Objective:To prepare IL-12 modified with polyethylene glycol derivatives and determine its modification sites,and to evaluate it in vitro in terms of stability,hemogram recovery,anti-tumor effects and other aspects.Methods:IL-12 was modified with polyethylene glycol propionaldehyde and maleimide imide derivatives,and modified sites of two different derivatives were deter-mined by ultra performance liquid chromatography(UPLC).Recombinant human IL-12 and PEG-IL-12 were used to stimulate NK92 cells,activity and cytotoxicity of IFN-γ were evaluated by kits.CD34+was stimulated to evaluate its blood picture recovery potential.NK cell killing was stimulated to evaluate its effectiveness in improving immunity and anti-tumor effects.Results:Compared with recombinant human IL-12,PEG-IL-12 had higher stability,blood picture recovery potential and anti-tumor effects.Conclusion:PEG-IL-12 can effectively overcome many drawbacks of recombinant human IL-12,greatly improving possibility of its widespread applica-tion in clinical trials.

[Objective]To introduce and summarize Professor LI Xinmin's experience in treating pediatric epilepsy from the perspective of pivot.[Methods]By following the clinical work,it collected,organized and analyzed initial and follow-up medical records of pediatric epilepsy patients from the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine and consulted ancient Chinese medicine books and modern clinical research on pediatric epilepsy,and analyzed Professor LI's experience in treating pediatric epilepsy from several aspects.Additionally,it attached a medical case for verification.[Results]Professor LI identifies phlegm turbidity and retention as the basic initiating factor of the disease,and phlegm Qi disorder as the core pathogenesis.Combining with the characteristics of children,he utilizes the characteristics of Chinese medicine,using the pivot as the core to treat phlegm and promote Qi flow,supplemented by reconciling the Shaoyang pivot,restoring the circulation of spleen and stomach,and using a dual approach to regulate the overall Qi and coordinate the functions of organs,thereby improving the condition of children with epilepsy and reducing the frequency of seizures.Professor LI also pays attention to individualized treatment,examines the evidence to determine the cause,and adds or subtracts medication according to different types of seizures.At the same time,emphasis is placed on dynamic syndrome differentiation,and medication is prescribed according to the changes in the patient's condition,achieving good therapeutic effects in the treatment and prevention of epilepsy.The attached case was diagnosed as disadvantageous of Shaoyang pivot syndrome at first visit,the treatment was to dredge and benefit Shaoyang,calm the liver and extinguish wind.Chaihu Longgu Muli Decoction and Ditan Decoction were added and subtracted,and then Xieqing Pill was given to clear the liver meridian and remove the heat.[Conclusion]Professor LI's clinical efficacy in treating pediatric epilepsy from the perspective of pivot theory is affirmed,which enriches the diagnosis and treatment system of pediatric epilepsy and is worthy of learning and reference.

Objective:To investigate the whole genome of SARS-CoV-2 causing COVID-19 in Rongcheng city of Shandong Province in May 2022 and to further analyze the nucleotide and amino acid variations for source tracing.Methods:High-throughput sequencing was used to sequence the SARS-CoV-2 genome in 15 nasopharyngeal swab samples from COVID-19 cluster infections and three environmental samples related to an aquatic product import company. Whole-genome sequence splicing, variant site analysis and sequence typing were performed on the raw sequencing data using virus sequence and variant analysis software. A phylogenetic tree was constructed by evolutionary analysis software. Epidemiological investigation was used to trace the potential source of infection.Results:Thirteen whole genome sequences of SARS-CoV-2 with the length ranging from 29 653 bp to 29 780 bp were successfully obtained from the nasopharyngeal swab samples. The average sequencing depth was 1 756-6 565 X and the genome coverage was 99.20%-99.63%. The results of Pangolin typing showed that the 13 genomes belonged to the VOC/Gamma (P.1.15) evolutionary branch. Compared with the Wuhan reference strain (NC_045512.2), the 13 genome sequences had 40-41 nucleotide mutation sites. There were 23-24 amino acid variation sites in seven protein domains (ORF1a, ORF1b, S, ORF3a, ORF8, ORF9b and N proteins). Evolutionary analysis showed that the viral sequence was grouped to the same subclade as the reference strain from Argentina (EPI_ISL_4082233).Conclusions:In this study, the whole genome sequences of 13 Gamma variant strains were obtained from COVID-19 cluster infections associated with imported cold-chain aquatic products in Rongcheng city, and the imported seafood from South America in 2021 was found to be the source of the virus in a timely manner. This study provided reference for the SARS-CoV-2 variant analysis and case tracing and also suggested that the survival and transmission ability of SARS-CoV-2 on the surface of cold-chain products should not be underestimated and needed further investigation.

