Electroacupuncture Promotes Angiogenesis in Mice with Cerebral Ischemia by Inhibiting miR-7.

OBJECTIVE: To observe the angiogenesis effect of electroacupuncture (EA) at Shuigou acupoint (GV 26) in the treatment of cerebral ischemia, and explore the value of miRNA-7 (miR-7) in it. METHODS: First, 48 mice were randomly divided into sham operation, middle cerebral artery occlusion (MCAO) model, and EA treatment groups. Then 9 mice were divided into carrier control group, miR-7 knockout group and miR-7 overexpression group (n=3 each group). Finally, 20 mice were divided into model and carrier control group, model and miR-7 knockout group, EA treatment and carrier control group and EA treatment and miR-7 overexpression group, with 3-6 mice in each group. The MCAO model was established in the MCAO and EA groups. Neurological deficit score and 2,3,5-triphenyltetrazolium chloride (TTC) staining were used to evaluate the severity of cerebral ischemia. Hematoxylin-eosin staining was used to describe basic pathological changes. Immunohistochemistry was used to quantify cerebral microvessel density. Real-time PCR and Western blot were used to detect the expression of miR-7 and its downstream target genes Krüppel-like factor 4/vascular endothelial growth factor (KLF4/VEGF) and angiopoietin-2 (ANG-2) in the ischemic cerebral cortex. RESULTS: After EA, neurological deficit scores and infarction volumes decreased, and the density of cerebral microvessels increased. In the MCAO group, miR-7 expression was higher than that in the sham group (P<0.01). After EA at GV 26, miR-7 expression decreased (P<0.01) and the expression of downstream target genes KLF4/VEGF and ANG-2 increased as compared with the MCAO group (P<0.01). After EA combined with overexpression of miR-7, the expression of downstream target genes KLF4/VEGF and ANG-2 decreased compared to the control EA group (P<0.01). After miR-7 knockdown, the expression of KLF4/VEGF and ANG-2 increased (P<0.05 or P<0.01). CONCLUSIONS: EA could promote angiogenesis in MCAO mice likely by inhibiting the expression of miR-7 and relieving inhibition of downstream target genes KLF4/VEGF and ANG-2.

Electroacupuncture ameliorates neuroinflammation by inhibiting TRPV4 channel in ischemic stroke.

AIMS: We investigated the potential mechanisms underlying the therapeutic efficacy of electroacupuncture (EA) at the Shuigou (GV26) and Baihui (GV20) acupoints in the treatment of ischemic stroke. METHODS: We assessed the therapeutic effects of EA on MCAO mice through behavioral studies and TTC staining. Various techniques, such as RT-PCR, immunofluorescence, and Western blots, were employed to evaluate the activation and polarization of microglia/macrophages, and changes in the TRPV4 ion channel. We used the TRPV4 antagonist GSK2193874 (GSK219) to verify the involvement of TRPV4 in the therapeutic effects of EA. RESULTS: EA effectively improved neurological impairments and reduced cerebral infarction volume in MCAO mice. It suppressed activated microglia/macrophages and inhibited their polarization toward the M1 phenotype post-MCAO. EA also downregulated the expression of pro-inflammatory cytokines, including Tnf-α, Il-6, Il-1ß, and Ccl-2 mRNA. Furthermore, EA reduced the elevated expression of TRPV4 following MCAO. Treatment with the TRPV4 antagonist GSK219 mirrored the effects of EA in MCAO mice. Notably, the combination of EA and GSK219 did not demonstrate an additive or synergistic effect. CONCLUSION: EA may inhibit neuroinflammation and exhibit a protective effect against ischemic brain injury by suppressing TRPV4 and the subsequent M1 polarization of microglia/macrophages.

The efficacy and safety of complementary and alternative medicine in the treatment of nausea and vomiting during pregnancy: A systematic review and meta-analysis.

