Objective To investigatethe relationship between gastroesophageal reflux disease (GERD) symptoms and clinicopathological characteristics, p53 expression, and survival of Chinese patients with esophageal adenocarcinoma. Methods A total of 3882 patients with esophageal adenocarcinoma, 970 matched patients with esophageal squamous cell carcinoma, and 970 matched patients with gastric cardia adenocarcinoma were included to compare the incidence of GERD symptoms. Patients with esophageal adenocarcinoma were divided into two groups according to the presence and absence of GERD symptoms. The differences in clinicopathological characteristics and p53 expression between the two groups were compared by chi-square test, and the differences in survival between the two groups were compared by landmark analysis. Results The incidence of GERD symptoms increased successively in esophageal squamous cell carcinoma, esophageal adenocarcinoma, and gastric cardia adenocarcinoma. In patients with esophageal adenocarcinoma, the proportions of male, less than 65 years old, lower thoracic tumors, tumors without squamous cell carcinoma, poorly differentiation, early tumors (stage 0-I), and p53-positive tumors in the group with GERD symptoms were higher than those in the group without GERD symptoms; moreover, the long-term survival (≥15 years) was better. Conclusion GERD symptom is an important clinical sign of esophageal adenocarcinoma. It is closely related to specific clinicopathological characteristics, p53 overexpression, and good long-term prognosis.

Objective:To understand the epidemiological characteristics of human respiratory syncytial virus (HRSV) among acute respiratory infection (ARI) cases in 16 provinces of China from 2009 to 2023.Methods:The data of this study were collected from the ARI surveillance data from 16 provinces in China from 2009 to 2023, with a total of 28 278 ARI cases included in the study. The clinical specimens from ARI cases were screened for HRSV nucleic acid from 2009 to 2023, and differences in virus detection rates among cases of different age groups, regions, and months were analyzed.Results:A total of 28 278 ARI cases were enrolled from January 2009 to September 2023. The age of the cases ranged from<1 month to 112 years, and the age M ( Q1, Q3) was 3 years (1 year, 9 years). Among them, 3 062 cases were positive for HRSV nucleic acid, with a total detection rate of 10.83%. From 2009 to 2019, the detection rate of HRSV was 9.33%, and the virus was mainly prevalent in winter and spring. During the Corona Virus Disease 2019 (COVID-19) pandemic, the detection rate of HRSV fluctuated between 6.32% and 18.67%. There was no traditional winter epidemic peak of HRSV from the end of 2022 to the beginning of 2023, and an anti-seasonal epidemic of HRSV occurred from April to May 2023. About 87.95% (2 693/3 062) of positive cases were children under 5 years old, and the difference in the detection rate of HRSV among different age groups was statistically significant ( P<0.001), showing a decreasing trend of HRSV detection rate with the increase of age ( P<0.001). Among them, the HRSV detection rate (25.69%) was highest in children under 6 months. Compared with 2009-2019, the ranking of HRSV detection rates in different age groups changed from high to low between 2020 and 2023, with the age M (Q1, Q3) of HRSV positive cases increasing from 1 year (6 months, 3 years) to 2 years (11 months, 3 years). Conclusion:Through 15 years of continuous HRSV surveillance analysis, children under 5 years old, especially infants under 6 months old, are the main high-risk population for HRSV infection. During the COVID-19 pandemic, the prevalence and patterns of HRSV in China have changed.

Objective:To understand the epidemiological characteristics of human respiratory syncytial virus (HRSV) among acute respiratory infection (ARI) cases in 16 provinces of China from 2009 to 2023.Methods:The data of this study were collected from the ARI surveillance data from 16 provinces in China from 2009 to 2023, with a total of 28 278 ARI cases included in the study. The clinical specimens from ARI cases were screened for HRSV nucleic acid from 2009 to 2023, and differences in virus detection rates among cases of different age groups, regions, and months were analyzed.Results:A total of 28 278 ARI cases were enrolled from January 2009 to September 2023. The age of the cases ranged from<1 month to 112 years, and the age M ( Q1, Q3) was 3 years (1 year, 9 years). Among them, 3 062 cases were positive for HRSV nucleic acid, with a total detection rate of 10.83%. From 2009 to 2019, the detection rate of HRSV was 9.33%, and the virus was mainly prevalent in winter and spring. During the Corona Virus Disease 2019 (COVID-19) pandemic, the detection rate of HRSV fluctuated between 6.32% and 18.67%. There was no traditional winter epidemic peak of HRSV from the end of 2022 to the beginning of 2023, and an anti-seasonal epidemic of HRSV occurred from April to May 2023. About 87.95% (2 693/3 062) of positive cases were children under 5 years old, and the difference in the detection rate of HRSV among different age groups was statistically significant ( P<0.001), showing a decreasing trend of HRSV detection rate with the increase of age ( P<0.001). Among them, the HRSV detection rate (25.69%) was highest in children under 6 months. Compared with 2009-2019, the ranking of HRSV detection rates in different age groups changed from high to low between 2020 and 2023, with the age M (Q1, Q3) of HRSV positive cases increasing from 1 year (6 months, 3 years) to 2 years (11 months, 3 years). Conclusion:Through 15 years of continuous HRSV surveillance analysis, children under 5 years old, especially infants under 6 months old, are the main high-risk population for HRSV infection. During the COVID-19 pandemic, the prevalence and patterns of HRSV in China have changed.

