Simultaneous determination of 18 common disinfection by-product residues in drinking water by high performance liquid chromatography-tandem mass spectrometry / 上海预防医学

ObjectiveA method was developed for the rapid determination of 18 common disinfection by-products including halogenated oxides and haloacetic acid （HAAs） in drinking water by high performance liquid chromatography tandem mass spectrometry （HPLC-MS/MS）. MethodThe water sample was filtered by 0.22 μm hydrophilic membrane then the analytes were separated on a PFP （2.1 mm× 100 mm， 2.7 μm） pentafluorophenyl column with 0.1% acetic acid and acetonitrile as mobile phase gradient elution. Ionization in anionic electrospray mode was detected by multi-reaction monitoring （MRM） mode. The external standard method was used for quantitation. ResultsThe correlation coefficients of 18 disinfection by-products were above 0.999 in the corresponding linear range. The average spiked recoveries of 1， 10 and20 times of LOQ of each analyte were 91.6%‒101.8%， and the relative standard deviation （RSD） was 1.2%‒6.4%. The LOD and LOQ were 0.020‒2 μg·L-1 and 0.050‒5 μg·L-1， respectively. ConclusionThis method is simple， sensitive and accurate， and could be used for the routine analysis of 18 common disinfection by-products in drinking water.

A Retrospective Study on the Efficacy and Safety of Idarubicin Combined with High-Dose Cytarabine Consolidation in Patients with Acute Myeloid Leukemia over 60 Years of Age in First Remission / 中国实验血液学杂志

OBJECTIVE@#To evaluate the efficacy and safety of idarubicin combined with high-dose cytarabine as a post-remission therapy for elderly patients with acute myeloid leukemia (AML).@*METHODS@#From November 2017 to June 2021, 24 AML patients aged ≥60 years who were in complete remission for the first time were enrolled in consolidation chemotherapy with idarubicin (10 mg/m2 intravenously once for day 1) combined with high-dose cytarabine (1.5 g/m2 intravenously over 3 hours every 12 hours for day 1-3), and the efficacy and safety were observed.@*RESULTS@#Among the 24 patients, there were 12 males and 12 females, the median age was 65 (60-78) years old, and the median follow-up time was 23.3 (2-42.7) months. By the end of the follow-up, 15 patients relapsed and 11 patients died. The median disease-free survival (DFS) was 9 months and there were 3 cases of 2-year DFS. The median overall survival (OS) was 16.2 months, and there were 4 cases of 2-year OS. In terms of safety, 6 patients had grade 1-2 non-hematological adverse reactions, 12 patients had grade 3-4 hematological adverse reactions, and a total of 6 patients developed infection after consolidation chemotherapy. Multivariate analysis showed that two induction cycles and high-risk cytogenetic abnormalities were the adverse factors of DFS and OS in elderly patients with AML in this study.@*CONCLUSION@#For AML patients ≥60 years old in first complete remission, idarubicin combined with high-dose cytarabine as post-remission therapy has a better safety, but compared with other regimens does not improve the prognosis of elderly patients, which needs further exploration.

Clinical application of three-dimensional printed preformed titanium mesh combined with free latissimus dorsi muscle flap in the treatment of squamous cell carcinoma with skull defect in the vertex / 中华烧伤杂志

