The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

Objective: To investigate the willingness to accept preventive treatments and its related factors among college freshmen with latent tuberculosis infection (LTBI), so as to provide the evidence for preventive treatment intervention measures for students with LTBI. Methods: Cluster sampling method was used to select 368 LTBI freshmen from 8 colleges and universities in Changzhou in September 2023, who conducted a questionnaire survey on the willingness to receive preventive treatment. General demographic data were collected and relevant data were collected using tuberculosis knowledge scale, General Anxiety Disorder-7 (GAD-7), Patient Health Questionnaire-9 (PHQ-9), Adaptation, Partnership, Growth, Affection and Resolve (APGAR), and a self developed Stigma Scale. A binary Logistic regression model was constructed with the willingness to accept preventive treatment as the dependent variable to analyze the willingness to accept preventive treatment and the influencing factors. Results: A total of 253 LTBI college freshmen were willing to take preventive treatment, the acceptance rate was 68.75%. The rate of willingness to accept preventive treatment for LTBI was higher among students whose fathers had an education level of high school, compared to those whose fathers had an education level of junior high school or below ( OR =2.16, P <0.05). LTBI students whose per capita family income was >5 000-10 000 yuan and >10 000 yuan were more willing to accept LTBI preventive treatment than those whose per capita family income was <3 000 yuan ( OR =2.72, 4.46, P <0.05). LTBI students who engaged in physical exercise for more than 2 hours per week were more willing to accept than those who exercised less than 0.5 hours per week ( OR =1.91, P <0.05). LTBI students with high levels of tuberculosis knowledge and stigma were more likely to receive preventive treatment ( OR =1.18, 1.11, P < 0.05). LTBI students with high PHQ-9 ( OR =0.85) and GAD-7 ( OR =0.92) scores were more likely to refuse preventive treatment ( P <0.05). Conclusion The present study revealed a moderate level of willingness of LTBI students to preventive treatment in Changzhou City, and the acceptance is affected by family factors, healthy lifestyles, tuberculosis knowledge and psychological status.

BACKGROUND:In recent years,with the development of biological scaffold materials and bioprinting technology,tissue-engineered bone has become a research hotspot in bone defect repair. OBJECTIVE:To summarize the current treatment methods for bone defects,summarize the biomaterials and bioprinting technology for preparing tissue-engineered bone scaffolds,and explore the application of biomaterials and printing technology in tissue engineering and the current challenges. METHODS:Search terms were"bone defect,tissue engineering,biomaterials,3D printing technology,4D printing technology,bioprinting,biological scaffold,bone repair"in Chinese and English.Relevant documents published from January 1,2009 to December 1,2022 were retrieved on CNKI,PubMed and Web of Science databases.After being screened by the first author,high-quality references were added.A total of 93 articles were included for review. RESULTS AND CONCLUSION:The main treatment methods for bone defects include bone transplantation,membrane-guided regeneration,gene therapy,bone tissue engineering,etc.The best treatment method is still uncertain.Bone tissue engineering technology is a new technology for the treatment of bone defects.It has become the focus of current research by constructing three-dimensional structures that can promote the proliferation and differentiation of osteoblasts and enhance the ability of bone formation.Biological scaffold materials are diverse,with their characteristics,advantages and disadvantages.A single biological material cannot meet the demand for tissue-engineered bone for the scaffold.Usually,multiple materials are combined to complement each other,which is to meet the demand for mechanical properties while taking into account the biological properties of the scaffold.Bioprinting technology can adjust the pore of the scaffold,build a complex spatial structure,and is more conducive to cell adhesion,proliferation and differentiation.The emerging 4D printing technology introduces"time"as the fourth dimension to make the prepared scaffold dynamic.With the synchronous development of smart materials,4D printing technology provides the possibility of efficient repair of bone defects in the future.

Objective:To evaluate the knowledge of autism among child health care professionals in primary health care institutions.Methods:The study was a cross-sectional survey. An online questionnaire survey was conducted from February to March 2023 in primary health care institutions in Guangzhou to investigate the knowledge on autism among medical staff engaged in children′s health services and the influencing factors.Results:A total of 341 questionnaires were returned and 312 questionnaires were valid with a recovery rate of 91.5%. The age of 312 respondents was (35.9±7.9) years, of which 303 (97.1%) were female. One hundred and fifty-two (48.7%) child health care professionals in primary health care institutions had received specialist training in assessing the psychological and behavioral development of children, and only 139 (44.6%) reported that they were aware of the"five no"principle for early identification of autism. The questionnaire scores were 88.1% pass (275/312) and 53.2% excellent (166/312). The three questions with low accuracy were: autism can be cured with drugs, autism has a genetic basis and rehabilitation training has no effect, and the accuracy for these questions was 42.6% (133/312), 52.2% (163/312) and 70.2% (219/312), respectively. The passing of autism-related knowledge was positively associated with receiving relevant training ( OR=2.585, 95% CI:1.200-5.569), and the excellence was positively associated with the highest education ( OR=1.939, 95% CI:1.220-3.083) and receiving relevant training ( OR=2.016, 95% CI:1.247-3.260). Conclusions:There is a need for more professional training in autism knowledge among child health care professionals in primary health care institutions.

