This study investigated the specific immune response of BALB/c mice that was induced by a circular RNA (circRNA) vaccine expressing the herpes simplex virus type II (HSV-2) glycoprotein D (gD). The aim was to evaluate the immunological potential of this vaccine and lay a foundation for developing an mRNA vaccine against HSV-2. PCR and homologous recombination were employed to integrate the gD gene obtained from the pT7AMP-gD ectodomain plasmid into pUC57 to generate the recombinant plasmid pUC57-circ-gD, which was then sequenced and characterized. In vitro transcription and cyclization were performed on the template DNA to generate pUC57-circ-gD mRNA. To validate the formation of circular RNA, we cleaved the pUC57-circ-gD mRNA with RNase R and employed RT-PCR to validate the cyclization. The pUC57-circ-gD mRNA was then transfected into 293T cells. After 72 h, the cell supernatant was collected, and Western blotting was employed to measure the protein level of gD. Subsequently, the mRNA was encapsulated in lipid nanoparticles (LNPs) by microfluidic encapsulation. BALB/c mice were administrated with the encapsulated mRNA, and blood was collected from the fundus venous plexus after 21 and 35 days, and from the enucleated eyeballs after 49 days. Enzyme-linked immunosorbent assay was employed to measure the titers of antibodies, including virus-neutralizing antibodies. After 49 days, spleens were harvested and assessed for secretion of interferon-gamma (IFN-γ) by solid-phase enzyme-linked immunospot. The results showed successful construction and sequencing of the recombinant plasmid. RNase R digestion confirmed the presence of circular RNAs. Western blotting of the 293T cells transfected with the mRNA showed clear specific bands. The quality of the vaccine was tested by size exclusion chromatography-high performance liquid chromatography, which showed that the purity of the vaccine was about 90%. The mRNA-LNP showcased the particle size of 82.76 nm and an encapsulation rate of approximately 98%. Following three-dose vaccination, all immunized mice exhibited steady weight gain with 100% survival rate throughout the 28-day observation period, indicating no significant acute toxicity associated with the vaccine formulation. The immunized mice showed dose-dependent increases in serum IgG antibody titer and IFN-γ secretion by splenocytes and they were resistant to virus attacks. These findings indicate good immunogenicity and persistence of the pUC57-circ-gD mRNA vaccine, providing a reference for further studies on circRNA vaccines.
Animals ; Mice, Inbred BALB C ; RNA, Circular ; Mice ; Humans ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Antibodies, Viral/blood* ; HEK293 Cells ; Female ; Nanoparticles ; Plasmids

ObjectiveTo understand the epidemic characteristics of influenza in Beijing from 2023 to 2024, and to provide a scientific basis for the prevention and control of influenza. MethodsData on influenza-like illness (ILI) from secondary level and above hospitals, etiology surveillance data, and influenza clusters outbreaks data from 2023‒2024 were used to analyze the epidemic trend and pathogenic characteristics of influenza. Furthermore, an influenza comprehensive index was used to categorize the epidemic intensity at the severity level. ResultsA total of 2 065 857 ILI cases were reported in 2023‒2024 epidemic season, and the percentage of ILI was 3.67%. The age group of 5‒14 years accounted for the highest proportion of ILI (30.48%). A total of 41 766 throat swabs from ILI were detected, with a positive rate of 17.28%.A (H3N2) (51.86%) and B Victoria (41.93%) were the most prevalent subtypes of influenza virus. Clustered influenza outbreaks occurred mainly in primary schools (57.78%) and middle schools (35.55%), mainly caused by the influenza A (H3N2) subtype (85.93%). According to the influenza comprehensive index (I), the period of influenza activity and above (I>0.5) lasted for a total of 37 weeks, accounting for 71.15% of the entire influenza season. ConclusionCompared with previous years, the epidemic level of influenza in Beijing was increased in 2023‒2024, and the peak time became earlier. The comprehensive index method can objectively evaluate the level of influenza epidemic and provide suggestions for the future prevention and control of influenza in Beijing.