Objective: To explore the clinical effects of three-dimensional printed preformed titanium mesh combined with latissimus dorsi muscle flap free transplantation in the treatment of wounds with skull defect after radical surgery of squamous cell carcinoma in the vertex. Methods: A retrospective observational study was conducted. From January 2010 to December 2019, 5 patients with squamous cell carcinoma in the vertex accompanied with skull invasion who met the inclusion criteria were admitted to the Department of Burns and Plastic Surgery of the Second Affiliated Hospital of Air Force Medical University, including four males and one female, aged 50 to 65 years. The original lesion areas ranged from 5 cm×4 cm to 15 cm×8 cm. The titanium mesh was prefabricated via three-dimensional technic based on the result the scope of skull resection predicted with computerized tomography three-dimensional reconstruction before surgery. During the first stage, the soft tissue defect area of scalp (8 cm×7 cm to 18 cm×11 cm) after tumor enlargement resection was repaired with the preformed titanium mesh, and the titanium mesh was covered with latissimus dorsi muscle flap, with area of 10 cm×9 cm to 20 cm×13 cm. The thoracodorsal artery/vein was anastomosed with the superficial temporal artery/vein on one side. The muscle ends in the donor site were sutured together or performed with transfixion, and then the skin on the back were covered back to the donor site. On the 10th day after the first-stage surgery, the second-stage surgery was performed. The thin intermediate thickness skin graft was taken from the anterolateral thigh to cover the latissimus dorsi muscle flap. The duration and intraoperative blood loss of first-stage surgery were recorded. The postoperative muscle flap survival after the first-stage surgery and skin graft survival after the second-stage surgery was observed. The occurrence of complications, head appearance, and recurrence of tumor were followed up. Results: The average first-stage surgery duration of patients was 12.1 h, and the intraoperative blood loss was not more than 1 200 mL. The muscle flaps in the first-stage surgery and the skin grafts in the second-stage surgery all survived well. During the follow-up of 6-18 months, no complications such as exposure of titanium mesh or infection occurred, with good shape in the recipient sites in the vertex, and no recurrence of tumor. Conclusions: Three-dimensional printed preformed titanium mesh combined with latissimus dorsi muscle flap free transplantation and intermediate thickness skin graft cover is an effective and reliable method for repairing the wound with skull defect after extended resection of squamous cell carcinoma in the vertex. This method can cover the wound effectively as well as promote both recipient and donor sites to obtain good function and appearance.

Association of blood lead and blood selenium with serum high-sensitivity C-reactive protein among Chinese adults aged 19 to 79 years / 中华流行病学杂志

Objective: To investigate the association of blood lead and blood selenium with serum high-sensitivity C-reactive protein (hs-CRP) among Chinese adults aged 19 to 79 years. Methods: The participants were enrolled from the first wave of China National Human Biomonitoring (CNHBM) conducted from 2017 to 2018. 10 153 participants aged 19 to 79 years were included in this study. Fasting blood samples were obtained from participants. Lead and selenium in whole blood and hs-CRP in serum were measured. Individuals with hs-CRP levels above 3.0 mg/L were defined as elevated hs-CRP. Generalized linear mixed models and restricted cubic spline models were used to analyze the association of blood lead and blood selenium with elevated hs-CRP. Logistic regression models were used to analyze the multiplicative scale and additive scale interaction between blood lead and blood selenium on elevated hs-CRP. Results: The age of participants was (48.91±15.38) years, of which 5 054 (61.47%) were male. 1 181 (11.29%) participants were defined as elevated hs-CRP. After multivariable adjustment, results from generalized linear models showed that compared with participants with the lowest quartile of blood lead, the OR (95%CI) of elevated hs-CRP for participants with the second, third, and highest quartiles were 1.14 (0.94-1.37), 1.25 (1.04-1.52) and 1.38 (1.13-1.68), respectively. When compared with participants with the lowest quartile of blood selenium, the OR (95%CI) of elevated hs-CRP for participants with the second, third and highest quartiles were 0.86 (0.72-1.04), 0.91 (0.76-1.11), and 0.75 (0.61-0.92), respectively. Results from the interaction analysis showed no significant interaction between lead and selenium on elevated hs-CRP. Conclusion: Blood concentration of lead was positively associated with elevated serum hs-CRP, and blood concentration of selenium was inversely related to elevated hs-CRP, while blood lead and selenium did not present interaction on elevated hs-CRP.

