Correlation between CT based radiomics features and gene expression data in non-small cell lung cancer.

OBJECTIVE: Radiogenomics investigates radiographic imaging phenotypes associated with gene expression patterns. This study aims to explore relationships between CT imaging radiomics features and gene expression data in non-small cell lung cancer (NSCLC). METHODS: Eighty-nine NSCLC patients are included in the study. Radiomics features are extracted and selected to quantify the phenotype of tumors on CT-scans. Co-expressed genes are also clustered and the first principal component of the cluster is represented, which is defined as a metagene. Then, statistical analysis was performed to assess association of CT radiomics features with metagenes. In addition, predictive models are built and metagene enrichment are conducted to further evaluate performance of NSCLC radiogenomics statistically and biologically. RESULTS: There are 187 significant pairwise correlations between a CT radiomics feature and a metagene of NSCLC, where eighteen metagenes are annotated with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) terms. Metagenes are predicted in terms of radiomics features with an accuracy of 41.89% -89.93%. CONCLUSIONS: This study reveals the associations between CT imaging radiomics features and NSCLC co-expressed gene sets. The findings suggest that CT radiomics features can reflect important biological information of NSCLC patients, which may have a significant clinical impact as CT is routinely used in clinical practice, assisting in improving medical decision-support at low cost.

[Expression of the Ces5a gene in the rat testis].

OBJECTIVE: To study the expression of the Ces5a gene in the development of the rat testis. METHODS: Using RT-PCR, Western blot, immunohistochemistry and HE staining, we determined the mRNA transcription level, protein expression and localization of the Ces5a gene in the testes of three litters of rats at different postnatal (PN) days. RESULTS: The expression of Ces5a mRNA was found in the testis tissue of the rats at 2－65 PN days, low at 2－12 days, decreased to the lowest level at 14－16 days (P < 0.05), but significantly increased at 20－35 days (P < 0.05), and elevated to the highest level at 40－65 days (P < 0.05). The expression of the Ces5a protein was also observed in the testis tissue of the rats at 2－65 PN, low at 2－12 days, with no significant change at 14－16 days (P > 0.05), but markedly increased at 20－35 days (P < 0.05), and again with no significant change at 40－65 days (P > 0.05). The Ces5a protein was expressed in the spermatogonia, spermatocytes and round sperm cells. CONCLUSIONS: The Ces5a gene may be involved in the proliferation and meiosis of rat spermatogonia and play a special role in round spermatogenesis and sperm deformation.

[Airway inflammation induced by Poly(I:C) stimulation in the late stage of respiratory syncytial virus infection in mice and its mechanism].

OBJECTIVE: To investigate the pathogenic mechanisms of airway inflammation and recurrent wheezing induced by recurrent respiratory virus infection after respiratory syncytial virus (RSV) infection. METHODS: Sixty-four female BALB/c mice (aged 6-8 weeks) were randomly divided into four groups: control, RSV, Poly(I:C), and RSV+Poly(I:C) (n=16 each). The bronchoalveolar lavage fluid (BALF) was collected on the 3rd day after Poly(I:C) administration, and the total cell number and differential counts in BALF were determined. Hematoxylin-eosin staining was used to observe pulmonary pathological changes. The airway responsiveness was detected. ELISA was used to measure the levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-13 (IL-13), matrix metallopeptidase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in BALF. RESULTS: Compared with the other three groups, the RSV+Poly(I:C) group had significant increases in the total number of inflammatory infiltrating cells in the airway, airway responsiveness, and MMP-9 level in BALF (P<0.05). The RSV+Poly(I:C) group showed more severe pulmonary tissue injuries compared with the control and RSV groups (P<0.01). Compared with the RSV group, the RSV+Poly(I:C) group showed significant reductions in the levels of IL-4 and TIMP-1 in BALF (P<0.01). CONCLUSIONS: Viral re-infection in the late stage of RSV infection may cause an imbalance of MMP-9/TIMP-1 expression and thus contribute to aggravated airway inflammation.

