This study aimed to analyse the genetically characterize isolates of Echovirus 11 from Longyan City,Fujian Province,and to reveal their genetic relationships with other isolates from China and abroad. Cerebrospinal fluid specimens from patients diagnosed with viral encephalitis or central nervous system (CNS) infections were collected from Longyan First Hospital between January and December 2011. Seven Echo11 strains were isolated and identified using the RIMV serum panel. The entire VP1 coding regions of four strains were sequenced and typed as Echo11 by an online blast program and,subsequently, phylogenet- ic analyses of the VP1 sequences of these stains and others published on GenBank were conducted. There were 600 nucleotides (nt) in each complete VP1 coding region that encoded 200 amino acids (aa). Among those four Echo11 strains, the sequence identities of nt and aa were 100% and 99%-100% respectively. And phylogenetic analyses indicate belong to subtype DS, the homology compared with DS strain (GU393713) were 93% (nt) and 99% (aa). The sequence identities for the nt and aa were 75%-76% and 90%, respectively, between the current isolates from Longyan and the Gregory prototype strain found in 1953. The sequence identity of nt and aa between the Longyan virus strains and the domestic Shandong strains isolated in 2010 were lower, at 74% and 88%-89%, respectively. However,the highest level of ho- mology was found when the Longyan strains were compared with the Netherlands strain (GU393773) found in 2007 (nt and aa identity: 94%-95% and 98%-99%, respectively). The relatively low levels of similarity between domestic isolates suggest that different transmission routes exist for Echo11 in mainland China.
Adolescent ; Amino Acid Sequence ; Base Sequence ; Child ; Child, Preschool ; Chin ; China ; Encephalitis, Viral ; virology ; Enterovirus B, Human ; chemistry ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics

Objective To analyze the genetic characteristics of echovirus type 25( Echo25) strains isolated from patients with viral encephalitis in Longyan city. Methods Cerebrospinal fluid(CSF) specimens were collected from hospitalized patients with viral encephalitis or central nervous system infection in Longyan,2012. Enteroviruses(EV)were isolated from the specimens and then identified. Four strains of Echo25 were screened out by using serum neutralization test. Coding sequence of the VP1 region of the 4 Echo25 strains were amplified by RT-PCR and then sequenced. A phylogenetic tree was constructed to ana-lyze the nucleotide sequence homology between those sequences and the sequences of reference Echo25 strains available in the GenBank database. Results The VP1 nucleotide sequences of Echo25 strains isola-ted in Longyan were 498 bp in length,encoding 166 amino acid residues. The homology analysis showed that the VP1 nucleotide sequences of 3 strains were identical,sharing 97% homology in nucleotide with the rest strain. The Echo25 strains isolated in Longyan were highly similar to KJ957190( Beijing,2010)and HM031189(Henan,2008)strains. Phylogenetic analysis revealed that the Echo25 strains isolated in Longyan belonged to genotype B1. Conclusion Echo25 was one of the pathogens causing viral encephalitis in Longyan in 2012 and different transmission chains of Echo25 had emerged. This study indicates that it is necessary to strengthen the surveillance for EV and understand the genetic variation of the Echo25 for provi-ding better supportive evidences for the prevention and control of related diseases.

Objective To explore the influence of alpha-fetoprotein (AFP) on the expression of drug-resistance gene MDR1 and chemotherapeutic sensitivity in hepatocellular carcinoma (HCC) cells.Methods A HCC cell line SMMC-7721/AFP, which was stably transfected with AFP gene, was established.mRNA and protein expressions of AFP and MDR1 were detected by real-time PCR and Western Blot,respectively.The sensitivity of SMMC-7721/AFP and SMMC-7721/EGFP cells with or without MDR1 silencing by siRNA to doxorubicin was tested by MTT assay.Immunohistochemistry was used to detect the expression of MDR1 genes-coded protein Pgp in 60 cases of HCC tissues, and the relationship between Pgp expression and serum AFP levels was analyzed.Results AFP mRNA and protein could be detected in SMMC-7721/AFP cells, but not in control cells, indicating that the AFP stably transfected cell line was successfully established.MDR1 mRNA and protein levels were higher in SMMC-7721/AFP cells than those in SMMC-7721/EGFP cells.MDR1 mRNA level in SMMC-7721/AFP cells was (52.7 ± 1.5) times as high as that in SMMC-7721/EGFP cells (P ＜ 0.05).The resistance to doxorubicin was increased by (12.8 ± 1.1) times after AFP transfection (P ＜ 0.05).The chemosensitivity to doxorubicin was increased after the expression of MDR1 was knocked down by siRNA.The expression of Pgp in HCC tissues was positively correlated with the serum AFP levels.Conclusion AFP could induce drug-resistance to doxorubicin in HCC cells by increasing the expression of MDR1.

