Objective:To characterize the complete genome sequence and elucidate the structural features of norovirus (NoV) isolate SD20200267.Methods:The viral nucleic acid was extracted from patient samples, followed by amplification and sequencing for genotyping based on the nucleotide sequences. The metagenomic sequencing technology was utilized for whole genome sequencing, and subsequent analysis was performed on the acquired nucleotide sequences.Results:The complete genome sequence of the SD20200267 strain, spanning a total length of 7 465 nucleotides, was successfully obtained. The SD20200267 strain belongs to the GⅡ.12 and GⅡ.P16 genotypes in the VP1 and RdRp regions, respectively. The nucleotide sequence identity of SD20200267 strain with other GⅡ.12[P16] strains ranged from 96.0% to 97.3%, exhibiting 15 amino acid variations. The strain displayed evidence of recombination, with the recombination site located in the overlapping region of ORF1 and ORF2.Conclusions:SD20200267 is classified as a GⅡ.12[P16] strain, and recombination was observed in the overlapping region of ORF1 and ORF2.

Objective:To study the infection of human rhinovirus, respiratory syncytial virus and human adenovirus in Jinan from April 2018 to March 2019, and analyze epidemiological characteristics of human adenovirus.Methods:All of 1969 nasopharyngeal swabs were collected from hospitalized patients with acute respiratory tract infections in The Fourth People’s Hospital of Jinan, Qilu Children′s Hospital of Shandong University, Peoples Hospital of Zhangqiu District from April 2018 to March 2019, fluorescence quantitative PCR was used to detect the positive rate of human rhinovirus, respiratory syncytial virus and human adenovirus. Seven adenovirus positive samples were isolated and examined by sequencing, then we determined adenovirus type, constructed gene phylogenetic trees for analysis.Results:Of the 1969 samples, 242 were positive, the total positive rate was 12.30% (242/1969), the positive rate was 3.00% (59/1969) for rhinovirus, 6.30% (124/1969) for respiratory syncytial virus (RSV), and 3.86% (76/1969) for adenovirus. There was no significant difference in the detective rate of rhinovirus in different age groups (Fisher′s exact test value =8.376, P=0.720), the positive rates of RSV and adenovirus in different age groups was statistically significant ( χ2=19.28, 12.16; P=0.001, 0.016). There was a statistically significant difference in the positive rate of adenovirus between different sexes ( χ2=14.33, P<0.001), and there was no statistically significant difference in the positive rate of rhinovirus and RSV between males and females ( χ2=0.30, 2.90, P=0.862, 0.089). Comparing the positive rates of viral nucleic acid in different months, we found that the positive rate of rhinovirus, RSV and adenovirus separately reached the highest in October, December and November (8.61%, 26.50% and 8.84%). We constructed a gene phylogenetic tree after seven positive samples of adenoviruses were sequenced, by the molecular typing method we detected that seven adenovirus-positive samples were all HAdV-2 type. Conclusions:By comparing the epidemiological trends of human rhinovirus, RSV and human adenovirus in Jinan from April 2018 to March 2019 in different ages, genders, and months, providing reference basis for the early prevention and clinical treatment of acute respiratory tract infection.

Objective To study bacteria-blocking effect of surgical gowns with new material.Methods Semi-quantitative and qualitative testing methods were used to detect bacteria-blocking rates of key sites of surgical gowns(chest and forearm), the detected samples included sample A (composite material, unused), sample B (composite material, after washing 100 times), and sample C (monolayer material, unused).Results In semi-quantitative testing,the average bacteria-blocking rates of three samples were 75.47%, 70.78%, and 73.73% respectively.In qualitative testing,three samples could effectively block the penetration of Staphylococcus aureus under wet condition and Bacillus subtilis var.niger spores under dry condition.Conclusion In semi-quantitative testing, all three kinds of samples had bacteria-blocking effect, and the average bacteria-blocking rate was> 70%;in qualitative testing, three samples all meet requirements of bacteria-blocking effect under wet and dry condition.

