Amphotericin B for injection triggers degranulation of human LAD2 mast cells by MRGPRX2 and pseudo-allergic reactions in mice via MRGPRB2 activation.

Amphotericin B, a polyene macrolide antifungal agent, still plays an important role in the management of serious systemic fungal infections. Amphotericin B deoxycholate (AmBd) has been used to treat invasive fungal infections for over 60 years and remains the primary clinical formulation currently available. Anaphylactoid reactions triggered by AmBd in the clinic have been documented. However, the molecular and cellular events contributing to these reactions have not been clearly elucidated to date. This study demonstrates that the human Mas-related G protein-coupled receptor X2 (MRGPRX2) is the receptor that mediates these anaphylactoid responses. Molecular docking and cellular thermal shift assay (CETSA) indicate that AmBd exhibits potential affinity with MRGPRX2. In vitro, exposure to AmBd results in significant release of LAD2 mast cell granules and induces intracellular Ca2+ mobilization as well as activation of PLC-γ/IP3R and PI3K/AKT signaling pathways. However, these phenomena are reduced in MRGPRX2-knockdown LAD2 cells. In vivo, AmBd triggers paw swelling and a rapid drop in core body temperature in wild-type (WT) mice. However, these reactions are almost absent in MRGPRB2 (the mouse homolog of MRGPRX2) knockout mice (MRGPRB2MUT, MUT). The above results suggest that AmBd activates PLC-γ/IP3R and PI3K/AKT signaling via MRGPRX2 (in human LAD2 mast cells) or MRGPRB2 (in mice), leading to the release of mast cell granules and subsequent triggering of pseudo-allergic reactions. Taken together, this study clarifies the role of MRGPRX2 in triggering pseudo-allergic reactions to AmBd and suggests that MRGPRX2 could be a potential therapeutic target for controlling these reactions.

N-demethylsinomenine metabolite and its prototype sinomenine activate mast cells via MRGPRX2 and aggravate anaphylaxis.

Sinomenine hydrochloride (SH) is commonly used in the treatment of rheumatoid arthritis. It activates mast cells and induces anaphylaxis in the clinical setting. Adverse drug reactions can be caused by activation of MAS-associated G protein-coupled receptor X2 (MRGPRX2) on mast cells. Because the ligand binding site of MRGPRX2 is easily contacted in dilute solvents, it can be activated by many opioid drug structures. N-Demethylsinomenine (M-3) has a similar chemical structure to that of the opioid scaffold and is a major metabolite of SH. We sought to clarify whether M-3 induces anaphylaxis synergistically with its prototype in a mouse model. Molecular docking computer simulations suggested a similar binding effect between M-3 and SH. M-3 was chemically synthesized and analyzed by surface plasmon resonance to reveal its affinity for MRGPRX2. Temperature monitoring, in vivo hindlimb swelling and exudation test, and in vitro mast cell degranulation test were used to explore the mechanism of MRGPrx2 mediated allergic reaction triggered by M-3. Reduced M-3-induced inflammation was evident in MrgprB2 (the ortholog of MRGPRX2) conditional (Cpa3-Cre/MrgprB2flox) knockout (MrgprB2-CKO) mice. Additionally, LAD2 human mast cells with MRGPRX2 knockdown showed reduced degranulation. M-3 activated LAD2 cells synergistically with SH as regulated by GRK2 signaling and IP3R/PLC/PKC/P38 molecular signaling pathways. The results indicate that the M-3 metabolite can activate mast cells synergistically with its prototype SH via MRGPRX2 and aggravate anaphylaxis. These findings provide important insights into drug safety.

Acupuncture for chronic sciatica: protocol for a multicenter randomised controlled trial.

