Time Reverse Transcription-polymerase Chain Reaction Research Articles

Two metal-coordination polymers: Photocatalytic hydrogen production and clinical nursing value on gastric cancer combined with hyperthermic intraperitoneal chemotherapy by inducing induce oxidative stress response.

Two new coordination polymers (CPs) of {[Dy2(abtc)1.5(H2O)3(DMA)]·H2O·DMA}n (1; DMA=N,N-dimethylacetamide) and [NH2(CH3)2][Ni1.5(abtc)]·H2O (2) were synthesized through utilizing 3,3',5,5'-azobenzenetetracarboxylic acid (H4abtc) as an organic connector with the solvothermal reaction conditions. The HOMO-LUMO gap for ultra-stable complex 1 is 2.16eV through the detection of the ultraviolet-visible spectrum with an absorption edge of 570nm. The complex 1 reveals high catalytic effect for the ultraviolet-visible photocatalytic production of hydrogen, which is due to its light collection of the ligand with dye-like and porous architecture. Furthermore, the promotion effect of compounds 1 and 2 against the gastric cancer treatment was assessed when combined with intraperitoneal hyperthermia. Firstly, the inhibition of compounds 1 and 2 against the invasion and migration of cancer cells was evaluated with transwell assay. Moreover, the genes relative expression levels related with the reactive oxygen species (ROS) production in the cancer cells were also measured with real time reverse transcription-polymerase chain reaction (RT-PCR).

Transcriptional analyses of the effects of Catharanthus roseus L. medicinal plant extracts on some markers related to obesity and inflammation in 3T3-L1 mouse cell lines

Catharanthus roseus L. (C. roseus) is one of the medicinal plants used to treat diabetes. In this study, 3T3-L1 preadipocyte cell lines which are fully differentiated into adipocytes were utilized and these were treated with extracts derived from above ground part of C. roseus. The effect of these extracts on obesity and inflammation markers in cells was examined at the transcriptional level. Adipocyte lipid contents were measured by Oil Red O staining. Analyses, including changes related to adiposity and inflammation, were evaluated by measuring the relative mRNA expression levels of the genes of interest by the Real Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. The appropriate dose and durations for C. roseus were determined to be 12.5 μg/mL and 24- and 48 h respectively. The expression of the inflammation marker Interleukin-6 (Il-6) was decreased when C. roseus extract was administered to fully differentiated 3T3-L1 cells according to the determined dose and durations. Lipoprotein lipase (Lpl) and Fatty acid synthase (Fasn) gene expressions in fully differentiated cells decreased compared to the control after 24 h however, this effect was not observed after 48 h. Collagen V, has also been shown to be affected by treatment of fully differentiated 3T3-L1 cells with plant extracts in both 24- and 48-h periods.

RETRACTED ARTICLE: Two Mixed-Ligand Co(II)-Based Coordination Polymers: Treatment Activity on Pediatric Asthma Via Inducing Eosinophil Apoptosis Through Down-Regulating Bcl-2 Expression

In this work, two coordination polymers containing Co(II) ions as nodes, namely, [Co(NDC)(L)(H2O)]n (1, H2NDC = 1,4-naphthalene dicarboxylic acid) and [Co(DCTP)(L)·(H2O)2]n (2, H2DCTP = 2,5-dichloroterephthalic acid) were prepared based on 1,4-bis(5,6-dimethylbenzimidazole)butane (L), the flexible N-donor ligand and different dicarboxylate ligand. Their treatment activity on the pediatric asthma was evaluated in the biological research. Firstly, the enzyme linked immunosorbent assay (ELISA) detection was carried out to determine the IL-1β and TNF-α content in the bronchus. The inflammatory response in the bronchial epithelial cells was determined by the real time reverse transcription-polymerase chain reaction (RT-PCR). After that, the eosinophil apoptosis in the airway was measured. Finally, the western was performed for the confirmation of eosinophil apoptosis by detecting Bcl-2 and caspase-3.

Identification of serum exosomal miR-148a as a novel prognostic biomarker for breast cancer.

