Schisandrin A alleviates renal fibrosis by inhibiting PKCß and oxidative stress.

BACKGROUND: Renal fibrosis is a common pathway that drives the advancement of numerous kidney maladies towards end-stage kidney disease (ESKD). Suppressing renal fibrosis holds paramount clinical importance in forestalling or retarding the transition of chronic kidney diseases (CKD) to renal failure. Schisandrin A (Sch A) possesses renoprotective effect in acute kidney injury (AKI), but its effects on renal fibrosis and underlying mechanism(s) have not been studied. STUDY DESIGN: Serum biochemical analysis, histological staining, and expression levels of related proteins were used to assess the effect of PKCß knockdown on renal fibrosis progression. Untargeted metabolomics was used to assess the effect of PKCß knockdown on serum metabolites. Unilateral Ureteral Obstruction (UUO) model and TGF-ß induced HK-2 cells and NIH-3T3 cells were used to evaluate the effect of Schisandrin A (Sch A) on renal fibrosis. PKCß overexpressed NIH-3T3 cells were used to verify the possible mechanism of Sch A. RESULTS: PKCß was upregulated in the UUO model. Knockdown of PKCß mitigated the progression of renal fibrosis by ameliorating perturbations in serum metabolites and curbing oxidative stress. Sch A alleviated renal fibrosis by downregulating the expression of PKCß in kidney. Treatment with Sch A significantly attenuated the upregulated proteins levels of FN, COL-I, PKCß, Vimentin and α-SMA in UUO mice. Moreover, Sch A exhibited a beneficial impact on markers associated with oxidative stress, including MDA, SOD, and GSH-Px. Overexpression of PKCß was found to counteract the renoprotective efficacy of Sch A in vitro. CONCLUSION: Sch A alleviates renal fibrosis by inhibiting PKCß and attenuating oxidative stress.

Safe utilization of bioresources in gentamicin mycelial residues by thermal treatment: Antibiotic degradation, resistance gene inactivation and available nutrients promotion.

Gentamicin mycelium residues (GMRs) abundant in organic substances were generated during the production of gentamicin. Inappropriate handling techniques not only waste valuable resources, they could also result in residual gentamicin into the natural environment, leading to the generation of antibiotic resistance genes (ARGs), which would cause a significant threat to ecological system and human health. In the present work, the effects of thermal treatment on the removal of residual gentamicin in GMRs, as well as the changes of associated ARGs abundance, antimicrobial activity and bioresources properties were investigated. The results indicated that the hazards of GMRs was significantly reduced through thermal treatment. The degradation rate of residual gentamicin in GMRs reached 100 %, the total abundance of gentamicin resistance genes declined from 8.20 to 1.14 × 10-5 and the antibacterial activity of the decomposition products of GMRs on Vibrio fischeri was markedly reduced at 200 °C for 120 min. Additionally, the thermal treatment remarkably influenced the bioresource properties of GMRs-decomposition products. The release of soluble organic matters including soluble carbohydrates and soluble proteins have been enhanced in GMRs, while excessively high temperatures could lead to a reduction of nutrient substances. Generally, thermal treatment technology was a promising strategy for synergistic reducing hazards and utilizing bioresources of GMRs.

Dihydroartemisinin attenuates ischemia/reperfusion-induced renal tubular senescence by activating autophagy.

Acute kidney injury (AKI) is an important factor for the occurrence and development of CKD. The protective effect of dihydroartemisinin on AKI and and reported mechanism have not been reported. In this study, we used two animal models including ischemia-reperfusion and UUO, as well as a high-glucose-stimulated HK-2 cell model, to evaluate the protective effect of dihydroartemisinin on premature senescence of renal tubular epithelial cells in vitro and in vivo. We demonstrated that dihydroartemisinin improved renal aging and renal injury by activating autophagy. In addition, we found that co-treatment with chloroquine, an autophagy inhibitor, abolished the anti-renal aging effect of dihydroartemisinin in vitro. These findings suggested that activation of autophagy/elimination of senescent cell might be a useful strategy to prevent AKI/UUO induced renal tubular senescence and fibrosis.

ß-Hydroxybutyric acid upregulated by Suhuang antitussive capsule ameliorates cough variant asthma through GSK3ß/AMPK-Nrf2 signal axis.