BACKGROUND: Chronic Sciatica is a disabling condition causing considerable medical, social and financial implications. Currently, there is no recognised long-term effective treatment to alleviate sciatica. Acupuncture has been widely used for treating chronic pains with persistent analgesic effects. We aim to evaluate the efficacy and safety of acupuncture for chronic sciatica with follow-up in 52 weeks. METHODS AND ANALYSIS: This is a multicenter randomised sham-controlled trial. A total of 216 patients with chronic sciatica will be enrolled and randomly assigned to the acupuncture or sham acupuncture group. There will be 10 treatment sessions applied in 4 weeks with frequency decreased over time. Patients will complete follow-ups during 52 weeks. The primary outcomes are changes in leg pain intensity and disability from baseline to week 4. Secondary outcomes include back pain intensity, frequency and bothersomeness, quality of life, and global perceived effect. Adverse events will be recorded in detail. ETHICS AND DISSEMINATION: Ethical approval of this trial was granted from the ethics committee of Beijing University of Chinese Medicine and all study centres (No. 2020BZYLL0803). Written informed consent will be obtained from enrolled patients. Trial results will be disseminated in peer-reviewed publications. TRIAL REGISTRATION NUMBER: ChiCTR2100044585 (Chinese Clinical Trial Registry, http://www.chictr.org.cn, registered on 24 March 2021); preresults.

Abnormal thymic stromal lymphopoietin expression in the gastrointestinal mucosa of patients with eosinophilic gastroenteritis / Expressão anormal da linfopoietina estromal tímica na mucosa gastrointestinal depacientes com gastroenterite e osinofílica

Abstract Objective: To investigate the differential expression of the thymic stromal lymphopoietin isoforms, short and long, and discern their biological implications under eosinophilic gastroenteritis. Methods: The expression of thymic stromal lymphopoietin and its two isoforms in tissues was assessed by quantitative RT-PCR in healthy controls (n = 24) and patients with eosinophilic gastroenteritis (n = 17). Results: Thymic stromal lymphopoietin mRNA was significantly reduced in eosinophilic gastroenteritis when compared with healthy controls (p < 0.0001). A significantly lower amount of short thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p < 0.05), while a significantly higher amount of long thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p < 0.05). Peak eosinophilic count is significantly positively correlated with the expression of long thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (r s = 0.623, p < 0.005), while peak eosinophilic count is significantly negatively correlated with the expression of short thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (r s = −0.4474, p < 0.05). Conclusions: Abnormal mucosal thymic stromal lymphopoietin expression may contribute to gastrointestinal mucosa damage in eosinophilic gastroenteritis.

Resumo Objetivo: Investigar a expressão diferencial das isoformas da linfopoietina estromal tímica, curta e longa, e discernir suas implicações biológicas na gastroenterite eosinofílica. Métodos: Avaliamos a expressão das isoformas da linfopoietina estromal tímica e suas duas isoformas através da técnica RT-PCR quantitativa em tecidos de controles saudáveis (n = 24) e pacientes com gastroenterite eosinofílica (n = 17). Resultados: Demonstramos que o RNAm das isoformas da linfopoietina estromal tímica estava significativamente reduzido na gastroenterite eosinofílica em comparação com os controles saudáveis (p < 0,0001). Também descobrimos uma quantidade significativamente menor de RNAm das isoformas da linfopoietina estromal tímica curta na gastroenterite eosinofílica em comparação com os controles (p < 0,05) e uma quantidade significativamente maior de RNAm das isoformas da linfopoietina estromal tímica longa na gastroenterite eosinofílica em comparação com os controles (p < 0,05). O pico da contagem eosinofílica está correlacionado positiva e significativamente com a expressão do RNAm das isoformas da linfopoietina estromal tímica longa na mucosa gastrointestinal de pacientes com gastroenterite eosinofílica (rs = 0,623, p < 0,005), enquanto o pico de contagem eosinofílica está negativa e significativamente correlacionado com a expressão do RNAm das isoformas da linfopoietina estromal tímica curta na mucosa gastrointestinal de pacientes com gastroenterite eosinofílica (rs = -0,4474, p < 0,05). Conclusões: A expressão anormal das isoformas da linfopoietina estromal tímica na mucosa pode contribuir para o dano da mucosa gastrointestinal na gastroenterite eosinofílica.

Abnormal thymic stromal lymphopoietin expression in the gastrointestinal mucosa of patients with eosinophilic gastroenteritis.