Psychological and behavioral problems in children and adolescents during the coronavirus disease 2019 epidemic: a Scoping review / 中国当代儿科杂志

OBJECTIVES@#To investigate the psychological and behavioral problems and related influencing factors in children and adolescents during the coronavirus disease 2019 (COVID-19) epidemic.@*METHODS@#China National Knowledge Infrastructure, Wanfang Data, PubMed, and Web of Science were searched using the method of subject search for articles published up to March 31, 2022, and related data were extracted for Scoping review.@*RESULTS@#A total of 3 951 articles were retrieved, and 35 articles from 12 countries were finally included. Most of the articles were from the journals related to pediatrics, psychiatry, psychology, and epidemiology, and cross-sectional survey was the most commonly used research method. Psychological and behavioral problems in children and adolescents mainly included depression/anxiety/stress, sleep disorder, internet behavior problems, traumatic stress disorder, and self-injury/suicide. Influencing factors were analyzed from the three aspects of socio-demographic characteristics, changes in living habits, and ways of coping with COVID-19.@*CONCLUSIONS@#During the COVID-19 epidemic, the psychological and behavioral problems of children and adolescents in China and overseas are severe. In the future, further investigation and research can be carried out based on relevant influencing factors to improve the psychological and behavioral problems.

Influencing factors for the quality of bowel preparation with polyethylene glycol electrolyte powder combined with diet control before colonoscopy in children / 中国当代儿科杂志

OBJECTIVES@#To investigate the influencing factors for the quality of bowel preparation before colonoscopy in children and the association of the interval from the last administration of laxative to the start of colonoscopy (shortly referred to as waiting time) with the quality of bowel preparation.@*METHODS@#A retrospective analysis was performed for the children who were admitted to the Department of Gastroenterology, Children's Hospital of Nanjing Medical University, from January to November 2020, and received bowel preparation with polyethylene glycol electrolyte powder combined with diet control before colonoscopy. According to the score of Boston bowel preparation scale, they were divided into two groups: adequate bowel preparation group (n=337) and inadequate bowel preparation group (n=30). Related data were collected from the children in both groups, including general information, possible influencing factors for the quality of bowel preparation, adverse reactions associated with bowel preparation, duration of colonoscopy, and postoperative diagnosis. Univariate and multivariate analyses were used to explore the influencing factors for the quality of bowel preparation.@*RESULTS@#The univariate analysis showed that age, body weight, and waiting time were associated with inadequate bowel preparation (P<0.05). The multivariate analysis showed that older age (OR=2.155, 95%CI: 1.087-4.273, P=0.028) and longer waiting time (OR=1.559, 95% CI: 1.191-2.041, P=0.001) were independent risk factors for inadequate bowel preparation. The receiver operating characteristic (ROC) curve analysis showed that the cut-off value of waiting time was 5.5 hours in determining whether bowel preparation was adequate or not, with a sensitivity of 90.0%, a specificity of 50.7%, and an area under the ROC curve of 0.708. After grouping based on waiting time, it was found that the incidence rate of inadequate bowel preparation in the ≥5.5 hours group was significantly higher than that in the <5.5 hours group [14.0% (27/193) vs 1.7% (3/174), P<0.001].@*CONCLUSIONS@#For children who use polyethylene glycol electrolyte powder combined with diet control for bowel preparation, older age is an independent risk factor for inadequate bowel preparation before colonoscopy, which may be associated with an insufficient dose of polyethylene glycol in older children. Longer waiting time is also an independent risk factor for inadequate bowel preparation, and it is recommended that the waiting time should not exceed 5.5 hours.

Long Noncoding RNA PVT1 Promotes Stemness and Temozolomide Resistance through miR-365/ELF4/SOX2 Axis in Glioma

Long non-coding RNA (lncRNA) are a class of non-coding RNAs demonstrated to play pivotal roles in regulating tumor progression. Therefore, deciphering the regulatory role of lncRNA in the development of glioma may offer a promising therapeutic target for treatment of glioma. We performed RT-qPCR analysis on the expression of lncRNA plasmacytoma variant translocation 1 (PVT1) and miR-365 in glioma tissues and cell lines. Cell proliferation and viability was assessed with CCK8 assay. Cell migration was assessed by wound healing assay. Transwell assay was used to assess cell invasion capacity. Expression of CD133+ cells was detected by flow cytometry. Western blot assay was used to detection the expression of ELF4 and stemness-related protein SOX2, Oct4 and Nanog. Bioinformatics and dual-luciferase assay were used to predict and validate the interaction between PVT1 and miR-365. Elevated PVT1 expression was observed in glioma tissues and cells. Knockdown of PVT1 and overexpression of miR-365 inhibited proliferation, migration, invasion and promoted stemness and Temozolomide (TMZ) resistance of glioma cells. PVT1 regulated ELF4 expression by competitively binds to miR-365. PVT1 regulated the stemness and sensitivity of TMZ of glioma cells through miR-365/ELF4/ SOX2 axis. This study identified that PVT1 promoted glioma stemness through miR-365/ELF4/SOX2 axis.

