CD20 is a surface marker protein of B-cell lymphoma, and its extracellular region is the target of specific antibodies and drugs. To obtain a cheap and easily modified specific preparation targeting CD20, we optimized the gene of CD20 extracellular region according to codon degeneracy to facilitate its expression in Escherichia coli. The optimized gene was cloned into pGEX-4T-1 vector, and the recombinant vector was transformed into E. coli BL21(DE3) for expression. The purified protein was identified by SDS-PAGE and Western blotting. Systematic evolution of ligands by exponential enrichment (SELEX) was employed to screen the ssDNA aptamer that specifically binds to the fusion protein, and the affinity of the aptamer to CD20 was detected by flow cytometry. Then, the cytotoxicity test was carried out to examine the inhibitory effect of the aptamer on B lymphoma cells. In this study, we established the prokaryotic expression method of CD20 and obtained the aptamer specifically binding to the extracellular region of CD20, which laid a foundation for the development of therapeutic drugs targeting CD20.
Humans ; Escherichia coli/metabolism* ; SELEX Aptamer Technique/methods* ; Aptamers, Nucleotide/genetics* ; Antigens, CD20/metabolism* ; Ligands

ObjectiveTo investigate the prevalence and its influencing factors of hyperuricemia (HUA) among the employed individuals who undergo occupational health screening in Shanghai, so as to provide a scientific basis for the prevention and control of HUA and for the health promotion in this population. MethodsCluster sampling methods were performed to recruit all the employed individuals who underwent routine health check-ups at the Shanghai Health and Medical Center from January to December 2023. Relevant data, including physical examination results, laboratory tests, and questionnaire surveys, were collected for analyses. Based on the diagnostic criteria for HUA, participants were categorized into HUA group and non-HUA group. Logistic regression analysis was used to identify the influencing factors of HUA. ResultsThe detection rate of HUA in the employed population of Shanghai was 23.26%, with a significantly higher rate in males (31.12%) than that in females (10.43%) (χ²=381.853,P<0.001). The detection rate of HUA increased with age (χ²_trend=5.728, P_trend=0.017). Logistic regression analysis revealed that males had a higher risk for HUA the females, with overweight, obesity, meat-predominant diet, hypertension, and dyslipidemia being positively correlated with HUA risk. While, individuals aged 40‒ and 50‒ years had a lower risk for HUA compared with those aged 20‒ years. ConclusionThe prevalence of HUA among the employed individuals who undergo health check-ups in Shanghai is relatively high. Attention should be paid to maintaining a healthy lifestyle and keeping a balanced diet. Targeted health management should prioritize for males, individuals with hypertension and BMI overweight and obesity, and those with dyslipidemia to reduce the risk of HUA.