OBJECTIVE: To investigate the differential expression of the thymic stromal lymphopoietin isoforms, short and long, and discern their biological implications under eosinophilic gastroenteritis. METHODS: The expression of thymic stromal lymphopoietin and its two isoforms in tissues was assessed by quantitative RT-PCR in healthy controls (n=24) and patients with eosinophilic gastroenteritis (n=17). RESULTS: Thymic stromal lymphopoietin mRNA was significantly reduced in eosinophilic gastroenteritis when compared with healthy controls (p<0.0001). A significantly lower amount of short thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p<0.05), while a significantly higher amount of long thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p<0.05). Peak eosinophilic count is significantly positively correlated with the expression of long thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (rs=0.623, p<0.005), while peak eosinophilic count is significantly negatively correlated with the expression of short thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (rs=-0.4474, p<0.05). CONCLUSIONS: Abnormal mucosal thymic stromal lymphopoietin expression may contribute to gastrointestinal mucosa damage in eosinophilic gastroenteritis.

Acupuncture or metformin to improve insulin resistance in women with polycystic ovary syndrome: study protocol of a combined multinational cross sectional case-control study and a randomised controlled trial.

INTRODUCTION: Polycystic ovary syndrome (PCOS) is linked to hyperinsulinemia and insulin resistance with dysfunctional glucose metabolism. Pilot studies suggests that acupuncture treatment with combined manual and low-frequency electrical stimulation (electroacupuncture (EA)) of the needles decrease circulating glycated haemoglobulin (HbA1c) and homeostatic model assessment-insulin resistance. Therefore, we here aim to investigate if acupuncture treatment or metformin together with lifestyle or lifestyle management alone improves insulin sensitivity and related symptoms in overweight/obese women with PCOS. METHODS AND ANALYSIS: This is a two-centre multinational (Sweden and China), cross-sectional case-control study combined with an open-labelled randomised controlled trial (RCT). Participants are randomised to one of three groups: (1) EA 2-3 times/week during 4 months+lifestyle management; (2) metformin, 500 mg, three/day during 4 months+lifestyle management; or (3) lifestyle management alone. The primary outcome measure in the RCT is changes in HbA1C. A total of 123 obese overweight women with PCOS will be enrolled and randomised into one of the three groups with a target power of at least 80% and 5% significance level based on two-sided tests. ETHICS AND DISSEMINATION: The study has been approved by the Regional Ethical Review Board of Stockholm and of Peking University Third Hospital, China. Primary outcome data of the RCT will be published in a relevant journal together with supporting secondary outcome measurements. Further, outcome measurements will be published in separate papers as well as case-control data. EXPECTED RESULTS: We anticipate that EA and metformin, both with lifestyle management, are equally effective and superior to lifestyle management alone for improvement of glycaemic control. TRIAL REGISTRATION NUMBERS: NCT02647827 and EudraCT2015-004250-18.

Effects of Chinese herbs combined with in vitro fertilization and embryo transplantation on infertility: a clinical randomized controlled trial.

OBJECTIVE: To assess the effects of using Chinese herbs in assisted reproductive technology. METHODS: Four hundred and thirty-three subjects aged less than 42 years with infertility due to Fallopian tube or male-related factors who were willing to undertake in vitro fertilization and embryo transplantation were randomly allocated to a Chinese herb intervention group (n = 216) or a conventional treatment control group (n = 217). All subjects received one of four routine ultra-ovulation-promoting therapies at the Reproductive Center in the Third Hospital Affiliated to Beijing University according to their physician's assessments. The subjects in the intervention group received various Chinese herbs depending on their conventional treatment. Endometrial thickness, number of acquired eggs, and rates of normal fertility, high-quality embryos, biochemical and clnical pregnancy of subjects were assessed in both groups. RESULTS: The high-quality embryo rate of 51.9%, biochemical pregnancy rate of 51.0%, clinical pregnancy rate of 44.2% and endometrial thickness of (10.84 +/- 1.75) mm in the intervention group were all significantly higher than those in the control group [48.7%, 38.9%, 34.8%, and (10.52 +/- 1.50) mm, respectively; P < 0.05]. The normal fertility rate of 58.5% in the Chinese herb group was also significantly superior to the 54.7% achieved in the control group (P < 0.01). There were no statistically significant differences (P > 0.05) in the average number of acquired eggs within a single cycle, incidence of excessive stimulation of ovary, rates of embryo transplantation or early abortion and birth of living babies between the two groups. CONCLUSION: Our findings indicate that Chinese herbs increase endometrial thickness, improve the quality of fertility and embryo, and promote embryonic nidation, thus enhancing the success rate of in vitro fertilization/intracytoplasmic sperm injection-embryo transplantation cycle. Using Chinese herbs improves the outcomes and safety of assisted reproductive technologies.