Objective:To analyze the effect of influenza vaccination on influenza cluster epidemic in primary and secondary schools in Beijing during the 2023-2024 surveillance season and provide evidence for the improvement of influenza vaccination strategies.Methods:The incidence data of influenza cluster epidemic and influenza vaccination coverage in the schools in Beijing during 2023-2024 were collected. Descriptive epidemiological methods were used to analyze cluster epidemic characteristics, and χ2 test was used to compare incidence differences between groups, and OR value and vaccine effectiveness [VE=(1- OR)×100%] were calculated. A negative binomial regression model was used to evaluate the association between school vaccination rates and cluster epidemic risk. Joinpoint regression was used to analyze trends in relative risk ( RR) with increasing vaccination coverage and to determine the optimal vaccination threshold. Results:A total of 126 influenza cluster epidemic were reported in 115 primary and secondary schools in Beijing during 2023-2024 with the median size of 15 case, the average attack rate was 36.26% (2 033/5 607). The epidemics mainly occurred in urban area (70, 55.56%). Primary schools were the main setting (78, 61.90%), and influenza A(H3N2) was the predominant subtype (108, 85.71%). The overall influenza vaccination coverage in the primary and secondary students was 54.26%, while the average vaccination in classes affected by the epidemics was 58.57%. The overall protection rate was 47.62%, the protection rate was higher in primary schools (49.65%) than in secondary schools (46.60%). The protection rates against influenza A(H1N1)pdm09 (80.93%) and influenza B (Victoria lineage) (81.65%) were significantly higher than that against influenza A(H3N2) (44.19%). When school vaccination coverage reached ≥76.00%, the epidemic risk decreased by 52.82%.Conclusions:Even the match between influenza vaccine strains and circulating strains is suboptimal, increasing influenza vaccination coverage in schools can effectively reduce the risk for influenza cluster epidemic. In the future, measures such as policy guidance and public health education should be taken to further improve vaccination coverage, thereby establishing herd immunity and reducing the transmission risk of influenza in schools.

Ventilator-induced lung injury(VILI)is a form of lung injury caused by inappropriate ventilator parameter settings or pre-existing lung pathologies during mechanical ventilation.The pathogenesis of VILI is complex,primarily involving barotrauma,volutrauma,atelectrauma,and biotrauma.To prevent VILI,clinical practice advocates protective lung ventilation strategies and personalized positive end-expiratory pressure(PEEP)settings.Early identification and precise assessment are critical for improving patient outcomes.The rapid advancement of imaging technologies has provided multidimensional perspectives for dynamic monitoring of VILI.This review summarizes the imaging principles,clinical applications in the context of VILI management,and limitations of various imaging modalities,including lung ultrasound(LUS),computed tomography(CT),electrical impedance tomography(EIT),and magnetic resonance imaging(MRI).By offering comprehensive diagnostic information,this review aims to provide a theoretical foundation for the precise assessment of VILI and the optimization of ventilatory support strategies.

Objective:To analyze the effect of influenza vaccination on influenza cluster epidemic in primary and secondary schools in Beijing during the 2023-2024 surveillance season and provide evidence for the improvement of influenza vaccination strategies.Methods:The incidence data of influenza cluster epidemic and influenza vaccination coverage in the schools in Beijing during 2023-2024 were collected. Descriptive epidemiological methods were used to analyze cluster epidemic characteristics, and χ2 test was used to compare incidence differences between groups, and OR value and vaccine effectiveness [VE=(1- OR)×100%] were calculated. A negative binomial regression model was used to evaluate the association between school vaccination rates and cluster epidemic risk. Joinpoint regression was used to analyze trends in relative risk ( RR) with increasing vaccination coverage and to determine the optimal vaccination threshold. Results:A total of 126 influenza cluster epidemic were reported in 115 primary and secondary schools in Beijing during 2023-2024 with the median size of 15 case, the average attack rate was 36.26% (2 033/5 607). The epidemics mainly occurred in urban area (70, 55.56%). Primary schools were the main setting (78, 61.90%), and influenza A(H3N2) was the predominant subtype (108, 85.71%). The overall influenza vaccination coverage in the primary and secondary students was 54.26%, while the average vaccination in classes affected by the epidemics was 58.57%. The overall protection rate was 47.62%, the protection rate was higher in primary schools (49.65%) than in secondary schools (46.60%). The protection rates against influenza A(H1N1)pdm09 (80.93%) and influenza B (Victoria lineage) (81.65%) were significantly higher than that against influenza A(H3N2) (44.19%). When school vaccination coverage reached ≥76.00%, the epidemic risk decreased by 52.82%.Conclusions:Even the match between influenza vaccine strains and circulating strains is suboptimal, increasing influenza vaccination coverage in schools can effectively reduce the risk for influenza cluster epidemic. In the future, measures such as policy guidance and public health education should be taken to further improve vaccination coverage, thereby establishing herd immunity and reducing the transmission risk of influenza in schools.

