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1.
Chinese Journal of Pediatrics ; (12): 154-158, 2023.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-970255

RESUMO

Objective: To investigate the clinical features and gene variation characteristics of children with dynein cytoplasmic 1 heavy chain 1 (DYNC1H1) gene associated spinal muscular atrophy with lower extremity predominant (SMALED) 1. Methods: The clinical data of 4 SMALED1 children admitted to Peking University First Hospital from December 2018 to May 2021, who were found to have pathogenic variation of DYNC1H1 gene through genetic testing, except for other genes known to be related to motor retardation, were retrospectively summarized to analyze the phenotype and genotype characteristics. Results: There were 3 males and 1 female. The age of onset was 1 year, 1 day, 1 day and 4 months, respectively. The age of diagnosis was 4 years and 10 months, 9 months, 5 years and 9 months, and 3 years and 1 month, respectively. The clinical manifestations were muscle weakness and muscular atrophy of lower limbs, 2 cases with foot deformity, 1 case with early non progressive joint contracture, 1 case with hip dislocation and 1 case with mental retardation. De novo heterozygous missense variations in DYNC1H1 gene were found in all 4 children. According to the rating of American College of medical genetics and genomics, they were all possible pathogenic and pathogenic variations, with p.R598C, p.P776L, p.Y1109D variations had been reported, and p.I1086R variation had not been reported. Conclusions: For those with unexplained lower limb muscle weakness, muscle atrophy, joint contracture and foot deformity, upper limb motor ability related retention, with or without mental retardation, as well as the motor ability progresses slowly, it is necessary to consider the possibility of SMALED1 and the detection of DYNC1H1 gene when necessary.


Assuntos
Feminino , Masculino , Humanos , Deficiência Intelectual , Estudos Retrospectivos , Atrofia Muscular Espinal/genética , Extremidade Inferior , Debilidade Muscular , Atrofia Muscular , Contratura , Dineínas do Citoplasma/genética
2.
Chinese Journal of Pediatrics ; (12): 345-350, 2023.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-985874

RESUMO

Objective: To analyze the clinical and genetic characteristics of pediatric patients with dual genetic diagnoses (DGD). Methods: Clinical and genetic data of pediatric patients with DGD from January 2021 to February 2022 in Peking University First Hospital were collected and analyzed retrospectively. Results: Among the 9 children, 6 were boys and 3 were girls. The age of last visit or follow-up was 5.0 (2.7,6.8) years. The main clinical manifestations included motor retardation, mental retardation, multiple malformations, and skeletal deformity. Cases 1-4 were all all boys, showed myopathic gait, poor running and jumping, and significantly increased level of serum creatine kinase. Disease-causing variations in Duchenne muscular dystrophy (DMD) gene were confirmed by genetic testing. The 4 children were diagnosed with DMD or Becker muscular dystrophy combined with a second genetic disease, including hypertrophic osteoarthropathy, spinal muscular atrophy, fragile X syndrome, and cerebral cavernous malformations type 3, respectively. Cases 5-9 were clinically and genetically diagnosed as COL9A1 gene-related multiple epiphyseal dysplasia type 6 combined with NF1 gene-related neurofibromatosis type 1, COL6A3 gene-related Bethlem myopathy with WNT1 gene-related osteogenesis imperfecta type XV, Turner syndrome (45, X0/46, XX chimera) with TH gene-related Segawa syndrome, Chromosome 22q11.2 microduplication syndrome with DYNC1H1 gene-related autosomal dominant lower extremity-predominant spinal muscular atrophy-1, and ANKRD11 gene-related KBG syndrome combined with IRF2BPL gene-related neurodevelopmental disorder with regression, abnormal movement, language loss and epilepsy. DMD was the most common, and there were 6 autosomal dominant diseases caused by de novo heterozygous pathogenic variations. Conclusions: Pediatric patients with coexistence of double genetic diagnoses show complex phenotypes. When the clinical manifestations and progression are not fully consistent with the diagnosed rare genetic disease, a second rare genetic disease should be considered, and autosomal dominant diseases caused by de novo heterozygous pathogenic variation should be paid attention to. Trio-based whole-exome sequencing combining a variety of molecular genetic tests would be helpful for precise diagnosis.


