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

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 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.

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 summarize the risk factors, clinical manifestations, diagnosis and treatment of limb thrombosis in neonates.Methods:The clinical data of 14 neonates with limb thrombosis were hospitalized in neonatology department at Shengjing Hospital of China Medical University from February 2012 to February 2022 were retrospectively analyzed.Results:All the 14 cases of limb thrombosis were premature infants, with an average gestational age of 29 weeks and 5 days(27 weeks and 3 days to 33 weeks and 1 day), including eight cases of arterial embolism and six cases of venous embolism.Among them, 13(92.9%) cases were diagnosed with infectious diseases such as septicemia or neonatal necrotizing enterocolitis within 48 hours before embolization, and all had a history of peripheral arterial and venous catheterization.During the early stage of embolization, limb artery embolism was characterized by weakened distal artery pulsation, pale skin, gradual cyanosis and even gangrene.Limb venous embolism was manifested as limb swelling, skin congestion and cyanosis, but the arterial pulsation was normal.Fourteen cases were confirmed by vascular ultrasound.All the eight cases with arterial embolization were treated with heparin anticoagulation, five of which were cured, with an effective rate of 62.5%.The average time of heparin use in five cases was 2.5 days(2-3 days). One patient was not effective after 2 days of heparin treatment, and recovered after thrombectomy.Another two cases had distal limb gangrene, and them were treated with heparin for 5 days and 7 days.All of the six cases with venous embolism were cured, of which four cases were treated with heparin for an average of 8.5 days(4-15 days), and the other two cases were cured after general treatment.There were no bleeding events in the 12 infants treated with heparin.Conclusion:Peripheral arterial and venous catheterization during infection of preterm infants is the most common cause of limb thrombosis.The smaller body weight and gestational age, the thinner blood vessels, the higher risk of occurrence.Vascular ultrasound is the most commonly used examination method for neonatal thrombosis, and heparin anticoagulant therapy is the most commonly used treatment measure.When the treatment effect of heparin is not good, other treatment methods should be sought.

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.

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.

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 explore the clinical characteristics and risk factors of neonatal adrenal hemorrhage(NAH), and to improve the understanding, diagnosis and treatment of this disease.Methods:In this study, a retrospective nested case-control study was used to collect clinical data of neonates diagnosed with NAH from January 2011 to December 2021 in the Department of Neonatology, Shengjing Hospital of China Medical University, and telephone follow-up were conducted for them.NAH infants with manifestations of neonatal hyperbilirubinemia were selected as the case group, and the random number table method was used to select the neonatal hyperbilirubinemia infants with NAH excluded by imaging in the same period at a ratio of 1: 2 as the control group.Characteristics of the clinical data of the two groups were compared and analyzed by Logistic regression to explore the risk factors of NAH.Results:During the study period, a total of 31 cases of NAH were diagnosed, with an average gestational age of(37.6±2.2) weeks, including 19 males, 25 full-term infants, 6 cases with macrosomia, 30 cases with natural labor, 29 cases with hyperbilirubinemia, 8 cases with birth injury, 7 cases with asphyxia, 9 cases with bilirubin encephalopathy, 12 cases with sepsis, 13 cases with intracranial hemorrhage, 17 cases with anemia, 9 cases with respiratory disease, 5 cases with hyperkalemia, 6 cases with hyponatremia.The results of NAH ultrasonography showed that 8 cases of hematoma had medium and low echoes, 6 cases of mixed echoes, and 17 cases of liquid flocculent echoes with or without punctate echoes.Color Doppler flow imaging results showed no blood flow signal.There were 26 cases on the right side, 4 cases on the left side, and 1 case on both sides.A total of 26 cases were followed up.Ultrasonography showed that most haematomas were absorbed within 1 to 3 months and disappeared within 6 months.Twenty-nine cases were included in the case group and 58 cases in the control group.Univariate analysis showed that age, birth weight, macrosomia, mode of delivery, bilirubin encephalopathy, neonatal sepsis, abdominal distension, anemia, asphyxia, total bilirubin, indirect bilirubin, Hb and CRP were significantly higher than those in the control group( P<0.05). Multivariate logistic regression analysis showed that macrosomia( OR=7.415, 95% CI=1.342~40.956, P=0.022) and asphyxia( OR=12.075, 95% CI=1.293~112.736, P=0.029) were independent risk factors of NAH. Conclusion:NAH is common in naturally born full-term infants, with a lack of specific clinical manifestations.Unexplained persistent hyperbilirubinemia may be its first symptom, often accompanied by anemia and ion disturbance.A few infants may have adrenal insufficiency.Macrosomia and asphyxia may be the risk factors for the occurrence of NAH.