Association of Pneumococcal Carriage and Expression of Foxp3+ Regulatory T Cells and Th17 Cells in the Adenoids of Children.

BACKGROUND: Pneumococcal carriage in the nasopharynx is a primary means of transmission and a necessary prerequisite for pneumococcal disease. OBJECTIVES: We analyzed the relationship between expressions of Foxp3+ regulatory T (Treg) cells and Th17 cells, and pneumococcal carriage in the adenoids of children who were either positive or negative for pneumococci. METHODS: We collected adenoidal tissue and nasopharyngeal swab samples from children undergoing an adenoidectomy. Adenoidal mononuclear cells were isolated, cultured and then stimulated with culture concentrated supernatant (CCS) obtained from a D39 bacterial strain. RESULTS: Foxp3+ Treg cells were upregulated and Th17 cells were downregulated in populations of adenoidal mononuclear cells obtained from the pneumococcus-positive group. Following CCS stimulation, the increment in Foxp3+ Treg cells in the pneumococcus-positive group was significantly greater than that in the pneumococcus-negative group, while the increment in Th17 cells was less as compared to that in the pneumococcus-negative group. These results were consistent with variations in levels of Foxp3 mRNA and retinoic acid receptor-related orphan receptor-γt mRNA in adenoidal mononuclear cells. Levels of IL-17A and IL-6 in adenoid tissue were higher in the pneumococcus-negative group, and the levels of TGF-ß in adenoid tissue were lower in the pneumococcus-negative group compared to the pneumococcus-positive group. Pneumococcal carriage in children was closely associated with the expressions of Foxp3+ Treg and Th17 cells in the adenoid. CONCLUSION: Upregulation of Foxp3+ Treg cells might downregulate the production of Th17 cells in the adenoid, resulting in decreased scavenging of Streptococcus pneumoniae and chronic pneumococcal carriage.

Effect of screening colonoscopy frequency on colorectal cancer mortality in patients with a family history of colorectal cancer.

BACKGROUND: Colorectal cancer is a common malignant tumor in China, and its incidence in the elderly is increasing annually. Inflammatory bowel disease is a group of chronic non-specific intestinal inflammatory diseases, including ulcerative colitis and Crohn's disease. AIM: To assess the effect of screening colonoscopy frequency on colorectal cancer mortality. METHODS: We included the clinicopathological and follow-up data of patients with colorectal cancer who underwent laparoscopic colectomy or open colectomy at our Gastrointestinal Department between January 2019 and December 2022. Surgical indicators, oncological indicators, and survival rates were compared between the groups. The results of 104 patients who met the above criteria were extracted from the database (laparoscopic colectomy group = 63, open colectomy group = 41), and there were no statistically significant differences in the baseline data or follow-up time between the two groups. RESULTS: Intraoperative blood loss, time to first ambulation, and time to first fluid intake were significantly lower in the laparoscopic colectomy group than in the open colectomy group. The differences in overall mortality, tumor-related mortality, and recurrence rates between the two groups were not statistically significant, and survival analysis showed that the differences in the cumulative overall survival, tumor-related survival, and cumulative recurrence-free rates between the two groups were not statistically significant. CONCLUSION: In elderly patients with colorectal cancer, laparoscopic colectomy has better short-term outcomes than open colectomy, and laparoscopic colectomy has superior long-term survival outcomes compared with open colectomy.

Protection against hyperoxia-induced lung fibrosis by KGF-induced MSCs mobilization in neonatal rats.

