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BACKGROUND: Panic disorder (PD) involves emotion dysregulation, but its underlying mechanisms remain poorly understood. Previous research suggests that implicit emotion regulation may play a central role in PD-related emotion dysregulation and symptom maintenance. However, there is a lack of studies exploring the neural mechanisms of implicit emotion regulation in PD using neurophysiological indicators. AIM: To study the neural mechanisms of implicit emotion regulation in PD with event-related potentials (ERP). METHODS: A total of 25 PD patients and 20 healthy controls (HC) underwent clinical eva-luations. The study utilized a case-control design with random sampling, selecting participants for the case group from March to December 2018. Participants performed an affect labeling task, using affect labeling as the experimental condition and gender labeling as the control condition. ERP and behavioral data were recorded to compare the late positive potential (LPP) within and between the groups. RESULTS: Both PD and HC groups showed longer reaction times and decreased accuracy under the affect labeling. In the HC group, late LPP amplitudes exhibited a dynamic pattern of initial increase followed by decrease. Importantly, a significant group × condition interaction effect was observed. Simple effect analysis revealed a reduction in the differences of late LPP amplitudes between the affect labeling and gender labeling conditions in the PD group compared to the HC group. Furthermore, among PD patients under the affect labeling, the late LPP was negatively correlated with disease severity, symptom frequency, and intensity. CONCLUSION: PD patients demonstrate abnormalities in implicit emotion regulation, hampering their ability to mobilize cognitive resources for downregulating negative emotions. The late LPP amplitude in response to affect labeling may serve as a potentially valuable clinical indicator of PD severity.
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Reverse prediction and molecular docking techniques were employed to evaluate the feasibility of reniformin A(RA) as an anti-tumor leading compound. Based on the reverse prediction, network pharmacology was used to construct a "disease-compound-target-pathway" network. Thirty-nine tumor-related targets of RA were predicted, which participated in the regulation of multiple cellular activities such as apoptosis, cell cycle, and tumor metastasis, and regulated estrogen signal transduction and inflammatory response. Discovery Studio 2020 was adopted for molecular docking and toxicity prediction(TOPKAT). As revealed by the results, the binding affinity of RA with the tumor-related targets ABL1, ESR1, SRC and BCL-XL was stronger than that of oridonin(OD), while its mutagenicity, rodent carcinogenesis, and oral LD_(50) in rats were all inferior to that of OD. Furthermore, in vitro experiments were performed to confirm the anti-tumor activity of RA, and the mechanism was preliminarily discussed. The results demonstrated that RA was superior to OD in cytotoxicity, inhibition of cell colony formation, and induction of apoptosis. RA, possessing potent anti-tumor activity, is expected to be a new anti-tumor leading compound.
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Animales , Ratas , Medicamentos Herbarios Chinos/farmacología , Plomo , Simulación del Acoplamiento Molecular , Neoplasias/genética , Transducción de SeñalRESUMEN
Farnesyltransferase (FTase) was selected as a target for virtual screening of inhibitors using the Glide v4.0 program in the Schrödinger software package. We discovered 13 novel structures as farnesyltransferase inhibitors (FTIs) with moderate potency. By analyzing the binding modes of representative compounds 8 (IC50 = 2.29 μmol·L-1) and 18 (IC50 = 0.41 μmol·L-1) with farnesyltransferase, it was found that compounds 8 and 18 didn't coordinate with Zn2+, indicating that the coordination between FTIs with Zn2+ is not essential for the bioactivity of the inhibitors. The structure-activity relationship was summarized by analyzing the predicted binding modes of representative compounds. It was found that the scaffolds of the discovered FTIs had room for structural optimization, which lay foundation for obtaining highly active and selective FTIs.
