Search

Viral encephalitis is an infectious disease severely affecting human health. It is caused by a wide variety of viral pathogens, including herpes viruses, flaviviruses, enteroviruses, and other viruses. The laboratory diagnosis of viral encephalitis is a worldwide challenge. Recently, high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections. Thus, In this study, we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. We designed nine pairs of specific polymerase chain reaction (PCR) primers for the 12 viruses by reviewing the relevant literature. The detection ability of the primers was verified by software simulation and the detection of known positive samples. Amplicon sequencing was used to validate the samples, and consistency was compared with Sanger sequencing. The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×, and the sequence lengths were consistent with the sizes of the predicted amplicons. The sequences were verified using the National Center for Biotechnology Information BLAST, and all results were consistent with the results of Sanger sequencing. Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis. It is also a useful tool for the high-volume screening of clinical samples. [ABSTRACT FROM AUTHOR]

Japanese Encephalitis in China in the Period of 1950-2018: From Discovery to Control. [Extracted from the article]

Objective: Genotypes (G) 1, 3, and 5 of the Japanese encephalitis virus (JEV) have been isolated in China, but the dominant genotype circulating in Chinese coastal areas remains unknown. We searched for G5 JEV-infected cases and attempted to elucidate which JEV genotype was most closely related to human Japanese encephalitis (JE) in the coastal provinces of China., Methods: In this study, we collected serum specimens from patients with JE in three coastal provinces of China (Guangdong, Zhejiang, and Shandong) from 2018 to 2020 and conducted JEV cross-neutralization tests against G1, G3, and G5., Results: Acute serum specimens from clinically reported JE cases were obtained for laboratory confirmation from hospitals in Shandong (92 patients), Zhejiang (192 patients), and Guangdong (77 patients), China, from 2018 to 2020. Seventy of the 361 serum specimens were laboratory-confirmed to be infected with JEV. Two cases were confirmed to be infected with G1 JEV, 32 with G3 JEV, and two with G5 JEV., Conclusion: G3 was the primary infection genotype among JE cases with a definite infection genotype, and the infection caused by G5 JEV was confirmed serologically in China., (Copyright © 2024 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

KN motif and ankyrin repeat domains 1 (Kank1) and ki67 are associated with tumorigenesis and progression. This paper researched the expression of Kank1 and Ki67 and their clinicopathologic significance in pulmonary adenocarcinoma (PA). We monitored the expression of KanK1 and ki67 in 94 cases of human PA and 31 cases of paracancerous tissue by the immunohistochemical method. The results showed that Kank1 protein was detected in 74.2% (41/94) of PA tissues, and they were associated with differentiation (P = 0.025) and lymphatic metastasis (P = 0.002). Kaplan-Meier analysis suggested that patients with low Kank1 expression had shorter overall survival in PA (P = 0.020). Ki67 protein was detected in 79.8% (75/94) of PA tissues, and they were associated with differentiation (P < 0.001), TNM classification (P = 0.007), and lymphatic metastasis (P = 0.044). Furthermore, Kaplan-Meier analysis showed that patients with overexpression of Ki67 had shorter overall survival (P = 0.014). Cox multivariate analysis showed that tumor differentiation, TNM classification, lymphatic metastasis, Kank1, and ki67 expression were independent factors for prognosis of PA (P = 0.012, 0.016, 0.007, 0.021 and P = 0.003 respectively). In conclusion, compared with paracancerous tissues, Kank1 had low expression, while Ki67 was overexpressed in PA. They are closely related to its occurrence and development, and the prognosis of patients with low expression of Kank1 or overexpression of ki67 was poor in PA. Kank1 and Ki67 can be helpful for diagnosing and detecting the prognosis of patients with PA., Competing Interests: None., (IJCEP Copyright © 2020.)