[Protective effect of acupuncture on heart in mice with hyperlipemia and its mechanism].

OBJECTIVE: To observe the inhibiting effect of acupuncture on blood lipid, myocardial hypertrophy and fibrosis in mice with hyperlipemia, and explore its possible action mechanism. METHODS: Ten inbred mice (C57) were applied. Forty ApoE(-/-) mice who removed gene of apolipoprotein E were randomly divided into a control group, a non-acupoint group, an acupoint group and a medication group. The points 0. 5 cm and 1 cm next to the end of mice tail were respectively punctured in the non-acupoint group; "Neiguan" (PC 6) and "Fenglong" (ST 40) were punctured in the acupoint group; intragastric administration of simvastatin was applied in the medication group. After 8 weeks of treatment, the changes of total cholesterol (TC) and ratio of heart to body mass in each group were measured; changes of cardiac muscle fiber and ventricular wall thickness were observed; enzyme linked immunosorbent assay (ELISA) was used to test the level of angiotensin II (Ang I ) in plasma, and western blotting method was used to test protein content of angiotensin II type 1 receptor (AT1R) and endothelin-1 type A receptor (ETAR) in the heart. RESULTS: After 8 weeks of intervention, compared with the control group, rising range of blood lipid was obviously decreased (P<0.01) in the acupoint group and medication group, ratio of P<0.01), myocardial heart to body mass was decreased (P<0.05), thickness of ventricular wall was reduced (P fibrosis was relieved, levels of Ang II and ET-1 in plasma were decreased (P<0. 05), content of NO was increased (P<0. 05), and protein content of AT1R and ETAR was decreased in the heart (P<0. 05). CONCLUSION: 40) could inhibit the rising of blood lipid in ApoE(-/-) mice, lower the levels of Ang II and ET-1 in peripheral blood, increase the content of NO and inhibit the expression of AT1R and ETAR in heart tissue, which could relieve myocardial hypertrophy and fibrosis to play a protective role on heart.

Determination of sRNA expressions by RNA-seq in Yersinia pestis grown in vitro and during infection.

BACKGROUND: Small non-coding RNAs (sRNAs) facilitate host-microbe interactions. They have a central function in the post-transcriptional regulation during pathogenic lifestyles. Hfq, an RNA-binding protein that many sRNAs act in conjunction with, is required for Y. pestis pathogenesis. However, information on how Yersinia pestis modulates the expression of sRNAs during infection is largely unknown. METHODOLOGY AND PRINCIPAL FINDINGS: We used RNA-seq technology to identify the sRNA candidates expressed from Y. pestis grown in vitro and in the infected lungs of mice. A total of 104 sRNAs were found, including 26 previously annotated sRNAs, by searching against the Rfam database with 78 novel sRNA candidates. Approximately 89% (93/104) of these sRNAs from Y. pestis are shared with its ancestor Y. pseudotuberculosis. Ninety-seven percent of these sRNAs (101/104) are shared among more than 80 sequenced genomes of 135 Y. pestis strains. These 78 novel sRNAs include 62 intergenic and 16 antisense sRNAs. Fourteen sRNAs were selected for verification by independent Northern blot analysis. Results showed that nine selected sRNA transcripts were Hfq-dependent. Interestingly, three novel sRNAs were identified as new members of the transcription factor CRP regulon. Semi-quantitative analysis revealed that Y. pestis from the infected lungs induced the expressions of six sRNAs including RyhB1, RyhB2, CyaR/RyeE, 6S RNA, RybB and sR039 and repressed the expressions of four sRNAs, including CsrB, CsrC, 4.5S RNA and sR027. CONCLUSIONS AND SIGNIFICANCE: This study is the first attempt to subject RNA from Y. pestis-infected samples to direct high-throughput sequencing. Many novel sRNAs were identified and the expression patterns of relevant sRNAs in Y. pestis during in vitro growth and in vivo infection were revealed. The annotated sRNAs accounted for the most abundant sRNAs either expressed in bacteria grown in vitro or differentially expressed in the infected lungs. These findings suggested these sRNAs may have important functions in Y. pestis physiology or pathogenesis.

[Intervention of controlled ovarian hyperstimulation cycle by Chinese medical herbs for Shen tonifying, blood nourishing and activating: a randomized clinical trial].

