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1.
Zhongguo Zhong Yao Za Zhi ; 47(14): 3781-3787, 2022 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-35850835

RESUMO

Since the current identification method for Paeoniae Radix Alba is complex in operation and long time-consuming with high requirements for technicians, the present study employed Heracles NEO ultra-fast gas phase electronic nose(E-nose) technology to identify raw and sulfur-fumigated Paeoniae Radix Alba decoction pieces in order to establish a rapid identification method for sulfur-fumigated Paeoniae Radix Alba. The odors of raw Paeoniae Radix Alba and its sulfur-fumigated products were analyzed by Heracles NEO ultra-fast gas phase E-nose to obtain the odor chromatographic information. The chemometric model was established, and the data were processed by principal component analysis(PCA), discriminant function analysis(DFA), soft independent modeling of class analogy(SIMCA), and partial least squares discriminant analysis(PLS-DA). The differential compounds of raw and sulfur-fumigated samples were qualitatively analyzed based on the Kovats retention index and Arochembase. As revealed by the comparison of gas chromatograms of raw and sulfur-fumigated Paeoniae Radix Alba, the heights of several peaks in the chromatograms before and after sulfur fumigation changed significantly. The peak(No.8) produced by ethylbenzene disappeared completely due to sulfonation reaction in the process of sulfur fumigation, indicating that ethylbenzene may be the key component in the identification of Paeoniae Radix Alba and its sulfur-fumigated products. In PCA, DFA, SIMCA, and PLS-DA models, the two types of samples were separated into two different regions, indicating that the established models can clearly distinguish between raw and sulfur-fumigated Paeoniae Radix Alba. The results showed that Heracles NEO ultra-fast gas phase E-nose technology could realize the rapid identification of raw and sulfur-fumigated Paeoniae Radix Alba, which provides a new method and idea for the rapid identification of sulfur-fumigated Chinese medicine.


Assuntos
Medicamentos de Ervas Chinesas , Paeonia , Medicamentos de Ervas Chinesas/química , Nariz Eletrônico , Fumigação/métodos , Paeonia/química , Extratos Vegetais , Enxofre/química
2.
Microbiol Spectr ; 10(4): e0026022, 2022 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-35735985

RESUMO

Thuja sutchuenensis Franch. is an endangered species in southwest China, distributed sporadically in mountainous areas. Soil property and soil fungal community play a crucial role in plant growth and survival. Nevertheless, understanding soil properties and the soil fungal community in the areas where T. sutchuenensis is distributed is extremely limited. Hence, this study collected a total of 180 soil samples from five altitudinal distribution areas (altitudinal gradients) and three vertical depths throughout four horizontal distances from the base of each tree. The results found that altitudinal gradients and vertical depths altered soil properties, including pH, organic matter content, water content, total nitrogen, phosphorus, and potassium, and available nitrogen, phosphorus, and potassium. The fungal alpha diversity indexes (Chao1 and Shannon) and beta diversity were dramatically decreased with elevation. In addition, high altitudes (2,119 m) harbored the highest relative abundance of ectomycorrhizal fungi (27.57%) and the lowest relative abundance of plant-pathogenic fungi (1.81%). Meanwhile, we identified a series of fungal communities, such as Tomentella, Piloderma, Cortinarius, Sebacina, and Boletaceae, that play an essential role in the survival of T. sutchuenensis. The correlation analysis and random forest model identified that water content and total phosphorus showed strong relationships with fungal characteristics and were the primary variables for Zygomycota and Rozellomycota. Collectively, the findings of this integrated analysis provide profound insights into understanding the contrasting responses of T. sutchuenensis soil fungal communities and provide a theoretical basis for T. sutchuenensis habitat restoration and species conservation from multispatial perspectives. IMPORTANCE The present study highlights the importance of fungal communities in an endangered plant, T. sutchuenensis. Comparative analysis of soil samples in nearly all extant T. sutchuenensis populations identified that soil properties, especially soil nutrients, might play critical roles in the survival of T. sutchuenensis. Our findings prove that a series of fungal communities (e.g., Tomentella, Piloderma, and Cortinarius) could be key indicators for T. sutchuenensis survival. In addition, this is the first time that large-scale soil property and fungal community investigations have been carried out in southwest China, offering important values for exploring the distribution pattern of regional soil microorganisms. Collectively, our findings display a holistic picture of soil microbiome and environmental factors associated with T. sutchuenensis.


