Effects of electroacupuncture on rats with cognitive impairment: An iTRAQ-based proteomics analysis / 中西医结合学报

OBJECTIVE@#The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment.@*METHODS@#Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ.@*RESULTS@#Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein β1 (Hspb1) was the highest in the EA group compared to the model group.@*CONCLUSION@#EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.

Potentiating effect and mechanism of extract of Jingfang Granules on activation of macrophages / 中国中药杂志

This study aimed to explore the potentiating effect and mechanism of the extract of Jingfang Granules(JFG) on the activation of macrophages. The RAW264.7 cells were treated with JFG extract and then stimulated by multiple agents. Subsequently, mRNA was extracted, and reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the mRNA transcription of multiple cytokines in RAW264.7 cells. The levels of cytokines in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA). In addition, the intracellular proteins were extracted and the activation of signaling pathways was determined by Western blot. The results showed that JFG extract alone could not promote or slightly promote the mRNA transcription of TNF-α, IL-6, IL-1β, MIP-1α, MCP-1, CCL5, IP-10, and IFN-β, and significantly enhance the mRNA transcription of these cytokines in RAW264.7 cells induced by R848 and CpG in a dose-dependent manner. Furthermore, JFG extract also potentiated the secretion of TNF-α, IL-6, MCP-1, and IFN-β by RAW264.7 cells stimulated with R848 and CpG. As revealed by mechanism analysis, JFG extract enhanced the phosphorylation of p38, ERK1/2, IRF3, STAT1, and STAT3 in RAW264.7 cells induced by CpG. The findings of this study indicate that JFG extract can selectively potentiate the activation of macrophages induced by R848 and CpG, which may be attributed to the promotion of the activation of MAPKs, IRF3, and STAT1/3 signaling pathways.

Condensed tannins from roots of Indigofera stachyodes / 中国中药杂志

Eleven condensed tannins were isolated from the roots of Indigofera stachyodes by various column chromatography techniques including silica gel, octadecyl silica(ODS), Sephadex LH-20, and semi-preparative high performance liquid chromatography(HPLC). These compounds were identified on the basis of physicochemical properties, nuclear magnetic resonance(NMR) and mass spectrometry(MS) data as stachyotannin A(1), epicatechin-(2β→O→7,4β→8)-epiafzelechin-(4β→8)-catechin(2), cinnamtannin D1(3), cinnamtannin B1(4), epicatechin-(2β→O→7,4β→8)-epiafzelechin-(4α→8)-epicatechin(5), gambiriin C(6), proanthocyanidin A1(7), proanthocyanidin A2(8), aesculitannin B(9), proanthocyanidin A4(10), and procyanidin B5(11). Compound 1 is a new compound. Compounds 2-11 were isolated from Indigofera for the first time. Furthermore, compounds 1, 2, and 4-11 showed inhibitory effects on thrombin-induced ATP release in platelets.

Synthesis and bioactivities of phenazine-1-carboxylic piperazine derivatives.

Phenazine-1-carboxylic acid (PCA) as a natural product which has significant inhibition effects against many soil-borne fungal phytopathogens in agricultural application and has been registered in China as the fungicide against rice sheath blight. In order to find new higher fungicidal activities lead compounds and develop new eco-friendly agrochemicals, we introduced substructure piperazines which also have high biological activity into PCA, designed and synthesized a series of phenazine-1-carboxylic piperazine derivatives, and their structures were confirmed by 1H NMR and HRMS. Most compounds exhibited certain in vitro fungicidal activities. In particular, Compounds 5r exhibited the activity against all the tested pathogenic fungi, such as Rhizoctonia solani, Alternaria solani, Fusarium oxysporum, Fusarium graminearum, Pyricularia oryzac Cavgra, with the EC50 value of 24.6µM, 42.9µM, 73.7µM, 73.8µM, 34.2µM, respectively, more potent activities than PCA (33.2µM, 81.5µM, 186.5µM, 176.4µM, 37.3µM). This result provided a highly active lead compound for the further structure optimization design.

