Anti-hypertensive and endothelia protective effects of Fufang Qima capsule on primary hypertension via adiponectin/adenosine monophosphate activated protein kinase pathway.

OBJECTIVE: To investigate the potential mechanism of the vascular remodeling effect and provide additional information about anti-hypertension activity of Fufang Qima capsule. METHODS: Spontaneous hypertensive rats (SHRs) were used to study the underlying mechanism of the anti-hypertension activity of QM. In this study, SHRs were randomly divided into 5 groups: model group, Telmisartan group (7.2 mg/kg, p.o.), and three QM groups (0.9298, 1.8596, and 3.7192 g/kg, p.o.). Wistar Kyoto rats (WKY) were used as normal control group. Blood pressure (BP), aorta, perivascular adipose tissue (PVAT) histology were investigated to evaluate the effect of QM. Nitric oxide (NO) and endothelial nitric oxide synthase (eNOS) phosphorylation were measured. Adiponectin (APN) secretion, as well as APN signal pathway proteins including APN, adiponectin receptors (R1 and R2) and adenosine 5'-monophosphate-activated protein kinase (AMPK) were all analyzed. RESULTS: QM significantly reduced BP and ameliorated the vascular pathological change, i.e. intima media thicken and collagen fiber hyperplasia. Meanwhile, QM increased concentration of NO and the phosphorylation of eNOS in the aorta. The anti-hypertensive and endothelia-protective effect of QM could be attributed to activating APN/ AMPK pathway by up-regulating the expression of APN in PVAT and APN Receptor 2, AMPKα and phosphorylated AMPKα in the aorta. CONCLUSION: The QM alleviation effect mechanism for primary hypertension was via modulating the APN/AMPK signal pathway.

Neuroprotective Effect of Fructus broussonetiae on APP/PS1 Mice via Upregulation of AKT/ß-Catenin Signaling.

OBJECTIVE: To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae (FB) in both mouse and cell models of Alzheimer's disease (AD). METHODS: APP/PS1 mice treated with FB for 2 months and vehicle-treated controls were run through the Morris water maze and object recognition test to evaluate learning and memory capacity. RNA-Seq, Western blotting, and immunofluorescence staining were also conducted to evaluate the effects of FB treatment on various signaling pathways altered in APP/PS1 mice. To further explore the mechanisms underlying FB's protective effect, PC-12 cells were treated with Aß25-35 in order to establish an in vitro model of AD. RESULTS: FB-treated mice showed improved learning and memory capacity on both the Morris water maze and object recognition tests. RNA-seq of hippocampal tissue from APP/PS1 mice showed that FB had effects on multiple signaling pathways, specifically decreasing cell apoptotic signaling and increasing AKT and ß-catenin signaling. Similarly, FB up-regulated both AKT and ß-catenin signaling in PC-12 cells pre-treated with Aß25-35, in which AKT positively regulated ß-catenin signaling. Further study showed that AKT promoted ß-catenin signaling via enhancing ß-catenin (Ser552) phosphorylation. Moreover, AKT and ß-catenin signaling inhibition both resulted in the attenuated survival of FB-treated cells, indicating the AKT/ß-catenin signaling is a crucial mediator in FB promoted cell survival. CONCLUSIONS: FB exerted neuroprotective effects on hippocampal cells of APP/PS1 mice, as well as improved cell viability in an in vitro model of AD. The protective actions of FB occurred via the upregulation of AKT/ß-catenin signaling.

Skin-patch of Xin Huang Pian on relieving joint symptoms in patients with acute gouty arthritis: A Randomized, Double-Blind, Active-Controlled Trial.

