Objective To summarize the role of nuclear factor kappa B(NF-?B) in the occurrence and progression of various sorts of liver injury.Methods Literatures on the structures,property of molecular biology and function of NF-?B,as well as its relationships with liver injury were collected and reviewed.Results NF-?B was an important nuclear factor existed in cells widely distributed in most cell types.The activation of NF-?B was induced by various sorts of liver injury.The activated NF-?B could affect the liver injury by regulating cytokines,adhesion molecules,and activating factor involving in immunologic reaction,inflammatory reaction and the apoptosis.Conclusion NF-?B plays an important role during the occurrence and progression of liver injury,and may become a new target in the treatment of liver injury.

ObjectiveTo identify the pathogen of an aseptic meningitis outbreak which occurred in Linyi City of Shandong Province during the summer of 2009,and to analyze the genetic variations of Coxsackicvirus B5 (CVB5) isolates.MethodsForty-two cerebrospinal fluids (CSF) specimens were collected from aseptic meningitis cases and virus isolation was performed. The viral RNA was extracted and amplified from the positive specimens using reverse transcription polymerase chain reaction (RT-PCR).The partial VP1 coding region was purified and sequenced. The phylogenetic trees based on VP1 sequences were constructed among CVB5 isolates and those in GenBank.ResultsSeventeen enteroviruse strains were isolated from 42 CSF samples with 40.5％ isolation positive rate. All these strains were identified as CVB5 using both microneutralization test and molecular typing methods. Homology comparisons indicated that the nucleotide acid identities and amino acid sequence identities were 92.3 ％- 100.0％ and 98.7 ％- 100.0％,respectively among these CVB5 isolate.s,and compared with the Faulkner prototype strain,which were 81.0％-82.4％ and 96.6％97.0％,respectively.Phylogenetic analysis on VP1 sequences showed that all CVB5 could be separated into four genogroups of A,B,C and D.Isolates of this outbreak belonged to genogroup D.Interestingly,two distinct genogroups in the phylogenetic tree were observed among the 17 isolates.Conclusions CVB5 is responsible for the outbreak of aseptic meningitis in Linyi City of Shandong Province,China. The genetic diversity is high among the isolates and all belong to genogroup D.

Objective To analyze the genetic characteristics of echovirus 6 ( E-6) strains isolated from patients with acute meningitis/encephalitis syndrome ( AMES) in 2014 and sewage samples in 2013—2014 in Shandong province and to investigate their correlations.Methods Enterovirus strains were isolated from cerebrospinal fluid, stool and throat swab samples collected from 940 cases of AMES and 96 sewage samples used for environmental surveillance.The positive isolates were identified by molecular typing meth-od.Homologous and phylogenetic analyses based on the VP1 sequences of E-6 isolates were performed.Re-sults Altogether 47 E-6 strains were isolated from patients with AMES in 2014, accounting for 29.56%of all isolated enteroviruses ( EVs) strains.No E-6 strains were isolated from AMES cases in 2013.Data of the environmental surveillance showed that E-6 virus strains had been frequently detected in sewage samples since the summer of 2013 to the end of 2014.In total, 40 E-6 virus strains were isolated (7.87% of total isolated EVs strains) in 2013 and 139 E-6 virus strains (26.18%) in 2014.Phylogenetic analysis indicated that the E-6 isolates recruited in this study belonged to clusters A and C with high intracluster sequence iden-tities between AMES and environmental isolates.The nucleotide identities were 98.3%-100% among cluster A E-6 virus strains isolated from AMES cases in 2014 and 96.6%-100% among cluster A E-6 virus environ-mental isolates during the surveillance year 2013—2014.The cluster A E-6 virus strains shared 97.1%-100% nucleotide identities between the AMES and environmental isolates.For cluster C E-6 virus strains, the nucleotide identities were 100%, 98.7%-100% and 99.1%-100%, respectively.Conclusion The epidemic of viral encephalitis in Shandong province in 2014 was associated the transmission of two lineages of E-6 virus.Environmental surveillance revealed the potential epidemic of E-6 virus even before the epidemic of viral encephalitis in Shandong province, indicating the possibility of using environmental surveillance for early warning of related diseases.

