Objective To understand the awareness of the emergency physicians for thrombolytic thera-py recommended by ST-segment elevation myocardial infarction(STEMI)guidelines and implementation sit-uation,and to analyze the related influencing factors.Methods Relying on Chengdu Emergency Quality Con-trol Center,the questionnaires were distributed to the hospitals of expert group members and the hospitals of the medical union from April to July 2023 to investigate the awareness,implementation status and training status of the emergency doctors in Chengdu area on the STEMI thrombolytic therapy.The causes affecting thrombolysis decision-making were analyzed.Results A total of 137 hospitals participated in the survey.Whether the treatment system and type of chest pain,chest pain center,percutaneous coronary intervention(PCI)qualified hospitals or departments carrying out the training and hospital grade were correlated to whether STEMI patients were thrombolyzed within 12 h before transport(P＜0.05).Whether the PCI quali-fied hospital,chest pain center,chest pain treatment system and type,carrying training,hospital grade,geo-graphical location,transport duration,and thrombolytic indication understanding situation were not related to whether thrombolytic therapy was recommended for the duration from the first medical contact(FMC)to PCI(FMC-PCI)≥120 min(P＞0.05).The multivariate logistic regression analysis showed that whether the de-partment carrying out the training was an influential factor for whether thrombolytic therapy was carried out within 12 h before transport in STEMI onset(P＜0.05).The hospital grade was a influencing factor for whether thrombolysis in non-chest pain treatment system was carried before transport within 12 h of the onset of STEMI.The main reason for STEMI patients being directly transfered treatment without thrombolysis within 12 h of onset was because they knew that thrombolysis was needed but did not know how to do it(50.00％).The awareness rate of thrombolytic indication in non-main urban area was higher than that in main urban area(P＜0.05).The carrying out department training rate of PCI qualified hospitals was higher than that of non-PCI qualified hospitals(P＜0.05).The receiving superior hospital training rate also had difference among different hospital grades and geographical locations.Conclusion Conducting the thrombolysis training and enhancing the learning of treatment processes directly affect the emergency physicians'choice of reperfu-sion strategies for the patients with STEMI.Continuous promotion of the construction of chest pain center or chest pain treatment unit could further increase the early reperfusion rate of STEMI.

Objective:To compare the early and mid-term results of hybrid coronary revascularization (HCR) and minimally invasive multivessel coronary artery bypass grafting (MICS-CABG) in coronary artery disease patients with low left ventricular ejection fraction and non diabetes mellitus, and to explore the indication of HCR and MICS-CABG.Methods:A retrospective cohort analysis of HCR and MICS-CABG cases with preoperative left ventricular ejection fraction less than 0.40, and without diabetes mellitus were conducted in Xijing Hospital from January 2015 to December 2019. 36 cases in HCR group and 17 cases in MICS group were included in this study. For HCR procedure, minimally invasive left internal mammary artery(LIMA) to the left anterior descending artery (LAD) bypass surgery were performed, and followed by percutaneous coronary intervention (PCI) to treat non LAD lesion 1 to 4 weeks later. MICS-CABG procedure was performed through left anterior small thoracotomy minimally invasive direct coronary artery bypass grafting for multiple diseased vessels.Results:The preoperative SYNTAX score in MICS group was significantly higher than that in HCR group ( P<0.05). There was no perioperative death in both groups. Troponin I, postoperative drainage volume, blood transfusion volume and ventilator ventilation time in MICS group were significantly higher than those in HCR group ( P<0.05). After 12 months follow-up, no patient died in both groups. Furthermore, all LIMA grafts were patency. The stenosis rate of drug-eluting stents in HCR group was similar to that of great saphenous vein grafts in MICS group. LVEF and left ventricular end diastolic diameter of both groups were significantly improved 12 months after operation ( P<0.05). Conclusion:HCR and MICS-CABG are minimally invasive and safe treatment for multivessel coronary artery disease patients with low ejection fraction and non diabetese mellitus. The early and mid-term therapeutic effects are satisfactory. If coronary artery lesions other than LAD are suitable for PCI, HCR should be the preferred treatment.

Extracellular vesicles (EV) are membrane structured vesicles containing proteins, lipids and nucleic acids. EVS produced by virus-infected cells are involved in communication between infected and uninfected cells. EV produced by human cytomegalovirus (HCMV) infected cells can promote the transmission and infection of HCMV and escape the host immune response. It can also activate the body′s immune response against HCMV infection. In-depth study of the mechanism and compositional changes of EV produced by HCMV infected cells will contribute to the prevention, diagnosis, and treatment of HCMV. The current research has achieved some results, but they are not deep enough. Future advancements in EV isolation and identification technologies and the reduction of economic costs will contribute to the extensive development and clinical application of the research.