Assuntos
Humanos , Anormalidades Múltiplas , Estudos Retrospectivos , Deficiência Intelectual/genética , Doenças do Desenvolvimento Ósseo/complicações , Anormalidades Dentárias/complicações , Fácies , Distrofia Muscular de Duchenne/complicações , Atrofia Muscular Espinal/complicações , Proteínas de Transporte , Proteínas Nucleares
3.
Arch. esp. urol. (Ed. impr.) ; 75(7): 618-623, 28 sept. 2022. tab, graf
Artigo em Inglês | IBECS | ID: ibc-212084

RESUMO

Objectives: Flexible ureteroscopic lithotripsy (FURL), as a common method for treating upper urinary tract calculi, has the risks of complications such as infection and bleeding. Especially, systemic inflammatory response syndrome (SIRS) after FURL may induce multiple organ dysfunction threatening the lives of patients. We aimed to investigate the clinical characteristics and risk factors of SIRS after FURL. Methods: A total of 157 upper urinary tract calculus patients treated with FURL from January 2018 to December 2019 were enrolled, and clinical outcomes and complications were analyzed. Patients were divided into SIRS group (n = 31) and non-SIRS group (n = 126) according to the presence or absence of SIRS after FURL. Their clinical data were compared by univariate analysis, and the factors with statistically significant difference were incorporated into LASSO logistic regression analysis. The model was visualized using a nomogram, and model discrimination and accuracy were verified. Results: The results of univariate analysis indicated that there were significant differences in gender, average stone size, preoperative urinary white blood cell count, surgery time and postoperative stone bacterial culture between the two groups. The results of LASSO logistic regression analysis showed that the above factors were independent risk factors for patients with SIRS. The C-index of the SIRS risk prediction model was 0.992. The area under the ROC curve of this model was 0.944 (95% CI: 0.913-0.997), the sensitivity was 97.9%, and the specificity was 95.8%. The average absolute error between actual and predicted risk probabilities was 0.028. The model for predicting the risk of SIRS had good discrimination and high consistency with the actual observed value (AU)


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Síndrome de Resposta Inflamatória Sistêmica/etiologia , Cálculos Renais/cirurgia , Litotripsia/efeitos adversos , Ureteroscopia/efeitos adversos , Estudos Retrospectivos , Fatores de Risco
4.
Artigo em Inglês | MEDLINE | ID: mdl-36688184

RESUMO

The Omicron variant caused a surge of infections with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Viet Nam in early 2022, signalling community transmission. We report on active whole-genome sequencing surveillance of positive SARS-CoV-2 samples collected at that time in northern Viet Nam from international arrivals and community clusters. We used an amplicon protocol developed with 14 polymerase chain reaction products and the Illumina iSeq 100 platform. Overall, 213 nasopharyngeal or throat swabs were analysed, of which 172 samples were identified with the Omicron variant. Of these, 80 samples were collected from community cases in February 2022, among which 59 samples were sublineage BA.2 and one sample was the recombinant XE variant. Our results indicated that Omicron had replaced Delta as the dominant variant in a very short period of time and that continuously conducting active whole-genome sequencing surveillance is necessary in monitoring the evolution and genomic diversity of SARS-CoV-2 in Viet Nam.


Assuntos
COVID-19 , Humanos , COVID-19/epidemiologia , SARS-CoV-2/genética , Vietnã/epidemiologia , Genômica
5.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-922356