Objective To analyze the pulse-field gel electrophoresis (PFGE) patterns and antibiotic resistance of Salmonella enteritidisstrains isolated in Longyan city in order to provide reference for effective control, clinical diagnosis and treatment of outbreaks of food-borne diseases.Methods Seventy-seven Salmonella enteritidis strains isolated from food poisoning cases, livestock and poultry meat samples collected for food safety risk monitoring and sporadic cases in 2014 and 2016 were analyzed by PFGE.Antibiotic susceptibility testing was carried out to determine the minimum inhibitory concentrations (MIC) of 16 common antibiotics to 57 strains.Results Seventeen PFGE types were identified among the 77 Salmonella strains including three predominant types (P4, P6 and P8) and were grouped into three major pulsotypes (Cluster1-3) at 90% similarity level.The predominant types causing outbreaks of Salmonella enteritidis infection were P8, P6, P4 and P14, which were responsible for five cases of food poisoning.These isolated strains were highly resistant to nalidixic acid (93%), followed by resistance towards ampicillin (74%) and streptomycin (54%).Forty-one strains were resistant to three or more antibiotics (71.93%).Conclusion The PFGE types of Salmonella enteritidis strains isolated in Longyan city exhibit polymorphism and are clustered into three major pulsotypes.PFGE molecular typing can provide early-warning of sporadic outbreaks and help to identify the sources of infections in Salmonella enteritidis-related food poisoning.Attentions should be paid to multidrug resistant Salmonella enteritidis.Surveillance for multidrug resistant strains and usage of antibiotics should be strengthened.

Objective:To investigate the application value of contrast-enhanced ultrasound combined with serum CXC chemokine ligand 8 (CXCL8) and CXC chemokine receptor 2 (CXCR2) levels detection in the efficacy evaluation of patients with primary liver cancer after transcatheter arterial chemoembolization (TACE) .Methods:A total of 80 patients with primary liver cancer who were diagnosed and treated in Huanggang Central Hospital of Hubei Province from June 2019 to January 2022 were selected as the research objects. The therapeutic efficacy was evaluated 2 months after TACE treatment. According to the pathological diagnosis, the patients were divided into complete inactivation group ( n=30) and residual lesion group ( n=50) . The levels of serum CXCL8 and CXCR2 were measured by enzyme linked immunosorbent assay (ELISA) double antibody sandwich method, and contrast-enhanced ultrasonography was performed on the patients. Receiver operating characteristic (ROC) curve was applied to analyze the value of serum CXCL8 and CXCR2 in evaluating the efficacy of TACE in patients with primary liver cancer. Kappa test was applied to test the consistency of contrast-enhanced ultrasound and contrast-enhanced ultrasound combined with serum CXCL8 and CXCR2 in evaluating the efficacy of TACE in patients with primary liver cancer and the results of pathological diagnosis. Results:Compared with the complete inactivation group, the levels of serum CXCL8 [ (7.12±1.68) ng/ml vs. (5.07±1.25) ng/ml] and CXCR2 [ (3.62±0.79) ng/ml vs. (2.43±0.67) ng/ml] in the residual lesion group were obviously higher ( t=5.79, P<0.001; t=6.89, P<0.001) . The areas under the curve of CXCL8 and CXCR2 in evaluating the efficacy of TACE in patients with primary liver cancer were 0.827 and 0.801 respectively, the specificities were 73.3% and 76.7%, and the sensitivities were 70.0% and 72.0% respectively. The concordance between contrast-enhanced ultrasound and pathological diagnosis was moderate, and the Kappa value was 0.49 ( P<0.001) . The concordance between contrast-enhanced ultrasound combined with serum CXCL8 and CXCR2 and pathological diagnosis was high, and the Kappa value was 0.62 ( P<0.001) . The sensitivity of contrast-enhanced ultrasound combined with serum CXCL8 and CXCR2 in evaluating the efficacy of TACE in patients with primary liver cancer was 90.0%, which was higher than the sensitivity of contrast-enhanced ultrasound (72.0%, χ2=5.26, P=0.022) , CXCL8 (70.0%, χ2=6.25, P=0.012) and CXCR2 (72.0%, χ2=5.26, P=0.022) . Conclusion:Contrast-enhanced ultrasound can detect residual lesions after TACE in patients with primary liver cancer to a certain extent, and its combination with serum CXCL8 and CXCR2 can effectively improve the evaluation efficiency of the efficacy of TACE treatment in patients with primary liver cancer.

OBJECTIVE To investigate the mechanism of Fuke duanhongyin capsule in improving dysfunctional uterine bleeding （DUB） in rats. METHODS Pregnant SD rats were randomly divided into blank control group （distilled water）， model control group （distilled water） and Fuke duanghongyin capsule group （1.296 g/kg）， with 6 rats in each group. Except for blank control group， DUB model was established in other groups. After modeling， they were given relevant medicine/distilled water 10 mL/kg intragastrically， once a day， for consecutive 14 d. After medication， uterine index， ovary index and hemorheology indexes （whole blood high shear relative index， whole blood low shear relative index， erythrocyte aggregation index， Carson viscosity） of rats were all determined. The pathological changes of uterus and ovary tissues were observed； mRNA relative expression levels of epidermal growth factor （EGF）， hypoxia-inducible factor 1α （HIF-1α）， epidermal growth factor receptor （EGFR） and serine/ threonine-protein kinase 1（AKT1）， and protein relative expression levels of EGF and EGFR in uterine tissue were all measured. RESULTS Compared with model control group， the uterine index， ovary index， erythrocyte aggregation index were all decreased significantly in Fuke duanghongyin capsule group （P＜0.05 or P＜0.01）； whole blood high shear relative indexes， mRNA relative expression levels of EGF， HIF-1α and EGFR and protein relative expression levels of EGF and EGFR in uterine tissue were all increased significantly （P＜0.05 or P＜0.01）. The pathological changes of ovary and uterus were improved. CONCLUSIONS Fuke duanhongyin capsule can improve DUB in rat， the mechanism of which may be related to the activation of EGF-EGFR signaling pathway.

