Possible mechanisms of auricular acupoint stimulation in the treatment of migraine by activating auricular vagus nerve. / 从耳迷走神经探讨耳穴治疗偏头痛的可能机制.

Under the guidance of traditional Chinese medicine theory, the clinical research of auricular acupoint stimulation in the treatment of migraine has gained a lot, and the curative efficacy is definite, but its mechanism remains unclear. In the present paper, we discussed the efficacy of auricular acupoint stimulation including "transcutaneous auricular vagus nerve stimulation" (taVNS) in the treatment of migraine in recent years. Through bibliometric analysis, we screened out top 10 auricular acupoints (Shenmenï¼»TF4ï¼½, Pizhixiaï¼»AT4ï¼½, Jiaoganï¼»AH6aï¼½, Ganï¼»CO12ï¼½, Yidanï¼»CO11ï¼½, Neifenmiï¼»CO18ï¼½, Shenï¼»CO10ï¼½, Nieï¼»AT2ï¼½, Zhenï¼»AT3ï¼½ and Eï¼»AT1ï¼½) which were the most frequently used for migraine. Majority of these auricular acupoints just distributed in the region innervated by auricular vagus nerve. Thus, we thought that the analgesic effect of needling these auricular acupoints for migraine was produced by triggering the auricular vagus nerve, and concluded that the central mechanism underlying induction of analgesic effect by activating auricular vagus nerve may be achieved by activating the descending pain regulation pathway of the locus coeruleus nucleus and dorsal raphe nucleus. In addition, taVNS-induced 1) regulation of the activities of brain's default network and pain matrix, 2) activation of the cortical descending pain regulation pathway, and 3) inhibition of the neuroinflammatory response may also contribute to its ameliorating effect of migraine. This paper may provide ideas for the future research on the mechanism of auricular acupoint treatment of migraine.

Shrimp-waste based dispersant as oil spill treating agent: Biodegradation of dispersant and dispersed oil.

The emerging demand for the enhancement of biodegradation of persistent organic pollutants from marine oil spills using oil-treating agents to minimize the environmental impacts promotes the development of green dispersants. Shrimp waste is a potential raw material to generate green dispersants. The biodegradability of dispersed oil and dispersants themselves are key factors for the national consideration of the approval, stockpile, and usage of dispersants. However, it is unknown whether shrimp-waste-based dispersant (SWD) has high bioavailability or facilitates the biodegradation of dispersed oil. In this study, we tackled the biodegradation of oil dispersed by a purified SWD. Furthermore, the SWD biodegradability was evaluated by exploring the degradation genes via metagenomic sequencing, analyzing the enzymatic activities for dispersant biodegradation by molecular docking, and discussing the SWD toxicity. We discovered that the SWD facilitated the biodegradation of two crude oils (Alaska North Slope and Marine Fuel-No.6). The metagenomic analysis with molecular docking showed that fresh seawater had feasible enzymes to degrade the SWD to safety components. Additionally, the SWD was low toxic and high bioactive. The findings helped confirm that the purified SWD is an effective and eco-sustainable marine oil spill treating agent and tracked the biodegradation of dispersed oil and the SWD.

Recent advances in chemical and biological degradation of spilled oil: A review of dispersants application in the marine environment.

Growing concerns over the risk of accidental releases of oil into the marine environment have emphasized our need to improve both oil spill preparedness and response strategies. Among the available spill response options, dispersants offer the advantages of breaking oil slicks into small oil droplets and promoting their dilution, dissolution, and biodegradation within the water column. Thus dispersants can reduce the probability of oil slicks at sea from reaching coastal regions and reduce their direct impact on mammals, sea birds and shoreline ecosystems. To facilitate marine oil spill response operations, especially addressing spill incidents in remote/Arctic offshore regions, an in-depth understanding of the transportation, fate and effects of naturally/chemically dispersed oil is of great importance. This review provides a synthesis of recent research results studies related to the application of dispersants at the surface and in the deep sea, the fate and transportation of naturally and chemically dispersed oil, and dispersant application in the Arctic and ice-covered waters. Future perspectives have been provided to identify the research gaps and help industries and spill response organizations develop science-based guidelines and protocols for the application of dispersants application.

Metagenomic and Metatranscriptomic Responses of Chemical Dispersant Application during a Marine Dilbit Spill.

