Objective To investigate the role of Toll-like receptor 4(TLR4)in the regulation of homocys-teine(Hcy)-induced ferroptosis in macrophages.Methods Mouse macrophage cells RAW264.7 were cultured and divided into control group,Hcy intervention group(Hcy group),and Hcy plus ferroptosis inhibitor group(Hcy+Fer-1 group).After transfection with interference fragments,macrophages were treated with Hcy,and then divided into control group,Hcy intervention group(Hcy group),TLR4 interference negative control plus Hcy intervention group(si-NC+Hcy group),and TLR4 interference plus Hcy intervention group(si-TLR4+Hcy group).Macrophages were transfected with overexpression lentivirus and treated with Hcy,then were divided into control group,Hcy intervention group(Hcy group),a TLR4 overexpression negative control plus Hcy intervention group(OE-NC+Hcy group),and a TLR4 overexpression plus Hcy intervention group(OE-TLR4+Hcy group).After 48 hours of intervention,real-time fluorescent quantitative PCR and western blot were used to detect the expression levels of TLR4 in macrophages treated with Hcy;western blot was used to detect the expression levels of ferroptosis-related proteins ACSL4,GPX4,and FTH1 in macrophages,and ferrous ion assay kit to detect the concentration of Fe2+in macrophages;reactive oxygen species(ROS)assay kit and laser confocal microscopy were used to detect the content of intracellular reactive oxygen species.Results Compared with those in the control group,the expression level of the pro-ferroptosis protein ACSL4 was increased in the Hcy group(P<0.05),while the expression levels of anti-ferroptosis proteins GPX4 and FTH1 were decreased(P<0.05);the concentration of Fe2+was increased(P<0.05),and the content of ROS was increased.Meanwhile,the protein and mRNA expres-sion levels of TLR4 were both increased in the Hcy group(P<0.05).After macrophages were transfected with TLR4 interference fragments,compared with those in the si-NC+Hcy group,the expression levels of GPX4 and FTH1 were increased(P<0.05);the expression level of ACSL4 was decreased(P<0.05);the concentration of Fe2+was decreased(P<0.05),and the content of ROS was reduced in the si-TLR4+Hcy group.After macro-phages were transfected with TLR4 overexpression lentivirus,compared with those in the OE-NC+Hcy group,the expression levels of GPX4 and FTH1 were decreased(P<0.05),and the expression level of ACSL4 was increased(P<0.05)in the OE-TLR4+Hcy group.Conclusion Hcy induces the occurrence of ferroptosis in macrophages,and Toll-like receptor 4 has a positive feedback regulatory effect on ferroptosis in macrophages.

Objective To investigate the role of Toll-like receptor 4(TLR4)in the regulation of homocys-teine(Hcy)-induced ferroptosis in macrophages.Methods Mouse macrophage cells RAW264.7 were cultured and divided into control group,Hcy intervention group(Hcy group),and Hcy plus ferroptosis inhibitor group(Hcy+Fer-1 group).After transfection with interference fragments,macrophages were treated with Hcy,and then divided into control group,Hcy intervention group(Hcy group),TLR4 interference negative control plus Hcy intervention group(si-NC+Hcy group),and TLR4 interference plus Hcy intervention group(si-TLR4+Hcy group).Macrophages were transfected with overexpression lentivirus and treated with Hcy,then were divided into control group,Hcy intervention group(Hcy group),a TLR4 overexpression negative control plus Hcy intervention group(OE-NC+Hcy group),and a TLR4 overexpression plus Hcy intervention group(OE-TLR4+Hcy group).After 48 hours of intervention,real-time fluorescent quantitative PCR and western blot were used to detect the expression levels of TLR4 in macrophages treated with Hcy;western blot was used to detect the expression levels of ferroptosis-related proteins ACSL4,GPX4,and FTH1 in macrophages,and ferrous ion assay kit to detect the concentration of Fe2+in macrophages;reactive oxygen species(ROS)assay kit and laser confocal microscopy were used to detect the content of intracellular reactive oxygen species.Results Compared with those in the control group,the expression level of the pro-ferroptosis protein ACSL4 was increased in the Hcy group(P<0.05),while the expression levels of anti-ferroptosis proteins GPX4 and FTH1 were decreased(P<0.05);the concentration of Fe2+was increased(P<0.05),and the content of ROS was increased.Meanwhile,the protein and mRNA expres-sion levels of TLR4 were both increased in the Hcy group(P<0.05).After macrophages were transfected with TLR4 interference fragments,compared with those in the si-NC+Hcy group,the expression levels of GPX4 and FTH1 were increased(P<0.05);the expression level of ACSL4 was decreased(P<0.05);the concentration of Fe2+was decreased(P<0.05),and the content of ROS was reduced in the si-TLR4+Hcy group.After macro-phages were transfected with TLR4 overexpression lentivirus,compared with those in the OE-NC+Hcy group,the expression levels of GPX4 and FTH1 were decreased(P<0.05),and the expression level of ACSL4 was increased(P<0.05)in the OE-TLR4+Hcy group.Conclusion Hcy induces the occurrence of ferroptosis in macrophages,and Toll-like receptor 4 has a positive feedback regulatory effect on ferroptosis in macrophages.