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The aim of this study is to investigate the protective effects of a small molecular fraction (SMF) of Polygoni multiflori Radix Praeparata (PMRP) in a cyclophosphamide (CTX) induced anemia mouse model. Small molecular fraction of PMRP was prepared and identified by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). In pharmacology, we examined the peripheral hemogram and thymus and spleen index. The content of granulocyte-macrophage colony-stimulating factor (GM-CSF) in serum was mensurated by enzyme-linked immunosorbent assay (ELISA); The level of superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in serum and spleen tissue homogenate were detected, and glutathione peroxidase (GSH-PX) was assayed in spleen. The results show that SMF can significantly accelerate the recovery of peripheral hemogram, increase the activity of antioxidant enzymes and GM-CSF in serum and spleen. SMF also increases the number of spleen cells, improves bone marrow pathology. In conclusion, the SMF of PMRP promoted the recovery of hematopoietic function in a CTX-induced anemia mouse, which can support SMF to be used as an adjunct to chemotherapy to counteract its side effects.
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<p><b>OBJECTIVE</b>To study the relationship between the expression of peripheral blood HLA-DR, CD4CD25 regulatory T cells, IL-17 and IL-27 with liver damage in children with human cytomegalovirus (HCMV) infection.</p><p><b>METHODS</b>Twenty-one HCMV children with liver damage and twenty-one HCMV children without liver damage were enrolled in this study. The expression of peripheral blood HLA-DR and CD4CD25 regulatory T cells was detected by flow cytometry. Plasma levels of IL-17 and IL-27 were measured using ELISA.</p><p><b>RESULTS</b>The plasma levels of IL-17 and IL-27 in children with liver damage were significantly higher than in those without liver damage, while the expression of peripheral blood CD4CD25 regulatory T cells was lower than in those without liver damage (P<0.05). Plasma IL-17 and IL-27 levels were negatively correlated with the expression of peripheral blood CD4CD25 regulatory T cells (P<0.01).</p><p><b>CONCLUSIONS</b>Immune imbalance mediated by CD4CD25 regulatory T cells and over-expression of IL-17 and IL-27 may be involved in the pathogenesis of liver damage in children with HCMV infection.</p>
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Femenino , Humanos , Lactante , Masculino , Antígenos CD4 , Alergia e Inmunología , Citomegalovirus , Fisiología , Infecciones por Citomegalovirus , Sangre , Genética , Citometría de Flujo , Antígenos HLA-DR , Genética , Alergia e Inmunología , Interleucina-17 , Sangre , Genética , Subunidad alfa del Receptor de Interleucina-2 , Alergia e Inmunología , Interleucinas , Sangre , Genética , Hígado , Heridas y Lesiones , Metabolismo , Hepatopatías , Sangre , Alergia e Inmunología , Linfocitos T Reguladores , Alergia e InmunologíaRESUMEN
This study was conducted to improve structural instability of a highly active DHODH inhibitor A found in our group. Twelve prodrugs were synthesized by modifying the carboxyl group. The enzyme activity test of 12 prodrugs A1−A12 demonstrated that A1−A5 displayed weak inhibitory activity, and A6−A12 displayed no activity, which met the action mechanism of designed prodrug. The structural stability of A1−A12 in methanol and pH 2.0, 9.0 buffers were tested, and the results showed that A12 could avoid intramolecular ring-formation in CH3OH, A1−A8 were easily hydrolyzed under acidic conditions, and A9−A12 were inclined to hydrolyze under alkaline conditions. The cell proliferation inhibitory activity of 12 prodrugs were evaluated, in which compound A12 displayed excellent activity (IC50=0.63 μmol·L−1) similar to brequinar. These results laid a good foundation for conducting further vivo studies.