Breast cancer (BC) is one of the most common malignanciesD. Li, J. Wang, L.-J. Ma, H.-B. Yang, J.-F. Jing, M.-M. Jia, X.-J. Zhang, F. Guo, J.-N. Gao affecting females. Aberrant expression of microRNAs (miRNAs) has been associated with carcinogenesis of BC. The aim of our study was to investigate the correlation of serum exosomal miR-148a expression and the clinical outcome of patients with BC. Quantitative Real Time-Reverse Transcription Polymerase Chain Reaction (qRT-PCR) was applied to evaluate the expression level of serum exosomal miR-148a in patients with BC, patients with benign breast tumors, and healthy controls. Then, the clinical value of serum exosomal miR-148a in BC was evaluated. Serum exosomal miR-148a levels were gradually downregulated from healthy controls patients with benign breast tumors to BC patients. Serum exosomal miR-148a could well distinguish BC patients from healthy volunteers. The expression level of serum exosomal miR-148a in BC patients was significantly upregulated following surgery, while dropped in the cases with disease relapse. A significant association was found between serum exosomal miR-148a levels and the tumor-node-metastasis (TNM) stage, differentiation, and lymph node metastasis in BC. In addition, BC patients with lower expression of serum exosomal miR-148a levels suffered worse overall survival and disease-free survival than those with higher expression of serum exosomal miR-148a levels. Furthermore, serum exosomal miR-148a was an independent risk factor for BC. Our data have demonstrated that serum exosomal miR-148a is significantly reduced in patients with BC and downregulation of serum exosomal miR-148a is closely associated with unfavorable clinical outcome of BC, indicating that serum exosomal miR-148a might serve as a promising diagnostic and prognostic biomarker for BC.

A Deep Learning System to Screen Novel Coronavirus Disease 2019 Pneumonia

The real-time reverse transcription-polymerase chain reaction (RT-PCR) detection of viral RNA from sputum or nasopharyngeal swab had a relatively low positive rate in the early stage of coronavirus disease 2019 (COVID-19). Meanwhile, the manifestations of COVID-19 as seen through computed tomography (CT) imaging show individual characteristics that differ from those of other types of viral pneumonia such as influenza-A viral pneumonia (IAVP). This study aimed to establish an early screening model to distinguish COVID-19 from IAVP and healthy cases through pulmonary CT images using deep learning techniques. A total of 618 CT samples were collected: 219 samples from 110 patients with COVID-19 (mean age 50 years; 63 (57.3%) male patients); 224 samples from 224 patients with IAVP (mean age 61 years; 156 (69.6%) male patients); and 175 samples from 175 healthy cases (mean age 39 years; 97 (55.4%) male patients). All CT samples were contributed from three COVID-19-designated hospitals in Zhejiang Province, China. First, the candidate infection regions were segmented out from the pulmonary CT image set using a 3D deep learning model. These separated images were then categorized into the COVID-19, IAVP, and irrelevant to infection (ITI) groups, together with the corresponding confidence scores, using a location-attention classification model. Finally, the infection type and overall confidence score for each CT case were calculated using the Noisy-OR Bayesian function. The experimental result of the benchmark dataset showed that the overall accuracy rate was 86.7% in terms of all the CT cases taken together. The deep learning models established in this study were effective for the early screening of COVID-19 patients and were demonstrated to be a promising supplementary diagnostic method for frontline clinical doctors.