Rapid discovery and identification of the anti-inflammatory constituents in Zhi-Shi-Zhi-Zi-Chi-Tang / 中国天然药物

The anti-inflammatory active ingredients of Zhi-Shi-Zhi-Zi-Chi-Tang (ZSZZCT), a traditional Chinese medicine formula, were predicted and identified using an approach based on activity index, LC-MS, semi-preparative LC and NMR. Firstly, the whole extract of ZSZZCT was analyzed using liquid chromatography-quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) and liquid chromatography - ion trap mass spectrometry (LC-IT-MS), 79 constituents were detected and 39 constituents were identified unambiguously or tentatively. Subsequently, the whole extract of the formula was separated into multiple components and the activity index method was used to calculate index values of the 79 constituents by integrating the chemical and pharmacological information of multiple components. Four polymethoxyl flavones were predicted as the major active constituents according to the activity index values. Furthermore, three polymethoxyl flavones were prepared using the strategy with semi-preparative LC guided by LC-MS, and their anti-inflammatory activities were validated. The results show that three polymethoxyl flavones with higher positive index values, i.e., 3, 5, 6, 7, 8, 3', 4'-heptamethoxyflavone, 3-hydroxynobiletein and tangeretin had significant anti-inflammatory effects. In conclusion, the predicted results indicated that the activity index method is feasible for the accurate prediction of active constituents in TCM formulae.

Analysis on the specificity of the relevant points along the governor vessel in chronic gastritis based on tenderness reaction and tenderness threshold / 中国针灸

OBJECTIVE@#To explore the specificity of the relevant points along the governor vessel in chronic gastritis through the synchronous observation of the tenderness reaction and tenderness threshold on the back along the governor vessel in the volunteers with chronic gastritis and make the comparison with healthy volunteers.@*METHODS@#A chronic gastritis group and a healthy volunteer group were prepared, 30 cases in each one. Using synchronous comparison, the tenderness reaction at the point inferior to each of the spinous processes from T to L along the governor vessel was observed. The mechanical pressure pain threshold was determined.@*RESULTS@#Regarding the tenderness reaction at the point inferior to each spinous process on the back along the governor vessel, the occurrence rate was different significantly between the chronic gastritis group and the healthy volunteer group (0.05).@*CONCLUSION@#The point inferior to the spoinous process of T on the governor vessel, Shendao (GV 11, T) and Lingtai (GV 10, T) are the points with the high specificity in chronic gastritis, which provides the reference to clinical treatment with acupuncture and moxibustion. The occurrence rate of high tenderness reaction and the low tenderness threshold may be the important manifestation of the point specificity on the back along the governor vessel in chronic gastritis.

Triterpenoids from the roots of Rubus parvifolius / 中国天然药物

Two new oleanane-type triterpenoids, parvifolactone A (1) and rubuside P (2), together with 11 known triterpenoids, fupenzic acid (3), 18,19-seco,2α,3α-dihydroxyl-19-oxo-urs-11,13(18)-dien-28-oic acid (4), euscaphic acid (5), maslinic acid (6), 1β- hydroxyeuscaphic acid (7), 2α,3α,19α,23-tetrahydroxyolean-12-en-28-oic acid (8), 2α,3β,19α,23-tetrahydroxyurs-12-en-28-oic acid (9), glucosyl pinfaensate (10), rubuside J (11), 2α,3α,19α,23-tetrahydroxyurs-12-en-24,28-dioic acid (12), and 2α,3β,19α- trihydroxyurs-12-en-23,28-dioic acid (13), were isolated from the roots of Rubus parvifolius.