OBJECTIVE: To study the effect of Chinese medical herbs for Shen tonifying, blood nourishing and activating (CMHSTBNA) on the cycle of controlled ovarian hyperstimulation (COH) of assisted reproductive technique (ART). METHODS: A large sample randomized control trial was performed. Infertility women patients, younger than 42 years (infertility due to tubal factor and/or male factor) were randomly assigned to the CMHSTBNA intervention group (abbreviated as the treated group) and the control group, 184 cases in each group. All underwent COH. Those in the treated group received assist therapy of CMHSTBNA from the menstrual period day 2 -3 of COH to the day of oocytes retrieved. The serum hormone level [including estrogen (E2), progesterone(P), luteal hormone (LH) on the day of human chorionic gonadotrophin (hCG) administration], the medication days and dosage of gonadotropin (Gn), the number of oocytes retrieved, the fertilization rate, and the good-quality embryo rate were observed and compared with the control group. RESULTS: The endometrial thickness on the day of oocytes retrieved was 10.85 +/- 1.63 mm in the treated group, larger than that in the control group (10.50 +/- 1.49 mm) (P <0.05). The good-quality embryo rate and the frozen rate were 48. 9% and 39. 7% respectively in the treated group, superior to those of the control group (45. 4% and 35. 8% respectively), showing statistical difference (P < 0.05). On the day of hCG administration, favorable tendency was shown in the serum levels of estradial (E2), progesterone (P), luteinizing hormone (LH), the medication days and dosage of Gn, the number of oocytes retrieved, the fertilization rate, and the cleavage rate, showing no statistical difference when compared with the control group (P >0.05). CONCLUSION: The combined application of CMHSTBNA and gonado-trophic hormones in COH cycle could elevate the embryo quality, improve the endometrial state, thus laying foundation for successful in vitro fertilization/intracytoplasmic sperm embryo transfer.

Two sRNA RyhB homologs from Yersinia pestis biovar microtus expressed in vivo have differential Hfq-dependent stability.

Small non-coding RNAs (sRNAs) have been shown to modulate gene expression at the post-transcriptional level. RyhB, an iron-responsive sRNA, is conserved in Escherichia coli and other Enterobacteriae, indicating the downregulation of numerous genes during iron depletion. This sRNA is tightly regulated by the ferric uptake regulator (Fur) and interacts with the RNA binding protein Hfq. Hfq is generally purported to be essential for stabilizing sRNAs and promoting sRNA-mRNA duplex formation. Maintenance of iron homeostasis is an essential step in the lifecycle of Yersinia pestis. Y. pestis encodes two RyhB homologs, RyhB1 and RyhB2. In this study, we found that as in the case of E. coli, both RyhB homologs in Y. pestis are negatively regulated by Fur and have a half-life of >30 min. In the absence of Hfq, RyhB1 is rapidly degraded, while RyhB2 retains its stability. RyhB1 stabilization is mediated by Hfq, but RyhB2 does not require Hfq for stability. Additionally, both RyhBs are upregulated in lungs infected with Y. pestis, while the ryhB mutant shows no visible effects on virulence in mice upon either subcutaneous or intranasal inoculation. Collectively, our results indicate that the two RyhB homologs have common regulatory features in Y. pestis-infected lungs and in vitro, but that stability of RyhB1 and RyhB2 is differentially dependent on Hfq.

A ratiometric fluorescence sensor for Be2+ based on Beryllon II/layered double hydroxide ultrathin films.

A ratiometric fluorescence sensor for Be(2+) has been fabricated via alternate assembly of 2-(3,6-disulfo-8-hydroxynaphthylazo)-1,8-dihydroxynaphthalene-3,6-disulfonate (Beryllon II) and MgAl-LDH nanosheets on quartz substrates using the layer-by-layer (LBL) deposition technique. UV-vis absorption and the fluorescence emission spectroscopy indicate a stepwise and regular growth of the Beryllon II/LDH UTFs upon increasing deposition cycle. The film of Beryllon II/LDH possesses a periodic layered structure perpendicular to the substrate revealed by X-ray diffraction and scanning electron microscopy. Atomic force microscopy images show that the film surface is continuous and uniform. The Beryllon II/LDH UTFs display ratiometric fluorescence response for Be(2+) with a linear response range in 1.0×10(-7)-1.9×10(-6) mol L(-1) and a detection limit of 4.2×10(-9) mol L(-1). Furthermore, the ratiometric sensor exhibits good repeatability, high stability (thermal, storage and mechanical) as well as excellent selectivity toward Be(2+). XPS and Raman measurements demonstrate that the specific response of the sensor is attributed to the coordination between Be(2+) and Beryllon II in the UTF. The Beryllon II/LDH UTFs in this work can be potentially used as a chemosensor for the detection of Be(2+) in the environmental and biomedical field.

