Objective:This study aimed to explore the feasibility and clinical value of monitoring the progression of early kidney injury in type 2 diabetic patients by assessment of the urinary C-terminal agrin fragment (uCAF) with enzymatic chemiluminescence immunoassay.Methods:A total of 251 patients with type 2 diabetes, who attended the Second Affiliated Hospital of Wenzhou Medical University from October 2018 to March 2020, were included in this retrospective analysis. One hundred and fifty-six participants undergoing health check-up at the Second Affiliated Hospital of Zhejiang University School of Medicine in February 2021 served as controls. Basic clinical information, glycosylated hemoglobin type A 1c and serum creatinine values were recorded, and urine specimens were collected for urinary creatinine, urinary α 1 microglobulin(uα 1M), urinary immunoglobulin G (uIgG), urinary albumin, urinary N-Acetyl-B-D-glycosaminidase (uNAG) and uCAF measurements. Based on the estimated glomerular filtration rate (eGFR), 251 patients were classified into G1~G5 stage groups with 116, 22, 28, 55 and 30 patients in each group. One hundred and sixty-six patients with early diabetic kidney disease (stage G1-G3) were divided into subgroups A1 (79), A2 (48) and A3 (39) according to the urinary albumin/creatinine ratio (UACR), the uα1M levels were divided into uα1M subgroup 1 (83 cases), uα1M subgroup 2 (42 cases), and uα1M subgroup 3 (41 cases), and uIgG subgroup 1 (83 cases), uIgG subgroup 2 (42 cases), and uIgG subgroup 3 (41 cases) according to uIgG levels. The Spearman method was used to analyze the correlation between uCAF levels and eGFR, UACR, uα1M and uIgG levels. Results:(1) The linear range of the uCAF detected by enzymatic chemiluminescence immunoassay was 3.97-2 000.00 ng/ml, with a detection limit of 2.28 ng/ml, intra-batch coefficients of variation of 1.15% and 1.57%, inter-batch coefficients of variation of 1.63% and 5.78%, and a biological reference interval of <95.35 μg/g Cr. (2) The uCAF level and positive rate (UACR≥30 mg/g) increased with the decrease of eGFR from G1-G3, uCAF level was negatively correlated with eGFR value ( r=-0.543, P<0.000 1), and the positive rate increased from 24.14% (28/116) to 85.71% (24/28) from G1-G3. The uCAF level and positivity rate decreased with the decrease of eGFR from G4 to G5. uCAF level was positively correlated with eGFR value ( r=0.495, P<0.001), and the positivity rate decreased from 30.91% (17/55) to 23.33% (7/30) from G4 to G5. (3) In patients with early diabetic kidney disease, uCAF levels and positivity rates increased gradually with the increase of UACR. uCAF levels were positively correlated with UACR values ( r=0.602, P<0.001), and the uCAF positivity rate reached 21.52% (17/79) in the A1 subgroup. (4) uCAF level was positively correlated with uα1M and uIgG levels in patients with early diabetic kidney disease ( r=0.757, 0.596, both P<0.001). Conclusion:Analytical performance of enzyme chemiluminescence immunoassay for the detection of CAF is satisfactory and could be used a biomarker for monitoring damage and progression of early diabetic kidney disease in patients with type 2 diabetes.