MSCs have been shown to improve functional and pathological outcome in lung fibrosis. However, low in vivo cell engraftment of the transplanted cells limits their overall effectiveness. KGF (also known as FGF-7) is a critical mediator of pulmonary epithelial repair through stimulation of epithelial cell proliferation. However, the role of KGF in MSCs and its therapeutic effects have not been identified. In this study, we investigated the effect of KGF on MSCs and its preventive role in hyperoxia-induced fibrosis in neonatal rats. Neonatal rats exposed to normoxia or hyperoxia were randomly assigned to receive intraperitoneal injections of normal saline (PL), MSCs, or KGF pretreated MSCs on the fourth day of exposure. Our results showed that as compared to PL, while MSCs attenuated lung fibrosis, KGF pretreated MSCs exhibited enhanced preventive effect against lung fibrosis. This effect was partly attributed to enhanced mobilization of MSCs to the fibrotic lungs. In addition, the SHH signaling pathway, which is associated with the differentiation of stem cells was activated by KGF. Our data suggest that MSCs, especially KGF preconditioned MSCs, can attenuate lung fibrosis and KGF may regulate the MSCs behavior by activating SHH pathway.

Kawasaki disease with peritonsillar abscess as the first symptom: A case report.

BACKGROUND: Kawasaki disease (KD), also known as mucocutaneous lymph node syndrome, is an acute, self-limiting vasculitis of unknown aetiology that mainly involves the medium and small arteries and can lead to serious cardiovascular complications, with a 25% incidence of coronary artery aneurysms. Periton-Sillar abscesses are a rare symptom of KD and is easily misdiagnosed at its early stages. CASE SUMMARY: A 5-year-old boy who presented to a community hospital with a 3-d fever, difficulty in opening his mouth, and neck pain and was originally treated for throat infection without improvement. On the basis of laboratory tests, ultrasound of submandibular and superficial lymph nodes and computed tomography of the neck, the clinician diagnosed the periamygdala abscess and sepsis that did not resolve after antibiotic therapy. On the fifth day of admission, the child developed conjunctival congestion, prune tongue, perianal congestion and desquamation, and slightly stiff and swollen bunions on both feet. A diagnosis of KD was reached with complete remission after intravenous immunoglobulin treatment. CONCLUSION: Children with neck pain, lymph node enlargement, or airway obstruction as the main manifestations are poorly treated with intravenous broad-spectrum antibiotics. Clinicians should not rush invasive operations such as neck puncture, incision, and drainage and should be alert for KD when it cannot be explained by deep neck space infection and early treatment with aspirin combined with gammaglobulin.

A comparison of the use of contrast media with different iodine concentrations for enhanced computed tomography.

Objective: In this study, we compared the enhancement of blood vessels and liver parenchyma on enhanced computed tomography (CT) of the upper abdomen with two concentrations of contrast media (400 and 300 mg I/mL) based on similar iodine delivery rate (IDR) of 0.88 and 0.9 g I/s and iodine load of 450 mg I/kg. Methods: We randomly assigned 160 patients into two groups: iomeprol 400 mg I/mL (A group) and iohexol 300 mg I/mL (B group). The CT attenuation values of the main anatomical structures in the two groups with different scanning phases were measured and the image quality of the two groups was analyzed and compared. The peak pressure and local discomfort (including fever and pain) during contrast medium injection were recorded. Results: The mean attenuation value of the abdominal aorta was 313.6 ± 29.6 in the A group and 322.4 ± 30.1 in the B group during the late arterial phase (p = 0.8). Meanwhile, the mean enhancement values of the portal vein were 176.2 ± 19.3 and 165.9 ± 24.5 in the A and B groups, respectively, during the portal venous phase (p = 0.6). The mean CT values of liver parenchyma were 117.1 ± 15.3 and 108.8 ± 18.7 in the A and B groups, respectively, during the portal venous phase (p = 0.9). There was no statistical difference in image quality, peak injection pressure (psi), and local discomfort between the two groups (p > 0.05). Conclusion: When a similar IDR and the same iodine load are used, CT images with different concentrations of contrast media have the same subjective and objective quality, and can meet the diagnostic needs.

Side-arm-promoted highly enantioselective ring-opening reactions and kinetic resolution of donor-acceptor cyclopropanes with amines.

