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 efficacy and safety of domestic RECO flow restoration device in endovascular treatment of acute ischemic stroke(AIS)patients in a real-world setting.Methods From January 2019 to June 2021,consecutive patients with acute intracranial vessel occlusion treated with RECO device from 8 stroke centers in Jiangsu Province were analyzed retrospectively.Baseline and clinical data were collected,including age,gender,baseline modified Rankin scale(mRS)scores,National Institutes of Health stroke scale(NIHSS)score on admission,trial of Org 10172 in acute stroke treatment(TOAST)classification,responsible location of occlusion,wake-up stroke,and medical history(hypertension,diabetes mellitus,hyperlipidemia,atrial fibrillation,coronary heart disease,peripheral arterial disease),smoking,and alcohol consumption.Furthermore,surgical parameters were collected,including time indicators(intervals between symptom onset,admission,puncture and recanalization),preoperative intravenous thrombolysis,general anesthesia,combination of aspiration,number of passes,first pass effect(modified thrombolysis in cerebral infarction[mTICI]grade 3 achieved with s single stent-based thrombectomy),rescue therapy(including thrombectomy with other stents,intra-arterial urokinase thrombolysis,intra-arterial application of tirofiban,emergency angioplasty[stent implantation,balloon dilatation,balloon dilatation combined with stent implantation]),balloon guided catheter and NIHSS score at discharge.The effectiveness indicators included immediate successful recanalization after the operation(mTICI grade ≥ 2b),complete recanalization(mTICI grade 3),and a good prognosis at 90 d after the operation(mRS score ≤2).The safety indicators included symptomatic intracranial hemorrhage(sICH)within 24 h after the operation,device-related intraoperative complications(such as thrombus escape,iatrogenic dissection,bleeding at the surgical site),and all-cause mortality within 90 days after the operation.All patients were divided into the anterior circulation occlusion group and the posterior circulation occlusion group according to the location of the responsible occluded vessel,and the effectiveness and safety indexes were analyzed.Results A total of 366 patients with AIS caused by intracranial vessel occlusion who underwent mechanical thrombectomy with the domestic RECO thrombectomy stent were included.The average age was(69±11)years,with 54 patients(14.8％)over 80 years old and 221 patients(60.4％)male.There were 296 patients in the anterior circulation occlusion group and 70 patients in the posterior circulation occlusion group.(1)Immediate successful recanalization(mTICI grade≥2b)was achieved in 337 patients(92.1％),and complete recanalization was achieved in 282 patients(77.0％)immediately after the operation.The average number of thrombectomies was(1.9±1.0)times,and 141 patients(38.5％)achieved recanalization on the first attempt.9 patients(2.5％)used other stents during the operation,and 65 patients(17.8％)underwent emergency angioplasty.The incidence of sICH within 24 h after the operation was 10.9％(40/366),the good prognosis rate at 90 d after the operation was 53.6％(196/366),and the all-cause mortality rate at 90 d after the operation was 23.0％(84/366).The incidence of device-related intraoperative complications was 3.6％(13/366).The median time from arterial puncture to recanalization was 75(52,110)min,and the median time from onset to recanalization was 370(280,488)min.(2)The good prognosis rates at 90 d after the operation in the anterior and posterior circulation occlusion groups were 55.1％(163/296)and 47.1％(33/70),respectively.The immediate successful recanalization rates were 93.2％(276/296)and 87.1％(61/70),respectively.The immediate complete recanalization rates were 79.1％(234/296)and 68.6％(48/70),respectively.The incidence of sICH within 24h after the operation was 12.2％(36/296)and 5.7％(4/70),respectively.The incidence of device-related intraoperative complications was 3.7％(11/296)and 2.9％(2/70),respectively.The all-cause mortality rate within 90 d after the operation was 21.6％(64/296)and 28.6％(20/70),respectively.There were no statistically significant differences between the two groups in the above aspects(all P＞0.05).Conclusions The application of the domestic RECO thrombectomy stent in the treatment of AIS caused by intracranial vessel occlusion can effectively and safely achieve vascular recanalization.The results of this study still need to be further verified by prospective controlled studies.