The global increase in marine transportation of dilbit (diluted bitumen) can increase the risk of spills, and the application of chemical dispersants remains a common response practice in spill events. To reliably evaluate dispersant effects on dilbit biodegradation over time, we set large-scale (1,500 mL) microcosms without nutrient addition using a low dilbit concentration (30 ppm). Shotgun metagenomics and metatranscriptomics were deployed to investigate microbial community responses to naturally and chemically dispersed dilbit. We found that the large-scale microcosms could produce more reproducible community trajectories than small-scale (250 mL) ones based on the 16S rRNA gene amplicon sequencing. In the early-stage large-scale microcosms, multiple genera were involved in the biodegradation of dilbit, while dispersant addition enriched primarily Alteromonas and competed for the utilization of dilbit, causing depressed degradation of aromatics. The metatranscriptomic-based metagenome-assembled genomes (MAG) further elucidated early-stage microbial antioxidation mechanism, which showed that dispersant addition triggered the increased expression of the antioxidation process genes of Alteromonas species. Differently, in the late stage, the microbial communities showed high diversity and richness and similar compositions and metabolic functions regardless of dispersant addition, indicating that the biotransformation of remaining compounds can occur within the post-oil communities. These findings can guide future microcosm studies and the application of chemical dispersants for responding to a marine dilbit spill. IMPORTANCE In this study, we employed microcosms to study the effects of marine dilbit spill and dispersant application on microbial community dynamics over time. We evaluated the impacts of microcosm scale and found that increasing the scale is beneficial for reducing community stochasticity, especially in the late stage of biodegradation. We observed that dispersant application suppressed aromatics biodegradation in the early stage (6 days), whereas exerting insignificant effects in the late stage (50 days), from both substance removal and metagenomic/metatranscriptomic perspectives. We further found that Alteromonas species are vital for the early-stage chemically dispersed oil biodegradation and clarified their degradation and antioxidation mechanisms. These findings help us to better understand microcosm studies and microbial roles for biodegrading dilbit and chemically dispersed dilbit and suggest that dispersant evaluation in large-scale systems and even through field trails would be more realistic after marine oil spill response.

Impacts of Frazil Ice on the Effectiveness of Oil Dispersion and Migration of Dispersed Oil.

Oil spills in the Arctic have drawn dramatic attention in recent years. Frazil ice, as the essential formation of sea ice, may affect the effectiveness of dispersants during oil spill response and the associated behaviors of dispersed oil. However, these impacts remain poorly understood, limiting the appropriate usage of dispersants in ice-covered regions. Herein this work explored the effects of frazil ice on the dispersion effectiveness of two dispersants (Corexit 9500A and hydrolyzed shrimp waste) and the migration of dispersed oil within frazil ice. We discovered that frazil ice inhibited dispersion effectiveness by attenuating water velocity. Permeable frazil ice encapsulated 11-30% of dispersed oil, implying a lower oil bioavailability. We thus proposed and verified a microscopic mechanism to unravel the migration of dispersed oil toward permeable constrictions in frazil ice. We predicted the concentration of dispersed oil encapsulated in frazil ice using bed filtration theory and verified the prediction through experiments. Furthermore, the presence of frazil ice can lead to the breakup and coalescence of dispersed oil. Overall, our findings would facilitate the appropriate planning and decision-making of dispersant-based oil spill response and a better understanding of the fate of dispersed oil in the frazil ice-infested ocean.

