Objective To develop a double antibody sandwich ELISA method with neutralizing antibody for the determination of content of functional antigen diphtheria toxoid(DTd), and to verify and preliminarily apply it to the quality analysis in the DTd production.Methods The concentration of anti-diphtheria toxin(DTx) coating antibody and enzyme-labeled antibody in double antibody sandwich ELISA was determined by square titration, and a linear standard curve was established.The method was verified according to the requirements of Chinese Pharmacopoeia(Volume Ⅳ, 2020 edition), and was preliminarily applied to the detection of DTd stock solution.Results The optimal concentration of coating antibody was 5 μg/mL,the working dilution of enzyme-labeled antibody was 1∶5 000 and the best linearity of dose-response curve was in a range of 0. 000 781-0. 012 5 Lf/mL(r > 0. 99). This method showed no cross reaction with pertussis toxoid(PTd), filamentous haemagglutinin(FHA), pertactin(PRN), tetanus toxoid(TTd) and Sabin-strain inactivated poliomyelitis vaccine(sIPV) Ⅰ,Ⅱ,Ⅲ. For the DTd standard in the linear range, the coefficients of variation(CVs) of precision verification were 4. 23%, 2. 98%,1. 81%, 6. 61% and 1. 82%, and the recoveries of accuracy verification were 90. 67%, 105. 39%, 102. 11%, 97. 76% and 81. 31%, respectively. The Pearson r was 0. 638 0(P < 0. 05) for the determination of toxoid content in 12 batches of DTd stock solution by this method and flocculation test. For the concentration of diphtheria toxoid less than 1 800 Lf/mL, the Pearson r was 0. 899 2(P < 0. 001).Conclusion A double antibody sandwich ELISA method for quantification of DTd antigen based on neutralizing antibody was successfully developed with good specificity, accuracy and precision, which provides an effective verification means for the production of DTd stock solution.

Objective To investigate the clinical effect of ulinastatin treating systemic inflammatory response in the patients with serious blood infection .Methods The patients with serious blood infection in our hospital were selected and divided into the control group and observation group .The control group was treated with the conventional treatment ,while on this basis the observation group was added with ulinastatin .The body temperature ,heart rate ,respiratory rate ,WBC ,Scr ,CRP ,PCT ,TNF-α,APACHE Ⅱscore and mortality rate were compared between the two groups .Results The body temperature ,heart rate ,respiratory rate and A-PACHE Ⅱ score in the observation group were lower than those in the control group ,moreover ,the levels of WBC ,CRP ,PCT , TNF-αand Scr ,and mortality rate in the observation group were lower than those in the control group ,the differences were statisti-cally significant (P<0 .05) .Conclusion Uinastatin has good effect for treating systemic inflammatory response in the patients with serious blood infection ,which can decrease the mortality and is worth clinical promotion .

Objective To analyse the prognosis of 117 newly diagnosed nasopharyngeal carcinoma (NPC) patients underwent intensity modulated radiotherapy (IMRT). Methods From Jan to Nov 2005, 117 NPC patients who were treated by IMRT were enrolled. There were 81 males and 36 females with a median age of 42 years (range 18-76 years). According to Chinese Fuzhou Staging system(1992), 11 cases were Stage I , 15 Stage Ⅱ, 54 Stage Ⅲ and 37 Stage ⅣA. IMRT was carried out with Peacock plan. The prescription dose to the gross target volume(GTVnx) of nasopharyngeal tumor was 68 Gy, that of positive neck lymph nodes (GTVnd) was 60-66 Gy, clinical target volume 1 (CTV1) was 60 Gy, and CTV2 was 54 Gy. Results After a median follow-up time of 48 months (range 10.5-59.5 months), the 3-and 5-year overall survival (OS) rates were 95.7 % and 89.7 %, the disease-free survival (DFS) rates were 91.5 % and 87.2%, and the local-regional control rates were 94.0 % and 91.5 %. Univariate analysis showed the KPS, stage, Fuzhou clinical stage, status of blood platelet before treatment and uric acid after treatment were correlated with OS rate. T stage was the only independent factor of prognosis in the COX stepwise regression model. Conclusion Radical IMRT significantly prolongs the survival of NPC patients. T stage is the only independent prognostic factor for NPC patients.

Gastric cancer, a common malignant tumor in digestive system with high morbidity as well as mortality rate, is insidious at the onset and lack of effective treatments so far. A growing number of studies have shown that exosome-derived miRNAs play an important role in the occurrence and development of gastric cancer. Autocrine exosome miRNAs from gastric cancer cells regulated tumor growth, recurrence, metastasis and drug resistance, etc. Moreover, exosomal miRNAs in the tumor microenvironment can be delivered into cancer cells to facilitate intercellular communication, thus affecting the progress of gastric cancer. Due to exosomes, which were released into circulation from tumor cells, contain abundant, specific and stable miRNAs, exosome-derived miRNAs have a great potential to be used as novel diagnosis biomarkers and treatment targets of gastric cancer.(Chin J Lab Med, 2018, 41: 499-502)