Objective To analyze the expression characteristics of excision repair cross-complementing 1 (ERCC1), topoisomeraseⅡ (TOPOⅡ), ribonucleotide reductase M1 (RRM1), β3-tubulin and thymidylate synthase (TS) in lung cancer and their associations with the pathological types. Methods The immunohistochemical EnVision method was used to determine the expression of ERCC1, TOPOⅡ, RRM1,β3-tubulin and TS in 548 patients who were diagnosed as lung cancer from January 2011 to December 2014. Variance analysis was performed to analyze their expression characteristics among different pathological types and correlation. Results The expression positive rates of ERCC1, TOPOⅡ, RRM1, β3-tubulin and TS were 61.86 % (339/548), 91.06 % (499/548), 62.59 % (343/548), 73.18 % (401/548) and 70.44 % (386/548), respectively. The expression of ERCC1 was weak positive mostly (P<0.05), meanwhile the expression of TOPOⅡ was medium-strong positive mostly (P<0.05). In ERCC1 group, the positive rate of squamous cell carcinoma was higher than that of adenocarcinoma [57.39 % (167/291) vs. 42.61 % (124/291), P=0.000]. In weak positive of TOPOⅡ group, the proportion of adenocarcinoma was higher than that of squamous cell carcinoma [23.58 % (100/137) vs. 8.73 % (37/137), P=0.000]. In medium-strong positive of TOPOⅡ group, the proportion of squamous cell carcinoma was higher than that of adenocarcinoma [47.41 % (201/287/) vs. 20.28%(86/287), P=0.000]. The expressions of ERCC1, TOPOⅡ, RRM1,β3-tubulin and TS were irrelevant (r=0.4, P=0.397). Conclusions The expressions of ERCC1 and TOPOⅡ are higher in squamous cell carcinoma than those in adenocarcinoma. The expression of ERCC1 is weak positive mostly, meanwhile the expression of TOPOⅡis medium-strong positive mostly. There is no correlation between them.

Objective To explore the effect of Drotaverine hydrochloride on preventing bladder spasm after transurethral prostatectomy.Methods 124 patients after transurethral prostatectomy were divided into patient-controlled epidural analgesia pump group (group I, n=61) and Drotaverine hydrochloride group (group II, n=63). Group I received bupivacaine by patient-controlled epidural analgesia, and the pump was withdrawed after 72 h. Group II received Drotaverine hydrochloride by intramuscular injection, 80 mg every 12 h, and then orally taken after anal exhaust for 3 days. Bladder spasm and adverse reaction were recorded in both groups. Results There was no significant difference in bladder spasm between group I (11.48%) and group II (12.70%) (P>0.05), as well as in side reaction between group I (16.39%) and group II (17.46%) (P>0.05). Conclusion Drotaverine hydrochloride is effective on preventing bladder spasm after transurethral resection of the prostate, with small side effect.

To analyze the genetic characteristics of a polio-I highly variant vaccine recombinant virus in Shandong Province (China) in 2011 and to identify isolates from healthy contacts, two stool specimens from one patient with acute flaccid paralysis (AFP) and 40 stool specimens from his contacts were collected for virus isolation. The complete genome of poliovirus and VP1 coding region of the non-polio enterovirus were sequenced. Homologous comparison and phylogenetic analyses based on VP1 sequences were undertaken among coxsackievirus (CV) B1, CV-B3 isolates, and those in GenBank. One poliovirus (P1/11186), CV-A4 and CV-A8 were isolated from the AFP patient; one CV-A2, Echovirus 3 (E-3), E-12 and E-14, ten CV-B1, and five CV-B3 strains were isolated from his contacts. These results led us to believe that there may be a human enterovirus epidemic in this area, and that surveillance must be enhanced. P1/11186 was a type-1 vaccine-related poliovirus; it combined with type-2 and type-3 polioviruses in 2A and 3A regions, respectively. There were 25 nucleotide mutations with 9 amino-acid alterations in the entire genome. There were 8 nucleotide mutations with 5 amino-acid alterations in the VP1 region compared with the corresponding Sabin strains. Homology analyses suggested that the ten CV-B1 isolates had 97.0%-100% nucleotide and 98.9%-100% amino-acid identities with each other, as well as 92.6%-100% nucleotide and 99.2%-100% amino-acid identities among the five CV-B3 isolates. Phylogenetic analyses on the complete sequences of VP1 among CV-B1 and CV-B3 isolates showed that Shandong strains, together with strains from other provinces in China, had a close relationship and belonged to the same group.
Base Sequence ; Capsid Proteins ; genetics ; immunology ; Child, Preschool ; China ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Poliomyelitis ; etiology ; prevention & control ; virology ; Poliovirus ; classification ; genetics ; immunology ; isolation & purification ; Poliovirus Vaccines ; adverse effects ; genetics ; immunology