Background: Complementary and alternative medicine (CAM) therapies are widely used for nausea and vomiting during pregnancy (NVP) due to the limitations of conventional medicine. However, their efficacy and safety remain controversial. Therefore, this meta-analysis was performed to assess the improvement of CAM therapy on NVP. Methods: Randomized controlled trials (RCTs) were searched for where the trial group was CAM and the control group was a conventional medicine or a placebo for NVP. This was done via 8 databases, including PubMed, EMBASE, the Cochrane Library, Web of Science, China National Knowledge Infrastructure, Wanfang, SinoMed, and VIP, from inception to October 25, 2022. The Grades of Recommendation, Assessment, Development and Evaluation (GRADE) was used to assess the quality of evidence. The Stata 15.0 software was used to perform the meta-analysis. Results: Thirty-three RCTs were included in this study. The acupuncture treatment was superior to conventional medicine at the effective rate [RR = 1.71, 95% CI (1.02, 2.86), P = 0.042; Low-quality evidence]. Ginger had more significant effects than conventional medicine at the Rhodes index [WMD = -0.52, 95% CI (-0.79, -0.24), P ≤ 0.001; Moderate-quality evidence] and it had the same effect as drugs to relieve vomiting [SMD = 0.30, 95% CI (-0.12, 0.73), P = 0.160; Low-quality evidence]. Compared with placebo, ginger had a higher effective rate [RR = 1.68, 95% CI (1.09, 2.57), P = 0.018; Low-quality evidence], and lower Visual analog scale (VAS) of Nausea [WMD = -1.21, 95% CI (-2.34, -0.08), P = 0.036; Low-quality evidence]. Ginger had the same antiemetic effect as placebo [WMD = 0.05, 95% CI (-0.23, 0.32), P = 0.743; Low-quality evidence]. Acupressure was superior to conventional medicine at the reduction of antiemetic drugs [SMD = -0.44, 95% CI (-0.77, -0.11), P = 0.008; Low-quality evidence], and at the effective rate [RR = 1.55, 95% CI (1.30, 1.86), P ≤ 0.001; Low-quality evidence]. Acupressure had the same effect as placebo at the effective rate [RR = 1.25, 95% CI (0.94, 1.65), P = 0.124; Low-quality evidence]. Overall, CAM therapy was safer than conventional medicine or a placebo. Conclusion: The results showed that CAM therapies were able to alleviate NVP. However, due to the low quality of existing RCTs, more RCTs with large sample sizes are needed to validate this conclusion in the future.

Electroacupuncture regulates microglia polarization via lncRNA-mediated hippo pathway after ischemic stroke.

Microglia polarization and microglia-mediated inflammation play a crucial role in the development of ischaemic brain injury. Electroacupuncture (EA) has the function of anti-inflammatory, which has been thoroughly validated and utilized to treat ischemic brain damage. The fundamental mechanism by which EA alleviates ischemic brain damage by decreasing microglia polarization and microglia-mediated inflammation, however, remains unknown. In the current study, the activation of microglia and inflammatory cytokines was analyzed to confirm the anti-inflammatory function of EA in middle cerebral artery occlusion (MCAO) rats. Whole-transcriptome sequencing was used to examine the differentially expressed lncRNAs in the control, MCAO, and MCAO +EA groups. Our findings demonstrated that EA treatment reduced microglia activation and inflammatory cytokine production. In addition, there are 44 lncRNAs were found significantly different in three groups, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of the predicted targets of these lncRNAs suggested that the Hippo pathway may contribute to the development of ischaemic brain injury and to the anti-inflammatory function of EA. Moreover, our data showed that lncRNA TCONS_00022826 (Lnc826) was upregulated in MCAO group, whereas blocked by EA treatment. Furthermore, in vitro OGD cell model data showed that Lnc826 promoted M1 polarization of microglia by regulating the Hippo pathway. Our data suggested that regulating microglia polarization via Lnc826-mediated hippo pathway is a possible mechanism of the EA treatment on ischemic brain injury.

Danshao Shugan Granule therapy for non-alcoholic fatty liver disease.

BACKGROUND: Danshao Shugan Granules (DSSG), a traditional Chinese medicine (TCM), is given to protect the liver. The objective is to evaluate the mechanisms of the effects of DSSG on non-alcoholic fatty liver disease (NAFLD). METHODS: 260 patients with NAFLD were randomly allocated to positive control drugs rosiglitazone (n = 30) and Silibinin (n = 50) as well as DSSG (n = 130) and combined DSSG/Silibinin (n = 50) groups, from which 90 patients in the DSSG group were further subdivided into 3 groups (n = 30, each) depending on the severity of symptoms. In total 33 Sprague-Dawley rats were assigned to normal (n = 10) or 45% high-fat diet (n = 23) groups, from which 9 rats served as negative controls, 10 as model controls and 10 were treated with DSSG. RESULTS: DSSG medications had significantly highest effects on B-ultrasonography finding improvements, and reductions of total cholesterol, triglyceride, aspartate transaminase and γ-glutamyl transpeptidase in NAFLD patients. Silibinin application only led to significantly highest alanine transaminase reductions and rosiglitazone medication to significantly highest fasting plasma glucose reductions. In a murine in vivo NAFLD model glucose (GLU), total cholesterol (TC) triacylglycerol (TG) as well as glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) and gamma-glutamyl transferase (GGT) serum concentrations were all significantly reduced (P < 0.001) and the expression of nuclear factor-κB (NF­κB) was significantly decreased in DSSG treated compared to untreated NAFLD animals (P < 0.001). In addition, the DSSG treated rats exhibited increased superoxide dismutase activity and reduced malondialdehyde values. CONCLUSIONS: DSSG was effective for treating NAFLD patients, which could be attributed to increased activity of superoxide dismutase, a decrease of malondialdehyde as well as reduced NF­κB activity in a NAFLD rat model.

