Objective:To investigate the efficacy and safety of Rituximab (RTX) in the treatment of children with frequently relapsing/steroid-dependent nephrotic syndrome (FRNS/SDNS) and to analyze the factors influencing the efficacy.Methods:Case series study.The clinical data of children with FRNS/SDNS who received B-cell-guided RTX (single dose: 375 mg/m 2, maximum dose: 500 mg, one additional dose when peripheral blood CD19 + B lymphocytes ≥0.01) in the First Affiliated Hospital of Zhengzhou University from September 2019 to March 2022 were retrospectively collected.The frequency of relapse and cumulative dose of glucocorticoids before and after RTX treatment were compared.The Kaplan-Meier method was used to analyze relapse-free survival rate and FRNS/SDNS-free survival rate after RTX treatment.The influencing factors of relapse were analyzed using the Cox proportional hazards regression model. Results:Totally 47 children were enrolled, including 35 males and 12 females; the age of first application of RTX was 10.2 (6.9, 13.0) years; 33 children had used one type of immunosuppressant before, and 14 children had used two or more types of immunosuppressant before; the dose of RTX treatment was 3.0 (2.0, 3.0). The frequency of relapse[0(0, 0.55) times/year vs.1.62 (1.09, 2.40) times/year] and cumulative dose of glucocorticoids[0.12 (0.05, 0.21) mg/(kg·d) vs.0.40 (0.20, 0.56) mg/(kg·d)] after RTX treatment significantly decreased compared with previous immunosuppressive treatment ( Z=-5.56, -5.54, all P<0.001). The relapse-free survival rates at 6, 12, 18 and 24 months after treatment were 80.9%, 72.3%, 68.1% and 68.1%, respectively, and the FRNS/SDNS-free survival rates were 93.6%, 89.4%, 89.4% and 89.4%, respectively.Univariate Cox regression analysis showed that the high frequency of relapse during previous immunosuppressive therapy was a risk factor for relapse after RTX treatment ( P<0.05). Of the 14 children who relapsed, 6 occurred in children whose CD19 + B lymphocytes<0.01, and the frequency of relapse after RTX treatment was significantly higher than those whose CD19 + B lymphocytes≥0.01 ( Z=-2.84, P=0.005). No severe adverse reactions occurred during RTX treatment and follow-up. Conclusions:The B-cell-guided RTX is effective and safe in the treatment of FRNS/SDNS in children.The high frequency of relapse during previous immunosuppressive therapy is a risk factor for relapse after RTX treatment, and relapse in the state of B lymphocyte depletion predicts poor outcomes of RTX treatment.

β-glucosidase has important applications in food, pharmaceutics, biomass conversion and other fields, exploring β-glucosidase with strong adaptability and excellent properties thus has received extensive interest. In this study, a novel glucosidase from the GH1 family derived from Cuniculiplasma divulgatum was cloned, expressed, and characterized, aiming to find a better β-glucosidase. The amino acid sequences of GH1 family glucosidase derived from C. divulgatum were obtained from the NCBI database, and a recombinant plasmid pET-30a(+)-CdBglA was constructed. The recombinant protein was induced to express in Escherichia coli BL21(DE3). The enzymatic properties of the purified CdBglA were studied. The molecular weight of the recombinant CdBglA was 56.0 kDa. The optimum pH and temperature were 5.5 and 55 ℃, respectively. The enzyme showed good pH stability, 92.33% of the initial activity could be retained when treated under pH 5.5-11.0 for 1 h. When pNPG was used as a substrate, the kinetic parameters K_m, V_max and K_cat/K_m were 0.81 mmol, 291.99 μmol/(mg·min), and 387.50 s^-1 mmol^-1, respectively. 90.33% of the initial enzyme activity could be retained when CdBglA was placed with various heavy metal ions at a final concentration of 5 mmol/L. The enzyme activity was increased by 28.67% under 15% ethanol solution, remained unchanged under 20% ethanol, and 43.68% of the enzyme activity could still be retained under 30% ethanol. The enzyme has an obvious activation effect at 0-1.5 mol/L NaCl and can tolerate 0.8 mol/L glucose. In conclusion, CdBglA is an acidic and mesophilic enzyme with broad pH stability and strong tolerance to most metal ions, organic solvents, NaCl and glucose. These characteristics may facilitate future theoretical research and industrial production.
beta-Glucosidase ; Sodium Chloride ; Temperature ; Glucose ; Ethanol/chemistry* ; Ions ; Hydrogen-Ion Concentration ; Enzyme Stability ; Substrate Specificity