Assuntos
Basidiomycota , Micobioma , Micorrizas , Thuja , Traqueófitas , Fungos , Nitrogênio , Fósforo , Plantas , Potássio , Solo/química , Microbiologia do Solo , Água
3.
J Trace Elem Med Biol ; 71: 126960, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35219977

RESUMO

BACKGROUND: Selenium concentration of one spot urine sample cannot reflect selenium status during whole pregnancy. Studies on variations of urinary selenium concentration at different stages of pregnancy are limited. AIM: To assess variations of urinary selenium concentrations during three trimesters of pregnancy and to explore the influencing factors. METHODS: This study included 2613 pregnant women from a birth cohort study (Wuhan, China) enrolled between October 2014 and October 2016. Selenium concentrations of urine samples collected at three trimesters were measured. We used a generalized linear mixed effects model to observe the changes in urinary selenium concentration during pregnancy and its influencing factors. RESULTS: The median value (range) of gestational weeks at urine sample collection was 11 (9-14), 24 (15-27), and 34 (28-41) respectively. Urine selenium concentration varied across trimesters (geometric mean: 16.34, 17.65 and 18.83 µg/g creatinine, respectively), with an upward trend (ß = -0.145, 95%CI: -0.164, -0.126) (ß = -0.066, 95%CI: -0.083, -0.048). The concentrations of urinary selenium increased with the increasing of educational level [ß (95%CI): ≤ 9 years = -0.105 (-0.163, -0.047); 10-12 years = -0.086(-0.126, -0.047); > 12 years = reference]. Pregnant women who rarely or only took multivitamins in the first trimester [ß (95% CI): rarely = -0.076 (-0.144, -0.007); only in the first three months of pregnancy = -0.104 (-0.170, -0.038); always = reference], or were not exposed to passive smoking during pregnancy(ߠ= -0.093, 95% CI: - 0.173, - 0.014), or exercised three to four days per week for the first three months before delivery(ߠ= -0.074, 95% CI: - 0.140, - 0.008) had lower urinary selenium concentrations. Pregnant women who took calcium supplements after the third month of pregnancy had higher urinary selenium levels(ߠ= 0.114, 95% CI: 0.059, 0.169). CONCLUSIONS: Urine selenium concentrations of pregnant women showed an upward trend across trimesters. Educational level, calcium or multivitamin supplementation, passive smoking, and physical exercise might impact urine selenium levels.


Assuntos
Selênio , Poluição por Fumaça de Tabaco , Gravidez , Feminino , Humanos , Estudos Longitudinais , Estudos de Coortes , Cálcio , Trimestres da Gravidez
4.
Biomed Res Int ; 2021: 3664564, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34853789

RESUMO

Tumor necrosis factor-α is a common cytokine that increases in inflammatory processes, slows the differentiation of bone formation, and induces osteodystrophy in the long-term inflammatory microenvironment. Our previous study confirmed that the Elongation protein 2 (ELP2) plays a significant role in osteogenesis and osteogenic differentiation, which is considered a drug discovery target in diseases related to bone formation and differentiation. In this study, we applied an in silico virtual screening method to select molecules that bind to the ELP2 protein from a chemical drug molecule library and obtained 95 candidates. Then, we included 11 candidates by observing the docking patterns and the noncovalent bonds. The binding affinity of the ELP2 protein with the candidate compounds was examined by SPR analysis, and 5 out of 11 compounds performed good binding affinity to the mouse ELP2 protein. After in vitro cell differentiation assay, candidates 2# and 5# were shown to reduce differentiation inhibition after tumor necrosis factor-α stimulation, allowing further optimization and development for potential clinical treatment of inflammation-mediated orthopedic diseases.


Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Osteogênese/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Células 3T3 , Animais , Calcificação Fisiológica/efeitos dos fármacos , Calcificação Fisiológica/fisiologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Linhagem Celular , Bases de Dados de Produtos Farmacêuticos , Avaliação Pré-Clínica de Medicamentos , Marcadores Genéticos , Técnicas In Vitro , Peptídeos e Proteínas de Sinalização Intracelular/química , Ligantes , Camundongos , Modelos Moleculares , Simulação de Acoplamento Molecular , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteogênese/genética , Osteogênese/fisiologia , Ligação Proteica , Relação Estrutura-Atividade , Ressonância de Plasmônio de Superfície , Interface Usuário-Computador
5.
Artigo em Inglês | MEDLINE | ID: mdl-34217844

RESUMO

The present study investigated the effects of dietary Astragalus Propinquus schischkin polysaccharides on growth, immune responses, antioxidants responses and inflammation-related genes expression in Channa argus. Channa argus were randomly divided into 5 groups and fed 5 levels diets of A. propinquus schischkin polysaccharides (0, 250, 500, 1000 and 2000 mg/kg) for 56 days. The results showed, dietary A. propinquus schischkin polysaccharides addition can increase the final body weight, weight gain and specific growth rate, decrease the feed conversion ratio of Channa argus. And dietary A. propinquus schischkin polysaccharides supplementation can increase the levels of serum superoxide dismutase, catalase, glutathione peroxidase, lysozyme, complement 3, complement 4, immunoglobulin M and alkaline phosphatase, decrease the levels of serum malondialdehyde, cortisol, aspartate aminotransferase and glutamic-pyruvic transaminase. Furthermore, dietary A. propinquus schischkin polysaccharides can decrease the gene expression levels of interleukin-1ß, interleukin-, tumor necrosis factor-α and nuclear factor-κB, increase the gene expression levels of glucocorticoid receptor in liver, spleen, kidney, intestine. To sum up, dietary A. propinquus schischkin polysaccharides can accelerate growth, enhance immune responses and antioxidants responses, regulate inflammation-related genes expression in Channa argus and the optimum amount is 1000 mg/kg.


Assuntos
Astragalus propinquus/química , Proteínas de Peixes/genética , Peixes/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Polissacarídeos/farmacologia , Ração Animal , Animais , Antioxidantes/metabolismo , Aquicultura , Peso Corporal/efeitos dos fármacos , Suplementos Nutricionais , Peixes/imunologia , Imunoglobulina M/sangue , Inflamação/dietoterapia , Inflamação/genética , Inflamação/veterinária , Polissacarídeos/administração & dosagem
6.
Zhongguo Zhong Yao Za Zhi ; 45(21): 5209-5218, 2020 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-33350237

RESUMO

Licorice has long been regarded as one of the most popular herbs, with a very wide clinical application range. Whether being used alone or as an ingredient in prescription, it has an important role which cannot be ignored. However, the efficacy and chemical constituents of licorice will change after honey-processing. Therefore, it is necessary to find quality markers before and after honey-processing to lay the foundation for a comprehensive evaluation of the differences between raw and processed licorice pieces. HPLC-DAD was employed to establish fingerprints of raw and processed licorice. Multivariate statistical analysis methods including principal component analysis(PCA) and orthogonal partial least squares discrimination analysis(OPLS-DA) were applied to screen out the differential components before and after processing of licorice. Based on network pharmacology, the targets and pathways corresponding to the differential components were analyzed with databases such as Swiss Target Prediction and Metascape, and the "component-target-pathway" diagram was constructed with Cytoscape 3.6.0 software to predict the potential quality markers. A total of 17 common peaks were successfully identified in the established fingerprint, and seven differential components were selected as potential quality markers(licoricesaponin G2, glycyrrhizic acid, liquiritigenin, liquiritin, isoliquiritin, liquiritin apioside and isoliquiritigenin). The HPLC fingerprint method proposed in this study was efficient and feasible. The above seven differential chemical components screened out as potential quality markers of licorice can help to improve and promote the overall quality. These researches offer more sufficient theoretical basis for scientific application of licorice and its corresponding products.