Objective:To investigate the current status and challenges of pediatric life support training in China and provide references for improving training quality.Methods:A cross-sectional study was conducted to collect data from pediatric life support training centers across the country，covering basic institutional information，training capacity and training faculty，training program funding，as well as existing challenges and issues.The domestic registry of training centers in 2023 was obtained through the American Heart Association's online platform.After contacting and verifying each center，an online questionnaire was distributed，and the aggregated data were statistically analyzed.Results:A total of 42 institutions participated in the survey，including 19 children's hospitals，14 general hospitals，6 maternal and child health hospitals，2 women and children’s hospitals，and 1 training institution.The distribution of training centers showed a concentration in coastal areas，with the top three provinces/municipalities being Guangdong（7/42，16.7%），Zhejiang（6/42，14.3%），and Shanghai（4/42，9.5%）.As of December 31 2023，the 42 institutions had an annual basic life support（BLS）training volume of 8 587 individuals，the median was 120 (100,200)，and an annual pediatric advanced life support（PALS）training volume of 2 448 individuals，the median was 30 (20,50).Among the 42 institutions，there were 598 BLS instructors and 306 PALS instructors.Among the surveyed institutions，24（24/42，57.1%）reported BLS instructor teams comprising fewer than 10 members，and 33（33/42，78.6%）reported PALS instructor teams comprising fewer than 10.Only 7 centers(7/42,16.7%）reported having dedicated funding support.The top three challenges were：training sessions occupying instructors’personal time（27/42，64.3%），low instructor compensation（16/42，38.1%），and issues with the data submission system（16/42，38.1%）.Conclusion:Pediatric life support training centers in China are primarily children’s hospitals，with a geographical concentration in coastal areas，which is also reflected in the distribution of training scale and instructor resources.Most centers have relatively small training scales and limited instructor capacity，with many instructors conducting training during their personal time.These issues may hinder the implementation and effectiveness of training programs.

Objective:To investigate the current status and challenges of pediatric life support training in China and provide references for improving training quality.Methods:A cross-sectional study was conducted to collect data from pediatric life support training centers across the country，covering basic institutional information，training capacity and training faculty，training program funding，as well as existing challenges and issues.The domestic registry of training centers in 2023 was obtained through the American Heart Association's online platform.After contacting and verifying each center，an online questionnaire was distributed，and the aggregated data were statistically analyzed.Results:A total of 42 institutions participated in the survey，including 19 children's hospitals，14 general hospitals，6 maternal and child health hospitals，2 women and children’s hospitals，and 1 training institution.The distribution of training centers showed a concentration in coastal areas，with the top three provinces/municipalities being Guangdong（7/42，16.7%），Zhejiang（6/42，14.3%），and Shanghai（4/42，9.5%）.As of December 31 2023，the 42 institutions had an annual basic life support（BLS）training volume of 8 587 individuals，the median was 120 (100,200)，and an annual pediatric advanced life support（PALS）training volume of 2 448 individuals，the median was 30 (20,50).Among the 42 institutions，there were 598 BLS instructors and 306 PALS instructors.Among the surveyed institutions，24（24/42，57.1%）reported BLS instructor teams comprising fewer than 10 members，and 33（33/42，78.6%）reported PALS instructor teams comprising fewer than 10.Only 7 centers(7/42,16.7%）reported having dedicated funding support.The top three challenges were：training sessions occupying instructors’personal time（27/42，64.3%），low instructor compensation（16/42，38.1%），and issues with the data submission system（16/42，38.1%）.Conclusion:Pediatric life support training centers in China are primarily children’s hospitals，with a geographical concentration in coastal areas，which is also reflected in the distribution of training scale and instructor resources.Most centers have relatively small training scales and limited instructor capacity，with many instructors conducting training during their personal time.These issues may hinder the implementation and effectiveness of training programs.

Ventilator-induced lung injury(VILI)is a form of lung injury caused by inappropriate ventilator parameter settings or pre-existing lung pathologies during mechanical ventilation.The pathogenesis of VILI is complex,primarily involving barotrauma,volutrauma,atelectrauma,and biotrauma.To prevent VILI,clinical practice advocates protective lung ventilation strategies and personalized positive end-expiratory pressure(PEEP)settings.Early identification and precise assessment are critical for improving patient outcomes.The rapid advancement of imaging technologies has provided multidimensional perspectives for dynamic monitoring of VILI.This review summarizes the imaging principles,clinical applications in the context of VILI management,and limitations of various imaging modalities,including lung ultrasound(LUS),computed tomography(CT),electrical impedance tomography(EIT),and magnetic resonance imaging(MRI).By offering comprehensive diagnostic information,this review aims to provide a theoretical foundation for the precise assessment of VILI and the optimization of ventilatory support strategies.