Nuciferine, extracted from Nelumbo nucifera Gaertn, inhibits tumor-promoting effect of nicotine involving Wnt/ß-catenin signaling in non-small cell lung cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Nelumbo nucifera Gaertn are recorded in the earliest written documentation of traditional Chinese medicinal as "Ben Cao Gang Mu", a medicinal herb for blood clotting, dysentery and dizziness. Recently, nuciferine (NF), one of N. nucifera Gaertn leaf extracts has been shown to possess several pharmacological properties, including anti-viral and anti-cancer. The aim of this study is to investigate the underlying molecular mechanism of the anti-cancer activity of NF in NSCLC progression induced by nicotine MATERIALS AND METHODS: The effect of NF on proliferation of A549 (human lung adenocarcinoma epithelial cell line) pretreated with or without nicotine was detected by tumor cell proliferation assay. TOP-Flash reporter assay was applied to investigate the activity of Wnt/ß-catenin signaling in tumor cells in the presence of NF and/or nicotine. Apoptosis was measured using a FITC-Annexin V and PI detection kit by flow cytometry. In addition, mRNA or protein expression levels were respectively tested by quantitative RT-PCR or western blot. In vivo experiments, tumor samples were fixed in formalin and embedded in paraffin for additional analyses by immunohistochemistry and TUNEL staining. RESULTS: NF significantly inhibited the proliferation of NSCLC cells in the presence of nicotine, suppressed the activity of Wnt/ß-catenin signaling, enhanced the stabilization of Axin, and induced apoptosis. NF down-regulated the expression levels of ß-catenin and its downstream targets including c-myc, cyclin D and VEGF-A. NF also decreased the ratio of Bcl-2/Bax, which may explain the pro-apoptosis effect of NF. In tumor xenograft nude mice, NF not only inhibited the growth of non-small cell lung cancer (NSCLC) cells, but also remarkably alleviated the injury induced by nicotine in liver function. CONCLUSIONS: NF has the remarkable effect to inhibit nicotine-induced NSCLC progression, which was due to its ability to reduce the activity of Wnt/ß-catenin signaling. Thus, the work stated here emphasizes the importance of this traditional medicine and presents a potential novel alternative to NSCLC prevention and therapy.

Dimethyl Cardamonin Exhibits Anti-inflammatory Effects via Interfering with the PI3K-PDK1-PKCalpha Signaling Pathway

Consumption of herbal tea [flower buds of Cleistocalyx operculatus (Roxb.) Merr. et Perry (Myrtaceae)] is associated with health beneficial effects against multiple diseases including diabetes, asthma, and inflammatory bowel disease. Emerging evidences have reported that High mobility group box 1 (HMGB1) is considered as a key "late" proinflammatory factor by its unique secretion pattern in aforementioned diseases. Dimethyl cardamonin (2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone, DMC) is a major ingredient of C. operculatus flower buds. In this study, the anti-inflammatory effects of DMC and its underlying molecular mechanisms were investigated on lipopolysaccharide (LPS)-induced macrophages. DMC notably suppressed the mRNA expressions of TNF-alpha, IL-1beta, IL-6, and HMGB1, and also markedly decreased their productions in a time- and dose-dependent manner. Intriguingly, DMC could notably reduce LPS-stimulated HMGB1 secretion and its nucleo-cytoplasmic translocation. Furthermore, DMC dose-dependently inhibited the activation of phosphatidylinositol 3-kinase (PI3K), phosphoinositide-dependent kinase 1 (PDK1), and protein kinase C alpha (PKCalpha). All these data demonstrated that DMC had anti-inflammatory effects through reducing both early (TNF-alpha, IL-1beta, and IL-6) and late (HMGB1) cytokines expressions via interfering with the PI3K-PDK1-PKCalpha signaling pathway.

