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Owing to its ability to separate substances with a broad scope of polarities, exploring the three-phase solvent systems (TPSSs) with high-speed countercurrent chromatography is a topic of interest in separation science, and their retention volumes should be more concerned. This study primarily investigates the behavior of retention volumes while examining the isolation abilities of the TPSS in the technique above. We took standard compounds, including sophoricoside, Sudan red 7B, and rotenone, which have a broad range of polarity, for investigation in this study and separated them using different four-liquid TPSSs made up of water, acetonitrile, methyl acetate, and n-hexane (WAMH). Our findings show that the retention volumes gradually alter in response to changes in phase polarity within the proposed solvent systems. With TPSSs, we preliminarily studied compound isolation and the promising formula of their retention volumes. The proposed solvent systems WAMH in different ratios showed high correlations and adjusted correlation coefficients above 0.9978 and 0.9913 for the actual and calculated retention volumes. This study will be particularly beneficial for researchers focusing on countercurrent chromatography with TPSSs, as it offers valuable time-saving insights.
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Distribuição Contracorrente , Solventes , Solventes/química , Hexanos/química , Acetonitrilas/química , Compostos Azo/química , Água/químicaRESUMO
Introduction: Women with overweight or obesity and gestational diabetes mellitus (GDM) are at a high risk of developing type 2 diabetes mellitus (T2DM) and other metabolic diseases. Healthy postpartum lifestyles in women with GDM are important for effectively preventing early T2DM occurrence; however, few studies and guidelines focus in China on this issue. Aims: This qualitative study aimed to understand the puerperium experience and lifestyle of women with overweight/obesity and GDM. Methods: A face-to-face, in-depth, and semi-structured interview was conducted using a hermeneutical phenomenology method to collect data that were analyzed through thematic analysis. Results: Out of 61 recruited women with overweight/obesity and history of GDM, 14 women underwent an interview and provided detailed descriptions of their lifestyle experiences during puerperium. The interview data were used to generate four themes-puerperium dietary behavior, weight perception and "confinement" behavior, family support, disease knowledge, and perceived risk-and nine sub-themes. Conclusion: Unhealthy lifestyles, misconceptions about food, the conflict between physical activity and confinement behavior, a lack of social and family support, and low awareness of disease risk are all common among overweight/obese women with a history of GDM. Thus, we emphasized that healthcare providers should provide continuous preventive care from pregnancy to postpartum and promote long-term health in high-risk populations with a history of GDM associated with overweight/obesity.
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OBJECTIVE@#To observe the effects of Guizhi Fuling Capsule (GZFLC) on myeloma cells and explore the mechanisms.@*METHODS@#MM1S and RPMI 8226 cells were co-cultured with different concentrations of serum and the cell experiments were divided into negative (10%, 20% and 40%) groups, GZFLC (10%, 20%, and 40%) groups and a control group. Cell counting kit-8 (CCK-8) assays and flow cytometry were used to detect the viability and apoptosis levels of myeloma cells. The effects on mitochondria were examined by reactive oxygen specie (ROS) and tetrechloro-tetraethylbenzimidazol carbocyanine iodide (JC-1) assays. Western blot was used to detect the expression of B cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), cleaved caspase-3, -9, cytochrome C (Cytc) and apoptotic protease-activating factor 1 (Apaf-1). RPMI 8226 cells (2 × 107) were subcutaneously inoculated into 48 nude mice to study the in vivo antitumor effects of GZFLC. The mice were randomly divided into four groups using a completely randomized design, the high-, medium-, or low-dose GZFLC (840, 420, or 210 mg/kg per day, respectively) or an equal volume of distilled water, administered daily for 15 days. The tumor volume changes in and survival times of the mice in the GZFLC-administered groups and a control group were observed. Cytc and Apaf-1 expression levels were detected by immunohistochemistry.@*RESULTS@#GZFLC drug serum decreased the viability and increased the apoptosis of myeloam cells (P<0.05). In addition, this drug increased the ROS levels and decreased the mitochondrial membrane potential (P<0.01). Western blot showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups, whereas the expression levels of cleaved caspase-3, -9, Cytc and Apaf-1 were increased (all P<0.01). Over time, the myeloma tumor volumes of the mice in the GZFLC-administered groups decreased, and survival time of the mice in the GZFLC-administered groups were longer than that of the mice in the control group. Immunohistochemical analysis of tumor tissues from the mice in the GZFLC-administered groups revealed that the Cytc and Apaf-1 expression levels were increased (P<0.05).@*CONCLUSION@#GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway and significantly reduced the tumor volumes in mice with myeloma, which prolonged the survival times of the mice.