A Ni-catalyzed asymmetric ring-opening reaction of 2-substituted cyclopropane-1,1-dicarboxylates with aliphatic amines has been accomplished using the chiral indane-trisoxazoline (In-TOX) ligand. This highly enantioselective reaction provides an efficient approach to a variety of chiral γ-substituted γ-amino acid derivatives, which are readily transformed into multifunctionalized piperidines and γ-lactams. The single-crystal X-ray structure of the TOX-Ni complex is provided, and the role of the side arm in the chiral ligand is discussed.

Malignant potential of oral and labial chronic discoid lupus erythematosus: a clinicopathological study of 87 cases.

AIMS: To investigate clinicopathological features and identify clinicopathological risk factors for the malignant transformation of oral and labial chronic discoid lupus erythematosus (DLE) in a relatively large number of patients from China. METHODS AND RESULTS: A total of 87 patients with clinical and histopathological diagnosis of DLE between 1993 and 2009 were reviewed retrospectively in our hospital. The average age at diagnosis was 51.7 years, with a male:female ratio of 1:1.8. The lower lip was the most common site (71.3%). We documented six DLE patients with malignant transformation. On univariate analysis, patients with high-risk dysplasia (P = 0.002) or aged >60 (P = 0.045) were associated with DLE malignant transformation, but gender, lesion site, smoking and alcohol intake were not risk factors. On multivariate analysis, high-risk dysplasia was a significant indicator for DLE malignant transformation. High-risk dysplasia was associated with a 14.24-fold [95% confidence interval (95% CI), 1.97-102.88; P = 0.008] increased risk of malignant transformation, compared with non/low-risk dysplasia. CONCLUSIONS: The utilization of high-risk dysplasia as a significant indicator for evaluating malignant transformation risk in patients with DLE is suggested, which may be helpful to guide treatment selection.

[Dynamics of three-dimensional ecological footprint of Zhejiang coastal zone and its influencing factors based on GTWR model]. / 基于GTWRæ¨¡åž‹çš„æµ™æ±Ÿçœæµ·å²¸å¸¦ä¸‰ç»´ç”Ÿæ€è¶³è¿¹åŠ¨æ€å˜åŒ–åŠå ¶å½±å“å› ç´ .

Clarifying the consumption of local natural capital stock and natural capital flow and its influencing factors can help alleviate the tension between the socio-economic development of the coastal zone, natural resources, and the ecological environment, and promote regional coordinated and sustainable development. Based on the two-dimensional and three-dimensional ecological footprint evaluation models, we analyzed the spatial and temporal variations of ecological footprint in the coastal zone of Zhejiang Province from 2006 to 2016, and explored the regional natural capital stock and capital flow occupancy based on footprint depth and footprint width. We analyzed the spatial and temporal heterogeneity of its influencing factors using a geographically and temporally weighted regression model. The results showed that ecological crisis in the coastal zone of Zhejiang Province deepened during the study period. The average per capita ecological deficit was 3.5 hm2·cap-1, and the per capita ecological deficit increased by 15.5%. Among them, the contribution of energy consumption account to the per capita ecological footprint was greater than that of the biological resources account. The per capita ecological footprint of each county showed an increasing trend, with significant spatial variation. The coastal zone of Zhejiang Province mainly consumed the natural capital stock, with a significant inhibitory effect on the renewal of natural capital flows. The consumption of natural capital stock was 14.87 times that of capital flow in 2016. Urban scale, economic development level, and people's income level had positive impacts on the ecological footprint of coastal counties of Zhejiang Province, while fishery production had negative effect on the ecological footprints, with transition trend to a positive one. The influence mechanism of the three factors had significant spatial and temporal differentiation.

Lipopolysaccharide induces IL-6 production in respiratory syncytial virus-infected airway epithelial cells through the toll-like receptor 4 signaling pathway.