Objective To observe the value of angle of progression(AOP)measured with transperineal ultrasound in the second stage of labor for predicting delivery mode using meta-analysis.Methods Articles published in Chinese or English focused on the prediction of delivery mode using AOP measured with ultrasound during the second stage of labor were searched in databases from January 1,2018 to August 31,2023.The literature were screened by 2 researchers according to the inclusion and exclusion criteria,and information were extracted to assess the value of AOP measured in the second stage of labor with transperineal ultrasound for predicting delivery mode.The quality of the included literature were evaluated using Quality Evaluation Tool for Diagnostic Tests-2(QUADAS-2).Results Eleven studies including 2 315 parturient women were enrolled.Meta-analysis showed that the combined sensitivity(Sen),combined specificity(Spe),combined positive likelihood ratio(+LR),combined negative likelihood ratio(-LR),and combined diagnostic ratio(DOR)of AOP measured with transperineal ultrasound in the second stage of labor for predicting delivery mode was 0.80[95％CI(0.76,0.83)],0.83[95％CI(0.78,0.88)],4.8[95％CI(3.5,6.6)],0.24[95％CI(0.20,0.29)]and 20[95％CI(13,30)],respectively,while area under the summary receiver operating characteristic curve of 0.88.Conclusion AOP measured with transperineal ultrasound in the second stage of labor had fine efficiency for predicting delivery mode,which could be regarded as a useful predictor for probability of natural delivery.

Objective:To evaluate the hemodynamic changes and short-term (one year) outcomes after superficial temporal artery(STA)-middle cerebral artery (MCA) bypass by vascular ultrasonography.Methods:Operation group included a total of 41 hemispheres of 38 patients who underwent STA-MCA bypass for severe stenosis or occlusion of MCA or severe stenosis or occlusion of internal carotid artery(ICA), or Moyamoya disease at Xuanwu Hospital between August 2017 and June 2020. The following examinations were performed in all patients: cranial MRI, CT perfusion imaging, and cerebral angiography. All of the patients were retrospectively followed up for one year postsurgery. Normal group included a total of 40 hemispheres of 20 healthy people who had been examined by vascular ultrasonography of carotid artery and intracranial artery. Inner diameter, peak systolic velocity (PSV), end-diastolic velocity (EDV), mean velocity (MV), resistance index (RI), pulsatility index (PI) and flow of STA, PSV, EDV, MV, PI, RI of external carotid artery (ECA) and PSV, EDV, MV, PI, RI of MCA at 1 week, 6 months and 12 months after STA-MCA bypass of operation group were compared with normal group.Results:①Inner diameter, PSV, MV and flow of STA were increased significantly in operation group at 1 week, 6 and 12 months than normal group (all P<0.05). The flow of STA was decreased significantly from 1 week to 12 months after operation (all P<0.05), but Inner diameter, PSV and MV were only decreased gradually from 1 week to 12 months after operation (all P>0.05). ②STA/ECA PI and RI decreased significantly after operation compared with normal group (all P<0.01). Conclusions:Vascular ultrasonography is a non-invasive examination which can objectively evaluate the extracranial and intracranial hemodynamic changes after STA-MCA bypass, and provide reference effectiveness of the operation. The flow of STA has decreased gradually from 1 week to 12 months after operation to achieve the homeostasis, but it is still higher than normal.STA/ECA PI and RI can be a steady indirect pointer to show the fluency of bypass.

Humans ; Pulmonary Disease, Chronic Obstructive ; Smoke/adverse effects* ; Tobacco ; Tobacco Smoke Pollution

Objective:To investigate the causes of obsolete uterine perforation with tubal incarceration and summarize the clinical manifestations, differential diagnosis and auxiliary examination signs of tubal incarceration.Methods:Two cases of obsolete uterine perforation with tubal incarceration were retrospectively analyzed and the literatures were reviewed.Results:Oviduct incarceration was observed during operation, and postoperative pathological examination confirmed that the incarcerated tissue was oviduct.Conclusion:Obsolete uterine perforation complicated with fallopian tube incarceration was lack of specific manifestations, and the patients undergoing intrauterine manipulation should be given comprehensive examination for early diagnosis and treatment to improve prognosis of patients.

Objective:To investigate the causes of obsolete uterine perforation with tubal incarceration and summarize the clinical manifestations, differential diagnosis and auxiliary examination signs of tubal incarceration.Methods:Two cases of obsolete uterine perforation with tubal incarceration were retrospectively analyzed and the literatures were reviewed.Results:Oviduct incarceration was observed during operation, and postoperative pathological examination confirmed that the incarcerated tissue was oviduct.Conclusion:Obsolete uterine perforation complicated with fallopian tube incarceration was lack of specific manifestations, and the patients undergoing intrauterine manipulation should be given comprehensive examination for early diagnosis and treatment to improve prognosis of patients.