【Objective】 To explore biosafety countermeasures and optimize the sample detection procedures for the COVID-19 in tertiary hospitals in Wuhan, so as to provide effective protection for blood transfusion laboratory. 【Methods】 Referring to General Requirements for Laboratory Biosafety, Novel Coronavirus Pneumonia Prevention and Control Program (Fourth Edition), Technical Guide for Laboratory Testing of the Novel Coronavirus Pneumonia (Fourth Edition), the Novel Coronavirus Laboratory Biology Safety Guidelines (Second Edition) and other technical guidelines and expert consensus, relevant biosafety protection measures and testing procedures were formulated in combination of the layout and conditions of our laboratory. 【Results】 The biosafety zoning of blood transfusion laboratory, procedures for sample receiving, testing, and sterilizing, as well as the protection procedures for workers entering and exiting the laboratory were formulated, with good adherence with the relevant laws, regulations and technical specifications concerning COVID-19. During the prevention and treatment of the COVID-19, the work of blood transfusion department was carried out orderly, without any medical staff infection. 【Conclusion】 The biosafety protection measures and sample detection procedures in compliance with relevant laws, regulations and technical specifications, taking the actual condition of our laboratory into consideration, can effectively prevent and control the infection of COVID-19 and protect the health of medical staff.

Objective: To provide data reference for the clinical diagnosis of COVID-19, promote the innovation and improvement of antibody detection technology of COVID-19, and discuss the application of antibody detection of new coronavirus by evaluating the sensitivity, specificity, compliance rate and limit of antibody detection in 10 SARS-CoV-2 antibody detection kits. Methods: According to the instructions of 10 new coronavirus antibody detection kits, the specific IgM and IgG in 74 serum samples were detect- ed, and the limit of detection of each kit was tested with 10 samples diluted by a 2-fold gradient. Results: The coincidence rate of 10 kits(product A-product J)were 89.2%(66/74), 89.2%(66/74), 86.5%(64/74), 95.9%(71/74), 52.7%(39/74), 75.7%(56/74), 86.5%(64/74), 79.7%(59/74), 50.0%(37/74), 20.3%(15/74), respectively. In the detection limit tests for the kits A-J with the 10 of 2x gradient diluted samples, the performance of kit C was the best, with the limit of detection for IgM 0.16AU/ml and the limit of detection for IgG 1.00 AU/ml. Conclusion: There are significant differences in sensitivity, specificity, compliance and limit of detection of 10 new coronavirus antibody testing reagents, and the product of high sensitivity, specificity, and compliance rates should be selected for clinical applications to ensure the accuracy of test results.

Objective: To explore the rats bone biomechanics improvement action after exercise combined with traditional Chinese medicine compound of castration. Method: A total of 100 SPF SD female rats, aged 180 days, were randomly divided into blank group，model group，exercise，traditional Chinese medicine compound，sports combined with traditional Chinese medicine compound group, 20 in each group. Followed conventional breeding 14 days after surgery. 2ml 0.9% sodium chloride was filled to stomach daily for model group and exercise group. 2ml traditional Chinese medicine compound solution was filled to the stomach daily for the traditional Chinese medicine compound and sports combined with traditional Chinese medicine compound group. Besides, the rats were forced to running on flat treadmill 20 minutes daily at speed of 5m/min for exercise group and sports combined with traditional Chinese medicine compound group. After 12 weeks treatment, bilateral femur were collected, attached muscle and connective tissue were took out. Then three-point bending test, microstructure observation, and static parameters of bone trabecula test were completed. Result: BMD, maximal load of femur, and static parameters of bone trabecula test for model group, blank group, exercise group, Chinese medicine compound group, exercise combined with traditional Chinese medicine compound group all have obvious difference from the model group (P<0.01 or P<0.05). Conclusion: Exercise combined with traditional Chinese medicine compound is beneficial for treatment of osteoporosis in castrated rats.