RESUMO

PURPOSE@#To investigate the clinical value of urine interleukin-18 (IL-8), neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) for the early diagnosis of acute kidney injury (AKI) in patients with ureteroscopic lithotripsy (URL) related urosepsis.@*METHODS@#A retrospective study was carried out in 157 patients with urosepsis after URL. The patients were divided into AKI group and non-AKI group according to the Kidigo guideline and urine IL-8, NGAL and KIM-1 levels were detected by enzyme-linked immunosorbent assay at 0, 4, 12, 24 and 48 h after the surgery. Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic value of these three biomarkers for postoperative AKI.@*RESULTS@#The level of urine IL-8, NGAL and KIM-1 in AKI group was significantly higher than that in non-AKI group at 4, 12, 24 and 48 h (p < 0.01). The ROC analysis showed the combined detection of urine IL-8, NGAL and KIM-1 at 12 h had a larger area under curve (AUC) than a single marker (0.997, 95% CI: 0.991-0.998), and the sensitivity and specificity were 98.2% and 96.7%, respectively. Pearson correlation analysis showed that the levels of urine NGAL at 4, 12, 24 and 48 h in AKI patients were positively correlated with the levels of urine KIM-1 and IL-18 (p < 0.01).@*CONCLUSION@#AKI could be quickly recognized by the elevated level of urine IL-8, NGAL and KIM-1 in patients with URL-related urosepsis. Combined detection of the three urine biomarkers at 12 h after surgery had a better diagnostic performance, which may be an important reference for the early diagnosis of AKI.


Assuntos
Humanos , Injúria Renal Aguda/etiologia , Biomarcadores , Diagnóstico Precoce , Receptor Celular 1 do Vírus da Hepatite A , Interleucina-18 , Interleucina-8 , Lipocalina-2 , Litotripsia , Estudos Retrospectivos , Ureteroscopia
6.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-1031798

RESUMO

@#The Omicron variant caused a surge of infections with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Viet Nam in early 2022, signalling community transmission. We report on active whole-genome sequencing surveillance of positive SARS-CoV-2 samples collected at that time in northern Viet Nam from international arrivals and community clusters. We used an amplicon protocol developed with 14 polymerase chain reaction products and the Illumina iSeq 100 platform. Overall, 213 nasopharyngeal or throat swabs were analysed, of which 172 samples were identified with the Omicron variant. Of these, 80 samples were collected from community cases in February 2022, among which 59 samples were sublineage BA.2 and one sample was the recombinant XE variant. Our results indicated that Omicron had replaced Delta as the dominant variant in a very short period of time and that continuously conducting active whole-genome sequencing surveillance is necessary in monitoring the evolution and genomic diversity of SARS-CoV-2 in Viet Nam.

7.
Chinese Pharmacological Bulletin ; (12): 1132-1136, 2022.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1014024

RESUMO

Connexin (Cx), a multigene-encoded transmembrane protein family, forms either gap junctions ( GJ) or hemichannels (HC) to mediate intercellular communication in plasma mem¬brane between adjacent cells or interacts with proteins by its car- boxyl terminal in the cytoplasm to participate in the process of tumor cell proliferation, apoptosis, necrosis, invasion, metasta¬sis, drug resistance and stem cell characteristics.However, mi- slocalization of Cx in cytoplasm or nucleus often occurs in many tumors, and involved in the occurrence and development of tumors.Subcellular localization of Cx is affected by post-transla- tional modifications, including phosphorylation, ubiquitination, and acetylation.In this paper the classification and function of Cx, the relationship between subcellular localization of Cx and tumorigenesis and the regulation of post-translational modifica¬tion on Cx are reviewed in order to provide new ideas for the study of Cx as a potential target for cancer therapy.