Shuxuening injection facilitates neurofunctional recovery via down-regulation of G-CSF-mediated granulocyte adhesion and diapedesis pathway in a subacute stroke mouse model

Post-stroke neural damage is a serious health concern which does not yet have an effective treatment. We have shown previously that Shuxuening injection (SXNI), a Ginkgo biloba extract-based natural medicine, protects brain after an acute ischemic stroke, but its efficacy for post-stroke recovery is not known. This study was to investigate whether SXNI can improve the prognosis of stroke at a subacute phase. Mice with cerebral ischemia-reperfusion injury (CIRI) were established by middle cerebral artery occlusion (MCAO), and drugs or saline were injected by the tail vein every 12 h after reperfusion. The therapeutic effect of SXNI was evaluated by survival rate, modified neurologic severity scores (mNSS), open-field test, locomotive gait patterns, cerebral infarction volume, brain edema and histopathological changes. Subsequently, a combined method of RNA-seq and Ingenuity® Pathway Analysis (IPA) was performed to identify key targets and pathways of SXNI facilitating the prognosis of stroke in mouse brain. The results of the transcriptome analysis were verified by real time reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), western blot (WB) and immunohistochemistry (IHC). The experimental results showed that in the new subacute stroke model, SXNI markedly improves the survival rate, neurological and motor functions and histopathological changes, and significantly reduces cerebral infarction and edema volume. RNA-seq analysis of subacute stroke mice with or without SXNI (3 mL/kg) indicated 963 differentially expressed genes (DEGs) with a fold change ≥ 1.5 and a P-value ≤ 0.01. IPA analysis of DEGs showed that granulocyte adhesion and diapedesis ranked first in the pathway ranking, and the most critical gene regulated by SXNI was G-csf. Simultaneously, RT-PCR, ELISA, WB and IHC results demonstrated that SXNI not only obviously reduced the mRNA expression levels of key genes G-csf, Sele and Mac-1 in this pathway, but also significantly decreased the protein expression levels of G-CSF in serum and E-selectin and MAC-1 in brain tissues. In summary, our research suggested that SXNI can exert a remarkable neurofunctional therapeutic effect on stroke mice via down-regulating G-CSF to inhibit granulocyte adhesion and diapedesis. This study provides experimental evidence that SXNI may fulfill the need for stroke medicine targeting specifically at the recovery stage.

Epidemiological and initial clinical characteristics of patients with family aggregation of COVID-19

BackgroundSince December 2019, a new outbreak of the coronavirus disease 2019 (COVID-19) in Wuhan (Hubei, China) and rapidly spread throughout China, however, confirmed cases are still increasing worldwide. ObjectivesTo investigate the epidemiological history and initial clinical characteristics of 10 patients with family aggregation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Western Chongqing, China. Study designTen patients positive for SARS-CoV-2 nucleic acid detection by real time Reverse Transcription-Polymerase Chain Reaction (RT-PCR), were collected from The People's Hospital of Dazu District, Chongqing. Epidemiological data and laboratory and imaging results were collected on the first day of admission, and analyzed based on the Diagnosis and Treatment Guideline for COVID-19 (5th edition, China). ResultsOf the 10 cases, case A had a history of a temporary stay in Wuhan and transmitted the virus to the others through family gathering, living together, and sharing vehicles. The average age was 56.5 years (± 11.16), six patients were males, and the incubation period was 2–14 days. Dry cough was the main symptom, followed by fever and fatigue. Most patients were clinically classified as ordinary-type, with three cases being severe-type. Chest computed tomography results were nonspecific, mainly with ground-glass attenuation and/or shadow images. Extensive lesion distribution was seen in severe cases. CD4+ lymphocyte counts were 61, 180, and 348 cells/uL in severe-type patients, respectively. Notably, viral nucleic acid values in nasopharyngeal swabs were lower (19, 25, and 26) than those of ordinary-type patients, suggesting a higher viral load. Neutrophil-lymphocyte ratio (NLR) was also higher in severe-type patients ConclusionsInitial examination results of lower CD4+ lymphocyte counts and RT-PCR-CT values coupled with higher NLR may indicate the severity of COVID-19 infection for these family clusters.

Serum CEA and CYFRA Levels in ALK-rearranged NSCLC Patients: Correlation With Distant Metastasis.