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<p><b>OBJECTIVE</b>To investigate the percentages of peripheral blood γδ T cells and regulatory T cells (Treg) and the expression of associated cytokines, interleukin 17 (IL-17) and transforming growth factor-β1 (TGF-β1), in infants with human cytomegalovirus (HCMV) infection.</p><p><b>METHODS</b>Twenty-two infants with HCMV infection (HCMV group) and 22 healthy infants who underwent physical examination (control group) were enrolled in this study. The percentages of peripheral blood γδ T cells and Treg cells were determined by flow cytometry. The levels of IL-17 and TGF-β1 in plasma were measured using ELISA.</p><p><b>RESULTS</b>Compared with the control group, the HCMV group had significantly higher percentage of γδ T cells and IL-17 level (P<0.01) and significantly lower percentage of Treg cells and TGF-β1 level (P<0.01). In the HCMV group, the percentage of γδ T cells was negatively correlated with the percentage of Treg cells and TGF-β1 level (P<0.05), but positively correlated with IL-17 level (P<0.05); the percentage of Treg cells was positively correlated with TGF-β1 level (P<0.05), but negatively correlated with IL-17 level (P<0.05); there was no correlation between IL-17 level and TGF-β1 level (P>0.05).</p><p><b>CONCLUSIONS</b>There is an imbalance between γδ T cells and Treg cells in the peripheral blood of infants with HCMV infection, and γδ T cells may be involved in the secretion of IL-17.</p>
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Femenino , Humanos , Lactante , Masculino , Citocinas , Sangre , Infecciones por Citomegalovirus , Alergia e Inmunología , Interleucina-17 , Sangre , Receptores de Antígenos de Linfocitos T gamma-delta , Linfocitos T Reguladores , Alergia e Inmunología , Factor de Crecimiento Transformador beta1 , SangreRESUMEN
In this study, 1-(3-(4-chlorophenyl)-5-methylthio-1H-1,2,4-triazol-1-yl)-butan-1-one discovered previously in our lab was selected as a inhibitor of human dihydroorotate dehydrogenase (HsDHODH) for structural optimization. The co-crystal of HsDHODH with the hit was obtained and analyzed for guiding the subsequent structural optimization. As a result, a series of novel triazole derivatives were designed and synthesized as potent HsDHODH inhibitors. Among them, compound (3-(4-chlorophenyl)-5-ethylthio-1H-1,2,4-triazol-1-yl)-furan-2-yl-methanone displayed high potency in the inhibition of HsDHODH with an IC50 value of 1.50 μmol·L-1. Meanwhile, the structure-activity relationships were analyzed based on the biological data and the co-crystal structure. These results provide a valuable reference for optimization of 1H-1,2,4-triazole derivatives as HsDHODH inhibitors in the future.
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In order to effectively assess the impact of the drug zero plus policies on the burden of patients in the process of the separation of clinic from pharmacy in Shanghai,the cost data of a drug zero plus hospital and the maternal hospital from 2013 to 2014 was collected,and then the average cost,average pharmacy cost,average person cost,return rate and the ratio of pharmacy were analyzed.The results show the indicators of the burden of patients decreased significantly,while it was suggested that the government needs to increase compensation,continue to adjust the medical price and strengthen medical insurance to persistently promote the separation of clinic from pharmacy.