Fufang Xueshuantong alleviates diabetic retinopathy by activating the PPAR signalling pathway and complement and coagulation cascades.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Xueshuantong (FXST) is a traditional Chinese patent medicine composed of Panax notoginseng (Burkill) F.H.Chen (Araliaceae), Salvia miltiorrhiza Bunge (Lamiaceae), Astragalus propinquus Schischkin (Leguminosae), and Scrophularia ningpoensis Hemsl. (Scrophulariaceae). It has been widely used for the treatment of diabetic retinopathy (DR) and exerts a positive clinical therapeutic effect. AIM OF THE STUDY: The aim of this study was to observe the effect of FXST on diabetic rat retinas and investigate its pharmacological mechanism for improving DR. METHODS: The diabetic rat model was established by intraperitoneal injection of streptozotocin. The rats were divided into a normal group, diabetic group, and FXST group. The rats in the FXST group were treated with FXST by intragastric administration for 12 weeks while other rats were given the same volume of normal saline. The haemodynamic parameters of the central retinal artery in the rats were measured by ultrasound. Haematoxylin-eosin staining was utilised to observe the pathological structural changes in the retina. The apoptosis of retinal nerve cells was detected by terminal deoxynucleotidyl transferase dUTP nick end labelling. RNA sequencing was used to screen the differentially expressed genes (DEGs), and enrichment analyses were performed. The DEGs were validated through real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: The peak systolic velocity, end diastolic velocity, and mean velocity decreased while the resistance index and pulsatility index increased in the diabetic rat retinas. FXST also improved haemodynamics. In contrast with the diabetic group, FXST allayed the disorder and oedema of the retinal structure in addition to reversing the reductions in retinal thickness and retinal ganglion cell number. It also decreased the apoptosis index of retinal cells. A total of 1134 DEGs were identified by RNA sequencing in the FXST group compared to the diabetic group, including 814 upregulated genes and 320 downregulated genes. These genes were enriched in the complement and coagulation cascades as well as the peroxisome proliferator-activated receptor (PPAR) signalling pathway. Several DEGs, including PPAR gamma, perilipin 4, acyl-CoA dehydrogenase long chain, CD55 molecule, and plasminogen activator urokinase, were identified by qRT-PCR, and the results were consistent with the RNA sequencing data. CONCLUSIONS: FXST alleviates DR by improving the haemodynamics and morphological alterations of diabetic rat retinas, which are mediated by complement and coagulation cascades and the PPAR signalling pathway.

HuoXue JieDu formula improves diabetic retinopathy in rats by regulating microRNAs.

ETHNOPHARMACOLOGICAL RELEVANCE: HuoXue JieDu Formula (HXJDF) originates from classical formulas and was formed based on clinical experience. It is composed of Euonymus alatus (Thunb.) Siebold, Panax notoginseng (Burkill) F.H. Chen, the roots of Anguina kirilowii (Maxim.) Kuntze, and Coptis omeiensis (C. Chen) C.Y.Cheng. HXJDF prevents the deterioration of diabetic retinopathy. AIM OF THE STUDY: To evaluate the effects of HXJDF on diabetic retinopathy in rats and investigate the roles of miRNAs in the effects of HXJDF. MATERIALS AND METHODS: A single intraperitoneal injection of streptozotocin (STZ) (65 mg/kg) was used to induce diabetes in rats. Rats were divided into three groups: normal, diabetic, and diabetic + HXJDF. Rats were treated with HXJDF (15.4 g/kg) or water by oral gavage for twelve weeks. At the end of the treatment, rats were anaesthetized, and retinal haemodynamic changes were measured. Then, the retinas were removed and examined by haematoxylin and eosin (HE) staining and TUNEL assays. In addition, miRNA expression profiling was performed using miRNA microarrays and further validated by quantitative real-time PCR (qRT-PCR). RESULTS: Diabetes reduced peak systolic velocity (PSV), end-diastolic velocity (EDV), mean velocity (MV) and central retinal vein velocity (CRV) but increased the resistance index (RI) and pulsatility index (PI). In addition, in the diabetic group, retinal cell arrangement was disordered and loosely arranged, the retinal thickness and retinal ganglion cell (RGC) number decreased, and retinal cell apoptosis increased. In addition, 11 miRNAs were upregulated and 4 miRNAs were downregulated. After treatment, HXJDF improved retinal haemodynamics and morphologic changes, restored retinal thickness and RGC number and decreased retinal cell apoptosis. Furthermore, the changes in miRNA expression were significantly abolished by HXJDF. CONCLUSION: HXJDF may prevent DR by regulating the expression of miRNAs.

Microbial eco-physiological strategies for salinity-mediated crude oil biodegradation.

Salinity variability strongly affects the behaviors of oil degrading bacteria for spilled oil biodegradation in the marine environment. However, limited studies explored the strategies of microbes on salinity-mediated crude oil biodegradation. In this study, a halotolerant bio-emulsifier producer, Exiguobacterium sp. N41P, was examined as a model strain for Alaska North Slope (ANS) crude oil (0.5%, v/v) biodegradation. Results indicated that Exiguobacterium sp. N41P could tolerant a wide range of salinity (0-120 g/L NaCl) and achieve the highest degradation efficiency under the salinity of 15 g/L NaCl due to the highest biofilm formation ability. Moreover, increased salinity induced decreased cell surface hydrophobicity and a migration of microbial growth from oil phase to aqueous phase, leading to limited bio-emulsifier productivity and depressed degradation of insoluble long-chain n-alkanes while enhancing the degradation of relative soluble naphthalene. Research findings illustrated the microbial eco-physiological mechanism for spilled oil biodegradation under diverse salinities and advanced the understanding of sophisticated marine crude oil biodegradation process.

