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Objective:To observe the inhibition of SARS-CoV-2 spike protein (S-protein) on the proliferation of human retinal pigment epithelium (RPE) cells.Methods:SARS-CoV-2 S-protein gene fragment expression plasmid (p3xflag-S) was constructed and transfected into human RPE, HEK293 cells. DNA sequencing was used for identification, and the expression of Flag-S was detected by Western blot. HEK293 cells were divided into the cells 1, 2, 3 and 4 and transfected with GFP11 plasmid and vector, GFP1-10 plasmid and vector, transfected with GFP11 and pCMV-HA-ACE2 plasmid, GFP1-10 and p3xflag-S plasmid. Cell 1 was co-cultured with cell 2 (control group 1), cell 2 with cell 3 (control group 2), cell 3 with cell 4 (observation group), and cell 1 mixed with cells 2, 3 and 4 (control group 3). Bright-field microscopy and fluorescence microscopy were used to observe cell fusion. RPE cells were divided into control group and overexpression S-protein group. The cell cycle was detected by flow cytometry; the cell proliferation level was detected by Counting Kit 8 (CCK-8); and the S-protein expression level in RPE cells was detected by Western blot. The Student’s t-test was performed for comparison between groups. Results:DNA sequence assay showed that S-protein cDNA was fused with flag-tagged protein. Western blot assay showed that S-protein-related expression was elevated in transfected HEK293 cells compared with untransfected p3xflag-S cells. Large, multinucleated fused cell clusters were visible under bright-field microscopy; multiple nuclear with distinct green fluorescence were visible in the fused cells under fluorescence microscopy. Western blot assay showed elevated S-protein-related expression in transfected p3xflag-S plasmid RPE cells compared to untransfected p3xflag-S plasmid RPE cells. CCK-8 results showed that the proliferative capacity of RPE cells in the S-protein overexpression group was significantly reduced compared with the control group, with statistically significant differences ( t=22.70, 16.75, 23.38; P<0.000 1). The results of flow cytometry showed that the G1 phase cells in the control and overexpression S-protein groups were 41.1 % and 67.0%, respectively; compared with the control group, the G1 phase cells in the overexpression S-protein group were significantly higher, and the difference was statistically significant ( t=4.76, P=0.018). The apoptosis rate was significantly increased in the S-protein overexpression group compared with the control group, and the difference was statistically significant ( t=4.91, P=0.008). Conclusion:Overexpression of the SARS-CoV-2 spike protein reduced the proliferation of human RPE cells.
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Objective To sort out the scientific research achievements in the direction of early warning of infectious diseases in China from 2001 to 2022, and analyze the research hotspots and trends in the direction of early warning of infectious diseases in China in recent 20 years, so as to provide reference for relevant policies and exploration directions. Methods Relevant literature retrieved from CNKI Chinese database was used as the data source, and Excel 2019 and Citespace 6.1.R2 software were used for visual analysis of research hotspots and frontier literature. Results A total of 1276 papers meeting requirements were obtained, and most of the research groups were relatively small and had little cooperation with others. The types of research institutions were relatively single, and most of them were domestic universities. “Infectious diseases”, “early warning” and “prediction” were the most frequently used keywords. Research on big data and COVID-19 epidemic prevention and control is the current research frontier. Conclusion There is little cooperation among authors and between institutions in the field of early warning of infectious diseases in China. Using big data to early warning of infectious diseases and improving the ability of early warning of COVID-19 are the main research directions and trends at present.