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Objective: To discuss the acupoints selection pattern in acupuncture-moxibustion treatment of perimenopausal syndrome (PMS) from 2007 to 2016. Methods: Clinical literatures related to PMS treated with acupuncture-moxibustion published from 2007 to 2016 were collected from Chinese Biomedical Literature Database (CBM), Chongqing VIP Database (CQVIP), China National Knowledge Infrastructure (CNKI), and Wanfang Academic Journal Full-text Database (Wanfang). The retrieved data underwent descriptive analysis, cluster analysis and association pattern analysis to determine the acupoints selection principle in acupuncture-moxibustion treatment of PMS. Results: The top five acupoints used in acupuncture-moxibustion treatment of PMS were Sanyinjiao (SP 6), Shenshu (BL 23), Guanyuan (CV 4), Baihui (GV 20), and Shenmen (HT 7). The leading 4 meridians were Bladder Meridian, Conception Vessel, Spleen Meridian, and Governor Vessel. The clustering analysis showed that the 5 core acupoint groups were: ① Sanyinjiao (SP 6); ② Shenshu (BL 23) and Guanyuan (CV 4); ③ Baihui (GV 20), Shenmen (HT 7), Zusanli (ST 36), Ganshu (BL 18) and Taichong (LR 3); ④ Taixi (KI 3), Pishu (BL 20), Xinshu (BL 15), Qihai (CV 6) and Neiguan (PC 6); ⑤ Sishencong (EX-HN 1), Zhongwan (CV 12), Hegu (LI 4), Yintang (GV 29), Fengchi (GB 20), Zhongji (CV 3) and Feishu (BL 13). The three most significant acupoints were Sanyinjiao (SP 6), Shenshu (BL 23) and Guanyuan (CV 4). Acupoint groups based on syndrome differentiation included: ① Hegu (LI 4), Zhongwan (CV 12) and Sishencong (EX-HN 1); ② Feishu (BL 13), Zhongji (CV 3), Fengchi (GB 20) and Yintang (GV 29); ③ Xinshu (BL 15), Pishu (BL 20), Qihai (CV 6), Neiguan (PC 6) and Taixi (KI 3);④ Ganshu (BL 18), Zusanli (ST 36), Shenmen (HT 7), Taichong (LR 3) and Baihui (GV 20). The analysis of association pattern elaborated that Shenshu (BL 23) and Sanyinjiao (SP 6) won the highest support rate in the paired groups; Ganshu (BL 18), Shenshu (BL 23) and Sanyinjiao (SP 6) had the highest support rate among the acupoint groups. Conclusion: The data mining results of acupuncture-moxibustion treatment of PMS substantially conform to the general principle in traditional acupuncture-moxibustion theories, able to reflect the acupoints selection and grouping pattern and provide references for acupuncture-moxibustion treatment of PMS.
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<p><b>OBJECTIVE</b>To evaluate the effect of capsaicin on nude mice xenografted with colorectal carcinoma cells, and to explore its mechanism of action.</p><p><b>METHODS</b>A nude mouse model of colorectal cancer was established by subcutaneous inoculation of human colorectal carcinoma HT-29 cells. Terminal deoxynucleotidyl transferase-mediated nicked labeling assay (TUNEL) was undertaken to detect the cell proliferation and apoptosis in the xenograft tissue in nude mice. Immunohistochemical (IHC) staining and Western blot were used to detect the expression of HSP27, Cyt-C and active caspase-3.</p><p><b>RESULTS</b>The tumor growth of the groups C10 and C20 was significantly slower than that of the group NS. The integrated optical density (IOD) of both the group C5 (2532.14 ± 578.11) and group C10 (6364.03 ± 1137.98) was significantly higher than that of the group NS (760.12 ± 238.05), (P < 0.05). The integrated optical density (IOD) of the group C20 was (15743.96 ± 1855.95), significantly higher than that of the groups C10, C5 and NS (all were P < 0.01). Immunohistochemistry showed that the cytoplasmic expression of HSP27 was strongly positive in the group NS, and significantly reduced with the increasing dose of capsaicin in the treated groups. The expression of active caspase-3 and Cyt-C in the group NS was weakly positive, and was significantly increased with the increasing dose of capsaicin in the groups C5 and C10 (P < 0.05), and the expression of active caspase-3 and Cyt-C of the group C20 was significantly higher than that of the groups C5, C10 and NS (P < 0.01). Western blot analysis showed that both the expressions of HSP27 of the group C5 (0.73 ± 0.05) and the group C10 (0.41 ± 0.03) were significantly lower than that of the group NS (P < 0.05). The expression of HSP27 of the group C20 (0.22 ± 0.06) was significantly lower than that of the groups C5, C10 and NS (P < 0.01). The expressions of active-caspase-3 and Cyt-C in the group C5 were (2.57 ± 0.34) and (2.03 ± 0.38), significantly higher than those of the group NS (P < 0.05). The expressions of active-caspase-3 and Cyt-C in the group C10 were (4.23 ± 0.45) and (3.13 ± 0.44), also significantly higher than those of the group NS (P < 0.05). The expressions of active-caspase-3 and Cyt-C in the group C20 were (5.78 ± 0.48) and (4.92 ± 0.52), significantly higher than those of the group C5, C10 and NS (P < 0.01). TUNEL analysis showed that there was a significant difference of cell apoptosis in comparison of each two groups. The higher dose of capsaicin was used, the more apoptosis was observed.</p><p><b>CONCLUSIONS</b>Capsaicin can significantly inhibit the tumor growth and induce cell apoptosis in the colorectal carcinoma xenograft in nude mice. Its mechanism of action is possibly related with the down-regulation of HSP27 expression and up-regulation of expression of active caspase-3 and Cyt-C in the colorectal carcinoma xenograft in nude mice.</p>
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Animales , Femenino , Humanos , Ratones , Antineoplásicos Fitogénicos , Farmacología , Apoptosis , Capsaicina , Farmacología , Caspasa 3 , Metabolismo , Proliferación Celular , Grupo Citocromo c , Metabolismo , Relación Dosis-Respuesta a Droga , Proteínas de Choque Térmico HSP27 , Metabolismo , Células HT29 , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Distribución Aleatoria , Carga Tumoral , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
<p><b>OBJECTIVE</b>To observe the effects of curing the cervical spondylotic radiculopathy with the methods of Blade needle closed loosing and reduction with traction at the same time.</p><p><b>METHODS</b>From May 2006 to May 2009, 65 patients with cervical spondylotic radiculopathy were divided into treatment group and control group according the random number table produced by SAS Software. There were 18 males and 17 females in the treatment group,age in range from 42 to 73 years old with an average of (61.3 +/- 6.4) years, course of disease was from 8 to 42 months with an average of (23.8 +/- 13.8) months, preoperatively cervical functional score was from 4 to 17 scores with the mean of (11.45 +/- 3.31) scores. And in the control group, including 14 males and 16 females, aged from 44 to 76 years old with an average of (62.4 +/- 8.8) years, course of disease was from 7 to 43 months with an average of (24.4 +/- 16.8) months, preoperatively cervical functional score was from 4 to 18 scores with the mean of (11.40 +/- 3.24) scores. The patients of treatment group were treated with Blade needle closed loosing the specific pain point on the neck and shoulder, then immediately underwent traction and reduction after operation. And the patients of control group were treated with traditional traction. The cervical functional score were compared between the two groups at 10, 20, 180 d after treatment, including pain of neck and shoulder, limitation of motion of neck, tenderness of neck, numbness and muscle weakness of upper limb.</p><p><b>RESULTS</b>(1) At the 10th day after treatment, the total score of treatment group was (15.43 +/- 3.46) scores, which was obviously higher than that of control group's (13.17 +/- 3.18) scores (P < 0.01). In different symptoms, treatment group also was better than that of control group (P < 0.05), so as in the tenderness of neck, and especially in the limitation of motion of neck and muscle weakness of upper limb decreased obviously (P < 0.01). (2) At the 20th day after treatment, the total score of treatment group was (18.00 +/- 2.94) scores, which was obviously better than that of control group's (15.90 +/- 2.89) scores (P < 0.01). In different symptoms, treatment group also was better than that of control group (P < 0.05), so as in the pain of neck and shoulder, numbness and muscle weakness of upper limb (P < 0.05), and especially in the limitation of motion of neck, tenderness of neck decreased obviously (P < 0.01). (3) At the 180th day after treatment, the total score of treatment group was (16.63 +/- 3.32) scores, which was obviously better than that of control group's (12.67 +/- 3.42) scores (P < 0.01); In different symptoms, treatment group also was better than that of control group (P < 0.05), so as in the numbness of upper limb (P < 0.05), and especially in the pain of neck and shoulder, muscle weakness of upper limb, limitation of motion of neck, tenderness of neck decreased obviously (P < 0.01).