Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Objective To investigate the pathogenic distribution characteristics and drug resistance of opportunistic Candida in patients with pulmonary tuberculosis on the Qinghai-Tibet Plateau.Methods 3 012 hospitalized cases of pulmonary tuberculosis at Qinghai Province Fourth People's Hospital from March 1,2020 to December 31,2020 were analyzed,sputum samples were collected,Candida identification was carried by VITEK-32-YBC automatic bacterial analysis system,and the detected Candida was tested for drug sensitivity.Results Among the 3 012 cases of pulmonary tuberculosis in this investigation,283 cases of pulmonary tuberculosis patients with Candida infection,accounting for 9.40%.Among them,Candida albicans was the main type of Candida,accounting for 79.86%of the total.Conclusion The prevalence rate of pulmonary tuberculosis complicated with Candida infection was high in Qinghai-Tibet Plateau.Therefore,the selection of antimicrobial drugs should be based on a comprehensive analysis of the patient's condition,in order to select the best and most effective drugs for treatment.

[摘 要] 目的：探究SOX9通过上调长链非编码RNA LINC01503的表达对喉鳞状细胞癌（LSCC）细胞的增殖、迁移、侵袭及肿瘤干细胞干性的影响。方法： 常规培养人LSCC细胞AMC-HN-8、TU177、TU212和TU686，用转染试剂将敲减序列及其对照核酸（si-SOX9-NC、si-SOX9#1、 si-SOX9#2、si-LINC01503-NC、si-LINC01503#1、si-LINC01503#2）或过表达质粒及其对照核酸（pcDNA3.1-SOX-NC、pcDNA3.1-SOX-oe、pcDNA3.1-LIN01503-NC和pcDNA3.1-LIN01503-oe）分别转染至TU177细胞或TU686细胞，记为si-SOX9-NC组、si-SOX9#1组、si-SOX9#2组、si-LINC01503-NC组、si-LINC01503#1组、si-LINC01503#2组；pcDNA3.1-SOX9-NC组、pcDNA3.1-SOX9-oe组、pcDNA3.1-LINC01503-NC组、pcDNA3.1-LINC01503-oe组、si-SOX9-NC + pcDNA3.1-LINC01503-NC组和si-SOX9 + pcDNA3.1-LINC01503-oe组。qPCR法检测SOX9 mRNA和LINC01503 在各组细胞中的表达，生物信息学分析SOX9与LINC0503启动子区的结合位点，双萤光素酶报告基因实验和染色质免疫共沉淀实验验证SOX9与LINC01503启动子区是否直接结合，WB法检测SOX9的敲减效率及LINC01503对TU177和TU686细胞干性标志物表达的影响，MTS法检测各组细胞的增殖活力，划痕愈合和Transwell小室实验检测各组细胞的迁移能力，克隆形成实验检测各组细胞的克隆形成能力。结果：SOX9在各种LSCC细胞中呈高表达（均P < 0.05），数据库数据分析显示，在头颈部鳞状细胞癌中，SOX9与LINC01503表达呈正相关（R = 0.12，P = 0.005 9）；SOX9可与LINC01503启动子区直接结合并促进其转录表达（均P < 0.05）；敲减LINC01503可明显抑制TU177细胞的增殖、迁移、侵袭（均P < 0.05），过表达LINC01503明显促进TU686细胞增殖、迁移、侵袭的能力（均P < 0.05），提高TU686细胞克隆形成能力和细胞干性标志物分子CD133、OCT4、SOX2的mRNA和蛋白水平表达（均P < 0.05），敲减LINC01503则均可抑制TU686细胞的克隆形成和细胞干性标志物的表达（均P < 0.05）；敲减SOX9均可明显抑制TU177细胞的增殖、迁移和侵袭能力，降低其干性细胞标志物的表达（均P < 0.05），同时过表达LINC01503则可部分逆转敲减SOX9对TU177细胞恶性生物学行为和干性标志物表达的抑制作用（均P < 0.05）。结论：SOX9和LINC01503在LSCC细胞中呈高表达，SOX9可能通过上调LINC01503表达提高LSCC细胞增殖、转移和侵袭能力和肿瘤干细胞干性。