BACKGROUND: Drug resistance is the main cause of high mortality of lung cancer. This study was conducted to investigate the effect of folic acid (FA) on the resistance of non-small cell lung cancer (NSCLC) cells to Osimertinib (OSM) by regulating the methylation of dual specificity phosphatase 1 (DUSP1). METHODS: The OSM resistant NSCLC cell line PC9R was establishd by gradually escalation of OSM concentration in PC9 cells. PC9R cells were randomly grouped into Control group, OSM group (5 μmol/L OSM), FA group (600 nmol/L FA), methylation inhibitor decitabine (DAC) group (10 μmol/L DAC), FA+OSM group (600 nmol/L FA+5 μmol/L OSM), and FA+OSM+DAC group (600 nmol/L FA+5 μmol/L OSM+10 μmol/L DAC). CCK-8 method was applied to detect cell proliferation ability. Scratch test was applied to test the ability of cell migration. Transwell assay was applied to detect cell invasion ability. Flow cytometry was applied to measure and analyze the apoptosis rate of cells in each group. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) method was applied to detect the expression level of DUSP1 mRNA in cells. Methylation specific PCR (MSP) was applied to detect the methylation status of the DUSP1 promoter region in each group. Western blot was applied to analyze the expression levels of DUSP1 protein and key proteins in the DUSP1 downstream mitogen-activated protein kinase (MAPK) signaling pathway in each group. RESULTS: Compared with the Control group, the cell OD450 values (48 h, 72 h), scratch healing rate, number of cell invasions, and expression of DUSP1 in the OSM group were obviously decreased (P<0.05); the apoptosis rate, the methylation level of DUSP1, the expression of p38 MAPK protein, and the phosphorylation level of extracellular regulated protein kinases (ERK) were obviously increased (P<0.05); the cell OD450 values (48, 72 h), scratch healing rate, number of cell invasions, and expression of DUSP1 in the DAC group were obviously increased (P<0.05); the apoptosis rate, the expression of p38 MAPK protein, the phosphorylation level of ERK, and the methylation level of DUSP1 were obviously reduced (P<0.05). Compared with the OSM group, the cell OD450 values (48, 72 h), scratch healing rate, number of cell invasions, and expression of DUSP1 in the FA+OSM group were obviously decreased (P<0.05); the apoptosis rate, the methylation level of DUSP1, the expression of p38 MAPK protein, and the phosphorylation level of ERK were obviously increased (P<0.05). Compared with the FA+OSM group, the cell OD450 values (48, 72 h), scratch healing rate, number of cell invasions, and expression of DUSP1 in the FA+OSM+DAC group were obviously increased; the apoptosis rate, the methylation level of DUSP1, the expression of p38 MAPK protein, and the phosphorylation level of ERK were obviously reduced (P<0.05). CONCLUSIONS FA may inhibit DUSP1 expression by enhancing DUSP1 methylation, regulate downstream MAPK signal pathway, then promote apoptosis, inhibit cell invasion and metastasis, and ultimately reduce OSM resistance in NSCLC cells.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Dual Specificity Phosphatase 1/pharmacology* ; Cell Proliferation ; p38 Mitogen-Activated Protein Kinases/pharmacology* ; Methylation ; Apoptosis ; Cell Line, Tumor

Objective To understand the current situation of college students' knowledge, attitude and practice of new infectious diseases, to explore the influencing factors, and to provide references for the prevention and control of new infectious diseases among college students. Methods During the epidemic period, questionnaire on college students' knowledge, attitude and practice of new infectious diseases was used to conduct an online survey of college students nationwide. Results There were significant differences in the scores of knowledge among the students of different gender, major and province (F values were -8.26, 15.30, P < 0.01); The compliance rates of wearing masks and washing hands among the students of different gender were different (P < 0.01); The score of cognition level outside Hubei Province was higher than that in Hubei Province (P < 0.01); The compliance rates of wearing masks among the students with or without suspected cases were different (P < 0.05); There was a significant difference in the compliance rate of home quarantine between Hubei Province and other provinces (P < 0.01); There was a significant difference in the belief scores of college students of different majors (P < 0.01); The sources of Covid-19 knowledge included microblog, Wechat, QQ, and major forums, accounting for 95.1%, TV, radio, and newspaper, accounting for 89.5%, news website, accounting for 86.4%, family or friends, accounting for 64.50%, and community Village Committee propaganda, accounting for58.50%. College Students' knowledge and belief were positively correlated with behaviors (β = 0.079, 0.076, P < 0.05). Conclusion In view of the different characteristics of college students, different publicity plans should be formulated. The key to improve their prevention and control behavior is to strengthen the publicity of new infectious diseases knowledge and enhance their belief.

