Objective To develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P＜0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P＜0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P＜0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation.

Objective:To investigate application of the teaching method of problem-based learning(PBL)combined with neuronavigation technique in neurosurgical teaching,so as to improve further the teaching quality.Methods:The teaching method of PBL combined with neuronavigation technique adopted the same PBL teaching approach with conventional teaching method,and the neuronavigation technique was applied to reconstruct three dimensional(3D)graphics for patients'imaging documents.The neuronavigation technique was used to display intraoperative operation process for students,and guide their operation in surgery,and conduct teaching for them.A total of 30 medical students who conducted internship at department of neurosurgery of Beijing Tiantan Hospital,Capital Medical University during January and December 2023 were selected.A random-number table method was adopted to divide them into conventional PBL teaching method group(n=15)and PBL combined with neuronavigation technique teaching method group(n=15).The students'assessment results between two teaching methods were compared when they completed internship.A self-made survey questionnaire was adopted to investigate students'satisfaction for two kinds of teaching methods.Results:The assessment results of theory,technical ability and imaging of PBL combined with neuronavigation technique teaching method group were respectively(90.27±2.66),(90.73±2.71)and(90.13±2.77),all of which were higher than those of conventional PBL teaching method,and the differences were significant(t=13.82,7.95,13.93,P<0.05).The satisfactions about case,teacher performance,auxiliary teaching tools,learning initiative,learning efficiency,the mastery of theoretical,anatomical and imaging knowledge,the mastery of operational skills,and communication and interaction in PBL combined with neuronavigation technique teaching method group were all higher than those of conventional PBL teaching method,and the differences were significant(t=10.96,5.98,6.51,4.64,3.83,4.80,6.95,7.54,9.47,5.93,P<0.05).Conclusion:The application of the teaching method that PBL combined with neuronavigation technique in the teaching of neurosurgery for medical students can improve the teaching quality of neurosurgery,and enhance the satisfaction of students for teaching method.

Objective:To investigate application of the teaching method of problem-based learning(PBL)combined with neuronavigation technique in neurosurgical teaching,so as to improve further the teaching quality.Methods:The teaching method of PBL combined with neuronavigation technique adopted the same PBL teaching approach with conventional teaching method,and the neuronavigation technique was applied to reconstruct three dimensional(3D)graphics for patients'imaging documents.The neuronavigation technique was used to display intraoperative operation process for students,and guide their operation in surgery,and conduct teaching for them.A total of 30 medical students who conducted internship at department of neurosurgery of Beijing Tiantan Hospital,Capital Medical University during January and December 2023 were selected.A random-number table method was adopted to divide them into conventional PBL teaching method group(n=15)and PBL combined with neuronavigation technique teaching method group(n=15).The students'assessment results between two teaching methods were compared when they completed internship.A self-made survey questionnaire was adopted to investigate students'satisfaction for two kinds of teaching methods.Results:The assessment results of theory,technical ability and imaging of PBL combined with neuronavigation technique teaching method group were respectively(90.27±2.66),(90.73±2.71)and(90.13±2.77),all of which were higher than those of conventional PBL teaching method,and the differences were significant(t=13.82,7.95,13.93,P<0.05).The satisfactions about case,teacher performance,auxiliary teaching tools,learning initiative,learning efficiency,the mastery of theoretical,anatomical and imaging knowledge,the mastery of operational skills,and communication and interaction in PBL combined with neuronavigation technique teaching method group were all higher than those of conventional PBL teaching method,and the differences were significant(t=10.96,5.98,6.51,4.64,3.83,4.80,6.95,7.54,9.47,5.93,P<0.05).Conclusion:The application of the teaching method that PBL combined with neuronavigation technique in the teaching of neurosurgery for medical students can improve the teaching quality of neurosurgery,and enhance the satisfaction of students for teaching method.

Background: Fecal microbiota transplantation (FMT) is a therapeutic approach targeting at intestinal microorganisms and is used in recent years for the treatment of Crohn's disease (CD), however, its efficacy and safety remain unclear. Aims: To systematically evaluated the efficacy and safety of FMT in CD patients. Methods: Studies of FMT in the treatment of CD were retrieved from Embase, Medline (via PubMed), Cochrane Library, Web of Science, CNKI, Wanfang Data, VIP, CBM from their inception to May 1, 2020. Literatures were independently screened by two reviewers, data were extracted and the quality of included studies was assessed by MINORS. Meta-analysis was conducted by RevMan 5.3 software. Results: Seven studies involving 256 CD patients were enrolled. Meta-analysis showed that total response rate of CD patients treated with FMT was 72% (95% CI: 0.66-0.77), while remission rate was 60% (95% CI: 0.51-0.69). The common adverse reactions were abdominal pain, diarrhea, abdominal bloat; no severe adverse reactions were found. Conclusions: The efficacy and safety of FMT for the treatment of CD are quite high as indicated in this study, however, high-quality randomized controlled trials are need to confirm this finding.