Effect of acupuncture on the cognitive function of migraine patients with depression/anxiety disorder / 中国针灸

OBJECTIVE@#To observe the effect of five-element acupuncture on the cognitive function repair of migraine patients with depression/anxiety disorder.@*METHODS@#The migraine patients with depression/anxiety disorder (19 cases, 5 cases dropped off) were taken as the observation group, and received five-element acupuncture twice a week for 8 weeks. Healthy subjects (19 cases) were selected by demographic data matching as the control group. The cognitive function was evaluated with the event related potential (ERP) technique, and the latency and amplitude of visual evoked potential P300 were adopted as the observation indexes. The headache days (every 4 weeks), headache intensity [visual analogue scale(VAS) score], and headache impact test-6 (HIT-6) score, Hamilton depression scale (HAMD) score and Hamilton anxiety scale (HAMA) score were used as the observation indexes for curative effect.@*RESULTS@#Before the treatment, latency of target stimulus at Fz [ (417.5±34.3) ms] in the observation group was extended compared with the healthy subjects of the control group [(388.6±42.1) ms, @*CONCLUSION@#There are some cognitive impairments in migraine patients with depression/anxiety disorder. Five-element acupuncture not only relieves headache, anxiety and depression effectively, but also improves the activation level of the frontal lobe. It significantly repairs the impaired cognitive function.

Associations of Sarcopenia, Handgrip Strength and Calf Circumference with Cognitive Impairment among Chinese Older Adults / 生物医学与环境科学（英文）

Objective@#To evaluate the associations of sarcopenia, handgrip strength and calf circumference with cognitive impairment among Chinese older adults.@*Methods@#Totally 2,525 older adults were recruited from the Healthy Aging and Biomarkers Cohort Study. Cognitive impairment was assessed by the Chinese Mini-Mental State Examination. Handgrip strength was calculated from the means of the right and left hand values. Calf circumference was measured at the site of maximum circumference of the non-dominant leg. The formula developed by Ishii was used to define sarcopenia. Multiple logistic regression was performed to evaluate the associations of sarcopenia, handgrip strength, and calf circumference with cognitive impairment.@*Results@#The prevalence of cognitive impairment was 34.36%. The adjusted odds ratio ( @*Conclusion@#Sarcopenia, identified by low handgrip strength and low calf circumference, was positively associated with cognitive impairment.

Long Noncoding RNA PVT1 Promotes Stemness and Temozolomide Resistance through miR-365/ELF4/SOX2 Axis in Glioma

Long non-coding RNA (lncRNA) are a class of non-coding RNAs demonstrated to play pivotal roles in regulating tumor progression. Therefore, deciphering the regulatory role of lncRNA in the development of glioma may offer a promising therapeutic target for treatment of glioma. We performed RT-qPCR analysis on the expression of lncRNA plasmacytoma variant translocation 1 (PVT1) and miR-365 in glioma tissues and cell lines. Cell proliferation and viability was assessed with CCK8 assay. Cell migration was assessed by wound healing assay. Transwell assay was used to assess cell invasion capacity. Expression of CD133+ cells was detected by flow cytometry. Western blot assay was used to detection the expression of ELF4 and stemness-related protein SOX2, Oct4 and Nanog. Bioinformatics and dual-luciferase assay were used to predict and validate the interaction between PVT1 and miR-365. Elevated PVT1 expression was observed in glioma tissues and cells. Knockdown of PVT1 and overexpression of miR-365 inhibited proliferation, migration, invasion and promoted stemness and Temozolomide (TMZ) resistance of glioma cells. PVT1 regulated ELF4 expression by competitively binds to miR-365. PVT1 regulated the stemness and sensitivity of TMZ of glioma cells through miR-365/ELF4/ SOX2 axis. This study identified that PVT1 promoted glioma stemness through miR-365/ELF4/SOX2 axis.