Objective:To identify the genome-wide characteristics and variations of the novel coronavirus 2019(2019-nCoV) by combining high-throughput sequencing and bioinformatics techniques, to type and trace the virus.Methods:Pharyngeal swab sample of one 2019-nCoV infection case imported from Weihai city was selected for 2019-nCoV whole-genome sequence capture and NGS high-throughput sequencing. The obtained sequencing data were sequenced and analyzed using virus mutation analysis software, we classify viruses and build phylogenetic trees to track their potential sources.Results:The complete genome sequence of 2019-nCoV was obtained, with a total length of 29 808 bp and an average sequencing depth of 5 013.11×. A total of 51 nucleotide mutations were found in 8 coding regions (ORF1ab, S, ORF3a, E, M, ORF6, ORF7a and N). The result of virus typing showed that the virus belonged to Omicron BA.2. Evolutionary analysis showed that the virus sequence was in the same clade as the reference strains from Japan.Conclusions:The sequencing method and analysis result established in this study can be used for mutation analysis of 2019-nCoV and case tracing, which is of great significance for the prevention and control of COVID-19.

Objective To analyze the malaria epidemiological characteristics in Tengchong City from 2010 to 2015,so as to provide the evidence for adjusting and formulating measures in the elimination stage. Methods The malaria data were collected and analyzed by using Microsoft Excel 2010 in Tengchong City from 2010 to 2015. Results There were 1 408 malaria cases re?ported in Tengchong City from 2010 to 2015,including 1 091 cases of Plasmodium vivax infection,256 cases of P. falciparum infection,5 cases of P. malariae infection,1 case of P. ovale infection,1 case of mixed infections,and 54 unclassified cases. Totally 1 390 imported cases were recorded and 98.06%of them(1 363/1 390)were imported from Myanmar. Most of the pa?tients(n=908)were aged from 21 to 40 years,and the male to female ratio was 11.03∶1. The highest?risk populations were farmers and migrant workers. The most cases were observed in April and June,and at that time,most of the floating workers re?turned. Conclusion Imported malaria is severe in Tengchong City,and there is a great challenge to malaria elimination.

Objective To describe the level of self-perceived burden (SPB) and analyze its influencing factors among the patients with advanced lung cancer, in order to provide a theoretical basis for individualized nursing care. Methods A total of 102 hospitalized patients with lung cancer from 3 hospitals in Shandong province were investigated by the Self-Perceived Burden Scale for Cancer Patient (SPBS-CP). Results The total score of the SPBS-CP was (60.31±17.06) points, which indicated at a moderate level. Those patients rated the highest score on the sub-scale of economic/family burden (22.04±6.72) points, the lowest score on the sub scale of care burden (10.28±3.62) points. Single factor analysis and regression results showed that different work conditions, different stages were related with SPB. Conclusions Patients with moderate and advanced lung cancer had a moderate or higher level of SPB. Staff nurses should focus on the psychological status of the patients and take positive interventions to release the negative emotional experiences.

OBJECTIVETo investigate the effect and mechanism of high dose Vitamin B3 on granulopoiesis in normal rat.

METHODSTwenty one healthy SD rats were randomly divided into three groups: the Vitamin B3 group (Vit B3 500 mg·kg⁻¹·d⁻¹, × 7 d), the rhG-CSF group (rhG-CSF 25 μg·kg⁻¹·d⁻¹, × 7 d) and the normal saline group (2 ml/d, × 7 d). The peripheral blood cell counts were analyzed by automatic blood cell counter before (day 0) treatment, the third day (day 3) and the seventh day (day 7) after administration of drugs, respectively. The concentration of serum nicotinamide adenine dinucleotide (NAD⁺) level was measured by enzymatic cycling assay before and after drugs treatment. The expressions of G-CSF, G-CSFR, SIRT1, C/EBPα, C/EBPβ, C/EBPε and NAMPT mRNA were detected by reverse transcription real-time fluorescent quantitative PCR.

