ObjectiveTo develop a classification model based on complete blood count （CBC） parameters combined with clinical factors to predict severe respiratory infections caused by Human adenovirus （HAdV） in pediatric patients. MethodsFrom September 2023 to September 2024， the CBC parameters and related clinical data from pediatric patients diagnosed with HAdV infection were collected. Principal component analysis and random forest models were used to identify potential predictors of severe cases. ResultsA total of 668 pediatric patients were included， with 564 cases assigned to the training cohort and 104 cases to the validation cohort. Severe cases were defined as pneumonia and/or fever lasting ≥5 days （pneumonia or prolonged fever， PorPF）. Principal component analysis and feature importance analysis （Mean Decrease Gini value） identified the monocytosis ratio （PMono）， red blood cell count （RBC）， and platelet count （PLT） as the most critical CBC parameters. Logistic regression analysis revealed that oxygen therapy （OR = 4.367， 95% CI： 1.568–12.161） and increased work of breathing （OR = 3.904， 95% CI： 2.146–7.101） were relative risk factors for PorPF. Meanwhile， higher PMono （OR = 0.696， 95% CI： 0.640–0.757）， RBC （OR = 0.201， 95% CI： 0.124–0.325）， and PLT （OR = 0.990， 95% CI： 0.987–0.994） were protective factors. When PMono was used as a predictive marker for PorPF， the area under the receiver operating characteristic curve （AUC） was 0.648 and 0.705， respectively. A random forest model incorporating four risk factors ［PMono， RBC， PLT， and hematocrit （HCT）］ was constructed to classify PorPF and general cases， achieving AUCs of 0.688 and 0.768， respectively. ConclusionsPMono， RBC， and PLT may serve as characteristic CBC indicators for predicting pneumonia or prolonged fever in children with HAdV infection. A risk factor model built using PMono， RBC， PLT， and HCT offers a relatively simple and accurate approach to predicting severe cases in pediatric HAdV infections.

Background::T-cell-mediated immunity is crucial for the effective clearance of viral infection, but the T-cell-mediated immune responses that are induced by booster doses of inactivated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines in people living with human immunodeficiency virus (PLWH) remain unclear.Methods::Forty-five PLWH who had received antiretroviral therapy (ART) for more than two years and 29 healthy controls (HCs) at Beijing Youan Hospital were enrolled to assess the dynamic changes in T-cell responses between the day before the third vaccine dose (week 0) and 4 or 12 weeks (week 4 or week 12) after receiving the third dose of inactivated SARS-CoV-2 vaccine. Flow cytometry, enzyme-linked immunospot (ELISpot), and multiplex cytokines profiling were used to assess T-cell responses at the three timepoints in this study.Results::The results of the ELISpot and activation-induced marker (AIM) assays showed that SARS-CoV-2-specific T-cell responses were increased in both PLWH and HCs after the third dose of the inactivated SARS-CoV-2 vaccine, and a similar magnitude of immune response was induced against the Omicron (B.1.1.529) variant compared to the wild-type strain. In detail, spike-specific T-cell responses (measured by the ELISpot assay for interferon γ [IFN-γ] release) in both PLWH and HCs significantly increased in week 4, and the spike-specific T-cell responses in HCs were significantly stronger than those in PLWH 4 weeks after the third vaccination. In the AIM assay, spike-specific CD4 + T-cell responses peaked in both PLWH and HCs in week 12. Additionally, significantly higher spike-specific CD8 + T-cell responses were induced in PLWH than in HCs in week 12. In PLWH, the release of the cytokines interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-α), and IL-22 by peripheral blood mononuclear cells (PBMCs) that were stimulated with spike peptides increased in week 12. In addition, the levels of IL-4 and IL-5 were higher in PLWH than in HCs in week 12. Interestingly, the magnitude of SARS-CoV-2-specific T-cell responses in PLWH was negatively associated with the extent of CD8 + T-cell activation and exhaustion. In addition, positive correlations were observed between the magnitude of spike-specific T-cell responses (determined by measuring IFN-γ release by ELISpot) and the amounts of IL-4, IL-5, IL-2 and IL-17F. Conclusions::Our findings suggested that SARS-CoV-2-specific T-cell responses could be enhanced by the booster dose of inactivated COVID-19 vaccines and further illustrate the importance of additional vaccination for PLWH.