Objective:To observe the correlation between homocysteine (Hcy) and serum uric acid (SUA) and retinopathy in type 2 diabetes mellitus (T2DM), preliminary study on its predictive value.Methods:A retrospective study. From January 2020 to March 2021, a total of 324 T2DM patients hospitalized in Department of Endocrinology, Cangzhou Central Hospital of Hebei Province were included. Fasting blood glucose (FBG), glycated hemoglobin (HbA1C), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), serum creatinine (Scr), blood urea nitrogen (BUN), Hcy, SUA, peripheral blood endothelial progenitor cells (EPC), circulating endothelial cells (CEC) were counted and homeostasis model assessment for insulin resistance (HOMA-IR) was calculated. According to the absence or presence of diabetic retinopathy (DR), the patients were divided into non DR (NDR) group and DR group with 100 and 214 cases, respectively. Clinical data and laboratory biochemical indexes of the two groups were compared and observed. The logistic regression was used to analyze the independent risk factors for DR in T2DM patients. Smooth curve fitting was used to analyze the curve relationship between Hcy, SUA and DR, and ROC area (AUC) of Hcy, SUA; their combined prediction of DR in T2DM patients was calculated by receiver operating characteristic curve (ROC curve), and the predictive value of Hcy and SUA for DR in T2DM patients was evaluated.Results:Diabetic course ( t=5.380), systolic blood pressure ( t=2.935), hypertension ( χ2=10.248), diabetic nephropathy ( χ2=9.515), diabetic peripheral neuropathy ( χ2=24.501), FBG ( t=3.945), HbA1C ( t=3.336) and TG in DR Group ( t=2.898), LDL-C ( t=3.986), Scr ( t=2.139), SUA ( t=7.138), HOMA-IR ( t=3.237), BUN ( t=3.609), Hcy ( t=2.363) and CEC ( t=19.396) were significantly higher than those in NDR group. The difference was statistically significant ( P<0.05). EPC ( t=9.563) and CPC ( t=7.684) levels were significantly lower than those of NDR group, and the difference was statistically significant ( P<0.05). Logistic regression analysis showed that diabetes course, SBP, hypertension, FBG, HbA1C, LDL-C, SUA, Hcy, EPC, CPC and CEC were all independent risk factors for developing DR in T2DM patients ( P<0.05). The smooth curve fitting analysis showed that Hcy and SUA were positively correlated with the occurrence of DR. After adjusting for confounding factors, when Hcy≥15 μmol/L, the risk of DR Increased by 14% for every 1 μmol/L increase in Hcy [odds ratio ( OR)=0.92, 95% confidence interval ( CI) 0.88-0.98, P<0.05]. When Hcy <15 μmol/L, there was no significant difference ( OR=0.96, 95% CI 0.92-1.08, P>0.05). When SUA≥304 μmol/L, the risk of DR increased by 17%, every 20 μmol/L SUA increased ( OR=0.80, 95% CI 0.68-0.94, P<0.05). When SUA <304 μmol/L, the difference was not statistically significant ( OR=0.83, 95% CI 0.72-0.95, P>0.05). ROC curve analysis results showed that the AUC values of Hcy, SUA and Hcy combined with SUA in predicting the occurrence of DR in T2DM patients were 0.775 (95% CI 0.713-0.837, P<0.001), 0.757 (95% CI 0.680-0.834, P<0.001) and 0.827 (95% CI 0.786-0.868, P<0.001). Hcy combined with SUA showed better predictive efficiency. Conclusions:The abnormal increase of Hcy and SUA levels in T2DM patients are closely related to the occurrence of DR, they are independent risk factors for the occurrence of DR. Hcy combined with SUA has high predictive value for the occurrence of DR.