To clarify the correlation between serum levels of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA) and metastasis and survival in anaplastic lymphoma kinase (ALK)-rearranged non-small cell lung cancer (NSCLC) patients. CEA and CYFRA levels in 131 ALK-rearranged NSCLC patients were determined using fluorescence in situ hybridization (FISH), real time-reverse transcription polymerase chain reaction, and immunohistochemistry, using biopsy specimens, cytology specimens, and plasma specimens. Cut-off value of each marker was determined as 10 ng/ml. In logistic regression analysis, higher levels of both markers had a positive relationship with bone metastases, and higher levels of CYFRA was relevant to liver metastases, and multiple-organ metastases. However, these markers were not proven to be poor prognostic factors in Cox's proportional model analysis. Elevated serum CEA and CYFRA levels seem to provide useful clinical information about presence of bone and liver metastasis and multiple-organ metastases, although they were not a powerful indicator of prognosis. These two markers may suggest the extension of metastasis and would be helpful in considering treatment options.

Laboratory surveillance of chikungunya in Madhya Pradesh, India (2016-2017).

Background & objectives:Chikungunya (CHIK) is a neglected, re-emerging arboviral disease. Limited information on CHIK-confirmed cases during interepidemic period is available from India. This surveillance study was conducted in Madhya Pradesh (MP), India, during the years 2016-2017, to provide information about CHIK cases.Methods:Blood samples collected from patients suspected having CHIK were tested by immunoglobulin (Ig) IgM ELISA or real time reverse transcription-polymerase chain reaction (rRT-PCR) for the detection of CHIK virus (CHIKV)-specific IgM antibodies or viral RNA, respectively. Partial envelope-1 gene sequencing was done. Clinical and demographic data were collected and analyzed.Results:Of the 4019 samples tested, 494 (12.2%) were found positive for CHIKV infection. The positivity was detected in both rural and urban areas. The mean age of CHIK-positive cases was 33.12±18.25 yr. Headache and joint pain were the most prominent symptoms, 34.6 per cent (171/494) of the CHIK cases required hospitalization and six patients with CHIKV infection died. The East/Central/South African genotype of CHIKV was found to be circulating in the study area.Interpretation & conclusions:Our study recorded a higher CHIK positivity during 2016-2017 in comparison to earlier reports from MP, India. A high proportion of CHIK cases required hospitalization and deaths were also reported, which indicated the severity of the disease in the study area. In-depth molecular analysis of the virus and other risk factors is essential to understand the trends in disease severity.

MiRNA-212 acts as a tumor-suppressor in colorectal carcinoma through targeting SOX4.

Colorectal cancer is a common gastrointestinal cancer, with mortality ranking the third all over the world. MicroRNA-212 (miR-212), located on chromosome 17p13.3, was lowly expressed in a variety of tumors. The purpose of our study was to explore the effects of miR-212 on colorectal cancer (CRC) cells. Quantitative Real Time-Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blot were employed to evaluate the levels of mRNAs and proteins. The transwell assay was performed to calculate the abilities of migration and invasion. The Luciferase reporter assay was utilized to verify miR-212 targeting to the 3'-UTR of SOX4 mRNA. Statistical analysis was applied to analyze the experimental data. MiR-212 was lowly expressed in CRC tissues and cell lines, while the expression of SRY-box 4 (SOX4) was overexpressed. Exogenous increasing of miR-212 or knockdown of SOX4 inhibited the migration, invasion and epithelial-mesenchymal transition (EMT) in CRC cells. Moreover, the expression of miR-212 had a negative connection with SOX4 expression, and miR-212 mediated the expression of SOX4 by directly binding to the 3'-UTR of SOX4 mRNA. In addition, low expression of miR-212 or overexpression of SOX4 was associated with poor prognosis of colorectal cancer patients. MiR-212 inhibited the migration, invasion and EMT by direct targeting to the 3'-UTR of SOX4 mRNA in colorectal cancer. The newly identified miR-212/SOX4 axis provides novel insight for colorectal treatment.