8.
Chinese Pharmacological Bulletin ; (12): 1023-1031, 2022.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1014058

RESUMO

Aim To explore the apoptosis of small eell lung eancer ( SCLC ) eells HI688 and H446 induced by nitidine chloride and its possible mechanism.Methods The effect of nitidine chloride or cisplatin ( DDP ) on the activity of SCLC cells was detected by j J MTT method; the morphological changes of cells trea¬ted with nitidine chloride or DDP were observed by in- verted fluorescence microscope and HE staining; the effect of nitidine chloride or DDP on apoptosis was de¬tected by flow cytometry; the effect of apoptosis inhibi¬tor Z-VAD-FMK on apoptosis induced by nitidine chlo¬ride or DDP was detected by MTT method.The expres¬sions of Bax , Bcl-2, caspase-3 , PARP, p-PI3K and p- Akt in the cells treated with nitidine chloride or DDP were detected by Western blot.Results MTT results showed that the viability of SCLC cells was significantly reduced after 48 hours of treatment with nitidine chlo¬ ride; compared with DDP, nitidine chloride could in¬hibit SCLC cells with less IC50; inverted fluorescence microscope and HE staining showed that nitidine chlo¬ride could induce apoptosis in SCLC cells, similar to DDP; flow cytometry showed that nitidine chloride J J could induce apoptosis in SCLC cells.The results of MTT assay showed that the inhibitory effect of nitidine chloride on apoptosis of SCLC cells could be partially antagonized by apoptosis inhibitor Z-VAD-FMK.West¬ern blot results showed that, similar to DDP, nitidine chloride could inhibit the expression of PI3K and Akt, increase Bax, inhibit Be 1-2, and promote the cleavage of caspase-3 and PAH P.Conclusion Nitidine chlo¬ride can induce apoptosis of SCLC cells by inhibiting the activation of P13K and Akt.

9.
Chinese Pharmacological Bulletin ; (12): 768-774, 2021.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1014434

RESUMO

Aim To study the protective effect of ZN-RF2 on OGD/R-induced injury and the autophagy-related mechanism in PC12 cells. Methods PC12 cells were cultured in vitro and divided into normal group and OGD/R group. qRT-PCR and Western blot were used to measure the mRNA and protein expressions of ZNRF2. To explore the effect of ZNRF2 on OGD/R-induced injury in PC12 cells, cells were grouped into normal group, OGD/R group, LV-ZNRF2 group, LV-NC group, siR-ZNRF2 group and siNC group. Cell viability was detected by MTT assay, cell apoptosis was measured by flow cytometry and the expressions of autophagy-related proteins LC3II, p62, Beclin-l were accessed by Western blot. Results Compared with normal group, the cell viability decreased in OGD/R group, the cell apoptosis increased markedly, and the expressions of ZNRF2 mRNA and protein were downregulated significantly. Simultaneously, the proteins expressions of LC3II and Beclin-1 increased, and the expression of p62 protein decreased in OGD/R group. Compared with OGD/R group, the cell viability was enhanced, the cell apoptosis and autophagy were decreased in LV-ZNRF2 group. In contrast, the cell viability decreased and the cell apoptosis and autophagy were aggravated after transfecting siR-ZNRF2. Conclusions ZNRF2 protects PCI2 cells from the injury caused by OGD/R and its mechanism may be related to the inhibition of autophagy.