ETHNOPHARMACOLOGICAL RELEVANCE: Cough variant asthma (CVA) is a chronic inflammatory disease characterized by cough as the main symptom. Suhuang antitussive capsule (Suhuang), one of traditional Chinese patent medicines, mainly treats CVA clinically. Previous studies have shown that Suhuang significantly improved CVA, post-infectious cough (PIC), sputum obstruction and airway remodeling. However, the effect of Suhuang on ovalbumin-induced (OVA-induced) metabolic abnormalities in CVA is unknown. AIM OF THE STUDY: This study aimed to identify potential metabolites associated with efficacy of Suhuang in the treatment of CVA, and determined how Suhuang regulates metabolites, and differential metabolites reduce inflammation and oxidative stress. MATERIALS AND METHODS: Rats were given 1 mg OVA/100 mg aluminum hydroxide in the 1st and 7th days by intraperitoneal injection and challenged by atomizing inhalation of 1% OVA saline solution after two weeks to establish the CVA model. Rats were intragastrically (i.g.) administrated with Suhuang at 1.4 g/kg and ß-hydroxybutyric acid (ß-HB) were given with different concentrations (87.5 and 175 mg/kg/day) by intraperitoneal injection for 2 weeks. After 26 days, GC-MS-based metabolomic approach was applied to observe metabolic changes and search differential metabolites. The number of coughs, coughs latencies, enzyme-linked immunosorbent assay (ELISA), histological analysis and quantitative-polymerase chain reaction (Q-PCR) were used to investigate the effects of Suhuang. Then ß-HB on CVA rats, NLRP3 inflammasome and GSK3ß/AMPK/Nrf2 signalling pathway were detected by western blotting. RESULTS: The results showed that Suhuang treatment significantly enhanced the serum level of ß-HB. Interestingly, exposure to exogenous ß-HB was also protective against OVA-induced CVA. ß-HB significantly reduced the number of coughs and lengthened coughs latencies, improved lung injury, reduced the secretion of various cytokines, and directly inhibited the NLRP3 inflammasome. In addition, ß-HB increased the nuclear accumulation of Nrf2 by activating the GSK3ß/AMPK signaling axis, and then inactivating the NF-κB signaling pathway, effectively protecting OVA-induced CVA from oxidative stress and inflammation. CONCLUSIONS: The results of this study shows that ß-HB can reduce inflammation and oxidative stress, the increased production of ß-HB in serum might be the crucial factor for Suhuang to exert its effect in the treatment of CVA.

Insights into the effects of Zn exposure on the fate of tylosin resistance genes and dynamics of microbial community during co-composting with tylosin fermentation dregs and swine manure.

Though heavy metals are widely reported to induce antibiotic resistance propagation, how antibiotic resistance changes in response to heavy metal abundances remains unclearly. In this study, the tylosin fermentation dregs (TFDs) and swine manure co-composting process amended with two exposure levels of heavy metal Zn were performed. Results showed that the bioavailable Zn contents decreased 2.6-fold averagely, and the removal percentage of total tylosin resistance genes was around 23.5% after the co-composting completed. Furthermore, the tylosin resistance genes and some generic bacteria may exhibited a hormetic-like dose-response with the high-dosage inhibition and low dosage stimulation induced by bioavailable Zn contents during the co-composting process, which represented a beneficial aspect of adaptive responses to harmful environmental stimuli. This study provided a comprehensive understanding and predicted risk assessment for the Zn-contaminate solid wastes deposal and suggested that low levels of Zn or other heavy metals should receive more attention for their potential to the induction of resistance bacteria and propagation of antibiotic resistance genes.

The release of organic matter, nitrogen, phosphorus and heavy metals from erythromycin fermentation residue under heat-activated persulfate oxidation conditioning.

Erythromycin fermentation residue (EFR) is one kind of biological waste with high organic matter content. The recycling of EFR is not only beneficial to the environmental protection, but also to the economic development. In this study, the release of organic matter, nitrogen (N), phosphorus (P) and heavy metals (HMs) from EFR under heat-activated persulfate (PS) oxidation conditioning was investigated. Results indicated that oxidation conditioning promoted the release of soluble chemical oxygen demand (SCOD). Heat-activated PS oxidation conditioning boosted the release of total nitrogen (TN), ammonia­nitrogen (NH4+-N) and nitrate­nitrogen (NO3--N) into the supernatant, as well as the decomposition of organic nitrogen (ON). Concurrently, heat-activated PS oxidation conditioning facilitated the release of total phosphorus (TP), orthophosphate (PO43--P) and organic phosphorus (OP) into the supernatant, and the decomposition of OP. Furthermore, heat-activated PS oxidation conditioning resulted in the increase of release efficiencies of HMs. Therefore, heat-activated PS oxidation conditioning was beneficial to the release of organic matter, nutrients and HMs.