Measurement of femoral neck anteversion of developmental dislocation of hip in children by 3D printing technique / 中国骨伤

OBJECTIVE@#To explore the method and accuracy of measuring the femoral neck anteversion in children with developmental dislocation of the hip by using 3D printing technology, so as to find out the method of measuring the femoral neck anteversion accurately and guide the formulation of the operation plan.@*METHODS@#From June 2016 to September 2018, 17 patients with unilateral developmental dislocation of the hip were enrolled in the study, including 2 males and 15 females, aged 2 to 13 (5.47±0.81) years old, 11 on the left and 6 on the right. The methods of CT, 3D printing and intraoperative anteversion of femoral neck were used to measure the anteversion of femoral neck respectively. The intraoperative measurement was used as the standard reference value forstatistical analysis, and the accuracy of the first two methods was compared.@*RESULTS@#The average value of CT was (36.00±1.66)°, the average value of model method was(43.91±1.62)°, and the average value of intraoperative method was(44.21±1.62)°. There were significant differences in CT measurement, model measurement and intraoperative measurement(0.05).@*CONCLUSION@#Compared with the traditional CT method, the 3D printing model method is simpler, more accurate and more repeatable, and can simulate the operation before operation. It is an ideal method to measure the femoral neck anteversion.

Influencing factors for depressive symptoms in the elderly aged 65 years and older in 8 longevity areas in China / 中华流行病学杂志

To analyze influencing factors for depressive symptoms in the elderly aged 65 years and older in 8 longevity areas in China. We recruited 2 180 participants aged 65 years and older in 8 longevity areas from Healthy Aging and Biomarkers Cohort Study, a sub-cohort of the Chinese Longitudinal Healthy Longevity Survey in 2017. Multivariate logistic regression analysis was performed to evaluate the relationships of socio-demographic characteristics, behavioral lifestyle, chronic disease prevalence, functional status, family and social support with depressive symptoms in the elderly. The detection rate of depression symptoms was 15.0 in the elderly aged 65 years and older in 8 longevity areas of China, and the detection rate of depression symptoms was 11.5 in men and 18.5 in women. Multivariate logistic regression analysis results showed that the detection rate of depressive symptoms was lower in the elderly who had regular physical exercises (=0.44, 95: 0.26-0.74), frequent fish intakes (=0.57, 95: 0.39-0.83), recreational activities (=0.65, 95: 0.44-0.96), social activities (=0.28, 95: 0.11-0.73) and community services (=0.68, 95: 0.50-0.93). The elderly who were lack of sleep (=2.04, 95: 1.49-2.80), had visual impairment (=1.54, 95: 1.08-2.18), had gastrointestinal ulcer (=2.97, 95: 1.53-5.77), had arthritis (=2.63, 95: 1.61-4.32), had higher family expenditure than income (=1.80, 95: 1.17-2.78) and were in poor economic condition (=4.58, 95: 2.48-8.47) had higher detection rate of depressive symptoms. The status of doing physical exercise, fish intake in diet, social activity participation, sleep quality or vision, and the prevalence of gastrointestinal ulcers and arthritis were associated with the detection rate of depressive symptoms in the elderly.

Current status of body mass index and related influencing factors in the elderly aged 65 years and older in 8 longevity areas in China / 中华流行病学杂志