Diffuse intrinsic pontine glioma (DIPG) is the main cause of brain tumor-related death among children. Until now, there is still a lack of effective therapy with prolonged overall survival for this disease. A typical strategy for preclinical cancer research is to find out the molecular differences between tumor tissue and para-tumor normal tissue, in order to identify potential therapeutic targets. Unfortunately, it is impossible to obtain normal tissue for DIPG because of the vital functions of the pons. Here we report the human fetal hindbrain-derived neural progenitor cells (pontine progenitor cells, PPCs) as normal control cells for DIPG. The PPCs not only harbored similar cell biological and molecular signatures as DIPG glioma stem cells, but also had the potential to be immortalized by the DIPG-specific mutation H3K27M in vitro. These findings provide researchers with a candidate normal control and a potential medicine carrier for preclinical research on DIPG.
Animals ; Brain Stem Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cellular Senescence ; Female ; Glioma ; genetics ; metabolism ; pathology ; Histones ; genetics ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Neoplasm Transplantation ; Neoplastic Stem Cells ; drug effects ; metabolism ; pathology ; Neural Stem Cells ; drug effects ; metabolism ; pathology ; Pons ; embryology ; metabolism ; pathology ; Primary Cell Culture

The existing contents and standards for the standardized residents training are incomplete. Clinical management guidelines and expert consensus should be included in the standardized residents training, since they are very important for clinical decisions. Some guidelines and expert consensus have been included in the standardized residents training in our cardiovascular department,and with the problem-based teaching method, the quality of standardized residents training was improved in the cardiovascular department of ShengJing hospital.

Objective To explore the correlation of total IgE and childhood atopic dermatitis (AD) in maternal serum and newborn cord blood, as well as its clinical significance of allergen testing. Methods Thirty-five cases diagnosed as AD (AD group) were selected, and other 35 children who were not diagnosed as AD (control group) were randomly selected from a birth cohort established in 2009—2011. The total IgE levels were detected by ELISA in maternal serum and newborn cord blood. The serum specific IgE antibody level was detected by quantitative immunoblotting method. Results The serum total IgE level was significantly higher in mother and newborn cord blood in AD group than that in control group (χ2=16.568 and 14.933, P<0.01). Compared to control group, there was a significantly higher positive rate of mother serum allergen includ?ing dust mites, house dust, ragweed pollen, song kind of pollen, poplar, surname and elm pollen, mould, shrimp, marine fish, in AD group (P<0.05). There was a significantly higher positive rate of artemisia pollen and fungi IgE in newborn cord blood in AD group (P<0.05). Conclusion The increased total IgE in maternal serum may play a predictive effect on infants suf?fering from AD. There is no obvious consistency in allergic state between mothers and infants.

Objective To observe the clinical effect , blood glucose time , clearance time of ketone body , correction time of acidosis and influence on average daily dosage of insulin of supplementing Qi and nourishing Yin combined with insulin treatment of patients with diabetic ketoacidosis (DKA).Methods 67 cases of DKA were divided into 2 groups.Control group (n=32) was giv-en micro injection of pump continuous infusion of insulin and aggressive fluid resuscitation for potassium supplement , water and elec-trolyte maintaining and acid -base balance;the observation group ( n=35 ) was given oral Chinese medicine for supplementing Qi and nourishing Yin on the basis of control group .The clinical effect after treatment , blood glucose time , clearance time of ketone body, correction time of acidosis , average daily dosage of insulin and the average number of hypoglycemia per capita were compared and analyzed .Results Blood sugar could be controlled faster in observation group .Clearance time of urine ketone and correction time of acidosis were significantly shortened .The dosage of insulin and number of hypoglycemia per capita were greatly reduced .There were significant differences compared with the control group (p<0.01);in terms of clinical effect, 19 cases of observation group were markedly effective , 14 cases effective , and 2 cases ineffective with the efficiency rate of 94.29%; 13 cases of control group were markedly effective , 10 cases effective , and 9 cases ineffective with the efficiency rate of 71.88%.There were differences between the two groups (p<0.05).Conclusion Supplementing Qi and nourishing Yin can significantly improve the curative effect in the treat -ment of DKA .

