In order to establish a highly efficient,convenient,and effective circular RNA(circRNA)vaccine preparation system,enhanced green fluorescent protein(EGFP)circRNA was constructed using permuted intron exon(PIE)strategy based on type Ⅰ introns.Then,circRNA circularization rates of RNA after in vitro transcription(IVT),primary circularization(IVC1),and secondary circularization(IVC2)were compared after purification.The constructed circRNA system was fur-ther applied to porcine reproductive and respiratory syndrome virus(PRRSV),and two circRNAs based on PRRSV GP5 protein were constructed and developed for in vitro expression.Results showed that circularization rates and protein expression effects of EGFP circRNA in IVC1 RNA and IVC2 RNA were similar,but both were significantly better than those of IVT RNA.Purity of EGFP circRNA reached 74％,and purities of two PRRSV GP5 protein circRNAs constructed using this preparation system were 71％and 64％,respectively.Western blot and indirect immunofluo-rescence assay(IFA)results indicated that both of the PRRSV GP5 protein circRNAs were suc-cessfully expressed.The results demonstrated that an easy-to-operate,low-cost circRNA prepara-tion system with high circularization rate was successfully constructed.Two PRRSV GP5 protein circRNA vaccines were successfully prepared using this system and expressed efficiently,which provides a reference for the development of animal circRNA vaccines and novel candidate vaccines against PRRSV.

Objective To evaluate the practice effect of medical record writing training based on standardized patients and explore its application in prosthodontics practice.Methods Seventy-one undergraduate interns were randomly divided into two groups.At the first week of clinical practice,the test group(n=35)adopted the SP-based medical record writing training,and after the training,the students'evaluation of the teaching effect was investigated by questionnaire.And the control group(n=36)received traditional lectured medical record writing training.One week later,the same medical record writing exam was performed in the two groups.The scores of medical record writing of different teaching model were compared,and the evaluation of the teaching effect in the test group was carried out.Results The exam score of medical record writing of the test group(88.80±4.60)was significantly higher than that of the control group(84.92±5.51),and the difference was statistically significant(P=0.002).88.57％of the students in the test group were satisfied with the SP-based medical record writing training.The satisfaction score was 8.94.Conclusion Medical record writing training is a long-term clinical practice.SP-based medical record writing training is helpful to improve the medical record writing quality of medical students.

Objective To investigate the value of gadolinium-ethoxybenzyl-diethylenetriaminepentaacetic acid(Gd-EOB-DTPA)enhanced MRI and T1 mapping for evaluating vessels encapsulating tumor clusters(VETC)caused by hepatocellular carcinoma(HCC).Methods Totally 112 cases of HCC confirmed by surgical pathology were retrospectively enrolled and categorized into positive group(n=46)and negative group(n=66)based on the presence or absence of VETC.Gd-EOB-DTPA enhanced MRI features and T1 mapping parameters were compared between groups.Multivariate logistic regression analysis was used to identify independent risk factors for HCC VETC,and a combined nomogram model was developed.Receiver operating characteristic(ROC)curve was plotted,and the area under the curve(AUC)was calculated to evaluate the efficacy of the identified independent risk factors and nomogram model for predicting HCC VETC.Results The maximum diameter of HCC,incidence of non-smooth tumor margin,prevalence of peritumoral hypo-intensity on hepatobiliary phase(HBP),as well as T1 value of HBP(T1post)in positive group were all higher,while the change rate of T1 value(ΔT1％)in positive group was lower than those in negative group(all P＜0.05).The increased maximum diameter,non-smooth margin,peritumoral hypo-intensity on HBP and low ΔT1％were all independent risk factors for HCC VETC(all P＜0.05),with AUC for predicting HCC VETC of 0.680,0.675,0.686 and 0.752,respectively.The nomogram model developed based on the combination of the above factors showed sensitivity of 78.79％,specificity of 71.74％and AUC of 0.839 for predicting HCC VETC.Conclusion Gd-EOB-DTPA enhanced MRI combined with T1 mapping could be used to effectively predict HCC VETC.

In order to establish a highly efficient,convenient,and effective circular RNA(circRNA)vaccine preparation system,enhanced green fluorescent protein(EGFP)circRNA was constructed using permuted intron exon(PIE)strategy based on type Ⅰ introns.Then,circRNA circularization rates of RNA after in vitro transcription(IVT),primary circularization(IVC1),and secondary circularization(IVC2)were compared after purification.The constructed circRNA system was fur-ther applied to porcine reproductive and respiratory syndrome virus(PRRSV),and two circRNAs based on PRRSV GP5 protein were constructed and developed for in vitro expression.Results showed that circularization rates and protein expression effects of EGFP circRNA in IVC1 RNA and IVC2 RNA were similar,but both were significantly better than those of IVT RNA.Purity of EGFP circRNA reached 74％,and purities of two PRRSV GP5 protein circRNAs constructed using this preparation system were 71％and 64％,respectively.Western blot and indirect immunofluo-rescence assay(IFA)results indicated that both of the PRRSV GP5 protein circRNAs were suc-cessfully expressed.The results demonstrated that an easy-to-operate,low-cost circRNA prepara-tion system with high circularization rate was successfully constructed.Two PRRSV GP5 protein circRNA vaccines were successfully prepared using this system and expressed efficiently,which provides a reference for the development of animal circRNA vaccines and novel candidate vaccines against PRRSV.