Biomechanical evaluation of calcium phosphate-based nanocomposite versus polymethylmethacrylate cement for percutaneous kyphoplasty.

BACKGROUND CONTEXT: Polymethylmethacrylate (PMMA) is the most commonly used filling material when performing percutaneous kyphoplasty (PKP) for the treatment of osteoporotic vertebral compression fractures. However, there are some inherent and unavoidable drawbacks with the clinical use of PMMA. PMMA bone cement tends to leak during injection, which can lead to injury of the spinal nerves and spinal cord. Moreover, the mechanical strength of PMMA-augmented vertebral bodies is extraordinary and this high level of mechanical strength might predispose to adjacent vertebral fractures. A novel biodegradable calcium phosphate-based nanocomposite (CPN) for PKP augmentation has recently been developed to potentially avoid these issues. PURPOSE: By comparison with PMMA, the leakage characteristics, biomechanical properties, and dispersion of CPN were evaluated when used for PKP. STUDY DESIGN: Biomechanical evaluation and studies on the dispersion and anti-leakage properties of CPN and PMMA cements were performed and compared using cadaveric vertebral fracture model, sheep vertebral fracture model, and simulated rigid foam model. METHODS: Sheep vertebral bodies were decalcified by ethylenediaminetetraacetic acid disodium salt (EDTA-Na2) to simulate osteoporosis in vitro. After compression to create wedge-shaped fractures using a self-designed fracture creation tool, human cadaveric vertebrae and decalcified sheep vertebrae were augmented by PKP. In addition, three L5 vertebral bodies from human cadavers were used in a contrast vertebroplasty (VP) augmentation experiment. Occurrence of cement leakage was observed and compared between CPN and PMMA during the process of vertebral augmentation. Open-cell rigid foam model (Sawbones#1522-507) was used to create a simulated leakage model for the evaluation of the leakage characteristics of CPN and PMMA with different viscosities. The augmentation effects of CPN and PMMA were evaluated in human cadaveric and decalcified sheep vertebral models and then compared to the results from solid rigid foam model (Sawbones#1522-23). The dispersion abilities of CPN and PMMA were evaluated via three methods as follows. The dispersion volume and dispersion ratio were calculated by three-dimensional reconstruction using human vertebral body CT scans; the ratio of cement area to injection volume was calculated from three-dimensional sections of micro-CT scans of a sheep vertebra; and the micro-CT images of cement dispersion in open-cell rigid foam model (Sawbones#1522-507) were compared between CPN and PMMA. This study was funded by the National Natural Science Foundation of China (No. 81622032, 190,000 dollars and No. 51672184, 90,600 dollars), Principal Project of Natural Science Research of Jiangsu Higher Education Institutions (No. 17KJA180011, 22,000 dollars), and Jiangsu Innovation and Entrepreneurship Program (146,000 dollars). RESULTS: There was no significant difference in vertebral height between CPN and PMMA during PKP augmentation and both cements restored the vertebral height after augmentation. In PKP augmentation experiment, posterior wall cement leakage occurred in 75% of human vertebrae augmented with PMMA; however, no leakage occurred in human vertebrae augmented with CPN. Anterior leakage occurred in all vertebrae augmented by PMMA, while in only 75% of vertebra augmented by CPN. Furthermore, CPN and PMMA had completely different leakage patterns in the simulated rigid foam model whether administered at the same injection speed or under the same injection force, suggesting that CPN has anti-leakage characteristics. The augmentation in human cadaveric vertebrae was lower with CPN compared to PMMA (1,668±816 N vs. 2,212±813 N, p=.459, respectively), but this difference was not significant. The augmentation force in sheep vertebral bodies reached 1,393±433 N when augmented with PMMA, but 1,108±284 N when augmented with CPN. The dispersion of CPN was better, and the dispersion volume and ratio were greater, with CPN than with PMMA. Imaging of the open-cell rigid foam model showed completely different dispersion modes for CPN and PMMA. After injection, the PMMA cement formed a contracted clump in the open-cell rigid foam model. However, the CPN cement extended many antennae outward, appearing to spread to the surrounding area. The surface areas of the CPN cement blocks with different liquid-to-solid ratios were significantly larger than the surface area of the PMMA cement in the open-cell rigid foam model (p<.05). CONCLUSIONS: CPN has anti-leakage properties, which might be related to its high viscosity and viscoplasticity. CPN had a slightly lower augmentation force than PMMA when used in cadaveric vertebrae, decalcified sheep vertebrae, and in the standard rigid foam model. However, CPN diffused more easily into cancellous bone than did PMMA and encapsulated bone tissue during the dispersion process. The excellent dispersion of CPN generated better interdigitation with cancellous bone, which may be why the augmentation effect of CPN is similar to that of PMMA. CLINICAL SIGNIFICANCE: Biodegradable CPN is a potential alternative to PMMA cement in PKP surgery, in which CPN is likely to reduce the cement leakage during the surgery and avoid the post-surgery complications caused by excessive strengths and nondegradability of PMMA cement.

