BackgroundThe aim of this study is to examine the safety and distribution of Ad-EGFP-MDR1, an adenovirus encoding human multidurg resistance gene (human MDR1), in the mice colon carcinoma model.MethodsAfter bone marrow cells (BMCs) were infected with Ad-EGFP-MDR1, they were administered by intra bone marrow-bone marrow transplantation (IBM-BMT). Total adenovirus antibody and serum adenovirus neutralizing factor (SNF) were determined. Biodistribution of Ad-EGFP-MDR1 was detected by in situ hybridization and immunohistochemistry. The peripheral hematocyte white blood cell (WBC), haemoglobin (Hb), red blood cell (RBC) and platelet (Plt) counts were analyzed.ResultsNeither total adenovirus antibody nor SNF increased weeks after BMT. In situ hybridization and immunohistochemistry demonstrated concordant expression of human MDR1 and P-gp which were found in lung, intestine, kidney and BMCs after BMT, but not detected in liver, spleen, brain and tumor. No significant abnormality of the recovery hematocyte was observed on Day 30 after treatment.ConclusionThe results indicate that IBM-BMT administration of a replication defective adenovirus is a feasible mode of delivery, allowing exogenous transference. The findings in this study are conducted for the future long-term studies of safety assessment of Ad-EGFP-MDR1.

Hand-foot-and-mouth disease (HFMD) is a common childhood infection that may lead to serious complications and even death. Globally, epidemics of HFMD are increasing each year, especially in China. This study aimed to identify risk factors for death in children with critical and severe HFMD in Chongqing, China.We performed an observational study involving patients with critical and severe HFMD admitted to the Children's Hospital of Chongqing Medical University from January 2009 to December 2016. Overall, 179 patients aged 2 months to 16 years, were included; 127 died (non-survival group) and 52 survived (survival group); the case-fatality rate was 70.94%. Data comprising demographic characteristics, clinical symptoms and signs, and laboratory findings were collected. Non-conditional logistic regression analysis was performed to determine the risk factors for death.Univariate analysis showed that sex, coma, light-reflex insensitivity, pulmonary rales, pulmonary edema or hemorrhage, cold extremities, tachycardia, hypotension, white blood cell count, blood glucose concentration, serum lactate level, creatine kinase-MB isoenzyme level, and acidosis were associated with death (P < .05). Logistic regression analysis identified female sex (odds ratio [OR] 9.6, 95% confidence interval [CI] 3.0–30.2), light-reflex insensitivity (OR 4.4, 95% CI 1.4–13.1), tachycardia (OR 1.05, 95% CI 1.03–1.07), and higher serum lactate levels (OR 1.14, 95% CI 1.19–1.69) as independent risk factors; and longer onset-to-hospitalization time (OR 0.43, 95% CI 0.28–0.66) as an independent protective factor for death in children with critical and severe HFMD.Female sex, light-reflex insensitivity, tachycardia, and higher serum lactate level are potential independent risk factors; and longer onset-to-hospitalization time is possibly an independent protective factor for death in patients with critical and severe HFMD.

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ObjectiveA novel multi-drug resistance gene named as HA117 has been screened and cloned in multidrug resisitant leukemia cell lines in our previous research, but its function is still unknown. In this study, HA117 gene was investigated whether it could increase the drug resistance in chronic myelogenous myeloid leukemia cell line K562.MethodsHA117 was cloned and adenovirus vectors were constructed with the HA117 gene (Adeasy-HA117). K562 cells were infected by Ad-HA117 to get the K562/Ad-HA117 cells with HA117 gene expression. The infection efficiency and the multiplicity of infection (MOI) were detected by fluorescence and flow cytometry. The expression of HA117 gene was detected by RT-PCR. The drug sensitivities of K562/Ad-HA117 cells were detected by Methyl Thiazolyl Tetrazolium (MTT) assay.ResultsRecombinant adenovirus vectors were constructed and a MOI of 100 is most suitable to infect K562 cells. The infected K562 cells demonstrated in vitro production of HA117 mRNA as measured by reverse-transcriptase polymerase chain reaction. There were no significant changes in K562/Ad-HA117 cells growth, while the drug sensitivities of K562/Ad-HA117 cells to Vincristine, Adriamycin, Etoposide, Daunorubicin, Mitomycin and Cyclophosphamide decreased 4.44, 7.18, 3.01, 9.53, 3.48 and 3.61 times than that of uninfected K562 cells, respectively (P < 0.05).ConclusionExpression of the novel gene HA117 could significantly increased the multi-drug resistance of K562 cells, which indicated that HA117 is a functionally relevant multidrug resistance gene.

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A novel gene, HA117, was discovered in our previous work. Using the pSOS-HUS vector method which we designed at previous study, we screened for small interfering RNAs (siRNAs) that targeted HA117. The pSOS-HUS siRNA screening results were verified and a delivery system was developed that contained a recombinant adenovirus carrying DNA templates for the transcription of the HA117 siRNAs. Of five pairs of DNA templates, siRNA transcribed from HAi5 produced the strongest effect against HA117. A recombinant adenovirus containing HAi5 (Ad-HAi5) was successfully constructed and evaluated. This work has laid the foundation for further study of HA117 gene function using RNA interference technology and has showed the pSOS-HUS vector method was successfully utilized as a rapid and effective screen of siRNAs for a target gene.

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Aim: In this study, the protective effects of atorvastatin calcium (AC) on nerve cells and cognitive improvement in vivo and in vitro were investigated by establishing cell models and vascular dementia (VD) rat models. Background: VD is a neurodegenerative disease characterized by cognitive deficits caused by chronic cerebral hypoperfusion. AC has been studied for its potential to cure VD but its efficacy and underlying mechanism are still unclear. Objective: The mechanism of action of AC on cognitive deficits in the early stages of VD is unclear. Here, the 2-vessel occlusion (2-VO) model in vivo and the hypoxia/reoxygenation (H/R) cell model in vitro was established to investigate the function of AC in VD. Methods: The spatial learning and memory abilities of rats were detected by the Morris method. The IL-6, tumour necrosis factor-α (TNF-α), malondialdehyde (MDA) and superoxide dismutase (SOD) in cell supernatant was tested by ELISA kits. After behavioural experiments, rats were anaesthetized and sacrificed, and their brains were extracted. One part was immediately fixed in 4% paraformaldehyde for H&E, Nissl, and immunohistochemical analyses, and the other was stored in liquid nitrogen. All data were shown as mean ± SD. Statistical comparison between the two groups was performed by Student’s t-test. A two-way ANOVA test using GraphPad Prism 7 was applied for escape latency analysis and the swimming speed test. The difference was considered statistically significant at p < 0.05. Results: AC decreased apoptosis, increased autophagy, and alleviated oxidative stress in primary hippocampal neurons. AC regulated autophagy-related proteins in vitro by western blotting. VD mice improved cognitively in the Morris water maze. Spatial probing tests showed that VD animals administered AC had considerably longer swimming times to the platform than VD rats. H&E and Nissl staining showed that AC reduces neuronal damage in VD rats. Western blot and qRT-PCR indicated that AC in VD rats inhibited Bax and promoted LC3-II, Beclin-1, and Bcl-2 in the hippocampus region. AC also improves cognition via the AMPK/mTOR pathway. Conclusion: This study found that AC may relieve learning and memory deficits as well as neuronal damage in VD rats by changing the expression of apoptosis/autophagy-related genes and activating the AMPK/mTOR signalling pathway in neurons.