Objective:To summarize the clinical treatment of complete left bundle branch block (CLBBB) after the transcatheter closure of ventricular septal defect (VSD).Methods:A case series study was conducted on the treatments and outcomes of 25 children with CLBBB after transcatheter VSD closure in Guangdong Provincial People′s Hospital from January 2010 to December 2023.Paired sample t test was used to evaluate the effect of occlude removal. Results:Among the 25 patients, 12 were males (48%), and 13 were females (52%).The age at surgery was 3.18 (2.51-3.86) years, the height before surgery was 95.0 (90.0-97.5) cm, and the weight before surgery was 13 (12-15) kg.Fourteen children were early-onset cases (≤ 1 month), while the other 11 were late-onset cases (> 1 month).The mean follow-up time was (6.63±3.93) years.Of the 14 early-onset cases, 6 children underwent occluder removal within 1 month and restored normal heart rhythm or incomplete right bundle branch block; 4 children underwent occluder removal after 1 month, of whom 2 recovered, 1 remained CLBBB, and 1 had complete atrioventricular block (CAVB); the other 4 children received drug treatment, of whom 2 had normal heart rhythm, 1 had left anterior fascicular block, and 1 died of cardiac shock and heart failure.All the 11 late-onset cases were first treated by drugs, of whom 3 recovered, and the other 8 remained CLBBB.One of the 8 cases received occluder removal at 8 months after surgery and recovered, 1 had CAVB, and the other 6 remained CLBBB.Conclusions:For patients with CLBBB after transcatheter closure of VSD, drug therapy is not always effective, and CLBBB is easy to recur.Therefore, occluder removal is recommended to be done immediately after CLBBB is discovered.Patients with persistent CLBBB should be followed up regularly, and pacemaker implantation may be performed if necessary.

Objective:To evaluate the implementation of national continuing medical education (CME) base programme about infectious disease control and prevention during 2013-2020, so as to improve the quality management of CME.Methods:According to data from national CME system, Excel and SPSS 27.0 were used to analyze project hosting days, places, teachers, students, project directors and training effect. The counting data were expressed by frequency and percentage [cases (%)], chi-square test was used to make comparison between groups, Mantel-Haenszel chi-square test was used for trend test, and the significance test level of the difference was α = 0.05. Results:A total of 116 projects were conducted from 2013 to 2020, with execution rate of 87.9%(116/132). Most hosting days were 2 to 3 days [57.8% (67/116)]. The majority [65.2% (5 785/8 871)] of trainees had junior and intermediate technical titles. As for trainers, trainers with senior technical titles accounted for 87.6% (758/865), and those with intermediate titles accounted for 12.4% (107/865). Mantel-Haenszel chi-square test showed that there was a linear relationship between the proportion of technical titles and the year ( χ2趋势 = 4.97, P趋势 = 0.026). Project directors almost had senior professional title, and nearly one third of them had the experience of undertaking three or more base projects within 8 years. The top three training modules were parasitic diseases prevention and control, AIDS prevention and control, and viral diseases prevention and control. Trainees were highly satisfied with the training contents. Conclusion:The implementation of the infectious disease prevention and control base programme went well in general from 2013 to 2020. In the future, it’s needed to be demand-oriented, rationally design training programs, enhance the evaluation of training effects, strengthen the construction of public health core capacity, and adopt a strategy of brand development in the process of the infectious disease prevention and control base programme.