Assuntos
Medicamentos de Ervas Chinesas , Glycyrrhiza , Mel , Cromatografia Líquida de Alta Pressão , Ácido Glicirrízico/análise , Mel/análise
7.
Zhongguo Zhong Yao Za Zhi ; (24): 5209-5218, 2020.
Artigo em Chinês | WPRIM | ID: wpr-878807

RESUMO

Licorice has long been regarded as one of the most popular herbs, with a very wide clinical application range. Whether being used alone or as an ingredient in prescription, it has an important role which cannot be ignored. However, the efficacy and chemical constituents of licorice will change after honey-processing. Therefore, it is necessary to find quality markers before and after honey-processing to lay the foundation for a comprehensive evaluation of the differences between raw and processed licorice pieces. HPLC-DAD was employed to establish fingerprints of raw and processed licorice. Multivariate statistical analysis methods including principal component analysis(PCA) and orthogonal partial least squares discrimination analysis(OPLS-DA) were applied to screen out the differential components before and after processing of licorice. Based on network pharmacology, the targets and pathways corresponding to the differential components were analyzed with databases such as Swiss Target Prediction and Metascape, and the "component-target-pathway" diagram was constructed with Cytoscape 3.6.0 software to predict the potential quality markers. A total of 17 common peaks were successfully identified in the established fingerprint, and seven differential components were selected as potential quality markers(licoricesaponin G2, glycyrrhizic acid, liquiritigenin, liquiritin, isoliquiritin, liquiritin apioside and isoliquiritigenin). The HPLC fingerprint method proposed in this study was efficient and feasible. The above seven differential chemical components screened out as potential quality markers of licorice can help to improve and promote the overall quality. These researches offer more sufficient theoretical basis for scientific application of licorice and its corresponding products.


Assuntos
Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas , Glycyrrhiza , Ácido Glicirrízico/análise , Mel/análise
8.
Biol Trace Elem Res ; 189(1): 180-185, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30069692

RESUMO

Zinc (Zn) highly concentrates in the brain and plays a key role in memory formation and learning processes. Zn deficiency results in cognitive impairments, memory deficits, alterations of neuropsychological behavior, and motor development. Although Zn-enriched yeast (ZnY) is widely used for dietary fortification and supplementation of Zn, the effect of ZnY on cognition still remains unclear. The purpose of the study was to investigate the effect of ZnY on behavior in Zn-deficient and Zn-sufficient rats. Three-week-old rats were fed low Zn diets for 145 days to establish Zn-deficient rats. ZnY was orally administered to Zn-deficient rats at three dose levels of 1, 2, and 4 mg Zn/kg/day for 55 days. Rat appearance, body weight, plasma and brain Zn, Morris water maze test, and step-through passive avoidance test were observed. Compared to Zn-sufficient rats, body weight gain, plasma zinc level, resident time, and step-through time in Zn-deficient rats were significantly lower. Zn deficiency impaired functions of learning and memory, while ZnY as a plausible therapeutic intervention alleviated the cognitive impairments caused by Zn deficiency.


