Deciphering chemical and metabolite profiling of Chang-Kang-Fang by UPLC-Q-TOF-MS/MS and its potential active components identification / 中国天然药物

Chang-Kang-Fang (CKF) formula, a Traditional Chinese Medicine (TCM) prescription, has been widely used for the treatment of irritable bowel syndrome (IBS). However, its potential material basis and underlying mechanism remain elusive. Therefore, this study employed an integrated approach that combined ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) with network pharmacology to systematically characterize the phytochemical components and metabolites of CKF, as well as elucidating its underlying mechanism. Through this comprehensive analysis, a total of 150 components were identified or tentatively characterized within the CKF formula. Notably, six N-acetyldopamine oligomers from CicadaePeriostracum and eight resin glycosides from Cuscutae Semen were characterized in this formula for the first time. Meanwhile, 149 xenobiotics (58 prototypes and 91 metabolites) were detected in plasma, urine, feces, brain, and intestinal contents, and the in vivo metabolic pathways of resin glycosides were elaborated for the first time. Furthermore, network pharmacology and molecular docking analyses revealed that alkaloids, flavonoids, chromones, monoterpenes, N-acetyldopamine dimers, p-hydroxycinnamic acid, and Cus-3/isomer might be responsible for the beneficial effects of CKF in treating IBS, and CASP8, MARK14, PIK3C, PIK3R1, TLR4, and TNF may be its potential targets. These discoveries offer a comprehensive understanding of the potential material basis and clarify the underlying mechanism of the CKF formula in treating IBS, facilitating the broader application of CKF in the field of medicine.

Activation-induced cytidine deaminase (AID) involved in the regulation of B cell immune senescence / 细胞与分子免疫学杂志

The humoral immune response of B cells is the key to the protection of specific immunity, and immune aging reshapes its production and function. The decreased B cell immune function is an indicator of immune senescence. The impaired humoral immune function mediated by antibody secreted by B cells leads to a decline in the response of elderly individuals to the vaccine. These people are therefore more susceptible to infection and deterioration, and have a higher incidence of tumors and metabolic diseases. Activation-induced cytidine deaminase (AID) is an enzyme that triggers immunoglobulin class conversion recombination (CSR) and somatic high frequency mutation (SHM). It decreases during immune senescence and is considered to be a biomarker of decreased B cell function in aging mice and humans. Understanding the inherent defects of B-cell immune senescence and the regulation mechanism of AID in the aging process can provide new research ideas for the susceptibility, prevention and treatment of diseases in the elderly.

Identifying the molecular basis of Jinhong tablets against chronic superficial gastritis via chemical profile identification and symptom-guided network pharmacology analysis / 药物分析学报

Chronic superficial gastritis(CSG)is a common disease of the digestive system that possesses a serious pathogenesis.Jinhong tablet(JHT),a traditional Chinese medicine(TCM)prescription,exerts therapeutic effects against CSG.However,the molecular basis of its therapeutic effect has not been clarified.Herein,we employed ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q/TOF-MS)based chemical profile identification to determine the chemical components in JHT.Further,we applied network pharmacology to illustrate its molecular mechanisms.A total of 96 chemical constituents were identified in JHT,31 of which were confirmed using reference standards.Based on the bioinformatics analysis using the symptom-guided pharmacological networks of"chi,""blood,""pain,"and"inflammation,"and target screening through the interaction probabilities between compounds and targets,matrix metalloproteinase 2(MMP2),dopamine d2 receptor(DRD2),and Aldo-keto reductase family 1 member B1(AKR1B1)were identified as key targets in the therapeutic effect exhibited by JHT against CSG.Moreover,according to the inhibitory activities presented in the literature and binding mode analysis,the structural types of alkaloids,flavonoids,organic acids,including chlorogenic acid(10),caffeic acid(13),(-)-corydalmine(33),(-)-isocorypalmine(36),isochlorogenic acid C(38),isochlorogenic acid A(41),quercetin-3-O-α-L-rhamnoside(42),isochlorogenic acid B(47),quercetin(63),and kaempferol(70)tended to show remarkable activities against CSG.Owing to the above findings,we systematically identified the chemical components of JHT and revealed its molecular mechanisms based on the symptoms associated with CSG.