RETRACTED ARTICLE: A New Cluster-Based MOF for Selective Gas Sorption and Treatment Effect on Acute glomerulonephritis by Reducing NF-κb Pathway Activation and Cytokines Release

A novel Co(II)-containing metal–organic framework [Co3(OH)2(H2TCPP)2](DMF)3 (1) based on a linear trinuclear Co(II) cluster-based unit has been successfully prepared by using a nanosized tetracarboxylate ligand 2,3,5,6-tetrakis(4-carboxyphenyl)pyrazine (H4TCPP) as the organic building block under the solvothermal reaction. Gas adsorption studies reveal that the mixtures of C2H2/CH4 and C2H2/CO2 could be selectively separated by the obtained resulting solvent free 1 (denoted as 1a hereafter). To deep evaluate the protective function of complex for acute glomerulonephritis (AGN) disease in vitro, the following experiments were conducted. (a) Real time Reverse transcription-Polymerase Chain Reaction (RT-PCR) was carried out to determine the relative expression of Nuclear factor kappa beta (nf-κb) and Tumor Necrosis Factor-α (tnf-α) in the glomerulus cells after bacterial infection along with complex 1 treatment. (b) The enzyme linked immunosorbent assay (ELISA) was conducted to determine the IL-1β and IL-6 released by glomerular cells after complex 1 treatment. (c) The results obtained through molecular docking exhibited possible regulation mechanism on the binding affinity between compound 1 and target protein.

MiR-30a-3p promotes ovariectomy-induced osteoporosis in rats via targeting SFRP1.

To explore the effect of miR-30a-3p on osteoporosis in rats after ovariectomy. The ovariectomized (OVX) rat model was established to mimic postmenopausal osteoporosis. The primary bone marrow mesenchymal stem cells (BMMSCs) isolated from female rats were transfected with mimic or inhibitor. Real Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting were used to examine the expressions of miR-30a-3p and osteoporosis-related proteins. The bone mineral density (BMD) was detected via micro-Computed Tomography (CT) and the bone histomorphometry was performed. Luciferase assay was also performed to confirm whether SFRP1 is a target of miR-30a-3p. According to micro CT, BMD significantly declined in OVX group. MiR-30a-3p and SFRP1 were negatively correlated after ovariectomy. SFRP1 was acknowledged as a target of miR-30a-3p. Besides, the miR-30a-3p inhibitor promoted osteogenic differentiation in vitro and bone formation in vivo. MiR-30a-3p inhibitor promotes bone formation through decreasing SFRP1 expression and miR-30a-3p may be a potential novel molecular target in the treatment of osteoporosis.

Sea bass (Lateolabrax maculatus) accelerates wound healing: A transition from inflammation to proliferation

Ethnopharmacological relevanceSea bass (Lateolabrax maculatus) has been used for dietary therapy practice for wound healing of puerperal or surgery patients in China. Traditional Chinese medicinal books also documented that sea bass can be used to manage inflammation-associated conditions such as wound, miscarriage and cough. Some studies also proved that dietary supplement with fish benefited for treating many inflammatory - associated conditions, such as cardiovascular disease, ulcerative colitis and hyperlipidemia. However, the studies on the pharmacological mechanisms of wound healing efficacy of sea bass remain lack of investigation. Aim of the studyThe aim of this study is to investigate the molecular mechanisms of sea bass on wound healing efficacy. Establishing a further justification for clinical application of aqueous extract of sea bass (ASB) in treating wound healing. Materials and methodsTransition from inflammation to proliferation phase treated as the critical step in wound repair which were investigated via in vitro and in vivo study. A series of inflammatory mediators associated with wound healing and proliferation effects of fibroblasts upon treatments were studied via Western blotting, enzyme-linked immunosorbent assay (ELISA), real time reverse transcription-polymerase chain reaction (RT-PCR) and scratch assay. The cutaneous wound model was applied on skin wound healing study to observe the healing process in C57BL/6 mice upon ASB treatments. Hematological parameters and tumor necrosis factor-α (TNF-α) secretions in serum were determined. Histopathological examinations were conducted by hematoxylin and eosin (H&E) staining and Masson staining. Immunofluorescence were performed to identify infiltrating neutrophils (MPO) and α-smooth muscle actin (α-SMA). ResultsResults showed that ASB significantly reduced the production of inflammatory mediators cyclooxygenase-2 (COX-2), nitrite oxide (NO) production and TNF-α. The phosphorylation and nuclear protein levels of transcription factor nuclear factor-κB (NF-ĸB) in toll-like receptor 4 (TLR4) signaling were decreased by ASB treatment as well. Wound closure rate and cyclin D1 expression level of fibroblasts were significantly increased by ASB treatments. Moreover, cutaneous wound model in C57BL/6 mice presented many similarities in appearance to the process of wound healing. ConclusionsThe in vitro study demonstrated an inhibitory effect of ASB on the inflammatory mediators regulated by TLR4 signaling pathways, providing evidence that ASB treatment potentially accelerate the wound healing through migration and proliferation enhancement. Additionally, the in vivo study suggested that ASB treatment has a potential in accelerating the proliferation phase of wound healing via well-organized abundant collagen deposition, angiogenesis and re-epithelialization in wounds. The present findings can be treated as a pharmacological basis for the folk use of sea bass and further studies in biological and medical fields.