</p><p><b>CONCLUSION</b>Compared with method of traditional traction, Blade needle closed loosing and traction in treating cervical spondylotic radiculopathy can significantly obtain clinical effects,which can quickly improve symptoms, relieve pain of neck and shoulder, limitation of motion of neck, tenderness of neck, numbness and muscle weakness of upper limb.</p>
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Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios de Casos y Controles , Terapia Combinada , Medicina Tradicional China , Radiculopatía , Terapéutica , Espondilosis , Terapéutica , Tracción , MétodosRESUMEN
<p><b>OBJECTIVE</b>To investigate the possible mechanism of local hyperthermia in the treatment of warts through detecting the differences in CD1a/CD83 of Langerhans cells (LCs) in émigrés from HPV-infected skin, as compared to normal skin.</p><p><b>METHODS</b>Confocal microscopy were performed on Condyloma Accuminatum (CA)and normal skin; Freshly taken biopsies of CA and normal skin were subjected to surface heating at 37 degrees C, 42 degrees C and 45 degrees C respectively, for 30 mins. Flow cytometry was used to determine the CD1a/ CD83 changes of LCs in émigrés from CA and normal skin.</p><p><b>RESULTS</b>By confocal microscopic observation, there were practically no CD1a+ LCs that expressed CD83 in the epidermis of both normal skin and CA. The proportions of CD1a+/CD83 LCs were significantly increased with increased temperatures in émigrés from both normal skin and CA. At each given temperature, the numbers of LCs in émigrés from CA were greater than those from normal skin.</p><p><b>CONCLUSION</b>Local hyperthermia can promote migration and maturation of LCs in HPV-infected skin and accordingly stimulate the immune system to treat warts.</p>
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Adulto , Femenino , Humanos , Adulto Joven , Movimiento Celular , Alergia e Inmunología , Hipertermia Inducida , Técnicas In Vitro , Células de Langerhans , Biología Celular , Alergia e Inmunología , Papillomaviridae , Alergia e Inmunología , Virulencia , Piel , Alergia e Inmunología , VirologíaRESUMEN
OBJECTIVE@#To explore the association of metabolic syndrome(MS) with serum interleukin-10 (IL-10) and high sensitive C reactive protein(hs-CRP) in old men with MS.@*METHODS@#Seventy patients with MS and 30 age-matched controls were enrolled. Blood pressure, waist circumference(WC), weight, height, body mass index(BMI), lipid-profile, fasting plasma glucose(FPG), fasting plasma insulin (FINS), homeostasis model assessment of insulin resistance(HOMA-IR),the serum IL-10, and hs-CRP levels were measured. The concentration of serum IL-10 was measured by enzyme linked immunosorbent assay (ELISA), and serum hs-CRP level by the latex-enhanced immuno- turbidimetric assay.@*RESULTS@#Compared with the control group, the serum IL-10 level in the MS group was significantly lower (P<0.05), and the concentration of hs-CRP was obviously higher (P<0.05). Using Pearson's correlation analysis, the serum IL-10 level was negatively related with HOMA-IR(r=-0.684,P=0.000)and FINS(r=-0.742,P=0.000); hs-CRP was positively related with BMI(r=0.372,P=0.002), HOMA-IR(r=0.276,P=0.021)and FINS(r=0.312,P=0.008)in the MS group. Stepwise regression analysis suggested that FINS might be the influencing factors of IL-10; BMI and FINS might be the influencing factors of hs-CRP in patients with MS.@*CONCLUSION@#In old male patients with MS, the concentration of serum IL-10 decreases, and the serum hs-CRP level increases obviously. This suggests chronic inflammation.