Chemical screen identifies shikonin as a broad DNA damage response inhibitor that enhances chemotherapy through inhibiting ATM and ATR

DNA damage response (DDR) is a highly conserved genome surveillance mechanism that preserves cell viability in the presence of chemotherapeutic drugs. Hence, small molecules that inhibit DDR are expected to enhance the anti-cancer effect of chemotherapy. Through a recent chemical library screen, we identified shikonin as an inhibitor that strongly suppressed DDR activated by various chemotherapeutic drugs in cancer cell lines derived from different origins. Mechanistically, shikonin inhibited the activation of ataxia telangiectasia mutated (ATM), and to a lesser degree ATM and RAD3-related (ATR), two master upstream regulators of the DDR signal, through inducing degradation of ATM and ATR-interacting protein (ATRIP), an obligate associating protein of ATR, respectively. As a result of DDR inhibition, shikonin enhanced the anti-cancer effect of chemotherapeutic drugs in both cell cultures and in mouse models. While degradation of ATRIP is proteasome dependent, that of ATM depends on caspase- and lysosome-, but not proteasome. Overexpression of ATM significantly mitigated DDR inhibition and cell death induced by shikonin and chemotherapeutic drugs. These novel findings reveal shikonin as a pan DDR inhibitor and identify ATM as a primary factor in determining the chemo sensitizing effect of shikonin. Our data may facilitate the development of shikonin and its derivatives as potential chemotherapy sensitizers through inducing ATM degradation.

Extensive expansion of the chemical diversity of fusidane-type antibiotics using a stochastic combinational strategy

Fusidane-type antibiotics, represented by helvolic acid, fusidic acid and cephalosporin P

Research progress in characteristics of B cells in high-, low- and non-responsive individuals after HBV vaccination / 中华微生物学和免疫学杂志

Hepatitis B virus (HBV) infection remains a serious global health problem, which can lead to acute and chronic liver diseases and a variety of complications. HBV vaccination, which induces B cells to secrete protective hepatitis B surface antibody (HBsAb), is an important measure to prevent HBV infection. Some studies have shown that different individuals have different responses to HBV vaccine, which can be divided into ultra-high/high, normal/medium and low/non-response. Research on the potential mechanisms can provide reference for the preparation of high-titer HBsAb and the prevention and treatment of HBV infection. This paper reviewed the research situation and progress in the characteristics and mechanism of B cells in different responders after HBV vaccination.

Tripodalsporormielones A-C, unprecedented cage-like polyketides with complex polyvdent bridged and fused ring systems

A chemical investigation on

A four-protein metabolon assembled by a small peptide protein creates the pentacyclic carbonate ring of aldgamycins

Organic carbonates (OCs) are a class of compounds featured by a carbonyl flanked by two alkoxy/aryloxy groups. They exist in either linear or cyclic forms, of which the majority encountered in nature adopt a pentacyclic structure. However, the enzymatic basis for pentacyclic carbonate ring formation remains elusive. Here, we reported that a four-protein metabolon (AlmUII-UV) assembled by a small peptide protein (AlmUV) appends a reactive

Biosynthesis of clinically used antibiotic fusidic acid and identification of two short-chain dehydrogenase/reductases with converse stereoselectivity

Fusidic acid is the only fusidane-type antibiotic that has been clinically used. However, biosynthesis of this important molecule in fungi is poorly understood. We have recently elucidated the biosynthesis of fusidane-type antibiotic helvolic acid, which provides us with clues to identify a possible gene cluster for fusidic acid ( cluster). This gene cluster consists of eight genes, among which six are conserved in the helvolic acid gene cluster except and . Introduction of the two genes into the NSAR1 expressing the conserved six genes led to the production of fusidic acid. A stepwise introduction of and revealed that the two genes worked independently without a strict reaction order. Notably, we identified two short-chain dehydrogenase/reductase genes and in the cluster, which showed converse stereoselectivity in 3-ketoreduction. This is the first report on the biosynthesis and heterologous expression of fusidic acid.