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Camundongos , Animais , Caspase 3/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Wolfiporia , Mieloma Múltiplo/tratamento farmacológico , Proteína X Associada a bcl-2/metabolismo , Camundongos Nus , Apoptose , Mitocôndrias/metabolismoRESUMO
Public exposure to radon has attracted increasing public concern. The newly issued "Standards for indoor air quality (GB/T 18883-2022)" has revised the radiological parameters of radon. This study analyzed and discussed the relevant technical contents about the derivation of radon limit, including the distribution level for indoor radon, exposure pathway, health effects, and the process for establishing the standard limits. Specific implementation and evaluation suggestions are also proposed.
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Humanos , Radônio/análise , Poluição do Ar em Ambientes Fechados , China , HabitaçãoRESUMO
Public exposure to radon has attracted increasing public concern. The newly issued "Standards for indoor air quality (GB/T 18883-2022)" has revised the radiological parameters of radon. This study analyzed and discussed the relevant technical contents about the derivation of radon limit, including the distribution level for indoor radon, exposure pathway, health effects, and the process for establishing the standard limits. Specific implementation and evaluation suggestions are also proposed.
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Humanos , Radônio/análise , Poluição do Ar em Ambientes Fechados , China , HabitaçãoRESUMO
OBJECTIVE To pr ovide theoretic support for Guiyang to scientifically guide the development of drug retail industry and implement national health policies . METHODS The data were collected through statistical yearbook ,data cloud , coordinate acquisition device of Application Programming Interface of Baidu map and so on. The spatial distribution characteristics and accessibility of medical insurance designated retail pharmacies (shorted for “designated pharmacies ”)in Guiyang were analyzed by spatial analysis based on Geographic Information System. The related factors for the distribution of designated pharmacies in Guiyang were analyzed by statistical method. RESULTS The number of designated pharmacies ,designated pharmacies per thousand people and designated pharmacies per 10 km2 in Guiyang increased from 2 018,0.41 and 2.51 in 2020 to 2 500,0.42 and 3.11 in 2021,with growth rates of 23.89%,2.44% and 23.90% respectively. The service area of the designated pharmacies that residents of Guiyang reached within 15 minutes on foot was 10.27% of the total service area of designated pharmacies in Guiyang. The results of correlation analysis showed that the correlation coefficients between the regional gross regional production ,total retail sales of consumer goods ,population,urban per capita disposable income and the number of designated pharmacies in Guiyang were 0.999,0.999,0.977 and 0.992,respectively (all P<0.05). CONCLUSIONS The distribution of designated pharmacies is insufficient in Guiyang ,the development of designated pharmacies in various administrative regions is uneven ,and the layout of pharmacies is significantly affected by economic and demographic factors. It is suggested that the local government should explore the strategy of scientifically and reasonably expanding the coverage of designated pharmacies in urban and rural areas,promote the rational layout of pharmacies with appropriate economic and demographic policies ,and pay attention to improving the service capacity of designated pharmacies ,so as to improve the quality of life of the people and guide the healthy and high-quality development of drug retail industry.