Respiratory syncytial virus (RSV) is a leading cause of bronchiolitis in young children. Microbial agents such as endotoxin and RSV are implicated in airway inflammation during the development of reactive airway disease (RAD) later in childhood. Toll-like receptors (TLRs) are involved in an inflammation cascade through pathogen-associated molecular pattern recognition including lipopolysaccharide (LPS) and viral components. In this study, we investigated the expression of TLRs and cytokine-chemokine production profiles of RSV-infected epithelial cells. In live-RSV infected human tracheal epithelial cell line (9HTEo), TLRs 1-10 mRNA levels were up-regulated in a time-dependent manner compared with ultraviolet (UV)-inactivated RSV. RSV was shown to alter TLR4 membrane and cytosolic location in epithelial cells. Stimulating RSV-infected epithelial cells with TLR4 agonist LPS increased synthesis of IL-6, IL-8, and reduced regulated on activation, normal T cell expressed and secreted (RANTES) production. TLR4 neutralizing antibody HTA125 and TLR4-targeting RNA interference experiments revealed that TLR4 signaling pathway played a predominant role in mediating LPS-induced-IL-6 production of RSV infected epithelial cells. Altogether, our studies indicated that TLR4 play a critical role in leading LPS mediated-IL-6 response in RSV infected-epithelial cells and might be an important factor influencing the cytokine-chemokine profile of epithelial cells interacting with virus and endotoxin, which is correlated with phenotypes of RSV diseases.

Antidepressant effect of venlafaxine in chronic unpredictable stress: Evidence of the involvement of key enzymes responsible for monoamine neurotransmitter synthesis and metabolism.

A number of studies have linked abnormalities in the function of the serotonergic and noradrenergic systems to the pathophysiology of depression. It has been reported that selective serotonin reuptake inhibitors promote the expression of tryptophan hydroxylase (TPH), which is involved in the synthesis of serotonin. However, limited evidence of TPH alteration has been found in selective serotonin and noradrenaline reuptake inhibitors (SNRIs), and more key enzymes need to be investigated. The aim of the present study was to determine whether venlafaxine (VLX; a classical SNRI) regulates TPH and other key enzymes responsible for the synthesis and metabolism of monoaminergic transmitters in rats with chronic unpredictable stress (CUS). The present results suggested that CUS­exposed rats exhibited decreased locomotor activity in the open­field test and increased immobility time in the forced swim test, as compared with the controls. Pretreatment with VLX (20 mg/kg) significantly increased locomotor activity and reduced immobility time in the CUS­exposed rats. In addition, VLX (20 mg/kg) treatment prevented the CUS­induced reduction in tyrosine hydroxylase and TPH expression in the cortex and hippocampus. Furthermore, VLX alleviated the CUS­induced oxidative stress in the serum, cortex and hippocampus. However, VLX administration did not have an effect on indoleamine­2,3­dioxygenase overexpression in the hippocampus. It was therefore concluded that the regulation of abnormalities in the synthesis and metabolism of monoaminergic transmitters may be associated with the antidepressant effects of VLX, suggesting that multimodal pharmacological treatments can efficiently treat depression.

Antidepressant mechanisms of venlafaxine involving increasing histone acetylation and modulating tyrosine hydroxylase and tryptophan hydroxylase expression in hippocampus of depressive rats.

Venlafaxine (VEN) is a widely used antidepressant as a serotonin-reuptake and norepinephrine-reuptake inhibitor. It is used primarily in depression, especially with generalized anxiety disorder or chronic pain. This medicine is of interest because its mechanisms involved multiple aspects. In the current study, the antidepressant action of VEN was investigated by studying the histone acetylation and expression of tyrosine hydroxylase (TH) and tryptophan hydroxylase (TPH) in rats exposed to chronic unpredicted stress (CUS) for 28 days. Male Sprague-Dawley rats were divided into a control group, VEN-treated control group, CUS group, and VEN-treated CUS group. VEN (23.4 mg/kg once daily) was administered to rats by intragastric gavage, whereas the same volume of vehicle was given to rats in the control and model groups. Rat behaviors, acetylated H3 at lysine 9 (acH3K9), acetylated H3 at lysine 14 (acH3K14), acetylated H4 at lysine 12 (acH4K12), histone deacetylase 5, and TH and TPH expression in the hippocampus were determined. Chronic VEN treatment significantly relieved the anxiety- and depression-like behaviors, prevented the increase of histone deacetylase 5 expression and decrease of acH3K9 level, and promoted TH and TPH protein expression in the hippocampus of CUS rats. The results suggest that the preventive antidepressant mechanism of VEN is partly involved in the blocking effects on histone de-acetylated modification and then increasing TH, TPH expression.