An in silico analysis of human sperm genes associated with asthenozoospermia and its implication in male infertility.

Asthenozoospermia is the most common clinical symptom of male infertility. Molecular markers associated with asthenozoospermia spermatozoa are scarcely identified. The objective of this study was to screen the differentially expressed genes (DEGs) in asthenozoospermia spermatozoa and assess the underlying bioinformatics roles in regulation of sperm quality.Based on gene expression omnibus (GEO) database, the GSE22331, GSE1133, and GSE4193 expression profile data were downloaded. The DEGs of asthenozoospermia spermatozoa were identified. Germ cell specific genes in DEGs were further screened. Then, gene ontology (GO) and over-representation analysis of DEGs were performed, followed by protein-protein interaction (PPI) network analysis. Expressions of selected genes of TEX11, ADAMTS5, ASRGL1, GMCL1, PGK2, KLHL10 in normozoospermia and asthenozoospermia spermatozoa were identified using real time Reverse Transcription-Polymerase Chain Reaction (RT-PCR).A total of 1323 DEGs were identified, including 1140 down-regulated genes. Twenty one and 96 down-regulated genes were especially expressed in spermatogonia and round spermatids, suggesting their testicular origins and influences on sperm quality. Bioinformatics analysis showed enriched functions of ubiquitin-like protein transferase or protein binding activities in down-regulated genes. Expressions of selected genes were validated by RT-PCR, which was consistent with bioinformatical results.The present study provided a novel insight into the understanding of sperm quality, and a potential method and dataset for the diagnosis and assessment of sperm quality in the event of male infertility.

Developing a multiplex real-time RT-PCR for simultaneous detection of Pepper chat fruit viroid and Columnea latent viroid

Tomato (Solanum lycopersicum L.) is one of the most important vegetables in the world and Asian countries are the center for hybrid tomato seed production. Seed-borne diseases, if not managed well through effective seed health assay, could seriously hamper the global seed trade. Pepper chat fruit viroid (PCFVd) and Columnea latent viroid (CLVd) are two emerging and economically important pospiviroid species infecting tomato and other solanaceous crops. With their greater sequence diversity from those of other common tomato-infecting pospiviroids, it is necessary to develop a new detection method that could effectively detect PCFVd and CLVd, useful for seed health assay. In the present study, we developed a multiplex real time (quantitative) reverse transcription-polymerase chain reaction (qRT-PCR) to detect PCFVd and CLVd in a single tube, thus to save time and money. Using known viroid-contaminated seeds, this multiplex qRT-PCR could detect CLVd and PCFVd from a single contaminated seed, suggesting its sensitivity for seed health assay. The effectiveness of this newly developed multiplex qRT-PCR for detection of PCFVd and CLVd was validated using field-produced tomato plant tissue or commercial tomato seeds.

Epidemiology of circulating human influenza viruses from the Democratic Republic of Congo, 2015.

