Objective:To study the epidemiological and genetic characteristics of coxsackievirus B3 (CVB3) circulating in Guangdong Province from 2008 to 2021.Methods:This study collected the specimens of hand, foot, and mouth disease (HFMD) cases from 2008 to 2021 that were positive for other enteroviruses except for enterovirus 71 (EV71), coxsackievirus A16 (CVA16), and CVA6, as well as the specimens of herpangina and neonatal infection cases from 2020 to 2021. Enteroviruses in these specimens were detected and their types were identified. CVB3 strains were isolated and the entire VP1 sequences of CVB3 strains were amplified and sequenced. The genetic features of CVB3 strains were analyzed using DNAStar 7.1 and MEGA 6.06 software packages.Results:Among 3 484 HFMD cases positive for other enteroviruses from 2008 to 2021, CVB3-positive cases accounted for 1.6% (57/3 484); among 560 cases of herpangina from 2020 to 2021, CVB3-positive cases accounted for 2.1% (12/560); one neonatal infection case in 2021 was positive for CVB3. CVB3-positive cases accounted for 67.1% (47/70) in 2021 and 18.6% (13/70) in 2020, while there were less than five cases in other years. Forty-eight CVB3 strains were isolated and the entire VP1 sequences of 26 CVB3 strains were obtained. Phylogenetic analysis indicated that the CVB3 strains could be divided into eight genotypes (A-H) and the strains of genotypes A, D and E were prevalent in the Chinese mainland. The 26 CVB3 strains isolated in Guangdong Province shared 80.2%-100.0% nucleotide homology, and belonged to two genotypes of D and E, with genotype D prevalent from 2008 to 2017 and genotype E prevalent from 2020 to 2021.Conclusions:CVB3 is prevalent sporadically in Guangdong Province from 2008 to 2017, but the epidemic intensity increased during 2020 and 2021. CVB3 strains of genotypes D and E are prevalent in Guangdong Province during 2008 to 2021, with genotype E being the prevalent genotype during 2020 and 2021.

Objective:To observe the therapeutic effect of echinacoside (ECH) on liver injury and glucose metabolism disorder in sepsis rats induced by cecal ligation and puncture (CLP), and to explore its possible mechanism.Methods:Forty eight male Sprague Dawley (SD) rats were randomly divided into four groups: sham group (sham), model group (CLP), treatment group (CLP+ ECH) and inhibitor group (CLP+ ECH+ EX527). The sham group only received laparotomy, and the model group underwent CLP. The treatment group was intragastric administration of echinacea (30 mg/kg) every day after CLP modeling. The inhibitor group was injected with silence information regulator 1 (SIRT1) inhibitor EX527 (5 mg/kg) one hour before CLP, and then treated the same as the treatment group. Fasting blood glucose, insulin and serum biochemical indexes were detected in virous groups. The serum levels of interleukin (IL)-1 β, IL-6 and tumor necrosis factor-α(TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). 2′, 7′- dichlorofluorescein diacetate (DCFH-DA) staining was used to observe the production of reactive oxygen species (ROS) in liver tissue of rats in each group; hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissue in each group; The expressions of SIRT1, glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), phosphorylated signal transducers and activators of transcription 3 (p-STAT3) and phosphorylated protein ki-nase B(p-AKT) were detected by Western blot.Results:Compared with sham group, the levels of serum glucose, serum insulin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), ROS, IL-1β, IL-6 and TNF-α in model group increased, while the liver glycogen and survival rate decreased (all P<0.05). After echinacoside treatment, the serum glucose, serum insulin, ALT, AST, ROS , IL-1β, IL-6 and TNF-α levels decreased, and the liver glycogen and survival rate increased (all P<0.05); After SIRT1 inhibitor intervention, the levels of serum insulin, ALT, AST, IL-6 and ROS in the inhibitor group increased ( P<0.05). HE staining showed that there were infiltration and necrosis of inflammatory cells in the liver tissue of model group, and echinacoside could significantly reduce the focal and massive necrosis; Western blot showed that compared with the sham group, the expression levels of SIRT1, p-STAT3 and p-AKT protein in the model group decreased, while the expression levels of G6Pase and PEPCK protein increased ( P<0.05); After echinacoside treatment, the expression levels of SIRT1, p-STAT3 and p-AKT increased, while the expression levels of G6Pase and PEPCK decreased ( P<0.05). After SIRT1 inhibitor intervention, the expression of SIRT1, p-STAT3 and p-AKT protein decreased, and the expression of G6Pase and PEPCK protein increased in the inhibitor group ( P<0.05). Conclusions:Echinacoside is a potential therapeutic agent for sepsis associated liver injury and glucose metabolism disorders, which may play a role by targeting SIRT1 to activate STAT3 and AKT in the liver.