Differentiation of weathered chemically dispersed oil from weathered crude oil.

Oil fingerprinting is a crucial technology to trace the sources and behaviors of spilled oil. The use of dispersants enhances the stay of dispersed oil in a water column and changes the important properties of spilled oil. In case of fingerprinting of dispersed oil driven by dispersants, the fate and behaviors of biomarkers may be affected by the application of dispersants. Limited studies have investigated the statistical difference between fingerprinting of dispersed oil and non-dispersed oil using biomarkers, and the possible influence of the differences, if present. This study applied several principal component analyses (PCA) to differentiate weathered chemically dispersed oil from weathered crude (non-dispersed) oil using 103 diagnostic ratios of the same type of biomarkers and those of two types of biomarkers as input data. It showed that weathered chemically dispersed oil (CDO) can be differentiated from weathered crude oil (WCO) using specific diagnostic ratios that are affected by weathering. PCA analyses indicated the effects of the application of dispersants and weathering duration on weathering of biomarkers in CDO and WCO.

Generation of shrimp waste-based dispersant for oil spill response.

In this study, shrimp waste was enzymatically hydrolyzed to generate a green dispersant and the product was tested for crude oil dispersion in seawater. The hydrolysis process was first optimized based on the dispersant effectiveness (DE) of the product. The functional properties of the product were identified including stability, critical micelle concentration, and emulsification activity. Water was confirmed as a good solvent for dispersant generation when compared with three chemical solvents. The effects of salinity, mixing energy, and temperature on the dispersion of the Alaska North Slope (ANS) crude oil were examined. Microtox acute toxicity test was also conducted to evaluate the toxicity of the produced dispersant. In addition, DE of the product on three different types of crude oil, including ANS crude oil, Prudhoe Bay crude oil (PBC), and Arabian Light crude oil (ALC) was compared with that of the Corexit 9500, respectively. The research output could lead to a promising green solution to the oil spill problem and might result in many other environmental applications.

The Synthesis and Evaluation of Arctigenin Amino Acid Ester Derivatives.

The use of arctigenin (ARG), a traditional medicine with many pharmacological activities, has been restricted due to its poor solubility in water. Five amino acid derivatives of ARG have been synthesized using glycine, o-alanine, valine, leucine, and isoleucine, which have t-butyloxy carbonyl (BOC) as a protective group. In this study, we examined the effects of removing these protective groups. The results showed that the amino acid derivatives have better solubility and nitrite-clearing ability than ARG. Among the compounds tested, the amino acid derivatives without protective group were the best. Based on these results, ARG and its two amino acid derivatives without protective group (ARG8, ARG10) were selected to evaluate their anti-tumor activity in vivo at a dosage of 40 mg/kg. The results indicated that ARG8 and ARG10 both exhibit more anti-tumor activity than ARG in H22 tumor-bearing mice. The tumor inhibition rates of ARG8 and ARG10 were 69.27 and 43.58%, which was much higher than ARG. Furthermore, the mice treated with these compounds exhibited less damage to the liver, kidney and immune organs compared with the positive group. Furthermore, ARG8 and ARG10 improved the serum cytokine levels significantly compared to ARG. In brief, this study provides a method to improve the water solubility of drugs, and we also provide a reference basis for new drug development.

[Fast Detection of Camellia Sinensis Growth Process and Tea Quality Informations with Spectral Technology: A Review].

The research achievements and trends of spectral technology in fast detection of Camellia sinensis growth process information and tea quality information were being reviewed. Spectral technology is a kind of fast, nondestructive, efficient detection technology, which mainly contains infrared spectroscopy, fluorescence spectroscopy, Raman spectroscopy and mass spectroscopy. The rapid detection of Camellia sinensis growth process information and tea quality is helpful to realize the informatization and automation of tea production and ensure the tea quality and safety. This paper provides a review on its applications containing the detection of tea (Camellia sinensis) growing status(nitrogen, chlorophyll, diseases and insect pest), the discrimination of tea varieties, the grade discrimination of tea, the detection of tea internal quality (catechins, total polyphenols, caffeine, amino acid, pesticide residual and so on), the quality evaluation of tea beverage and tea by-product, the machinery of tea quality determination and discrimination. This paper briefly introduces the trends of the technology of the determination of tea growth process information, sensor and industrial application. In conclusion, spectral technology showed high potential to detect Camellia sinensis growth process information, to predict tea internal quality and to classify tea varieties and grades. Suitable chemometrics and preprocessing methods is helpful to improve the performance of the model and get rid of redundancy, which provides the possibility to develop the portable machinery. Future work is to develop the portable machinery and on-line detection system is recommended to improve the further application. The application and research achievement of spectral technology concerning about tea were outlined in this paper for the first time, which contained Camellia sinensis growth, tea production, the quality and safety of tea and by-produce and so on, as well as some problems to be solved and its future applicability in modern tea industrial.