IntroductionThe establishment of the influenza sentinel surveillance system in Kinshasa, Bas Congo, Maniema, Katanga, and Kasai Provinces allowed generation of important data on the molecular epidemiology of human influenza viruses circulating in the Democratic Republic of Congo (DRC). However, some challenges still exist, including the need for extending the influenza surveillance to more provinces. This study describes the pattern of influenza virus circulating in DRC during 2015.MethodologyNasopharyngeal swabs were collected from January to December 2015 from outpatients with influenza-like illness (ILI) and in all hospitalized patients with Severe Acute Respiratory Infection (SARI). Molecular analysis was done to determine influenza type and subtype at the National Reference Laboratory (NRL) in Kinshasa using real time reverse transcription-polymerase chain reaction (rRT-PCR). Analysis of antiviral resistance by enzyme inhibition assay and nucleotide sequencing was performed by the Collaborating center in the USA (CDC, Atlanta).ResultsOut of 2,376 nasopharyngeal swabs collected from patients, 218 (9.1%) were positive for influenza virus. Among the positive samples, 149 were characterized as influenza virus type A (Flu A), 67 as type B (Flu B) and 2 mixed infections (Flu A and B). Flu A subtypes detected were H3N2 and H1N1. The Yamagata strain of Flu B was detected among patients in the country. Individuals aged between 5 and 14 years accounted for the largest age group affected by influenza virus. All influenza viruses detected were found to be sensitive to antiviral drugs such as oseltamivar, zanamivir, peramivir and laninamivar.ConclusionThe present study documented the possible involvement of both circulation of Flu A and B viruses in human respiratory infection in certain DRC provinces during 2015. This study emphasises the need to extend the influenza surveillance to other provinces for a better understanding of the epidemiology of influenza in DRC. It is envisioned that such a system would lead to improved disease control and patient management.

Burden of dengue among febrile patients at the time of chikungunya introduction in Piedecuesta, Colombia.

ObjectiveTo estimate the age‐specific incidence of symptomatic dengue and chikungunya in Colombia.MethodA passive facility‐based fever surveillance study was conducted among individuals with undifferentiated fever. Confirmatory diagnostics included serological and molecular tests in paired samples, and surveillance's underreporting was assessed using capture–recapture methods.ResultsOf 839 febrile participants 686 completed the study. There were 33.2% (295/839) dengue infections (51% primary infections), and 35.9% (191/532) of negative dengue cases there were chikungunya cases. On average, dengue cases were younger (median = 18 years) than chikungunya cases (median = 25 years). Thrombocytopaenia and abdominal pain were the main dengue predictors, while presence of rash was the main predictor for chikungunya diagnosis. Underreporting of dengue was 31%; the estimated expansion factors indicate an underreporting rate of dengue cases of threefold for all cases and of almost sixfold for inpatients.ConclusionsThese findings highlight the ongoing coexistence of both arboviruses, a distinct clinical profile of each condition in the study area that could be used by clinicians to generate a differential diagnosis, and the presence of underreporting, mostly among hospitalised cases.

Expression signature of lysosomal-associated transmembrane protein 4B in hepatitis C virus-induced hepatocellular carcinoma.

Hepatocellular carcinoma is a lethal disease worldwide and therefore the establishment of novel diagnostic biomarkers is imperative. In this study, it was hypothesized that an abnormal expression of the lysosomal-associated protein transmembrane 4 beta ( LAPTM4B) gene is crucial in the pathogenesis of hepatitis C virus-mediated hepatocellular carcinoma; hence we investigated the expression profile of LAPTM4B in hepatitis C virus-induced hepatocellular carcinoma. A group of 189 consecutive patients (hepatitis C virus-related hepatocellular carcinoma as tumor cases; n=93, hepatitis C virus-related cirrhotics as disease controls; n=96) opting for living donor liver transplantation as a therapeutic surgical regimen were recruited with informed consent. Additionally, paired adjacent non-tumorous tissues (n=93) obtained from cases were also included. Serum LAPTM4B protein concentrations were assessed by third-generation enzyme-linked immunosorbent assay and LAPTM4B mRNA, and protein expressions at tissue level were determined by quantitative real time reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry techniques, respectively. LAPTM4B protein concentrations in sera of patients were higher ( p<0.001) in tumor cases (1.25±0.25 ng/ml) compared to disease controls (0.53±0.28 ng/ml). Our study also depicts positive clinicopathological correlations between alpha-fetoprotein titers (b=0.65; p<0.001), quantitative hepatitis C virus RNA copies (b=0.33; p<0.001), and LAPTM4B protein concentrations, all in sera of patients. In addition, qRT-PCR and immunohistochemistry analyses revealed a significantly higher ( p<0.05) tissue LAPTM4B mRNA and protein expression, respectively, in tumor cases rather than in non-tumorous tissues and disease controls. Together, our results illustrate the LAPTM4B gene as a diagnostic biomarker in patients with hepatocellular carcinoma having documented evidence of chronic hepatitis C virus infection.