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 efficacy and safety of domestic RECO flow restoration device in endovascular treatment of acute ischemic stroke(AIS)patients in a real-world setting.Methods From January 2019 to June 2021,consecutive patients with acute intracranial vessel occlusion treated with RECO device from 8 stroke centers in Jiangsu Province were analyzed retrospectively.Baseline and clinical data were collected,including age,gender,baseline modified Rankin scale(mRS)scores,National Institutes of Health stroke scale(NIHSS)score on admission,trial of Org 10172 in acute stroke treatment(TOAST)classification,responsible location of occlusion,wake-up stroke,and medical history(hypertension,diabetes mellitus,hyperlipidemia,atrial fibrillation,coronary heart disease,peripheral arterial disease),smoking,and alcohol consumption.Furthermore,surgical parameters were collected,including time indicators(intervals between symptom onset,admission,puncture and recanalization),preoperative intravenous thrombolysis,general anesthesia,combination of aspiration,number of passes,first pass effect(modified thrombolysis in cerebral infarction[mTICI]grade 3 achieved with s single stent-based thrombectomy),rescue therapy(including thrombectomy with other stents,intra-arterial urokinase thrombolysis,intra-arterial application of tirofiban,emergency angioplasty[stent implantation,balloon dilatation,balloon dilatation combined with stent implantation]),balloon guided catheter and NIHSS score at discharge.The effectiveness indicators included immediate successful recanalization after the operation(mTICI grade ≥ 2b),complete recanalization(mTICI grade 3),and a good prognosis at 90 d after the operation(mRS score ≤2).The safety indicators included symptomatic intracranial hemorrhage(sICH)within 24 h after the operation,device-related intraoperative complications(such as thrombus escape,iatrogenic dissection,bleeding at the surgical site),and all-cause mortality within 90 days after the operation.All patients were divided into the anterior circulation occlusion group and the posterior circulation occlusion group according to the location of the responsible occluded vessel,and the effectiveness and safety indexes were analyzed.Results A total of 366 patients with AIS caused by intracranial vessel occlusion who underwent mechanical thrombectomy with the domestic RECO thrombectomy stent were included.The average age was(69±11)years,with 54 patients(14.8％)over 80 years old and 221 patients(60.4％)male.There were 296 patients in the anterior circulation occlusion group and 70 patients in the posterior circulation occlusion group.(1)Immediate successful recanalization(mTICI grade≥2b)was achieved in 337 patients(92.1％),and complete recanalization was achieved in 282 patients(77.0％)immediately after the operation.The average number of thrombectomies was(1.9±1.0)times,and 141 patients(38.5％)achieved recanalization on the first attempt.9 patients(2.5％)used other stents during the operation,and 65 patients(17.8％)underwent emergency angioplasty.The incidence of sICH within 24 h after the operation was 10.9％(40/366),the good prognosis rate at 90 d after the operation was 53.6％(196/366),and the all-cause mortality rate at 90 d after the operation was 23.0％(84/366).The incidence of device-related intraoperative complications was 3.6％(13/366).The median time from arterial puncture to recanalization was 75(52,110)min,and the median time from onset to recanalization was 370(280,488)min.(2)The good prognosis rates at 90 d after the operation in the anterior and posterior circulation occlusion groups were 55.1％(163/296)and 47.1％(33/70),respectively.The immediate successful recanalization rates were 93.2％(276/296)and 87.1％(61/70),respectively.The immediate complete recanalization rates were 79.1％(234/296)and 68.6％(48/70),respectively.The incidence of sICH within 24h after the operation was 12.2％(36/296)and 5.7％(4/70),respectively.The incidence of device-related intraoperative complications was 3.7％(11/296)and 2.9％(2/70),respectively.The all-cause mortality rate within 90 d after the operation was 21.6％(64/296)and 28.6％(20/70),respectively.There were no statistically significant differences between the two groups in the above aspects(all P＞0.05).Conclusions The application of the domestic RECO thrombectomy stent in the treatment of AIS caused by intracranial vessel occlusion can effectively and safely achieve vascular recanalization.The results of this study still need to be further verified by prospective controlled studies.