Objective To analyze the current status and trends of chronic kidney disease (CKD) burden in China from the global perspective, and to provide reference for the prevention and control of CKD in China. Methods The incidence, prevalence, mortality and disability adjusted life year (DALY) rate of China and other regions were retrieved from GBD 2019. Comparative analyses were carried out to reflect the burden of CKD. Results In 2019, the incidence and the prevalence in females were higher than those in males, while the mortality and the DALY rates in females were lower than those in males. There were obvious distinctions in different age groups and people over 65 years old had a higher CKD burden. The burden of CKD mainly came from hypertensive nephropathy and type 2 diabetic nephropathy. From 1990 to 2019, the DALY rate of CKD in China showed an overall upward trend. After eliminating the differences in age composition, the standardized DALY rate of CKD in China was higher than that in Japan, Korea and Ukraine although lower than the global. The main influencing factors of the burden of CKD were metabolic factors. Conclusion The burden of CKD in China is continuously rising. It is still necessary to pay attention to the three-level prevention and control, and focus on the key people groups and high-risk factors to further reduce the burden of CKD in China.

Objective To investigate the serum vitamin and trace element levels in children with short stature and their correlation with bone age. Methods Levels of serum VA and VD, and trace elements Ca, Fe, Zn, Mg, Cu, Pb and Cd were measured in 322 children who were referred for height consultation. Bone ages were evaluated and the correlation between bone age and serum vitamin and trace element levels was analyzed. Results The VA and VD deficiency rates of these 322 children were 22.05% and 34.16%, respectively. The deficiency rates of trace elements Ca, Fe and Zn were14.29%, 21.43% and 6.83%, respectively. The Pb excess rate was as high as 42.55%. The rates of bone age (BA) retardation in Group Ⅰ (short) and Group Ⅱ (slightly short) were 49.38% and 37.57%, respectively, which was significantly higher than that of Group Ⅲ (normal). The Ca level of BA retardation children was lower than that of the normal BA children in Group I. The VD level of BA retardation children was lower than that of the normal BA children in Group Ⅱ. BA was negatively correlated with VD, Ca, and Cu levels in children (r=-0.241; r=-0.136; r=-0.162), and positively correlated with Fe (r=0.286) . Conclusion There were significant abnormalities of vitamins and trace elements in short children. Children's bone age had a certain correlation with serum vitamin D, calcium, copper, and iron levels. Serum vitamin and trace element levels in children should be monitored to guide a reasonable diet to better promote child growth and development.

Objective To summary the hospitalization costs of lung cancer patients, and analyze the influence factors in these patients, and provide basis for controlling hospitalization costs of lung cancer patients. Methods The hospitalization costs data of hospitalized lung cancer cases in Wuhan from 2018 to 2020 were collected from medical records. Nonparametric test was used to analysis the data for single factor analysis. The patients were divided into two groups according on the upper quartile value of hospitalization cost, that is high-cost group (the cost ≥ the upper quartile value) and normal cost group (the cost “four major hospitals in Hubei^” respectively. The hospital type is an independent influencing factors, compared with specialized hospital, the OR is 4.726 for general hospital. The hospitalization days is the independent influencing factors, more hospitalization days has high cost. The treatment mode is the independent influencing factors, compared with non-operative treatment, the OR is 556.129, 18.156 and 5.212 for surgical model, radio therapy model and interventional model respectively. Conclusion The age, hospital level, hospital type, hospitalization days and treatment mode are the independent influencing factors of hospitalization costs. To reduce the hospitalization cost of lung cancer patients, we should standardize the diagnosis of lung cancer patients, and focus on standardizing the treatment mode, also considering other influencing factors, such as hospital level, hospital type.