Inhibition of Rac1-dependent forgetting alleviates memory deficits in animal models of Alzheimer's disease

Accelerated forgetting has been identified as a feature of Alzheimer's disease (AD), but the therapeutic efficacy of the manipulation of biological mechanisms of forgetting has not been assessed in AD animal models. Ras-related C3 botulinum toxin substrate 1 (Rac1), a small GTPase, has been shown to regulate active forgetting in Drosophila and mice. Here, we showed that Rac1 activity is aberrantly elevated in the hippocampal tissues of AD patients and AD animal models. Moreover, amyloid-beta 42 could induce Rac1 activation in cultured cells. The elevation of Rac1 activity not only accelerated 6-hour spatial memory decay in 3-month-old APP/PS1 mice, but also significantly contributed to severe memory loss in aged APP/PS1 mice. A similar age-dependent Rac1 activity-based memory loss was also observed in an AD fly model. Moreover, inhibition of Rac1 activity could ameliorate cognitive defects and synaptic plasticity in AD animal models. Finally, two novel compounds, identified through behavioral screening of a randomly selected pool of brain permeable small molecules for their positive effect in rescuing memory loss in both fly and mouse models, were found to be capable of inhibiting Rac1 activity. Thus, multiple lines of evidence corroborate in supporting the idea that inhibition of Rac1 activity is effective for treating AD-related memory loss.

Effectiveness and safety of interleukin-2 plus cisplatin for treating malignant pleural effusion:a meta analysis / 重庆医学

Objective To systematically evaluate the effectiveness and safety of interleukin-2 plus cisplatin for treating ma-lignant pleural effusion(MPE)to provide a basis for clinical treatment strategy.Methods CBM,CNKI,VIP,Wanfang,Pubmed, Embase,Cochrane library clinical trial registration database were systematically retrieved.The randomized controlled trial(RCT) quality assessment criteria of Cochrane collaboration network was adopted for including the study quality.The data were extracted by meta analysis.Results (1)Thirty-four RCT involving 2 037 MPE patients were included,the quality of included RCT was ordi-nary;(2)compared with simple cisplatin,the merged RR values and their 95%CI of meta-analysis for ORR,fever,were 1.45 (1.36-1.54),2.37 (1.53 -3.66),respectively,the differences between the two groups were statistically significant(P 0.05).Conclusion This study indicates that IL-2 plus cisplatin can significantly improve the clinical curative effect in the pa-tients with MPE,but has the adverse reactions of fever,etc.and the quality of included RCT is general.

Spectrum drift characteristics of CD4+ CD25+ Tregs TCR CDR3 in hepatitis B patients before and after treatment / 中国免疫学杂志

Objective:To determine the spectrum drift characteristics of CI4+CD25+Tregs TCR β chain CDR3 in patients with different phases of acute hepatitis B (AHB) and chronic hepatitis B (CHB) patients before and after the entecavir treatment.Methods:Anticoagulation venous blood was collected from 4 normal control subjects,3 AHB patients with acute phase and convalescent phase,and 4 CHB patients before and after the entecavir treatment;and peripheral blood mononuclear cells were isolated;CD4+ CD25+ Tregs were separated by using the magnetic beads,and total RNAs were extracted from CD4+ CD25+ Tregs and used for reverse transcription.The TRBV CDR3 was amplified by polymerase chain reaction (PCR) with forward primers specific for 24 TRBV families and one fluorescence-labeled common reverse primer specific for the BC region.The PCR products were sent out for Genescan,and results were analyzed for the TRBV family CDR3 spectrum characteristics by using the Peak Scanner Software vl.0.Data were analyzed with the comparative t-test to perform the statistical analysis.Results:The CDR3 spectral types of the TRBV family showed drift characteristics in 3 cases of AHB patients with acute and convalescent phases;single/oligo peak spectral type family was observed in most of patients with acute phase;multiple peak spectral type was seen in patients with convalescent phase;and the common spectrum shift of TRBV4,10,14,16,19 families seen in patients with acute phase was changed to multiple peak spectral type.The clonal expansion of TRBV family in the CD4+CD25+Tregs in PBMC from AHB patients with convalescent phase was significantly lower than AHB patients with acute phase (t =9.456,P =0.011).The clonal expansion of Tregs TRBV13.2,15,16,18,20 family seen in C HB patients before treatment may interfere the virus removal through down-regulating the body's immune response;and with the decline of viral load in serum after the antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24 family may help body induce immune tolerance and result in the HBV persistence.The clonal expansion of TRBV family in the CI4+CD25+Tregs in PBMC from of CHB patients after antiviral treatment was increased (t =-0.666,P =0.553).Conclusion:TRBV4,10,14,16,19 family of spectrum shift seen in AHB patients with acute phase was changed to multiple peak spectral type in patients with convalescent phase,suggesting this transition may be associated with HBsAg and HBeAg turning to negative.The clonal expansion of Tregs TRBV13.2,15,16,18,20 family seen in CHB patients before treatment may interfere the virus removal through down-regulating the body's immune response;and with the decline of viral load in serum after the antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24 family may help body induce immune tolerance and result in the HBV persistence.