Assuntos
Fermento Seco , Zinco/farmacologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Suplementos Nutricionais , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/efeitos dos fármacos , Transtornos da Memória/sangue , Transtornos da Memória/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Zinco/sangue
9.
Anat Rec (Hoboken) ; 300(10): 1858-1864, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28598552

RESUMO

Diabetes-induced neuropathic pain (DNP) substantially influences people's life qualities. Hyperglycemia-induced excess free radicals have been considered as the most critical mechanisms underlying DNP. As an unsaturated aldehyde and a reactive product of lipid peroxidation, acrolein plays critical roles in diabetic nephropathy and inflammatory pain. We sought to determine whether acrolein is involved in DNP in this study. Diabetes was induced by a single intraperitoneal (i.p.) injection of 60 mg/kg streptozotocin (STZ). An acrolein scavenger hydralazine (5 mg/kg) was administered through a daily injection for 4 weeks, starting immediately within 30 min after STZ injection. Western blot showed that hydralazine could effectively inhibit STZ-induced upregulation of acrolein in the spinal dorsal horn on day 7-28 after STZ injection. Behavioral tests showed that STZ injection induced significant mechanical allodynia and thermal hyperalgesia, which could be alleviated by hydralazine. Immunofluorescent histochemistry and Western blot showed that STZ induced significant microglial activation. ELISA data indicated upregulation of inflammatory cytokines IL-1ß and TNF-α expression in the spinal dorsal horn. Furthermore, hydralazine effectively attenuated microglial activation and expression of inflammatory mediators. Our data indicate that acrolein might be involved in the development of neuroinflammation and behavioral consequences of DNP. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 300:1858-1864, 2017. © 2017 Wiley Periodicals, Inc.


Assuntos
Acroleína/metabolismo , Nefropatias Diabéticas/etiologia , Hidralazina/uso terapêutico , Corno Dorsal da Medula Espinal/metabolismo , Vasodilatadores/uso terapêutico , Animais , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/prevenção & controle , Avaliação Pré-Clínica de Medicamentos , Hidralazina/farmacologia , Masculino , Ratos Sprague-Dawley , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Estreptozocina , Vasodilatadores/farmacologia
10.
J Asian Nat Prod Res ; 12(7): 576-81, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20628936

RESUMO

Three new lupane-type triterpenes, 3 alpha-O-trans-feruloylbetulinic acid (1), 3 alpha-O-trans-coumaroylbetulinic acid (2) and 3beta-O-cis-feruloylbetulin (3), together with 10 known triterpenes (4-13), were isolated from the aerial parts of the mangrove plant Ceriops tagal. The structures of the three new compounds were established by means of spectroscopic data analyses and chemical methods.


Assuntos
Medicamentos de Ervas Chinesas/isolamento & purificação , Rhizophoraceae/química , Triterpenos/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Triterpenos/química
11.
J Agric Food Chem ; 58(9): 5368-73, 2010 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-20297841

RESUMO

The phytochemical profiles of Morus nigra roots and twigs were compared by HPLC with those of the old and young twigs of Morus alba which are known to contain oxyresveratrol and mulberroside A as major components. It was found that M. nigra root extract contains some unknown natural products with potential tyrosinase inhibitory activity. The extract (95% ethanol) of the roots of M. nigra was further investigated in this study. One new compound, 5'-geranyl-5,7,2',4'-tetrahydroxyflavone, and twenty-eight known phenolic compounds were isolated. Their structures were identified by mass spectrometry and NMR spectroscopy. Nine compounds, 5'-geranyl-5,7,2',4'-tetrahydroxyflavone, steppogenin-7-O-beta-D-glucoside, 2,4,2',4'-tetrahydroxychalcone, moracin N, kuwanon H, mulberrofuran G, morachalcone A, oxyresveratrol-3'-O-beta-D-glucopyranoside and oxyresveratrol-2-O-beta-D-glucopyranoside, showed better tyrosinase inhibitory activities than kojic acid. It was noteworthy that the IC(50) values of 2,4,2',4'-tetrahydroxychalcone and morachalcone A were 757-fold and 328-fold lower than that of kojic acid, respectively, suggesting a great potential for their development as effective natural tyrosinase inhibitors.