ObjectiveTo explore the correlation between gross motor function and spinal range of motion (ROM) in children with bilateral spastic cerebral palsy. MethodsA total of 33 children with bilateral spastic cerebral palsy in Beijing Bo'ai Hospital from December, 2019 to March, 2022 were selected and divided into group 1 (grade Ⅰ, n = 12), group 2 (grade Ⅱ, n = 11) and group 3 (grade Ⅲ, n = 10) according to Gross Motor Function Classification System (GMFCS). SpinalMouse (spine morphometric machine) was used to measure the ROM of forward and backward flexion, and left and right lateral flexion of trunk, respectively, in sitting. Their gross motor ability was then further tested using the Gross Motor Function Measure (GMFM) 88 item. ResultsThe ROM of backward flexion, the left and right lateral flexion, and the total ROM of sagittal and coronal plane were positively correlated with the scores of GMFM88 (r > 0.424, P < 0.05), and were negatively correlated with the grade of GMFCS (|r| > 0.424, P < 0.05). ConclusionSpinal ROM correlates with gross motor function in children with bilateral spastic cerebral palsy.

Traditional Chinese medicines (TCMs) possess a rich historical background, unique theoretical framework, remarkable therapeutic efficacy, and abundant resources. However, the modernization and internationalization of TCMs have faced significant obstacles due to their diverse ingredients and unknown mechanisms. To gain deeper insights into the phytochemicals and ensure the quality control of TCMs, there is an urgent need to enhance analytical techniques. Currently, two-dimensional (2D) chromatography, which incorporates two independent separation mechanisms, demonstrates superior separation capabilities compared to the traditional one-dimensional (1D) separation system when analyzing TCMs samples. Over the past decade, new techniques have been continuously developed to gain actionable insights from complex samples. This review presents the recent advancements in the application of multidimensional chromatography for the quality evaluation of TCMs, encompassing 2D-gas chromatography (GC), 2D-liquid chromatography (LC), as well as emerging three-dimensional (3D)-GC, 3D-LC, and their associated data-processing approaches. These studies highlight the promising potential of multidimensional chromatographic separation for future phytochemical analysis. Nevertheless, the increased separation capability has resulted in higher-order data sets and greater demands for data-processing tools. Considering that multidimensional chromatography is still a relatively nascent research field, further hardware enhancements and the implementation of chemometric methods are necessary to foster its robust development.

Buxue Yimu Pill (BYP) is a classic gynecological medicine in China, which is composed of Angelica sinensis (Oliv.) Diels, Leonurus japonicus Houtt, Astragalus membranaceus (Fisch.) Bunge, Colla corii asini and Citrus reticulata Blanco. It has been widely used in clinical therapy with the function of enriching Blood, nourishing Qi, and removing blood stasis. The current study was designed to determine the bioactive molecules and therapeutic mechanism of BYP against hemorrhagic anemia. Herein, GC-MS and UPLC/Q-TOF-MS/MS were employed to identify the chemical compounds from BYP. The genecards database (https: //www.genecards.org/) was used to obtain the potential target proteins related to hemorrhagic anemia. Autodock/Vina was adopted to evaluate the binding ability of protein receptors and chemical ligands. Gene ontology and KEGG pathway enrichment analysis were conducted using the ClusterProfiler. As a result, a total of 62 candidate molecules were identified and 152 targets related to hemorrhagic anemia were obtained. Furthermore, 34 active molecules and 140 targets were obtained through the virtual screening experiment. The data of molecular-target (M-T), target-pathway (T-P), and molecular-target-pathway (M-T-P) network suggested that 32 active molecules enhanced hematopoiesis and activated the immune system by regulating 57 important targets. Pharmacological experiments showed that BYP significantly increased the counts of RBC, HGB, and HCT, and significantly down-regulated the expression of EPO, IL-6, CSF3, NOS2, VEGFA, PDGFRB, and TGFB1. The results also showed that leonurine, leonuriside B, leosibiricin, ononin, rutin, astragaloside I, riligustilide and levistolide A, were the active molecules closely related to enriching Blood. In conclusion, based on molecular docking, network pharmacology and validation experiment results, the enriching blood effect of BYP on hemorrhagic anemia may be associated with hematopoiesis, anti-inflammation, and immunity enhancement.
Anemia/drug therapy* ; Drugs, Chinese Herbal ; Humans ; Molecular Docking Simulation ; Network Pharmacology ; Tandem Mass Spectrometry