Objective To analyze the effect of microRNA-22-3p (miR-22-3p) of extracellular vesicles derived from bone marrow mesenchymal stem cells on premature ovarian failure. Methods Follicular fluids were provided by premature ovarian failure patients with in vitro fertilization or the second-generation in vitro fertilization treatment and normal volunteers with the same treatment for infertility due to male factors; the exosomes derived from bone marrow mesenchymal stem cells were isolated by differential centrifugation; the morphology, particle size and marker proteins of isolated exosomes were analyzed by transmission electron microscopy, NanoSight LM10 analyzer and Western blotting. Granulosa cells were treated with cyclophosphamide (CTX), exosomes, miR-22-3 mimics and corresponding controls, and were divided into CTX group, control group, exosome incubation group, PBS treatment group, CTX+miR-22-3p group, CTX+miR-NC group, CTX+Exosomes group, and CTX+PBS group; gene expression was detected by Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR). The 3-(4, 5-Dimethylthazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reagent and flow cytometry were used to detect cell viability and apoptosis. Results The extracted exosomes had typical goblet vesicles, the exosome marker proteins C cluster of differentiation 63 (CD63) and C cluster of differentiation 9 (CD9) were expressed in the extracted exosomes, and exosome particle size was 80 to 150 nm; miR-22-3p was significantly lowly expressed in patients with premature ovarian failure and ovarian granulosa cells induced by CTX (P < 0.05), but was significantly highly expressed in ovarian granulosa cells incubated with exosomes derived from bone marrow mesenchymal stem cells (P < 0.05); the activity of granulosa cells in the CTX group was significantly lower than that in the control group, but was significantly higher in the CTX+miR-22-3p group or CTX+Exosomes group than that in the control group (P < 0.05); the apoptosis rate of granulosa cells in the CTX group was significantly higher than that in the control group, but was significantly lower in the CTX+miR-22-3p or CTX+Exosomes group than that in the control group (P < 0.05). Conclusion The miR-22-3p of extracellular vesicles derived from bone marrow mesenchymal stem cells can inhibit ovarian granulosa cell injury induced by CTX.

Objective To investigate the effect of different radiation doses on the accuracy of iodine quantification and CT value of virtual monochromatic image with a muti-spectral CT iodine phantom using second-generation dual-layer detector spectral CT(DLCT).Methods The second-generation DLCT scan abdominal phantom was used,which consisted of six different concentrations of iodine rods at 4,8 and 12 g/L.The scan parameters were set at 100,120 and 140 kVp tube voltages,and six radiation exposure sequences were scanned at each tube voltage:5,10,15,20,25 and 30 mGy.Measure the iodine concentration value and the CT value at the single level of 50,70 and 100 keV,and calculate the absolute error value of iodine concentration and CT value under different radiation exposure.SPSS 22.0 software single factor analysis of variance was used to compare the difference between the io-dine concentration and the absolute error of CT value under different Radiation exposure,and LSD method was used to compare the two groups.Results The absolute error of CT value of 4 g/L iodine rod with 10～30 mGy scanning at 100 keV energy level was significantly higher than that of 12 g/L and 8 g/L(P＜0.05),and the absolute error of iodine concentration measurement of other iodine rods with different concentrations at different Radiation exposure had no statistical difference.Conclusions The second-generation double-layer detector spectral CT has high meas-urement stability for iodine concentration quantification and CT value accuracy under conditions of radiation dose greater than 5 mGy.

Objective To investigate the influence from natural and constrained modal of the hip joint on internal fixation after implantation of hollow screws and locking plates for treating femoral neck fractures. Methods CT image data of a patient with femoral neck fracture were analyzed, boundary of the hip joint was extracted to generate a three-dimensional (3D) model of the hip joint, and the assembly of common internal fixation models and hip joint models was established. Finite element simulation was then conducted, with focus on vibration characteristics. Results Vibration had a certain effect on internal fixation stability of hollow screws and locking plates. The occurrence of torsion would destroy the fixation environment of hollow screws and locking plates, resulting in a small displacement at the fracture end. In a constrained state, the modal frequency range was lower. Deformation of the vibration shape mostly occurred at proximal end of the femoral head, leading to the loosening phenomenon of internal fixation and prosthesis. A method for judging the stability of internal fixation was proposed from the perspective of vibration characteristics, and it was found that the stability of internal fixation with hollow screws was better than that of proximal locking plates. Conclusions For choosing internal fixation, influences from natural frequency of the screw and plate should be considered, so as to avoid natural frequency of the femur, which may cause resonance. The results can be used as a guidance for the selection of internal fixation materials and the design of structure and configuration.