Objective:To investigate whether miRNA-296-5p can inhibit enterovirus 71 (EV71) virus replication in neural cells SK-N-SH by targeting the phosphatase and tensin homology deleted on chromosome 10 (PTEN)/phosphatidylinositol 3-kinase (PI3K)/serine/threonine kinases (AKT) signaling pathway.Methods:Serum samples were collected from patients with EV71 virus-infected hand-foot-mouth disease and normal physical examination, and the expressions of serum inflammatory factors procalcitonin (PCT), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6), IL-1β, and IL-13 were detected by enzymelinked immunosorbent assay (ELISA). Human neuroblastoma SK-N-SH cells infected by EV71 virus in logarithmic growth phase were set up as control group, miRNA-296-5p mimic group and miRNA-296-5p inhibitor group. The transfection was carried out according to the Lipofectamine 2000tm cell transfection reagent. The expression of EV71-VP1 gene mRNA and protein and PTEN/PI3K/Akt signal pathway related molecules in three groups of cells was observed by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blot.Results:ELISA test results showed that the levels of serum inflammatory factors PCT, CRP, TNF-α, IL-1β, IL-6 and IL-13 in patients with EV71 virus-infected hand, foot and mouth disease were significantly higher than those in normal physical examination ( P<0.05). The results of qRT-PCR and Western blot showed that the mRNA and protein levels of mirna-296-5p and PTEN in SK-N-SH were significantly decreased after EV71 virus infection, while the mRNA and protein levels of EV71-VP1 and molecules related to PI3K/AKT signaling pathway were significantly increased ( P<0.05). The expression of PTEN was significantly increased in the miRNA-296-5p mimic group, and the expression of EV71-VP1 and the activation of the PI3K/AKT signaling pathway were inhibited, while the effect was reversed in the miRNA-296-5p inhibitor group ( P<0.05). Conclusions:MiRNA-296-5p inhibits the replication of EV71 virus in neural cells SK-N-SH by targeting the PTEN/PI3K/AKT signaling pathway, while reducing the cellular inflammatory response, targeting miRNA-296-5p and downstream PTEN/PI3K/AKT The signal pathway is expected to provide therapeutic targets and theoretical basis for the treatment of hand-foot-mouth disease caused by clinical EV71 virus.

Objective: To analyze and discuss the clinical value of single photon emission computed tomography(SPECT)/CT fusion imaging in differentiating benign and malignant spinal lesions. Methods: From January 2017 to January 2018, 70 cases with benign and malignant spinal lesions in Shanxida Hospital were studied.SPECT/CT fusion imaging and SPECT examination were used to compare the diagnosis results between the two groups. Results: After SPECT diagnosis, a total of 98 bone metastatic lesions were detected, including 10 false positive lesions, 60 benign lesions and 28 false negative lesions.After SPECT/CT fusion imaging, there were 100 lesions, including 4 false positive lesions, 59 benign lesions and 37 false negative lesions.In this study, a total of 145 lesions were found in 70 patients with benign and malignant spinal lesions, including 95 benign lesions and 50 malignant lesions.The specificity, sensitivity and accuracy of SPECT/CT fusion imaging [95.79%(91/95), 92.00%(46/50), 93.79%(136/145)] were higher than those of the control group [82.11%(78/95), 64.00%(32/50), and 82.76%(120/145)], the differences were statistically significant(χ²=9.048, 11.422, 9.578, all P<0.05). Conclusion In the diagnosis of benign and malignant spinal diseases, SPECT/CT fusion imaging can accurately distinguish benign and malignant lesions, and clearly reflect the development of the disease, which lay a solid foundation for good treatment in the future.