Identification of constituents in Suanzaoren tang by LC-Q-TOF-MS and LC-IT-MS / 中国中药杂志

LC-Q-TOF-MS and LC-IT-MS in positive and negative ion mode were applied to simultaneously characterize the constituents in Suanzaoren tang. Analysis was performed on an Agilent Zorbax SB-C18, Rapid Resolution HT column(4.6 mmx 50 mm, 1. 8 micro m) with gradient elution of acetonitrile(A) -aqueous solution containing 0. 05% formic acid(B) at a flow rate of 0. 6 mL min(-1) and the column temperature was 30 degreesC. By comparing MS fragmentation, accurate molecular weight, literature date and standard compounds information, a total of48 compounds were successfully identified or speculated. The origins of these compounds were assigned to the corresponding Chinese medicine. Thirty-one compounds were reported in Suanzaoren tang for the first time. LC-Q-TOF-MS combined with LC-IT-MS is a simple and rapid tool for the identification of constituents of Suanzaoren tang, and the results could provide evidence for the research on quality combined and effective constituents of Suanzaoren tang.

Chemical constituents from a Tibetan medicine Meconopsis horridula / 中国中药杂志

A phytochemical investigation on the aerial parts of a Tibetan medicine Meconopsis horridula, by solvent extraction, repeated chromatographies on silica gel, Sephadex LH-20, and preparative TLC techniques, led to the isolation of 9 compounds. By spectroscopic analysis and comparison of its 1H and 13C-NMR data with those in literatures, their structures were identified as oleracein E(1), N-( trans-p-coumaroyl) tyramine (2), chrysoeriol (3), apigenin (4), hydnocarpin (5), p-coumaric acid glucosyl ester (6), stigmast-5-ene-3beta-ylformate (7), 3beta-hydroxy-7alpha-ethoxy-24beta-ethylcholest-5-ene (8), and beta-sitosterol (9), respectively, among which compounds 6-8 were isolated from the genus for the first time,and 1,3 were isolated from the species for the first time. A MTT method was applied to evaluate the cytotoxic activity of compounds 14 against the human hepatocellular liver carcinoma cell line (HepG2), and compound 1 showed significant cytotoxicity against HepG2,with its inhibitory rate of 52.2% at 10 micromol x L(-1).

Effects of controllable dynamic inhaled exposure of moxa smoke on LDL-r, ICAM-1 and morphology of heart tissue in rats / 中国针灸

<p><b>OBJECTIVE</b>To observe the change of lipid metabolism and vascular endothelium as well as morphology of heart tissue in rats who were long-time exposed to moxa smoke with different concentrations in order to provide reference for safety assessment of moxa smoke on cardiovascular system.</p><p><b>METHODS</b>One hundred and sixty-eighty Wistar rats were randomly divided into a control group, a low-concentration group, a median-concentration group and a high-concentration group, 42 rats in each one. The rats were exposed to moxa smoke with concentration of 0%, 10%, 40% and 70%, respectively, for 20 min per day. After continuous intervention for six months, enzyme-linked immunosorbent assay (ELISA) was applied to measure the level of low density lipoprotein-receptor (LDL-r) and intercellular adhesion molecule-1 (ICAM-1) in blood serum in each group; the slices of heart tissue were stained with hematoxylin-eosin staining method to observe morphology change of heart tissue.</p><p><b>RESULTS</b>(1) After the intervention of moxa smoke, the levels of LDL-r and ICAM-1 in the low-concentration group were not statistically different from those in the control group (both P > 0.05); the level of LDL-r in the median-concentration group was significantly increased, which was statistically different from that in the control group [(3.87 +/- 0.27) mg/mL vs (2.12 +/- 0.13) mg/mL, P < 0.01], however, the content of ICAM-1 was not obviously changed; although the level of LDL-r in the high-concentration group was presented with an escalating trend, it was not statistically different from that in the control group (P > 0.05) while the level of ICAM-1 was obviously increased (P < 0.01). (2) Under the light microscope, the abnormalities of cardiac muscle fibers and myocardial cell in each group were not been observed.</p><p><b>CONCLUSION</b>The long-time intervention of low-concentration moxa smoke has no significant effects on lipid metabolism and vascular endothelium of rats, indicating that clinical application of low-concentration moxa smoke is relatively safe. The long-time intervention of moderate-concentration moxa smoke could significantly increase the clearance rate of cholesterol, implying the beneficial regulation of moxa smoke on lipid metabolism. The high-concentration moxa smoke could induce certain damage to vascular endothelium but its mechanism is in need of further research. The pathologic change of heart tissue could not be induced by moxa smoke with any concentration.</p>

Acupuncture Deqi Intensity and Propagated Sensation along Channels May, Respectively, Differ due to Different Body Positions of Subjects.