AIMS: To evaluate the effectiveness and safety of Xin Huang Pian skin patches for patients with acute gouty arthritis. BACKGROUND: In China, patients with acute gouty arthritis benefit from skin patcheses with herbal medicines. But the clinical effects of skin patches with Xin Huang Pian are rarely reported. DESIGN: A Randomized, Double-Blind, Active-Controlled Trial. METHODS: The trial was performed from January 2015-December 2018 at the First Affiliated Hospital of Sun Yat-sen University in China. It was conducted with one intervention group (skin patches of Xin Huang Pian, N = 30) and one active control group (skin patches of Diclofenac Diethylamine Emulgel, N = 31). Participants and study investigators were both blinded to the treatment assignments. The primary outcomes were the improvement of joints' symptoms. The secondary outcomes were changes in white blood cells, erythrocyte sedimentation rate and C-reactive protein. RESULTS: Skin patches of Xin Huang Pian showed quick effect on decreasing joint pain at 3rd day of treatment. Wherever only at 7th day, Diclofenac Diethylamine Emulgel markedly lowered joint pain. Xin Huang Pian also showed superior effect than Diclofenac Diethylamine Emulgel on improving joint swelling and range of motion and decreasing the levels of C-reactive protein and erythrocyte sedimentation rate. No adverse reactions were observed in skin patches of Xin Huang Pian treatment. CONCLUSION: Skin patches of Xin Huang Pian appeared to be safe and efficacious for relieving joint symptoms in patients with acute gouty arthritis. The mechanism might be associated with the decreased levels of C-reactive protein and erythrocyte sedimentation rate. IMPACT: Skin-patcheses with Xin Huang Pian are more effective than Diclofenac Diethylamine Emulgel on improving joint pain, swelling and range of motion. Xin Huang Pian treatment showed superior effects compared with Diclofenac Diethylamine Emulgel on decreasing levels of C-reactive protein and erythrocyte sedimentation rate. Patients with acute gouty arthritis may benefit from skin patches of Xin Huang Pian for effective relief from joint pain and swelling. Chinese Clinical Trial Registration: ChiCTR-TRC-1300 4122.

[Treatment of scapulohumeral periarthritis by Fuyang-pot warming combined with electroacupuncture stimulation].

OBJECTIVE: To observe the clinical therapeutic effect of Fuyang-pot warming combined with electroacupuncture (EA) in the treatment of scapulohumeral periarthritis (SPA). METHODS: A total of 90 cases of SPA patients were randomized into EA, Fuyang-pot warming and EA plus Fuyang-pot warming (combination) groups (n=30 per group). Fuyang-pot warming including pressing, mild moxibustion, scraping-pushing, cupping, tapping, etc. was applied to Fengchi (GB20), Dazhui (GV14), Jianjing (GB21), Jianyu (LI15), Zhongfu (LU1), Ashi-point, etc., and EA (2 Hz /100 Hz,1-1.5 mA) was appled to GB20, GV14, GB21, LI15, Binao (LI14), Tiaokou (ST38), Chengshan (BL57), Ashi-point, etc. The treatment was performed for 30 min every time, once every other day for 2 weeks. The visual analogue scale (VAS, 0-10 points) was used to assess the pain severity. The Constant-Murley shoulder assessment scale (100 points in total, including 15 points in pain severity and 20 points in daily living activities, 40 points in joint motion range, and 25 points in myodynamia) was used to assess the functional state of the shoulder. The rating scale of the American Shoulder and Elbow Surgeons (ASES, 4 grades) was used to evaluate the ability of daily living activities. RESULTS: Following the treatment, intra-group comparison showed that the VAS score was significantly reduced in the three groups in comparison with their own pre-treatment (P<0.01). The total scores of Constant-Murley scale, and scores of activities of daily living and active motion range, myodynamia, and ASES shoulder-joint function were all considerably increased in the three groups in comparison with their own pre-treatment (all P<0.01). The therapeutic effect of EA plus Fuyang-pot warming was significantly superior to that of simple EA and simple Fuyang-pot warming in reducing VAS score and increasing total score of Constant-Murley scale and scores of activities of daily living, active motion range, myodynamia as well as ASES shoulder joint function (P<0.01). Of the 30, 29 and 30 cases in the combination, EA and Fuyang-pot warming groups, 9, 2 and 4 were basically cured, 14, 8 and 12 experienced marked improvement, 4, 12 and 9 were improved, and 3, 7 and 5 failed in the treatment, with the cured plus effective rates being 76.67%, 34.48% and 53.33%, respectively. The cure plus effective rate was apparently higher in the combined treatment group than in the simple EA and simple Fuyang-pot warming groups (P<0.01), but had no significant difference between the simple EA and simple Fuyang-pot warming groups (P>0.05). CONCLUSION: Fuyang-pot warming combined with EA is effective in relieving pain, and enhancing the daily life quality in scapulohumeral periarthritis patients.

