Evaluation of enterovirus nucleic acids detection method based on ultra-fast real-time fluorescence RT-PCR technology-A pilot study.

The outbreak of COVID-19 epidemic has enabled the establishment and application of various rapid detection methods. It is particularly important to establish a fast and accurate detection method for enterovirus, which will be beneficial for clinical diagnosis, epidemic prevention and control, and timely traceability. Through establishing an ultra-fast reverse transcription-polymerase chain reaction (RT-PCR) equipment, this study aimed to evaluate the sensitivity and specificity of the testing method of enterovirus nucleic acids based on ultra-fast real-time fluorescence RT-PCR technology. A total of 61 cases were sampled, which were then transported and preserved. After the nucleic acid extraction, the nucleic acids of the same sample were tested with the enterovirus nucleic acid detection kit produced by Guangzhou Da An Gene Company and the ultra-fast RT-PCR equipment system established in this study. ABI7500Fast and Ahram biosystems S1 fast equipment were used for amplification detection. If the sample had an S-shaped amplification curve in the FAM channel and the Ct value ≤40.00, the result was positive. The sensitivity, precision, and accuracy of the detection method were then verified. This study established a novel testing method to achieve enterovirus nucleic acid detection within 24 min. The sensitivity detection limit of the method was 1.0 × 102 copies/ml. The coefficients of variation for repeated detection of the high, medium, and low concentration samples were 2.644%, 1.674%, and 4.281%, respectively, with good detection repeatability. In addition, a total of 29 cases were positive by the ultra-fast RT-PCR detection method in 61 suspected samples, which was consistent with the conventional fluorescent RT-PCR method. The established rapid detection method can greatly shorten the time for providing a detection report, which may greatly improve the efficiency of diagnosis and treatment.

Integrated Network Pharmacology and Experimental Validation Approach to Investigate the Therapeutic Effects of Capsaicin on Lipopolysaccharide-Induced Acute Lung Injury.

An integrated method combining network pharmacology and in vivo experiment was performed to investigate the therapeutic mechanism of capsaicin (Cap) against acute lung injury. The potential key genes and signaling pathways involved in the therapeutic effect of Cap were predicted by the network pharmacology analyses. Additionally, the histological assessment, ELISA, and RT-qPCR were performed to confirm the therapeutic effect and the potential mechanism action involved. Our findings showed that TNF, IL-6, CXCL1, CXCL2, and CXCL10 were part of the top 50 genes. Enrichment analysis revealed that those potential genes were enriched in the TNF signaling pathway and IL-17 signaling pathway. In vivo experiment results showed that Cap alleviated histopathological changes, decreased inflammatory infiltrated cells and inflammatory cytokines, and improved antioxidative enzyme activities in the bronchoalveolar lavage fluid (BALF). Furthermore, Cap treatment effectively downregulated TNF, IL-6, NF-κB, CXCL1, CXCL2, and CXCL10 in lung tissue. Thus, our findings demonstrated that Cap has the therapeutic effect on LPS-induced acute lung injury in neonatal rats via suppression of the TNF signaling pathway and IL-17 signaling pathway.

Changes in humoral immunity, myocardial damage, trace elements, and inflammatory factor levels in children with rotavirus enteritis.