[Toll-like receptor 4 expression and function of respiratory syncytial virus-infected airway epithelial cells].

OBJECTIVE: To observe the epithelial Toll like receptor (TLR)4 expression changes and the signaling pathway function after respiratory syncytial virus (RSV) infection and to explore the mechanisms of RSV-induced airway inflammation. METHODS: 9HTEo-human tracheal epithelial cell line was infected by RSV (MOI = 10), and TLR1-10 mRNA were detected by RT-PCR assay at 3 h post RSV infection. TLR4 mRNA was detected by real time Q-PCR assay at 3 h, 6 h and 9 h post RSV infection, and TLR4 protein expression and cell apoptosis were determined by flow cytometry at 24 h post RSV infection. IL-8 in supernatant was detected by ELISA after RSV-infected cells exposed to lipopolysaccharide (LPS). A normal control group and a RSV infection group were set up for the RT-PCR and flow cytometry experiments, and the data were analyzed by paired t test using GraphPad 4.0 software. A normal group, a RSV group and a UV-inactivated RSV group were set up for the real time Q-PCR, experiments, and the data were analyzed by Kruskal-Wallis test. The ELISA experiments were divided into 4 groups including a normal control, a RSV, a LPS stimulation, and a RSV plus LPS co-stimulation groups, and the data were analyzed by One-way ANOVA test. RESULTS: (1) TLR2-10 mRNA level was significantly up-regulated (t value of TLR2-10: 3.49 -14.47, P < 0.05), especially TLR-2, 6 enhanced expression, compared with the normal epithelial cells. Real time Q-PCR assay showed that TLR4 mRNA started to increase at 3hr (Kruskal-Wallis test value = 8.82, P < 0.05, n = 6) and significantly elevated at 9 hour (Kruskal-Wallis test value = 6.62, P < 0.05, n = 6). UV inactivated-RSV had no effect on the TLR4 mRNA level. (2) Flow cytometry showed that membrane TLR4 mean fluorescence intensity (MFI) increased (RSV: 1.27 +/- 0.48, normal: 0.97 +/- 0.25; t = 2.39, P > 0.05, n = 10) while cytoplasmic TLR4 MFI simultaneously decreased (RSV: 3.08 +/- 1.38, normal: 3.36 +/- 1.31, t = 2.92, P = 0.225, n = 10). Percentage of membrane TLR4-positive cells was higher in RSV infected population [RSV: (11.99 +/- 7.74)%, normal: (1.16 +/- 0.47)%, Mann-Whitney t value = 0.001, P < 0.01, n = 8], most (93.32 +/- 1.7)% of which were Annexin V positive. IL-8 was significantly induced in the RSV plus LPS costimulation group compared with RSV group (F = 59.29, P < 0.01, n = 3). CONCLUSIONS: RSV induced epithelial TLR4 up-regulation, localization changes from cytoplasm to membrane, IL-8 secretion through TLR4 signaling pathway and epithelial cell apoptosis in membrane TLR4 positive population. These results indicate TLR4 is involved in RSV-induced acute or chronic epithelial-dependent inflammation, which might contribute to acute or chronic airway inflammation.

Automatic detection of pulmonary nodules in CT images by incorporating 3D tensor filtering with local image feature analysis.