10.
China Pharmacy ; (12): 1077-1082, 2021.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-876580

RESUMO

OBJECTIVE:To study the anti- inflammatory effect and mechanism of Jingulian capsule on inflammatory model rats. METHODS :Totally 48 rats were randomly divided into blank control group ,model group ,Jingulian capsule low-dose , medium-dose and high-dose groups (0.66,1.32,2.64 g/kg),dexamethasone group (positive control ,0.945 mg/kg),with 8 rats in each group. Blank control group and model group were given constant volume of water intragastrically ,and other groups were given relevant medicine intragastrocally ,twice a day ,for consecutive 3 days. Thirty minutes after last administration ,model group and administration groups were given lipopolysaccharide (10 mg/kg)intraperitoneally to induce inflammatory model. Six hours after intraperitoneal injection ,the contents of TNF-α,IL-1β,IL-6,PGE2 in serum were detected by ELISA. The wet to dry weight (W/D)ratio of lung tissue were determined. HE staining was used to observe the pathological changes of lung tissue . RT-PCR was used to detect the mRNA expression of TNF-α,IL-6,PGE2 and IL- 1β in lung tissue. Western blot assay was used to detect the phosphorylation of NF-κB p65 protein and the expression of IκBα protein in lung tissue. RESULTS:Compared with blank ; control group ,the contents of TNF-α,IL-1β,IL-6 and PGE 2 in serum ,the W/D ratio of lung tissue ,the expression of TNF-α,IL-1β,IL-6 and PGE 2 mRNA and the phosphorylation level of NF-κB p65 protein in lung tissue of model group weresignificantly increased ,and the expression of IκBα proteinwas significantly decreased (P<0.05 or P<0.01);a large number of alveolar atrophy and collapse ,alveolar wall thickening ,lung consolidation ,and a large number of inflammatory cell infiltration could be seen in lung tissue. Compared with model group ,the contents of TNF-α,IL-1β(except for low-dose group ), IL-6 and PGE 2 in serum ,as well as the expression of TNF-α(except for high-dose group ),IL-1β,IL-6(except for low-dose , high-dose groups )and PGE 2 mRNA in lung tissue were decreased significantly in Jingulian capsule groups (P<0.05 or P<0.01); the W/D ratio of lung tissue was decreased significantly in Jingulian high-dose group (P<0.05 or P<0.01);the expression of phosphorylation level of NF-κB p65 protein in lung tissue of Jingulian medium-dose group were decreased significantly (P<0.05 or P<0.01),while the expression of IκBα protein was increased significantly(P<0.05);the alveolar structure was clear ,the alveolar wall was slightly thickened , and a small amount of inflammatory cell infiltration was seen in lung tissue. CONCLUSIONS:Jingulian capsule has good anti-inflammatory effect on inflammatory model rats ,the mechanism of which may be related to the inhibition of NF-κB signal pathway.

11.
Protein & Cell ; (12): 120-130, 2019.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-757977

RESUMO

Assembly of eukaryotic ribosome is a complicated and dynamic process that involves a series of intermediates. It is unknown how the highly intertwined structure of 60S large ribosomal subunits is established. Here, we report the structure of an early nucleolar pre-60S ribosome determined by cryo-electron microscopy at 3.7 Å resolution, revealing a half-assembled subunit. Domains I, II and VI of 25S/5.8S rRNA pack tightly into a native-like substructure, but domains III, IV and V are not assembled. The structure contains 12 assembly factors and 19 ribosomal proteins, many of which are required for early processing of large subunit rRNA. The Brx1-Ebp2 complex would interfere with the assembly of domains IV and V. Rpf1, Mak16, Nsa1 and Rrp1 form a cluster that consolidates the joining of domains I and II. Our structure reveals a key intermediate on the path to establishing the global architecture of 60S subunits.

12.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-704239

RESUMO

There were 5 elderly patients with Griesinger syndrome.There was no specificity in the clinical manifestations of Griesinger syndrome in the elderly.The positive rate of stool hookworm eggs was low,but the gastroscopy could help the diagno-sis.

13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-702224

RESUMO

Objective To explore the relationship of impaired brain function after emergency inhalation anesthesia of sevoflurane com -bined with nitrous oxide and cyclic AMP response element binding protein(CREB)signal pathway.Methods A total of 442 patients who were admitted into our hospital from January 2014 to April 2017 were selected as the object of this study.And these patients were divided into 3 groups according to different anesthesia ways,namely the inhalation anesthesia group(118 cases),the local anesthesia group(206 cases), and the control group(118 cases).The blood cyclic adenosine monophosphate(cAMP)concentration were examined by the mini mental state examination(MMSE)score scale.At the same time,the 60 rats were randomly divided into the anesthesia group(n=30)and the con-trol group(n=30).The learning and memory function of the two groups were evaluated by Morris water maze test,and the expressions of cAMP and CREB were measured.Results The blood cAMP concentration and MMSE score in the inhalation anesthesia group were signifi -cantly decreased after inhalation anesthesia(P<0.05).In the place navigation test,rats of the anesthesia group cost much more time to find the platform compared with rats in the control group, and rats of the anesthesia group encountered less times of crossing the platform com-pared with rats in the control group,and the difference was significant(P<0.05).Western blot showed that patients of the anesthesia group had lower cAMP and expression of p-CREB protein,with significant difference(P<0.05).Conclusion Brain function decline after sevoflu-rane inhalation anesthesia combined with nitrous oxide may induced by increasing the nucleus of the second messenger CAMP /Ca2+pathway and decreasing the expression of CREB.