Objective: To analyze the clinical features of acute liver failure in critically ill children, to investigate the risk factors affecting the prognosis of acute liver failure in critically ill children, and to provide evidence for clinical diagnosis and prognosis. Methods: The data of 85 children with acute liver failure who conformed to the diagnostic criteria from January 2011 to January 2017 at Hunan Children′s Hospital were analyzed.According to the prognosis of children with acute liver failure, all children were divided into the survival group (49 cases) and the death group (36 cases). Univariate and multivariate Logistic regression methods were used to analyze the risk factors affecting the prognosis of children with acute liver failure. Results: There were statistically significant differences of enzyme bile isolation[26.5% (13/49 cases) vs.55.6%(20/36 cases), χ²=7.361, P=0.007], hepatic encephalopathy [16.3%(8/49 cases) vs.38.9%(14/36 cases), χ²=5.507, P=0.019], total bilirubin [72.30(3.93-428.80) μmol/L vs.153.08(3.23-776.26) μmol/L, U=-2.283, P=0.027], albumin [35.02(22.89-45.30) g/L vs.28.90(18.30-40.26) g/L, U=4.640, P=0.000], alanine aminotransferase [1 626.10(23.01-9 518.41) U/L vs.533.08(7.02-5 163.83) U/L, U=2.992, P=0.004], aspartate aminotransferase [1 611.20(34.51-15 850.02) U/L vs.715.54(60.06-10 280.42) U/L, U=2.312, P=0.023], blood ammonia [71.02(16.04-148.56) μmol/L vs.81.02(33.04-274.02) μmol/L, U=-2.057, P=0.046], prothrombin time [23.10(13.61-81.23) s vs.33.91(12.62-84.57) s, U=-2.364, P=0.022], activated partial prothrombin time [52.91(4.02-181.02) s vs.67.35(31.20-180.02) s, U=-2.226, P=0.029], and end-stage liver disease model score [13.00(-9.00-52.00) score vs.27.50(-9.00-88.00) score, U=-3.082, P=0.003] in the children with acute liver failure between the survival group and the death group.Multivariate Logistic regression analysis showed that total bilirubin (OR=0.236, 95%CI: 0.059-0.946, P=0.041), albumin(OR=0.854, 95%CI: 0.746-0.976, P=0.019), enzyme bile separation (OR=1.063, 95%CI: 1.031-1.119, P=0.005) and hepatic encephalopathy (OR=1.439, 95%CI: 1.021-2.043, P=0.017) were risk factors for the prognosis of children with acute liver failure. Conclusions The level of the total bilirubin and the ratio of enzyme bile isolation and hepatic encephalopathy are positively correlated but the level of albumin is negatively correlated with the prognosis of critically ill children with acute liver failure.

To analyze the clinical manifestations and gene mutations in children with congenital insensitivity to pain with anhidrosis (CIPA), and review related literature. An infant diagnosed with congenital insensitivity to pain with anhidrosis was reported. The main clinical manifestations of the infant were painless, no sweat, and repeated fever. Peripheral blood of the infant and his parents was collected, and candidate variants were confirmed by Sanger sequencing. The results of molecular genetic analysis showed that there were compound heterozygous mutations (c.36G>A, c.851-33T>A) of neurotrophic tyrosine kinase receptor type 1 (NTRK1) in the infant. c.36G>A and c.851-33T>A were inherited from his father and mother, respectively. c.851-33T>A is a previously reported mutation, c.36G>A is an unreported mutation, which can lead to the tryptophan changing into a stop codon. According to the American College of Medical Genetics and Genomics (ACMG) variant interpretation guidelines, the mutation is interpreted as pathogenic, and the biological hazard is potentially harmful. Congenital insensitivity to pain with anhidrosis is a rare inherited disorder. Genetic molecular genetic analysis is helpful to diagnose and discover new gene mutations.
Channelopathies ; Humans ; Infant ; Mutation ; Pain Insensitivity, Congenital ; Receptor, trkA