Objective To evaluate the practice effect of medical record writing training based on standardized patients and explore its application in prosthodontics practice.Methods Seventy-one undergraduate interns were randomly divided into two groups.At the first week of clinical practice,the test group(n=35)adopted the SP-based medical record writing training,and after the training,the students'evaluation of the teaching effect was investigated by questionnaire.And the control group(n=36)received traditional lectured medical record writing training.One week later,the same medical record writing exam was performed in the two groups.The scores of medical record writing of different teaching model were compared,and the evaluation of the teaching effect in the test group was carried out.Results The exam score of medical record writing of the test group(88.80±4.60)was significantly higher than that of the control group(84.92±5.51),and the difference was statistically significant(P=0.002).88.57％of the students in the test group were satisfied with the SP-based medical record writing training.The satisfaction score was 8.94.Conclusion Medical record writing training is a long-term clinical practice.SP-based medical record writing training is helpful to improve the medical record writing quality of medical students.

Objective To investigate the value of gadolinium-ethoxybenzyl-diethylenetriaminepentaacetic acid(Gd-EOB-DTPA)enhanced MRI and T1 mapping for evaluating vessels encapsulating tumor clusters(VETC)caused by hepatocellular carcinoma(HCC).Methods Totally 112 cases of HCC confirmed by surgical pathology were retrospectively enrolled and categorized into positive group(n=46)and negative group(n=66)based on the presence or absence of VETC.Gd-EOB-DTPA enhanced MRI features and T1 mapping parameters were compared between groups.Multivariate logistic regression analysis was used to identify independent risk factors for HCC VETC,and a combined nomogram model was developed.Receiver operating characteristic(ROC)curve was plotted,and the area under the curve(AUC)was calculated to evaluate the efficacy of the identified independent risk factors and nomogram model for predicting HCC VETC.Results The maximum diameter of HCC,incidence of non-smooth tumor margin,prevalence of peritumoral hypo-intensity on hepatobiliary phase(HBP),as well as T1 value of HBP(T1post)in positive group were all higher,while the change rate of T1 value(ΔT1％)in positive group was lower than those in negative group(all P＜0.05).The increased maximum diameter,non-smooth margin,peritumoral hypo-intensity on HBP and low ΔT1％were all independent risk factors for HCC VETC(all P＜0.05),with AUC for predicting HCC VETC of 0.680,0.675,0.686 and 0.752,respectively.The nomogram model developed based on the combination of the above factors showed sensitivity of 78.79％,specificity of 71.74％and AUC of 0.839 for predicting HCC VETC.Conclusion Gd-EOB-DTPA enhanced MRI combined with T1 mapping could be used to effectively predict HCC VETC.

Objective:To explore the independent risk factors for the occurrence and aggravation of lower back pain (LBP) in patients with Parkinson′s disease (PD), in order to provide reference for clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the case data of 309 PD patients who visited the Affiliated Yixing Hospital of Jiangsu University from June 2018 to May 2020. The KING Parkinson′s Disease Pain Scale (KPPS) was used to quantitatively evaluate the LBP of PD patients, who were divided into LBP group and Non LBP group. The general clinical data, PD related data, and imaging data of the two groups were compared and analyzed. Binary logistic regression analysis was used to evaluate independent risk factors for LBP in PD patients. Pearson correlation analysis was conducted between KPPS scores and various factors, and linear regression analysis was used to identify the relevant risk factors that exacerbate LBP in PD patients.Results:Compared with the Non LBP group, the LBP group had lower bone mineral density (BMD) and a lower proportion of patients who engaged in daily exercise. The difference between the two groups was statistically significant (all P<0.05). Compared with the Non LBP group, patients in the LBP group had a longer course of illness, higher stiffness scores, a higher proportion of patients with fluctuating symptoms, higher UPDRS-Ⅲ scores, and a higher proportion of patients with thoracolumbar fascial injury (TLFI) and lumbar sagittal imbalance. The differences between the two groups were statistically significant (all P<0.05). The results of binary logistic regression analysis showed that combined TLFI ( OR=2.773, 95% CI: 1.219-6.309, P=0.015), combined lumbar sagittal imbalance ( OR=4.835, 95% CI: 2.244-10.421, P<0.001), and lower BMD ( OR=2.818, 95% CI: 1.767-4.493, P<0.001) were risk factors for LBP in PD patients. The KPPS score was correlated with BMD and TLFI ( r=-0.146, 0.294, all P<0.05). The linear regression results showed that the merged TLFI ( B=2.271, β=0.285, P<0.001) was positively correlated with KPPS score, indicating a risk factor. Conclusions:The combination of TLFI, lumbar sagittal imbalance, and lower BMD is closely related to the occurrence of LBP in PD patients, and the combination of TLFI is an independent risk factor for exacerbating LBP symptoms. Clinical attention should be paid to the prevention and treatment of TLFI in PD patients.