Objective To investigate the application prospect of the most extensive genome engineering pig internationally in preclinical xenotransplantation. Methods Porcine endogenous retrovirus (PERV) knockout combined with 3 major heterologous antigen gene knockouts and 9 humanized genes for inhibition of complement activation, regulation of coagulation disorders, anti-inflammatory and anti-phagocytosis were transferred into a pig (PERV-KO/3-KO/9-TG) as a donor, and the heart, liver and kidney were obtained and transplanted to 3 Rhesus macaque recipients respectively to establish a preclinical research model of pig-to-Rhesus macaque xenotransplantation. The functional status of xenografts after blood flow reconstruction was observed and the survival of recipients was summarized. The hemodynamics of xenografts were monitored. The change of hematological indexes of each recipient was compared. The histopathological manifestation of xenografts was observed. Results After the blood flow was reconstructed, all xenografts showed ruddy color, soft texture and good perfusion. The transplant heart, liver and kidney showed full arterial and venous blood flow and good perfusion at 1 d after operation. The postoperative survival time of heart, liver, and kidney transplant recipients was 7, 26, and 1 d, respectively. The levels of creatine kinase, creatine kinase isoenzyme, and lactate dehydrogenase increased in heart transplant recipient at 1 d after operation, and gradually recovered to near normal levels at 6 d after operation. All indexes increased sharply at 7 d after operation. The level of aspartate aminotransferase increased in liver transplant recipients at 2 d after operation, and the alanine aminotransferase basically returned to normal at 10 d after operation, but the total bilirubin continued to increase. Both aspartate aminotransferase and alanine aminotransferase increased at 12 d after operation, and reached a peak at 15 d after operation. The kidney transplant recipient developed mild proteinuria at 1 d after operation, and died of sudden severe arrhythmia. Histopathology showed that the tissue structure of cardiac and renal xenografts was close to normal, and liver xenografts presented with patchy necrosis, the liver tissue structure was disordered, accompanied by inflammatory damage, interstitial hemorrhage and thrombotic microangiopathy. Conclusions PERV-KO/3-KO/9-TG pig shows advantages in overcoming hyperacute rejection, mitigating humoral rejection and coagulation dysregulation. However, whether it can be used as potential donor for clinical xenotransplantation needs further evaluation.

Circular RNAs (circRNAs) are covalently linked circular single-stranded RNAs, which have the characteristics like abundance,conservation, stabilization and specificity. It is demonstrated that circRNA plays an important roles in the regulation of gene expression and multiple signal pathways of diseases such as cancer, cardiovascular disease, nervous system disease and autoimmune disease. In recent years, it has been reported that circRNAs are expressed abnormally in viral infectious diseases. CircRNA is expected to be a potential biomarker for viral infectious diseases and a potential target for disease therapy. In this review we briefly introduced the research progression of the roles and mechanisms of circular RNAs in viral infectious diseases and provides the references for future study.

Objective To construct an interfering lentivirus vector of long non-coding RNA 4.9 (lncRNA4.9) transcribed by human cytomegalovirus (HCMV).Methods Three interfering sequences targeting lncRNA 4.9 were designed and synthesized.The shuttle plasmid GV248 and the target interfering sequence were combined and constructed.The recombinant plasmid was co-transfected with the skeleton plasmids pHelper1.0 and pHelper2.0 to 293T cells.Viral particles were collected and copies were determined.The interfering lentivirus vector was transfected into THP-1 cells to observe the fluorescence expression,and the interfering efficiency was detected by real-time RT-PCR.Results Three groups of lentivirus interference vectors (LV1,LV2,LV3) were constructed,LV2 and LV3 can interfere with the expression of lncRNA4.9,and the interference efficiency of LV2 group was the highest.Conclusions The interfering lentivirus vector of lncRNA 4.9 was successfully constructed,which laid a foundation for further study on the function of lncRNA 4.9.

The advance of molecular diagnosis has evolved the field of detection of pathogens in pediatric disease towards the high-throughput ultra-sensitive molecular testing. This review provides an overview and comparison of current molecular tools on the market, including qPCR, multiplex PCR, digital PCR, isothermal amplification, and microfluidic chip, highlighting the future trends of integrated high-throughput molecular testing and automation. These evolutions will maximize the utility of limit patient samples from children and contribute significantly on the diagnosis of infectious disease.

