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Puerarin has the anti-Alzheimer's disease (AD) activity,which can reverse nerve injury induced by Aßand inhibit neuronal apoptosis.However,its potential pharmacodynamic mechanism still needs to be further researched.The occurrence and development of AD is due to the change of multiple metabolic links in the body,which leads to the destruction of balance.Puerarin may act on multiple targets and multiple metabolic processes to achieve therapeutic purposes.Quantitative proteomic analysis provides a new choice to understand the mechanism as completely as possible.This research adopted SH-SY5Y cells induced by Aß_(1-42)to establish AD cell model,and Aßimmunofluorescence detection showed that Aßdecreased significantly after puerarin intervention.The mechanism of puerarin reversing SH-SY5Y cell injured by Aß_(1-42)was further explored by using label-free non-labeled quantitative technology and Western blot detection based on bioinformatics analysis result.The results showed that most of the differential proteins were related to biological processes such as cellular component organization or biogenesis,cellular component organization and cellular component biogenesis,and they mainly participated in the top ten pathways of P value such as pathogenic Escherichia coli infection,m TOR signaling pathway,regulation of autophagy,regulation of actin cytoskeleton,spliceosome,hepatocellular carcinoma,tight junction,non-small cell lung cancer,apoptosis and gap junction.Annexin V/PI flow cytometry and TUNEL were used to detect apoptosis,and the results showed that Aßdecreased significantly and the rate of apoptosis decreased significantly after puerarin intervention.Western blot analysis found that the protein expression level of autophagy related protein LC3â ¡was up-regulated after Aßinduction,and the degree of this up-regulation was further enhanced in puerarin intervention group.The trend of the ratio of LC3â ¡/LC3â among groups was the same as the protein expression level of LC3â ¡ï¼the protein expression level of p62 in the control group,AD model group and puerarin intervention group decreased successively.Protein interaction network analysis showed that CAP1 was correlated with TUBA1B,HSP90AB2P,DNM1L,TUBA1A and ERK1/2,and the correlation between CAP1 and ERK1/2 was the highest among them.Western blot showed that the expressions of p-ERK1/2,Bax and CAP1 were significantly down-regulated and the protein expression level of Bcl-2 was significantly up-regulated after puerarin intervention.Therefore,puerarin might improve the SH-SY5Y cells injured by Aß_(1-42)through the interaction of multiple biological processes and pathways in cells multiple locations,and CAP1 might play an important role among them.
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Carcinoma de Pulmón de Células no Pequeñas , Isoflavonas , Neoplasias Pulmonares , Péptidos beta-Amiloides , Apoptosis , Línea Celular Tumoral , Humanos , Isoflavonas/farmacología , ProteómicaRESUMEN
Glutamine, like glucose, is a major nutrient consumed by cancer cells, yet these cells undergo glutamine starvation in the cores of tumors, forcing them to evolve adaptive metabolic responses. Pharmacologically targeting glutamine metabolism or withdrawal has been exploited for therapeutic purposes, but does not always induce cancer cell death. The mechanism by which cancer cells adapt to resist glutamine starvation in cisplatin-resistant non-small-cell lung cancer (NSCLC) also remains uncertain. Here, we report the potential metabolic vulnerabilities of A549/DDP (drug-resistant human lung adenocarcinoma cell lines) cells, which were more easily killed by the iron chelator deferoxamine (DFO) during glutamine deprivation than their parental cisplatin-sensitive A549 cells. We demonstrate that phenotype resistance to cisplatin is accompanied by adaptive responses during glutamine deprivation partly via higher levels of autophagic activity and apoptosis resistance characteristics. Moreover, this adaptation could be explained by sustained glucose instead of glutamine-dominant complex II-dependent oxidative phosphorylation (OXPHOS). Further investigation revealed that cisplatin-resistant cells sustain OXPHOS partly via iron metabolism reprogramming during glutamine deprivation. This reprogramming might be responsible for mitochondrial iron-sulfur [Fe-S] cluster biogenesis, which has become an "Achilles' heel," rendering cancer cells vulnerable to DFO-induced autophagic cell death and apoptosis through c-Jun N-terminal kinase (JNK) signaling. Finally, in vivo studies using xenograft mouse models also confirmed the growth-slowing effect of DFO. In summary, we have elucidated the adaptive responses of cisplatin-resistant NSCLC cells, which balanced stability and plasticity to overcome metabolic reprogramming and permitted them to survive under stress induced by chemotherapy or glutamine starvation. In addition, for the first time, we show that suppressing the growth of cisplatin-resistant NSCLC cells via iron chelator-induced autophagic cell death and apoptosis was possible with DFO treatment. These findings provide a solid basis for targeting mitochondria iron metabolism in cisplatin-resistant NSCLC for therapeutic purposes, and it is plausible to consider that DFO facilitates in the improvement of treatment responses in cisplatin-resistant NSCLC patients.