ObjectiveTo investigate the effect of Liuwei Dihuangwan on memory function of senescence-accelerated mouse prone 8 （SAMP8） mice by regulating autophagy through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/forkhead box O3a （FoxO3a） pathway. MethodSix male senescence-accelerated mouse resistant 1 （SAMR1） mice of SPF grade aging 6 months were assigned to a normal group， and 24 male SAMP8 mice of SPF grade aging 6 months were randomly divided into a model group， a donepezil group （0.747 mg·kg^-1）， and high- and low-dose Liuwei Dihuangwan groups （2.700 and 1.350 g·kg^-1）， with 6 mice in each group. The mice were treated with drugs by gavage for 2 months. Morris water maze was used to detect the learning and memory abilities of mice in each group. Nissl staining was used to observe the neurons in the cortex and hippocampus. The positive expression of microtubule-associated protein 1 light chain 3B （LC3B） in the cortex and hippocampus was detected by immunohistochemistry （IHC）. Western blot was used to detect the protein expression of the mammalian ortholog of yeast ATG6 （Beclin-1）， B cell lymphoma-2 （Bcl-2）， autophagy-related gene 5 （ATG5）， cysteinyl aspartate-specific protease 3 （Caspase-3）， Caspase-9， Akt， p-Akt， FoxO3a， and p-FoxO3a. ResultCompared with the normal group， the model group showed prolonged escape latency （P<0.05，P<0.01）， reduced number of platform crossings and the residence time in the target quadrant （P<0.01）， decreased neurons with reduced volume and dispersed distribution in the cortex and hippocampus， increased positive expression of LC3B （P<0.01）， elevated expression of Beclin-1 and ATG5 in the cortex （P<0.01）， declined Bcl-2 expression （P<0.01）， up-regulated Caspase-3 and Caspase-9 expression （P<0.01）， and decreased expression levels of p-Akt/Akt and p-FoxO3a/FoxO3a （P<0.01）. Compared with the model group， the donepezil group and the Liuwei Dihuangwan groups showed shortened 3 d escape latency （P<0.05，P<0.01）， increased number of platform crossings （P<0.01）， and prolonged residence time in the target quadrant （P<0.01）. In the donepezil group， the number of neurons in the cortex and hippocampus was increased. In the Liuwei Dihuangwan groups， the number of neurons and Nissl bodies increased with denser distribution and lower degree of cell damage. The positive expression of LC3B in the cortex and hippocampus was decreased in the donepezil group and Liuwei Dihuangwan groups （P<0.01）. The expression of Beclin-1 was decreased in the Liuwei Dihuangwan groups （P<0.01）. The expression of ATG5 was decreased in the donepezil group and the low-dose Liuwei Dihuangwan group （P<0.01）. The donepezil group and the Liuwei Dihuangwan groups showed the increased expression level of Bcl-2 in the cortex （P<0.01）， decreased expression level of Caspase-3 （P<0.01）， reduced expression level of Caspase-9 （P<0.05，P<0.01）， and elevated expression levels of p-Akt/Akt and p-FoxO3a/FoxO3a （P<0.01）. ConclusionLiuwei Dihuangwan can effectively improve the learning and memory abilities of the SAMP8 mice and protect neurons. Its mechanism may be related to the regulation of the PI3K/Akt/FoxO3a signaling pathway， down-regulation of the expression of ATG5， Beclin-1， and LC3B， and the inhibition of apoptosis.