To understand the current status of BMI of the elderly and related factors in longevity areas in China, and provide scientific evidence for the control of BMI level in elderly population. Data used in this study were obtained from Healthy Aging and Biomarkers Cohort Study, a sub-cohort of the Chinese Longitudinal Healthy Longevity Survey. A total of 2 825 elderly in 8 longevity areas in China were surveyed and measured in 2017. The BMI levels of 2 217 elderly aged 65 years and older were calculated and in follow up. The ordered classification logistic regression model was used to analyze the influencd factors for the BMI in the elderly. The BMI of the elderly in 8 longevity areas in China was (22.36±3.87) kg/m(2), and it was (22.76±3.58) kg/m(2) for males and (21.75±3.98) kg/m(2) for females. The BMI levels were normal in 1 165 elderly persons. The prevalence of underweight, overweight and obesity were 15.8, 24.0 and 7.7, respectively. Multivariate analysis showed that the main factors affecting the BMI of people under 100- years old were age (65-: =2.78, 95: 1.87-4.15; 80-: =1.47, 95: 1.00-2.17), smoking status (=0.46, 95: 0.32-0.66), annual household income (<30 000 Yuan: =1.26, 95: 1.07-1.47; 30 000-70 000 Yuan: =1.52, 95: 1.12-1.86), and frequency of tea intake(=1.36, 95: 1.01-1.71), while the factor in people aged ≥100 years was gender (=3.68, 95: 1.32-10.36). The prevalence of underweight, overweight and obesity were high in the elderly from longevity areas in China. It is necessary to pay attention to the trend of overweight and obesity due to smoking, higher annual household income and regular tea drinking in the elderly men.

Influence of visual impairment on mortality in the elderly aged 65 years and older in 8 longevity areas in China / 中华流行病学杂志

To understand the relationship between visual impairment and risk of all-cause mortality in the elderly aged 65 years and older in 8 longevity areas in China. The data of the elderly aged 65 years and older in the project in 2012 were obtained from Healthy Aging and Biomarkers Cohort Study, a sub-cohort of the Chinese Longitudinal Healthy Longevity Survey, including physical measurement and survival status, and a follow-up for survival outcomes were conducted in 2014 and 2017 respectively. Cox proportional hazard regression model was used to analyze the influence of visual impairment on mortality. Gender and age specific analysis was conducted. A total of 1 736 elderly adults were included. A total of 943 deaths occurred during the 5-year follow-up period with a 5-year mortality rate of 54.3. The 5-year mortality rate was 76.7 in the group with visual impairment, and 47.6 in the group without visual impairment (<0.001). After adjusting for demographic information, life style and some disease factors, the risk of 5-year mortality in the group with visual impairment group was 1.30 times higher than that in the group without visual impairment (=1.30, 95: 1.09-1.55). In the females, the risk for mortality in the group with visual impairment was 1.48 times higher than that in the group without visual impairment (=1.48, 95:1.20-1.84). However, vision status was not associated with the risk for mortality in males (=1.02, 95: 0.72-1.43). The risk for mortality in the group with visual impairment was 1.39 times higher than that in the group without visual impairment in the elderly aged over 90 years (=1.39, 95: 1.13-1.70). Vision status was not associated with mortality risk in the elderly aged 65-79 years and 80-89 years (=1.37, 95: 0.61-3.07; =0.95, 95: 0.61-1.48). In the elderly people in China, visual impairment is a risk factor for mortality.

Effects of estimated glomerular filtration rate on all-cause mortality in the elderly aged 65 years and older in 8 longevity areas in China / 中华流行病学杂志

To investigate the association between estimated glomerular filtration rate (eGFR) and all-cause mortality in the elderly aged 65 years and older in longevity areas in China. Data used in this study were obtained from Healthy Aging and Biomarkers Cohort Study, a sub-cohort of the Chinese Longitudinal Healthy Longevity Survey, 1 802 elderly adults were collected in the study during 2012-2017/2018. In this study, the elderly were classified into 4 groups, moderate-to-severe group [<45 ml·min(-1)·(1.73 m(2))(-1)], mild-to-moderate group [45- ml·min(-1)·(1.73 m(2))(-1)], mild group [60- ml·min(-1)·(1.73 m(2))(-1)] and normal group [≥90 ml·min(-1)·(1.73 m(2))(-1)] according to their eGFR levels. After 6 years of follow-up, 852 participants died, with a mortality rate of 47.3. Multivariate Cox regression analysis showed that the levels of eGFR were negatively correlated with all-cause mortality risk in the elderly (the of elderly was 0.993 and the 95 was 0.989-0.997 for every unit of eGFR increased, =0.001), while compared with the group with normal eGFR, the (95) of the elderly in the moderate-to-severe group, mild-to-moderate group, and mild group were 1.690 (1.224-2.332, =0.001), 1.312 (0.978-1.758, =0.070), 1.349 (1.047-1.737, =0.020) respectively [trend test <0.001]. The decrease in eGFR was associated with higher mortality risk among the elderly in longevity areas in China.

