Construction of recombinant adenovirus expression vector of avian reovirus σC protein and its effect on proliferation of hepatocellular carcinoma cells / 中国生物制品学杂志

@#Objective To construct the recombinant adenovirus vector pAd-σC for the expression of avian reovirus(ARV)aC protein and to detect its effects on the proliferation of hepatocellular carcinoma cells,in order to build up a basis for the development of novel anti-tumor vaccines.Methods The recombinant shuttle vector pShuttle-σC was constructed by PCR amplification of ARV σC gene,and then transformed into competent BJ5183 cells containing the adenovirus vector pAdessy-1.The recombi-nant adenovirus vector pAd-σC was obtained by homologous recombination,and the virus was packaged in HEK293 cells.The virus titer was measured by TCID_(50),the expression of σC protein was determined by Western blot and ELISA,and the effect of virus on the proliferation of human hepatocellular carcinoma cells SMMC7721 was detected by CCK-8 assay.Results The recombinant shuttle vector pShuttle-σC was confirmed to be constructed correctly by double enzyme digestion and sequen-cing,and the recombinant adenovirus vector pAd-σC was constructed correctly as identified by colony PCR.σC protein was successfully expressed in hepatocellular carcinoma cells SMMC7721.The recombinant adenovirus Ad-σC had a titer of 10~(7.5)/0.1 mL,which inhibited the proliferation of hepatocellular carcinoma cells SMMC7721.Conclusion The recombinant adenovirus vector pAd-trC containing ARV σC gene was successfully constructed,and its inhibitory effect on tumor cell proliferation was preliminarily analyzed,which lays a foundation for revealing the molecular mechanism of ARV oncolytic effect and further developing novel anti-tumor biological preparation.

Predictive value of sCTLA-4 and RAD51C proteins for recurrence after interventional therapy for cervical cancer patients / 中华内分泌外科杂志

Objective:To analyze the predictive value of soluble cytotoxic T lymphocyte-associated antigen 4 (sCTLA-4) and RAD51 paralogous gene C (RAD51C) protein in the recurrence of cervical cancer patients after interventional therapy.Methods:A total of 107 patients with cervical cancer who underwent interventional surgery in our hospital from May. 2015 to Apr. 2019 were selected as the research group. postoperative recurrence were recorded. Another 107 patients with benign cervical disease during the same period were selected as the control group. The protein expressions of sCTLA-4 and RAD51C were compared between the two groups and patients with or without recurrence. Logistic regression was used to analyze the influencing factors of postoperative recurrence of cervical cancer patients, and a nomogram model of postoperative recurrence of cervical cancer patients was constructed and verified by calibration curve. The postoperative recurrence rate of cervical cancer patients with different sCTLA-4 and RAD51C protein expressions was compared.Results:The level of sCTLA-4 and the high expression rate of RAD51C protein in the study group were higher than those in the control group ( P<0.05). High-risk human papillomavirus positive, vascular infiltration, interstitial infiltration ≥1/2, paracterine infiltration, high expression of RAD51C protein and high SCTLA-4 level were independent risk factors for postoperative recurrence of cervical cancer ( P<0.05). High-risk human papillomavirus, vascular invasion, interstitial invasion, parametrial invasion, RAD51C protein and sCTLA-4 levels were used to construct a nomogram prediction model for postoperative recurrence of cervical cancer patients. The consistency indices were 0.610 (95% CI: 0.511-0.702), 0.616 (95% CI: 0.517-0.708), 0.640 (95% CI: 0.541-0.730), 0.609 (95% CI: 0.510-0.702), 0.728 (95% CI ranged from 0.633 to 0.809), 0.817 (95% CI ranged from 0.731 to 0.885), and the calibration curve validation showed high consistency. The net benefit rate of combined detection of sCTLA-4 and RAD51C proteins was greater than that of single detection. Conclusions:sCTLA-4 and RAD51C proteins are highly expressed in cervical cancer patients, and the high expression of both indicates that cervical cancer patients have a higher risk of recurrence after surgery. Clinically, the detection of sCTLA-4 and RAD51C protein expression can be used to screen patients with high recurrence risk.

