Clinical diagnostic value of plasma exosomes S100P protein in patients with heatstroke / 解放军医学杂志

Objective: To investigate the plasma exosome S100P levels in patients with heatstroke and evaluate its clinical diagnostic value. Methods: Thirty heatstroke patients were selected from June 2016 to June 2018 in the General Hospital of Southern Theatre Command of PLA, and 15 healthy people in the Physical Examination Center of our hospital were selected as control. The venous blood was collected, followed by plasma exosomes extraction using polyethylene glycol precipitation. The plasma exosome S100P was quantified by ELISA. The correlation analysis between plasma exosomal S100P, clinical indicators, and disease severity was performed. The receiver operating characteristic (ROC) curve was analyzed to evaluate the efficacy of plasma exosome S100P in diagnosing heatstroke. Results: The level of plasma exosomes S100P in patients with heatstroke significantly increased [healthy people vs. heatstroke patients: (195.89 ± 131.97) pg/ml vs. (1163.62 ± 747.15) pg/ml, P<0.001], and the level of plasma exosome S100P was significantly correlated with the severity of heatstroke patients [Mild heatstroke patients vs. severe heatstroke patients (553.63 ± 311.85) pg/ml vs. (1570.28 ± 672.03) pg/ml, P<0.001]. The correlation coefficients r of plasma exosome S100P with APACHE II score, SOFA score and MODS score were 0.888, 0.901, and 0.887, respectively (P<0.001). The area under the ROC curve for the diagnosis of heatstroke by plasma exosomal S100P was 0.931 (95%CI 0.857-1.000, P<0.001). The area under the ROC curve of plasma exosomal S100P for assessing the severity of heatstroke was 0.956 (95%CI 0.890-1.000, P<0.001). Conclusion: The level of plasma exosomal S100P correlates with severity of heatstroke and can be used as a potential marker for diagnosis and prognosis of heatstroke.

Expression and analysis of histo-blood group antigens binding pattern of GⅡ.2 norovirus P protein / 中华实验和临床病毒学杂志

Objective@#During the 2016 winter season, GII.2 norovirus(NoV) suddenly emerged in China. To elucidate its mechanism of epidemic, this study focused on characteristics of binding between the P protein of capsid and histo-blood group antigens (HBGAs).@*Methods@#The research object was GⅡ.2 ZTX strain which had an outbreaks by the end of 2016 in Beijing. Recombinant prokaryotic expression plasmid was constructed, and the expression of virus P protein was determined and purified. The P protein characteristics of binding to HBGAs was studied through saliva and oligosaccharide binding experiments.@*Results@#Soluble P protein was successfully obtained, and combined with type A, B, AB saliva.@*Conclusions@#The result illuminate the combination with new outbreaks of NoV and salivary types, which provided a basis for its pathogenic mechanism and prevention and control measures.

Detection of Human Anti-Trypanosoma cruzi Antibody with Recombinant Fragmented Ribosomal P Protein

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.

Olfactory function and related factors in systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the olfactory function in patients with systemic lupus erythematosus (SLE) and to explore factors that may influence it.Methods The Connecticut Chemosensory Clinical Research Center (CCCRC) test was carried out in SLE patients and healthy controls for olfactory function testing.ELISA method was used to detect anti-ribosomal P protein antibody in the serum.The statistical methods used in this study including t test,ANOVA,LSD-t test,Pearson correlation analysis,multiple linear regression analysis,andx2 test.Results ① The CCCRC scores of the active,inactive group and healthy controls were compared,the difference was statistical significant (F=26.52,P=0.01).CCCRC score in active SLE group (62±16) was lower than that of the inactive group (80±13) the and the normal control group (83±12) (P＜0.01),while there was no statistical significant difference between the inactive group and the normal controls (P=0.226).② CCCRC score was lower in 16 ARPA positive SLE patients (61±17) than 49 negative patients (74±16) (t=2.681,P=0.009).③ In addition,CCCRC score showed a negative correlation with ARPA serum concentration (r=-0.327,P=0.008).There was no significant correlation between CCCRC score and disease course (r=0.141,P＞0.05).④ Multiple linear regression analysis indicated that CCCRC score was associated with age and ARPA.Conclusion SLE patients have olfactory dysfunction and the dys-function is associated with age and ARPA.

Plant defence proteins during aphid infestation.

Parasitism by phloem-feeding insects, such as aphids and whiteflies, are widespread and often serious constraint on plant growth. Aphids successfully exploit a broad range of vascular plants. Despite the ubiquity of phloem feeding insects, in depth knowledge of plant defence and plantmicrobe interactions is still lagging. This review summarizes the current knowledge about the phloem sap proteins that are involved in defence reactions with probable functions in wound and defence reactions.