OBJECTIVE: To investigate the changes and significance of humoral immunity, myocardial damage, trace elements and inflammatory factors levels in children with rotavirus enteritis. METHODS: One hundred children with rotavirus enteritis admitted to our hospital from January 2019 to December 2020 were retrospectively selected as the case group, and they were divided into a no dehydration group (33 cases), mild dehydration group (41 cases), and moderate dehydration group (26 cases). Another 100 children with rotavirus-negative enteritis during the same period were selected as the control group. Serum immunoglobulin, cardiac enzyme profile, trace elements, and interleukin-6 (IL-6) levels were compared between the two groups, and among the case groups for different degrees of dehydration. RESULTS: Serum immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM), zinc, magnesium, and calcium in the case group were lower than in controls (P<0.05). Serum lactate dehydrogenase (LDH), α-hydroxybutyrate dehydrogenase (α-HBDH), creatine kinase (CK), and creatine kinase isoenzyme (CK-MB) in the case group were higher than in controls (P<0.05). Serum IL-6, interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) were also higher in cases than controls (P<0.05). Serum IgA, IgG, IgM, zinc, magnesium, and calcium in children with rotavirus enteritis with mild dehydration were lower than those without dehydration, but higher than those with moderate dehydration (P<0.05). Serum LDH, α-HBDH, CK, and CK-MB in children with rotavirus enteritis with mild dehydration were higher than those without dehydration, but lower than those with moderate dehydration (P<0.05). Serum IL-6, IL-8, and TNF-α in children with rotavirus enteritis with mild dehydration were higher than those without dehydration, but lower than those with moderate dehydration (P<0.05). CONCLUSION: Children with rotavirus enteritis with more severe dehydration exhibited lower levels of humoral immunity and trace elements and greater myocardial damage and inflammatory response. Early detection can accurately assess the condition and provide a reference for clinical treatment.

Congenital cytomegalovirus infection and the risk of hearing loss in childhood: A PRISMA-compliant meta-analysis.

BACKGROUND: Congenital cytomegalovirus (cCMV) infection is the most common cause of childhood hearing loss (HL), although the strength of this association remains limited and inconclusive. Thus, the purpose of this study was to summarize evidence regarding the strength of the relationship between cCMV and childhood HL and to determine whether this relationship differs according to patient characteristics. METHODS: The PubMed, EmBase, and Cochrane Library databases were searched for studies evaluating the relationship between cCMV and HL from inception to September 2019. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were used to calculate the investigated outcomes in a random-effects model. Sensitivity, subgroup, and publication bias analyses were also performed. RESULTS: A total of 15 studies involving 235,026 children met the inclusion criteria and were included in the final analysis. The summary results indicated that cCMV infection was associated with an increased risk of HL (odds ratio [OR]: 8.45; 95% confidence interval [CI]: 3.95-18.10; P < .001), irrespective of whether studies reported sensorineural HL (OR: 5.42; 95% CI: 1.98-14.88; P = .001), or did not evaluate HL types among their patients (OR: 11.04; 95% CI: 3.91-31.16; P < .001). However, in studies conducted in the United States (P < 0.001) and published in or after 2000 (P = 0.026), the study populations included <60% males (P < 0.001). Moreover, studies of high quality (P < .001) demonstrated a significantly greater risk of HL with cCMV infection than that in the corresponding subgroups. CONCLUSIONS: The study results suggest that cCMV infection increases the risk of HL. Further studies are required to investigate the association of cCMV infection with the risk of specific subtypes of HL.

Alteration in autophagy gene expression profile correlates with low sperm quality.

AIMS: Low sperm quality, a crucial factor of male infertility, is becoming a public health issue all over the world. In male reproductive system, autophagy plays an important role in maintaining physiological functions. There exist conjectures that disordered autophagy autophagy might be related to low sperm quality. However, there is no evidence can confirm that. This study aims to investigate the association between autophagy-associated genes and low sperm quality. METHODS: Sperm samples of low sperm quality cases and matched controls were included to select differential expressed genes (DE genes) by autophagy-related functional gene microarray analysis. Then, 104 cases and 250 controls were included to validate the expression of four important autophagy genes (CXCR4, ESR1, PTEN and LC3B). Based on the obtained DE gene, gene Ontology and pathway analyses were conducted. RESULTS: Chip results showed that expression of all 18 DE genes were decreased in the cases compared to the controls (P < 0.05). Expression of ESR1 were verified to be significantly decreased (P < 0.05). CONCLUSION: Our results provided clues with the association among down-regulated expression of autophagy regulating and associated genes and low sperm quality. These findings revealed possible role of impaired autophagy in the mechanism of low sperm quality. Moreover, these may also provide potential targets for the treatment of low sperm quality.