ObjectiveTo determine the influence of COVID-19 prevention and control on the epidemic characteristics and dynamics of notifiable infectious diseases in the first quarters， Zhejiang Province， and to explore more effective countermeasures against infectious diseases. MethodsDescriptive epidemiology was conducted to determine the change in notifiable infectious diseases during the prevention and control of COVID-19 in Zhejiang Province by retrieving the data of notifiable infectious diseases from 2017 to 2022 in the Chinese information system for disease control and prevention. Cumulative reported new cases of notifiable infectious diseases in the first quarters of 2017‒2019 were compared with that of 2020‒2022. ResultsA total of 546 753 cases of notifiable infectious diseases were newly reported in the first quarters of 2017‒2019， with an average incidence of 321.92/10⁵. In contrast， a total of 509 908 cases of notifiable infectious diseases were newly reported in the first quarters of 2020‒2022， during which the COVID-19 epidemic occurred， with an average incidence of 270.39/10⁵. The incidence in 2020‒2022 significantly declined by 51.53/10⁵， compared with that in 2017‒2019 （χ²=8 072.06， P<0.001）. In the first quarters of 2020‒2022， the average incidence of zoonotic diseases and vector-borne diseases decreased by more than 50%. In addition， the incidence of respiratory， enteric， blood-borne， and sexually transmitted diseases declined to certain degree. ConclusionThe decline in the newly reported cases of non-COVID-19 notifiable infectious diseases in the first quarters of 2020‒2022 indicates that the countermeasures against COVID-19 epidemic， such as multi-disease co-prevention， multi-sectoral collaboration， societal mobilization and personal hygiene and protection， may also decrease the incidence of multiple infectious diseases. It suggests the countermeasures are effective， which would provide evidence for routine prevention and control of infectious diseases in future.

Corona virus disease 2019 （COVID-19） has led to more than 500 million cases， including 6.19 million deaths. Mutated strains of SARS-CoV-2 （alpha， beta， gamma， delta and omicron） have successively emerged. Currently the omicron variant is gradually replacing the delta variant， driving a surge of cases in most countries including China， posing a great challenge to the prevention and treatment of COVID-19. In the face of the severe pandemic situation， a large number of non-medical volunteers are fighting on the front lines of the war against the pandemic and are vulnerable to exposure due to lack of professional knowledge. This paper introduces the routine skills training of volunteers and emergency response， aiming to provide behavioral guidance for non-medical volunteers in designated isolation facilities.

Upper respiratory tract is directly connected with the external environment， and its natural immune system is the first line of defense against pathogens. In antiviral infection， interferon （IFN） is the main component of the antiviral natural immune system and IFN-λ is a newly discovered immune effector molecule that is mainly produced in the mucosal barrier. IFN-λ exerts a biological role through Janus kinase （JAK） and signal transducer and activator of transcription （STAT） signaling pathway， and plays an important part in regulating innate and acquired immunity of respiratory mucosa. IFN-λ principally expresses on the mucosal barrier with a long-lasting antiviral impact and controls immune-inflammatory damage， which is becoming a new focus of antiviral immunity research in the upper respiratory tract， especially in fighting against 2019 novel coronavirus diseases （COVID-19）. Thus， we summarize the research progress of IFN-λ antiviral immunity in the upper respiratory tract to provide new insight in the prevention and treatment of viral infection in the upper respiratory tract.