Progesterone and adipoQ receptor family member 3 (PAQR3) and vascular endothelial growth factor (VEGF)-A are associated with tumorigenesis and progression. The aim of this study is to investigate the expression of PAQR3 and VEGF-A in pulmonary adenocarcinoma (PA) and explore their clinical and pathologic significance. The expressions of PAQR3 and VEGF-A protein were detected in 86 cases of human PA and 26 cases of tumor-adjacent tissue by immunohistochemistry. The positive rate of PAQR3 was 39.5% in PA, which was lower than that in tumor-adjacent tissues (80.8%), P=0.001. Negative expression of PAQR3 was obviously linked to tumor TNM stage, differentiation, and lymphatic metastasis; and P values were 0.013, 0.025, and 0.034, respectively. The positive expression rate of VEGF-A was 68.6% in human PA whichwas higher than that of tumor-adjacent tissues (11.5%), P<0.001. The positive expression of VEGF-A was correlated with tumor TNM stage, differentiation, and lymphatic metastasis, and P values were 0.026, 0.001 and P=0.001, respectively. The expression of PAQR3 was negatively correlated with the expression of VEGF-A (r=-0.698, P<0.001). Log-rank test statistical analysis suggested that patients with negative expression of PAQR3 or positive expression of VEGF-A had shorter overall survival. Cox multivariate analysis indicated that tumor TNM stage, differentiation, and lymphatic metastasis, and PAQR3 and VEGF-A expression were independent factors for prognosis of PA, and P values were 0.021, 0.017, 0.006, 0.018 and P=0.007 respectively. In conclusion, negative expression of PAQR3 and positive expression of VEGF-A are markedly correlated with tumor TNM classification, histologic grade, and lymphatic metastasis. Tumor TNM stage, differentiation, and lymphatic metastasis, negative expression of PAQR3, and positive expression of VEGF-A are risk factors for prognosis of patients with PA. Detection of PAQR3 and VEGF-A may be helpful to evaluate prognosis and infiltrative capability of PA., Competing Interests: None., (IJCEP Copyright © 2020.)

Astrocyte elevated gene-1 (AEG-1) and E-cadherin are associated with tumorigenesis and progression. The aim of this study is to investigate the expression of AEG-1 and E-cadherin in human gallbladder cancer (GBC) and explore their clinical and pathological significance. The expression of AEG-1 and E-cadherin protein were detected in 71 cases of human GBC and 22 cases of tumor-adjacent tissue by the immunohistochemical method. Our results demonstrate that the positive expression (high expression) rate of AEG-1 was 62.0% in human GBC which was higher than that in tumor-adjacent tissues (13.6%), P<0.001. The positive expression of AEG-1 protein was correlated with tumor TNM classification, histologic grade, and lymph node metastasis (P=0.037, P=0.033 and P=0.020, respectively). The positive expression rate of E-cadherin was 40.8% in GBC, which was lower than that in tumor-adjacent tissues (77.3%), P=0.003. Negative expression (Low expression) of E-Cadherin was significantly related with tumor TNM classification, histologic grade and lymphatic metastasis (P=0.028, P=0.003 and P=0.040, respectively). The expression of AEG-1 was negatively correlated with the expression of E-Cadherin (r=0.530, P<0.001). The log-rank test statistical analysis suggested that patients with positive expression of AEG-1 or negative expression of E-Cadherin protein had shorter overall survival time. Cox multivariate analysis showed that tumor TNM classification, histologic grade and lymphatic metastasis, AEG-1 and E-cadherin expression were independent factors for prognosis of GBC (P=0.013, P=0.019, P=0.001, P=0.011 and P=0.025 respectively). In conclusion, positive expression of AEG-1 and negative expression of E-Cadherin are markedly correlated with tumor TNM classification, histologic grade and lymphatic metastasis. The expression of AEG-1 was negatively correlated with the expression of E-Cadherin. Cox multivariate analysis showed that tumor TNM classification, histologic grade and lymphatic metastasis, positive expression of AEG-1 and negative expression of E-Cadherin were risk factors for prognosis of GBC. Detection of AEG-1 and E-Cadherin may be helpful to evaluate prognosis and infiltrative capability of gallbladder carcinoma., Competing Interests: None., (IJCEP Copyright © 2018.)