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Ten new (1-10) and 26 known (11-36) compounds were isolated from Penicillium griseofulvum MCCC 3A00225, a deep sea-derived fungus. The structures of the new compounds were determined by detailed analysis of the NMR and HRESIMS spectroscopic data. The absolute configurations were established by X-ray crystallography, Marfey's method, and the ICD method. All isolates were tested for in vitro anti-food allergic bioactivities in immunoglobulin (Ig) E-mediated rat basophilic leukemia (RBL)-2H3 cells. Compound 13 significantly decreased the degranulation release with an IC50 value of 60.3 µM, compared to that of 91.6 µM of the positive control, loratadine.
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Antialérgicos/farmacologia , Basófilos/efeitos dos fármacos , Degranulação Celular/efeitos dos fármacos , Hipersensibilidade Alimentar/tratamento farmacológico , Penicillium/metabolismo , Animais , Antialérgicos/isolamento & purificação , Basófilos/imunologia , Linhagem Celular Tumoral , Hipersensibilidade Alimentar/imunologia , Sedimentos Geológicos/microbiologia , Imunoglobulina E/imunologia , Estrutura Molecular , Ratos , Relação Estrutura-AtividadeRESUMO
Acute intestinal graft-versus-host disease is a refractory disease which can affect implantation and become a threat to life in severe cases. Autophagy is an intracellular degradation pathway necessary for maintaining cellular energy homeostasis. In recent years, a large number of studies have found that it is closely related to the pathogenesis and process of acute intestinal graft-versus-host disease. The main mechanisms may involve that inflammatory factor storm after pretreatment and infusion of donor cells induces disordered intestinal immune tolerance, and abnormal oxidative stress damages intestinal mucosal barrier, leading to intestinal rejection of acute graft-versus-host disease via mTOR signal pathway of autophagy, disordered mitophagy and other related pathways.
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Humanos , Autofagia , Doença Enxerto-Hospedeiro , Tolerância Imunológica , Estresse Oxidativo , Transdução de SinaisRESUMO
OBJECTIVE:To establish the method for determining protein binding rate of dipeptidyl peptidase- 4 inhibitor LGT- 6 in different species of plasma ,and to compare their difference. METHODS :By equilibrium dialysis ,LGT-6(3,30,300,3 000 nmol/L)was equilibrated in rat ,monkey and human plasma (i. e. internal dialysis solution )for 48 h,using phosphate buffer as the external dialysis solution. The concentration of LGT- 6 in internal and external dialysis solution was determined by UPLC-MS/MS using tolbutamide as internal standard ,and the plasma protein binding rate was calculated. The determination was performed on ACQUITY UPLC HSS T 3 column with water (containing 0.01% formic acid )-acetonitrile(containing 0.01% formic acid )as mobile phase at the flow rate of 0.6 mL/min. The column temperature was 40 ℃,and the sample size was 2 μL. The ion source was electrospray ion source ,and the multiple ion monitoring mode was used to carry out positive ionization scanning. The ion pairs for quantitative analysis were m/z 487.0→434.3(LGT-6),m/z 271.1→172.0(internal standard ),respectively. RESULTS :At the concentrations of 3,30,300,and 3 000 nmol/L,the protein binding rates of LGT- 6 in rat plasma were (96.25±0.97)%,(84.16± 1.24)%,(78.25±0.61)%,(66.63±0.95)%;the protein protein binding rates in monkey plasma were (98.54±0.58)%,(87.27± 1.01)%,(79.35±0.86)%,(66.69±0.54)%;the protein binding rates in human plasma were (99.40±1.03)%,(84.48± 1.15)%,(77.62±0.77)%,(66.93±0.48)%. At the same concentration ,the protein binding rates of LGT- 6 in rat ,monkey and human plasma had no significant difference (P>0.05). In the same species of plasma ,there were significant differences in the plasma protein binding rates of different concentration of LGT-6 among those groups (P<0.05),and it decreased with 才〔2016〕4015) the increase of drug concentration. CONCLUSIONS : The method for the determination of plasma protein binding rate of LGT-6 is successfully established. The data revealed that the protein binding rate of LGT- 6 is concentration-dependent , there was no obvious spec ies difference on protein binding rates of LGT- 6 in rat ,monkey and human plasma under the same concentration.