Assuntos
Inibidores Enzimáticos/farmacologia , Monofenol Mono-Oxigenase/antagonistas & inibidores , Morus/química , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Relação Estrutura-Atividade
12.
Nat Prod Commun ; 5(1): 9-12, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20184010

RESUMO

A new dolabrane-type diterpene named tagalsin O 1, together with six known analogues 2-7, were isolated from the aerial part of the mangrove plant Ceriops tagal. The structures and relative configurations were elucidated on the basis of their spectroscopic data. Cytotoxicity of the isolated compounds against HeLa human cervical carcinoma cancer cell line was evaluated.


Assuntos
Diterpenos/isolamento & purificação , Rhizophoraceae/química , Antineoplásicos Fitogênicos/análise , Células HeLa , Humanos , Estrutura Molecular
13.
J Agric Food Chem ; 57(15): 6649-55, 2009 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-19588925

RESUMO

An HPLC method was developed and validated to compare the chemical profiles and tyrosinase inhibitors in the woods, twigs, roots, and leaves of Artocarpus heterophyllus . Five active tyrosinase inhibitors including dihydromorin, steppogenin, norartocarpetin, artocarpanone, and artocarpesin were used as marker compounds in this HPLC method. It was discovered that the chemical profiles of A. heterophyllus twigs and woods are quite different. Systematic chromatographic methods were further applied to purify the chemicals in the twigs of A. heterophyllus. Four new phenolic compounds, including one isoprenylated 2-arylbenzofuran derivative, artoheterophyllin A (1), and three isoprenylated flavonoids, artoheterophyllin B (2), artoheterophyllin C (3), and artoheterophyllin D (4), together with 16 known compounds, were isolated from the ethanol extract of the twigs of A. heterophyllus. The structures of compounds 1-4 were elucidated by spectroscopic analysis. However, the four new compounds did not show significant inhibitory activities against mushroom tyrosinase compared to kojic acid. It was found that similar compounds, such as norartocarpetin and artocarpesin in the twigs and woods of A. heterophyllus, contributed to their tyrosinase inhibitory activity.


Assuntos
Artocarpus/química , Monofenol Mono-Oxigenase/antagonistas & inibidores , Peptídeos/química , Extratos Vegetais/química , Agaricales/enzimologia , Proteínas Fúngicas/antagonistas & inibidores , Casca de Planta/química , Raízes de Plantas/química
14.
J Nat Prod ; 71(4): 674-7, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18303852

RESUMO

Five new lindenane sesquiterpene dimers ( 1- 5), named shizukaols K-O, and eight known sesquiterpene dimers were isolated from the roots of Chloranthus fortunei. The structures of 1- 5 were elucidated using spectroscopic data, mainly 1D NMR, 2D NMR, and mass spectra.


Assuntos
Medicamentos de Ervas Chinesas/isolamento & purificação , Magnoliopsida/química , Sesquiterpenos/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Raízes de Plantas/química , Sesquiterpenos/química
15.
Zhongguo zhenjiu ; (12): 169-170, 2005.
Artigo em Chinês | WPRIM | ID: wpr-245172

RESUMO

<p><b>OBJECTIVE</b>To search for an effective method for treatment of obstinate hiccup.</p><p><b>METHODS</b>Ninety cases of obstinate hiccup secondary to cerebral diseases were randomly divided into 3 groups, 30 cases in each group. Group A were treated by injection of compound chlorpromazine into Yintang (Ex-HN 3), group B by intramuscular injection of compound chlorpromazine and group C by acupuncture at Yintang (Ex-HN 3). Their therapeutic effects were compared.</p><p><b>RESULTS</b>The effective rate was 93.3% in the group A, 10.0% in the group B and 30.0% in the group C, the group A being better than the group B and the group C (P < 0.01).</p><p><b>CONCLUSION</b>Injection of compound chlorpromazine into Yintang (Ex-HN 3) has a significant therapeutic effect on obstinate hiccup.</p>


Assuntos
Humanos , Pontos de Acupuntura , Terapia por Acupuntura , Encefalopatias , Soluço , Injeções
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