Metabolic dysfunction-associated fatty liver disease is becoming the most common cause of chronic liver disease worldwide, with a disease spectrum including simple steatosis, steatohepatitis, hepatic fibrosis/cirrhosis, and liver cancer. Most metabolic dysfunction-associated fatty liver diseases progress slowly, but steatohepatitis, especially in patients accompanied by significant liver fibrosis, has a significantly increased risk of adverse liver disease outcomes and all-cause death. Therefore, early-stage identification of medium-and high-risk groups carried out by stratified management has important clinical significance. Pathological diagnosis is the gold standard for diagnosing steatohepatitis and liver fibrosis. However, its invasiveness, sampling errors, and unsuitability for dynamic monitoring limit its clinical application. In recent years, a large number of non-invasive diagnostic methods based on somatology, serology, and imaging have shown great development prospects in order to meet the clinical needs of assessing disease severity and risk stratification. This article reviews and summarizes the application and progress of magnetic resonance imaging technology in the non-invasive diagnosis of metabolic dysfunction-associated fatty liver disease.

β-glucosidase has important applications in food, medicine, biomass conversion and other fields. Therefore, exploring β-glucosidase with strong stability and excellent properties is a research hotspot. In this study, a GH3 family β-glucosidase gene named Iubgl3 was successfully cloned from Infirmifilum uzonense. Sequence analysis showed that the full length of Iubgl3 was 2 106 bp, encoding 702 amino acids, with a theoretical molecular weight of 77.0 kDa. The gene was cloned and expressed in E. coli and the enzymatic properties of purified IuBgl3 were studied. The results showed that the optimal pH and temperature for pNPG hydrolysis were 5.0 and 85 ℃, respectively. The enzyme has good thermal stability, and more than 85% of enzyme activity can be retained after being treated at 80 ℃ for2 h. This enzyme has good pH stability and more than 85% of its activity can be retained after being treated at pH 4.0-11.0 for 1 h. It was found that the enzyme had high hydrolysis ability to p-nitrophenyl β-d-glucoside (pNPG) and p-nitrophenyl β-d-xylopyranoside (pNPX). When pNPG was used as the substrate, the kinetic parameters K_m and V_max were 0.38 mmol and 248.55 μmol/(mg·min), respectively, and the catalytic efficiency k_cat/K_m was 6 149.20 s^-1mmol^-1. Most metal ions had no significant effect on the enzyme activity of IuBgl3. SDS completely inactivated the enzyme, while EDTA increased the enzyme activity by 30%. This study expanded the β-glucosidase gene diversity of the thermophilic archaea GH3 family and obtained a thermostable acid bifunctional enzyme with good industrial application potential.
beta-Glucosidase/chemistry* ; Archaea/metabolism* ; Escherichia coli/metabolism* ; Hydrogen-Ion Concentration ; Temperature ; Glucosides ; Enzyme Stability ; Substrate Specificity ; Kinetics