Prediction of 6-year incidence risk of chronic kidney disease in the elderly aged 65 years and older in 8 longevity areas in China / 中华流行病学杂志

To establish a prediction model for 6-year incidence risk of chronic kidney disease (CKD) in the elderly aged 65 years and older in China. In this prospective cohort study, we used the data of 3 742 participants collected during 2008/2009-2014 and during 2012-2017/2018 from Healthy Aging and Biomarkers Cohort Study, a sub-cohort of the Chinese Longitudinal Healthy Longevity Survey. Two follow up surveys for renal function were successfully conducted for 1 055 participants without CKD in baseline survey. Lasso method was used for the selection of risk factors. The risk prediction model of CKD was established by using Cox proportional hazards regression models and visualized through nomogram tool. Bootstrap method (1 000 resample) was used for internal validation, and the performance of the model was assessed by C-index and calibration curve. The mean age of participants was (80.8±11.4) years. In 4 797 person years of follow up, CKD was found in 262 participants (24.8). Age, BMI, sex, education level, marital status, having retirement pension or insurance, hypertension prevalence, blood uric acid, blood urea nitrogen and total cholesterol levels and estimated glomerular filtration rate in baseline survey were used in the model to predict the 6-year incidence risk of CKD in the elderly. The corrected C-index was 0.766, the calibration curve showed good consistence between predicted probability and observed probability in high risk group, but relatively poor consistence in low risk group. The incidence risk prediction model of CKD established in this study has a good performance, and the nomogram can be used as visualization tool to predict the 6-year risk of CKD in the elderly aged 65 years and older in China.

Expression and purification of soluble recombinant antibody fragment and immunotoxin in Escherichia coli using Npu DnaE C-cleavage mutant / 中国药科大学学报

@#The purpose of this experiment is to achieve the soluble expression of immunotoxins in Escherichia coli and avoid the complicated operation caused by the formation of inclusion bodies. An anti-mesothelin monoclonal antibody SS1 and it′s derivative SS1P which composing of SS1 and a truncated pseudomonas exotoxin PE38KDEL were used as the passenger protein. We took advantages of solubility promoting fusion tags and the self-cleaving split intein in recombinant antibody fragment and immunotoxin expression and purification. We constructed solubilizing tags-NpuCD118G fusion tags, and recombinant SS1/SS1P were fused at the C-terminal of the fusion tags. The constructs were expressed in E. coli(SHuffle T7)cytoplasm in soluble form at low temperature. Dextrin Beads 6FF column and Nickel column was used to purify the fusion protein. The self-cleavage of the fusion protein was achieved by adding the NpuN fragment. The released solubilizing tag and unreacted precursor were removed by Nickel column and the cleaved antibody fragment and immunotoxinwere further captured by Capto L. The dissociation constant of the obtained Fv and immunotoxin were determined by Fortebio. In summary, the current method could enhance the solubility of antibody fragment and immunotoxin in E. coli, and could improve the purification process. This method provides a reference for the development of a method for soluble expression and purification of immunotoxin-type pharmaceutical recombinant proteins based on the Escherichia coli expression system.

Improving proteolytic stability of Npu DnaE C-fragment in Escherichia coli expression system / 中国药科大学学报

@#Naturally split Npu DnaE intein can mediate rapid trans-splicing and C-cleavage, which is of great use in many aspects of protein engineering. However, the degradation of NpuC during expression and purification reduces the yield and purity of recombinant protein. N2C, an extended NpuN2-containing N-terminal NpuC fragment, was constructed to improve NpuC stability. N2C was expressed in BL21(DE3) and purified by affinity chromatography. The degradation ratio was calculated by ImageJ, and the factors affecting the C-terminal cleavage reaction of intein, such as temperature, DTT concentration and N/C ratio, were also investigated. The results showed that N2C lowered the proportion of degradation to 2.7%-7.2% and the yield of C-terminal cleavage reached 90% in 30 min at 37 °C with an N/C ratio of 5∶1 catalyzed by 1 mmol/L DTT. N2C can not only improve the stability of NpuC in Escherichia coli expression system, but also retain the activity of C-terminal cleavage reaction, which is of great significance for its application in protein purification.