Liuwei Dihuang Pill Attenuates Diabetic Nephropathy by Inhibiting Renal Fibrosis via TGF-ß/Smad2/3 Pathway.

Among all the complications of diabetes, diabetic nephropathy is a significant factor causing the end-stage renal disease associated with high death rates. Current treatment fails to produce an ideal outcome. Thus, searching for a new preventive drug is urgently needed. Liuwei Dihuang pill (LDP), a popular ancient Chinese medicine (TCM) prescription, has been applied to treat DN-like syndromes according to TCM theory. Here, we had established an animal model with DN and LDP therapy was put into use to assess its therapeutic effect in vivo. Our data showed that oxidative stress and TGF-ß/Smad2/3 pathway-induced renal fibrosis could be observed in the DN animal model. However, the treatment of LDP impeded the generation of ROS and attenuated renal fibrosis-related proteins in damaged kidneys through interference in the TGF-ß/Smad3 pathway. Our results indicated that LDP attenuated oxidative stress, accompanied by preventing the production of renal fibrosis through inhibiting the TGF-ß/Smad2/3 pathway.

Aquatic macrophytes mitigate the short-term negative effects of silver nanoparticles on denitrification and greenhouse gas emissions in riparian soils.

Silver nanoparticles (AgNPs) are increasingly released into the aquatic environments because of their extensive use in consumer products and industrial applications. Some researchers have explored the toxicity of AgNPs to nitrogen (N) and carbon (C) cycles, but little is known about the role of aquatic plants in regulating the impact of AgNPs on these biogeochemical processes and related microorganisms. Here, two 90-day pot experiments were conducted to determine the effect of AgNPs on denitrification rates and greenhouse gas emissions in riparian wetland soils, with or without emergent plants (Typha minima Funck). As a comparison, the toxicity of equal concentration of AgNO3 was also determined. The results showed that AgNPs released a great quantity of free Ag+, most of which was accumulated in soils, while little (less than 2%) was absorbed by plant shoots and roots. Both AgNPs and AgNO3 could increase the soil redox potential and affect the growth and nutrient (N and phosphorus) uptake of plants. In soils with plants, there was no significant difference in denitrification rates and emissions of N2O and CH4 between control and AgNPs or AgNO3 treatments at all tested concentrations (0.5, 1 and 10 mg kg-1). However, low levels of AgNPs (0.5 mg kg-1) significantly enhanced CO2 emission throughout the experiment. Interestingly, in the absence of plants, a high dosage (10 mg kg-1) of AgNPs generally inhibited soil denitrification and stimulated the emissions of CO2, CH4 and N2O in the short-term. Meanwhile, the abundance of key denitrifying genes (nirS and nirK) was significantly increased by exposure to 10 mg kg-1 AgNPs or AgNO3. Our results suggest that emergent plants can alleviate the short-term negative effects of AgNPs on N and C cycling processes in wetland soils through different pathways.

Electroacupuncture alleviates ischaemic brain injury by regulating the miRNA-34/Wnt/autophagy axis.