14.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 35(3): 264-268, 2017 Jun 01.
Artigo em Chinês | MEDLINE | ID: mdl-28675010

RESUMO

OBJECTIVE: To explore the therapeutic effect of enhancer of Zeste homolog 2 (EZH2) inhibitor GSK343 on periodontitis by regulating microphage differentiation. METHODS: Macrophage RAW264.7 cells were divided into the blank (A group), control (B group), lipopolysaccharide (LPS) stimulation (C group), and LPS+GSK343 (D group) groups. Phenotype transformations was determined through Western blot analysis and enzyme-linked immunosorbent assay by detecting the differentiation of phenotypic biological markers, including tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), interleukin-10 (IL-10), and Arginase-1 (Arg-1). Metergasis was identified by performing a phagocytosis test on Escherichia coli (E. coli). RESULTS: Macrophage RAW264.7 cells produced classical phenotypic biomarkers (M1) TNF-α and iNOS under LPS stimulation. The expression levels of IL-10 and Arg-1 increased after adding GSK343 into the culture medium. GSK343 also induced the conversion of M1 macrophages into M2 macrophages. Macrophage RAW264.7 cells exerted a phagocytic effect on E. coli, and this effect was enhanced after adding LPS into the culture medium. GSK343 regulated the macrophage RAW264.7 phagocytosis of E. coli. CONCLUSIONS: GSK343 possibly participates in the regulation of macrophage differentiation and, consequently, in the latent treatment of periodontitis.


Assuntos
Proteína Potenciadora do Homólogo 2 de Zeste , Inibidores Enzimáticos/farmacologia , Indazóis/farmacologia , Macrófagos , Periodontite , Piridonas/farmacologia , Arginase , Diferenciação Celular , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Interleucina-10 , Lipopolissacarídeos , Óxido Nítrico Sintase Tipo II , Fagocitose , Fator de Necrose Tumoral alfa
15.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 30(1): 108-109, 2017 Nov 22.
Artigo em Chinês | MEDLINE | ID: mdl-29536723

RESUMO

There were 5 elderly patients with Griesinger syndrome. There was no specificity in the clinical manifestations of Griesinger syndrome in the elderly. The positive rate of stool hookworm eggs was low, but the gastroscopy could help the diagnosis.


Assuntos
Ancilostomíase/diagnóstico , Idoso , Ancylostomatoidea , Animais , Fezes , Humanos , Contagem de Ovos de Parasitas , Sensibilidade e Especificidade
16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-664150

RESUMO

Objective To establish a method for specific gene editing of the second exon of mouse MAD2L1 gene by CRISPR/Cas9, and analyze its off-target effect. Methods The gene editing site for MAD2L1 gene was designed by CHOP?CHOP, and the Cas9?MAD2L1 vector was constructed based on the designed editing site. Cas9?MAD2L1 was then transfected into NIH/3T3 cells and screened with puromycin, followed by observing GFP expression using fluorescence microscopy. The genomic DNA from transfected cells was extracted and a partial fragment of MAD2L1 gene was amplified by PCR. T7E1 analy?sis and Sangger sequencing were used for gene editing and off?target analysis. Results After Cas9?MAD2L1 transfection and puromycin screening, a large number of GFP?expressing cells were observed under the fluorescence microscope. Combined the PCR result with TE71 analysis, the amplified 228 bp PCR products can be digested into 166 bp and 62 bp fragments. The se?quencing result showed that the second exon of MAD2L1 gene was successfully edited, and the off?target effect was undetected in our system. Conclusions The method for specific gene editing of the second exon of mouse MAD2L1 gene by CRISPR/Cas9 is successfully established, and off?target effect of MAD2L1 gene is not detected.