ß-Arrestin1 alleviates acute pancreatitis via repression of NF-κBp65 activation.

BACKGROUND AND AIM: ß-Arrestins (ß-arrs) are regulators and mediators of G protein-coupled receptor signaling that are functionally involved in inflammation. Nuclear factor-κB p65 (NF-κBp65) activation has been observed early in the onset of pancreatitis. However, the effect of ß-arrs in acute pancreatitis (AP) is unclear. The aim of this study is to investigate whether ß-arrs are involved in AP through activation of NF-κBp65. METHODS: Acute pancreatitis was induced by either caerulein injection or choline-deficient supplemented with ethionine diet (CDE). ß-arr1 wild-type and ß-arr1 knockout mice were used in the experiment. The survival rate was calculated in the CDE model mice. Histological and western blot analyses were performed in the caerulein model. Inflammatory mediators were detected by real-time polymerase chain reaction in the caerulein-induced AP mice. Furthermore, AR42J and PANC-1 cell lines were used to further study the effects of ß-arr1 in caerulein-induced pancreatic cells. RESULTS: ß-Arr1 but not ß-arr2 is significantly downregulated in caerulein-induced AP in mice. Targeted deletion of ß-arr1 notably upregulated expression of the pancreatic inflammatory mediators including tumor necrosis factor α and interleukin 1ß as well as interleukin 6 and aggravated AP in caerulein-induced mice. ß-Arr1 deficiency increased mortality in mice with CDE-induced AP. Further, ß-arr1 deficiency enhanced caerulein-induced phosphorylation of NF-κBp65 both in vivo and in vitro. CONCLUSION: ß-Arr1 alleviates AP via repression of NF-κBp65 activation, and it is a potentially therapeutic target for AP.

Bis(pyrene)-Doped Cationic Dipeptide Nanoparticles for Two-Photon-Activated Photodynamic Therapy.

At present, one of main problems for photodynamic therapy (PDT) is how to improve the treatment depth. Two-photon activated (TPA) developed recently provide a possible solution for it. In this work, we report the energy-transferring assembled cationic dipeptide nanoparticles for two-photon activated photodynamic therapy (TPA-PDT). In the nanoparticles, the coencapsulated two-photon fluorescent dye bis(pyrene) (BP) is an energy donor, and a photosensitizer rose bengal (RB) is an acceptor based on an intraparticle fluorescence resonance energy transfer (FRET) mechanism. BP in the nanoparticles can be excited by one- or two- photon laser. And then, the energy of BP was transferred to RB, which highly enhanced the generation of singlet oxygen. The cellular experiments indicated that this nanosystem can induce the cytotoxicity under one- and two-photon irradiation, which allows further applications of FRET-based biomaterials for TPA-PDT.

Identification of a flavonoid C-glycoside as potent antioxidant.

Flavonoids have been documented to have good antioxidant activities in vitro. However, reports on the cellular antioxidant activities of flavonoid C-glycosides are very limited. In this work, an apigenin C-glycoside was purified from Artocarpus heterophyllus by column chromatography and was identified to be 2″-O-ß-D-xylosylvitexin by nuclear magnetic resonance spectroscopy. The cellular antioxidant activity and anticancer activity of 2″-O-ß-D-xylosylvitexin were evaluated for the first time. The quantitative structure-activity relationship was analysed by molecular modeling. Apigenin presented an unexpected cellular antioxidation behaviour. It had an antioxidant activity at low concentration and a prooxidant activity at high concentration, whereas 2″-O-ß-D-xylosylvitexin showed a dose-dependent cellular antioxidant activity. It indicated that C-glycosidation improved the cellular antioxidation performance of apigenin and eliminated the prooxidant effect. The ortho-dihydroxyl at C-3'/C-4' and C-3 hydroxyl in the flavonoid skeleton play important roles in the antioxidation behaviour. The cell proliferation assay revealed a low cytotoxicity of 2″-O-ß-D-xylosylvitexin.