Objective:To investigate the relationship between thyroid hormone levels during pregnancy and neonatal thyroid function.Methods:Forty pregnant women with abnormal thyroid stimulating hormone (TSH) level during pregnancy and their newborns who received treatment in Yongkang First People's Hospital, China between July 2019 and August 2020 were included in the observation group. An additional 38 healthy pregnant women and their newborns who concurrently received health examination were included in the control group. The clinical data in the two groups were retrospectively analyzed. The levels of thyroid hormones [(triiodothyronine (T 3), tetraiodothyronine (T 4), TSH, free T 3 (FT 3), free T 4 (FT 4)] in pregnant women were compared between the two groups. TSH level in newborns was compared between the two groups. Thyroid dysfunction in newborns was assessed in each group. The correlation between thyroid hormone levels during pregnancy and neonatal thyroid function was analyzed. Results:TSH level during pregnancy in the observation group was significantly higher than that in the control group [(2.89 ± 0.44) mU/L vs. (2.13 ± 0.22) mU/L, t = 9.570, P < 0.001]. T 3, T 4, FT 3 and FT 4 in the observation group were (0.45 ± 0.07) μg/L, (90.87 ± 8.93) μg/L, (1.08 ± 0.19) ng/L and (10.45 ± 1.73) ng/L, respectively, which were significantly lower than those in the control group [(2.13 ± 0.22) μg/L, (1.31 ± 0.21) μg/L, (2.16 ± 0.34) ng/L, (15.31 ± 21) ng/L, t = 24.514, 9.254, 17.432, 10.845, all P < 0.001]. TSH level in newborns in the observation group was significantly higher than that in the control group ( t = 37.041, P < 0.05). The incidence of thyroid dysfunction in the observation group was significantly higher than that in the control group ( χ2 = 4.780, P < 0.05). TSH level in pregnant women was positively correlated with that in newborns ( r = 0.819, P < 0.05). T 3, T 4, FT 3 and FT 4 levels in pregnant women were negatively correlated with TSH level in newborns ( r = -0.773, -0.802, -0.794, -0.824, all P < 0.05). Conclusion:Compared with healthy pregnant women, pregnant women with abnormal thyroid hormone levels have higher TSH levels and lower T 3, T 4, FT 3 and FT 4 levels. The newborns of pregnant women with abnormal thyroid hormone levels have higher TSH levels and a greater risk of thyroid dysfunction than the newborns of healthy pregnant women. The level of thyroid hormone during pregnancy is related to the thyroid function of newborns. This study is scientific and innovative.

Objective:Assess the relationship between elevated antiphospholipid antibodies and thrombosis in hospitalized patients.Methods:Case control study. A total of 385 patients (149 males and 236 females, aged from 1 to 105 years, with a median age of 37 years) who were hospitalized in Peking University First Hospital from January 2015 to December 2019 and tested positive for any one of the anti-phospholipid antibodies were included in the study. All subjects were divided into thrombotic group and non-thrombotic group according to whether thrombus was detected by imaging examination during hospitalization. In thrombosis group, there were 66 males and 36 females, aged from 3 to 105 years, with a median age of 58 years. In non-thrombosis group, there were 83 males and 200 females, aged from 1 to 94 years, with a median age of 31 years. Clinical data and laboratory data of patients were recorded. ACL-IgM/IgG and anti-β2GPI-IgM/IgG were detected by ELISA and LA was detected by dRVVT and SCT on automatic coagulation analyzer. The rates of age, gender, smoking, obesity, hypertension, hyperlipidemia, diabetes and the median level of antiphospholipid antibodies were compared between two groups. Logistic multivariate regression analysis was used to determine the risk factors for thrombotic events. The mid-to-high titer value of aCL was established by the χ 2-trend test and verified by logistic regression. Results:The median age (58 years) and the rates of male (64.7%), smoking (16.7%), hypertension (63.7%) and diabetes (28.4%) in thrombus group were significantly higher than those in non-thrombus group ( Z=7.685, χ2=38.077, 16.312, 37.769, 24.749 respectively; P<0.01). The positive rate of anti-β2GPI-IgG and dRVVT in thrombosis group (11.8% and 78.4%) was significantly higher than that in non-thrombosis group (5.3% and 60.1%), as well as the median level of dRVVT (1.29 RU/ml vs 1.23 RU/ml) (χ2=3.864 and 10.309, Z=3.539; P<0.05). The median level of aCL-IgM was higher in non-thrombosis group (2.3 MPL vs 2.0 MPL). The positive rate of aCL-IgG was slightly higher in thrombosis group (18.6% vs 10.6%). Logistic regression analysis showed that men, hypertension, diabetes, advanced age, elevated dRVVT, and elevated anti-β2GPI-IgG are risk factors for thrombosis. Taking 36 GPL as the medium-to-high titer value of aCL-IgG, the risk of thrombosis increased by 2.45 times. Conclusions:In the anti-phospholipid antibody profile, LA detected by dRVVT method, anti-β2GPI-IgG and aCL-IgG may be valuable laboratory indicators for inpatient thrombotic events. The mid-to-high titer value of aCL-IgG is set at 36 GPL to distinguish the risk of thrombosis.