Objective To prepare recombinant plasminogen activator(Pla) protein in E.coli BL21 cells that can be used in studying interactions between Yersinia pestis proteins and immunologic diagnosis of plague.Methods The pla gene was amplified by PCR and cloned into the pET28a expression vector.E.coli BL21 competent cells were transformed with the recombinant vectors, and isopropyl-β-D-thiogalactopyranoside ( IPTG) was added to induce expression of Pla protein. The expressed protein was detected by SDS-PAGE electrophoresis.The inclusion bodies of Pla protein were denatured in 8 mol/L urea, and then refolded using gradient urea solutions.The purified protein was identified by SDS-PAGE electrophoresis and Western blot.Results and Conclusion The constructed expression vector was demonstrated to be correct through agarose gel electrophoresis and sequencing.The recombinant Pla protein was accumulated as an inclusion body in E.coli, and the overexpression product was mainly a target protein, the yield of which was very high.SDS-PAGE purity of the bioactive Pla protein was obtained by denaturing and refolding the inclusion bodies.This study provides a simple and quick method for highly efficient preparation of biologically active Pla protein.

Objective To establish a simple and quick loop-mediated isothermal amplification (LAMP)method for detection of Yersinia pestis.Methods LAMP Primers were designed based on the specific sequence 3a in Y.pestis chromosome.LAMP reaction results were detected using turbidity meter or visual method.The specificity of the constructed method was evaluated by detecting Y.pestis and its closely-related bacteria.The different dilution DNA template was detected with LAMP and PCR to evaluate the sensitivity of the method.Results Thirty strains of bacteria closely related to Y.pestis were detected by the constructed LAMP,and all the results were negative,indicating that the method had a very high specificity.The detection sensitivity of this LAMP assay was 20 pg of DNA per reaction,which was ten-fold that of the regular PCR.The detection reaction was completed in 25 min.Conclusion This LAMP method is quick,sensitive, specific and simple,which is expecked to become an effective method for rapid detection of Y.pestis on the scene.

In response to the major national strategic needs of "Healthy China" and "Chengdu-Chongqing Economic Circle", and in strict accordance with the requirements of "New Medicine" construction, Chongqing University builds the medical discipline with a high starting point, develops high-quality medical education, and promotes the exploration and practice of elite education of clinical medical talents in the cross-disciplinary context of medicine and engineering. From the perspective of SWOT, this study analyzed the situations and trends of clinical medical education in "Double First Class" universities which newly established the medical discipline. Combined with the training model of clinical talent class in Chongqing University and the practices of "Four Capacity-Buildings", "Four Integration-Measures", and "Three Optimization-Mechanisms", we discuss the thoughts and strategies of cultivating clinical medical talents in the background of "New Medicine" construction and in the cross-disciplinary context of medicine and engineering.

Objective To explore the effect of quality control circles (QCCs) in improving the delivery health checkup report.Methods QCC was founded with the theme of"improving the quality of health checkup report delivery."First,we planned an activity schedule and identified topics.We then set target focuses for service personnel,distribution modes,and operating environments;planned countermeasures;and selected optimal policies.Circle members implemented the optimal policies jointly.Reports of physical examinations by the Guoyu health management center were selected and analyzed.The total number of reports before improvement (January to December 2015) was 59 189 of which 34 549 (58.4％) were male patients and 24 640 (41.6％) were female patients;their average age was (37.7± 11.4) years.The total number of reports after improvement (December 2016 to January 2017) was 6 568,of which 3 881 (59.1％) were male patients and 2 687(40.9％) were female patients;their average age was (39.9± 11.7) years.We compared the quality indicators and evaluated the comprehensive quality of the patients before and after improvement.A total of 65 531 physical examination reports of subjects examined at the center between February and December 2017 were selected for effect tracking,including 39 230 (59.9％) men and 26 301(40.1％) women,aged (38.1±11.5).Results The on-time delivery rate of the health examination reports from rose from 51.4％ to 94.0％.The ratio of system leakage to sign for reports decreased from 14.5％ to 0.8％.The average time between the examination and when each report was handed over to for distribution decreased from 29.8 hours to 4.2 hours,and the average time between each report being distributed to the providers checking in dropped from 509.8 hours to 72.8 hours,while the average time for the preparation of each report for delivery decreased from 13.5 seconds to 3.1 seconds.The average time between delivery of a report and its being signed decreased from 4.3 seconds to 0.1 seconds.Before the improvement,the expected goals were not met.After improvement,the delivery rate of the health examination reports was 100.0％,the delivery intact rate of the group reports was 100.0％,and the satisfaction rate of the group reports was 99.4％.The comprehensive quality for the members was obviously higher after the improvement than before.After 11 months of tracking,the delivery accuracy rate of health examination report still failed to reach the target value of 100.0％,but all other indicators reached the target value,with good results.Conclusions Application of QCC not only improved the delivery the health checkup reports,but also promoted service quality after medical examinations and ended medical dispute caused by the loss of physical examination reports.