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Objective:To explore the inhibitory effect of long non-coding RNA (lncRNA) KCNQ1 overlapping transcript 1 (KCNQ1OT1) by targeting microRNA-199a-5p (miR-199a-5p) on the apoptosis of human lens epithelial cells (LECs).Methods:The anterior lens capsule tissue of 23 age-related cataract patients who underwent cataract surgery in Xinxiang First People's Hospital from December 2018 to August 2019 was collected.At the same time, anterior lens capsules from 20 healthy donor were collected.The expressions of KCNQ1OT1 and miR-199a-5p in the tissues were detected by real-time fluorescence PCR.Human LECs SRA01/04 cultured in vitro were divided into blank control group, model control group, small interfering RNA-negative control (siR-NC) group, siR-KCNQ1OT1 group, miR-NC group, miR-199a-5p group, siR-KCNQ1OT1+ anti-miR-NC group and siR-KCNQ1OT1+ anti-miR-199a-5p group.No intervention was administered to blank control group.Cells in model control group were cultured with 100 μmol/L H 2O 2 for 24 hours to establish oxidative stress injured model, and cells in the other six groups were transfected with corresponding transfection reagents for 6 hours by liposome method according to grouping, and then treated with 100 μmol/L H 2O 2 for 24 hours.The expressions of KCNQ1OT1 and miR-199a-5p in lens anterior capsule tissue and LECs cells were determined by real-time fluorescent quantitative PCR.Cell viability was detected with thiazolyl blue (MTT). Cell apoptosis was analyzed by flow cytometry.The expressions of B-cell lymphoma/leukemia-2 (bcl-2) and bcl-2 related X protein (Bax) proteins were assayed by Western blot.The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by enzyme-linked immunosorbent assay (ELISA). The targeting relationship between KCNQ1OT1 and miR-199a-5p was verified by dual luciferase reporter experiment.The study protocol was approved by an Ethics Committee of Xinxiang First People's Hospital (No.2019-001). Written informed consent was obtained from relatives of patient. Results:The relative expression of KCNQ1OT1 in the anterior capsule of patients with age-related cataract was 2.41±0.42, which was significantly higher than 0.97±0.19 of normal people, and the relative expression of miR-199a-5p in the capsule of patients with age-related cataract was 0.36±0.12, which was lower than 1.04±0.15 of normal people, and the differences were statistically significant ( t=14.112, 16.507; both at P<0.001). Compared with blank control group, the relative expressions of KCNQ1OT1 and bax protein, cell apoptosis rate and MDA content were significantly increased, and the relative expressions of miR-199a-5p and bcl-2 protein, cell viability and SOD activity were significantly reduced in model control group, showing statistically significant differences (all at P<0.001). Compared with siR-NC group, the relative expressions of KCNQ1OT1 and bax protein, cell apoptosis rate and MDA content in cells of siR-KCNQ1OT1 group were decreased, while the relative expression of bcl-2 protein, cell survival rate and SOD activity were increased, and the differences were statistically significant (all at P<0.05). Compared with miR-NC group, the KCNQ1OT1-wild type (WT) luciferase activity in miR-199a-5p group was significantly decreased, with a statistically significant difference ( t=21.131, P<0.001). The relative expression levels of miR-199a-5p and bcl-2 proteins, cell survival rate and SOD activity were significantly increased, and the relative expression of bax protein, cell apoptosis rate and MDA content were significantly decreased in miR-199a-5p group than those in miR-NC group, and the differences were statistically significant (all at P<0.05). The relative expression levels of miR-199a-5p and bcl-2 proteins, cell survival rate and SOD activity were significantly lower, and the cell apoptosis rate, relative expression of bax protein and MDA content were significantly higher in siR-KCNQ1OT1+ anti-miR-199a-5p group than those in siR-KCNQ1OT1+ anti-miR-NC group, and the differences were statistically significant (all at P<0.05). Conclusions:The inhibition of KCNQ1OT1 can promote the cell viability of human LECs, inhibit H 2O 2-induced cell apoptosis and oxidative stress, and the mechanism may be related to the up-regulation of miR-199a-5p.
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Objective@#To explore the prevalence of sexting among secondary vocational school students and to examine the correlation between sexting and mental health, so as to provide evidence for prevention and intervention on adolescents mental health.@*Methods@#In April 2021, three secondary vocational schools from central urban area, outskirts and outer suburbs in Shanghai, among 1 580 students were selected by cluster sampling.@*Results@#The proportions of students who received and who forwarded others private photos or videos were 11.96% and 2.78%, respectively. The proportion of students who sent others personal photos or videos was 2.54%. And the proportions of boys were higher than those of girls ( χ 2=45.89, 24.09, 6.66, P <0.01). The results of multivariable Logistic analysis showed that compared with those who had never sexted, students who had ever had any sexting were at higher risk of anxiety ( OR=2.40, 95%CI =1.59-3.61), depression( OR=1.86, 95%CI =1.25-2.75), non suicidal self injury intention ( OR=2.34, 95%CI =1.48-3.69) and suicidal intention ( OR=2.96, 95%CI =1.83-4.79)( P < 0.05 ).@*Conclusion@#There is a significant correlation between sexting and mental health among secondary vocational school students. The latent risks of sexting should be included in the curriculum of comprehensive sex education for teenagers to enhance their media literacy and decision making capability, so as to prevent sexual assault or abuse caused by sexting and promote sexual and reproductive health and as well as mental health of adolescents.