Evolution of matrinic ethanol derivatives as anti-HCV agents from matrine skeleton.

Twenty-two novel 12N-substituted matrinic ethanol derivatives were synthesized and evaluated for their antiviral activities against HCV taking compound 1 as the lead. The SAR study indicated that the shortening of the 11-butyl chain to ethyl chain did not affect the activity significantly. Out of the target compounds, matrinic ethanol 6a demonstrated a potential anti-HCV effect with an EC50 value of 3.2µM and a SI value of 96.6. The free hydroxyl arm in 6a made it possible as a parent structure to prepare pro-drug for the potential application in HCV treatment. This study provided powerful information on further strategic optimization and development of this kind of compounds into a novel family of anti-HCV agents.

Structure-activity relationship of N-benzenesulfonyl matrinic acid derivatives as a novel class of coxsackievirus B3 inhibitors.

A novel series of N-benzenesulfonyl matrinic amine/amide and matrinic methyl ether analogues were designed, synthesized and evaluated for their in vitro anti-coxsackievirus B3 (CVB3) activities. The structure-activity relationship (SAR) studies revealed that introduction of a suitable amide substituent on position 4' could greatly enhance the antivirus potency. Compared to the lead compounds, the newly synthesized matrinic amide derivatives 21c-d and 21j exhibited stronger anti-CVB3 activities with lower micromolar IC50 from 2.5 µM to 2.7 µM, and better therapeutic properties with improved selectivity index (SI) from 63 to 67. The SAR results provided powerful information for further strategic optimization, and these top compounds were selected for the next evaluation as novel enterovirus inhibitors.

Effect of Tiantai No.1 on gene expression profiles in hippocampus of Alzheimer's disease rats by bioinformatic analysis.

OBJECTIVE: To study the effect of Tiantai No. 1 [symbol in text] on gene expression profile in hippocampus of Alzheimer's disease (AD) rat, molecular genetic target points of the effect of this drug were defined, its molecular genetic pharmacodynamic mechanism of anti-AD was further explored at molecular gene level, and a scientific basis was provided for its clinical availability and promotion. METHODS: Thirty male Sprague-Dawley rats were divided into three groups with 10 rats per group: sham-operation group, model group and Tiantai No. 1 group. Sterile surgical procedure was applied, the model group with bilateral hippocampal injection of Aß1-40 was established, and normal saline was used instead of Aß1-40 in the sham-operation group. One week after the models was made, rats were administered by gastric lavage once every day for three consecutive weeks. The rats of the sham-operation group and the model group were daily fed with purified water by lavage; the rats of the Tiantai No.1 group treated group were administered with Tiantai No.1 by lavage. Total RNAs of hippocampus tissues were extracted with Trizol, the changes of hippocampus gene expression profiles in the above three groups were analyzed by using Affymetrix rat whole genome expression profile microarray. RESULTS: Microarray analysis showed that, compared with the sham-operation group, the hippocampus of the model group had 50 up-regulated genes with significant difference (fold change >2), and 21 down-regulated genes with significant difference (fold change <0.5); compared with the hippocampus of the model group, the hippocampus of the Tiantai No. 1 group was found to have 5 up-regulated genes with significant difference (fold change >2) and 20 down-regulated genes with significant difference (fold change <0.5). The functions of differentially expressed genes of the groups were involved in nervous system's development, neuronic differentiation and function-regulation, cellular growth and differentiation and apoptosis, synaptic occurrence and plasticity, inflammation and immune response, ion channels/transporters, cellular signal transduction, cellular material/energy metabolism and so on. CONCLUSION: Tiantai No. 1 can regulate hippocampal function, and further regulate the brain function of animals in multiple gene target points by a number of ways.

[Influence of different acupuncture manipulations at Zusanli (ST 36) on skin microcirculation blood perfusion in healthy subjects].

