminary, the selection of an appropriate placebo and dosage and long-term observations are crucial areas for future studies.

Metabolism modulation in rat tissues in response to point specificity of electroacupuncture.

Zusanli (ST36) and Neiguan (PC6) are acupoints along two meridians. To demonstrate point specificity, we investigated the effects of ST36 and PC6 in electroacupuncture (EA)-treated rats. The rats were subjected to sham acupuncture at ST36 without electric stimulation, EA at ST36, or EA at PC6. Heart and stomach tissues were collected for metabolite profiling. Each type of stimulation resulted in a different metabolite composition in the rat heart and stomach tissues. In the heart tissues, EA at ST36 affected a wider range of metabolite pathways than did EA at PC6, whereas similar numbers of metabolites in the stomach tissues were affected by EA at ST36 and PC6. The pathways affected by EA at ST36 differed from those affected by EA at PC6, and a group of common metabolites were reversely regulated by these two acupoints. This study demonstrated point specificity effectively modulated metabolism in rat heart and stomach tissues. The results indicate that heart stimulation may be connected to the stomach through the pericardium meridian (as described in traditional Chinese medicine), explaining why acupuncture applied to the stomach meridian can be an alternative treatment for gastric and heart diseases.

Evaluations and Mechanistic Interrogation of Natural Products Isolated From Paeonia suffruticosa for the Treatment of Inflammatory Bowel Disease.

Mu Dan Pi (MDP), a traditional Chinese medicine derived from the root bark of Paeonia suffruticosa Andrews, is used to treat autoimmune diseases due to its anti-inflammatory properties. However, the impact of MDP on inflammatory bowel disease (IBD) and its principal active compounds that contribute to the anti-inflammatory properties are uncertain. Thus, this study systemically evaluated the anti-inflammatory effects of fractionated MDP, which has therapeutic potential for IBD. MDP fractions were prepared by multistep fractionation, among which the ethyl acetate-fraction MDP5 exhibited the highest potency, with anti-inflammatory activity screened by the Toll-like receptor (TLR)-2 agonist, Pam3CSK4, in a cell-based model. MDP5 (at 50 µg/ml, p < 0.001) significantly inhibited nuclear factor kappa-B (NF-κB) reporters triggered by Pam3CSK4, without significant cell toxicity. Moreover, MDP5 (at 10 µg/ml) alleviated proinflammatory signaling triggered by Pam3CSK4 in a dose-dependent manner and reduced downstream IL-6 and TNF-α production (p < 0.001) in primary macrophages. MDP5 also mitigated weight loss, clinical inflammation, colonic infiltration of immune cells and cytokine production in a murine colitis model. Index compounds including paeoniflorin derivatives (ranging from 0.1 to 3.4%), gallic acid (1.8%), and 1,2,3,4,6-penta-O-galloyl-ß-D-glucose (1.1%) in MDP5 fractions were identified by LC-MS/MS and could be used as anti-inflammatory markers for MDP preparation. Collectively, these data suggest that MDP5 is a promising treatment for IBD patients.

Auricular Acupuncture to Lower Blood Pressure Involves the Adrenal Gland in Spontaneously Hypertensive Rats.

Auricular acupuncture is used to treat cardiac-related diseases such as hypertension. Therefore, the purpose of the present study was to investigate the effects of auricular acupuncture on blood pressure (BP) in spontaneously hypertensive rats (SHRs). The treatment group (TG) received auricular electroacupuncture (EA) at the auricle heart (CO15) and auricle shenmen (TEF3) points. Heart rate (HR) and BP, GABA-A expression, catecholamine, and neurotransmitter levels were measured. The HR was reduced after 7 auricular EA treatments compared with controls (all p < 0.05). Systolic BP and diastolic BP also decreased immediately and throughout the treatments compared with controls (all p < 0.05). The reduction of BP and HR was reversed by bicuculline injection 30 min before auricular EA treatment (all p < 0.05). GABA levels in the adrenal gland were higher with auricular EA treatment compared with the control group at 4 h (p < 0.05). Levels of serum noradrenaline and adrenaline were reduced at 15 min after final auricular EA treatment compared with the normal control group (both p < 0.05). The lowering of BP and HR by auricular EA is possibly mediated via vagal afferents from the concha to the nucleus of the solitary tract. After signal integration in the medulla oblongata, it may be transmitted through sympathetic efferent or vagal efferent or through multiple signaling pathways simultaneously to the atrionector of heart and the adrenal medulla. Further study is warranted.