ObjectiveTo investigate the protective effect of Liuwei Dihuangwan on neurovascular injury in SAMP8 mice. MethodThe Alzheimer's disease （AD） model with insufficiency of kidney essence was induced in 75 SAMP8 mice aging 6 months. The model mice were divided into model group， positive control group （donepezil hydrochloride， 0.747 mg·kg^-1·d^-1）， and high-， medium-， and low-dose Liuwei Dihuangwan groups （2.700， 1.350， 0.675 g·kg^-1·d^-1）， with 15 mice in each group. Fifteen SAMR1 mice were assigned to a normal control group. All mice were administered continuously for 2 months. The spatial memory of mice was tested by the Morris water maze. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in the hippocampus and cortex of brain tissues. The immunohistochemical method （IHC） was used to detect the deposition of amyloid β-protein （Aβ） and the expression of von Willebrand factor （vWF） and CD34 in the hippocampus and cortex of brain tissues. Electron microscopy was used to observe the ultrastructural changes in cerebral microvessels. Western blot was used to detect the protein expression levels of the receptor of advanced glycation endproduct （RAGE）， low-density lipoprotein receptor-related protein 1 （LRP1）， vascular endothelial growth factor A （VEGF-A）， and P-selection in the hippocampus and cortex of brain tissues. ResultCompared with the normal control group， the model group showed prolonged escape latency and swimming distance （P<0.01）， increased number of glial cells， decreased number of nerve cells， blurred tight junctions or enlarged gap of the brain microvascular endothelial cells， severely injured membrane structure， swollen mitochondria of endothelial cells， ruptured membrane， massive dissolution in cristae， increased protein expression of Aβ and vWF in the hippocampus and cortex （P<0.01）， reduced protein expression of CD34 （P<0.05）， elevated protein expression of RAGE and P-selection in the cortex （P<0.01）， and decreased protein expression level of LRP1 and VEGF-A （P<0.01）. Compared with the model group， the Liuwei Dihuangwan groups showed shortened escape latency and swimming distance （P<0.05）， reduced number of glial cells in the cortex and hippocampus， increased number of microvessels in the cortex， clear double-layer membrane structure in tight junctions between the microvascular endothelial cells， increased number of mitochondria with intact membrane and recovered mitochondrial cristae， decreased protein expression of Aβ， vWF， RAGE， and P-selection in the hippocampus and cortex （P<0.05）， and increased protein expression of CD34， LRP1， and VEGF-A （P<0.05）. ConclusionLiuwei Dihuangwan can regulate Aβ metabolism through the RAGE/LRP1 receptor system and promote cerebral microvascular angiogenesis by inhibiting vWF expression and increasing VEGF-A and CD34， thereby improving cerebral microvascular injury in SAMP8 mice.

This study aimed to quantitatively assess the effectiveness of the Wuhan lockdown measure on controlling the spread of coronavirus diesase 2019 (COVID-19). : Firstly，estimate the daily new infection rate in Wuhan before January 23，2020 when the city went into lockdown by consulting the data of Wuhan population mobility and the number of cases imported from Wuhan in 217 cities of Mainland China. Then estimate what the daily new infection rate would have been in Wuhan from January 24 to January 30th if the lockdown measure had been delayed for 7 days，assuming that the daily new infection in Wuhan after January 23 increased in a high，moderate and low trend respectively (using exponential, linear and logarithm growth models). Based on that，calculate the number of infection cases imported from Wuhan during this period. Finally，predict the possible impact of 7-day delayed lockdown in Wuhan on the epidemic situation in China using the susceptible-exposed-infectious-removed (SEIR) model. : The daily new infection rate in Wuhan was estimated to be 0.021%，0.026%，0.029%，0.033% and 0.070% respectively from January 19 to January 23. And there were at least 20 066 infection cases in Wuhan by January 23，2020. If Wuhan lockdown measure had been delayed for 7 days，the daily new infection rate on January 30 would have been 0.335% in the exponential growth model，0.129% in the linear growth model，and 0.070% in the logarithm growth model. Correspondingly，there would have been 32 075，24 819 and 20 334 infection cases travelling from Wuhan to other areas of Mainland China，and the number of cumulative confirmed cases as of March 19 in Mainland China would have been 3.3-3.9 times of the officially reported number. Conclusions: Timely taking city-level lockdown measure in Wuhan in the early stage of COVID-19 outbreak is essential in containing the spread of the disease in China.
COVID-19 ; China/epidemiology* ; Cities ; Communicable Disease Control ; Humans ; SARS-CoV-2

To predict the epidemiological trend of coronavirus disease 2019 (COVID-19) by mathematical modeling based on the population mobility and the epidemic prevention and control measures. : As of February 8，2020，the information of 151 confirmed cases in Yueqing，Zhejiang province were obtained，including patients' infection process，population mobility between Yueqing and Wuhan，etc. To simulate and predict the development trend of COVID-19 in Yueqing, the study established two-stage mathematical models，integrating the population mobility data with the date of symptom appearance of confirmed cases and the transmission dynamics of imported and local cases. : It was found that in the early stage of the pandemic，the number of daily imported cases from Wuhan (using the date of symptom appearance) was positively associated with the number of population travelling from Wuhan to Yueqing on the same day and 6 and 9 days before that. The study predicted that the final outbreak size in Yueqing would be 170 according to the number of imported cases estimated by consulting the population number travelling from Wuhan to Yueqing and the susceptible-exposed-infectious-recovered (SEIR) model; while the number would be 165 if using the reported daily number of imported cases. These estimates were close to the 170，the actual monitoring number of cases in Yueqing as of April 27，2020. : The two-stage modeling approach used in this study can accurately predict COVID-19 epidemiological trend.
COVID-19 ; China/epidemiology* ; Disease Outbreaks ; Humans ; Models, Theoretical ; Pandemics ; SARS-CoV-2