Electroacupuncture (EA), a modern form of acupuncture therapy, has been widely used for the treatment of ischaemic brain injury. However, the molecular mechanism by which EA improves ischaemic brain injury remains unclear. In the current study, middle cerebral artery occlusion (MCAO) rats were treated with EA. The infarct volumes and apoptosis of neurocytes were assessed to determine the therapeutic effect of EA. The differentially expressed miRNAs between the control, MCAO and MCAO treated with EA groups were detected by high-throughput sequencing. The results indicated that EA treatment decreased neurocyte apoptosis and ischaemic infarct volume. Between the three groups, miR-34, miR-235 and miR-275 were found to be significantly different. Furthermore, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway data suggest that the Wnt pathway may play an important role in ischaemic brain injury and in the treatment of EA. Our data documented that miR-34 was obviously increased in the MCAO group, while EA treatment decreased miR-34 expression. WNT1 was the target of miR-34 and was confirmed by a luciferase reporter assay. A previous study suggested that the Wnt pathway mediates autophagy in EA-pretreated MCAO mice. Our data further confirmed that EA treatment after MCAO also alleviated autophagy in the MCAO group. Our results suggest that EA treatment alleviates ischaemic brain injury by inhibiting autophagy through the miR-34/Wnt pathway.

Suppressing NK Cells by Astragaloside IV Protects Against Acute Ischemic Stroke in Mice Via Inhibiting STAT3.

Natural killer (NK) cells, a key member of innate lymphocytes, are a promising immunotherapeutic target for ischemic stroke. Astragaloside IV (ASIV) is isolated from Astragalus mongholicus Bunge (Fabaceae), a herbal medicine possessing immunomodulatory ability. This study investigated the effect of ASIV on NK cells during the acute stage of brain ischemic injury in a mouse model of middle cerebral artery occlusion (MCAO). MCAO mice treated with ASIV had better functional outcomes, smaller brain infarction and less NK cell brain infiltration. NK cell depletion echoed the protective effect of ASIV. Notably, ASIV did not enhance the protective effect of NK cell depletion against brain ischemic injury. ASIV inhibited glial cell-derived CCL2-mediated chemotaxis to prevent post-ischemic NK cell brain recruitment. Meanwhile, ASIV also abrogated NK cell-mediated cytolytic killing of neurons subjected to oxygen-glucose deprivation and suppressed NK cell-derived IFN-γ and NKG2D expression in the ischemic brain. The inhibitory effect of ASIV on NK cell brain infiltration and activation was mimicked by cryptotanshinone, a STAT3 inhibitor. There was no additive effect when ASIV and cryptotanshinone were used together. In conclusion, ASIV inhibits post-ischemic brain infiltration and activation of NK cells through STAT3 suppression, and this inhibitory effect of ASIV on NK cells plays a key role in its protection against acute ischemic brain injury. Our findings suggest that ASIV is a promising therapeutic candidate in NK cell-based immunotherapy for the treatment of acute ischemic stroke and pave the way for potential clinical trials.

Efficacy and Safety of Acupuncture at Tianshu (ST25) for Functional Constipation: Evidence from 10 Randomized Controlled Trials.

OBJECTIVE: To evaluate the evidence for the efficacy and safety of acupuncture at Tianshu (ST25) for functional constipation (FC). METHODS: We systematically searched seven databases to identify randomized controlled trials of acupuncture at ST25 alone or in combination with conventional therapy in the treatment of FC. Risk ratios (RRs) and mean differences (MDs) were calculated using RevMan 5.3 with 95% confidence interval (CI). RESULTS: The study included ten trials with 1568 participants. Meta-analysis showed that the Cleveland Constipation Score (CCS) for deep needling was significantly lower than that for lactulose (deep needling with low-frequency dilatational wave: MD -0.58, 95% CI -0.94 to -0.22; deep needling with sparse wave: MD -3.67, 95% CI -6.40 to -0.94; deep needling with high-frequency dilatational wave: MD -3.42, 95% CI -5.03 to -1.81). Furthermore, CCS for shallow needling with high-frequency dilatational wave was lower than that for lactulose (MD -1.77, 95% CI -3.40 to -0.14). In addition, when deep needling was combined with high-frequency dilatational wave, the weekly frequency of spontaneous defecation (FSD) was significantly higher than that for lactulose (MD 1.57, 95% CI 0.93 to 2.21). Colonic Transit Time (CTT) scores were significantly higher when deep needling was combined with sparse wave (MD -14.36, 95% CI -18.31 to -10.41) or high-frequency dilatational wave (MD -11.53, 95% CI -19.25 to -3.81). The time of first defecation after treatment (TFD) of the shallow needling therapy was significantly longer than that of the lactulose (MD 13.67, 95% CI 5.66 to 21.67). The CCS 6 months after treatment (CCS6m) for deep needling was significantly lower than that for lactulose (MD -4.90, 95% CI -5.97 to -3.84). Moreover, the FSD 6 months after treatment (FSD6m) for shallow needling was significantly higher than that for lactulose (MD 0.49, 95% CI 0.02 to 0.97). The adverse event (AE) rate for lactulose was significantly higher than that achieved with the needling treatments, and this held true for both deep needling therapy (RR 0.41, 95% CI 0.23 to 0.72) and shallow needling therapy (RR 0.33, 95% CI 0.15 to 0.77). CONCLUSIONS: The meta-analysis demonstrates that acupuncture at ST25 appears to be more effective than lactulose in the treatment of functional constipation. This was found to be especially true for deep needling with high-frequency dilatational wave, which had a greater impact on improving CCS, FSD, CTT, and CCS6m. Additionally, acupuncture at ST25 was shown to be safer than conventional treatment, with the rate of AE being significantly lower for both deep needling and shallow needling. The trial is registered with https://www.crd.york.ac.uk/prospero/(CRD42019141017)).