OBJECTIVE: To observe the influence of different acupuncture manipulations at Zusanli (ST 36) on blood perfusion of skin microcirculation in healthy volunteer subjects. METHODS: A total of 10 healthy volunteers were recruited in the present study. Acupuncture manipulations of lifting-thrusting reinforcing and reducing, uniform reinforcing-reducing, and simple needle insertion were applied to right Zusanli (ST 36) for 2 min. Changes of blood perfusion in the right Zusanli (ST 36) area were detected 1 min, 5 min, 10 min, 15 min, 20 min, 25 min and 30 min after acupuncture stimulation, respectively by using a Pericam Perfusion Speckle Imager. The interval between two detections for different needling manipulations was 2 days at least. RESULTS: In comparison with pre-acupuncture stimulation, the blood perfusion levels of the skin microcirculation around the right Zusanli (ST 36) region were significantly increased from 1 min to 10 min following simple needle insertion, from 5 min to 30 min after uniform reinforcing-reducing manipulation, from 1 min to 30 min after reinforcing manipulation, and from 1 min to 25 min following reducing manipulation, respectively (P < 0.05, P < 0.01), and the effects of reinforcing manipulation were apparently superior to those of the reducing, uniform reinforcing-reducing manipulations and simple needle insertion in upregulating blood perfusion level (P < 0.05, P < 0.01). CONCLUSION: Lifting-thrusting reinforcing, reducing, uniform reinforcing-reducing manipulations of acupuncture needle, and simple needle insertion at Zusanli (ST 36) can obviously increase blood perfusion level in normal volunteer subjects, and the effect of reinforcing manipulation is apparently better.

[Amplification effect of acupoint from feeling of acupoint and transmitting of meridian].

Through arrangement and analysis of domestic and overseas literature about effect of acupoint, it is found that acupoint is an amplifier of acupuncture effect, which could present feature of weak-stimulation and strong-response for acupuncture stimulation. It is also thought that factors of the amplification effect are complicated, it is closely related to the local specificity of acupoint (including more nerves, more blood vessels, more mast cells, more Ca2+ and more energy metabolism) and circulation route of meridians, indicating the amplification effect works through local feeling (local specificity of acupoint) into the distal end (circulation and distribution route of meridian system.

[A review of recent researches on correlation between ATP and acupuncture efficacies].

It has been documented that adenosine triphosphate (ATP) is a multifunctional nucleoside triphosphate used in cells, including chemical energy transportation, extra- and intracellular signaling, cell structure maintaining, DNA and RNA synthesis, etc. In the present paper, the authors reviewed studies on the involvement of ATP in different efficacies of acupuncture intervention from the following four aspects. 1) ATP release in the stimulated acupoint area is one of the key factors for producing acupuncture analgesia; 2) Acupuncture induced suppression of ATP activity in the central nervous system results in pain relief; 3) ATP application on the human body surface may strengthen the sensation propagation along the meridian; 4) Favorable regulation of acupuncture intervention on the abnormal functional activities of some viscera often accompanies with an increase of ATP content and ATPase activity in the related internal organs. It has been proposed that ATP, Ca2+ and reactive oxygen species (ROS) are closely related each other in the life activities of the organism. Hence, a reasonable regulation on ATP levels in the related organs of the body may be a new approach for raising clinical therapeutic effects of acupuncture therapy.

[Study on the feasibility for judging the lifting-thrusting manipulation of acupuncture by using "quality control figure" measurement].

OBJECTIVE: To explore the feasibility of judgment of the lifting-thrusting acupuncture manipulation by using "quality control figure" measurement, so as to provide a possible reference for demonstration in acupuncture teaching and scientific research. METHODS: A total of 13 high-professional title acupuncturists and teachers and 100 college students participated in the present study. The so-called "quality control figures" of lifting-thrusting acupuncture skill were established by collecting operation data from 13 acupuncturists and teachers with an "Acupuncture Manipulation Parameter Detector System". A total of 100 college students who have learnt acupuncture and massage learnings were asked to perform the same acupuncture manipulation after training, followed by giving a mark with conventional manipulation method and "quality control figure" method (scores), respectively. In addition, the correlation between these two scoring methods was analyzed. RESULTS: The average score of conventional scoring of lifting-thrusting manipulation was (81.03 +/- 18.40) points and that of "quality control figure" scoring was (80.60 +/- 7.48) points in those college students after training. No significant difference was found between these two scoring methods (P > 0.05). But a positive correlation was found between them (r = 0.6943, P < 0.01), suggesting a positive feasibility for judging the lifting-thrusting acupuncture manipulation with "quality control figure" measurement. CONCLUSION: A positive correlation exists between the conventional scoring and "quality control figure" scoring for lifting-thrusting acupuncture manipulation, suggesting a feasibility in judging this acupuncture manipulation with "quality control figure" measurement.