Neutrophil to lymphocyte ratio and C- protein level as markers to predict deterioration in stable COVID-19 patients

The new Corona virus disease (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). It is a highly communicable infectious disease [1]. Clinical manifestations of COVID-19 vary from asymptomatic to ARDS leading to death, it is important to differentiate between severe and non-severe cases. Early warning signs can be identified by several laboratory tests which are often expensive, time taking. Neutrophil-lymphocyte ratio (NLR), C- reactive protein (CRP) are two simple tests that can be used as markers for clinical outcome in COVID-19 patients. The aim and objectives of the study were to correlate patients clinical severity and CRP, NLR levels and to assess direct correlation between CRP levels and NLR. 101 patients who fulfilled the inclusion criteria were taken into the study. Patients are divided into mild, moderate severity based on WHO criteria and CBP, CRP were sent on the 6th/ 7th day of illness. Among the 101 patients, 93 were in mild group, 8 were in moderate group. The study group included subjects who aged between 18 years to 80 years of age. Plasma CRP levels were higher in moderate cases than in mild cases, and this difference was significant (p < 0.001). The mean NLR of moderate severity were significantly higher than those of mild cases (p-0.00003). Results showed that NLR was positively correlated with CRP levels.NLR and CRP are potential, reliable and easy-to-use predictors for deteriorating covid-19 infection. The integration of NLR and CRP may lead to improved predictions and help to triage patients at the time of hospital admission.

Identification and Expression Analysis of PP2C Gene Family Members in Cannabis sativa / 中国实验方剂学杂志

ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp. MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively. ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution. ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.

Identification and Expression Analysis of PP2C Gene Family Members in Cannabis sativa / 中国实验方剂学杂志

ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp. MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively. ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution. ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.

Sex Difference in the Effect of Body Mass Index and Stress on High-Risk Diabetes Mellitus in Korean Adults

PURPOSE: This study aimed to investigate sex differences in the effect of body mass index (BMI) and stress on high-risk diabetes mellitus (DM). METHODS: Secondary analysis of data from 4,271 male and female adults participating in the Sixth Korea National Health and Nutrition Examination Survey 2015 was performed. The participants were evaluated using questionnaires and blood tests. Data were analyzed using descriptive statistics, t-test, χ2-test, and multiple logistic regression analysis (SPSS 24.0). RESULTS: To identify sexspecific effects, interaction variables were included. Hemoglobin A1c (HbA1c) level was higher in men than in women, and the risk of DM decreased 0.31 times in women compared to that in men. As age increased, the odds of risk DM increased 1.03. The risk of DM increased 1.99 times in overweight individuals and 2.79 times for obese individuals compared to that in individuals with normal weight. Stress levels were higher in women than in men, but stress is not an influential factor in high-risk DM. In age-sex interaction, the odds of risk DM increased 1.02 in women compared to that in men as age increased. CONCLUSION: HbA1c level was affected by age-sex interaction, and age and sex should be considered in the application of HbA1c in the diagnosis of DM.

Deficiencia congénita de proteína C en un recién nacido con trombosis y necrosis de tejidos extensa / Congenital C protein deficiency in a newborn with extensive thrombosis and necrosis of tissues

Uno de los trastornos hematológicos más graves del período neonatal es la deficiencia congénita de proteína C, de presentación muy rara, y causa de enfermedad tromboembólica severa y púrpura fulminante en recién nacidos. Se puede sintetizar como una entidad clínico-patológica, de aparición aguda, con trombosis de la vasculatura de la dermis, lo cual conduce a necrosis hemorrágica y progresiva de la piel, asociada a coagulación intravascular diseminada y hemorragia perivascular, que ocurre en el período neonatal. El paciente presentado exhibe los elementos clínico-patológicos que caracterizan la púrpura fulminante, cuyo origen se debe a una deficiencia hereditaria de proteína C, lo cual condujo a la aparición de complicaciones trombóticas severas(AU)