Objective:To summarize the clinical features and gene phenotype of children with spondyloenchondrodysplasia with immune dysregulation (SPENCDI) caused by ACP5 gene mutation. Methods:The medical data and genetic phenotype of a child diagnosed with SPENCDI in the Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University in February 23, 2017 were analyzed retrospectively.Besides, " spondyloenchondrodysplasia" were taken as the search terms to perform the retrieval in CNKI, Wanfang Data, and PubMed, in an attempt to conduct the literature review.χ 2 test was used to compare the factors among children with different mutations. Results:The 4.5-year-old girl was admitted to hospital for complaint of " fever and chilblain-like rash" when she was 2 years old.She was diagnosed with systemic lupus erythematosus (SLE) concomitant with lupus nephritis.Methylprednisolone combined with cyclophosphamide, mycophenolate mofetil was used for the treatment.However, she experienced multiple infections, thrombocytopenia, limp, and growth retardation during the treatment.Genetic detection identified ACP5 gene compound hybrid mutation: c.779C>A and c. 770T>C.She was diagnosed with SPENCDI, and was subjected to follow-up.A total of 78 SPENCDI patients were retrieved from the databases, with various clinical manifestations of SPENCDI, commonly with skeletal involvement and immune phenotypes; 73.08% of the cases were positive for antinuclear antibodies, 57.69% of cases were positive for anti-double stranded-DNA antibodies and 34.62% of cases had neurological symptoms.In 58 cases, ACP5 gene mutations were detected, including 44 homozygous mutations and 14 compound heterozygous mutations.Patients with ACP5 gene homozygous mutation had a higher probability of consanguineous marriage in parents [56.82% (25/44 cases) vs.14.29% (2/14 cases)]; patients with ACP5 gene heterozygous mutation were more likely to develop SLE [64.29% (9/14 cases) vs.34.09% (15/44 cases)]( χ2=7.722, 3.992; all P<0.05). Conclusions:The majority of the ACP5 gene mutations are homozygous mutations in patients with SPENCDI, and heterozygous mutations are rare.The clinical manifestations of SPENCDI are various and complex, it is prone to develop autoimmune diseases, and there was no clear correlation between clinical features and gene phenotype in SPENCDI patients.

Objective:To construct an index system for evaluating the development of research-oriented disciplines in medical institutions, and to provide a basis for guiding the superior disciplines towards international high-level disciplines.Methods:Through literature research and expert discussions, the basic framework of the research-oriented disciplines development evaluation system was established. The final version of the index system was determined using the Delphi method and expert interviews, and the weights of each index were established using the hierarchical analysis method.Results:After consultation, the framework of 5 first-class indexes, 16 second-class indexes, and 51 third-class indexes was finally constructed for the evaluation system of research-oriented discipline development, and the weights of indexes at different classes were calculated. The high-frequency terms " research mechanism construction" , " research team construction" and " representative research results" extracted from the expert interviews echoed the results of the weighting calculation.Conclusions:The evaluation system for research-oriented disciplines constructed in this study is relatively reasonable, scientific, reliable, and operable. The construction of the index system adapts to the characteristics of the development of research-oriented disciplines, with guiding significance for enhancing the development of research-oriented disciplines.

Objective:To explore the effect of a Chinese medicinal composition ( Xiadanqi) on the prevention of radon exposure induced injuries of lung in vitro and in vivo. Methods:Mice were randomly divided into three groups of blank control group, radon-exposed group alone and radon-exposed group intervened with Chinese medicinal composition. The pathological changes of lung tissues in each group after 120 WLM were observed by HE and Masson staining, and the expressions of α-SMA protein and Vimentin protein in lung tissues were detected by immunohistochemistry staining. The levels of oxidative stress in lung tissue of each group were detected with SOD and MDA kits. At the same time, a radon exposed cell model and a radon exposure + Xiadanqi intervention cell model were constructed using an ecological radon chamber. The cell adhesion abilities of different groups were detected by an adhesion kit. The cell migration ability of each group was determined by the transwell migration experiment. The expression of E-cadherin and Vimentin protein was detected by Western blot. Results:Compared with the radon exposure group, the concentration of MDA was decreased ( t=4.43, P<0.05), the activity of SOD was increased ( t=3.22, P<0.05), and α-SMA and Vimentin protein expressions were decreased ( t=3.08, 7.57, P<0.05) in lung tissue of mice intervened with 2 mg/g Xiadanqi. In vitro, compared with radon exposure group, the migration ability was reduced ( t=4.78, 13.01, P<0.05), the cell adhesion property was enhanced ( t=3.41, 12.55, P<0.05), the expression of E-cadherin protein was increased ( t=2.96, 19.57, P<0.05), and the expression of Vimentin protein was obviously reduced ( t=21.00, 33.32, P<0.05) in radon-exposed cells with the treatment of Chinese medicine (150 μg/ml and 200 μg/ml). Conclusions:The Chinese medicinal composition ( Xiadanqi) has a certain radioprotective effect on radon exposure induced injury by reducing oxidative stress, attenuating EMT and fibrosis, and thus it may be applied as a protective agent for radon induced injury.