Hilar cholangiocarcinoma is the most common extrahepatic malignant tumor in clinic, accounting for about 50%-60% of bile duct tumors. Currently, radical surgical resection is regarded as the best treatment for early hepatic hilar cholangiocarcinoma, but, it’s early lesions have no obvious clinical symptoms. Most patients are already in the advanced stage of the disease when they are admitted to hospital. The surgical resection rate is low, and the 5-year survival rate does not exceed 25%. Locally advanced hepatic hilar cholangiocarcinoma are treated with neoadjuvant chemoradiation therapy followed by surgery, it can prolong survival time of the patient. At the same time, the recurrence rate after surgery can reach 50%-70%, and the cancer easily invades microvessels, lymphatic vessels, peripheral nerves and liver, which is one of the most challenging problems in the field of biliary surgery. Therefore, early diagnosis, perioperative management, adequate and accurate preoperative staging assessment, intraoperative standardized resection, postoperative adjuvant therapy of hilar cholangiocarcinoma, it is of great significance to improve the rate of radical surgical resection and prolong the survival time of patients.

Objective:To explore the influence of short-time pasteurization (62.5±0.5℃ for 5 s) on the main bioactive components and immune cells in human breast milk.Methods:Fresh breast milk was collected from 53 women whose premature infants were admitted to the neonatal intensive care unit of the Children's Hospital of Fudan University from May 2020 to October 2020. Each sample (20 ml) was divided into unsterilized, Holder pasteurized (62.5 ℃ for 30 min), or short-time pasteurized groups. The concentration of secretory immunoglobulin A (sIgA), lactoferrin (LTF), lysozyme (LZM), and insulin-like growth factor binding protein-3 (IGFBP-3) in breastmilk whey were detected by enzyme-linked immunosorbent assay and the number of viable immune cells (leukocytes, monocytes, T cells, and B cells) in breastmilk by flow cytometry.Results:(1) A total of 87 breast milk samples were collected. The levels of sIgA, LTF, and LZM were the highest in the unsterilized group, followed by the short-time and Holder pasteurized group [0.42 mg/ml (0.33-0.65 mg/ml) vs 0.40 mg/ml (0.28-0.62 mg/ml) and 0.25 mg/ml (0.17-0.37 mg/ml); (3.57±1.06) vs (3.53±1.11) and (0.85±0.58) mg/ml; 128.60 μg/ml (77.18-203.00 μg/ml) vs 121.70 μg/ml (68.66-188.20 μg/ml) and 83.40 μg/ml (47.40-151.40 μg/ml); all P<0.05]. There was no significant difference in the level of IGFBP-3 among the groups. The median retention rates of sIgA, LTF, and LZM in the Holder pasteurized group were all lower than those in the short-time pasteurized group [55.87% (46.01%-71.41%) vs 96.93% (83.03%-115.90%); 21.72% (12.54%-29.42%) vs 97.88% (88.98%-104.30%); 69.26% (49.42%-89.08%) vs 93.80% (74.85%-110.20%); all P<0.05]. No significant difference in the level of preserved IGFBP-3 was observed between the three groups ( P>0.05). (2) The number of viable leukocytes, monocytes, T cells, and B cells in the Holder pasteurized group were lower than those in the unsterilized group [leukocytes: 185.50 (87.00-356.50) vs 1 271.00 (540.50-2 283.00); monocytes: 12.00 (6.00-16.75) vs 266.00 (137.30-518.80); T cells: 1.00 (0.00-2.00) vs 47.50 (28.50-116.00); B cells: 1.00 (0.00-1.75) vs 21.00(10.00-41.50); all P<0.05]. The percentage of viable leukocyte to the total leukocyte and the viable monocytes, T cells, and B cells to the viable leukocytes were lower in the Holder pasteurized group than those in the unsterilized group [24.80%(16.00%-36.80%) vs 74.20%(63.55%-86.45%); 5.91%(4.09%-8.77%) vs 21.90%(17.40%-29.30%); 0.31%(0.00%-1.31%) vs 4.00%(2.69%-6.43%); 0.30%(0.00%-0.86%) vs 1.27%(0.57%-2.85%); all P<0.05]. A similar trend was observed between short-time pasteurization and unsterilized groups (all P<0.05). (3) The percentages of viable monocytes, T cells, and B cells in their subsets were lower in both Holder and short-time pasteurized groups than those in the unsterilized group [2.94%(1.33%-7.14%) vs 9.72%(5.77%-16.00%) and 52.60%(31.35%-68.75%); 0.00%(0.00%-1.61%) vs 0.49%(0.00%-2.53%) and 28.10%(10.55%-57.00%); 0.00%(0.00%-0.83%) vs 0.24%(0.00%-2.47%) and 13.80%(3.27%-41.00%); all P<0.05].The number and percentage of viable leukocytes in total leukocytes and viable monocytes in total monocytes [leukocytes: 279.50(116.80-548.50), 32.20%(20.70%-45.75%); monocytes: 32.00(21.00- 83.75),15.60%(11.10%-19.15%)] were higher than those in the pasteurized group (all P>0.05). The short-time pasteurized group was noted only for a higher percentage of the viable monocytes to viable leukocytes than the Holder pasteurized group (all P<0.05). Conclusions:Compared with the Holder pasteurization, sIgA, LTF, LZM level, and monocyte activity in breast milk can be better preserved by short-time pasteurization.