Objective To establish a PCR-based capillary electrophoresis(PCR/CE)to detect Survival Motor Neuron 1(SMN1)and Survival Motor Neuron 2(SMN2)genes and to evaluate its performance.Methods PCR/CE and Multiplex Ligation-dependent Probe Amplification(MLPA)for SMA gene diagnosis were used to blindly test the samples in sync.The performance of PCR/CE was assessed using MLPA results as the standard.Results A total of 336 samples were included in this study,consisting of 50 homozygous deletion types(14.9%),65 heterozygous deletion types(19.3%),and 221 non-deletion types(65.8%).The results of PCR/CE for detect-ing SMN1 and SMN2 copy numbers(0,1,2,3,≥4)were in complete agreement with the results of the MLPA.Conclusions PCR/CE for gene testing related to SMA could accurately detect copy numbers of exon 7 and exon 8 of the SMN1 and SMN2 genes(0,1,2,3,≥4).

Objective:To investigate the value of single sperm sequencing combined with PCR-reverse dot blot (PCR-RDB) technology in preimplantation genetic testing (PGT) of Southeast Asian deletion type α thalassemia.Methods:A couple of Southeast Asian deletion type α thalassemia carrier was selected in this case, who asked for assisted reproduction in the Department of Obstetrics and Gynecology of the Third Affiliated Hospital of Guangzhou Medical University on April 24, 2020. The male carrier was subjected to single sperm isolation by the swim-up method combined with micropipette due to his incomplete pedigree. Five single sperm samples were isolated and their whole-genome were amplified. The genotype of thalassemia of single sperm samples were determined by PCR-RDB. SNPs were selected as genetic markers in the range of 1 Mb upstream and downstream of -- SEA region to construct chromosomal haplotype. Next, we performed whole-genome amplification on six blastocyst biopsy samples, and then next-generation sequencing was carried out to detect the carrier status of the embryos. Finally, the non-pathogenic blastocysts were selected for transplantation, and the preimplantation genetic testing for monogenic (PGT-M) results were confirmed by the prenatal genetic diagnosis during pregnancy. Results:The pathogenic variant in the female was inherited from her mother, and four of the five single sperm samples were wild-type. Ten male informative SNP markers were identified by single sperm sequencing, and six female informative SNP markers were identified by linkage analysis of female family. PGT-M results indicated that four blastocysts were αα/-- SEA and two were -- SEA/-- SEA. The result of prenatal diagnosis revealed that the fetus was a carrier of -- SEA , which was consistent with PGT-M result, and a healthy girl was delivered at 40 weeks of gestation. Conclusion:For male carriers of Southeast Asian deletion type α thalassemia with incomplete pedigree, single sperm sequencing combined with PCR-RDB technology can be used to select SNP sites, and PGT-M can be performed by linkage analysis.

Objective:To investigate the value of single sperm sequencing combined with PCR-reverse dot blot (PCR-RDB) technology in preimplantation genetic testing (PGT) of Southeast Asian deletion type α thalassemia.Methods:A couple of Southeast Asian deletion type α thalassemia carrier was selected in this case, who asked for assisted reproduction in the Department of Obstetrics and Gynecology of the Third Affiliated Hospital of Guangzhou Medical University on April 24, 2020. The male carrier was subjected to single sperm isolation by the swim-up method combined with micropipette due to his incomplete pedigree. Five single sperm samples were isolated and their whole-genome were amplified. The genotype of thalassemia of single sperm samples were determined by PCR-RDB. SNPs were selected as genetic markers in the range of 1 Mb upstream and downstream of -- SEA region to construct chromosomal haplotype. Next, we performed whole-genome amplification on six blastocyst biopsy samples, and then next-generation sequencing was carried out to detect the carrier status of the embryos. Finally, the non-pathogenic blastocysts were selected for transplantation, and the preimplantation genetic testing for monogenic (PGT-M) results were confirmed by the prenatal genetic diagnosis during pregnancy. Results:The pathogenic variant in the female was inherited from her mother, and four of the five single sperm samples were wild-type. Ten male informative SNP markers were identified by single sperm sequencing, and six female informative SNP markers were identified by linkage analysis of female family. PGT-M results indicated that four blastocysts were αα/-- SEA and two were -- SEA/-- SEA. The result of prenatal diagnosis revealed that the fetus was a carrier of -- SEA , which was consistent with PGT-M result, and a healthy girl was delivered at 40 weeks of gestation. Conclusion:For male carriers of Southeast Asian deletion type α thalassemia with incomplete pedigree, single sperm sequencing combined with PCR-RDB technology can be used to select SNP sites, and PGT-M can be performed by linkage analysis.