Computer-aided detection (CAD) technology has been developed and demonstrated its potential to assist radiologists in detecting pulmonary nodules especially at an early stage. In this paper, we present a novel scheme for automatic detection of pulmonary nodules in CT images based on a 3D tensor filtering algorithm and local image feature analysis. We first apply a series of preprocessing steps to segment the lung volume and generate the isotropic volumetric CT data. Next, a unique 3D tensor filtering approach and local image feature analysis are used to detect nodule candidates. A 3D level set segmentation method is used to correct and refine the boundaries of nodule candidates subsequently. Then, we extract the features of the detected candidates and select the optimal features by using a CFS (Correlation Feature Selection) subset evaluator attribute selection method. Finally, a random forest classifier is trained to classify the detected candidates. The performance of this CAD scheme is validated using two datasets namely, the LUNA16 (Lung Nodule Analysis 2016) database and the ANODE09 (Automatic Nodule Detection 2009) database. By applying a 10-fold cross-validation method, the CAD scheme yielded a sensitivity of 79.3% at an average of 4 false positive detections per scan (FP/Scan) for the former dataset, and a sensitivity of 84.62% and 2.8 FP/Scan for the latter dataset, respectively. Our detection results show that the use of 3D tensor filtering algorithm combined with local image feature analysis constitutes an effective approach to detect pulmonary nodules.

Inhibition of respiratory syncytial virus of subgroups A and B using deoxyribozyme DZ1133 in mice.

Respiratory syncytial virus (RSV) commonly infects the upper and lower respiratory tracts. Currently, there is no effective treatment available. Deoxyribozymes are a potential therapeutic for RSV and their activity is based on the ability to bind and cleave complementary RNA sequences to inhibit protein expression. DZ1133 is a deoxyribozyme that targets the conserved genomic RNA sequence of the RSV nucleocapsid protein and has been shown to significantly inhibit various strains of RSV including subgroups A and B, standard A2 and CH18537 strains, and CQ381513, CQ381170, BJ01 and BJ04 strains. Treatment with DZ1133 decreased viral plaque formation in lungs of RSV-infected BALB/c mice. In addition, viral mRNA expression was reduced, airway inflammation was alleviated, and leukocyte counts were reduced in bronchoalveolar lavage fluid of RSV-infected mice. The antiviral effect of DZ1133 was dose-dependent (0.2-0.8mg) and more efficient than antisense oligonucleotide inhibition of gene expression. However, levels of cytokines TNF-alpha, IFN-gamma, IL-12, and IL-10 induced by RSV infection were not affected by DZ1133 treatment. Our data demonstrate that DZ1133 is a potential therapeutic agent against both subgroups A and B RSV infection in vivo.

Inhibition of respiratory syncytial virus in cultured cells by nucleocapsid gene targeted deoxyribozyme (DNAzyme).

Respiratory syncytial virus (RSV), which presents the primary cause of bronchiolitis and pneumonia among infants and causes significant morbidity and mortality in immunodeficient patients, remains a health problem worldwide. Unfortunately, an effective vaccine is currently unavailable and pharmacologic treatment needs further optimization for RSV disease. Because RSV is a non-segmented negative-strand RNA virus, it may be sensitive to the genome RNA cleaving by DNAzyme, an artificial nucleic acids molecule with high catalytic capability of cleaving complementary RNA molecules. Thus, RSV-targeted DNAzymes potentially present as a therapeutic candidate of RSV diseases. In this study, DNAzymes targeting the RSV genomic RNA or mRNA were designed and synthesized, one of which (DZn1133) did cleave RSV RNA in vitro, inhibit the transcription and expression of F viral gene, reduce the RSV yield by about 7 logs and protect more than 90% RSV-infected Hep-2 cells from a cytopathic effect at 8 microM. Moreover, 10 wild RSV strains isolated from clinic patients including both subgroups A and B were all suppressed by DZn1133 with greater anti-RSV activity than antisense DNA or ribavirin.