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OBJECTIVE:To establish the metho d for the concentration det ermination of foretinib derivative LWK- 126 in liver microsomes,and to study its metabolism stability in liver microsomes of rats ,Beagle dogs and human. METHODS :In the in vitro incubation system of liver microsomes ,LWK-126 was dissolved in liver microsomal incubation systems of rats ,Beagle dog and human initiated by reduced nicotinamide adenine dinucleotide phosphate solution. After incubation in water at 37 ℃ for 0,5,10,20, 30 and 60 min,the reaction was terminated with acetonitrile containing internal standard (1 μg/mL tolbutamide). UPLC-MS/MS method was applied to determine the concentration of LWK- 126 in the incubation systems. The determination was performed on Waters BEH C 18 column with mobile phase consisted of water (containing 0.1% formic acid )-acetonitrile(containing 0.1% formic acid)by gradient elution at the flow rate of 0.4 mL/min. The column temperature was 30 ℃,and the sample size was 2 μL. The mass spectral analysis was performed in a positive electrospray ionization mode ,and the full MS experiment was run with the selective reaction monitoring mode with a scanning range of m/z 50→1 200. Taking the concentration of LWK- 126 at 0 min as reference,the remaining percentage and the enzyme kinetic parameters were calculated. RESULTS :The linear range of LWK- 126 was 0.05-15 μg/mL(R 2=0.999 2);the lower limit of quantification was 0.05 μg/mL,and the lowest detection limit was 0.01 μg/mL. The precision,accuracy,extraction recovery and matrix effect all met the analysis requirements of biological samples. The remaining percentage of LWK- 126 in liver microsomes of human ,rats and Beagle dogs for 60 min were (33.17±4.52)%,(3.14± 6.73)%,(1.38±5.85)%;t1/2 of them were 33.15,11.76,5.62 min;the clearance rates were 38.45,118.81,245.76 μL(/ min·mg), respectively. CONCLUSIONS :The method for the content ; determination of LWK- 126 in liver microsomes is established successfully. The order of metabolic stability of LWK- 126 in 〔2016〕4015) liver microsomes of different species is human >rats>Beagle dogs.
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From the deep-sea-derived fungus Aspergillus candidus, one novel (1) and three known (2-4) p-terphenyl derivates were isolated. The structure of the new compound was established mainly on the basis of extensive analysis of 1D and 2D NMR data. All four isolates were tested for in vitro anti-food allergic and antitumor bioactivities. Compounds 3 and 4 showed potent antiproliferative effect against four cancer cells of Hela, Eca-109, Bel-7402, and PANC-1 with IC50 values ranging from 5.5 µM to 9.4 µM.
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Aspergillus/química , Oceanos e Mares , Compostos de Terfenil/farmacologia , Antineoplásicos/farmacologia , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Espectroscopia de Prótons por Ressonância Magnética , Compostos de Terfenil/química , Compostos de Terfenil/isolamento & purificaçãoRESUMO
Some non-coding RNAs (ncRNA), as functional RNA molecules, lack potential to encode proteins, but can affect gene expression and disease progression through a variety of mechanisms. In multiple myeloma (MM), cardiovascular disease is one of the most common complications, which may be related to a variety of factors, including patient's own factors, disease-related factors, drug factors, etc. Non-coding RNA is considered to be an important regulator of cardiovascular event risk factors and cell function, and an important candidate target for improving the condition and prognostic assessment. This article briefly summarized the role of non-coding RNA in cardiac amyloidosis caused by MM, damage to the heart by inflammatory factors, and heart disease caused by chemotherapy drugs in recent years.