Objective:To investigate the immunogenicity and safety of a boost dose of measles, mumps, and rubella combined vaccine (MMR) for children 4 to 6 years old.Methods:Children, aged 4 to 6 years old, had vaccinated with 1 dose of measles and rubella combined vaccine(MR) at the age of 8 months and 1 dose of MMR vaccine at 18-months, were recruited in Shanxi, Inner Mongolia, and Beijing, respectively. All children were assigned into 4, 5 and 6-year-old group. The children who met inclusion and exclusion criteria were vaccinated with 1 dose MMR vaccine, and were collected blood samples before vaccination and 35 to 42 d after the vaccination. During the study period, adverse events were collected at 30 min, 1 d, 2 d, 3 d, 4-12 d, and 13 to 42 days after vaccination. Serum was tested for IgG antibodies against measles, mumps and rubella. Geometric mean concentrations (GMC) of measles, mumps, and rubella antibodies were compared among groups by analysis of variance or non-parametric test. Seropositive rates and adverse event rates were compared among groups by Chi-square test or Fisher exact test.Results:A total of 500 children were included in immunogenicity analysis and 535 children were included in safety analysis. The overall adverse event rate was 20.37%, the most of severity for adverse events was mild. The rates of local and systemic adverse events were 0.37% and 20.00%, respectively. Symptoms of local adverse events were redness. The main systemic adverse events were fever, followed by cough, rash and runny nose. Received a dose of MMR vaccine for booster immunization, the seropositive rates of measles antibody, mumps antibody and rubella antibody were above 99% for all 3 age groups, and there was no significant difference between groups. There were significant differences in mumps antibody GMC among groups ( P=0.042), but no significant differences in measles and rubella antibodies GMC. Conclusion:The immunogenicity and safety of a boosted MMR vaccintion in children aged 4, 5 and 6 years were all similar good.

Objective To evaluate the expression of human vascular endothelial cell growth factors 165 (hVEGF165) gene transfected into fibroblasts by recombinant adenovirus and study the repairing effect of this cells on radiated skin ulcer in rats.Methods The recombinant adenovirus with hVEGF165 was established and transfected to rat primary fibroblasts, and its expression of hVEGF165 in fibroblasts was identified with real-time PCR, immunocytochemistry and Western blot.Twenty four clean grade SD rats of were irradiated locally with 50 Gy γ rays to generate an animal model of radiation skin injury.The hVEGF165-transfected cells were injected to the irradiated site under rat skin 7 d post-irradiation.The therapeutic effects on the irradiated skin wound were evaluated through general observation as well as histological staining of HE.The expression of hVEGF in the irradiated skin tissue with fibroblasts injection was analyzed by Real-time PCR.Results The hVEGF165 gene was overexpressed in the transfected cells and approached to 88 373-fold bigger compared to controls transfected with blank vectors, and an extensive expression of VEGF in the cytoplasm of transfected cells was observed by immunohistochemistry.VEGF protein with the relative molecular mass of 23 000 was also detected in cell lysate by Western blot.The local skin ulcers in rats occurred about two weeks after irradiation.In the hVEGF165-transfected group, the average area of radiation-injured skin was 40.2 mm2, about 57％ less than that of the control group transfected with blank vectors so that the healing time was shorten by 6 days.The relative concentration of hVEGF mRNA in the skin tissue of rats injected with hVEGF165-transfected cells were 5.15-fold and 4.15-fold bigger compared to that of controls (t =3.385,3.220, P ＜ 0.05) at 3 and 7 d after administration.Conclusions The primary fibroblasts transfected with hVEGF165 gene could efficiently release VEGF to the irradiated skin tissue and promote the recovery of irradiation skin ulcers by shortening the healing time and thus enhanced the therapeutic effect on skin wounds.

Maximum a Posteriori (MAP) method has been widely applied to the ill-posed problem of image reconstruction. The choice of prior is the crucial point on MAP methods. However, the most conventional priors will lead to a blurring of the whole image or cause ladder-like artifacts. We therefore proposed a Tsallis entropy-based prior for positron emission tomography (PET) iterative reconstruction in MAP framework. The method uses a Tsallis entropy-based prior to eliminate the uncertainty between prior information and the estimated images. We tested this method in the phantom image, compared it with the traditional prior methods. the results showed that the proposed algorithm could suppress noise and obtain better reconstructed image quality.
Algorithms ; Artifacts ; Entropy ; Image Processing, Computer-Assisted ; methods ; Phantoms, Imaging ; Positron-Emission Tomography ; methods