One of the most serious hematological disorders of the neonatal period is congenital C protein deficiency of very rare occurrence and the main cause of severe thromboembolic disease and purpura fulminans in newborns. It may be summarized as a clinical and pathological entity of acute occurrence, with dermis vasculature thrombosis that leads to progressive hemorrhagic necrosis of the skin, associated to disseminate intravascular coagulation and perivascular hemorrhage in the neonatal period. The patient of this report showed the clinical and pathological elements characterizing purpura fulminans the origin of which is due to hereditary C protein deficiency that led to onset of severe thrombotic complications in this patient(AU)

Risk Factors for the Development and Progression of Diabetic Kidney Disease in Patients with Type 2 Diabetes Mellitus and Advanced Diabetic Retinopathy

BACKGROUND: Some patients with type 2 diabetes mellitus (T2DM) do not develop diabetic kidney disease (DKD) despite the presence of advanced diabetic retinopathy (DR). We aimed to investigate the presence of DKD and its risk factors in patients with T2DM and advanced DR. METHODS: We conducted a cross-sectional study in 317 patients with T2DM and advanced DR. The phenotypes of DKD were divided into three groups according to the urine albumin/creatinine ratio (uACR, mg/g) and estimated glomerular filtration rate (eGFR, mL/min/1.73 m²): no DKD (uACR <30 and eGFR ≥60), non-severe DKD (uACR ≥30 or eGFR <60), and severe DKD (uACR ≥30 and eGFR <60). Mean systolic and diastolic blood pressure, mean glycosylated hemoglobin (HbA1c) level, and HbA1c variability (standard deviation [SD] of serial HbA1c values or HbA1c-SD) were calculated for the preceding 2 years. RESULTS: The prevalence of no DKD, non-severe DKD, and severe DKD was 37.2% (n=118), 37.0% (n=117), and 25.8% (n=82), respectively. HbA1c-SD and the triglyceride/high density lipoprotein cholesterol (TG/HDL-C) ratio correlated positively with uACR and negatively with eGFR. Multiple linear regression analyses showed that the HbA1c-SD and TG/HDL-C ratio were significantly related with eGFR. Multiple logistic regression analyses after adjusting for several risk factors showed that HbA1c-SD and the TG/HDL-C ratio were significant risk factors for severe DKD. CONCLUSION: The prevalence of DKD was about 60% in patients with T2DM and advanced DR. HbA1c variability and TG/HDL-C ratio may affect the development and progression of DKD in these patients.

Increased Level of Basophil CD203c Expression Predicts Severe Chronic Urticaria

Increased FcepsilonR1alpha expression with upregulated CD203c expression on peripheral basophils is seen in patients with chronic urticaria (CU). However, there has been no published report on the association between CD203c expression level and clinical disease activity in CU patients. To investigate whether the increase of basophil activation is associated with the disease activity of CU, we measured basophil CD203c expression using a tricolor flow cytometric method in 82 CU patients and 21 normal controls. The relationship between the percentage of CD203c-expressing basophils and clinical parameters was analyzed. The mean basophil CD203c expression was significantly higher in CU patients than in healthy controls (57.5% vs 11.6%, P or = 72% basophil CD203c expression and urticaria activity score (UAS)> or = 13 were significant predictors of severe CU (P = 0.005 and P = 0.032, respectively). These findings suggest that the quantification of basophil activation with CD203c at baseline may be used as a potential predictor of severe CU requiring another treatment option beyond antihistamines.

La Proteína C reactiva como predictor de la evolución de la neumopatía aguda comunitaria / Reactive Protein C as a predictor of the course of Community-Acquired Acute Pneumonia

La Proteína C reactiva es un biomarcador sensible, específico y buen predictor del fracaso terapéutico de las neumonías. A nivel mundial hay posturas controversiales sobre su utilidad. En nuestro medio no hay estudios reportados al respecto. El propósito de nuestro trabajo fue intentar predecir la evolución de la neumonía aguda comunitaria (NAC). A todos los pacientes con diagnostico de NAC ingresados al Hospital Español durante junio y julio de 2012 se les dosificó los niveles de PcR a las 24 y 72 hs. de iniciado el tratamiento. Se pudo correlacionar la buena evolución con la reducción de los niveles de PcR en un 25% (Δ PcR). También se observo que niveles elevados de PcR se correlacionan con la leucocitosis y el score CURB65. Se pudo concluir entonces que la PcR es un biomarcador de fácil acceso y buen predictor de la evolución de las NAC.