SCN1A IVS5N+5 G>A Polymorphism and Risk of Febrile Seizure and Epilepsy: A Systematic Review and Meta-Analysis.

Background: Previous studies had investigated the association between polymorphism of IVS5N+5 G>A in SCN1A and the risk of febrile seizure and epilepsy. However, the results were inconsistent. We aimed to conduct a systematic review and meta-analysis to evaluate the association between SCN1A IVS5N+5 G>A polymorphism and risk of febrile seizures and epilepsy. Methods: We searched Embase, Medline, Scopus, and CNKI for studies on the association between SCN1A IVS5N+5 G>A polymorphism and risk of febrile seizures and epilepsy up to 19 February 2020. We pooled odds ratios (ORs) and 95% confidence intervals (CIs) by different genetic models. To explore the source of heterogeneity, we performed the subgroup analysis by ethnicity and source of control. Results: We included a total of 12 studies in the meta-analysis. We found a significant negative association between G allele SCN1A IVS5N+5 G>A polymorphism, febrile seizures [G vs. A: OR (95% CI): 0.690 (0.530-0.897); GG vs. AA: 0.503 (0.279-0.908); AG vs. AA: 0.581 (0.460-0.733); GG + AG vs. AA: 0.543 (0.436-0.677); AA + GG vs. AG: 1.309 (1.061-1.615)], and epilepsy [G vs. A: 0.822 (0.750-0.902); GG vs. AA: 0.655 (0.515-0.832); AG vs. AA: 0.780 (0.705-0.862); GG vs. AG + AA: 0.769 (0.625-0.947); GG + AG vs. AA: 0.743 (0.663-0.833); AA + GG vs. AG: 1.093 (1.001-1.193)]. The subgroup analysis shows the association varied by type of disease, ethnicity, and source of control. Conclusion: The present meta-analysis suggests that G allele in SCN1A IVS5N+5 G>A polymorphism is a protective factor of febrile seizures and epilepsy. It is possible to determine the vulnerability of each individual to develop febrile seizures or epilepsy genotype by these genetic variants. Future studies with better study designs are needed to confirm the results. Study Registration: This study was registered in the International Prospective register of systematic reviews (PROSPERO, CRD42020163318).

Eupatilin Alleviates Lipopolysaccharide-Induced Acute Lung Injury by Inhibiting Inflammation and Oxidative Stress.

BACKGROUND Eupatilin, an active flavone separated from Artemisia species, has various biological activity such as anti-inflammatory activity. The aim of the present study was to find out the influence of eupatilin on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. MATERIAL AND METHODS The administration of LPS was used to induce ALI; eupatilin was given 1 hour before the LPS administration. Lung structural damage of rats was analyzed by hematoxylin and eosin staining and the wet/dry lung ratio. The related inflammatory factors and lung injury markers were examined by enzyme-linked immunosorbent assay. The oxidative stress factors were analyzed by corresponding kits. The expression of peroxisome proliferator-activated receptor-alpha (PPAR-alpha) was assayed by western blot and immunohistochemical staining. RESULTS The results showed that eupatilin alleviated LPS-induced structural damage and decreased the wet/dry lung ratio concentration-dependently. Eupatilin decreased the level of surfactant protein (SP)-A, SP-D, and inflammatory factors such as interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, and monocyte chemo-attractant protein (MCP)-1. LPS trigged nitric oxide (NO) generation, improved the production of malondialdehyde (MDA) and lactate dehydrogenase (LDH) and decreased the activity of superoxide dismutase (SOD), which were reversed when rats treated with eupatilin in a concentration-dependent way. Besides, the expression of PPAR-a was increased under the treatment of eupatilin. CONCLUSIONS Collectively, eupatilin alleviated LPS-induced ALI through inhibiting inflammation and oxidative stress in a concentration-dependent way, which was likely to be closely related with the activation of PPAR-alpha.