Objective To investigate the effect of HQ on autophagy in human leukemia Jurkat T-cells and its mechanism. Methods (1) Exponentially growing Jurkat cells were cultured in vitro with HQ at the concentrations of 0 (control group), 12.5 and 25 mol·L^-1 for 24 hours. Western blotting was used to detect the protein levels of autophagy markers P62 and LC3II. Aggregations of LC3 fluorescent spots were determined by immunofluorescence technique. (2) Using the gene expression database, differentially expressed microRNAs (miRNAs) in CD34+ cells treated with HQ were selected to predict target genes through gene prediction tools such as TargetScan, miRBD and miRwalk. Enrichment analysis was conducted on these target genes to obtain relevant signal pathways. (3) Based on the results above, RT-PCR was used to examine the expression of Akt and Foxo3a genes in Akt/Foxo3a signal pathway, while western blotting was performed to determine protein expression levels of Akt, p-Akt, Foxo3a and p-Foxo3a in Jurkat cells exposed to HQ. Results (1) Compared with the control group, the expression of autophagy key protein P62 decreased in 25mol·L^-1 group, and the ratio of LC3II/LC3I increased in 12.5mol·L^-1 and 25mol·L^-1 groups (P<0.05). The results of immunofluorescence showed that the aggregation of LC3 fluorescent spots increased with the increase of HQ concentration. (2) Bioinformatic analysis revealed several autophagy-related signaling pathways including AMPK, MAPK, Hedgehog and PI3K/Akt signal pathways. (3) HQ treatment decreased gene expression levels of Akt and Foxo3a in Akt/Foxo3a signal pathway, and the protein levels of Akt, p-Akt, Foxo3a and p-Foxo3a were also significantly decreased in cells treated with HQ in a concentration dependent manner (P<0.05). Conclusion HQ may promote autophagy in Jurkat cells, which may be partly mediated through the inhibition of Akt/Foxo3a signal pathway..

OBJECTIVE To investigate the factors influencing the changes in purchasing quantity in the procurement varieties of the first batch of volume-based drug centralized procurement （hereinafter referred to as centralized procurement）. METHODS Using 25 procurement varieties of the “4+7” policy as research objects， the changes in purchasing quantity of procurement varieties were analyzed before and after the implementation of the “4+7” pilot， renewal and expansion policies. The influential factors were determined from the three levels of drugs， medical institutions and regions； and the multiple linear regression model was used to analyze the influential factors for the changes in the purchasing quantity of procurement varieties. RESULTS Before and after the implementation of the “4+7” pilot， renewal and expansion policies， the purchasing quantity increased by 52.1， －0.2， 85.8 ten thousand DDDs on average， compared with base period. During pilot， renewal and expansion period， DDDc decrease in procurement varieties was positively correlated with the increase in purchasing quantity （P＜0.01）. During the pilot and renewal period， the number of absolutely alternative varieties was positively correlated with the increase in purchasing quantity （P＜0.1）. During the pilot and expansion period， the number of alternative varieties to a certain extent was negatively correlated with the increase in purchasing quantity （P＜0.05）. During the renewal period， the increment of purchasing quantity in tertiary hospitals was smaller than that of primary medical institutions （P＜0.05）. CONCLUSIONS There is a relationship between the decline of DDDc and the changes in the purchasing quantity， that is， the more the drug price dropped， the more the purchasing quantity increased. The number of alternative varieties for centralized procurement will affect the changes in their purchasing quantity， but it is not always stable. With the implementation of the policy， the volume of primary medical institutions gradually exceeds that of tertiary institutions， indicating that the consumption of centralized purchased varieties is transferred to the primary medical institutions， and centralized procurement has promoted the implementation of the hierarchical diagnosis and treatment system.