Objective:To discuss the expression and clinical significance of inositol polyphosphate-4-phosphatase type Ⅱ(INPP4B)gene in hepatocellular carcinoma(HCC)based on The Cancer Genome Atlas(TCGA)database and experimental verification with clinical samples.Methods:Based on data from 424 clinical samples in the TCGA database(including 374 HCC tissues and 50 paracarcinoma tissues),Kaplan-Meier method and Cox regression analysis were used to evaluate the relationship between INPP4B gene and the clinical characteristics and survival prognosis of the HCC patients.The correlations between INPP4B gene and the number of 24 types of immune cells,matrix,immune cell infiltration and tumor purity in tumor tissue,and the expression level of the high-frequency mutant gene tumor protein 53(TP53)in HCC were analyzed.The clinicopathological data and paraffin-embedded tissue sections of 60 HCC patients treated with surgical resection from December 2022 to December 2023 were collected.According to clinical diagnosis,they were divided into poorly differentiated group(HCC-L group),moderately differentiated group(HCC-M group)and well-differentiated group(HCC-H group),with 20 cases in each group;20 patients during the same period who underwent biopsy and were pathologically diagnosed as non-tumor were selected as normal group,and their clinicopathologic data and liver tissue paraffin sections were collected.HE staining was used to observe the pathomorphology of HCC tissue and normal liver tissue of the subjects in various groups;immunohistochemistry method was used to detect the expressions of Ki-67 and INPP4B proteins in the HCC tissue and normal liver tissue of the subjects in various groups.Results:The TCGA database analysis results showed that compared with normal tissue,the expression level of INPP4B mRNA in HCC tissue was significantly increased(P<0.01).Compared with INPP4B low expression group,the overall survival(OS)of the patients in INPP4B high expression group was significantly prolonged(P<0.05).The univariate Cox regression analysis results showed that tumor stage,pathological stage,tumor status and residual tumor had impacts on OS of the HCC patients(P<0.05).The univariate regression analysis results showed that the INPP4B prognostic risk model score ratio was HR=0.781,95％confidence interval(CI):0.552-1.105,P=0.168.The AUC value for the impact of INPP4B on OS of the HCC patients was 0.558,indicating that the INPP4B gene prognostic risk model had certain predictive value in survival prognosis.The INPP4B mRNA expression level was not correlated with TNM stage,stage,patient gender,age,race or body mass index(BMI)(P>0.05).In tumor tissue with high and low INPP4B expression,22 types of immune cells showed statistically significant differences(P<0.05);the INPP4B mRNA expression level was positively correlated with the number of 23 types of immune cells except T helper(Th)17 cells(r>0),among which all Th cells except natural killer(NK)CD56+cells were statistically significant(P<0.01);INPP4B was significantly correlated with matrix(r=0.475),immune cell infiltration(r=0.641)and tumor purity(r=0.599)in tumor tissue(P<0.01).INPP4B was correlated with TP53(r=0.287,P<0.01).The HE staining results showed that clear and complete lobular structure,neatly arranged cells and slight inflammatory cell infiltration were observed in liver tissue of the subjects in normal group;completely destroyed lobular structure,significant hepatocellular steatosis,massive inflammatory cell infiltration,and lesions such as ballooning degeneration and small cell hyperplasia in some cells were observed in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups,and the lower the HCC differentiation degree,the more severe the tissue destruction;The immunohistochemistry results showed that compared with normal group,the expression levels of Ki-67 protein in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups were significantly increased(P<0.01),and the lower the differentiation degree of the HCC patients,the higher the Ki-67 positive rate.Brownish-yellow granules evenly distributed in the cells and INPP4B protein was highly expressed in liver tissue of the subjects in normal group;compared with normal group,the expression levels of INPP4B protein in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups were significantly decreased(P<0.01),and the lower the differentiation degree of the HCC tissue,the lower the INPP4B positive rate.Conclusion:INPP4B is a protective factor for the prognosis of HCC patients;as a new tumor suppressor gene,INPP4B may become a potential target for new drug screening in HCC treatment.

Polycystic ovary syndrome （PCOS） is a common endocrine disorder among women of childbearing age. Persistent chronic inflammation can promote the progression of PCOS， thereby causing serious impacts on women’s endocrine metabolism， reproductive function， and psychological status. Nuclear factors- κB （NF- κB） signaling pathway is one of the most classic inflammatory response transduction pathways and is closely related to the pathological process of PCOS. This article summarizes the mechanism of traditional Chinese medicine intervention in PCOS based on the NF-κB signaling pathway， and finds that various monomers of traditional Chinese medicine [flavonoids （naringenin， soy isoflavone， rhamnocitrin， etc.）， alkaloids （berberine， ligustrazine） and terpenoids （artesunate， cryptotanshinone）， glycosides （salidroside， glycyrrhizic acid）] and traditional Chinese medicine compounds [formula for tonifying kidney （Bushen huayu formula， Bushen huatan formula， etc.）， formula for eliminating damp and dissolving phlegm （Cangfu daotan decoction， Xiehe yin， Erchen decoction）， formula for regulating blood circulation and removing stasis （Gexia zhuyu decoction， Huatan tongmai yin）] can alleviate inflammatory response， improve abnormal glucose and lipid metabolism， and enhance ovarian function by inhibiting NF-κB signaling pathway， thus improving PCOS.