Objective:To investigate whether the bladder neck descent and the area of levator ani hiatus obtained on the maximum Valsalva were consistent at the lithotomy position and separating-legs-holding-knees position.Methods:Forty four subjects who underwent perineal pelvic floor four-dimensional ultrasound at 6-8 weeks postpartum in Third Hospital of Longgang Shenzhen from November 2019 to January 2020 were enrolled in this study. All subjects were able to effectively complete the maximum Valsalva maneuver both at the lithotomy position and the separating-legs-holding-knees position. The bladder neck descent and hiatus area of the levator Ani on the maximum Valsalva at these two positions were measured and compared.Results:The bladder neck descent at the separating-legs-holding-knees position was higher than that at the lithotomy position [(41.06±8.25)mm vs (39.96±8.10)mm; t=-2.965, P=0.005], and the hiatus area of levator Ani at the separating-legs-holding-knees position was larger than that at the lithotomy position[(27.55±4.90)cm 2 vs (26.32±4.76)cm 2; t=-3.820, P<0.001]. Conclusions:The bladder neck descent and the area of levator Ani hiatus on the maximum Valsalva at separating-legs- holding-knees position are larger than those on the maximum Valsalva at the lithotomy position.

Objective To investigate the changes in neutrophil immunophenotypes in neonates with late-onset sepsis and their clinical significance.Methods A total of 42 neonates with late-onset sepsis were enrolled prospectively as sepsis group from Children's Hospital of Fudan University from January 2015 to October 2016,which included 26 preterm infants and 16 term infants.Another 33 neonates without infectious diseases consisting of 20 preterm infants and 13 term infants were selected as control group.According to the severity of sepsis,neonates in the sepsis group were further divided into severe (n=11)and mild sepsis (n=31) subgroups.Expression of CD16 and CD62L on neutrophils was measured by flow cytometry to determine the distribution of neutrophil subsets in neonatal peripheral blood.Differences in the distribution of neutrophil subsets between the two groups and two subgroups were compared using Wilcoxon rank-sum test.Multivariate logistic regression analysis was used to investigate the relationship between neutrophil subsets and the severity of neonatal sepsis.Receiver operating characteristic (ROC) curves were used to assess the diagnostic value of neutrophil subsets for the severity of sepsis.Results Four neutrophil subsets were identified in neonatal peripheral blood,including early-stage neutrophils (CD16-/CD62L-),immature neutrophils (CD16-/CD62L+),mature neutrophils (CD16+/CD62L+) and activated neutrophils (CD16+/CD62L-).Activated [preterm:61.1％ (20.2％-79.4％),term:47.6％ (15.2％-70.1％)] and mature neutrophils [preterm:35.7％ (19.9％-75.8％),term:52.0％ (25.6％-82.8％)] were the dominant subsets in the control group.In preterm infants,the proportion of early-stage [3.5％ (1.7％-9.4％) vs 1.9％ (0.6％-4.0％),Z=-2.501,P=0.012] and immature neutrophils [6.3％ (0.7％-45.5％) vs 0.4％ (0.3％-0.7％),Z=-3.878,P＜0.001] were higher,but that of activated neutrophils [8.3％ (2.3％-49.2％) vs 61.1％ (20.2％-79.4％),Z=2.991,P=0.002] were lower in the sepsis group than those in the control group;same differences were found in the absolute counts of each neutrophil