Structural analysis of human respiratory syncytial virus P protein: identification of intrinsically disordered domains

Human Respiratory Syncytial Virus P protein plus the viral RNA, N and L viral proteins, constitute the viral replication complex. In this report we describe that HRSV P protein has putative intrinsically disordered domains predicted by in silico methods. These two domains, located at the amino and caboxi terminus, were identified by mass spectrometry analysis of peptides obtained from degradation fragments observed in purified P protein expressed in bacteria. The degradation is not occurring at the central oligomerization domain, since we also demonstrate that the purified fragments are able to oligomerize, similarly to the protein expressed in cells infected by HRSV. Disordered domains can play a role in protein interaction, and the present data contribute to the comprehension of HRSV P protein interactions in the viral replication complex.

Prokaryotic expression of P gene from rabies virus and application of the indirect ELISA assay in the detection of its gene products / 中国人兽共患病学报

The complete length of P gene from rabies virus was amplified by RT-PCR using a pair of specific primers designed according to the relevant sequences from GenBank. The PCR product was cloned into cloning expression vestor pGM-T to obtain the cloning expressed plasmid pGM-T-P. After double-digestion by NotI and EcoRI, the product was transferred into prokaryotic expression vetor pET-32a(+)to obtain the prokaryotically expressed plasmid pET-32a-P. The target gene was then expressed in the E.coli BL21(DE3) cell with IPTG induction. The highest expression of target protein was analysed by SDS-PAGE, and the good immunoreactivity to rabies virus antibodies was proved by Western-blot analysis. By using purified protein, the indirect ELISA assay for the detection of rabies virus antibodies in canine serum was applied after management of the optional working condition.

Manifestaciones renales y neuropsiquiátricas en pacientes con lupus eritematoso sistémico y anticuerpos anti-P ribosomal / Renal and neuropsychiatric manifestations in patients with systemic lupus erythematosus and ribosomal P protein autoantibodies

Los auto anticuerpos contra la proteína P ribosomal (anti-P ribosomales) se presentan en aproximadamente el 15% de los pacientes con lupus eritematoso sistémico (LES). Estos anticuerpos fueron inicialmente asociados con psicosis lúpica y enfermedad neuropsiquiátrica. Posteriormente se reconoció su asociación con alto riesgo de compromiso renal y hepático. Objetivos: evaluar la coexistencia serológica y clínica de anticuerpos anti-P ribosomal y anti-DNA en pacientes con LES con compromiso renal y neuropsiquiátrico. Materiales y métodos: casos: 12 pacientes con lupus neuropsiquiátrico (cambios conductuales 6, depresión 4, alucinaciones 3, alteración cognitiva 2, convulsiones 2 y psicosis 2). Controles: 13 pacientes con actividad por LES sin evidencia de manifestaciones neuropsiquiátricas. Todos los pacientes estuvieron activos para el tiempo de la evaluación. Los anticuerpos anti-P ribosomal fueron determinados por ELISA y los anti-dcDNA por el método de Crithidia luciliae. La función renal fue valorada por medición de creatinina y el compromiso renal con uroanálisis. Resultados: la edad media fue de 39 años, 2 hombres / 23 mujeres. Ocho casos (66,6%) y seis (46,1%) controles tenían compromiso renal. El 20% de los pacientes fueron positivos para anti-P ribosomal (5/25 pacientes: 2 controles y 3 casos). El 36% de los pacientes tenían anticuerpos anti-dsDNA (30,7% controles y 41,6% casos). La presencia de anticuerpo P ribosomal estuvo asociada con anti-dcDNA (p = 0,002) y con nefritis lúpica (p = 0,046), pero no hubo asociación entre el anti-P ribosomal y las manifestaciones neuropsiquiátricas (p = 0,645). Conclusiones: la presencia de anticuerpos anti-P ribosomal está asociada con anti-dcDNA y nefritis lúpica. Este estudio no encontró asociación estadísticamente significante entre anti-P ribosomal y manifestaciones neuropsiquiátricas del LES.