Docosahexaenoic acid inhibits the proliferation of Kras/TP53 double mutant pancreatic ductal adenocarcinoma cells through modulation of glutathione level and suppression of nucleotide synthesis.

The treatment of cancer cells obtained by blocking cellular metabolism has received a lot of attention recently. Previous studies have demonstrated that Kras mutation-mediated abnormal glucose metabolism would lead to an aberrant cell proliferation in human pancreatic ductal adenocarcinoma (PDAC) cells. Previous literature has suggested that consumption of fish oil is associated with lower risk of pancreatic cancer. In this study, we investigated the anti-cancer effects of docosahexaenoic acid (DHA) in human PDAC cells in vitro and in vivo. Omega-3 polyunsaturated fatty acids (PUFAs) such as DHA and eicosapentaenoic acid (EPA) significantly inhibited the proliferation of human PDAC cells. The actions of DHA were evaluated through an induction of cell cycle arrest at G1 phase and noticed a decreased expression of cyclin A, cyclin E and cyclin B proteins in HPAF-II cells. Moreover, it was found that co-treatment of DHA and gemcitabine (GEM) effectively induced oxidative stress and cell death in HPAF-II cells. Interestingly, DHA leads to an increased oxidative glutathione /reduced glutathione (GSSG/GSH) ratio and induced cell apoptosis in HPAF-II cells. The findings in the study showed that supplementation of GSH or N-Acetyl Cysteine (NAC) could reverse DHA-mediated cell death in HPAF-II cells. Additionally, DHA significantly increased cellular level of cysteine, cellular NADP/NADPH ratio and the expression of cystathionase (CTH) and SLCA11/xCT antiporter proteins in HPAF-II cells. The action of DHA was, in part, associated with the inactivation of STAT3 cascade in HPAF-II cells. Treatment with xCT inhibitors, such as erastin or sulfasalazine (SSZ), inhibited the cell survival ability in DHA-treated HPAF-II cells. DHA also inhibited nucleotide synthesis in HPAF-II cells. It was demonstrated in a mouse-xenograft model that consumption of fish oil significantly inhibited the growth of pancreatic adenocarcinoma and decreased cellular nucleotide level in tumor tissues. Furthermore, fish oil consumption induced an increment of GSSG/GSH ratio, an upregulation of xCT and CTH proteins in tumor tissues. In conclusion, DHA significantly inhibited survival of PDAC cells both in vitro and in vivo through its recently identified novel mode of action, including an increment in the ratio of GSSG/GSH and NADP/NADPH respectively, and promoting reduction in the levels of nucleotide synthesis.

A systematic identification of anti-inflammatory active components derived from Mu Dan Pi and their applications in inflammatory bowel disease.

Mu Dan Pi (MDP), also known as Moutan Cortex Radicis, is a traditional Chinese medicine used to treat autoimmune diseases. However, the impact of MDP and its principal active compounds on inflammatory bowel disease (IBD) is uncertain. This study therefore systemically assessed the anti-inflammatory effects of MDP and its known active compounds in IBD. The anti-inflammatory activities of water extract and individual compounds were screened by NF-κB and interferon regulatory factor (IRF) reporter assays in THP-1 cells induced with either Toll-like receptor or retinoic acid inducible gene I/melanoma differentiation-associated gene 5 activators and further verified in bone marrow-derived macrophages. MDP water extract significantly inhibited the activation of NF-κB and IRF reporters, downstream signaling pathways and the production of IL-6 and TNF-α, in a dose-dependent manner. Among 5 known active components identified from MDP (1,2,3,4,6-penta-O-galloyl-ß-d-glucose [PGG], gallic acid, methyl gallate, paeoniflorin, and paeonol), PGG was the most efficient at inhibiting both reporters (with an IC50 of 5-10 µM) and downregulating IL-6 and TNF-α. Both MDP powder for clinical use and MDP water extract, but not PGG, reduced colitis and pathological changes in mice. MDP and its water extract show promise as a novel therapy for IBD patients.

Electroacupuncture at Zusanli and at Neiguan characterized point specificity in the brain by metabolomic analysis.