Objective:To observe the effect of Yi Jin Jing(Sinew-transforming Qigong Exercises)intervention on anxiety in asymptomatic patients with Corona Virus Disease 2019(COVID-19)infection during quarantine.Methods:A total of 160 asymptomatic patients with COVID-19 infection were stratified by gender and divided into an observation group and a control group by the stratified randomization method,with 80 cases in each group.The control group was given basic nursing in the cabin,and the observation group was given additional Yi Jin Jing exercises once a day,20 min each time,and trained continuously until 1 d before leaving the cabin.The Hamilton anxiety scale(HAMA)score was compared between the two groups.Results:A total of 154 cases were included for data analysis in this study,including 74 cases in the observation group and 80 cases in the control group.After intervention,the HAMA scores in both groups increased(P<0.05),while the score in the observation group was lower than that in the control group(P<0.05).In the control group,the HAMA score of females after intervention was higher than that of males.The scores of mental anxiety and somatic anxiety in both groups were higher than those before intervention,while the score of mental anxiety in the observation group was significantly lower than that in the control(P<0.05).However,there was no statistical difference in the somatic score between the two groups(P>0.05).Conclusion:Patients with COVID-19 infection will get anxious during quarantine.Yi Jin Jing exercise can effectively reduce the aggravation of anxiety in asymptomatic patients with COVID-19 infection during concentrated quarantine.

Objective: To investigate the characteristics, temporal trend of silicosis, and provide basis for risk assessment and precise prevention and control of occupational diseases. Methods: Using descriptive statistics to analyze the reported cases of silicosis by SPSS 20.0 software. Reported silicosis cases, the constituent ratio, the incidence age and the working age at onset were analyzed by a linear trend test. Analyzing the variation trends of regional, industry, economic type and enterprise scale distributions by the chi-square trend test. Moreover, using Moran's I method for spatial autocorrelation analysis and trend-surface analysis. Results: (1) During 2006 to 2015, Guangdong province had reported 1, 428 cases of silicosis, mainly gathered in Foshan, Zhongshan, Guangzhou, Shenzhen, which included 1391 male cases accounting for 97.41%. And the average incidence age was 45 (39, 51) . The average working age of onset was 9 (5.5, 15) . In economic type distribution, the private economy took the main part, accounting for 59.1%. In enterprise scale distribution, it was dominated by small and medium enterprises (SMEs) , accounting for 32.4% and 37.3% respectively. In industry distribution, most cases were gathered in materials and mining industry, accounting for 32.1% and 22.9% respectively. (2) The number of silicosis cases, the incidence age and the working age of onset showed a rising trend (P<0.01) . Meanwhile, the constituent ratios of medium-sized enterprises and building materials industry were increasing (P<0.05) . The annual variation trends of regional, economic type and age distributions were not statistically significant (P> 0.05) . (3) The spatial distribution trend showed an inverted U type, which was firstly raised and then declined from south to north and from east to west. The distribution characteristic demonstrated some high-high cluster areas, including Chancheng, Nanhai, Shunde, Panyu, Dongguan, Pengjiang, and Zhongshan. While Wuhua showed a high-low outlier form (P<0.01) . Conclusion Silicosis cases, age and working age of onset were on the rise, as well as the industry and enterprise scale distributions of occupational diseases presented a certain trend in Guangdong province from 2006 to 2015. There were high-high cluster and high-low outlier phenomena in spatial distribution with spatial correlation. Therefore, our work of silicosis epidemic trend and distribution may provide some bases for the occupational disease risk assessment and control.

Objective:To diagnose and explore the genetic etiology of a neonate with Hereditary epidermolysis bullosa.Methods:A neonate who was admitted to Suqian Hospital Affiliated to Xuzhou Medical University on July 10, 2021 was selected as the study subject. Peripheral blood samples were collected from the child and his parents for the extraction of genomic DNA. And target gene capture and next-generation sequencing were carried out. Candidate variants were verified by Sanger sequencing and pathogenicity analysis.Results:The child was found to harbor compound heterozygous variants of the COL17A1 gene, namely c. 997C>T (p.Q333X) and c. 3481dupT (p.Y1161fs*2), which were respectively inherited from his father and mother. Both variants were predicted to be pathogenic. Conclusion:The child was diagnosed with Generalized atrophic benign epidermolysis bullosa due to the compound heterozygous variants of the COL17A1 gene.