A set of interesting sequoiatones stereoisomers from a wetland soil-derived fungus

Four interesting sequoiatones stereoisomers (-) were isolated from a wetland soil-derived fungusby chiral HPLC. On the basis of comprehensive NMR and mass analyses, their planar structures were elucidated as the same as that of sequoiatone B. Among them,and(orand) were a pair of enantiomers, andand(orand) were a pair of stereoisomers with epimerization at C-12, which indicated that sequoiatione-type metabolites exist as enantiomers rather than as optically pure compounds in some strains. With the quantum chemical ECD calculations, the absolute configurations of C-8 in-were determined, which is the first report to establish the absolute configuration of C-8 in sequoiatones. However, the absolute configurations of C-12 in sequoiatones are still unsolved.

Characterization of naive and memory T cell after immunization with H1N1 influenza vaccine for children / 中国免疫学杂志

Objective:To elucidate the characterization of CD8+T cell in H1N1 influenza vaccine for children.Methods:PBMCs were isolated from 31 children aged from 3 to 6 years old who had accepted H 1N1 influenza vaccine during December 2009 to January 2010.The lymphocytes were joined with the H 1N1 influenza vaccine as experimental group and cultured .The experiment set without vaccine group as control group .At last we detected the surface molecules by FCM .The CCK-8 assay was added to detecting cellular proliferation and cellular proliferation index were detected by CCK-8.Results: CD8+T cells of PBMC in the two groups were 13.41%and 9.41%,P>0.05.CD8+CD45RAA+naive T cells in the two groups were up to more than 80%,P>0.05.The proportion of CD8+CD45ROA+memory T cells in two groups were up to 17%-19%,P>0.05.Two subsets of CD8+CD45ROA+memory T cells :CCR7+and CD62L+single positive memory T cell subsets in the experimental group were significantly lower than that of the control group,P<0.05.The CCK-8 assay was added to detect cellular proliferation .Only 51.16% of which cellular proliferation index was greater than 0.8,with none was greater than 1 in this study.Conclusion:This study showed that the CD4+T cells were low-level,naive T cells (CD8+CD45RAA+)were higher,with antigen stimulation and response.H1N1 vaccination specific memory T cells were few in number , specific memory T cell subsets were diversity , control memory cells were the main phenotypic characteristics .Cellular proliferation index showed that the proliferation of specific CD 8+T cells vaccine was poor .

Analysis of BCR CDR3 repertoire of peripheral blood with HBsAb titer higher than 10 000 mU/ml / 中国免疫学杂志