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This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-ß (TGF-ß), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.
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Arteriosclerosis Obliterante/patología , Autofagia , Polaridad Celular , Macrófagos/citología , Amputados , Arginasa/metabolismo , Aterosclerosis/patología , Quimiocina CCL2/metabolismo , Células Espumosas/citología , Humanos , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Interleucina-6/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenotipo , Factor de Transcripción STAT6/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia ArribaRESUMEN
Lupus erythematosus panniculitis (LEP) is a variant of chronic cutaneous lupus erythematosus (CCLE). Reported cases of LEP lesions before the diagnosis of systemic lupus erythematosus (SLE) were very rare; only 9 cases have been reported, to the best of our knowledge. We now describe the case of a 19-year-old male patient, with an overall review of the English literature. In the earliest stage of the present case, nodules and ulcers involved his left leg and face, with no other accompanied symptoms. The skin lesions disappeared after treatment with methylprednisolone, 16âmg/d for 1 month. Seven months after discontinuing methylprednisolone, the cutaneous nodules and ulcers on his back recurred and were accompanied by fever, hair loss, and polyarthritis. Blood tests revealed leucopenia, positive antinuclear antibody and Smith antibody, and proteinuria. Histopathological findings were most consistent with LEP. This was followed sequentially by the diagnosis of SLE. The patient improved again after treatment with methylprednisolone and cyclophosphamide.Patients with LEP should have regular follow-ups because the development of SLE is possible. Early diagnosis and proper treatment is pivotal to improve the prognosis of such patients.
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Lupus Eritematoso Sistémico/complicaciones , Paniculitis de Lupus Eritematoso/etiología , Piel/patología , Biopsia , Diagnóstico Diferencial , Humanos , Lupus Eritematoso Sistémico/diagnóstico , Masculino , Paniculitis de Lupus Eritematoso/diagnóstico , Recurrencia , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
AIM OF STUDY: Tanshinone IIA is an active component of the traditional Chinese medicine. This study aimed at investigating the mechanism of tanshinone IIA on anti-atherosclerosis, which may be because of that Tanshinone IIA can affect the HDL subfractions distribution and then regulate reverse cholesterol transport. MATERIALS AND METHODS: A model of hyperlipidaemia in rats was used. Tanshinone IIA was given daily after hyperlipidaemia. In vivo, lipid deposition and morphological changes in liver were analyzed; HDL subfractions and lipid level in serum as well as in liver were measured; the expression of genes related to cholesterol intake in liver and peritoneal macrophage cholesterol efflux were evaluated. In vitro, HepG2 cells and THP-1 cells were pretreated with tanshinone IIA and subsequently with ox-LDL to evaluate the total cholesterol and the expression of related genes. RESULTS: Tanshinone IIA reduced the lipid deposition in liver. Moreover, it did not affect the serum lipid levels but reduced the levels of HDL middle subfractions and increased the levels of HDL large subfractions. Furthermore, tanshinone IIA could regulate the expressions of CYP7A1, LDL-R, SREBP2 and LCAT in the liver as well as the ABCA1 and CD36 in macrophage cells which is involving in the cholesterol intake and efflux respectively. It could reduce lipid accumulation caused by ox-LDL induction, and that also regulate the expressions of LDL-R, HMGCR and SREBP2 in HepG2 and ABCA1, CD36 in THP-1 cells. CONCLUSION: A novel finding that tanshinone IIA was not reduce the serum lipid level but affects the HDL subfractions distribution and thereby regulating the intake and efflux of cholesterol.