OBJECTIVE: To analyze the epidemiological characteristics and predict epidemiological trends of occupational chemical poisoning,based on directly reported data during 2006-2015 in Guangdong Province. METHODS: The data of patients with occupational chemical poisoning reported from National Information Surveillance System for Occupational Disease and Occupational Health from 2006 to 2015 in Guangdong Province were collected. The epidemiological characteristics were retrospectively analyzed. The autoregressive integral moving average model( ARIMA model) was established and validated based on the number of the new onset cases and was used to predict the trends of occupational chemical poisoning from 2017 to 2020 in Guangdong Province. RESULTS: From 2006 to 2015,1 288 new cases of occupational chemical poisoning were reported in Guangdong Province,which accounted for 24. 4% of the total number of new cases of occupational diseases in the province( 5 283 cases). Among the new cases,the percentage of acute and chronic poisoning was 21. 7%( 279/1 288) and 78. 3%( 1 009/1 288). There was 74. 7%( 962/1 288) of organic solvent poisoning. Five kinds of new occupational chemical poisoning were found. Most of the new cases were male,accounting for 56. 7%( 729/1 288). They were mainly distributed and concentrated in Pearl River Delta Region,accounting for 95. 9%(1 235/1 288). Shenzhen,Dongguan and Guangzhou were the most three cities which had 425,325 and 209 cases respectively,all of them accounted for 74. 4%( 959/1 288). The new cases of poisoning mainly distributed in medium and small enterprises( 72. 0%),private economic enterprises( 50. 9%) and manufacturing industries(70. 5%). The number of occupational chemical poisoning diseases decreased first,and increased,and the proportion to the total number of occupational diseases in Guangdong Province showed a straight downward trend(P < 0. 01). The median age at diagnosis was 35 years old and the median work year at diagnosis was 2. 0 years,and both of them showed an increasing trend( P < 0. 01). CONCLUSION: Occupational chemical poisoning in Guangdong Province has certain characteristic of crowd aggregation and epidemic trends.

Objective: To investigate the feasibility of long noncoding RNA (lncRNA)_AK096792 as a clinical predictor of bronchopulmonary dysplasia (BPD) in preterm infants. Methods: All the cord blood(2-5 mL) of very low birth weight (VLBW) preterm infants born in Huai′an First Hospital Affiliated to Nanjing Medical University were collected from December 1, 2015 to December 1, 2017.Moreover, the peripheral blood(2 mL) of those VLBW infants diagnosed with BPD was also collected.A total of 36 infants with BPD were collected.Another 36 cases of premature children with VLBW were chosen as control group according to random number table.The relative content of lncRNA_AK096792 in cord blood and peripheral blood was detected by using real-time quantitative PCR (qPCR). Additionally, the correlation of lncRNA_AK096792 levels between cord and peripheral blood of BPD infants was analyzed.The sensitivity and specificity of lncRNA_AK096792 for BPD were analyzed by using receiver operating curve test. Results: (1)LncRNA_AK096792 was a common, evolutionarily conserved, non-coding RNA present in both mouse and human.(2) The expression level of lncRNA_AK096792 in peripheral blood was significantly higher than that in cord blood in BPD group[(463.3±352.0)% vs.(50.0±37.5)%], and the difference was significant(P<0.001), and they were highly correlated (r=0.825, P<0.001). (3) The level of lncRNA_AK096792 in cord blood in BPD group was signi-ficantly higher than that in non-BPD group [(484.3±280.5)% vs.(101.2±28.6)%], and the difference was significant(P<0.001). (4)When lncRNA_AK096792 served as a clinical predictor for BPD, the specificity was 83.3%, the sensitivity was 75.6%, and an area under the receiver operating characteristic curve was 0.88(P<0.001). Conclusions LncRNA_AK096792 is highly correlated with the development of BPD.The level of lncRNA_AK096792 in umbilical cord blood of premature infants can be used as an early predictive marker for BPD, it calls for further study.

As a novel and promising antitumor target, AXL plays an important role in tumor growth, metastasis, immunosuppression and drug resistance of various malignancies, which has attracted extensive research interest in recent years. In this study, by employing the structure-based drug design and bioisosterism strategies, we designed and synthesized in total 54 novel AXL inhibitors featuring a fused-pyrazolone carboxamide scaffold, of which up to 20 compounds exhibited excellent AXL kinase and BaF3/TEL-AXL cell viability inhibitions. Notably, compound 59 showed a desirable AXL kinase inhibitory activity (IC₅₀: 3.5 nmol/L) as well as good kinase selectivity, and it effectively blocked the cellular AXL signaling. In turn, compound 59 could potently inhibit BaF3/TEL-AXL cell viability (IC₅₀: 1.5 nmol/L) and significantly suppress GAS6/AXL-mediated cancer cell invasion, migration and wound healing at the nanomolar level. More importantly, compound 59 oral administration showed good pharmacokinetic profile and in vivo antitumor efficiency, in which we observed significant AXL phosphorylation suppression, and its antitumor efficacy at 20 mg/kg (qd) was comparable to that of BGB324 at 50 mg/kg (bid), the most advanced AXL inhibitor. Taken together, this work provided a valuable lead compound as a potential AXL inhibitor for the further antitumor drug development.