Endoplasmic reticulum stress induced by tunicamycin and antagonistic effect of Tiantai No.1 (1) on mesenchymal stem cells.

OBJECTIVE: Changes of the internal and external cellular environments can induce calcium homeostasis disorder and unfolded protein aggregation in the endoplasmic reticulum (ER). This ER function disorder is called endoplasmic reticulum stress (ERS). Severe long-term ERS can trigger the ER apoptosis signaling pathway, resulting in cell apoptosis and organism injury. Recent researches revealed that ERS-induced cell death was involved in the neurocyte retrogradation in the progress of neuron degenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease and so on. Therefore, the protection effect of the traditional Chinese drug-Tiantai No. 1 (1) on the ERS injury of AD was investigated at the molecular gene level in this study with a view to explore the gene pharmacodynamic actions and mechanisms of this drug. METHODS: Primarily cultured marrow mesenchymal stem cells (MSCs) of rats were treated by tunicamycin (TM) in order to induce ERS. RT-PCR, fluorescence immunocytochemistry and Western blot techniques were used to determine the mRNA and protein expression levels of the protective stress protein-ER molecular chaperones GRP78 and GRP94 (which would assist cells to resist cellular stress injury), and to determine the mRNA and protein expression levels of apoptosis promoting molecule Caspase-12 on the membrane of the ER, respectively. RESULTS: Protein expression levels of GRP78 and GRP94 were significantly increased in the TM-induced MSCs, and the mRNA level of Caspase-12 was also remarkably increased in the TM-induced MSCs (P<0.05). All these proved that the ERS model was successfully established by TM in MSC. Meanwhile, the mRNA and protein levels of GRP78 and GRP94 were all significantly increased compared with the model group (P<0.05 or P<0.01) after MSCs were treated with Tiantai No.1 while the mRNA and protein expression levels of Caspase-12 were significantly decreased compared with the model group (P<0.05 or P<0.01). This effect showed a dose dependent manner. CONCLUSION: Tiantai No.1 might attenuate the cell apoptosis induced by ERS injury, and thus protect the neurons against AD.

Effect of Tiantai No.1 on beta-amyloid-induced neurotoxicity and NF-kappa B and cAMP responsive element-binding protein.

OBJECTIVE: To investigate the effect and molecular mechanism of Tiantai No.1, a compound Chinese herbal preparation, for the prevention and reduction of neurotoxicity induced by beta-amyloid peptides (Abeta) in vitro and its effects on nuclear factor-kappa B (NF-kappa B) and cAMP responsive element-binding protein (CREB) pathways using the gene transfection technique. METHODS: B104 neuronal cells were used to examine the effects of Tiantai No.1 on lowering the neurotoxicity induced by Abeta. The cells were pre-treated with Tiantai No.1 at doses of 50, 100, 150, or 200 micro g/mL respectively for 3 days and co-treated with Tiantai No.1 and beta-amyloid peptide1-40 (A beta 1-40, 10 micro mol/L) for 48 h or post-treated with Tiantai No.1 for 48 h after the cells were exposed to beta-amyloid peptides25-35 (A beta 25-35) for 8 h. In gene transfection assays, cells were treated with Tiantai No.1 at 50 micro g/mL and 150 micro g/mL for 5 days or co-treated with Tiantai No.1 and A beta 1-40 (5 micro mo/L) for 3 days after electroporation for the evaluation of NF-kappa B and CREB expression. RESULTS: Pre-treating and co-treating B104 neuronal cells with Tiantai No.1 lowered the neurotoxicity induced by Abeta, and post-treating with Tiantai No.1 reduced or blocked B104 neuronal apoptotic death induced by Abeta (P<0.05, P<0.01). With a dose-dependent relationship, the same treatments increased the expression of NF-kappa B or CREB in B104 neuronal cells (P<0.05, P<0.01). Meanwhile, Tiantai No.1 reduced A beta -40 induced inhibition on NF-kappa B expression (P<0.01). CONCLUSIONS: Tiantai No.1 can protect neurons against the neurotoxicity induced by Abeta. The neuroprotective mechanisms may be associated with the activation of NF-kappa B and cAMP cellular signal pathways.