Objective:To explore newer enteroviruses (EVs) circulating in China and research their genetic characterization.Methods:Sewage samples were collected in Jinan in January and July of 2018, respectively. PCR amplification of the P1 coding region of EV and next-generation sequencing (NGS) were conducted. Homologous and phylogenetic analyses were performed on the newly identified newer EVs.Results:The EV-A89 (n=2) and EV-C96 (n=1) nucleic acid sequences were detected in the sewage in January and July, respectively and obtained complete P1 coding sequence. The two EV-A89 sequences had 6.1% nucleotide divergence among themselves, and had 88.2%-95.3% homologies with other strains. No close genetic relationship was obtained with the sequences from Xinjiang, China and those from other countries. The EV-C96 sequence in this study had 76.2%-89.4% nucleotide similarities with other isolates throughout the world. Phylogenetic analysis showed that global EV-C96 strains were divided into 5 branches, i. e, A to E. Except for the Shandong 1991 isolate, the other Shandong and domestic strains were all located in branch D.Conclusions:This study added the molecular epidemiologic data of newer enteroviruses EV-A89 and EV-C96, and provides basic data for future research on the epidemic trend of EV-A89 and EV-C96 as well as their association with diseases.

Objective:To explore the effects of deleted in lung and esophageal cancer 1 (DLEC1) on osteosarcoma cells and the underlying mechanism.Methods:Immunohistochemical staining for DLEC1 was scored in sixteen paired osteosarcoma tissues and adjacent normal tissues obtained. The present study was conducted on human osteosarcoma 143B cells which were randomly divided into two groups, pDC316-DLEC1 transfection group and pDC316-Null transfection group. Differences in the proportion of EdU-positive cells, cell cycle distribution, proportion of apoptosis cells, number of migrating and invasive cells, expression of epithelial-mesenchymal transformation (EMT) markers (E-cadherin and vimentin), relative protein expression levels of NF-κB, AKT and ERK signaling pathways were assessed between the pDC316-DLEC1 and pDC316-Null transfection groups in in vitro study. The subcutaneous inoculation model and tail vein injection model were developed to evaluate the differences in subcutaneous tumor volume, subcutaneous tumor weight and pulmonary tumor nodules between the above two groups in in vivo study.Results:The DLEC1 immunostaining scores for osteosarcoma tissues and adjacent normal tissues were 2.88±1.15 and 4.25±1.06, respectively. The proportions of EdU-positive cells (36.47%±1.90% vs 51.47%±2.89%) and S phase cells (33.31%±0.61 vs 43.77%±1.47%) were decreased, while G0/G1 phase cells (46.87%±0.73% vs 35.47%±1.14%) and apoptotic cells (13.83%±1.01% vs 3.30%±0.26%) were increased in the pDC316-DLEC1 transfection group compared to those in the pDC316-Null transfection group. Decreased number of migrating cells (199.00±12.53 vs 369.67±10.02) and invasive cells (104.67±9.07 vs 299.67±12.06) and relative expression of vimentin mRNA (0.59±0.02 vs 1.00±0.02) and protein (0.54±0.08 vs 1.00±0.00) were observed in the pDC316-DLEC1 transfection group, while relative expression of E-cadherin mRNA (2.40±0.05 vs 1.00±0.02) and protein(1.98±0.10 vs 1.00±0.00) in the pDC316-DLEC1 transfection group were higher than those in the pDC316-Null transfection group. The relative protein expression of NF-κB (p65), p-AKT (Ser473) and p-ERK (Thr202/Tyr204) in the pDC316-DLEC1 transfection group were decreased by 51.67%±4.04%, 64.67%±5.51% and 48.67%±4.73% compared to those in the pDC316-Null transfection group. In in vivo study, 143B cells in the pDC316-DLEC1 transfection group formed smaller (320.00±145.22 mm 3vs 798.00±221.94 mm 3) and lighter (0.49±0.17 g vs 0.88±0.14 g) subcutaneous tumors and less metastatic lung nodules (7.71±1.80 vs 20.86±3.53) compared with those in the pDC316-Null transfection group. Conclusion:Overexpression of DLEC1 could suppress the NF-κB/AKT/ERK signaling pathways in 143B cells, which further induces G0/G1 arrest and apoptosis that ultimately inhibits cell proliferation and reduces the metastatic potential through reversing EMT.