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To investigate the effectiveness and safety of diverticular peroral endoscopic myotomy (D-POEM) for mid-esophageal diverticulum. Data of consecutive patients in the prospective database with mid-esophageal diverticulum who received D-POEM in West China Hospital, Sichuan University between April 2014 to September 2019 were collected. The modified Eckardt scoring system for mid-esophageal diverticulum was used to evaluate the severity of diverticular symptoms. The effectiveness and safety of D-POEM were evaluated in terms of clinical success, technical success, complications and recurrence. A total of 7 patients with mid-esophageal diverticulum were included. Clinical and technical success was achieved in all patients with operation time of 16-70 minutes. No serious complications (2 cases with minor complications) or recurrence occurred. The follow-up time was 2-16 months. The median modified Eckardt score decreased from 3 points before operation to 0 points after operation. It is preliminarily believed that D-POEM ensures a complete septotomy between normal esophageal lumen and esophageal diverticulum. It is a safe and effective technique for mid-esophageal diverticulum. The modified Eckardt scoring system is suitable for symptom evaluation before and after treatment of mid-esophageal diverticulum.
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Flavonoid glycosides are the main active constituents of Epimedii Folium and its related plants. They can be divided into polyglycosides and low glycosides according to the number of glycosyl group. The polyglycosides of Epimedii Folium can be transformed into low glycosides after biotransformation ；pharmacological activities of low glycosides in anti-tumor ，tonifying kidney yang and anti-osteoporosis are stronger than those of polyglycosides. In this paper ， the research progress about biotransformation technology of flavonoid glycosides of Epimedii Folium was reviewed. It was found that the main biotransformation pathway of flavonoid glycosides of Epimedii Folium was to obtain low glycosides by removing glycosyl group ； related methods were mainly enzymatic hydrolysis and microbial transformation ，and also included plant cell transformation ，acid hydrolysis method and synthesis method.
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OBJECTIVE To prepare hyperoside mixed nanomicelles （Hyp-F127/TPGS） and optimize its preparation technology，and to investigate its intestinal absorption characteristics. METHODS Hyp-F127/TPGS was prepared by thin film dispersion method. Based on single factor test and Plackett-Burman design ，combined with Box-Behnken response surface method ， the preparation process was optimized and validated using entrapped efficiency （EE）and drug loading （DL）as evaluation indexes ， F127-TPGS mass ratio ，hydration time and the amount of Hyp as factors. The appearance and microscopic morphology of Hyp-F127/TPGS obtained by the optimal technology were observed ，and the particle size ，polydispersity index （PDI）and Zeta potential were also determined. The critical micelle concentration （CMC）of blank micelle （F127/TPGS），in vitro release behavior and preliminary stability of Hyp-F 127/TPGS were investigated ，and absorption characteristics of Hyp-F 127/TPGS were investigated by in situ unidirectional intestinal perfusion model. RESULTS The optimal preparation technology of Hyp-F 127/TPGS included F127-TPGS mass ratio of 2∶1，hydration time of 2 h，and Hyp amount of 9 mg. Results of three validation tests showed that the EE of Hyp-F 127/TPGS was （87.20±0.99）％，and the DL was （5.02±1.20）％，deviations from predicted values were 0.92％ and 2.39％. The micelles prepared by optimal technology were yellow ，clear and transparent solution ，with good Tyndall effect ；under transmission electron microscope ，they were spherical ，complete and evenly distributed ；the particle size was （15.02±0.16）nm， the PDI was 0.092±0.031，and the Zeta potential was （－6.67±1.47）mV. The CMC of F 127/TPGS was 21 μg/mL，Hyp-F127/ TPGS was stable after 4 weeks of storage at 4 ℃，and the cumulative release rates of Hyp-F 127/TPGS and Hyp control were （66.30±2.93）％（96 h）and（99.24±0.27）％（60 h），respectively. Hyp-F 127/TPGS and Hyp reference were absorbed in each intestinal segment ，and the main absorption sites were jejunum and duodenum respectively ；drug absorption rate constant andapparent absorption coefficient of the former were significantly higher than those of the latter （P＜0.05 or P＜0.01）. E-mail：zhangyuhangxz@163.com CONCLUSIONS The optimized preparation technology of Hyp-F127/TPGS is stable and feasible ；prepared Hyp-F 127/ TPGS shows a sustained -release effect ，which promotes the intestinal absorption of H yp to a certain extent.
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We report a fetus with ectopic connection of venous catheter into the dilated coronary sinus, right-sided aortic arch, left-sided ductal arch, and aberrant left subclavian artery detected by routine prenatal ultrasound screening at 23 +2 gestational weeks. The baby was born vaginally at 38 +2 gestational weeks with an Apgar score of 10 at both 1 and 5 min. The reexamination of neonatal echocardiography on the second day after birth showed dilation of the internal diameter of the coronary sinus, right-sided aortic arch, and aberrant left subclavian artery. Follow-up at 90 days after birth found no abnormal growth and development.