Acupuncture as an essential component of complementary and alternative medicine is gradually recognized and accepted by the mainstream of contemporary medicine. For obtaining preferable clinical effectiveness, Deqi is commonly regarded as efficacy predictor and parameter which is necessary to be achieved. Influential factors for acupuncture efficacy, like Deqi sensation as well as propagated sensation along channels (PSCs), enjoyed a long history in acupuncture basic research. Concerning this study, taking into account different positions on acupuncture Deqi sensation and PSCs, we would like to attest whether different body positions for subjects during needling procedure yield differed acupuncture Deqi sensation, particularly in terms of intensity, and PSCs. Methods. We used self-controlled method and selected 30 healthy subjects to perform needle insertion at Futu point (ST32) bilaterally. Then they were instructed to record the value of intensity of acupuncture sensation and the length and width of PSCs after removing the needle. Results. In regard to intensity of Deqi, kneeling seat position is stronger than supine position, accounting for 90% of the total number of subjects. In length of PSCs, kneeling seat position is greater than supine position, accounting for 56.7%. In width of PSCs, kneeling seat position is greater than supine position, accounting for 66.7%. Conclusion. Our findings show that needle inserting at Futu point (ST32) in kneeling seat position achieve better needle sensation and provide reference for clinical.

Effects of long-term intervention of moxa smoke on T lymphocyte subsets and CD4+ CD25+ Treg in peripheral blood of Wistar rats / 中国针灸

<p><b>OBJECTIVE</b>To investigate the cellular immune regulation of the long-term intervention of moxa smoke.</p><p><b>METHODS</b>Thirty-two Wistar rats were randomly divided into a blank group, a low concentration group, a medium concentration group and a high concentration group, 8 cases in each group. In addition to the blank group, rats in the other groups were exposed to the corresponding concentration moxa smoke for 20 min every day, the T lymphocyte subsets and proportion of the CD4+ CD25+ Treg in CD4+ T cells in peripheral blood were tested by flow cytometry after 6 months.</p><p><b>RESULTS</b>Compared with the blank group, the proportions of CD3+ CD4+, CD3+ CD8+ T cells and CD3+ CD4/CD3+ CD8+ in the other 3 moxa smoke groups were not significantly different (P > 0.05), while the proportions of the CD4+ CD25+ Treg in CD4+ T cells were significantly lower (P < 0.05), but no statistically significant differences among those 3 moxa smoke intervention groups (P > 0.05).</p><p><b>CONCLUSION</b>Long-term moxa smoke intervention has no significant effect on the proportions of CD3+ CD4+, CD3+ CD8+ T cells and CD3+ CD4+/CD3+ CD8+, but it can decrease the proportions of the CD4+ CD25+ Treg in CD4+ T cells in peripheral blood of rats. The way produced by pretreatment with moxa smoke may play immunomodulatory effect.</p>

Therapeutic effects of endoscopic therapy combined with enteral nutrition on acute severe biliary pancreatitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Acute severe biliary pancreatitis (ASBP) is a severe and fatal disease, and the expenditure is huge and therapeutic effects are still not satisfactory. This study aimed to improve the therapeutic effects and reduce the expenditure of ASBP treatment.</p><p><b>METHODS</b>One hundred and five patients diagnosed with ASBP were referred to our department from January 2004 to July 2009. Diagnosis was based on the 2007 criteria of the Chinese Society of Surgery. Patients were divided into two groups; the E group: 50 patients who underwent endoscopic retrograde choledochopancreatography (ERCP) + endoscopic sphincterotomy (EST) + endoscopic lithotripsy basket (ESR) + endoscopic retrograde biliary drainage (ERBD) and enteral nutrition (EN), and the R group: 55 patients who underwent traditional treatment without ERCP. Subsequently, subjective symptoms, signs, biochemical analysis, serum endotoxin, tumor necrosis factor a, grades by computed tomography (CT), cost of hospitalization and length of stay were compared between the two groups.</p><p><b>RESULTS</b>All enrolled patients complied well with all therapeutic regimens. Endoscopic therapy that combined EN could significantly improve symptoms, clinical signs, laboratory values, tumor necrosis factor a and endotoxin while significantly reducing hospital expenditure and length of hospital stay. The experimental findings revealed that there were obvious advantages in the E group compared with the R group.</p><p><b>CONCLUSIONS</b>Endoscopic therapy combined with EN is an effective, safe and economic therapeutic regimen of ASBP.</p>