Objective PU.1 plays a key role in innate immune function in the alveolar macrophage.This study was to con-struct and identify recombinant adenovirus vectors carrying the human transcription factor PU.1 gene. Methods The recombinant shut-tle plasmid was obtained from the PU.1 gene ( SPI1) and eukaryotic expression vector that digested by restriction enzymes and connected by T4 DNA ligase.The target fragment SPI1-IRES-EGFP was amplified by PCR.The product was cloned into the intermediate pDONR221 and then recombined with the adenovirus backbone plasmid pAd/CMV/V5-DEST to form a recombinant adenovirus vector. The recombinant adenovirus vector was linearized by PacI and then transfected into human embryonic kidney (HEK293) cells to obtain the recombinant adenovirus pAD-SPI1-IRES-EGFP, which was then propagated in HEK293 cells, filtered and purified to obtain high-con-centration adenoviruses.The adenovirus titer was determined by TCID 50 assay.The PU.1 gene expression in the HEK293 cells was con-firmed by fluorescence microscopy and real-time qPCR. Results PCR amplification, restriction digestion and sequencing analysis showed the recombinant adenovirus carried the correct PU.1 gene.The final virus titer, calculated by TCID 50, was 8 ×1011 IU/mL. Green fluorescence was observed under the fluorescence microscope. Real-time qPCR confirmed that the expression of PU.1 mRNA was increased by 2189.93 folds. Conclusion The recombinant adenovirus vector carrying the PU.1 gene was constructed and obtained successfully, which could contribute to further studies of the influence of PU.1 overex-pression on the innate defense against Aspergillus fumigatus.

Objective To investigate the effect of acupuncture plus astragalus polysaccharide on the expression of Bcl-2 protein in pancreatic islet b cells in db/db mice. Method C57BL/Ksj-db/db mice as an animal model of spontaneous type 2 diabetes were selected for this experiment. Five-week-old db/db mice were randomized into model, acupuncture, medication and acupuncture+medication groups. Meanwhile, db/m mice were selected as a normal group. The acupuncture group received acupuncture at points Housanli (equivalent to Zusanli, ST36), Neiting(ST44) and Yishu(Extra) and the medication group, an oral gavage of astragalus polysaccharide (1400 mg/kg). Both groups were treated once daily, for 12 consecutive weeks. After the end of experiment, blood glucose, insulin and resistin were measured, and the expression of Bcl-2 protein in islet b cells was determined by immunohistochemical method. Result Blood glucose, insulin and resistin levels were significantly lower in the acupuncture+medication, acupuncture and medication groups than in the model group. They were significantly lower in the acupuncture+medication group than in the acupuncture and medication groups and significantly lower in the acupuncture group than in the medication group. The expression of Bcl-2 protein in islet b cells was higher in the medication, acupuncture and acupuncture+medication groups than in the model group; there was a statistically significant difference (P<0.01). The expression of Bcl-2 protein was higher in the acupuncture+medication groups than in the medication group (P<0.01) and it was basically the same in the medication group as in the acupuncture group; there was no statistically significant difference (P>0.05). Conclusion Acupuncture plus astragalus polysaccharide can significantly reduce blood glucose and serum insulin and resistin levels and increase the expression of Bcl-2 protein in islet b cells to effectively inhibit apoptosis in islet b cells in db/db mice. Its effect is better than that of acupuncture alone or medication.

The external and internal characteristics of domestic master and doctorial papers on rheumatoid arthritis were analyzed by visualization analysis, which revealed the publication of and hotspots and frontiers in domestic master and doctorial papers on rheumatoid arthritis, and can thus provide reference for the development of medical information resources and recent advances in rheumatoid arthritis for the broad masses of scientific workers.

Objective The study aimed to observe the clinical effects and the adverse reactions of recombinant mutant human tumor necrosis factor-alpha ( rhu-TNF) on the treatment of malignant pleural effusion ( MPE) induced by lung adenocarcinoma . Methods 70 patients with MPE caused by lung adenocarcinoma hospitalized in our department were chosen as the research objects . After conventional drainage of pleural effusion , the patients were divided into two groups according to the weight , respectively receiving intra pleural injection of 2 000 000 units and 3 000 000 units of rhu-TNF, followed by the observation of clinical effects and adverse reac-tions.A further retrospective analysis were made on the effects of dexamethasone injected before operation on clinical effects and ad -verse events. Results The effective rates of 2 000 000 unit group and 3 000 000 unit group were respectively 75.68%and 87.88%(P=0.23), with no statistical difference.The adverse reactions in the group of patients being injected dexamethasone before operation significantly reduced(P=0.021) and the use of dexamethasone had no influence on the efficacy of rhu-TNF ( P=0 .486 ) . Conclusion Rhu-TNF is a safe drug with high efficiency in the treatment of MPE induced by lung adenocarcinoma , and the efficacy and safety of re-peated application in clinic still need more supporting data .