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Anciano , Humanos , Masculino , Persona de Mediana Edad , Proteína C-Reactiva , Metabolismo , Ensayo de Inmunoadsorción Enzimática , Inflamación , Interleucina-10 , Sangre , Síndrome Metabólico , SangreRESUMEN
OBJECTIVE@#To determine the mechanism of Toll-like receptor 4(TLR4) in hypertensive renal injury and the protective effect of fosinopril(Fos) and losartan(Los).@*METHODS@#NRK-52E was incubated into 5 groups: NRK-52E (normal control), NRK-52E+AngII, NRK-52E+AngII+Fos(10(-5) mmol/L),and NRK-52E+AngII+Los(10(-5) mmol/L), NRK-52E +AngII+Fos(10(-5) mmol/L)+Los(10(-5) mmol/L). TLR4-specific RNAi plasmids were stably transfected into NRK-52E. After 24 h, TLR4, IL-6, and TNF-alpha mRNAs were examined by reverse transcription-polymerase chain reaction(RT-PCR). TLR4 proteins were detected by Western blot, NF-kappaB nuclear translocations were tested by immunocytochemistry,and IL-6 and TNF-alpha supernatant levels were tested by enzyme linked immuno-sorbent assay(ELISA).@*RESULTS@#TLR4, NF-kappaB, IL-6,and TNF-alpha were highly expressed in AngII induced NRK-52E(P<0.01). In NRK-52E that was stably transfected TLR4-special RNAi plamids, TLR4 protein and mRNA expression were obviously inhibited(P<0.05). After stimulation by AngII, the TLR4, IL-6, TNF-alpha levels in the stabe transfection group were increased compared with the normal group(P<0.05). Fos or/and Los down-regulated TLR4, IL-6, and TNF-alpha expressions(P<0.05), but no cooperation was observed.@*CONCLUSION@#TLR4 may lead to inflammatory reaction in hypertensive renal injury. Fos or/and Los can decrease the expressions of TLR4 and correlate inflammatory factors, which may be part of the renal protective mechanism.
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Animales , Ratas , Línea Celular , Células Epiteliales , Alergia e Inmunología , Metabolismo , Fosinopril , Farmacología , Hipertensión , Enfermedades Renales , Túbulos Renales , Biología Celular , Metabolismo , Losartán , Farmacología , Interferencia de ARN , ARN Mensajero , Genética , Receptor Toll-Like 4 , GenéticaRESUMEN
The N-terminal segment (FR-1) of the heavy chain (VH) of antibodies may have a great impact on IgG secretion in Escherichia coli and other hosts. Decrease in secretion may be caused by a single amino acid change in the framework region. To investigate the high antibody expression in mammalian cells, we designed the site-directed mutagenesis of the FR-I of the pCMV-RV/VH gene,which expressed the immunoglobulin heavy chain of human anti-Rabies virus antibody. Mutating Glu (H6) to Gln could improve both antibody secretion and affinity. The immunofluorescence assay indicated that both the secretion-deficient antibodies and the secretion- efficient antibodies could be transcribed and translated intracellularly, and led into ER,then transferred to Golgi apparatus,and the difference in secretion may relate to the contribution of the FR-I to the folding and assembly of the antibody. In this study, we have confirmed experimentally that the nature of residues H6 in antibody heavy chains indeed determines the antibody secretion in mammalian cells. These results also provide the basis for antibody production.