Effect of dry mycelium of Penicillium chrysogenum fertilizer on soil microbial community composition, enzyme activities and snap bean growth.

The dry mycelium fertilizer (DMF) was produced from penicillin fermentation fungi mycelium (PFFM) following an acid-heating pretreatment to degrade the residual penicillin. In this study, it was applied into soil as fertilizer to investigate its effects on soil properties, phytotoxicity, microbial community composition, enzyme activities, and growth of snap bean in greenhouse. As the results show, pH, total nitrogen, total phosphorus, total potassium, and organic matter of soil with DMF treatments were generally higher than CON treatment. In addition, the applied DMF did not cause heavy metal and residual drug pollution of the modified soil. The lowest GI values (<0.3) were recorded at DMF8 (36 kg DMF/plat) on the first days after applying the fertilizer, indicating that severe phytotoxicity appeared in the DMF8-modified soil. Results of microbial population and enzyme activities illustrated that DMF was rapidly decomposed and the decomposition process significantly affected microbial growth and enzyme activities. The DMF-modified soil phytotoxicity decreased at the late fertilization time. DMF1 was considered as the optimum amount of DMF dose based on principal component analysis scores. Plant height and plant yield of snap bean were remarkably enhanced with the optimum DMF dose.

Analysis of medicine consumption in peacekeeping level II hospitals.

BACKGROUND: The peacekeeping military units of contributing countries are unfamiliar with the conditions prevailing in foreign mission areas and therefore have difficulties with medical supplies storage. AIM: The aim of this study is to provide reasonable and practical guidance on the maintenance of medical supplies in the peacekeeping military units of contributing countries. MATERIALS AND METHODS: A total of 1,972 prescriptions were received by the pharmacy in the peacekeeping level II hospital in the Republic of Sudan from February to July of 2009 including a total of 186 drug categories and 17,713 minimum packing units. Pairwise comparison was performed using the c2 test. When the total number of samples was smaller than 40, the Fisher's exact test was adopted for pairwise comparison. RESULTS: The majority of the consumed medicines mainly belonged to 6 categories, including specialty drugs, anti-microbial drugs, Chinese patent medicines, gastrointestinal drugs, central nervous system drugs, and drugs regulating fluids, electrolytes, and acid-base balance. Altogether, the drugs in the 6 categories accounted for 74% of all consumed medicines that were divided into a total of 20 categories. CONCLUSIONS: Medicine consumption in peacekeeping level II hospitals is unique, therefore the drugs used in military medical facilities should be prepared according to their actual needs in the area of peacekeeping operations.

[Multicentral randomized controlled study on effects of acupuncture at Zusanli (ST 36) and Xuanzhong (GB 39) on cerebrovascular function in the patient of ischemic stroke].

OBJECTIVE: To explore effect of acupuncture at Zusanli (ST 36) and Xuanzhong (GB 39) on cerebrovascular function in the patient of ischemic stroke. METHODS: Three central, single blind, randomized controlled trial method was adopted, and 160 cases were randomly divided into an acupuncture group and a control group, 80 cases in each group. The two groups were treated by routine treatment for ischemic stroke with acupuncture at Zusanli (ST 36) and Xuanzhong (GB 39) added in the acupuncture group. Changes of TCD cerebrovascular blood flow indexes before and after treatment were evaluated. RESULTS: After treatment, TCD indexes significantly improved in the acupuncture group (P < 0.05, P < 0.01) with a significant difference as compared with that in the control group (P < 0.05). CONCLUSION: Acupuncture at Zusanli (ST 36) and Xuanzhong (GB 39) can significantly improve cerebral vasomotoricity, cerebral blood flow auto-regulative function, cerebral hemisphere collateral circulation comprehental function in the patient of ischemic stroke.

Immunoaffinity column cleanup with liquid chromatography using post-column bromination for aflatoxins in medicinal herbs and plant extracts.