Objective To compare the efficacy and safety of intravenous application of amiodarone or esmolol in elderly patients with ventricular arrhythmia following acute myocardial infarction (AMI) within 48 hours,and provide reference for safe use of medication in clinical practice.Methods Elderly hospitalized patients with severe ventricular arrhythmia following AMI in department of Cardiology in Shengjing Hospital of China Medical University from January 2009 to June 2013 were enrolled into the study.The patients were divided into amiodarone group and esmolol group.The patients in amiodarone group received intravenous injection of amiodarone 150 mg in 5％ glucose solution 20 ml within 10 minutes,then an Ⅳ infusion of amiodarone 1 mg/min was given by syringe pump for 6 hours,then an Ⅳ infusion of amiodarone 0.5 mg/min was given for 24-48 hours.After finishing the intravenous application,the patients were given amiodarone orally 0.2 g thrice daily for 7 days.The patients in esmolol group received intravenous infusion of esmolol 0.5 mg/kg within 1 minute,then a venoclysis of esmolol 0.1 mg/(kg · min)was given by syringe pump for 6 hours,then a venoclysis of esmolol 0.05 mg/(kg · min) was given for 24-48 hours.After finishing the intravenous application,the patients were given metoprolol orally 25 mg,twice daily for 7 days.The patient who still had ventricular tachycardia after intravenous application of amiodarone or esmolol for 6 hours in amiodarone group or esmolol group should receive the above-mentioned esmolol or amiodarone treatment,respectively.The efficacy and safety within 48 hours between the 2 groups were compared.Results The effective rate in amiodarone group and esmolol group within 48 hours were 92.1％ (35/38) and 75.0 (27/36),respectively (P =0.046).The incidence of adverse reactions in the 2 groups were 39.5％ (15/38) and 19.4％ (7/36),respectively (P =0.060).Two patients who had accompanied heart failure in the amiodarone group developed acute liver injury 24 hours after intravenous application.Two weeks later,their liver function returned to normal after drug withdrawal and symptomatic treatment.Conclusions The effective rate of intravenous application of amiodarone was higher than that of esmolol in treatment for elderly patients with ventricular arrhythmia following AMI.Amiodarone and esmolol are relatively safe.The liver function should be monitored in the elderly AMI patients when intravenous application of amiodarone were given.Once liver injury occurs,the drug should be discontinued immediately and the symptomatic treatment should be given.

Objective To compare the efficacy and safety of intravenous application of amiodarone or esmolol in elderly patients with ventricular arrhythmia following acute myocardial infarction (AMI) within 48 hours,and provide reference for safe use of medication in clinical practice.Methods Elderly hospitalized patients with severe ventricular arrhythmia following AMI in department of Cardiology in Shengjing Hospital of China Medical University from January 2009 to June 2013 were enrolled into the study.The patients were divided into amiodarone group and esmolol group.The patients in amiodarone group received intravenous injection of amiodarone 150 mg in 5％ glucose solution 20 ml within 10 minutes,then an Ⅳ infusion of amiodarone 1 mg/min was given by syringe pump for 6 hours,then an Ⅳ infusion of amiodarone 0.5 mg/min was given for 24-48 hours.After finishing the intravenous application,the patients were given amiodarone orally 0.2 g thrice daily for 7 days.The patients in esmolol group received intravenous infusion of esmolol 0.5 mg/kg within 1 minute,then a venoclysis of esmolol 0.1 mg/(kg · min)was given by syringe pump for 6 hours,then a venoclysis of esmolol 0.05 mg/(kg · min) was given for 24-48 hours.After finishing the intravenous application,the patients were given metoprolol orally 25 mg,twice daily for 7 days.The patient who still had ventricular tachycardia after intravenous application of amiodarone or esmolol for 6 hours in amiodarone group or esmolol group should receive the above-mentioned esmolol or amiodarone treatment,respectively.The efficacy and safety within 48 hours between the 2 groups were compared.Results The effective rate in amiodarone group and esmolol group within 48 hours were 92.1％ (35/38) and 75.0 (27/36),respectively (P =0.046).The incidence of adverse reactions in the 2 groups were 39.5％ (15/38) and 19.4％ (7/36),respectively (P =0.060).Two patients who had accompanied heart failure in the amiodarone group developed acute liver injury 24 hours after intravenous application.Two weeks later,their liver function returned to normal after drug withdrawal and symptomatic treatment.Conclusions The effective rate of intravenous application of amiodarone was higher than that of esmolol in treatment for elderly patients with ventricular arrhythmia following AMI.Amiodarone and esmolol are relatively safe.The liver function should be monitored in the elderly AMI patients when intravenous application of amiodarone were given.Once liver injury occurs,the drug should be discontinued immediately and the symptomatic treatment should be given.