Objective:To report the diagnosis and surgical treatment of a special type of Maisonneuve injury.Methods:A total of 4 patients were treated at Department of Orthopaedics, Beijing Chao Yang Hospital from January 2015 to July 2019 for Maisonneuve injury. They were 3 males and one female, aged from 34 to 61 years (average, 45.3 years). All injuries were closed, initially manifested as posterior dislocation of the ankle on X-ray films and X-ray and CT re-exams after manual reduction showed fine reduction with no obvious fracture of the ankle joint. Consequently the diagnosis of their Maisonneuve injury was missed in emergency visits, but re-exams in outpatient visits showed separation of distal tibiofibular syndesmosis. All the patients were treated by restoration of the fibular length, fixation of the distal tibiofibular syndesmosis and repair of the triangular ligament. American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score was used to evaluate functional recovery of the ankle joint at 12 months after operation.Results:All the 4 patients were followed up for 12 to 14 months (average, 12.7 months). The fractures united after 110 to 185 days (average, 149.3 days). No post-operative complications like infection, delayed union or nonunion were observed. The AOFAS score at 12 months ranged from 82 to 96 points (average, 90.5 points), giving 3 excellent and one good cases.Conclusions:The Maisonneuve injury which is clinically manifested as posterior dislocation of the ankle with no obvious fracture of the ankle joint is likely to be missed in clinical diagnosis. Therefore, X-ray exam of the full length tibia and fibula should be taken in physical examination after reduction for the patients with simple posterior dislocation of the ankle. Once the special type of Maisonneuve injury is diagnosed, surgical treatment is indicated. Satisfactory treatment efficacy can be achieved by fixation of the distal tibiofibular syndesmosis and repair of the triangular ligament.

Objective: To analyze the influencing factors for hospitalization of pneumoconiosis patients, so as to provide reference for improving the timeliness of treatment and quality of life in these patients. Methods: The pneumoconiosis patients in Changshan, Wenling and Changxing ( having mining industry ) of Zhejiang Province were selected by stratified and typical sampling methods. Their demographic features, prevalence of pneumoconiosis and hospitalization status were collected by a questionnaire survey. Logistic regression analysis was conducted to analyze the influencing factors for hospitalization. Results: Among 166 pneumoconiosis patients, 122 ( 73.49% ) chose to be hospitalized. The multivariate logistic regression analysis revealed that the patients who scored higher in ED-5Q ( OR=65.821, 95%CI: 3.831-1130.781 )and who would communicate with other pneumoconiosis patients ( OR=3.744, 95%CI: 1.261-11.115 ) were more likely to be hospitalized; the patients who were under the medical insurance for urban employees were more likely to be hospitalized than those who were under the new rural cooperative medical care system ( OR=0.033, 95%CI: 0.007-0.154 ) and others ( OR=0.038, 95%CI: 0.002-0.960 ) . Conclusion The quality of life, communication with other pneumoconiosis patients and type of medical insurance were the influencing factors for hospitalization of pneumoconiosis patients.