RESULTSThe neutrophil counts increased significantly after 7 days of Vitamin B3 and rhG-CSF treatment compared with that of control group [(1.64 ± 0.19) × 10⁹/L, (1.88 ± 0.37)× 10⁹/L vs (0.86 ± 0.18) × 10⁹/L, P<0.01]; the level of serum NAD⁺ increased significantly [(0.96 ± 0.08) nmol/L, (0.65 ± 0.12) nmol/L vs (0.36 ± 0.15) nmol/L, P<0.01]; the expression of G-CSF, G-CSFR, SIRT1, C/EBPα, C/EBPε and NAMPT mRNA in bone marrow mononuclear cells were increased significantly compared with that of control group (P<0.01).

CONCLUSIONHigh dose of Vitamin B3 may play an important role in increasing absolute neutrophil count in healthy rat under steady state, and the mechanism may be dependent on NAMPT-NAD⁺-SIRT1 signaling pathways.

Animals ; Bone Marrow Cells ; Granulocyte Colony-Stimulating Factor ; Leukocyte Count ; Neutrophils ; drug effects ; Niacinamide ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins

Objective To investigate the clinical features and pathogenesis of severe congenital neutropenia (SCN) by detecting the gene mutation of a SCN patient suspected by clinical diagnosis. Methods The intravenous anticoagulant and clin-ical data and laboratory results of this child were collected;the phagocyte and oxidation function of neutrophils were evaluated by flow cytometry;ELANE, HAX1, WAS, GFI1, CSF3R and CXCR4 genes were screened by PCR amplification and sequencing. Results The neutrophil function of this patient was normal; sequencing results revealed no mutation occurred in ELANE, HAX1, WAS, GFI1, CSF3R and CXCR4;and granulocyte colony-stimulating factor (G-CSF) can obviously enhance the level of neutrophils. Conclusion SCN is a kind of genetic heterogeneity syndrome associated with multiple gene mutations, gene diag-nosis will contribute to understanding of the pathogenesis of the disease and provide theoretical basis for treatment. Though more and more pathogenic genes were found to be connected with SCN, the cases of unknown mutation still account for a large proportion of this disease.

Objective To screen the sequence of fibroblast growth factor receptor 3 (FGFR3) genes in children with dys-chondroplasia and their family members for searching the mutations. Methods The sequence of exon 10 and exon 13 in muta-tion hot spot region of FGFR3 gene in seven families was analyzed using polymerase chain reaction (PCR) and DNA sequenc-ing technology. Results The c.1138G>A missense mutation in exon 10 was found in 4 probands who were diagnosed as achon-droplasia (ACH), while this mutation was absent in their parents. The c.1620C>A missense mutation in exon 13 was found in one girl and her mother who both were diagnosed as hypochondroplasia (HCH) with mild symptoms. Neither mutation men-tioned above was found in the other two probands. Conclusions Through detecting the mutation in exon 10, exon 13 of FGFR3 gene, most patients of ACH or HCH can be finally diagnosed. However, it is necessary to perform the mutation screening on the other zones of FGFR3 gene and on other related genes for a few cases.

Obj ective To discuss yhe influence of nursing inyervenyion on complicayions of liver yumor payienys wiyh meyal yrack marks implanyayion.Methods 160 payienys wiyh meyal yrack marks implanyayion were randomly divided inyo inyervenyion group and conyrol group.Each group conyained 80 payienys.The payienys in yhe conyrol group were given convenyional nursing of meyal yrack marks implanyayion.Healyh educayion was given in yhe inyervenyion group on yhe basis of con-venyional nursing.Incidence raye of complicayions such as yachycardia afyer operayion,hyperyen-sion,abdominal bleeding and pain,bleeding and skin pain on yhe puncyure poinys were compared beyween yhe ywo groups.Results Afyer meyal yrack marks implanyayion,160 payienys finished yheir cyberknife yreaymeny wiyhin 5~7 days.The incidence raye of complicayions,such as yachy-cardia afyer surgery,hyperyension,abdominal bleeding and pain,bleeding and skin pain of yhe puncyure poinys in yhe inyervenyion group were obviously lower yhan yhay in yhe conyrol group,yhe difference was syayisyically significany(P<0 .05 ).Conclusion Implemenyayion of healyh educayion before and afyer yhe surgery,paying ayyenyion yo yhe change of payienys′disease progression,psy-chology and behavior,performing nursing inyervenyion ay yhe righy yime and carefully deyecying yhe influence on payienys can effecyively lower yhe incidence of complicayions.