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Abietanos/administración & dosificación , HDL-Colesterol/metabolismo , Hiperlipidemias/metabolismo , Metabolismo de los Lípidos , Lipoproteínas HDL/metabolismo , Hígado/metabolismo , Animales , Transporte Biológico Activo , Hiperlipidemias/tratamiento farmacológico , Lípidos/sangre , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Both shingles and psoriasis are common cutaneous diseases. About 25% of the psoriatic patients develop Koebner phenomenon (KP) after various injuries, and in rare instance, KP may occur at the site of healed or healing shingles.We report a 30-year-old man with 7-month history of scalp psoriasis who developed KP at the areas of developing shingles. Cutaneous examination revealed scaly erythematous papules and plaques located on the scalp and forehead, and groups of clustered erythematous papules with silver scales in the dermatome distributed on the right side of chest wall the prior herpes zoster lesions involved. After removal of the scales on the papules, underlying bleeding points were present.The lesions on chest had good response to anti-psoriatic therapies, as the lesions on scalp did. After a year of follow-up, recurrent psoriasis occurred, but the lesions were located only on the scalp, and the areas of prior occurrence of shingles, because of which we considered diagnosis of recurrent psoriasis rather than relapsing KP for the chest lesions.Not only the healing and healed shingles can trigger KP in psoriasis, but also the developing shingles can cause psoriatic KP at the site of herpes zoster lesions.
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Herpes Zóster/complicaciones , Psoriasis/complicaciones , Adulto , Herpes Zóster/patología , Humanos , Masculino , Psoriasis/patología , Piel/patologíaRESUMEN
OBJECTIVE: To explore the effect of arteriosclerosis obliterans (ASO) blood stasis syndrome (BSS) serum on vascular endothelial cell injury and to study the regulation of Taohong Siwu Decoction (TSD) on it. METHODS: Umbilical vein endothelial cell culture system was established. The serum endothelial cell injury model with ASO BSS was prepared. Low, medium, and high concentrations TSD containing serums were respectively added. The endothelial cell proliferation activity was observed by MTT method. Ultrastructures of endothelial cells were observed under transmission electron microscope. Changes of intracellular calcium ion concentration and the cytoskeleton were observed under laser confocal microscope. Contents of ET, NO, and transforming growth factor beta1 (TGF-beta1) in endothelial cell culture supernatant were detected by ELISA. RESULTS: In ASO BSS serum group endothelial cell proliferation activities decreased, the cell structure was obviously destroyed, calcium ion concentration increased, contents of ET, NO and TGF-beta1 increased significantly (P < 0.01), and ET/NO ratio was imbalanced. After incubating with TSD drug containing serum, endothelial cell proliferation activities and injured cell structures were obviously improved; ET, NO and TGF-beta1 levels decreased (P < 0.05, P < 0.01), ET/NO ratios approximated to the normal level. CONCLUSION: The main mechanism of TSD for treating ASO ASS lied in improving injured vascular endothelial cells and endocrine disorder.
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Arteriosclerosis Obliterante , Medicamentos Herbarios Chinos/uso terapéutico , Medicina Tradicional China , Proliferación Celular , Células Endoteliales , Humanos , Suero , Factor de Crecimiento Transformador beta1/metabolismo , Venas UmbilicalesRESUMEN
OBJECTIVE: To discuss the effect of Taohong Siwu Decoction (TSD) in regulating functions of endothelial cells and treating arteriosclerosis obliterans (ASO). METHODS: The ASO model was prepared by using high-fat diet plus intimal injury. They were randomly divided into the model group (n = 10), the normal control group (n = 9), the low dose TSD group (group A, n = 12), the middle dose TSD group (group B, n = 10), and the high dose TSD group (group C, n = 9). Eight weeks after modeling, the limb blood perfusion was observed using laser Doppler flowmetry. The arterial morphology was observed using light microscope and transmission electron microscope. The number of circulating endothelial cells (CECs) was determined using Percoll density gradient centrifugation method. Serum levels of TNF-alpha, IL-1, ET-1, and NO were detected using double antibody sandwich assay of enzyme linked immunosorbent assay (ELISA). RESULTS: The ASO rat model was successfully established. Blood lipids levels significantly increased, the blood perfusion of left hind limbs significantly decreased, the number of CECs in the peripheral blood significantly increased, the arterial lumen was irregularly narrowed, the ultra-structure of vessel walls was damaged, serum levels of TNF-alpha, IL-1, and ET-1 significantly increased, and the serum level of NO significantly decreased in the model group, showing statistical difference when compared with the normal control group (P < 0.01). Compared with the model group, significant improvement in the aforesaid indices was shown in group B and C (P < 0.05, P < 0.01). CONCLUSIONS: The injury and abnormal functions of endothelial cells is an important pathological process of ASO. As an effective recipe for treating ASO, TSD could protect vascular endothelial cells and improve the secretion function of vascular endothelial cells.