Resettable fluorescence logic gate based on calcein/layered double hydroxide ultrathin films.

A fluorescent logic gate was fabricated based on calcein/layered double hydroxide ultrathin films (UTFs) via alternate assembly technique, which exhibits high stability, reversibility, and resettability. The logic gate was manipulated by utilizing pH value, Hg(2+) and Cl(-) ion as inputs, and the fluorescence emission of the (calcein/LDH)(16) UTF as output, serving as a three-input logic gate that combines the YES and INHIBIT operation.

Identification by cDNA cloning of abundant sRNAs in a human-avirulent Yersinia pestis strain grown under five different growth conditions.

AIMS: sRNA regulation is supposedly involved in the stress response of a pathogen during infection. Yersinia pestis, the etiologic agent of plague, must encounter temperature and microenvironment changes, given its lifestyle. Here, we used the cDNA cloning approach to discover full-length sRNA candidates that are highly expressed in Y. pestis under five different growth conditions. MATERIALS & METHODS: The cDNA cloning approach was improved by combining the traditional cDNA library construction with the prevalent rapid amplification of cDNA ends and RNA size selection techniques. RESULTS: In total, 43 RNA species, including six previously annotated sRNAs, were identified. Of these, 25 sRNAs were encoded on the antisense strand of the annotated genes. Interestingly, two of these sRNAs were found on the complementary strand of noncoding RNAs. In addition, eight novel sRNAs encoded in the intergenic regions were also revealed. Ten sRNA candidates chosen for the northern blot analysis were successfully detected. Analysis of the expression patterns of 29 candidate sRNAs showed that 24 sRNAs are highly abundant in Y. pestis upon entry into the stationary growth phase. CONCLUSION: Our preliminary attempt at screening the novel sRNA candidates will lay the foundation for understanding the roles of sRNAs in Y. pestis physiology and pathogenesis.

OmpR may regulate the putative YehU/YehT two-component system in Salmonella enterica serovar Typhi under hypotonic growth condition.

Decreased expression (twofold) of a putative yehUTS operon of which yehUT encodes a putative YehU/YehT two-component system in the ompR mutant from Salmonella enterica serovar Typhi (S. Typhi) GIFU10007 under hypotonic growth condition was observed by qRT-PCR. Purified recombinant protein OmpR(His6) of GIFU10007 was shown to bind the upstream region of the yehU gene by the gel-shift assay. In addition, the yehT deletion mutant (ΔyehT) displayed differential expression (twofold or higher) of 26 genes under the condition by the DNA microarray analysis. Altogether, OmpR might regulate the YehUT system in S. Typhi under hypotonic growth condition.

[Study on vesicant characteristics of Uylabris and seeds of Brassica Alb].

UNLABELLED: OBJECTIVE To study on vesicant characteristics of Uylabris and Seeds of Brassica Alb in crude herb moxibustion. METHODS: One hundred and seventy-five healthy subjects were randomly divided into 5 groups. The vesicant rate, vesicant time and vesicant size and pain degree of different medicines or doses were observed. Results There were significant differences in the vesicant rate among the different doses groups of the same medicine (P<0.05), with no significant difference in the vesicant time (P>0.05); and there were significant differences in vesicant time among the different medicine groups (P<0.05). In the Uylabris group, the vesicant time was longer; the vesicant size in the medicine group of 0. 05 g each point was larger than that of the medicine group of 0.01 g each point (P<0.05); there were significant differences among the groups in the pain rate (P<0.05), and the pain rate was the highest in the 0.05 g each point of Seeds of Brassica Alb. CONCLUSION: Uylabris and Seeds of Brassica Alb have different vesicular characteristics, so in clinical application, application dose and time may be used for controlling the vesicant extent.