Objective To explore the effect of improved total cavity endoscopy on lung function,postoperative feeding and complications in patients with chronic lung disease of esophageal cancer.Methods 120 esophageal cancer patients with chronic lung disease admitted to our hospital from September 2022 to June 2023 were randomly divided into two groups.The observation group consisted of 60 patients who underwent improved total cavity endoscopy assisted esophageal cancer resection,while the control group consisted of 60 patients who underwent traditional open surgery.The perioperative related indexes,lung function indexes[forced expiratory volume in the first second(FEV1),forced vital capacity(FVC),maximum ventilation volume(MVV)],inflammatory level[interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor(TNF-α)]were compared between the two groups.Results The blood loss,operation time,number of lymph node dissection and drainage time in the observation group were(329.51±78.84)ml,(175.47±10.41)min,(29.67±17.86)pieces per case and(3.14±0.98)d respectively.In the control group,the intraoperative blood loss,operation time,number of lymph node dissection and drainage time were(372.31±99.23)ml,(148.54±10.68)min,(28.36±18.15)pieces and(6.37±1.23)d,respectively,there was statistical significance between the two groups(P＜0.05).The FEV,,FVC and MVV of the observation group were(1.88±0.53)L,(2.33±0.46)L and(32.59±11.84)L,respectively.Two weeks after operation,the control group was(1.37±0.31)L,(1.75±0.38)L and(23.68±9.41)L respectively,there was statistical significance between the two groups(P＜0.05).The inflammatory factors IL-6,IL-8 and TNF-α in the observation group were(2.17±1.62)ng/ml,(2.09±1.52)ng/ml and(1.32±0.57)ng/ml,respectively.The control group were(3.06±1.52)ng/ml,(2.75±1.29)ng/ml and(1.73±0.75)ng/ml respectively,there was statistical significance between the two groups(P＜0.05).The incidence of postoperative complications in the observation group and the control group were 6.67％and 20.00％respectively,and patients in the observation group ate earlier than those in the control group(P＜0.05).Conclusion Compared with the traditional open surgery,the improved total laparoscopic surgery has the advantages of less trauma,simpler operation,less damage to lung function,significantly lower incidence of postoperative complications,and shorter postoperative eating time.

Objective:To investigate damaging effects of clomifene citrate(CC)on endometrial stromal cells(hEndoSCs),and to study effects of spermidine on autophagy and inflammatory cytokine expression in damaged endometrial stromal cells.Methods:Groups were firstly divided into control group,spermidine group,clomiphene group(CC group),CC+Spermidine group.MTT assay was used to detect cell survival rate of hEndoSCs after co-incubation with different concentrations of CC or Spermidine for 24 h.Con-tent of intracellular reactive oxygen species(ROS)and level of apoptosis in cells of the 4 groups were detected by flow cytometry tech-nique.Western blot was used to detect expressions of autophagy pathway-related proteins ULK1,p-ULK1,LC-3Ⅱ,and apoptosis-re-lated proteins Bax,Bcl-2,Cleaved-caspase 3.RT-qPCR was used to detect mRNA expressions of IL-6,IL-1β and TNF-α.Results:Compared with control group,CC group showed decreased cell survival,increased apoptosis rate,ROS content,Bax,Cleaved-cas-pase 3 expressions,decreased Bcl-2 expression,decreased levels of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ,and elevated expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA(P<0.01).There was no significant changes viability of cells in spermidine group compared with control group(P>0.05).Compared with CC group,cell survival rate in CC+spermidine group was sig-nificantly increased,apoptosis rate,ROS content,Bax and Cleaved-caspase 3 expressions were decreased,Bcl-2 expression was in-creased,expressions of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ were elevated,while expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA were decreased(P<0.01).Conclusion:CC can inhibit endometrial stromal cell proliferation,promote apoptosis,and increase the transcript levels of inflammatory factors IL-6,IL-1β and TNF-α.Spermidine can reduce intracellular ROS in clomiphene-injured endometrial stromal cells by activating cellular autophagy,increase cell survival,and inhibit the expressions of inflammatory factors IL-6,IL-1β and TNF-α.