A new and accurate method to quantitate aflatoxins in medicinal herbs is developed. This method consists of extraction of the sample with MeOH-H2O (70:30) followed by clean-up of the extracts with immunoaffinity columns and, finally, high-performance liquid chromatographic determination with fluorescence detection. Aflatoxins B1 and G1 are determined as their bromine derivatives, produced in an online post-column derivatization system. The overall average recoveries for three different medicinal herbs spiked at levels of 1.3 and 2.6 ng/g of total aflatoxins range from 93% to 97%. The detection limit is 0.15 ng/g for both G2 and B2 and 0.20 ng/g for both G1 and B1, based on a signal-to-noise ratio of 3:1 and a precision (within-laboratory relative standard deviation) ranging from 0.8% to 1.4%. The use of immunoaffinity columns provides excellent clean-up of these particular extracts, which are generally difficult to analyze. The method is applied successfully to 96 samples of natural drugs.

[Influences of repeated lower +Gz exposures on high +Gz exposure induced brain injury in rats].

Objective. To explore the influence of repeated lower +Gz exposures on high +Gz exposure induced brain injury in rats. Method. Forty SD male rats were randomly divided into control group (5 rats), +10 Gz/5 min group (5 rats) +4 Gz exposure one time, three times and five times group (10 rats each group). After 1 d or 6 d of +4 Gz exposure each group were exposed to +10 Gz again. Three days after +10 Gz exposure the neuron damage was observed by light microscope in HE stained section. Result. There was no brain damage after repeated +4 Gz/3 min exposure 5 times so it was reasonable to use this exposure intensity as ischemia stimulation. +10 Gz/5 min exposure could result in irreversible neuron damage such as neuron degeneration and coagulation necrosis. The experiment results suggested that after +10 Gz/5 min exposure there were degenerated neurons in cortex, hippocampus and thalamus. The number of degenerated neurons were obviously decreased in cortex, hippocampus and thalamus when exposed to +10 Gz/5 min again after repeated +4 Gz/3 min 3-5 times. Conclusion. The degree of neuron damage was obviously slight at the time of exposure to +10 Gz/5 min again after repeated +4 Gz/3 min 3-5 times. The ischemia tolerance at the time of exposure to +Gz was similar to other brain ischemia.

Immunocytochemical localization of substance P receptor in hypothalamic oxytocin-containing neurons of C57 mice.

With the use of double immunofluorescence, we have examined the distribution of oxytocin-containing neurons that express substance P receptor (SPR) in the hypothalamus of C57 mice. The distribution of oxytocin-like immunoreactive neurons overlapped with that of SPR-like immunoreactive neurons in the paraventricular nucleus and supraoptic nucleus of the hypothalamus. Neurons showing both oxytocin- and SPR-like immunoreactivities were predominantly found in both nuclei. A few neurons that were double-labeled with oxytocin- and SPR-like immunoreactivities were also scattered in the hypothalamic periventricular and preoptic regions. Semi-quantitative analysis indicated that about 94% of the oxytocin-like neurons displayed SPR-like immunoreactivity. These double-labeled cells constituted about 91% of the SPR-like neurons in the aforementioned regions. The present study provides morphological evidence for tachykinin-induced modulation of oxytocin-containing neurons as mediated by substance P receptor in the hypothalamus of mammals.

[Study of the acupuncture effect on monoamine transmitters in rabbit plasma and brain tissue by high performance liquid chromatography with electrochemical detection].

A rapid and simple method for the study of the acupuncture effect on monoamine transmitters and related compounds in rabbit plasma and brain tissue by high performance liquid chromatography with electrochemical detection was developed. An ODS column was selected as the separation column at 25 degrees C, and pH 4.50, 0.02 mol/L of trisodium citrate-0.05 mol/L sodium phosphate dibasic to methanol (95:5, volume ratio) without ion-pair at a flow rate of 1.0 mL/min. Four compounds, epinephrine (E), norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT), were simultaneously separated and determined under the above conditions. Twenty rabbits were investigated after the acupuncture action upon the central neurotransmitters. The sufficient data showed that acupuncture could significantly affect the activities of the neurotransmitters including E, NE, DA and 5-HT, and the changed functions of the neurotransmitter systems induced by acupuncture not only lead to the neurotransmitter content increase both in brain and plasma but also cause the increase of rabbit breed ability. The results show that the method is very simple and fast. The method is valuable not only for clinical diagnosis but also for research work.