C-reactive protein is a biomarker sensitive, specific and good predictor of treatment failure in pneumonia. Worldwide there are controversial positions on their usefulness. In our country is not studies reported to respect. The purpose of our work was to try to predict the evolution of community acute pneumonia. All patients diagnosed with CAP admitted to Spanish Hospital during June and July 2012 were dosed RcP levels at 24 and 72 hours of starting treatment. We were able to correlate a good performance with reduced RcP levels by 25% (Δ RcP). We also observed that elevated RcP levels correlate with leukocytosis and CURB65 score. We conclude that RcP is a biomarker for easy access and good predictor of the development of the CAP.

Study on the expression of PINCH and vascular endothelial growth factor C in cervical squamous cell carcinoma / 中国综合临床

Objective To study the expression of PINCH and vascular endothelial growth factor C (VEGF-C) in cervical squamous cell carcinoma.Methods We detected the expression of PINCH and VEGF-C by immunohistochemistry SP in 58 cervical squamous cell carcinoma cases and 30 normal cervical epithelial tissue and analyzed their relationship to the clinical pathological features.Results The expression of PINCH and VEGF-C in 58 cervical squamous cell carcinoma(62.1％,36/58 ;67.2％,39/58 ) were higher than that in normal cervical epithelial tissue(0,0/30).The difference was significant( x2 =31.512,12.534,P ＜ 0.001 ).The expression of PINCH protein was not significantly associated with the age,tumor size and tumor differentiation grade ( P ＞ 0.05 ),but was associated with the lymph node metastasis and clinical stage ( x2 =9.090,8.236,P ＜ 0.001 ).The expression of VEGF-C had no significant correlationship with the age and tumor size( P ＞ 0.05 ) but had a correlationship with the lymph node metastasis,tumor differentiation grade and clinical stage( x2 =10.775,13.496,5.001,P ＜ 0.05 ).The expression of VEGF-C was correlated with the expression of PINCH protein( C =0.341,P ＜ 0.01 ).Conclusion It is possible that VEGF-C and PINCH take part in the development and progress of cervical squamous cell carcinoma and play an important role in the invasion and metastasis mechanism altogether.

Evaluación de un método de aglutinación con látex para la detección de proteína C reactiva / Agglutination evaluation method related to use of latex to detection of reactive C protein

Se evaluó un método de aglutinación con látex para la detección de proteína C reactiva (PCR) (PCR-Látex, CENTIS) empleándose como referencia un test turbidimétrico (PCR-Turbilátex, Spinreact). Se procesaron en paralelo 97 sueros de pacientes con clínica sugestiva de inflamación y sepsis. El análisis estadístico incluyó la determinación de la sensibilidad, especificidad, valor predictivo positivo (VPP), valor predictivo negativo (VPN) y límites de detección. Los resultados obtenidos de sensibilidad: 97,8 %; especificidad: 98,0 %; VPP: 97,8 %; VPN: 98,0 %, con un límite de detección de 6 mg/L, pueden considerarse de satisfactorios, lo que permite contar con un procedimiento sencillo, rápido y eficiente que no requiere de un equipamiento especial.

Agglutination evaluation method related to use of latex for detection of reactive C protein (RCP) (RCP-Latex, CENTIS) was evaluated, using as reference a turbidimetry test (RCP-Turbilatex, Spinreact). In parallel, 97 sera from patients with possibilities of inflammation and sepsis were processed. Statistical analysis included determination of sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and detection limits. Results achieved for sensitivity: 97,8 %; specificity: 98,0 %; NVP: 97,8 %; NPV: 98 % with a detection limit of 6 mg/L, may be considered of satisfactory, allowing to have a simple, fast, and efficient procedure without a especial equipment.