Splicing reaction of split inteins between non-natural exteins containing amino acid sequences of antibody hinge area / 中国药科大学学报

@#Intein is a functional protein that mediates self-cleavage from precursor protein and simultaneously connects the exteins on both sides of the intein via peptide bonds. Among all kinds of inteins, split intein has a wide range of applications in the field of antibody ligation. However, since the naturally split intein specifically recognizes the first three amino acid sequences “cysteine, phenylalanine, and asparagine”(CFN)of the extein, exogenous amino acids are inevitably introduced after the protein splicing reaction. In this study, the amino acid sequence “cysteine, aspartic acid, and lysine”(CDK)of the antibody hinge region was substituted for “CFN” as the recognition site for the intein and the split intein Npu DnaE was mutated into Npu*GEP DnaE. The results showed that the mutant could recognize “CDK” and the intein splicing reaction could successfully take place. The factors affecting the intein splicing reaction platform, such as pH, temperature and the concentration of NaCl and DTT were investigated in this study. The results showed that the splicing reaction of the mutant performed well, which indicated its potential usefulness in bispecific antibody assembly. In conclusion, the problem of introducing foreign amino acids was alleviated, the broadness of intein substrate was further expanded, and further technical support for the application of intein to the antibody assembly was provided.

Analysis on the delayed effect of candesartan ester combined with tripterygium wilfordii on chronic renal failure / 中国基层医药

Objective@#To analyze the delayed effect of candesartan ester combined with tripterygium wilfordii on renal failure of chronic kidney disease (CKD) from 2 to 3.@*Methods@#From May 2015 to May 2016, 80 patients with chronic renal insufficiency (CRF) in Jiaxing Hospital of Zhejiang Provincial Armed Police Corps were selected in the research.The patients were randomly divided into the combined group and control group by compute random grouping method, with 40 cases in each group.The combined group was given candesartan ester combined with tripterygium wilfordii glycosides, and the control group was given glucocorticoid treatment.The therapeutic effect, the incidence of adverse reactions, improvement of clinical indicators and degree of kidney damage were compared between the two groups.@*Results@#The total effective rate of the combined group was 95%, which was higher than 80% of the control group (χ2=4.116, P<0.05). The incidence of adverse reactions in the combined group was 7.5%, which was significantly lower than that in the control group (χ2=6.657, P<0.05). The biochemical parameters of the two groups had no statistically significant differences(t=1.032, 0.835, 1.017, 0.924, all P>0.05), but the serum urea nitrogen (BUN), serum creatinine (Scr), glomerular filtration rate (GFR) and uric acid levels in the combined group were (6.3±1.2)mmol/L, (99.6±4.5)μmol/L, (82.7±6.5)mL/min and (335.2±28.6)μmol/L, respectively, which were significantly better than those of the control group[(7.9±2.5)mmol/L, (128.9±6.4)moL/L, (59.6±7.3)mL/min and (437.9±37.1)mol/L](t=7.812, 8.119, 7.523, 8.981, all P<0.05). Before treatment, the renal damage degree between the two groups had no statistically significant difference (P>0.05). After treatment, the proportions of renal damage degree stage Ⅰ, Ⅱ, Ⅲ and Ⅳ in the combined group were 72.5%, 12.5%, 15.0% and 0.00%, respectively, which were significantly lower than those in the control group (22.5%, 32.5%, 37.5% and 7.5%) (χ2=20.058, 4.592, 5.237, 1.394, all P<0.05).@*Conclusion@#Candesartan ester combined with tripterygium wilfordii can be effectively used in the treatment of CKD at 2 to 3 stages, and the incidence of adverse reactions of drugs is relatively low, which can effectively improve the clinical indicators of patients and has a good retarding effect on renal failure.Therefore, it is recommended to be widely used in clinical treatment.