subsets.Among term infants,more immature neutrophils were found in the sepsis group than those in the control group [49 (18-200) vs 13 (5-36)/μl,Z=-2.193,P=0.028].The proportion and the absolute counts of early-stage and immature neutrophils in the severe sepsis subgroup were all higher than those in the mild cases [early-stage neutrophils:5.8％ (3.4％-17.8％) vs 3.0％ (1.4％-7.3％),304 (137-1478) vs 158 (53-321)/μl;immature neutrophils:23.0％ (6.3％-47.0％) vs 0.9％ (0.5％-6.8％),1003 (487-2818) vs 85 (18-275)/μl;all P＜0.05].Multivariate logistic regression analysis showed that the proportion of early-stage neutrophils was associated with the severity of sepsis (OR=1.2,95％CI:1.0-1.4,P=0.012).In addition,the diagnostic value of the proportion of early-stage neutrophils for severe sepsis was the highest when the cut-off value was 3.3％,with the area under the ROC curve was 0.7 (95％CI:0.6-0.9),sensitivity of 81.8％ (95％CI:48.2％-97.7％) and specificity of 62.3％ (95％CI:42.2％-78.2％).Conclusions There are four neutrophil subsets in the peripheral blood of neonates and autoactivation of neutrophils may exist.With the onset of sepsis,neutrophil subsets react differently between preterm and term infants.The proportion of early-stage neutrophils may be correlated with the severity of neonatal sepsis,which may have a predictive value for severe sepsis.

BACKGROUND: Gene therapy and cell transplantation therapy, especially their combined use, have certain therapeutic effects on retinitis pigmentosa. However, little attention has been paid to the combination of gene therapy and cell transplantation in the treatment of retinitis pigmentosa.OBJECTIVE: To investigate the protective effect of adipose-derived mesenchymal stem cells (ADMSCs) modified by growth-associated protein-43 (GAP-43) gene on the retina of rats with retinitis pigmentosa. METHODS: ADMSCs from rats with retinitis pigmentosa were isolated and cultured, and then transfected with GAP-43 lentiviral vector. Sixty retinitis pigmentosa rats were randomized into experimental group (subretinal injection of GAP-43-transfected ADMSCs), control group (subretinal injection of ADMSCs) and sham operation group (PBS injection). The expressions of GAP-43 protein, Rho protein and GS protein in the retina of rats were determined by western blot. The outer nuclear layer thickness of the retina was determined by hematoxylin-eosin staining. RESULTS AND CONCLUSION: After transfected cell transplantation, the expression of GAP-43 protein in the retina was gradually increased with time and showed significant differences at different time post-transplantation (P < 0.05).The expression levels of Rho protein and GS protein in the retina of experimental and control groups were higher than that in the sham operation group (P < 0.05). The expression of Rho protein in the retina of the experimental group was higher than that in the control group (P < 0.05). The expression of GS protein in the retina of the experimental group was lower than that in the control group (P < 0.05). The thickness of retinal outer nuclear layer was ranked as follows: the experimental group > the control group > the sham group, and there was significant difference between groups (P < 0.05). These results show that the ADMSCs modified by GAP-43 gene have protective effect on the retina of rats with retinitis pigmentosa.