Wrist-ankle acupuncture and Fluoxetine in the treatment of post-stroke depression: a randomized controlled clinical trial.

OBJECTIVE: To investigate the effects of WAA combined with fluoxetine in the clinical treatment of post-stroke depression (PSD) . METHODS: In this randomized, controlled and single-blind trial, 105 PSD patients who met the inclusion criteria were randomly divided into three equal groups: Thin wrist-ankle acupuncture (WAA) group (Thin WAA needle + Fluoxetine), Thick WAA group (Thick WAA needle + Fluoxetine), and Sham WAA group (sham WAA needle + Fluoxetine). In this trial, the primary outcome was Hamilton Depression Scale (HAMD), while the secondary outcomes included Zung self-rating depression scale (SDS) and World Health Organization Quality of Life BREF (QQL). RESULTS: Ninety nine PSD patients completed all the treatment. The HAMD scores and SDS scores of all the three groups decreased after treatment (P < 0.05); thick WAA group and thin WAA group decreased more obviously than the sham WAA group (P < 0.05). There was no significant difference between the QQL scores of the three groups (P > 0.05). There was no significant difference in the scores of the three scales between the thick wrist ankle needles and the thin wrist ankle needles (P > 0.05). CONCLUSION: The present study showed that WAA combined with fluoxetine can relieve the symptoms of depression after stroke. WAA therapy could improve the antidepressant effect of fluoxetine.

Fire-needle acupuncture for upper limb spastic paralysis after stroke: Study protocol for a randomized controlled trial.

BACKGROUND: Fire-needle acupuncture, an important kind of acupuncture therapy, has been clinically used to treat upper limb spastic paralysis (ULSP) after stroke. Clinical experience has indicated that fire-needle acupuncture treatment takes less time, requires fewer visits, and has more rapid results and fewer side effects compared to chemical medicine alternatives. This study will evaluate the effects of fire-needle acupuncture for ULSP in the context of standardized clinical research and provide high-quality data to inform clinical procedures and future study design. METHODS/DESIGN: A randomized controlled trial will be carried out to evaluate the effects of fire-needle acupuncture therapy in patients with ULSP from stroke. ULSP patients (n = 120) will be recruited at Changhai Hospital in Shanghai, China. Patients will be randomly divided into three groups, including fire-needle acupuncture group (FAG), filiform-needle acupuncture group (FFAG) and rehabilitation treatment group (RTG). During the 3-week treatment, the FAG will be treated every two days, while FFAG and RTG will be treated 5 d in a row and then rest for 2 d. The Simplified Fugl-Meyer Motor Function Scale and Modified Ashworth Scale will be used as the primary outcome measures. Statistical analysis will be conducted by an independent statistician. DISCUSSION: Through this study, the utility of fire-needle acupuncture in treating ULSP after stroke will be tested, and some specific claims of fire-needle acupuncture therapy will be evaluated, such as relieving spasm and muscular tension, improving activities of daily living, rapidity of response and less frequency of treatment compared with other treatments. TRIAL REGISTRATION: Chinese Clinical Trial Registry (identifier: ChiCTR-IOR-17013875; registration date: 28 December 2016).