Extracellular vesicles (EVs) are membrane-bound particles actively released by cells. In prokaryotes and eukaryotes, EVs are effective bridges for communication between cells. EVs carry biological macromolecules, including proteins, lipids and nucleic acid, which affects different physiological functions of parent cells and recipient cells. Among them, the microRNA carried by EVs is the most reported and plays an important role in physiological function of organisms. During the development of follicles, only a few follicles can fully develop and ovulate, whereas most of them undergo atresia at different stages of development. In the whole process of follicular development, the changes at each stage and the regulation mechanism of follicular atresia are not completely understood. In this paper, we introduced the types, characteristics, isolation methods and uses of EVs, and emphasized how microRNA carried by EVs in follicular fluid regulated follicular atresia from the aspects of different cytokines and hormones. Additionally, the application prospect of microRNA carried by EVs in follicular fluid in reproductive regulation and reproductive disease diagnosis was discussed. This paper is significant for studying the regulation of follicular development and the effective utilization of oocytes.
Animals ; Extracellular Vesicles/metabolism* ; Female ; Follicular Atresia ; Follicular Fluid ; MicroRNAs/metabolism* ; Oocytes

Objective:To analyze the evolutionary characteristics of the approximate whole genome of rotavirus G2P[4] type 2020BJ strain.Methods:The rotavirus genome was amplified by reverse transcription-polymerase chain reaction (RT-PCR), the amplified products were sequenced, and the sequences were subjected to phylogenetic analysis and homology analysis.Results:The approximate full-length 11 segments of human rotavirus G2P[4] type 2020BJ strain were obtained. Sequence analysis showed that the strain was G2-P[4]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genotype (DS-1-Like); evolutionary analysis shows that it is closely related to strains in Japan, India, Bangladesh, Italy and other countries; there are differences in the amino acids of antigenic epitopes between the closely related strains.Conclusions:There are differences in the amino acids of the epitopes of VP7 and VP4 of the five G2P[4] rotavirus strains that are closely related to 2020BJ, which may lead to different epidemic characteristics, and rotavirus surveillance should be strengthened.

Objective:To implement an intervention with community-dwelling middle-aged and elderly people through a new science promotion approach, and to evaluate changes in the level of bone health awareness.Methods:From April 2017 to June 2017, 200 community-dwelling middle-aged and elderly people aged 50 years and over were randomly included.They were divided into the intervention group receiving a new science promotion intervention(n=100, with 50 males and 50 females)and the control group(n=100, with 50 males and 50 females). Baseline and follow-up assessments of bone health awareness were performed before and after the one-year intervention.A questionnaire survey on preferences for different types of science promotion articles in WeChat accounts was conducted.Differences in scores for intervention effects before and after intervention were compared between the two groups.Potential influencing factors for the scale score were examined by a multi-factor analysis.Results:After one year of intervention, scores for intervention effects were higher in the intervention group than in the control group( P<0.001). In the intervention group, scores were higher after intervention than before intervention( P<0.001). In the control group, scores after follow-up were slightly higher than those at baseline(29.4±11.4 vs.27.9±10.1, P<0.001). The increase in scores before and after intervention was greater in the intervention group than in the control group( P<0.001). Baseline score, group designation, history of drinking and diabetes impacted the scores in middle-aged and elderly women, while other factors did not.Most middle-aged and older people preferred texts illustrated with pictures, and older people were more receptive to videos. Conclusions:The new science promotion method is beneficial to bone health management, can improve bone health awareness in middle-aged and elderly people.

Objective:To explore the clinical efficacy of dual-kidney transplantation from infant donors to adult recipients.Methods:From December 2012 to November 2020 in Organ Transplant Center First Affiliated Hospital Sun Yat-sen University, rertrospective reviews were conducted for clinical data of 25 pairs of infant donors and adult recipients. The survival rates were calculated for both recipients and transplanted kidneys at Year 1/3/5 post-transplantation. And the postoperative recovery status and the postoperative incidence of adverse events of recipients were observed.Results:The survival rates of recipients were all 95.8% at Year 1/3/5 and those of transplanted kidney and dealth-cancelling transplanted kidney all 87.2%. One case died due to acute inferior-wall cardiac infarction while three others lost renal functions for vascular thrombosis, ureteral stenosis and urinary fistula. Except for loss of renal function and death, the postoperative estimated golmerular fitration rate was (99.35±21.78), (103.11±29.20) and (114.99±28.55) ml/(min·1.73 m 2) at Year 1/2/3 respectively. Conclusions:Selecting proper recipients, standardizing donor acquisition and surgical procedures and strengthening perioperative managements may expand the donor pool. The overall outcomes are excellent for adult recipients with dual-kidney transplantation from donations after infants' death.