Objective: To compare the clinical features of papillary thyroid microcarcinoma (PTMC) and non-microcarcinoma papillary thyroid carcinoma (PTC). Methods: Clinical data of 522 patients (156 males, 366 females, age: 16-77 years) with PTC treated with ¹³¹I from January 2014 to September 2018 were analyzed retrospectively. Patients were divided into PTMC group (46 males, 139 females, age: (38.5±6.5) years; tumor diameter≤1.0 cm) and non-microcarcinoma PTC group (110 males, 227 females, age: (40.5±4.5) years; tumor diameter>1.0 cm). General information and TNM status of patients were compared. The t′ test and χ² test were used to analyze the data. Results: Patients was relatively young in PTMC group (t′=2.20, P<0.05), but no difference was observed in gender between 2 groups (χ²=3.45, P>0.05; mostly females). The incidence of extraglandular invasion in PTMC group was significantly lower than that in PTC group (33.51%(62/185) vs 56.08%(189/337); χ²=24.37, P<0.01), mainly in peripheral muscle/fibrous adipose tissue (χ²=11.01, P<0.01) and tracheal infiltration (χ²=5.35, P<0.05). Nodular goiter and Hashimoto′s thyroiditis were commonly shown in both groups, and the tumor distribution was bilobar and/or multi-foci. The rate of regional lymph node metastasis was higher in non-microcarcinoma PTC group (88.43% (298/337) vs 82.16% (152/185); χ²=3.94, P<0.05), and central lymph node metastasis occurred more in PTMC group (χ²=5.75, P<0.05). Besides, non-microcarcinoma PTC group was likely to involve more lymph node areas (χ²=5.69, P<0.05) and distant metastasis (9.50% (32/337) vs 2.16% (4/185); χ²=10.00, P<0.01). There were no differences of extraglandular infiltration, tumor distribution or lymph node metastasis between moderate- and high-risk PTMC and non-microcarcinoma PTC groups (χ² values: 0.01-3.33, all P>0.05). Conclusions Clinical characteristics of PTC can be influenced by tumor diameter. The features of primary tumor and lymph node metastasis between patients with moderate- and high-risk PTMC and non-microcarcinoma PTC patients are similar, which suggests that ¹³¹I therapy is necessary to patients with moderate- and high-risk PTMC.

Objective To compare the diagnostic value of ventilation/perfusion ( V/Q) combined with pulmonary perfusion single photon emission computed tomography combined with CT ( SPECT/CT) fu-sion tomography imaging and computed tomographic pulmonary angiography ( CTPA) in evaluation of pulmo-nary embolism. Methods We retrospectively analyzed 60 patients with clinically suspected pulmonary em-bolism diagnosed in Shanxi Dayi Hospital from May 2015 to May 2017. All patients underwent pulmonary V/Q imaging and lung perfusion SPECT/CT fusion tomography, and CTPA inspections were completed with-in 3 days. The final clinical diagnosis and follow-up confirmed the presence or absence of pulmonary embol-ism. The diagnostic efficacy of two imaging methods for pulmonary embolism were calculated and compared. Results Of the 60 cases of patients, 33 cases were diagnosed with pulmonary embolism; the sensitivity, specificity, and accuracy of V/Q combined with pulmonary perfusion SPECT/CT fusion tomography were 96. 97％ (32/33), 92. 59％ (25/27) and 95％ (57/60), respectively; the sensitivity, specificity, and accuracy of CTPA were 81. 82％ (27/33), 92. 59％ (25/27) and 86. 67％ (52/60), respectively; both have no diagnostic uncertainty, there was no significant difference in the qualitative diagnosis of pulmonary embolism between the two examination methods ( P >0. 05 ) . V/Q combined with pulmonary perfusion SPECT/CT fusion tomography found 253 lung segment and 50 unmatched sub-pulmonary segments, including 15 V/Q mismatch lung segment and 5 sub-segment caused by lung lesions which were confirmed by lung perfusion SPECT/CT fusion image ( 5 interlobular or pleural effusion, 4 local emphysema and pulmonary bulla, 3 interlobular hypertrophy, 8 pulmonary parenchymal inflammation); CTPA found 3 sub-segmental pulmonary filling defects, 6 cases of false-negative cases were multiple sub-pulmonary segment pulmonary embolism. Conclusions V/Q combined with pulmonary perfusion SPECT/CT fusion tomography is similar to CTPA in diagnosing pulmonary embolism, and both of them have better diagnostic efficacy; the former has advantages in the diagnosis of sub-pulmonary segment pulmonary embolism, and can exclude false-posi-tive diagnoses due to other lung lesions and provide additional diagnostic information for lung disease.