Objective:To explore the occupational stress status and influencing factors of workers in a textile factory in Zhejiang Provice.Methods:In October 2018, 505 workers from a textile factory in Zhejiang Province were selected as research objects by convenient sampling method. A total of 505 questionnaires were distributed, 495 of which were effective and the effective recovery rate was 98.0%. Job Content Questionnaire (JCQ) and Effort Reward Imbalance Questionnaire (REI) were used to investigate textile workers' occupational stress and analyze its influencing factors. The differences of the composition ratio of different groups were tested by χ 2 test. The influencing factors such as age, gender and occupation on occupational stress were analyzed by multivariate logistic regression. Results:ERI analysis results showed that the high and low occupational stress accounted for 30.1% and 69.9%, respectively. The differences of occupational stress among workers of different job types and working ages were statistically significant ( P<0.05) . The detection rates of high occupational stress of paper workers and spinners were 47.8% (11/23) and 44.8% (30/67) , respectively, higher than other jobs. The detection rate of high occupational stress for workers with more than 5 years of service was 46.4% (13/28) . The results of JCQ analysis showed that there was no statistical significance in the differences of daily working hours and length of service between different gender, education levels, types of work patterns, and occupational stress ( P>0.05) . Job types had significant effects on the occupational stress defined by ERI ( P<0.05) , the risk of occupational stress was 2.151 times than that of the coiler. Conclusion:There are significant differences in occupational stress risk among workers of different types of work in textile industry, so different measures should be taken to prevent and control occupational stress in different jobs.

Objective:To explore the occupational stress status and influencing factors of workers in a textile factory in Zhejiang Provice.Methods:In October 2018, 505 workers from a textile factory in Zhejiang Province were selected as research objects by convenient sampling method. A total of 505 questionnaires were distributed, 495 of which were effective and the effective recovery rate was 98.0%. Job Content Questionnaire (JCQ) and Effort Reward Imbalance Questionnaire (REI) were used to investigate textile workers' occupational stress and analyze its influencing factors. The differences of the composition ratio of different groups were tested by χ 2 test. The influencing factors such as age, gender and occupation on occupational stress were analyzed by multivariate logistic regression. Results:ERI analysis results showed that the high and low occupational stress accounted for 30.1% and 69.9%, respectively. The differences of occupational stress among workers of different job types and working ages were statistically significant ( P<0.05) . The detection rates of high occupational stress of paper workers and spinners were 47.8% (11/23) and 44.8% (30/67) , respectively, higher than other jobs. The detection rate of high occupational stress for workers with more than 5 years of service was 46.4% (13/28) . The results of JCQ analysis showed that there was no statistical significance in the differences of daily working hours and length of service between different gender, education levels, types of work patterns, and occupational stress ( P>0.05) . Job types had significant effects on the occupational stress defined by ERI ( P<0.05) , the risk of occupational stress was 2.151 times than that of the coiler. Conclusion:There are significant differences in occupational stress risk among workers of different types of work in textile industry, so different measures should be taken to prevent and control occupational stress in different jobs.

Objective: To establish a microwave scattering parameter acquisition system to detect cerebral hemorrhage and cerebral ischemia animal models, and to study the non-contact rapid identification methods for the two stroke types. Methods: Rabbits were selected for modeling. Eight rabbits in the cerebral hemorrhage group were injected with autologous blood. Six rabbits in the cerebral ischemia group were treated with bilateral common carotid artery clamping and femoral artery bleeding. The measurement excitation source has a scanning frequency range of 300 kHz to 3 GHz and an intermediate frequency bandwidth of 30 kHz. The signal of the S21 phase was acquired. The collected microwave scattering signals were subjected to mean filtering, principal component analysis dimension reduction, and mean clustering and nearest neighbor analysis to realize the identification of stroke types. Results: The microwave scattering measurement method can reflect the changes of cerebral hemorrhage and cerebral ischemia. The phase of S21 decreases with the increase of blood loss and increases with the increase of ischemic duration. The results of the differential experiment showed that all 14 models were correctly identified. Conclusions The stroke identification system based on microwave scattering measurement can effectively distinguish rabbit cerebral hemorrhage model and ischemic model. This technology is low cost, portable non-invasive, simple operation and fast, which make it be a promising method for identifying pre-hospital stroke types.