[Electroacupuncture Combined with Clomiphene Promotes Pregnancy and Blastocyst Implantation Possibly by Up-regulating Expression of Insulin Receptor and Insulin Receptor Substrate 1 Proteins in Endometrium in Rats with PCOS].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention combined with clomiphene critate (CC) on the blastocyst implantation and pregnancy rate and expression of insulin receptor (INSR) and insulin receptor substrate 1 (IRS 1) proteins in the endometrium in rats with polycystic ovary syndrome (PCOS), so as to reveal its mechanisms underlying improvement of PCOS. METHODS: One hundred and twenty-five female SD rats were randomly divided into normal control, PCOS model, medication (CC), EA and EA+CC groups (n=25 in each group, 15 for checking blastocyst implantation, and 10 for Western blot). The PCOS model was established by subcutaneous injection of Dehydroepiandrosterone (DHEA) and fed with high-fat diet. Rats of the normal control group were treated by subcutaneous injection of sesame oil and fed with the normal forage. EA stimulation was applied to "Zhongwan" (CV 12), "Guanyuan" (CV 4) and bilateral "Tianshu" (ST 25) for 30 min, 3 times a week, 5 weeks altogether. Rats of the CC and EA+CC groups were fed with CC (100 mg·kg-1·d-1) for 2 days after regular restriction (30 min, 3 times a week, 5 weeks altogether). The pregnancy was determined by vaginal smear tests and the number of blastocyst implantation determined by examination of the uterus after execution. The expression of INSR and IRS 1 proteins in the endometrium was detected by Western blot. RESULTS: The pregnancy rate and the number of blastocyst implantation were significantly lower in the model group than in the normal control group (P<0.05), and remarkably increased after EA and EA+CC interventions (P<0.05). The effects of EA+CC were obviously superior to those of simple EA and simple medication in raising the pregnancy rate and the number of blastocyst implantation (P<0.05). No significant differences were found between the EA and CC groups in the pregnancy rate and the number of blastocyst implantation (P>0.05). The relative expression levels of both INSR and IRS 1 proteins were markedly lower in the model group than in the normal control group (P<0.05), and significantly higher in both EA and EA+CC groups than in the model group (P<0.05). No significant differences were found between the model and the CC groups and between the EA and EA+CC groups in the expression levels of INSR and IRS 1 proteins (P>0.05). CONCLUSIONS: EA intervention can improve pregnancy rate and the number of blastocyst implantation in PCOS rats, which may be related to its effects in up-regulating the expression of INSR and IRS 1 proteins in the endometrium.

[Effects of Electroacupuncture on mRNA Expressions of Insulin-receptor Substrates 1 and 2 in the Endometrium of PCOS Rats and Insulin Sensitivity].