Objective To investigate the mechanism of miR-758 in hepatocellular carcinoma cell HepG2,and to investigate the regulatory role of miR-758 on astrocyte elevated gene-1 (AEG-1).Methods Transient transfection of miR-758 into HepG2 cells was performed to study the effect of miR-758 on tumor cell metastasis by transwell migration and invasion experiments.CCK8 assay was used to detect the cell proliferation activity.The cell cycle was analyzed by flow cytometry.The effect of miR-758 on epidermal mesenchymal transition (EMT) was determined by the expression of EMT markers.Transient transfection of miR-758 into human umbilical vein endothelial cells (HUVECs) was performed to explore the effect of miR-758 on luminal formation.AEG-1 3'UTR containing the binding site of miR-758 was constructed into luciferase expression vector.The miR-758 and the vector was co-transfected into HepG2 cells.And then the change in expression level of AEG-1 protein was detected through Western Blot.Results The overexpression of miR-758 inhibited HepG2 cell migration and invasion,as well as the cell proliferation and the cell cycle.The miR-758 was also found to inhibit EMT of HepG2 cells and the lumen formation of HUVEC cells.After the co-transfection of miR-758 with the plasmid containing AEG-1 gene 3'UTR into HepG2 cells,the luciferase expression was decreased.The luciferase expression was restored when the binding site of miR-758 in the 3'UTR was mutated.Further evidence by Western Blot showed the protein level of AEG-1 in HepG2 cells was significantly decreased after transfection of miR-758.Conclusions The miR-758 negatively regulates multiple steps during cancer metastasis,including cell migration,invasion,cell proliferation,EMT,as well as angiogenesis.And AEG-1 has been identified as a downstream target of miR-758.

Objective To investigate the role of neuroglobin ( NGB) in oxygen-glucose deprivation and reoxygenation ( OGD/R ) induced mitochondrial depolarization and reactive oxygen species ( ROS ) production in a human neuroblastoma cell line (SH-SY5Y).Methods SH-SY5Y cells were transfected with lentivirus to establish a stable cell line of NGB knockdown ( KD).After treated with OGD/R, cells were collected at different time points to analyze NGB mRNA and protein levels.Furthermore, cells were stained with JC-1 and DCFH-DA to evaluate mitochondrial depolarization and ROS production by inverted fluorescence microscope.Also, to determine the neurotoxicity , we measured the lactate dehydrogenase ( LDH) level in the cell culture medium.Results After the treatment of OGD/R, the NGB mRNA and protein started to elevate and peak at 4 h and 8 h (2.04 ±0.35 fold,1.69 ±0.18 fold).Compared with the vector group , NGB KD group had much more mitochondrial depolarization [ JC-1 red/green ( 1.10 ±0.10 ) vs (1.46 ±0.11),P<0.05] and ROS production [DCFH-DA fluorescence (36.30 ±5.32) vs (16.26 ± 2.97),P<0.05].Furthermore, NGB KD groups had a higher level of LDH release [(63.42 ±6.14)%vs (49.65 ±5.09 )%, P <0.05 ].Conclusions NGB plays an important role in the homeostasis of mitochondria.Knockdown of NGB results in increased mitochondrial depolarization , ROS production and neurotoxicity under hypoxia circumstances.