Application of FMEA and Bayesian Network in Risk Assessment of Fluidized Bed Granulation of Traditional Chinese Medicine / 中国实验方剂学杂志

Objective:To carry out the risk assessment on the factors in the process of granulation fluidized bed of traditional Chinese medicine(TCM) by using failure model and effect analysis(FMEA) and Bayesian network(BN), in order to effectively control risk factors and improve product quality. Method:The risk analysis of the fluidized bed granulation process was carried out by FMEA and the selected medium risk and high risk factors were taken as the main control points, the corresponding BN was established. The sensitivity analysis was used to screen out the main risk factors affecting particle fluidity, particle size uniformity, solubility and product cleanliness, the occurrence probability of each risk factor was determined by the evidence of unqualified particle quality, finally, taking fluidized bed granulation process of Sanye tablets as an example, the FMEA and BN were combined into the risk assessment process to verify the effectiveness and reliability of the method. Result:Based on the middle and high risk points of fluidized bed process, particle size of raw materials, moisture content and hygroscopicity of raw materials, dosage, concentration and addition amount of binder, cleaning degree and integrity of collection bag, and nozzle position, which were selected by FMEA, a fluidized bed granulation risk network with causality was constructed. Among them, hygroscopicity of raw materials, concentration and addition amount of binder, inlet temperature and atomization pressure were high probability risk factors, and the probability of occurrence were 55%, 63%, 59%and 58%, respectively. According to the Bayesian risk relationship network which controlled Sanye tablets fluidized bed granulation analysis results showed that the P values of inlet temperature, atomization pressure and concentration of binder were 0.003 4, 0.032 6 and 0.041 8, respectively in the regression model of influencing factors and particle size uniformity, indicating that there was a significant correlation between the three factors and the particle quality, which was basically consistent with the conclusion obtained by FMEA-BN method. Conclusion:The combination of FMEA and BN for visualized risk assessment of fluidized bed granulation helps to effectively control the risk factors in the granulation process, reduce product quality risks and provide strong support for the improvement of granulation process of TCM.

Wrist-ankle acupuncture (WAA) for primary dysmenorrhea (PD) of young females: study protocol for a randomized controlled trial.

BACKGROUND: Primary dysmenorrhea (PD) is one of the most common health complaints all over the world, specifically among young females. Acupuncture has been employed to relieve the pain-based symptoms and to avoid the side effects of conventional medication, and wrist-ankle acupuncture (WAA) has confirmed analgesic efficacy for various types of pain. The aim of this study is to evaluate the immediate analgesia effect of WAA on PD of young females. METHODS/DESIGN: This study will carry out a randomized parallel controlled single-blind trial to observe the immediate analgesia effect of WAA in PD of young females. Sixty participants who meet inclusion criteria will be recruited from September 2016 to September 2017 in Changhai hospital of China. They are randomly assigned to WAA therapy or sham acupuncture groups (30 patients for each group), and then receive real or sham acupuncture treatment, respectively. In this trial, the primary outcome measure is simple form of McGill pain questionnaire (SF-MPQ), while expectation and treatment credibility scale (ETCS), safety assessment, the COX menstrual symptom scale (CMSS), questionnaire about the feeling of being punctured are included in the secondary outcomes. DISCUSSION: This trial will be the first study protocol designed to evaluate the immediate analgesia effect of WAA in PD of young females. The strengths in methodology, including rigorous randomized, sham-controlled, participants-blinded and assessors-blinded, will guarantee the quality of this study. WAA doesn't require any needling sensation, so non-penetrating sham acupuncture can serve as an effective placebo intervention in this trial. TRIALS REGISTRATION: Chinese Clinical Trial Registry (identifier: ChiCTR-IOR-16008546 ; registration date: 27 May 2016).

Liuwei Dihuang pill treats diabetic nephropathy in rats by inhibiting of TGF-ß/SMADS, MAPK, and NF-kB and upregulating expression of cytoglobin in renal tissues.

Liuwei Dihuang pill (LDP) was assessed for its effects on renal deficiency.90 STZ induced DN rats were divided into groups (n = 22) without treatment (STZ) and LDP treated (STZ-L) (n = 23), Zhenwu decoction treated (STZ-Z) (n = 22), and valsartan treated (STZ-V) (n = 23) groups, with 16 normal control rats. Total urine protein (TP), blood urea nitrogen (BUN), and serum creatinine (Cr) were measured. Superoxide dismutase (SOD), nitric oxide synthase (NOS), and malondialdehyde (MDA) concentrations as well as expression/phosphorylation of SMAD3, SMAD2, and α-SMA, TGF-ß, RI /II, P38, ERK, and NF-kB in renal tissues were determined. In vitro experiments analyzed the effect of enhanced TGF-ß containing rat serums of the STZ groups on mesangial cells with and without transient transfection with a cytoglobin-containing plasmid.LDP treatment reduced the kidney coefficient, serum creatinine, blood urea nitrogen, and urine protein and prevented pathological changes. Expression of SOD and NOS in kidney tissue was increased but MDA expression reduced. LDP modulated multiple pathways, and its administration inhibited the phosphorylation of SMADS, ERK, p38, and the expression of NF-kB, α-SMA, and TGF-ß RI/II, and upregulated the expression of cytoglobin. In vitro studies revealed that overexpression of cytoglobin suppressed phosphorylation of Smad2, ERK, and p38 induced by TGF-ß and expression of NF-kB, α-SMA, and TGF-ß RI.LDP prevented renal fibrosis and protected glomerular mesangial cells by upregulation of cytoglobin and suppression of multiple pathways involving TGF-ß/SMADS, MAPK, NF-kB signaling.