Objective:To acquire potential HBsAb sequences,we have analyzed the BCR CDR3 repertoire of the peripheral blood with HBsAb titer higher than 10 000 mU/ml,which could provide a data basis for follow-up study.Methods:Genomic DNA of pe-ripheral blood mononuclear cells was extracted from samples with HBsAb titer higher than 10 000 mU/ml.We have adopted Illumina Solexa high-throughput sequencing technology of the Adaptive Biotechnologies ImmunoSEQ platform to acquire sequence data.IMGT/High V-QUEST was used to preliminary analyze our sequence data,including usage of IGHV,IGHJ and IGHD gene subgroups,IGHV-J matching,distribution of CDR3 amino acid (AA) length and usage of total CDR3 AA.And these sequences were compared with the HBsAb sequences from NCBI database.Results:Experimental samples have highly selected gene subgroups IGHV3,IGHV4,IGHJ4, IGHJ6,IGHD3,IGHD6,and IGHV3-J4 pairing,IGHV3-J6 pairing.The AA length distributions of CDR3 region were normal distribution with the length of 14/15 AA as the midline.In the regard to amino acid usage in CDR3 region, each sample prior used Alanine, Tyrosine,Glycine,Alanine,Aspartic acid and Serine.The amino acid usages of 107,108,109,113,114 positions were diversified but 105,106,115,116,117 positions taking conservative amino acids usages.We have found 48 unique sequences that have same IGHV, IGHJ and CDR3 AA length with the HBsAb sequences from NCBI database.Conclusion:There were almost the same characteristics of BCR CDR3 repertoire of the peripheral blood with HBsAb titer higher than 10 000 mU/ml.The 48 unique sequences provided a solid data basis for the follow-up study.

Relationship between CD4+CD25+Treg cells, Th17 cells and IL-6 and the prognosis of hepatitis B virus-related acute-on-chronic liver failure: a meta-analysis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role ofCD4+CD25+ T regulatory (Treg) cells, T helper (Th)17cells and interleukin (IL)-6 in the progression of hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) and determine their value as prognostic markers.</p><p><b>METHODS</b>The Chinese National Knowledge Infrastructure (CNKI), WanFang, Chinese Scientific Journals (VIP), PubMed, Embase and Web of Science databases were searched for English language case-control studies on the relationship between regulatory T lymphocytes and ACLF.The quality of included studies was assessed using the Newcastle-Ottawa scale. The meta-analysis was designed according to the PICOS approach recommended by the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement. RevMan software, version 5.1, was used to perform the meta-analysis.</p><p><b>RESULTS</b>Nine case-cohort studies were selected for inclusion in the metaanalysis.The results of the meta-analyses showed that the level of CD4+CD25+ Treg cells was not significantly different between patients with HBV-related ACLF and patients with chronic hepatitis B (CHB) (mean difference (MD)=0.59, 95% confidence interval (CI)-1.68, 2.85, P=0.61) nor between patients with HBVrelated ACLF and healthy controls (MD=1.12, 95% CI:-1.42, 3.66, P=0.39). Thus, it appears that ACLF patients do not have a higher level of CD4+CD25+ Treg cells than CHB patients or healthy controls. However, the ACLF patients did appear to have a significantly higher level of Th17 cells than both the CHB patients (MD=1.73, 95% CI:0.21, 3.26, P=0.03) and the healthy controls (MD=1.62, 95% CI:(0.52, 2.72, P=0.004). In addition, the ACLF patients also had significantly higher level than both the CHB patients (MD=11.69, 95%CI:1.98, 21.40, P=0.02) and the healthy controls (MD=13.17, 95% CI:1.38, 24.95, P=0.03).</p><p><b>CONCLUSION</b>CD4+CD25+ Treg cells may be an important protective factor in the progression and prognosis of HBV-related ACLF, while Thl7 cells and IL-6 may be risk factors for further progression and worsened prognosis.</p>

Analysis of styryllactones from Goniothalamus cheliensis by UPLC-Q-TOF-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the styryllactone components in Goniothalamus cheliensis Hu (Annonaceae).</p><p><b>METHOD</b>UPLC-Q-TOF-MS was used to identify the main styryllactone components in G. cheliensis. The chromatographic separation was performed on ACQU ITY UPLC BEH C18 column and eluted by actonitrile and 0.1% acetic acid in water gradiently. The mass spectrometer equipped with electrospray ionization souce was used as detector under the positive ion modes.</p><p><b>RESULT</b>Twelve styryllactons were identified based on their MS data and published literatures, and the MS fragmentation regularity of the styryllactones was also proposed.</p><p><b>CONCLUSION</b>It is an accurate and effective method to obtain the structural information of styryllactones.</p>

Analysis of the T lymphocyte receptor beta chain complementarity determining region 3 spectratyping in the peripheral blood and hepatic tissue of patients with chronic hepatitis B / 中华传染病杂志