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@#Esophageal diseases include esophageal malignant diseases and benign diseases, with a high incidence in our country. Along with the development of the endoscopic technique, many of them which required medical treatment or surgery in the past can now be cured by endoscopic surgery. This article is an overview of long-term follow-up of endoscopic surgery for the common esophageal disease, such as early squamous cell carcinoma, esophageal stricture, achalasia and submucosal tumor of the esophagus.
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@#Endoscopic resection and surgical resection are the two major therapeutic methods for early esophageal cancer. Endoscopic resection is safe and minimally invasive, but lymph node dissection can not be performed. Although surgery provides a rather thorough resection of the lesions and affected lymph nodes, surgical trauma brings certain negative impact on patients' long-term life quality. A comprehensive assessment of the patient's general condition, the risk of diseased lymph node metastasis, and the risk of the treatment itself is an important measure to optimize treatment decisions and formulate personalized treatment plans.
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Stem cells are of great interest to the scientific community due to their potential role in regenerative and rejuvenative medicine. However, their role in the aging process and carcinogenesis remains unclear. Because DNA replication in stem cells may contribute to the background mutation rate and thereby to cancer, reducing proliferation and establishing a relatively quiescent stem cell compartment has been hypothesized to limit DNA replication-associated mutagenesis. On the other hand, as the main function of stem cells is to provide daughter cells to build and maintain tissues, the idea of a quiescent stem cell compartment appears counterintuitive. Intriguing observations in mice have led to the idea of separated stem cell compartments that consist of cells with different proliferative activity. Some epithelia of short-lived rodents appear to lack quiescent stem cells. Comparing stem cells of different species and different organs (comparative stem cell biology) may allow us to elucidate the evolutionary pressures such as the balance between cancer and longevity that govern stem cell biology (evolutionary stem cell biology). The oral mucosa and its stem cells are an exciting model system to explore the characteristics of quiescent stem cells that have eluded biologists for decades.
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Objective To evaluate appropriate timing of laparoscopic cholecystectomy (LC) after percutaneous transhepatie gallbladder drainage (PTGD) for the treatment of acute cholecystitis (AC).Methods To compare the clinical data of 74 patients with acute cholecystitis who underwent laparoscopic cholecystectomy at different time after PTGD,Including the degree of gallbladder inflammation,operation time,intraoperative blood loss,the rate of conversion to laparotomy,length of stay and total cost.74 cases were divided into 3 groups,group 1 (n =31),operated within 4 w,group 2 (n =22),between 4 and 8 w,group 3 (n =21),after 8 w.Results Group 2 was better than group 1 and group 3 in operation time,blood loss,length of stay and total cost,the difference was statistically significant (P ＜ 0.05).Conclusion Laparoscopic cholecystectomy performed after 4-8 weeks PTGD is the best time in terms of less risk of operation,shorter hospital stay,lower cost,and better quality of life.
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Objective To study the protective effect of lycopene on vascular endothelial cell injury by cigarette smoke extract ( CSE) . Methods CSE was prepared and the human umbilical vein endothelial cells ( HUVECs) were assigned into four groups, cells in control were untreated, and cells in other three groups were treated by 10%CSE, 10%CSE+1. 0 μmol·L-1 lycopene and 1. 0 μmol·L-1 lycopene, respectively. Cell viability was evaluated using MTT assay. The intracelluar reactive oxygen species ( ROS) level was detected by ROS assay kits. Cell cycle and apoptosis were analyzed by flow cytometry. SIRT1 expression was detected by real-time fluorescence quantification PCR ( qRT-PCR) and Western blot. Results Cell viability in 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene or 1. 0 μmol·L-1 lycopene group was (56. 7±5. 1)%,(75. 6±7. 1)% and (95. 5± 9. 7)%, respectively. ROS assay showed that the relative fluorescence intensity in the control was 25. 3±3. 9, however, in 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene group or 1. 0 μmol·L-1 lycopene group were 67. 3±4. 6, 45. 3±3. 9 and 20. 8±2. 9, respectively. 10%CSE could induce G2 arrested and which could be antagonized by 1. 0 μmol·L-1 lycopene. The apoptosis rate in the control, 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene group or 1. 0 μmol·L-1 lycopene group was (6. 2±0. 5)%, (30.8±4.3)%, (18. 3±1. 9)% and (7. 6±0. 4)%, respectively. As shown in qRT-PCT testing, compared with the control, mRNA of SIRT1 in 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene group and 1. 0 μmol·L-1 lycopene group was (0. 51± 0. 03) fold, (0. 84±0. 05) fold, and (1. 31±0. 08) fold compared to the control, the data from western blot were consistent with qRT-PCR results. Conclusion Lycopene can prevent HUVECs from injury induced by CSE by upregulation of SIRT1.