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Arteriosclerosis Obliterante/sangre , Medicamentos Herbarios Chinos/farmacología , Células Endoteliales/metabolismo , Endotelio Vascular/citología , Animales , Arteriosclerosis Obliterante/tratamiento farmacológico , Dieta Alta en Grasa/efectos adversos , Medicamentos Herbarios Chinos/uso terapéutico , Endotelina-1/sangre , Interleucina-1/sangre , Masculino , Óxido Nítrico/sangre , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/sangreRESUMEN
BACKGROUND: Cultivated bananas are large, vegetatively-propagated members of the genus Musa. More than 1,000 cultivars are grown worldwide and they are major economic and food resources in numerous developing countries. It has been suggested that cultivated bananas originated from the islands of Southeast Asia (ISEA) and have been developed through complex geodomestication pathways. However, the maternal and parental donors of most cultivars are unknown, and the pattern of nucleotide diversity in domesticated banana has not been fully resolved. METHODOLOGY/PRINCIPAL FINDINGS: We studied the genetics of 16 cultivated and 18 wild Musa accessions using two single-copy nuclear (granule-bound starch synthase I, GBSS I, also known as Waxy, and alcohol dehydrogenase 1, Adh1) and two chloroplast (maturase K, matK, and the trnL-F gene cluster) genes. The results of phylogenetic analyses showed that all A-genome haplotypes of cultivated bananas were grouped together with those of ISEA subspecies of M. acuminata (A-genome). Similarly, the B- and S-genome haplotypes of cultivated bananas clustered with the wild species M. balbisiana (B-genome) and M. schizocarpa (S-genome), respectively. Notably, it has been shown that distinct haplotypes of each cultivar (A-genome group) were nested together to different ISEA subspecies M. acuminata. Analyses of nucleotide polymorphism in the Waxy and Adh1 genes revealed that, in comparison to the wild relatives, cultivated banana exhibited slightly lower nucleotide diversity both across all sites and specifically at silent sites. However, dramatically reduced nucleotide diversity was found at nonsynonymous sites for cultivated bananas. CONCLUSIONS/SIGNIFICANCE: Our study not only confirmed the origin of cultivated banana as arising from multiple intra- and inter-specific hybridization events, but also showed that cultivated banana may have not suffered a severe genetic bottleneck during the domestication process. Importantly, our findings suggested that multiple maternal origins and a reduction in nucleotide diversity at nonsynonymous sites are general attributes of cultivated bananas.