Objective To investigate the outcomes of patients with diabetic retinopathy (DR) in Shanghai communities and the influencing factors.Methods From October 2015 to April 2016,533 type 2 diabetic patients with DR were selected by target sampling and cluster random sampling method from six community health service centers in Shanghai.Patients were followed up for 1 year.The demographic information,physical examination,laboratory tests and eye fundus exam results were documented and the DR was graded.The factors associated with the regression of DR were analyzed.Results Total 478 patients,including 280 females (58.6％) and 198 males (41.4％),were followed up for 1 year.The mean age of patients was (64±7) years and the mean disease duration was (8.85±4.20) years.The original DR lesion was remitted in 35 patients with an improvement rate of 7.3％;while the original DR lesion was aggravated in 29 patients with a progression rate of 6.1％.Ordinal logistic regression analysis revealed that age (OR=0.197,95％CI:0.056-0.699),body mass index (BMI) (OR=0.383,95％CI:0.171-0.856),glycosylated hemoglobin (HbAlc) (OR=0.287,95％CI:0.102-0.803),triglycerides (TG) (OR=0.541,95％CI:0.295-0.991),urinary albumin to creatinine ratio (ACR)(OR=0.218,95％CI:0.066-0.720) were associated with DR in type 2 diabetic patients.Conclusion The regression of DR is closely related to age,BMI,glucose,serum lipids and renal function,so it is suggested that lowering BMI,controlling glucose and serum lipids and maintaining normal kidney function are necessary for preventing the progression and promoting the improvement of DR in diabetic patients.

Objective To explore the clinical features,auxiliary examinations,therapies and prognoses of patients with antibodies against contactin-associated protein-like 2 (CASPR2).Methods The clinical data of 11 anti-CASPR2 encephalitis patients who were admited to the People's Hospital of Zhengzhou University from March 2015 to April 2018 were retrospectively analyzed.Results The age of these 11 cases was (35.6± 19.4) years (ranged 20-74 years),and eight cases were females.There were seven cases with limbic encephalitis which included six cases of epilepsy,four cases of memory impairment,two cases of mental and behavioral abnormalities.Four cases had peripheral nerve hyperexcitability.Four cases had neuropathic pain.There were six cases with autonomic dysfunction including five cases of constipation,three cases of tachycardia,two cases of hyperhidrosis,two cases of urinary disorder.Seven cases had sleep disorder.Four cases had weight loss.Two cases showed cerebellar symptoms and two cases had hyponatremia.Magnetic resonance imaging scan of the brain showed abnormal signal in two cases,mainly involved medial temporal lobe and the hippocampus.Six cases underwent 18F-fluorodeoxyglucose positron emission tomography-computed tomography (PET-CT) examination,and three cases showed abnormalities,including two with temporal hypermetabolism and one with cortical hypermetabolism.Chest enhanced CT and PET-CT showed thymoma in one case.All cases received immunotherapy,and after treatment their symptoms were improved.Long-term follow-up was performed in nine cases,and three cases relapsed.Conclusions The major clinical manifestations of anti-CASPR2 encephalitis were limbic encephalitis,peripheral nerve hyperexcitability,neuropathic pain,autonomic dysfunction,insomnia and so on.Immunotherapy was effective and some patients may have recurrence.