Objective To analyse the spectral patterns of complementarity determining region 3 (CDR3) length distribution of T lymphocyte receptor beta chain variable (TRBV) gene families in infiltrating T cells of the liver tissues and the peripheral blood samples of patients with chronic hepatitis B (CHB) in order to evaluate the characteristics of T cell clonal expansion. Methods The spectral patterns drift of TRBV gene families (the monoclonal/oligoclonal TCR β T cells) in the peripheral blood and hepatic tissues from 11 cases of CHB patients were analyzed by the real-time fluorescence quantitative reverse transcription polymerase chain reaction (FQ-PCR) with DNA melting curve analysis, and abnormal rates of TRBV gene families were compared between CHB patients and healthy control. The comparison of rates was done by chi square test. Results The gene melting spectral pattern of 26 TRBV families of the 11 CHB patients, no matter in the peripheral blood or hepatic tissue, showed either a single peak or prominent melting peaks, even disappeared for certain TRBV families. The abnormal rate of TRBV gene families in the hepatic tissues was significantly higher than that in the peripheral blood samples (x2 = 23. 246, P＜0. 01). What is more interesting was that some parts of TRBV families were identical in both the peripheral blood and the hepatic tissue in certain patients. TCR BV13.1, TCR BV17 and TCR BV22 fragments were found to be restricted used in both the peripheral blood and hepatic tissue by some CHB patients. Conclusions T cells in the peripheral blood and the hepatic tissues of CHB patients can develop clonal expansion to some extent.Parts of TRBV families are restricted used in the peripheral blood and hepatic tissue in some CHB patients, which offers a foundation for further studying the common specific spectral drift patterns of TRBV CDR3 gene in CHB patients.

Association of HLA-A * 3101, B * 4001, B * 5801 and DRB1 * 1602 alleles with hemorrhagic fever with renal syndrome disease in Han population of Zunyi province / 中华微生物学和免疫学杂志

Objective To explore the association between HLA-A*31, B*40, B*58 and DRB1*16 allele polymorphisms and onset of hemorrhagic fever with renal syndrome (HFRS) in Zunyi Han population. Methods Using group study, HLA-A*31, B*40, B*58 and DRB1*16 genotyping was conducted in 100 HFRS cases and 100 controls among Han population in Zunyi area with polymerase chain reactionsequence specific primer(PCR-SSP), gene frequency (GF) and relative risk (RR) were calculated and compared. Results The results showed that the frequencies of A*3101, B*5801 and DRB1*1602 were increased in patients as compared to the healthy controls ( RR = 13. 825, x2 = 4. 296, P = 0. 038; RR =2.614,x2 =6. 133,P=0.013;RR =8.523,x2 =8. 865,P=0. 003). The frequency of B*4001 in patients with HFRS were significantly lower than that in the healthy controls( RR =0.414,x2 =6.640,P =0. 010).Conclusion These results suggest that HLA-A*3101, B*5801 and DRB1*1602 haplotypes were strongly associated with susceptibility to HFRS disease in Zunyi Han population and allele HLA-B*4001 might be associated with protection against hantaviruses infection.

The expression of monocyte chemoattractant protein-1 and CCR2 in tuberculous pleurisy / 中国综合临床

Objective To investigate the expression of monocyte chemoattractant protein-1(MCP-1)and CCR2 in peripheral blood and pleural fluid of tuberculous pleurisy patients.Methods Flow cytometry was used to detect the expression of CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients;ELISA was used to detect the content of MCP-1 in serum and pleural fluid.Results MCP-1 in surem and pleural fluid and CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients was significantly higher than that of the healthy normal controls[MCP-1:(340.8±220.8)and(9.0±3.8)ng/L,P＜0.01;CCR2(18.2±10.1)%and(6.9±3.5)%,P＜0.05];Both MCP-1 and CCR2 were detected in pleural fluid and both of them were positivley correlated(r=0.227,P＜0.05).Conclusion The expression of MCP-1 and CCR2 in peripheral blood of tuberculous pleurisy patient are significantly elevated which are significance molecule participating in the pathogenesis of tuberculous pleurisy.