Objective To analyze the infectious status and genotype characteristics of group A rotavirus (RV) in children with diarrhea in Hangzhou city in 2017, and to provide information for the disease surveillance, epidemic control as well as vaccine development.Methods Fecal samples from children with acute diarrhea at Children's Hospital of Zhejiang University were collected from Jan to Dec in 2017. All samples were tested for RV antigen by emulsion technique. The antigen-positive samples were further detected by reverse transcription polymerase chain reaction (RT-PCR) and sequencing to determine the G and P genotypes. The RV positive rates in different genders, ages and months were compared by chi-square test. Results A total of 20895 fecal samples were collected from 12389 male patients and 8506 female patients. The gender ratio was 1.46:1. In 5012 (23.99％) RV antigen positive samples, 2964 (23.92％) were from male patients and 2048 (24.08％) were from female patients. There was no gender difference in RV positive rate (χ2=0.049, P>0.05). In the study, RV could be detected in the whole year. January, February and December were peak months, and the RV positive rates were significantly different in different months (χ2=2654.681, P<0.05). The highest RV positive rate was in 18-24 months age group and the lowest in<6 months age group, respectively. Children under 2 years old accounted for 76.56％RV positive cases, and those under 5 years old accounted for 98.72％ RV positive cases. The RV positive rates were also significantly different in all age groups (χ2=1013.832, P<0.05). A total of 116 samples were selected from each month, following the random stratified sampling principle, for PCR amplification, sequencing, and genotyping according to VP7 (G genotype) and VP4 (P genotype). A total of 4 G genotypes were detected and G9 (85.3％) was the predominant one. In the two detected P genotypes, P[8] (96.6％) was predominant. The 4 G/P combination genotypes were G9P[8] (85.3％), G3P[8] (9.5％), G2P[4] (3.5％), and G1P[8] (1.7％). Conclusions RV was a common pathogen in pediatric patients with acute diarrhea in Hangzhou city in 2017. Children under 5 years old, especially 18 months to 2 years old infants was the main infected population. The study showed RV infection had obvious seasonality and winter was the peak period. The G9 genotype was predominant in G genotypes, P[8] genotype was predominant in P genotypes, and G9P[8] genotype was predominant in G/P combination genotypes, respectively.

Objective: To study the relationship between human cytomegalovirus (HCMV) envelope glycoprotein gene H and clinical features of children with congenital cytomegalovirus infection. Methods: A cohort study was conducted. Newborns diagnosed with congenital cytomegalovirus infection, hospitalized in the Department of Neonatology and Neonatal Intensive Care Unit (NICU) of the Children′s Hospital, Zhejiang University School of Medicine, were included from July 2013 to December 2015.HCMV-DNA gH typing in urine, sputum or blood was conducted. Patients then were divided into gH1 group and gH2 group according to gH genotypes. Patients′ data during hospitalization in newborn and 3-5 years of follow-up were collected.The relationships between gH genotype and clinical manifestations, laboratory examinations, hearing loss and neurological prognosis were analyzed by chi-square test, t test and non-parametric test. Results: A total of 21 cases were enrolled as congenital HCMV infection and followed-up for 3-5 years. Among them, 14 (67%) were gH1 type and 7 (33%) were gH2 type. No mixed infection was found. In the two groups, there were no significant differences in the ratio of males (9/14 vs. 3/7,P=0.397), or birth weight ((2 609±686) vs. (3 021±451) g, t=-1.436, P=0.167). Gestational age of gH1 group was younger than that of gH2 group (38 (29-40) vs. 39(38-40) weeks, Z=-2.18, P=0.029). Moderate to severe hearing loss detected by neonatal auditory brainstem response were found in 40 ears (20 cases). It was higher in gH1 group than that in gH2 group (4/22 vs.0/18, χ²=5.145, P=0.023). In the imaging examination of the nervous system, the Alarcon score of gH1 group was lower than that of gH2 group (0.4±0.3 vs. 1.3±1.1, t=-2.459,P=0.024).No significant statistical difference was found in the probability of motor or language development lag in gH2 group and gH1 group (4/7 vs.4/14, P=0.346). Conclusions Compared with gH2 infection, gH1 infection in children has a younger gestational age. The major type of hearing loss in neonatal period is gH1 infection. Children with gH2 congenital infections are more likely to suffer from nervous systems damage.

Gasdermin family (GSDMs), consisting of six proteins (GSDMA, GSDMB, GSDMC, GSDMD, GSDME and DFNB59) in humans and ten proteins (GSDMA1-3, GSDMC1-4, GSDMD, GSDME and DFNB59) in mice, might be involved in multiple physiological and pathological processes, including epithelial cell development, apoptosis, inflammation, carcinogenesis and immune-related diseases. Recent studies confirmed GSDMD, which containing an N-terminal domain with pore-forming activity and a C-terminal domain with structural autoinhibition, as a crucial substrate of inflammatory caspases in pyroptosis, pioneering a new area for structural and functional research on Gasdermin family proteins. This review will summarize the latest progress in the structures, functions and association with diseases of several Gasdermin family proteins.