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Animales , Humanos , Anticuerpos , Genética , Alergia e Inmunología , Metabolismo , Anticuerpos Antivirales , Genética , Alergia e Inmunología , Metabolismo , Afinidad de Anticuerpos , Transporte Biológico , Células COS , Chlorocebus aethiops , Citomegalovirus , Genética , Retículo Endoplásmico , Metabolismo , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Glicoproteínas , Genética , Alergia e Inmunología , Aparato de Golgi , Metabolismo , Fragmentos de Inmunoglobulinas , Genética , Alergia e Inmunología , Cadenas Pesadas de Inmunoglobulina , Química , Genética , Alergia e Inmunología , Región Variable de Inmunoglobulina , Química , Genética , Alergia e Inmunología , Mutagénesis Sitio-Dirigida , Plásmidos , Genética , Virus de la Rabia , Genética , Alergia e Inmunología , MetabolismoRESUMEN
Two human Fab antibodies against avian influenza A (H5N1) virus were obtained by panning a H5N1 patient-derived antibody phage library using purified virions of the H5N1 patient isolate A/Anhui/1/2005 and HA protein of the H5N1 reference viruse A/Viet Nam/1203/2004. After testing the binding properties and antiviral function to H5N1 virus, the selected Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell system. Both mAbs, AVFluIgG01 and AVFluIgG03, bound to HA in immunofluorescence assay (IFA) without cross-reaction with the other substypes of influenza A viruses (H1N1, H3N2). The cross-reactivity of the two antibodies for different strains of H5N1 was tested in vitro by micro-neutralization assays. In vitro, mAb AVFluIgG01 potently neutralized not only the selected well-characterized Clade 2 H5N1 viruses isolated from mainland of China except A/Guangdong/1/2006, but also the Clade 1 representative isolate A/Viet Nam/1203/2004; and AVFluIgG03 neutralized all the selected Clade 2 H5N1 viruses isolated from mainland of China, but had no neutralizing activity with the Clade 1 H5N1 virus A/Viet Nam/1203/2004. The results bring new prospect for the prophylaxis or treatment of H5N1 virus infection and may provide a clue for novel vaccine development.
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Animales , Humanos , Secuencia de Aminoácidos , Anticuerpos Antivirales , Genética , Alergia e Inmunología , Especificidad de Anticuerpos , Aves , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Subtipo H5N1 del Virus de la Influenza A , Genética , Alergia e Inmunología , Vacunas contra la Influenza , Genética , Alergia e Inmunología , Gripe Aviar , Alergia e Inmunología , Virología , Datos de Secuencia Molecular , Pruebas de Neutralización , Proteínas Recombinantes , Alergia e Inmunología , Homología de Secuencia de AminoácidoRESUMEN
BpHi006A cDNA is 1943 bp in length, and contains one putative open reading frame that is 795 bp long. The expression of BpHi006A was induced by BPH feeding. BpHi006A protein contains a N-terminal domain and a C-terminal domain of glutathione S-transferase, and therefore, it belongs to the superfamily of glutathione S-transferase. BpHi006A protein exhibited 61% amino acid sequence identity to tetrachloro-p-hydroquinone reductive dehalogenase-related protein of Arabidopsis thaliana. Sequence analysis of these two proteins indicates that they belong to a new group of plant GSTs.
Asunto(s)
Animales , Secuencia de Aminoácidos , Clonación Molecular , Genes de Plantas , Glutatión Transferasa , Genética , Hemípteros , Fisiología , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Oryza , Genética , Proteínas de Plantas , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To study the resistance of methicillin-resistant staphylococcus aureus (MRSA), an indicator used in hospitals.</p><p><b>METHODS</b>We used minimal inhibitory concentrations (MIC) of iodoph and chlorhexidine to MRSA, methicillin-sensitive staphylococcus aureus (MSSA) and staphylococcus aureus ATCC6538.</p><p><b>RESULTS</b>Obvious difference between MRSA and MSSA the MIC of Iodophor was noticed. Among MICs, 5.3% MRSA strains were 2-folds and 28.9% MRSA strains were 1.5 fold more than staph. aureus ATCC6538, while the MIC of 11.1% MSSA strains raised 1.5 fold than ATCC6538. The MIC of 83.3% MSSA strains were the same to staph. aureus ATCC6538. The MIC of chlorhexidine to MRSA, MSSA and staphylococcus aureus ATTC6538 were similar to each other.</p><p><b>CONCLUSION</b>Results showed that some MRSA were more resistant to Iodophor than staph. aureus ATCC6538, but remained the same resistance to Chlorhexidine. Thus the concentration of Iodophor should be raised when the resistant strains were isolated.</p>