Objective:To investigate the complicated virus infection of infants with pertussis and its effect on the disease.Methods:From January 2019 to March 2020, a total of 100 hospitalized infants with pertussis were admitted to the Second Affiliated Hospital of Medical College of Shantou University, nasopharyngeal swabs were collected for detection of ten pathogens including pertussis, namely respiratory syncytial virus(RSV), parainfluenza virus(PIV), bordetella pertussis (BP), human rhinovirus(HRV), human bocavirus(HBoV), human metapneumovirus(hMPV), influenza B virus (INF-B), adenovirus, influenza A virus and cytomegalovirus(CMV). According to the results of pathogen detection, all infants were divided into single detection group of BP(single detection group) and co-detection group of BP combined with viruses(co-detection group). The clinical data of the two groups were retrospectively analyzed and compared to explore the differences of clinical characteristics and its impact on the course of disease.Results:Among 100 cases, there were 54(54.0%) boys and 46(46.0%)girls.The age ranged from 28 days to 2 years and 5 months, with a median age of 3.5 months.Fifty-six cases were classified as single detection group, while 44 cases were included into co-detection group.Among infants in co-detection group, fourteen cases were co-infected with CMV(31.8%, 14/44), seven cases with HRV(15.9%, 7/44), seven cases with PIV(15.9%, 7/44), four cases with RSV(9.1%, 4/44), one case with hMPV(2.2%, 1/44), eight cases with CMV+ HRV(18.2%, 8/44), one case with HRV+ HBoV (2.2%, 1/44), one case with CMV+ PIV(2.2%, 1/44)and one case with CMV+ PIV+ INF-B(2.2%, 1/44). The number of infants in the single detection group who had cyanosis before treatment, requiring repiratory support, PICU admission, severe pneumonia or abnormal myocardial enzymes were higher than those in the co-detection group( P<0.05), while the months of age were lower than that in the co-detection group( P<0.05). When comparing the clinical characteristics of infants over three months of age, only the number of cases of combined cyanosis before treatment and the number of days in hospital were higher in the single detection group than those in the co-detection group ( P<0.05), no statistically significant differences were found in the other clinical characteristics between the two groups( P>0.05). Conclusion:The cases of infants requiring repiratory support, complicated with severe pneumonia or abnormal myocardial enzymes in the single detection group are higher than those in the co-detection group, which may be attributed to the small age of months.

Objective:To study the complete genome characterization of Human Astrovirus (HAstV) in Shandong Province.Methods:Stool samples from acute flaccid paralysis (AFP) surveillance in Shandong Province from 2020 to 2022 were collected, and HAstV nucleic acid was examined by real-time quantitative PCR (qPCR). Next-generation sequencing (NGS) was conducted for the positive samples to obtain complete genome sequences and identify the genotype. Homology comparison and phylogenetic analysis were performed by using BioEdit and Mega software.Results:A total of 667 samples were examined by qPCR, of which 14 were HAstV-positive (2.1%), including HAstV-1 ( n=6), MLB1 ( n=6), MLB2 ( n=1), and VA2 ( n=1). The complete genome sequences were obtained from 11 samples. The six HAstV-1 sequences of this study had 98.2% to 99.9% nt similarities with each other and 87.6% to 98.6% with those from other regions. The four MLB1 sequences of this study had 99.1% to 99.9% nt similarities with each other and 92.2% to 99.4% with those from other regions. The VA2 sequence of this study had 96.0% to 96.3% nt similarities with those from other regions. Phylogenetic analysis based on ORF2 region showed that the local HAstV-1 sequences were most closely related to Japanese strains, and had distinct topology with phylogenies based on ORF1a and ORF1b regions. Conclusion:The complete genome sequences of 11 HAstV strains are obtained, and the VA2 complete genome is found.