Objectives Pulmonary sequestration is a rare developmental abnormality of the lower respiratory system. This study aimed to evaluate the effectiveness of wedge resection compared with lobectomy for the treatment of intralobar pulmonary sequestration. Methods Video-assisted thoracic surgery (VATS) for intralobar pulmonary sequestration was performed in 26 patients in our institute between December 2006 and January 2015. Data regarding patient demographics, major complaints, diagnostic procedures, operative treatment, and treatment outcome were retrospectively analyzed. Results VATS was performed successfully in all patients. Wedge resection was performed in 7 patients and lobectomy in 19 patients. Conversion to thoracotomy was not required in any case. Statistical analysis revealed that operation duration and blood loss with wedge resection were significantly less than with lobectomy (p = 0.032 and 0.014, respectively). No significant differences were found in the mean drainage time, postoperative length of hospital stay, or complications. During our long-term follow-up, no patients had chronic cough, bloody sputum, or pneumonia. Conclusion VATS for intralobar pulmonary sequestration is feasible and safe. Lobectomy is the generally accepted operative method. However, wedge resection is a feasible alternative to lobectomy in select cases., (Georg Thieme Verlag KG Stuttgart · New York.)

The purpose of this article is to research on whether MACC1 can serve as a potential target for gene therapy of human bladder urothelial carcinoma (BUC). In this study, the expression of MACC1 gene was knocked down by RNA interference (RNAi) in the T24 cell (human BUC cell). The transcription level of MACC1 was detected by RT-PCR. Activities of MACC1, caspase-3, caspase-8, Bax and Met (mesenchymal-epithelial transition factor) protein were measured by Western blot. The cell proliferation and apoptosis were detected by MTT and flow cytometry. The cell's invasion ability was performed on Matrigel transwell assay. We also detect MMP2 (metalloproteinase-2) proteins by ELISA. The results showed that the level of MACC1 mRNA and protein was significantly reduced after RNAi. MTT assay showed that the proliferation of T24 cell was decreased due to RNA interference. Apoptosis studies also showed that MACC1 gene interference in T24 loses its anti-apoptotic effects. The expression of apoptosis proteins (Caspase-3, Caspase-8 and Bax) increased significantly due to the MACC1 RNAi. The level of Met protein was down-regulated obviously due to RNAi. Transwell assay showed that invasion abilities of T24 cells were reduced obviously due to MACC1 RNAi. Further studies showed that the secretion of MMP-2 was reduced by RNAi. It can conclude that the ability of proliferation and invasion in T24 cells can be inhibited by RNAi-targeting MACC1. As a result, MACC1 can serve as a potential target for gene therapy of human bladder urothelial carcinoma.

Osteopontin (OPN) involves in tumor formation, and strongly correlated with the tumor progression. It was overexpressed in human esophageal squamous cell carcinoma (ESCC). To study the molecular mechanisms of OPN in ESCC, we examined its roles in inhibiting proliferation and invasion of ECA-109 (esophageal squamous cell carcinoma) cells. The expression of OPN gene was knockdown by RNA interference (RNAi) in the Eca-109 cell. The transcription level of OPN was to detect by reverse transcription-quantitative PCR (RT-qPCR). Western blot assay was performed to detect the expression of OPN, Caspase-3,Caspase-8, Caspase-9, ERK1/2, phospho-ERK1/2 and MMP2 after RNAi. The cell proliferation and apoptosis were detected by MTT and Hoechst33342 assay. Transwell inserts was used for detecting ECA-109 cell's migration ability. The results shown that the level of OPN mRNA and protein was significantly reduced after RNAi. Proliferation and migration of cell line (ECA-109) was significantly inhibited in vitro. The protein phosphorylation and activation of ERK1/2 in the OPN RNAi group reduced significantly than the negative control groups. In Conclusion, the proliferation and migration of human ESCC can be inhibited by RNAi-targeting OPN. OPN can promote the expression of MMP2 through the ERK signaling pathways. OPN could serve as a potential therapeutic target for human ESCC.