Electroacupuncture Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulation of Autophagy and Apoptosis.

Background. The therapeutic mechanisms of cerebral ischemia treatment by acupuncture are yet not well addressed. Objective. We investigated the effects of electroacupuncture (EA) at GV26 observing the expression of autophagy-related proteins Beclin-1 and LC3B and proportion of apoptotic cells and Bcl-2 positive cells in MCAO/R model rats. Methods. Sprague-Dawley (SD) male rats were randomly assigned to 7 groups: model groups (M6h, M24h, and M72h), EA treatment groups (T6h, T24h, and T72h), and sham operation group (S). Neurological deficit and cerebral infarction volume were measured to assess the improvement effect, while the expression of Beclin-1 and LC3B and proportion of Tunel-positive and Bcl-2 positive cells were examined to explore EA effect on autophagy and apoptosis. Results. EA significantly decreased neurological deficit scores and the volume of cerebral infarction. Beclin-1 was significantly decreased in T24h, while LC3B-II/LC3B-I ratio markedly reduced in 6th hour. EA groups markedly reduced the number of Tunel positive cells, especially in T24h. Meanwhile, the number of Bcl-2 positive cells obviously increased after EA treatment, especially in T6h and T24h. Conclusions. The alleviation of inadequate autophagy and apoptosis may be a key mechanism involved in the reflex regulation of EA at GV26 to treat cerebral ischemia.

Comparison of Beneficial Effects of Acupuncture at PC6, LU7 & Non- acupuncture Points on Cardiac Cell Function in Myocardial Ischemia (MI) Rats by Testing Protein Expression of L-type Calcium Channel Protein and its Related Protein.

The mechanism of why electro-acupuncture (EA) at PC6 improves the heart function was investigated by studying how the L-type cardiac voltage-dependent calcium channel in myocardial ischemia (MI) is regulated. Cava,., Cavo and Cava2-61 are main component proteins of L-type calcium channel; CaM and Calmodulin kinase II (CaMKII) are Ca2+ channel associated proteins. In this experiment, MI was induced by injection of isoproterenol (ISO) in rats and electrocardiograms (ECGs) were recorded before and after every injection, and protein expressions of Cava,c, Cavp, Cava2_61, CaM and CaMKII were higher [The protein expression increased 43.39%, 54.85%, 47.08%, 48.29% and 50.36% respectively] than the control rats significantly (p<0.05). After MI induction, the MI rats were divided into three groups, including PC6 (Neiguan-point), LU7 (Lieque-point) and Non-acupuncture-point group, which were acupunctured once a day for 7 days respectively. After EA at PC6, the protein expressions showed obvious decrease [EA at PC6: the protein expressions of Cava1c, CavP, Cava2-81, CaM and CaMKII decreased 26.36%, 27.58%, 25.21%, 27.21% and 26.61% respectively.] and they are all lower than MI rats significantly (p<0.05). After EA at LU7 and Non-acupuncture-point, the protein expressions showed no significant changes. The effect of EA at PC6 was significantly better than LU7 and Non- acupuncture-point (p<0.05). PC6 is an acupoint of the pericardium meridian, and the pericardium meridian, which corresponds to opioid system according to Li P's research, can affect the cardio-vascular function directly. LU7 is located on the lung meridian; it cannot affect the heart function directly although the lung is related to the heart in blood circulation function so PC6 showed the target treating effect of meridian specificity on regulating the L-type calcium channel in MI.

[Apoptosis of hepatocellular induced by active ingredients of Zanthoxyli Radix by Raman spectroscopy].