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Cloroplastos/genética , Genes de Plantas , Musa/genética , Asia Sudoriental , Frecuencia de los Genes , Variación Genética , Genoma de Planta , Haplotipos , Hibridación Genética , Islas , Filogenia , Proteínas de Plantas/genética , Polimorfismo Genético , Almidón Sintasa/genéticaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Salvia Miltiorrhiza Bunge (also known as herb Danshen in Chinese) is a widely used Chinese herbal medicine. Tanshinone IIA (TSN IIA) is considered to be the most important bioactive ingredient in Danshen and exhibits an anti-atherosclerotic activity. AIM OF STUDY: To evaluate the protective effect of TSN IIA on the human endothelial EA.hy926 cells injured by hydrogen peroxide in vitro and its possible mechanism. MATERIALS AND METHODS: The EA.hy926 cells were incubated for 24h with different concentrations of TSN IIA (5, 10 and 20 µg/µL ) or DMEM. Subsequently, cells were treated with 300 µmol/L H(2)O(2) for another 4h. Then, the percentage of cell viability was evaluated by 3-(4, 5-di-methylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The apoptosis of EA.hy926 cells was detected by flow cytometry with AnnexinV-FITC/PI double staining and laser scanning spectral confocal technique. The generation of intracellular reactive oxygen species (ROS) generation was analyzed by flow cytometry. The mRNA expressions of caspase-3, Bcl-2 and Bax were tested by real time-reverse transcription polymerase chain reaction (real time RT-PCR). The protein expression of Bcl-2 and Bax was determined by Western blotting. MDA levels, NO production, LDH leakage, and SOD as well as caspase-3 activities were also measured using standard methods. RESULTS: Loss of cell viability and excessive cell apoptosis were observed in EA.hy926 cells after 4h of challenge with H(2)O(2) (300 µmol/L). However, cell apoptosis was attenuated in different concentrations of TSN IIA (5, 10 and 20 µg/µL) pretreated cells. Furthermore, TSN IIA markedly inhibited the elevation of ROS evoked by H(2)O(2). Real time RT-PCR and Western blotting analysis showed that TSN IIA significantly decreased the expressions of pro-apoptotic proteins (Bax and caspase-3) while significantly increased the expression of anti-apoptotic protein Bcl-2, and resulted in obvious reduction of Bax/Bcl-2 ratio in EA.hy926 cells induced by H(2)O(2). CONCLUSION: These observations provide preliminary evidence that TSN IIA protects EA.hy926 cells against H(2)O(2) damage, which is mainly associated with the ROS generation, followed by the imbalance of the Bax/Bcl-2 ratio, and caspase-3 activation leading to apoptosis.
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Abietanos/farmacología , Apoptosis/efectos de los fármacos , Aterosclerosis/metabolismo , Medicamentos Herbarios Chinos/farmacología , Células Endoteliales/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Fenantrolinas/farmacología , Salvia miltiorrhiza/química , Abietanos/uso terapéutico , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Aterosclerosis/prevención & control , Caspasa 3/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/uso terapéutico , Células Endoteliales/metabolismo , Humanos , Peróxido de Hidrógeno , Fenantrolinas/uso terapéutico , Fitoterapia , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
AIM: To observe the influence of tanshinone II A on the expression of toll like receptor 4(TLR4) and tumor necrosis factor-α(TNF-α) in cultured human umbilical veins endothelial cells EA.hy926 induced by LPS, and to investigate the molecular mechanism of tanshinone II A against atherosclerosis(AS). METHODS: The EA.hy926 cells were cultured in vitro. The mRNA and protein expression of TLR4 were tested by RT-PCR and immunocytochemistry technique respectively. The mRNA and protein expression of TNF-α were detected by RT-PCR and ELISA technique respectively. RESULTS: The expression of TLR4 and TNF-α were both increased compared to normal group after stimulation of LPS(P<0.05). The expression of TLR4 and TNF-α in tanshinone II A group were obviously inhibited compared to stimulation group(P<0.05). CONCLUSION: The molecular mechanism of tanshinone II A against AS is probably to inhibit the expression of TLR4 and TNF-α.
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Abietanos/farmacología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Lipopolisacáridos/farmacología , Receptor Toll-Like 4/genética , Factor de Necrosis Tumoral alfa/genética , Abietanos/uso terapéutico , Aterosclerosis/tratamiento farmacológico , Línea Celular , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Interdigital ulcer is an exceptionally rare condition while erosio interdigitalis blastomycetica is common for candidiasis. Four Chinese patients with Candida interdigital ulcers were reported. The exudates were examined directly and cultured for fungi. Skin biopsies were stained with haematoxylin-eosin and periodic acid Schiff. There were a man and three women (age range: 34-56 years) who presented with 1- to 3-month history of chronic cutaneous ulcer on the interdigital web of hand or foot. The lesions were located on hand for one woman, and on the left foot for the rest. The patients had poor response to the previous treatment of topical steroids and oral antimicrobials. Candida albicans was isolated from a man and two women, Candida tropicalis from another woman. Biopsy specimens revealed yeast and mycelium as well as inflammatory infiltrate in necrotic tissue in two patients; only inflammatory cells in the other two. The patients had complete remission with oral itraconazole and topical bifonazole cream therapy for 3- to 5-week. Candida species may cause interdigital ulcer on hand or foot. Oral itraconazole and topical bifonazole may be an optional therapy for such an ulcer.