Objective To observe the effects of electroacupuncture (EA) on gene expressions of insulin receptor substrate 1 (IRS1) and insulin receptor substrate 2 (IRS2) in the endometrium and insulin sensitivity (IS) of polycystic ovary syndrome (PCOS) rats, thereby providing basic evidence of clinical ap- plication of EA for improving the pregnancy rate of PCOS patients and reducing the abortion rate. Methods Dehydroepiandrosterone (DHEA) was subcutaneously injected to 24-day-old female SD rats to induce P- COS model. Besides, rats were fed with high-fat diet. Rats in the normal group were subcutaneously injected with sesame oil and fed with common forage. PCOS model rats were randomly divided into the model group and the EA group. All rats were intervened from 80 days old. Of them, EA was started to rats in the EA group, three times per week for 5 successive weeks. Rats in the normal group and the model group were only bound every day, but with no acupuncture, three times per week for 5 successive weeks. Blood was collected from caudal vein before and after treatment. Fasting blood glucose (FPG) was detected by oxidase method. Fasting insulin (FINS) level was determined by Roche electrochemical luminescence method. Homeostasis model assessment of insulin resistance ( HOMA-IR) index was also calculated. All rats were killed by decapitation by the end of intervention, and their endometria were collected. mRNA expressions of IRS1 and IRS2 in the endometrial tissue were detected by Real-time fluorescence quantitative PCR method. Results Pre-post-treatment changes of FPG level were not significantly different among the three groups (P >0. 05). Before treatment FINS level and HOMA-IR index were significantly higher in the model group than in the normal group (P <0. 05). After treatment, they were significantly lower in the EA group than in the model group (P <0. 05). Compared with the normal group, mRNA expressions of IRS1 and IRS2 in the endometrial tissue were decreased in the model group (P <0. 05). Compared with the model group, mRNA expressions of IRS1 and IRS2 in the endometrial tissue were increased in the EA group (P < 0. 05). Conclusion EA could improve IS and elevate mRNA expressions of IRS1 and IRS2 in the endome- trial tissue of PCOS rats, which might be one of mechanisms for improving endometrial IS.

Screening of biosurfactant-producing bacteria from offshore oil and gas platforms in North Atlantic Canada.

From offshore oil and gas platforms in North Atlantic Canada, crude oil, formation water, drilling mud, treated produced water and seawater samples were collected for screening potential biosurfactant producers. In total, 59 biosurfactant producers belong to 4 genera, namely, Bacillus, Rhodococcus, Halomonas, and Pseudomonas were identified and characterized. Phytogenetic trees based on 16S ribosomal deoxyribonucleic acid (16S rDNA) were constructed with isolated strains plus their closely related strains and isolated strains with biosurfactant producers in the literature, respectively. The distributions of the isolates were site and medium specific. The richness, diversity, and evenness of biosurfactant producer communities in oil and gas platform samples have been analyzed. Diverse isolates were found with featured properties such as effective reduction of surface tension, producing biosurfactants at high rate and stabilization of water-in-oil or oil-in-water emulsion. The producers and their corresponding biosurfactants had promising potential in applications such as offshore oil spill control, enhancing oil recovery and soil washing treatment of petroleum hydrocarbon-contaminated sites.

Hyperbaric oxygen treatment at various stages following chronic constriction injury produces different antinociceptive effects via regulation of P2X4R expression and apoptosis.

PURPOSE: The aims of this study were to investigate the effect of hyperbaric oxygen (HBO) treatment at various stages following chronic constriction injury (CCI) and to explore the underlying mechanisms of HBO treatment. METHODS: Forty adult male Sprague-Dawley rats were randomly assigned to five groups (n = 8 for each group): the sham group, CCI group, HBO1 group, HBO2 group, and HBO3 group. Neuropathic pain was induced by CCI of the sciatic nerve. HBO treatment began on postoperative days 1, 6, and 11 and continued for 5 days. The mechanical withdrawal threshold and thermal withdrawal latency were tested on preoperative day 3 and postoperative days 1, 3, 5, 7, 10, 14, and 21. The expression of P2X4R was determined by immunohistochemistry and western blot analysis. Cell apoptosis was measured using TUNEL staining. The expression of caspase 3 was measured using reverse transcription polymerase chain reaction (RT-PCR). Electron microscopy was used to determine the ultrastructural changes. RESULTS: Early HBO treatment beginning on postoperative day 1 produced a persistent antinociceptive effect and inhibited the CCI-induced increase in the expression of P2X4R without changing CCI-induced apoptosis. In contrast, late HBO treatment beginning on postoperative day 11 produced a persistent antinociceptive effect and inhibited CCI-induced apoptosis and upregulation of caspase-3 without changing the expression of P2X4R. In addition, late HBO treatment reduced CCI-induced ultrastructural damage. However, HBO treatment beginning on postoperative day 6 produced a transient antinociceptive effect without changing the expression of P2X4R or CCI-induced apoptosis. CONCLUSION: HBO treatment at various stages following CCI can produce antinociceptive effects via different mechanisms. Early HBO treatment is associated with inhibition of P2X4R expression, and late HBO treatment is associated with inhibition of cell apoptosis.