The five movements and six qi theory(hereinafter referred to as"movement qi theory")runs through Liu Wansu's med-ical theoretical system,which is mainly reflected in its influence on the theory of pathogenesis,the creation of fiery heat theory,and the influence of Liu Wansu's clinical diagnosis and treatment thoughts.The influence of movement qi theory on Liu Wansu's pathogen-esis theory includes three aspects:six qi alternation on pathogenesis,five movement hyperactivities on pathogenesis,and the root and branch of movement qi on pathogenesis.Liu's understanding of the pathogenesis of six qi and the philosophical thought of"water is good and fire is evil"mainly influenced the creation of Liu Wansu's"fire theory".The influence of movement qi theory on Liu Wansu's clinical diagnosis and treatment thoughts is mainly reflected in the classification of viscera,meridian and collateral diseases based on the theory of root and branch of middle qi,the diagnosis of pulses based on movement qi theory,the deduction of disease transmission and change using movement qi thinking,and the preparation of medicines based on qi.Liu Wansu initiated the innovation and debate of medical theory in the Jin and Yuan Dynasties,and had a profound impact on the rise of warm febrile diseases in the Ming and Qing Dynasties.

Objective: To explore the effects of grandparent involvement in parenting on health related behaviors among urban preschool children, so as to provide a basis for the development of interventions for unhealthy behaviors among preschool children with different parenting pattern. Methods: A total of 5 431 preschool children aged 3-6 years old in Xinbei District, Changzhou City, were selected by census method during January to October, 2022. General household characteristics and health related behaviors of children were investigated by questionnaires, and the differences in health related behaviors with different parenting pattern involving grandparents were explored by the χ 2 test. A multifactorial Logistic regression model was used to analyze the effects of grandparenting on preschool children s health behaviors. Results: About 51.2% of preschoolers were raised by both parents, 46.1 % were co parented by grandparents, and 2.7% were raised by grandparents. After multifactorial Logistic regression analysis controlling for age, gender, annual household income, and family structure, children co parented by grandparents had a reduced risk of screen exposure time>1 h/d ( OR =0.79, P <0.05), with increased risks of outdoor activity time<2 h/d ( OR =1.30, P < 0.05 ), sleep duration <10 h/d ( OR =1.31, P <0.05), sleep difficulties ( OR =1.39, P <0.05) and adverse eating behaviors ( OR =1.20， P < 0.05), compared with children parented by both parents. Children raised by grandparents were at increased risk for outdoor activity time<2 h/d ( OR =2.19) and unhealthy eating behaviors ( OR =1.39) ( P <0.05). Conclusions Compared with children raised by both parents, the incidence of unhealthy behaviors is increased in children raised by parents and grandparents and in children raised by full grandparents. Active intervention should be focused on unhealthy behaviors of preschool children with pareting involving grandparents to promote the physical and mental health of preschool children.

Objective:To evaluate the quality of the published breast cancer screening guidelines to provide a reference for domestic studies in the future.Methods:PubMed, Embase, Cochrane Library, Web of Science, SinoMed, China National Knowledge Infrastructure, VIP, and Wanfang Data were searched to identify breast cancer screening guidelines on until August 2020. Two reviewers screened literature and extracted data independently. The Appraisal of Guidelines for Research & Evaluation Ⅱ(AGREEⅡ) and Reporting Items for Practice Guidelines in Healthcare(RIGHT) tools were used to evaluate the quality of the included guidelines.Results:A total of 15 breast cancer screening guidelines were included, of which seven were published in the United States, with publication years focusing on 2015 to 2019, and 11 guidelines had updated versions. "Rigour of development" (47.0%±22.1%) and "Applicability" (44.0%±15.1%) of AGREEⅡ scored lower than other domains. "Review and quality assurance" (46.7%±39.9%) and "Funding, declaration, and management of interests" (41.7%±24.4%) of RIGHT were reported poorer than others. There were six guidelines recommended and another nine recommended with modifications based on the overall AGREEⅡ score. There were four guidelines with a good level, and another 11 were with a moderate level of RIGHT. The National Comprehensive Cancer Network published the best overall quality guidelines in 2018 (AGREEⅡ: 83.3%, RIGHT: 80.0%) and by the American Cancer Society in 2015 (AGREEⅡ: 83.3%, RIGHT: 85.7%).Conclusion:The quality of breast cancer screening guidelines was predominantly of moderate quality, and greater attention should be paid to the guideline development process and quality control of the guidelines.