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Humanos , Doenças Cardiovasculares , Cardiopatias , Mieloma Múltiplo/genética , Prognóstico , RNA não Traduzido/genéticaRESUMO
Four new (penigrisacids A-D, 1-4) and one known (5) carotane sesquiterpenoids were isolated from the deep-sea-derived fungus Penicillium griseofulvum, along with four known compounds (6-9). The planar structures and relative configurations of the new compounds were determined by extensive analysis of the NMR and HRESIMS data. The absolute configurations were established by comparison of the experimental and calculated ECD (electronic circular dichroism) spectra or OR (optical rotation) value. Compound 9 exhibited potent anti-food allergic activity with IC50 value of 28.7 µM, while 4 showed weak cytotoxicity against ECA-109 tumor cells (IC50 = 28.7 µM).
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Organismos Aquáticos/química , Penicillium/química , Sesquiterpenos/química , Linhagem Celular Tumoral , Dicroísmo Circular , Células HeLa , Humanos , Espectroscopia de Ressonância Magnética/métodos , Oceanos e Mares , Sesquiterpenos/farmacologiaRESUMO
A novel ergostane, sarocladione (1), was isolated from the deep-sea-derived fungus Sarocladium kiliense, along with 20 known compounds. The structure of 1 was determined mainly by a detailed analysis of its experimental and calculated NMR spectroscopic data. It is worth noting that 1 was the first steroid bearing a 5,10:8,9-diseco moiety. All 21 compounds were tested for in vitro antitumor activities against five cancer cell lines. ß-Sitostenone (7) and 4,6-dihydroxyeudesmane (20) showed significant effects on HeLa-S3 cells with the IC50 values of 9.2 µM and 9.3 µM, respectively.
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Acremonium/química , Antineoplásicos/farmacologia , Secoesteroides/farmacologia , Esteroides/farmacologia , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Células HeLa , Humanos , Conformação Molecular , Secoesteroides/química , Secoesteroides/isolamento & purificação , Esteroides/química , Esteroides/isolamento & purificação , Relação Estrutura-AtividadeRESUMO
Background: Inflammation and altered immunity contribute to the development of pulmonary arterial hypertension (PH). The alpha 7 nicotinic acetylcholine receptor (α7nAChR) possesses anti-inflammatory activities. The current study was performed to investigate the effects of a selective α7nAChR agonist, PNU-282987, on controlling a monocrotaline (MCT)-induced rat model of PH and explored the underlying mechanisms. Methods: Sprague-Dawley rats were injected with MCT and treated with PNU-282987 at the prevention (starting 1 week before MCT) and treatment (starting 2 weeks after MCT) settings. Four weeks after MCT injection, hemodynamic changes, right ventricular structure, and lung morphological features were assessed. Enzyme-linked immunosorbent assay, Western blot and qRT-PCR were performed to assess levels of inflammatory cytokines and NLRP3 (Nod-like receptor family pyrin domain-containing 3) inflammasome pathway in the rat lung tissues. In addition, the lung macrophage line NR8383 was used to confirm the in vivo data. Results: Monocrotaline injection produced PH in rats and downregulated α7nAChR mRNA and protein expression in rat lung tissues compared to sham controls. Pharmacological activation of α7nAChR by PNU-282987 therapy improved the rat survival rate, attenuated the development of PH as assessed by remodeling of pulmonary arterioles, reduced the right ventricular (RV) systolic pressure, and ameliorated the hypertrophy and fibrosis of the RV in rats with MCT-induced PH. The expression of TNF-α, IL-6, IL-1ß, and IL-18 were downregulated in rat lung tissues, which implied that PNU-282987 therapy may help regulate inflammation. These protective effects involved the inhibition of the NLRP3 inflammasome. In vitro assays of cultured rat lung macrophages confirmed that the anti-inflammation effect of PNU-282987 therapy may contribute to the disturbance of NLRP3 inflammasome activation. Conclusion: Targeting α7nAChR with PNU-282987 could effectively prevent and treat PH with benefits for preventing ongoing inflammation in the lungs of rats with MCT-induced PH by inhibiting NLRP3 inflammasome activation.