Summary The ribosomal P protein autoantibodies are present in approximately 15% of patients with Systemic Lupus Erythematosus (SLE). These antibodies were initially associated with lupus psychosis and neuropsychiatric diseases. Posteriorly, they were associated to a higher risk of renal and liver involvement in patients with SLE. Objective: to evaluate the serologic and clinic coexistence of ribosomal P protein autoantibodies and anti-dsDNA in patients with SLE with renal and neuropsychiatric manifestations. Materials and Methods: cases: 12 patients with neuropsychiatric and renal involvement (behavior changes 6, depression 4, hallucinations 3, cognitive impairment 2, seizures 2 and psychosis 2). Controls: 13 patients with active SLE, without evidence of neuropsychiatric manifestations. SLE was active in all patients during the process of evaluations. The anti-ribosomal P protein antibodies were determined by ELISA and the dsDNA antibodies by Crithidia luciliae´s immunofluorescence test. The renal function was evaluated by serum creatinin and urinalyses. Results: the median age was 39 years, 2 men/23 women. 8 cases (66.6%) and 7 controls (53.8 %) have renal involvement. 20% of patients were positive for anti P-ribosomal antibodies (5/25 patients: 2 controls and 3 cases). 36% of patients have anti dsDNA antibodies (30.7% controls and 41.6% cases). The presence of anti-ribosomal P protein antibodies was associated with anti dsDNA antibodies (p = 0.002) and with lupus nephritis (p = 0.046), but no association between ribosomal P protein autoantibodies and neuropsychiatric manifestations was found (p = 0.645). Conclusions: the P-ribosomal antibodies are associated with anti-dsDNA and lupus nephritis. This study did not show statistically significant associations between ribosomal P protein autoantibodies and neuropsychiatric manifestations in patients with SLE.

Switch of Regulatory Domains of P-protein and T-protein from E. coli / 中国生物化学与分子生物学报

Chorismic acid is a mid-metabolite that plays a central role in the metablism process distributing in the bacterium, epiphyte and plants. It is a common precursor substance of the all aromatic amino acids that can turn into phenylalanine and tyrosine catalyzed by bi-functional enzyme chorismate mutase (CM)-prephenate dehydratase (PDT) and chorismate mutase-prephenate dehydrogenase (PDH) respectively. CMp-PDT with its regulate domain Rp were called P-protein and CMt-PDH with its regulate domain Rt were called T-protein. P-protein and T-protein from E. coli. have a similar structure, both of which contained three domains: CMp, PDT, Rp in P-protein and CMt, PDH, Rt in T-protein. P-protein and T-protein are regulated by their effectors phenylalanine and tyrosine respectively through binding to their Rp and Rt domains. Rp and Rt domains were switched between P-protein and T-protein by cloning of chimeric proteins. The results showed that regulatory effects were switched along the switch of R domains and the switch of the regulatory domains lead to the switch of effectors. It means that the combination of the regulatory domain and the effector is specific and the regulating of the regulatory domain to the enzyme activity is non-specific. This property of R domains may make them possible molecular elements in the study of molecular machines.

Molecular cloning of ribosomal P protein in Toxoplasma gondii and the availability to detect antibody against recombinant protein in toxoplasmosis patients

Among the panel of monoclonal antibodies (mAb) against Toxoplasma gondii, mAb of Tg621 (Tg621) clone blotted 38 kDa protein which localized in the cytoplasm of tachyzoites by immunofluorescence microscopy. The protein was not released into the parasitophorous vacuole during or after invasion. The cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg621. The full length cDNA sequence was completed with 5'-RACE as 1, 592 bp, which contained open reading frame of 942 bp. The deduced amino acid sequence of Tg621 consisted of a polypeptide of 313 amino acids, with significant homology to ribosomal P proteins (RPP) of other organisms especially high to those of apicomplexan species. The expressed and purified TgRPP was assayed in western blot with the sera of toxoplasmosis patients and normal sera, which resulted in the 74.0% of positive reactions in toxoplasmosis patients whereas 8.3% in normal group. Therefore, the antibody formation against TgRPP in toxoplasmosis patients was regarded as specific for T. gondii infection and suggested a potential autoantibody.

Mutagenesis study to ameliorate the bacterial expression of phosphoprotein P of vesicular stomatitis virus.

The phosphoprotein gene of vesicular stomatitis virus, a Rhabdovirus, has been inserted into bacterial expression plasmids containing the Escherichia coli tac promoter and ribosome binding site (RBS). A low level of expression of the protein was detected. Sequence analysis showed the presence of 15 nucleotides in the spacer region i.e., between the Shine- Dalgarno sequence and ATG. Alteration of the distance and the sequence in the spacer region by oligonucleotide-directed mutagenesis revealed a correlation among the expression levels, accessibility of the RBS and requirement for a minimum spacing of at least 7 nucleotides between the Shine-Dalgarno sequence and ATG for optimal gene expression.