To investigate the genetic characterization of Human parainfluenza virus-3 (HPIV-3) circulating in Gansu and Shaanxi Provinces of China, 719 throat swabs were collected from pediatric patients with acute respiratory infections from 2009-2011. Multiplex RT-PCR was used to screen common respiratory viral pathogens. For HPIV-3-positive specimens, nested RT-PCR was used to amplify the HN gene of HPIV-3. The nucleotides of Hemagglutinin-neuraminidase(HN)gene of 13 HPIV-3 positive strains identified in Gansu and Shaanxi Provinces were successfully sequenced and compared with those downloaded from GenBank. The phylogenetic analysis based on the nucleotides sequence of HN gene showed that 13 HPIV-3 strains belonged to sub-cluster C3 with little sequence variation (overall nucleotide divergence of 0.2%-2.3% and amino acid divergence at 0-1.1%). Compared with the complete gene of HPIV-3 strains from U.S.A., Canada, and Australia, the biggest divergence of the nucleotide and amino acid lovels was 6.0% and 3.4%, respectively. The nucleotide divergence between shaanxi09-2 and shaanxi10-H0091 was 0.9%, while the nucleotide divergence between shaanxi10-H005 and gansull-62110372 was 0.5%, between shaanxi09-2 and BJ/291/09 was 0.6%. However, there was no amino acid divergence among them. It is likely that HPIV-3 virus had been transmitting in Gansu and Shaanxi Provinces for several years. Human parainfluenza virus-3 (HPIV-3) circulated in Gansu and Shaanxi Provinces from 2009 to 2011 belonged to sub-cluster C3.

Objective: To investigate the role of Helicobacter pylori (Hp) and its products cytotoxin-associated protein (Cag A), vacuolating cytotoxin (VacA) in childhood acute idiopathic thrombocytopenic purpura (aITP), to evaluate the effect of Hp on their clinical outcome., Methods: Subjects were enrolled according to case-control design, including 184 aITP children and 154 healthy controls. They were inquired for demographic characteristics, the risk factors regarding Hp infection and ITP through a uniformed questionnaire. Patients with Hp infection were diagnosed by combined detection of serum Hp antibodies and stool antigens. CagA and VacA proteins were tested by ELISA method. In addition, clinical data and follow-up data of aITP children were collected. Non-conditional logistic regression and t test were applied for statistical analysis., Results: (1) The prevalence of Hp infection in aITP children and controls were 41.30% and 35.71%, respectively. No association between Hp infection and children aITP was found with OR of 1.170 (95%CI: 0.7163 - 1.673) after adjusting for confounding variables. (2) No statistical differences regarding initial platelet counts, megakaryocytes counts and the constituent ratio were found between the aITP children with and without Hp infection (P > 0.05). (3) No differences regarding initial platelet counts were found between aITP children with and without the expression of CagA (P > 0.05). The follow-up data showed that 32.88% of aITP children with Hp infection, as well as 29.70% of aITP children without Hp infection developed into cITP. No association between Hp infection and development to cITP was found with adjusted OR 1.171 (95%CI: 0.555 - 2.11 2)., Conclusions: The results didn't suggest that Hp is unlikely to play a role in the onset of childhood aITP, and in the development of aITP to cITP.

Objective: To understand the evolutionary relationship between the C4a evolutionary lineage of human enterovirus 71 (HEV71) strains circulating in mainland of China during 2008-2010 and 2008 Fuyang strains and study the prevalence and transmission patterns of 2008 Fuyang strains., Methods: Download all the complete VP1 ( > or = 891 bp) or approximate complete VP1 (> or = 876 bp) gene nucleotide sequences from GenBank of HEV71 strains circulating in Mainland of China during 2008-2010. And analyze the phylogenetic relationship between Fuyang strains and other provinces' strains using the MEGA software, version 5.0., Results: All of the HEV71 isolates circulating in Mainland of China during 2008-2010 were clustered into evolutionary lineage C4a except for eight strains grouped in the genotype A and one isolate belongs to evolutionary lineage C4b; the homology analysis showed there were 96.5%-100% identity between C4a viruses circulating in mainland China during 2008-2010 and 2008 Fuyang strains, and they were evolved from C4b viruses of 1998. The transmission chains of Fuyang strains were mainly transmitted in Guangdong, Jiangsu, Shanghai, Hunan, Shandong provinces., Conclusion: The predominant viruses circulating in Mainland of China during 2008-2010 were evolutionary lineage C4a of human Enterovirus 71; Fuyang transmission chains mainly distributed in southern of China and the Central China around Anhui provinces.

Objective: To study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A., Methods: Eighteen children with mild hemophilia A were enrolled. DDAVP (0.3 μg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment., Results: In 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment., Conclusions: DDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.