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BACKGROUND: Neurological diseases have become an obvious challenge due to insufficient therapeutic intervention. Therefore, novel drugs for various neurological disorders are in desperate need. Recently, compelling evidence has demonstrated that chemokine receptor CXCR3, which is a G protein-coupled receptor in the CXC chemokine receptor family, may play a pivotal role in the development of neurological diseases. The aim of this review is to provide evidence for the potential of CXCR3 as a therapeutic target for neurological diseases. METHODS: English journal articles that focused on the invovlement of CXCR3 in neurological diseases were searched via PubMed up to May 2017. Moreover, reference lists from identified articles were included for overviews. RESULTS: The expression level of CXCR3 in T cells was significantly elevated in several neurological diseases, including multiple sclerosis (MS), glioma, Alzheimer's disease (AD), chronic pain, human T-lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and bipolar disorder. CXCR3 antagonists showed therapeutic effects in these neurological diseases. CONCLUSION: These studies provided hard evidence that CXCR3 plays a vital role in the pathogenesis of MS, glioma, AD, chronic pain, HAM/TSP and bipolar disorder. CXCR3 is a crucial molecule in neuroinflammatory and neurodegenerative diseases. It regulates the activation of infiltrating cells and resident immune cells. However, the exact functions of CXCR3 in neurological diseases are inconclusive. Thus, it is important to understand the topic of chemokines and the scope of their activity in neurological diseases.
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Doenças do Sistema Nervoso/metabolismo , Receptores CXCR3/metabolismo , Animais , Movimento Celular , Sobrevivência Celular , Modelos Animais de Doenças , Humanos , Doenças do Sistema Nervoso/etiologia , Doenças do Sistema Nervoso/imunologia , Linfócitos T/metabolismoRESUMO
Flaviviridae family is a class of single-stranded RNA virus, which is fatal to human and animals and mainly prevalent in subtropic and tropic countries. Even though people and animals are barraged with flavivirus infection every year, we have not invented either vaccines or antiviral for most flavivirus infections yet. Innate immunity is the first line of defense in resisting pathogen invasion, serving an important role in a resisting virus. Toll-like receptors (TLRs) and retinoic acid-inducible gene I- (RIG-I-) like receptors (RLRs) are crucial pattern recognition receptors (PRRs) that play essential roles in recognizing and clearing pathogens, including resisting flavivirus. In the present review, we provide a significant reference for further research on the function of innate immunity in resisting flavivirus.
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Proteína DEAD-box 58/metabolismo , Resistência à Doença/imunologia , Infecções por Flavivirus/imunologia , Infecções por Flavivirus/metabolismo , Flavivirus/imunologia , Interações Hospedeiro-Patógeno/imunologia , Receptores Toll-Like/metabolismo , Animais , Infecções por Flavivirus/virologia , Humanos , Transdução de SinaisRESUMO
Objective To investigate the clinical efficacy of acupuncture combined with back Shu acupoint catgut embedding therapy in treating allergic rhinitis. Methods Totally 66 allergic rhinitis patients were included. Random number table method was used to divide the 66 patients into observation group and control group, with 33 cases in each group. The control group was given Ketotifen Fumarate Nasal Drops nose drops 2 drops each time, 3 times a day, Loratadine Tablets 10 mg orally, once a day for 4 weeks. The experimental group was treated with acupuncture, once a day, 10 d as a treatment course, three courses in total; at the same time, back Shu acupoint catgut embedding therapy was given, 15 d each time, twice treatment in total. After the end of treatment, clinical efficacy, clinical symptom score, life quality score, serum levels of inflammatory factors, serum immunoglobulin, peripheral blood eosinophil count and the incidence of adverse reactionsof the two groups were observed. Results The total effective rate was 93.94% (31/33) in observation group and 75.76% (25/33) in the control group, with statistical significance in the two groups (P0.05). Conclusion Acupuncture combined with catgut embedding therapy in the treatment of allergic rhinitis has a significant clinical efficacy, which can improve the level of inflammatory factors in patients with high safety.