The apoptosis of mono-hepatocellular induced by the active ingredients of the Zanthoxyli Radix was investigated using laser Raman spectroscopy. Hepatoma cells (BEL-7404) were treated with 10 mg•L⁻¹ nitidine chloride and 3 g•L⁻¹ the extracts of Zanthoxyli Radix, respectively, then were divided into two parts, one for fluorescence staining, the other for determination of Raman spectroscopy. The acquired spectra were then processed by background elimination, smoothing, and normalization. Fluorescence staining results showed that the nucleuses from untreated group were uniformly stained, while those from the group treated for 48 hours were densely stained and broken. The spectra results revealed that the intensity of peaks associated with nucleic acid and protein decreased after the cells were incubated with the extracts of Zanthoxyli Radix for 12, 24, 36 and 48 hours. The intensity of peaks at 785,1 002,1 175,1 660 cm⁻¹ was decreased with the time of the cells were incubated by the extracts of Zanthoxyli Radix. The results indicated that the extracts of Zanthoxyli Radix could induce the apoptosis of hepatoma cells and reduce the amount of nucleic acid and protein in the cells. There is a certain relevance between the drug treatment time and the efficacy. The above results suggest that Raman spectra can provide abundant information about the changes in biological macromolecules within the cells after incubated by the extracts of Zanthoxyli Radix and serve as an effective method for the real time measurement of apoptosis.

Traditional Chinese Acupuncture for Poststroke Depression: A Single-Blind Double-Simulated Randomized Controlled Trial.

OBJECTIVE: To evaluate the effectiveness and possible side-effect of treating poststroke depression patients by traditional Chinese body acupuncture. DESIGN: Single-blind double-simulated randomized controlled trial. SETTING: Inpatient wards of neurology and rehabilitation departments. PARTICIPANTS: Sixty-eight (68) participants who met the criteria were randomly assigned into two groups, 34 cases (32 completed) into intervention group and 34 cases (33 completed) into control group. INTERVENTIONS: Body acupuncture (Shuigou GV 26, Neiguan PC 6, and Zusanli ST 36) and oral placebo were used in intervention group while fluoxetine and minimal nontraditional acupuncture (minimally active penetrating) were used in control group. Patients in both groups were treated separately once a day for 6 weeks. OUTCOME MEASURES: Outcomes were measured using the 17-item Hamilton Depression Rating Scale (HAMD-17), and side-effects were measured using the Side Effect Rating Scale (SERS) of Asberg and a self-designed needling adverse events scale. Clinical effects of both groups were statistically valued before treatment, week-2, week-6, and month-3. RESULTS: The total curative effects of both groups are similar (p > 0.05; evaluated in week-6 and month-3), while intervention group had an earlier onset time at week-2 (p < 0.05). The intervention group has fewer side-effects in week-2 (p < 0.05). CONCLUSIONS: Body acupuncture was effective in reducing stroke patients' depressive symptoms and had fewer side-effects. It should be considered as an option for neuropsychiatric sequelae of stroke.

Wrist-ankle acupuncture (WAA) for precompetition nervous syndrome: study protocol for a randomized controlled trial.

BACKGROUND: Precompetition nervous syndrome comprises an excessive nervous and anxiety response to the high-pressure environment preceding a sporting competition. The use of acupuncture as a treatment option for anxiety, and wrist-ankle acupuncture (WAA) specifically in this instance, has been identified as a growing trend within the Western world. In our previous study, we have confirmed the efficacy of WAA for pre-examination anxiety. In this paper, we present a randomized controlled single-blind trial evaluating the use of WAA for precompetition nervous syndrome, comparing it with the intervention of sham acupuncture. METHODS/DESIGN: The study was designed as a randomized controlled single-blind trial to evaluate the effects of WAA for precompetition anxiety. The trial will be conducted in annual track and field events of Shanghai University of Sport. A total of 100 participants who meet inclusion criteria are randomly assigned by computerized randomization to receive WAA therapy or sham acupuncture. The group allocations and interventions are concealed to participants and statisticians. The Competition State Anxiety Scale (CSAI-2) is used as the primary outcome measure, while heart rate, blood pressure, respiratory frequency, tension syndrome curative effect evaluation and participants' feeling of acupuncture questionnaire are applied as secondary outcome measures. DISCUSSION: The results of this trial will confirm whether WAA is effective to treat precompetition anxiety in annual track and field events. TRIAL REGISTRATION: Chinese Clinical Trial Registry (identifier: ChiCTR-TRC-13003931; registration date: 22 October 2013).