A Nationwide Survey about the Current Status of Glycemic Control and Complications in Diabetic Patients in 2006: The Committee of the Korean Diabetes Association on the Epidemiology of Diabetes Mellitus / 당뇨병

BACKGROUND: The Committee of the Korean Diabetes Association on the Epidemiology of Diabetes Mellitus performed a nationwide survey about the current status of glycemic control and diabetic complications in 2006. METHODS: The current study included 5,652 diabetic patients recruited from the rosters of endocrinology clinics of 13 tertiary hospitals in Korea. Age, gender, height, weight, waist circumference and blood pressure were investigated by standard method. Fasting and postprandial 2 hour glucose, glycosylated hemoglobin (HbA1c), lipid profiles, fasting insulin and c-peptide levels were measured. Microvascular (microalbuminuria, retinopathy and neuropathy) and macrovascular (coronary artery disease [CAD], cerebrovascular disease [CVD] and peripheral artery disease [PAD]) complications were reviewed in their medical records. RESULTS: Mean age of total subjects was 58.7 (+/- 11.6) years and duration of diabetes was 8.8 (0~50) years. Mean fasting and postprandial 2 hour glucose levels were 145.9 +/- 55.0 and 208.0 +/- 84.4 mg/dL, respectively. Their mean HbA1c was 7.9 +/- 1.9%: the percentage of patients within target goal of glycemic control (< 7% of HbA1c) was 36.7%. In this study, 30.3%, 38.3% and 44.6% of patients was found to have microalbuminuria, retinopathy and nephropathy, respectively. Prevalence of CAD, CVD and PAD was 8.7%, 6.7% and 3.0%, respectively. Diabetic complications were closely related with age, duration of diabetes and glycemic control, and this relationship was stronger in microvascular complications than macrovascular ones. CONCLUSION: Only about one third of patients with diabetes was found to reach target glycemic control in tertiary hospitals of Korea. More tight control is needed to reduce deleterious complications of diabetes in Korea.

B cell activation factor (BAFF) is a novel adipokine that links obesity and inflammation

B cell activation factor (BAFF) is a novel member of the TNF ligand superfamily, mainly produced by myeloid cells. BAFF has been shown to participate in B-cell survival and B- and T-cell maturation. BAFF expression in adipocytes has been recently demonstrated. In the current study, we verified that BAFF expression is increased during adipocyte differentiation. BAFF expression was augmented by TNF-alpha treatment and was decreased by rosiglitazone treatment. BAFF secretion in lean and in ob/ob mice sera were compared and smaller amount of BAFF was secreted in ob/ob mice. mRNA and protein expression were different between epididymal and visceral adipose tissue. BAFF expression was also increased in ob/ob mouse adipose tissue. We sought to identify known BAFF receptors (BAFF-R, BCMA, and TACI) in adipocytes, and determined that all three were present and upregulated during adipocyte differentiation. However, the expression of TACI was distinct from that of BAFF-R and BCMA under TNF-alpha and BAFF ligand treatment. BAFF-R and BCMA expression levels were upregulated under pro-inflammatory conditions, but TACI was reduced. Conversely, BAFF-R and BCMA expression levels were downregulated by rosiglitazone treatment, but TACI was increased. Taken together, our results suggest that BAFF may be a new adipokine, representing a link between obesity and inflammation.

The Relationship between Depression, Self-Care Activity and HbA1c in Clients with Type-2 Diabetes Mellitus / 기본간호학회지

PURPOSE: This study was done to investigate the relationship between depression, self-care activity and HbA1c in clients with type-2 diabetes mellitus (DM) clients. METHOD: This was a cross sectional descriptive study and data were collected between October 4 and 27, 2006 using a questionnaire. The participants were inpatients and outpatients with type-2 diabetes being seen in one of two hospitals. The questionnaire included general as well as disease related characteristics, a depression scale (CES-D), and the self-care activity scale developed by Kim (1996). HbA1c was obtained from medical records. RESULTS: There were 50 men (41.7%) and 70 women (58.3%). The mean scores for depression and self-care activity were 18.22+/-10.03 and 3.51, respectively. Almost half of the participants (46.7%) had an HbA1c greater than 7%. Self-care activity was negatively correlated with depression (r=-.436, p=0.00) and HbA1c (r=-.269, p=0.03). Depression was positively correlated with HbA1c (r=-.325, p=0.00). Participants who had diabetes education showed a significantly lower level of HbA1c than the group who had never had diabetes education. Conclusions: It is important to make ensure that patients with diabetes have education on diabetes mellitus and to screen and manage depression as part of the care for these patients.