17.
Protein & Cell ; (12): 187-200, 2016.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-757153

RESUMO

The human Shwachman-Diamond syndrome (SDS) is an autosomal recessive disease caused by mutations in a highly conserved ribosome assembly factor SBDS. The functional role of SBDS is to cooperate with another assembly factor, elongation factor 1-like (Efl1), to promote the release of eukaryotic initiation factor 6 (eIF6) from the late-stage cytoplasmic 60S precursors. In the present work, we characterized, both biochemically and structurally, the interaction between the 60S subunit and SBDS protein (Sdo1p) from yeast. Our data show that Sdo1p interacts tightly with the mature 60S subunit in vitro through its domain I and II, and is capable of bridging two 60S subunits to form a stable 2:2 dimer. Structural analysis indicates that Sdo1p bind to the ribosomal P-site, in the proximity of uL16 and uL5, and with direct contact to H69 and H38. The dynamic nature of Sdo1p on the 60S subunit, together with its strategic binding position, suggests a surveillance role of Sdo1p in monitoring the conformational maturation of the ribosomal P-site. Altogether, our data support a conformational signal-relay cascade during late-stage 60S maturation, involving uL16, Sdo1p, and Efl1p, which interrogates the functional P-site to control the departure of the anti-association factor eIF6.


Assuntos
Humanos , Cristalografia por Raios X , GTP Fosfo-Hidrolases , Química , Metabolismo , Domínios Proteicos , Subunidades Ribossômicas Maiores de Eucariotos , Química , Metabolismo , Saccharomyces cerevisiae , Química , Metabolismo , Proteínas de Saccharomyces cerevisiae , Química , Metabolismo
18.
Protein & Cell ; (12): 878-887, 2016.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-757348

RESUMO

Mechanistic target of rapamycin (mTOR) complex 1 (mTORC1) integrates signals from growth factors, cellular energy levels, stress and amino acids to control cell growth and proliferation through regulating translation, autophagy and metabolism. Here we determined the cryo-electron microscopy structure of human mTORC1 at 4.4 Å resolution. The mTORC1 comprises a dimer of heterotrimer (mTOR-Raptor-mLST8) mediated by the mTOR protein. The complex adopts a hollow rhomboid shape with 2-fold symmetry. Notably, mTORC1 shows intrinsic conformational dynamics. Within the complex, the conserved N-terminal caspase-like domain of Raptor faces toward the catalytic cavity of the kinase domain of mTOR. Raptor shows no caspase activity and therefore may bind to TOS motif for substrate recognition. Structural analysis indicates that FKBP12-Rapamycin may generate steric hindrance for substrate entry to the catalytic cavity of mTORC1. The structure provides a basis to understand the assembly of mTORC1 and a framework to characterize the regulatory mechanism of mTORC1 pathway.


Assuntos
Humanos , Linhagem Celular , Microscopia Crioeletrônica , Métodos , Alvo Mecanístico do Complexo 1 de Rapamicina , Complexos Multiproteicos , Química , Estrutura Quaternária de Proteína , Serina-Treonina Quinases TOR , Química
19.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-499877

RESUMO

Objective To evaluate the correlation between the expression of C5b-9 and the gastric carcinoma.Methods In this study, the human gastric adenocarcinoma (HGAC)tissues (n =32 cases)from 32 patients were evaluated by immunohistochemistry and tissue chip.The result was analyzed and evaluated by Multinomial logistic regression and likelihood ratio.Results Multinomial logistic regression and likelihood ratio testing showed that over-expression of C5b-9 in HGAC tissue was significantly correlated with clinical stage (P =0.007) and tumor stage (P =0.005),but not with tumor metastasis,lymphoid nodal status,sex or age.Conclusion C5b-9 can be the criteria of di-agnosis and clinical staging for gastric adenocarcinoma,which may help in diagnosis and assessment disease severity of human gastric carcinoma.

20.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-330251

RESUMO

<p><b>OBJECTIVE</b>This study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.</p><p><b>METHOD</b>The HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.</p><p><b>RESULT</b>The results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.</p><p><b>CONCLUSION</b>This method is able to be a scientific basis of quality assessment according to its convenient and reliable.</p>


Assuntos
Cromatografia Líquida de Alta Pressão , Métodos , Medicamentos de Ervas Chinesas , Inula , Química
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