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<p><b>OBJECTIVE</b>To analyze changes in gene amplification in the mitochondrial genome and in the ID4 gene promoter methylation region in patients with chronic aplastic anemia (CAA) suffering from Kidney (Shen) yin deficiency or Kidney yang deficiency.</p><p><b>METHODS</b>Bone marrow and oral epithelium samples were collected from CAA patients with Kidney yin deficiency or Kidney yang deficiency (20 cases). Bone marrow samples were collected from 20 healthy volunteers. The mitochondrial genome was amplified by polymerase chain reaction (PCR), and PCR products were used for sequencing and analysis.</p><p><b>RESULTS</b>Higher mutational rates were observed in the ND1-2, ND4-6, and CYTB genes in CAA patients suffering from Kidney yin deficiency. Moreover, the ID4 gene was unmethylated in bone marrow samples from healthy individuals, but was methylated in some CAA patients suffering from Kidney yin deficiency (positive rate, 60%) and Kidney yang deficiency (positive rate, 55%).</p><p><b>CONCLUSIONS</b>These data supported that gene mutations can alter the expression of respiratory chain enzyme complexes in CAA patients, resulting in energy metabolism impairment and promoting the physiological and pathological processes of hematopoietic failure. Functional impairment of the mitochondrial respiration chain induced by gene mutation may be an important reason for hematopoietic failure in patients with CAA. This change is closely related to maternal inheritance and Kidney yin deficiency. Finally, these data supported the assertion that it is easy to treat disease in patients suffering from yang deficiency and difficult to treat disease in patients suffering from yin deficiency.</p>
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Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Anemia Aplástica , Genética , Sequência de Bases , Biópsia , Medula Óssea , Patologia , Estudos de Casos e Controles , Doença Crônica , Metilação de DNA , Genética , DNA Mitocondrial , Genética , Eletroforese em Gel de Ágar , Genoma Mitocondrial , Genética , Proteínas Inibidoras de Diferenciação , Genética , Rim , Patologia , Mutação , Genética , Polimorfismo Genético , Regiões Promotoras Genéticas , Genética , Deficiência da Energia Yin , GenéticaRESUMO
OBJECTIVE: To clone and express the aegyptin-like protein (alALP) encoding gene from Aedes albopictus salivary gland, and analyze its antigenicity. METHODS: The homology, secondary structure and antigen peptides of alALP and aegyptin protein (GenBank No. ABF18122.1) was analyzed by bioinformatics software tools. Total RNA was extracted from Ae. albopictus salivary gland. The coding region of alALP (GenBank No. AY826121) was amplified by PCR. RT-PCR product was digested with restriction enzyme and ligated into a pGEX-6P-1 vector. The recombinant pGEX-6P-1-alALP plasmid was transformed into E. coli BL21 and induced by IPTG. The recombinant soluble GST-alALP fusion protein was purified with Glutathione Sepharose 4B. The expression product was analyzed by SDS-PAGE and Western blotting. Mice were immunized each with 60 microg purified GST-alALP at every 2 weeks for 3 times, and mouse anti-GST-alALP serum was prepared. Western blotting assay with mice anti-GST-alALP serum and serum of mice exposed to Ae. albopictus bites was used to analyze its antigenicity. RESULTS: Bioinformatics prediction results showed that alALP and aegyptin had 65.58% homology with a similar secondary structure, and a conservative polypeptide. The product of RT-PCR was 762 bp. The recombinant plasmid pGEX-6P-1-alALP was confirmed by double restriction enzyme digestion, PCR and sequencing. SDS-PAGE and Western blotting analysis showed that the bacteria containing recombinant plasmid pGEX-6p-1-alALP expressed a soluble recombinant fusion protein (M(r) 56 000) after being induced with IPTG. Western blotting analysis revealed that GST-alALP protein was recognized by mouse anti-GST-alALP serum and serum of mice ex- posed to Ae. albopictus bites. CONCLUSION: Mature peptide gene of alALP can be expressed in prokaryotic expression system, and the recombinant protein shows antigenicity.