Observation on therapeutic effects of elongated needle therapy on dysuria induced by benign prostatic hyperplasia / 中国针灸

<p><b>OBJECTIVE</b>To compare therapeutic effects of elongated needle therapy and routine acupuncture therapy on dysuria induced by benign prostatic hyperplasia (BPH).</p><p><b>METHODS</b>Randomized, controlled, multi-central method was adopted and 150 cases confirmed to the enrolled criteria were divided into two groups by odd or even number, an elongated needle group (n = 72) and a routine acupuncture group (n = 78). Acupuncture was given at bilateral Zhibian (BL 54) and Zhongji (CV 3) in the two groups, once daily, 5 sessions constituting one course, with a 2-day interval between two courses. The treatment was given for 2 courses. Changes of I-PSS symptom cumulative score, urine flowing rate, residual urine in bladder before and after the treatment were observed.</p><p><b>RESULTS</b>The effective rate was 83.3% in the elongated needle group and 44.9% in the routine acupuncture group. There were significant differences between the two groups in improvemet of I-PSS score, increase of urine flowing rate and reduction of residual urine in bladder (all P < 0.05).</p><p><b>CONCLUSION</b>The elongated needle therapy has a definite therapeutic effect on dysuria induced by benign prostatic hyperplasia.</p>

Role of T-cell receptor V beta 8.3 peptide vaccine in the prevention of experimental autoimmune uveoretinitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>T-cell receptor (TCR) plays an important role in the development of autoimmune diseases. Recently, it was reported that immunization of animals with TCR peptide derived from the pathogenic cells could prevent autoimmune diseases. The aim of this study was to investigate whether vaccination with a synthetic peptide from the hypervariable region of TCR V(beta) 8.3, an experimental autoimmune uveoretinitis (EAU)-associated gene, was able to prevent the disease.</p><p><b>METHODS</b>EAU was induced in Lewis rats by immunization with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA). The clinical and histological appearances were scored. Delayed type hypersensitivity (DTH) and lymphocyte proliferation were detected. Cytokine levels of aqueous humour, supernatants of cells from spleen and draining lymph nodes were measured by enzyme linked immunosorbent assay (ELISA). Gene expression of TCR V(beta) 8.3 on CD(4)(+) T cells was examined by real time quantitative polymerase chain reaction (PCR).</p><p><b>RESULTS</b>After vaccination, the intraocular inflammation was significantly mitigated, antigen specific DTH and lymphocyte proliferation responses were suppressed, interleukin (IL)-2 in aqueous humour, interferon (IFN)-gamma and IL-2 produced by the spleen and draining lymph node cells were significantly decreased, whereas the production of IL-4 and IL-10 were increased. The response of draining lymph node cells to TCR V(beta) 8.3 peptide was enhanced after vaccination. Inoculation with CFA alone did not affect the severity of EAU and the above parameters. The suppression of EAU was much stronger in the group of four fold inoculations than the group of two fold inoculations. The expression of TCR V(beta) 8.3 gene was significantly reduced in the group of fourfold inoculations.</p><p><b>CONCLUSION</b>Vaccination with the synthetic TCR V(beta) 8.3 peptide could remarkably inhibit the development of EAU.</p>