Objective: To understand fruit and vegetable consumption of children and associated factors during COVID-2019 control period, and to provide basis for growth and development of children. Methods: In February 2020, 334 parents of children aged 2 to 15 years old from 22 provinces, cities and autonomous regions, including Hubei, Beijing, Shandong, Gansu were surveyed through the "Questionnaire Star" network questionnaire for the students’ fruit and vegatable consumption during the period of COVID-19, and Logistic regression analysis was conducted with multivariate analysis. Results: Among the children, 26.4% of their vegetable consumption and 45.2% of their fruit consumption had changed. The multivariate analysis showed that inconvenient purchase (OR=6.52) were related to the decrease of children’s vegetable consumption. The 2-5 age group(OR=0.28), whether parents actively gained relevant knowledge of diet (OR=0.33) and mild epidemic risk (OR=4.35) were related to the increase of children’s vegetable consumption(P<0.05). Average monthly income of family(OR=6.27-8.39), inconvenient purchase(OR=2.93), and epidemic risk (OR=0.21-0.26) were associated with reduced fruit consumption among children(P<0.05). While gender(OR=2.20), average monthly income of family below 2 000 yuan (OR=6.94) and mild epidemic risk (OR=0.35) were associated with increased fruit consumption among children(P<0.05). Conclusion Children’s fruit and vegetable consumption were greatly affected by the inconvenience of food purchase during the period of COVID-19. Low-income families were less resilient, and their children’s food consumption was more likely to be affected. Parents’ high level of nutritional knowledge could help weaken the adverse impact of the epidemic on children’s diet.

Malignant tumors of the digestive system , with high morbidity and mortality rate , are insidious at the onset and lack of effective treatments so far .Interleukin-22 (IL-22) is one of the members of the IL-10 cytokine family discovered in recent years and was originally called IL-10-associated T cell differentiation inducing factor (IL-TIF).IL-22 expression is elevated in various digestive system malignant tumors, and increased IL-22 expression is associated with tumor progression and poor prognosis .Studies on the molecular mechanism revealed that IL-22 initiates a series of downstream signaling pathways such as JAK/STAT and MAPK, by acting on the IL-22 receptor, inducing tumorigenesis.IL-22 is expected to be a novel diagnostic biomarker and therapeutic target of digestive system malignant tumor .

Objective·To further clarify the mechanism of podocyte damage by studying the expression of cartilage oligomeric matrix protein(COMP)in glomerular podocytes and its relationship with podocyte autophagy under high glucose environment.Methods·The gene expression dataset GSE104948 was downloaded from the GENE EXPRESSION OMNIBUS(GEO)database,and differentially expressed genes(DEGs)were obtained via GEO2R.The molecular functions and signaling pathways related to differential genes were summarized.The most correlated key genes(hub genes)were acquired by Weighted Gene Co-Expression Network Analysis(WGCNA)and the protein-protein interaction network(PPI)of DEGs was constructed with STRING database.The enrichment analysis was performed again.Conditionally immortalized mouse podocyte cells were cultured in vitro.After being fully differentiated,they were stimulated with high glucose,and the expressions of COMP,mammalian target of rapamycin(mTOR),microtubule-associated protein 1 light chain3(LC3)and other proteins in podocytes were detected by Western blotting.The shRNA constructed by lentiviral vector was further used to infect podocytes to inhibit the expression of COMP,and the stable cell strains were screened by puromycin.The expression of COMP,mTOR,and LC3 of stable strains were detected by Western blotting,in order to observe the effect of COMP on autophagy.Results·A total of 362 DEGs were filtered for subsequent analysis.Among these DEGs,284 genes were up-regulated and 78 genes were down-regulated.The results of Gene Onotology(GO)term analysis showed that DEGs in diabetic nephropathy(DN)were mainly enriched in cell surface receptor signaling pathway,receptor binding,etc.The main enriched Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways included phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/AKT)signaling pathway,extracellular matrix(ECM)-receptor interaction,etc.Sixty-four hub genes were refined through the intersection of WGCNA and PPI hub genes,and the hub genes with significantly increased or decreased expression were sifted.The hub genes were annotated with KEGG again,and it was found that most of the hub genes were enriched in"ECM-receptor interaction"and"PI3K/AKT signaling pathway".The PI3K/AKT/mTOR signaling pathway is a classic autophagy pathway,and COMP was absolutely overexpressed(logFC>2)in the 64 hub genes,suggesting that it may affect autophagy through this pathway.Western blotting showed that compared with the mannitol control group and the low glucose group,the expression of COMP in podocytes was significantly increased under high glucose stimulation.Compared with the control group,the expression of LC3-Ⅱ in the high glucose group was significantly decreased,indicating that the autophagy initiation of podocytes was inhibited under the high glucose environment.Compared with the negative control,the expression of LC3-Ⅱ in renal podocytes of mice with knockdown of COMP was significantly increased,and the mTOR decreased with the decrease of the expression of COMP,indicating that inhibiting COMP contributed to the recovery of autophagy in podocytes.Conclusion·COMP is highly expressed in DN patients and highly enriched in ECM receptor and PI3K/AKT signaling pathway.Autophagy in mouse renal podocytes is inhibited under high glucose conditions,and the high expression of COMP induced by high glucose may be a key factor in autophagy inhibition.Inhibiting COMP helps to restore autophagy in mouse renal podocytes.