Objective:To explore the prevalence and influencing factors of allergic conjunctivitis in Baotou area at different times.Methods:A cross-sectional study was conducted in January 2021, 111 patients with allergic conjunctivitis who were admitted to the Department of Ophthalmology, the Second Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology in 2020 were selected as the research subjects. The basic information, allergic conditions, living habits and other information of the patients were obtained by self-made questionnaire. The pollen concentration and allergy related indicators(allergic duration and frequency, eye itching score, eye burn score, eye foreign body sensation score, tears score and the total scores) of 2019 and 2020 were analyzed by paired sample t-test. The pollen concentration of 2019 and 2020 were analyzed by Wilcoxon signed rank test. The allergy related indicators of different genders were analyzed by independent sample t-test. The allergy related indicators of different ages were analyzed by one-way analysis of variance. The relationship between the length of wearing masks and the allergy related indicators were analyzed by Spearman correlation analysis. Results:Among the 111 patients with allergic conjunctivitis, 54 were males and 57 were females, ranging in age from 8 to 69 years old, with a median age of 35 years old and an average age of (36.77±13.18) years old. The average pollen concentration in Baotou in 2019 was (125.35±222.64) grains/1 000 square millimeter, and the average pollen concentration in Baotou in 2020 was (107.38±137.29) grains/1 000 square millimeter. There was no significant difference in pollen concentration between the two years( Z=-0.178, P=0.859). The severity of allergic indicators in all patients in 2020 is significantly lower than in 2019( t values were 4.701,3.587,2.582,3.661,4.444,2.784,2.555,3.886,respectively, with P<0.05). The severity of allergic indicators in male patients is lower than that of female patients ( t values were -1.558, -1.257, -3.41, -3.085, -2.335, -2.897, -2.652, -4.124, respectively). The prevalence of allergic conjunctivitis dose not vary significant with age ( P values were 0.504, 0.095, 0.499, 0.265, 0.284, 0.655, 0.421, 0.976, respectively). In 2020, the average time that patients wear masks is (6.55±3.28) h/d, and there is a correlation with the difference in allergy duration ( r=0.191, P=0.045). Conclusion:The severity of allergic conjunctivitis in Baotou in 2020 was significantly improved compared with that in 2019, which may be related to masks wearing.

Objective:To explore the prevalence and influencing factors of allergic conjunctivitis in Baotou area at different times.Methods:A cross-sectional study was conducted in January 2021, 111 patients with allergic conjunctivitis who were admitted to the Department of Ophthalmology, the Second Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology in 2020 were selected as the research subjects. The basic information, allergic conditions, living habits and other information of the patients were obtained by self-made questionnaire. The pollen concentration and allergy related indicators(allergic duration and frequency, eye itching score, eye burn score, eye foreign body sensation score, tears score and the total scores) of 2019 and 2020 were analyzed by paired sample t-test. The pollen concentration of 2019 and 2020 were analyzed by Wilcoxon signed rank test. The allergy related indicators of different genders were analyzed by independent sample t-test. The allergy related indicators of different ages were analyzed by one-way analysis of variance. The relationship between the length of wearing masks and the allergy related indicators were analyzed by Spearman correlation analysis. Results:Among the 111 patients with allergic conjunctivitis, 54 were males and 57 were females, ranging in age from 8 to 69 years old, with a median age of 35 years old and an average age of (36.77±13.18) years old. The average pollen concentration in Baotou in 2019 was (125.35±222.64) grains/1 000 square millimeter, and the average pollen concentration in Baotou in 2020 was (107.38±137.29) grains/1 000 square millimeter. There was no significant difference in pollen concentration between the two years( Z=-0.178, P=0.859). The severity of allergic indicators in all patients in 2020 is significantly lower than in 2019( t values were 4.701,3.587,2.582,3.661,4.444,2.784,2.555,3.886,respectively, with P<0.05). The severity of allergic indicators in male patients is lower than that of female patients ( t values were -1.558, -1.257, -3.41, -3.085, -2.335, -2.897, -2.652, -4.124, respectively). The prevalence of allergic conjunctivitis dose not vary significant with age ( P values were 0.504, 0.095, 0.499, 0.265, 0.284, 0.655, 0.421, 0.976, respectively). In 2020, the average time that patients wear masks is (6.55±3.28) h/d, and there is a correlation with the difference in allergy duration ( r=0.191, P=0.045). Conclusion:The severity of allergic conjunctivitis in Baotou in 2020 was significantly improved compared with that in 2019, which may be related to masks wearing.