Huang-Lian-Jie-Du-Decotion induced protective autophagy against the injury of cerebral ischemia/reperfusion via MAPK-mTOR signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Huang-Lian-Jie-Du-Decotion (HLJDD, Hwangryun-Hae-Dok-Decotion in Japan), an ancient antipyretic and detoxifying traditional Chinese medicine formula, was reported to have protective effect on ischemic stroke. AIM OF THE RESEARCH: To investigate the therapeutic effect of HLJDD on ischemic stroke and explore its mode of action. MATERIAL AND METHODS: A model of ischemic stroke in the rat was established after transient middle cerebral artery occlusion (MCAO) followed by reperfusion. Rats were assigned randomly to groups of control, sham, transient ischemia/reperfusion (I/R), and three treatment groups by HLJDD at 2.5, 5.0, 10.0mg/kg. The neurological deficit, the cerebral infarct size, morphology abnormality, biochemical parameters were examined, and the levels of relevant proteins were determined by immunoblotting analysis to evaluate the protective effects of HLJDD on ischemic stroke and explore the underlying mechanism. RESULTS: Compared with I/R group, HLJDD significantly ameliorated neurological deficit and histopathology changes, decreased infarct area, and restored the levels of biochemical indicators including nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), glutathione disulfide (GSSG), total superoxide dismutase (T-SOD), Cu/Zn-SOD, Mn-SOD and glutathione peroxidase (GSH-PX). HLJDD also notably elevated the levels of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, and other autophagy related genes (Atgs), promoted the activation of extracellular signal-regulated kinases (ERK), protein kinase B (Akt), 3-phosphoinositide-dependent kinase (PDK1), and inhibited the activation of mammalian target of rapamycin (mTOR), c-Jun N-terminal protein kinases (JNK), p38, phosphatase and tensin homolog (PTEN). CONCLUSION: HLJDD showed neuroprotective effects on ischemic stroke, at least in part to the induced protective autophagy via the regulation of mitogen-activated protein kinase (MAPK) signals. This Akt-independent protective autophagy is favorable in the treatment of stroke, avoiding unfavorable side-effects associated with the inactivation of Akt. The efficacy of HLJDD on ischemic stroke and its safety warranted by its long-term clinical use in traditional Chinese medicine favored further study to develop HLJDD as an effective therapeutic agent to treat ischemic stroke.

[Effect of abdominal acupuncture therapy on the endocrine and metabolism in obesity-type polycystic ovarian syndrome patients].

OBJECTIVE: To observe the effect of abdominal acupuncture on the endocrine and metabolic level in obesity-type polycystic ovarian syndrome (PCOS) patients. METHODS: Eighty-six PCOS patients were randomly and equally divided into medication group and abdominal acupuncture group. Patients of medication group were treated with metformin (250 mg/time, t.i. d. in the 1st week, and 500 mg/time, t.i.d. thereafter) for 6 months,and those of abdominal acupuncture group were treated by abdominal acupuncture [Zhongwan (CV 12), Liangmen (ST 21), etc., once daily for 6 months]. Changes of the body height, body Mass index (BMI), waist-hip ratio (WHR), Ferriman-Galleey score (FGS), menstrual frequency (MF) and ovarian volume (OV) were determined. Serum luteotrophic hormone (LH), free testosterone (T), follicle-stimulating hormone (FSH) contents were detected with radioimmunoassay. Fasting blood glucose (FBG), fasting insulin (FIN), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), Homa insulin resistance index (IRI) were detected with chromatometry respectively. RESULTS: After the treatment, BMI, WHR, FGS and OV were reduced significantly in both medication and abdominal acupuncture groups (P < 0.05), while MF of the two groups increased evidently (P < 0.05), and the effects of abdominal acupuncture group were significantly superior to those of medication group in down-regulating BMI, WHR and upregulating MF (P < 0.05). Regarding the reproductive hormons, serum LH, LH/FSH and T levels in the two groups decreased significantly (P< 0.05), and the effect of abdominal acupuncture was superior to that of medication group in reducing serum T level (P < 0.05). Following the treatment, FBG, BG and FIN and INS contents 2 h after meal,and Homa IR in both medication and abdominal acupuncture groups all decreased considerably (P < 0.05), but without significant differences between them (P > 0.05). Regarding the blood lipid levels after the treatment, serum TC, TG, and LDL-C levels of the two groups decreased significantly (P < 0.05), while serum HDL-C level increased remarkably (P<0.05), without significant differences between the two groups in these indexes (P > 0.05). CONCLUSION: Abdominal acupuncture treatment can improve the endocrine and metabolic function of patients with obesity-type PCOS.