Different point stimulations can induce brain activity in specific regions, and however whether these stimulations affect unique neurotransmitter transmission remains unknown. Therefore, we aimed to investigate the effect of point specificity to the brain by resolving the metabolite profiles. Eighteen Sprague-Dawley rats were randomly divided into three groups: (1) the sham group: sham acupuncture at Zusanli (ST36) without electric stimulation; (2) the Zusanli (ST36) group: electroacupuncture (EA) at ST36; and (3) the Neiguan (PC6) group: EA at PC6. Then, the metabolites from rat brain samples were measured by LC-ESI-MS. The results of a partial least squares discriminant analysis revealed the differences among the sham, ST36, and PC6 groups regarding the relative content of metabolites in the cerebral cortex, hippocampus, and hypothalamus. EA at PC6 resulted in downregulation of adenosine, adrenaline, γ-aminobutyric acid, glycine, and glutamate majorly in hippocampus, and then in cerebral cortex. Otherwise, EA at ST6 resulted in upregulation of adrenaline and arginine in hippocampus, and all stimulations showed barely change of identified neurotransmitters in hypothalamus. These differential metabolite and neurotransmitter profiles prove that brain areas can be modulated by point specificity and may provide a maneuver to understand more details of meridian.

Electroacupuncture Relieves CCI-Induced Neuropathic Pain Involving Excitatory and Inhibitory Neurotransmitters.

Neuropathic pain caused by peripheral tissue injuries to the higher brain regions still has no satisfactory therapy. Disruption of the balance of excitatory and inhibitory neurotransmitters is one of the underlying mechanisms that results in chronic neuropathic pain. Targeting neurotransmitters and related receptors may constitute a novel approach for treating neuropathic pain. We investigated the effects of electroacupuncture (EA) on chronic constriction injury- (CCI-) induced neuropathic pain. The mechanical allodynia and thermal hyperalgesia pain behaviors were relieved by 15 Hz EA but not by 2 and 50 Hz. These phenomena were associated with increasing γ-amino-butyric acid (GABA) receptors in the hippocampus and periaqueductal gray (PAG) but not N-methyl-D-aspartate receptors. Furthermore, excitatory neurotransmitter glutamate was decreased in the hippocampus and inhibitory neurotransmitter GABA was increased in the PAG under treatment with EA. These data provide novel evidence that EA modulates neurotransmitters and related receptors to reduce neuropathic pain in the higher brain regions. This suggests that EA may be a useful therapy option for treating neuropathic pain.

Aqueous extracts of Paeonia suffruticosa modulates mitochondrial proteostasis by reactive oxygen species-induced endoplasmic reticulum stress in pancreatic cancer cells.

BACKGROUND: Pancreatic cancer (PC) remains the leading cause of cancer mortality, with limited therapeutic targets, and alterations in endoplasmic reticulum (ER)-related proteostasis may be a potential target for therapy. The root bark of Paeonia suffruticosa has been shown to inhibit cancer growth and metastasis, although its impact on PC is unknown. PURPOSE: To ascertain the anti-cancer effects of P. suffruticosa on oncogenic functions of PC and determine the detailed molecular mechanisms. STUDY DESIGN: Efficacy assessment of extracts, in vitro using PC cells as a model system and in vivo in mouse xenograft tumors. METHODS: P. suffruticosa aqueous extracts (PS) were prepared and assessed using liquid chromatography-tandem mass spectrometry. Cell viability, proteins, and cell components were measured using MTT assay, western blotting, and immunofluorescence. Cell apoptosis, cell cycle, and migration were assessed using colorimetric assays, fluorescence activated cell sorting, and transwell migration. Reactive oxygen species (ROS) were evaluated with a commercial 2'-7'-dichlorofluorescin diacetate kit. For the xenograft model, AsPC1 cells were inoculated subcutaneously into immunocompromised mice and PS (oral) was administered over 3 weeks with or without gemcitabine (GEM, intraperitoneal), a first-line advanced/metastatic PC therapy. RESULTS: PS stimulated ER stress and affected mitochondrial membrane potential to increase autophagosome numbers and block their degradation, followed by autophagy induction and finally cell apoptosis. Additionally, PS-mediated proteostasis impairment resulted in altered dynamics of the actin cytoskeleton, cell motility impairment, and cell cycle progression inhibition. Conversely, a ROS scavenger partially reversed PS-mediated degradation of peptidyl-prolyl cis-trans isomerase B (PPIB), an ER protein important for protein folding, suggesting that ROS generation by PS may be the upstream of PS-triggering of mitophagy and final cell apoptosis. Nevertheless, oral administration of PS, alone or in combination with GEM, delayed tumor growth in a xenograft model without affecting body weight. CONCLUSION: These findings indicate that PS may constitute a potential new alternative or complementary medicine for PC.