ObjectiveTo determine the trend in diabetes-related death and probability of premature mortality among residents in Yangpu District of Shanghai and provide evidence for the formulation of relevant intervention strategies. MethodsMortality and demographic data were collected among residents in Yangpu District of Shanghai from 2002 to 2020. Statistical analysis was conducted using chi-square test with SPSS 21.0 and Excel 2010. Joinpoint regression was used to determine annual percent change （APC）. The mortality was standardized by utilizing the world standard population in 2000. ResultsA total of 25 091 cases of diabetes-related deaths were reported in Yangpu District， Shanghai from 2002 to 2020. The average annual crude mortality of diabetes-related diseases was 122.10/10⁵， which was 116.13/10⁵ in males and 128.23/10⁵ in females. The difference between males and females in crude mortality was statistically significant （P<0.05）. Moreover， primary causes of diabetes-related deaths were diabetes， cardiovascular and cerebrovascular diseases， and tumors. Among diabetic deaths， peripheral circulatory complications accounted for 50.79%， followed by renal complications （16.05%）. The crude mortality in males， females， and total of diabetes-related diseases showed an upward trend， while the standardized mortality remained stable with an upward trend in male and a downward trend in female. Furthermore， the crude mortality in males， females， and total of diabetes complicated with cardiovascular and cerebrovascular diseases showed an increasing trend. In contrast， both the standardized mortality in males and in total showed an increasing trend， while that in females remained stable. The overall crude mortality of diabetes was on the rise， which was increasing in males and stable in females. The overall standard mortality of diabetes was on the decline， which was increasing in males while declining in females. In addition， the probability of premature mortality caused by diabetes-related diseases， cardiovascular and cerebrovascular diseases complicated with diabetes， and diabetes decreased from 2002 to 2020 with no statistical significance. Males showed an upward trend while females showed a downward trend. ConclusionThe mortality of diabetes-related diseases， cardiovascular and cerebrovascular diseases complicated with diabetes， and diabetes among residents in Yangpu District of Shanghai is on the rise. Similarly， standardized mortality and probability of premature mortality in males for all three diseases are also on the rise. It warrants more attention to the health of male diabetes patients and targeted measures to reduce the disease burden.

Peripheral bacterial infections without impaired blood-brain barrier integrity have been attributed to the pathogenesis of Parkinson's disease (PD). Peripheral infection promotes innate immune training in microglia and exacerbates neuroinflammation. However, how changes in the peripheral environment mediate microglial training and exacerbation of infection-related PD is unknown. In this study, we demonstrate that GSDMD activation was enhanced in the spleen but not in the CNS of mice primed with low-dose LPS. GSDMD in peripheral myeloid cells promoted microglial immune training, thus exacerbating neuroinflammation and neurodegeneration during PD in an IL-1R-dependent manner. Furthermore, pharmacological inhibition of GSDMD alleviated the symptoms of PD in experimental PD models. Collectively, these findings demonstrate that GSDMD-induced pyroptosis in myeloid cells initiates neuroinflammation by regulating microglial training during infection-related PD. Based on these findings, GSDMD may serve as a therapeutic target for patients with PD.

Objective To investigate the relationship between sleep duration and obesity, and the risk of common chronic diseases in the occupational population in Shanghai City. Methods A total of 18 775 occupational individuals were selected as the study subjects using convenience sampling method in Shanghai City. Data on personal lifestyle behaviors and medical examination results were collected. The relationship between sleep duration and different types of obesity with dyslipidemia, hyperuricemia, hypertension, and hyperglycemia was analyzed. Results The incidence of dyslipidemia, hyperuricemia, hypertension, and hyperglycemia among the study subjects was 24.9%, 16.2%, 11.5%, and 7.3%, respectively. The incidence of these four chronic diseases were higher in individuals with central obesity and suboptimal sleep compared to the control group (all P<0.01). Multivariate logistic regression analysis showed that suboptimal sleep combined with general obesity/overweight increased the risk of dyslipidemia, hyperuricemia, hypertension, and hyperglycemia in the study subjects ［odds ratio (OR) were 2.40, 3.47, 3.30, and 2.79, respectively; all P<0.01］, after adjusting for age, gender, education level, marital status, occupation type, labor intensity, smoking, and drinking. Suboptimal sleep combined with central obesity also potentially increased the risk of these four chronic diseases (OR were 2.25, 3.09, 3.09, and 2.98, respectively; all P<0.01). Conclusion The incidence of common chronic diseases is relatively high in the occupational population in Shanghai City. Suboptimal sleep combined with different types of obesity increases the risk of common chronic diseases.

Pediatric idiopathic nephrotic syndrome (INS) is characterized by massive albuminuria, hypoproteinemia, edema and hyperlipidemia, with a long course and high probability of relapse and prolongation. Long-term complications caused by long-term usage of hormones and immunosuppressants in children with INS seriously affect their physical and mental health and quality of life. Most children with steroid-sensitive nephrotic syndrome can be cured before adulthood, while some of them relapse in adulthood. Long-term prognosis of children with steroid-resistant nephrotic syndrome is poor. There have been few studies in China followed the long-term outcomes and its related factors of children with INS over 10 years. The paper reviewed the literatures on the long-term outcomes of children with INS, including renal survival, growth, mental health, learning and work, marriage and fertility, disease recurrence and long-term related complications, to explore the factors related to the poor long-term outcomes of children with INS and to assist in clinical decision-making and follow-up management.