Objective Vaccination is a most effective method for the prevention of severe diseases caused by pandemic influenza and microRNA ( miRNA) mediated gene silencing has offered a novel approach to the construction of new vaccines.Our study aimed to construct a recombinant influenza A ( H1 N1 ) virus with the PB1 gene that carries the target fragment of miRNA Let-7b. Methods After comparing the sequence of the A/Nanjing/108/2009 H1N1 viral fragments with that of Let-7b, we selected PB1 as the optimal gene sequence, inserted the Let-7b binding target gene into PB1, ligated the modified fragments with pDP 2000, and named the recombinant plasmids pDP-mu-PB1 and pDP-sclb-PB1, respectively.We co-transfected the MDCK and 293T cells with the recombinant and other seven plasmids and injected the supernatant into the allantoic cavity of the chickenembryo for virus propagation, followed by detection of the virus by hemagglutination ( HA) assay and measurement of the viral titer by TCID50 .We amplified the viral cRNA by RT-PCR and identified the viruses by agarose gel electrophoresis and nucleotide sequence analysis. Results PB1 was the optimal sequence ( 83 bp -107bp) for the attenuation of viruses.The HA-titers of miRT-H1N1 and scbl-H1N1 were 1∶32 and 1∶64, and their viral loads were 4.68 ×105 and 7.94 ×104 TCID50/mL, respectively.Nucleotide sequence analysis showed the expected fragment in the rescued virus. Conclusion A recombinant strain vaccine was successfully constructed, which has laid the foundation for fur-ther assessment of virulence.

Objective Dendritic cell-associatedC-type lectin-1 ( Dectin-1) is one of the most important receptors in antifungal innate immune response.This study was to construct a recombinant adenovirus vector expressing themurine Dectin-1gene and acquire a high-concentration adenovirus by amplification and purification. Methods The PCR amplification product CLEC7A-pIRES2-EGFP was cloned into the intermediate vector pDONR221, and then recom-bined with the backbone vector pAD/CMV/V5-DEST to produce a re-combinant plasmid pAD-CLEC7A-pIRE2S -EGFP.The recombinant plasmid was linearized with Pac I and transfected into human embryon-ic kidney ( HEK293) cells to produce recombinant adenovirus pAD-CLEC7Ap-IRES 2-EGFP. The adenovirus was propagated in the HEK293 cells and purified by filtering through the cellulose acetate membrane and concentrating column.Fluorescence microscopy and re-al-time PCR were used to determine the expression of the Dectin-1 gene. Results PCR identification, enzyme digestion, and sequen-cing results manifested theDectin-1 gene in the vector, with the final adenovirus titer of 5×1011 IU/mL.Fluorescence microscopy revealed green fluorescence and real-time PCR assay confirmed that the expression of Dectin-1 was improved by 8677.25 times. Conclusion A relatively high-titer adenovirus expressing Dectin-1 was acquired,which may help to further study the high expression of Dectin-1 in anti-fungal innate immunity in vitro and in vivo.

Objective To compare the results of susceptibility testing by European Committee on Antimicrobial Susceptibility Testing (EUCAST)and Clinical and Laboratory Standards Institute (CLSI)broth microdilution methods against Aspergillus isolates.Methods The susceptibilities of 116 Aspergillus isolates were determined for amphotericin B, voriconazole, itraconazole,caspofungin and micafungin according to EUCAST (E.DEF 9.1 )and CLSI (M38-A2)methods.The essential agreement (EA),categorical agreement (CA),very major errors (VME)and major errors (ME)of the two methods were compared.Results The EA was 96.3%-100% between the two methods.The CA ,ME,and VME were 98.8%,0-1.2% and 0 respectively for the susceptibility of Aspergillus fumigatus to voriconazole.The CA,ME and VME was 1 00%,0 and 0 respectively for the susceptibility of Aspergillus fumigatus and Aspergillus niger to amphotericin B,or the susceptibility of Aspergillus fumigatus and Aspergillus flavus to itraconazole.Conclusions The results of susceptibility testing by EUCAST and CLSI broth microdilution methods are well consistent against Aspergillus isolates.