Development of a sensitive real-time PCR detection method for citrus tatter leaf virus

Citrus tatter leaf virus (CTLV) is a graft-transmissible capillovirus that causes the tatter leaf disease of citrus and bud union incompatibility between scion and trifoliate hybrid rootstocks which can result in serious economic losses. Although several CTLV detection protocols have been described, there is a need for a highly sensitive protocol that can be used in routine diagnostic tests for virus-free budwood certification programs. To address this need, a TaqMan® chemistry based real time reverse transcription-polymerase chain reaction (RT-qPCR) assay was developed to detect CTLV from citrus plants. The nucleotide sequence analysis based on full genomic sequences of eight CTLV isolates available from GenBank showed that 3′ region of the viral genome is more conserved among different CTLV isolates than the rest of the viral genome. A new set of primers and a TaqMan® probe were designed from a conserved 500 nt-long fragment at the 3′ end of the CTLV genome for the detection of CTLV by RT-qPCR. The newly designed primers for CTLV RT-qPCR assay had ca. 98.6% amplification efficiency and showed 100% CTLV detection rate when they were tested for previously known CTLV-positive trees infected with different CTLV isolates. The sequencing of 132 bp-long RT-qPCR amplicon followed by BLASTn search showed that it had 94% to 98% sequence identity to those of various CTLV isolates available in GenBank, confirming the specificity of the assay to reliably detect CTLV.

Biomolecular analysis of matrix proteoglycans as biomarkers in non small cell lung cancer.

Matrix proteoglycans (PGs) have shown promise as biomarker in malignancies. We employed agarose gel eletrophoresis, quantitative real- time reverse transcription-polymerase chain reaction and immunohistochemistry to evaluate the content of sulfated glicosaminoglycans (chondroitin sulfate and heparan sulfate) and expression of PG (biglycan, glypican, perlecan, syndecan e versican) in patient-matched normal and tumor tissues obtained from resected specimens of lung cancer. A significant increase of heparan sulfate (HS) and chondroitin sulfate (CS) concentrations was found in tumor tissue samples when compared to normal lung tissue samples. HS was also significantly increased in adenocarcinomas compared to squamous cell carcinomas. PG gene expression, with exception of syndecan, were significantly decreased in tumor tissue compared to normal lung, coinciding with significant decrease of PG protein levels in tumor cells and stroma compared to normal lung tissue (Kappa coefficient 0.41, 0.42 and 0,28, respectively). Women patients (p = 0.02), non smokers (p = 0.05), T stage (p = 0.009), N stage (p = 0.03) and adenocarcinoma (p = 0.05) were associated with improved overall survival (OS). Patients presenting tumors with low concentration of sulfated GAG and high PGs levels presented better OS compared to patients with high concentration of sulfated GAG and low expression of PGs. Cox regression model controlled by gender, tobacco history and histological type, showed that patients with high perlecan and versican expression in tumor presented respectively high probability of life (β risk 11.64; 1.27 to 15.90) and low risk of death (β risk 0.11; 0.02-0.51). The combined approach suggest matrix (PGs) as biomarkers in lung cancer.