Objective To investigate the impact of overweight on the prognosis of patients with acute mild ischemic stroke or moderate-high risk transient ischemic attack (TIA). Methods A total of 366 patients with acute mild ischemic stroke or moderate-high risk TIA who carried CYP2C19 loss-of-function alleles were included in the study. Baseline data were collected at admission, and clinical outcomes were collected within 3 months post-onset (primary outcomes: stroke recurrences within 3 months post-onset; secondary outcomes: composite outcome of stroke recurrence or death within 3 months post-onset, vascular events, and quality of life within 3 months). Kaplan-Meier method was used to plot the cumulative incidence curve of outcomes. Multivariate Cox proportional hazards model and multivariate Logistic regression model were used to evaluate the relationship between overweight and clinical outcomes within 3 months post-onset in patientswith ischemic stroke or TIA. The integrated discrimination improvement index (IDI) and net reclassification improvement index (NRI) were calculated to assess the predictive value of adding body mass index (BMI, whether overweight or not) based on traditional models for predicting the prognosis of acute ischemic stroke or TIA. Results After 3 months of follow-up, 28 patients had stroke recurrence, 1 patient died, and 31 patients had vascular events. The Kaplan-Meier cumulative incidence curve showed that the cumulative incidences of stroke recurrence, stroke recurrence or death, and vascular events after 3 months of onset were lower in overweight patients compared with non-overweight patients (Log-rank P < 0.05). Multivariate analysis showed that compared with non-overweight patients, overweight patients had significantly reduced risks of stroke recurrence within 3 months (HR=0.24; 95%CI, 0.08 to 0.71), composite outcome of stroke recurrence or death (HR=0.24; 95%CI, 0.08 to 0.69), and vascular events (HR=0.22; 95%CI, 0.07 to 0.63), and significantly improved quality of life within 3 months (OR=0.39; 95%CI, 0.20 to 0.76). The IDI and NRI calculations showed that compared with traditional models, the new model adding BMI (whether overweight) had significantly improved predictive ability for the prognosis of acute ischemic stroke or TIA. Conclusion Overweight may be a protective factor for the prognosis of TIA patients with acute mild ischemic stroke or moderate-high risk who carry CYP2C19 loss-of-function alleles. Compared with non-overweight patients, overweight patients have reduced risks of stroke recurrence, composite outcome of stroke recurrence or death, and vascular events within 3 months post-onset, and improved quality of life within 3 months post-onset.

Objective:To study characteristics of changes in the demand for emergency medical services during epidemic prevention and control "10 new measures" in Zhejiang province.Methods:The data of 26 emergency centers connected to the provincial integration platform of Zhejiang Province (hereinafter referred to as "provincial platform" ) were retrospectively analyzed, and the data were collected from one week before the implementation of "10 new measures" to the sixth week after implementation (December 1, 2022 to January 18, 2023). The collected information included: the number of 120 calls and ambulance services, the types of disease, age composition of patients, performance of emergency medical services.Results:From the second week of the implementation of "10 new measures" (December 15 to 21, 2022), the number of 120 calls and ambulance services were increased rapidly, and the peak occurred in the third week of implementation (December 21 to 28, 2022). Among the types of diseases, the number and proportion of patients with abnormal symptoms and respiratory diseases increased significantly, reaching the highest peak in the third week (December 21 to 28, 2022) and the fourth week (December 29, 2022 to January 4, 2023) of implementation, respectively. After the second week of implementation, the number of elderly patients aged 71 to 100 increased significantly, reaching a peak in the fourth week (December 29, 2022 to January 4, 2023), accounting for 60.76% of the total. During the epidemic period, the quality control indicators such as emergency dispatch time, ambulance dispatch time and medical treatment all fluctuated, but the changes were not significant.Conclusions:During the implementation of epidemic prevention and control "10 new measures", there were obvious characteristic changes in the demand for pre-hospital emergency in Zhejiang Province, but the quality of pre-hospital emergency medical was basically stable.

Objective:To explore the efficacy and safety of BCL-2 inhibitor-based treatment in patients with relapsed/refractory t (11; 14) primary systemic light chain amyloidosis.Methods:This was a retrospective case series study. Ten patients with relapsed/refractory t(11;14) primary systemic light chain amyloidosis who had all received treatment with a combination regimen including the BCL-2 inhibitor venetoclax from January 2018 to November 2022 at the Hematology Department of Peking University People′s Hospital were included. Adverse events, and hematological and organ responses were evaluated.Results:The median age of the ten enrolled patients was 59 (range 41-78) years, and the male to female ratio was 8∶2. Except for one patient, a very good partial or better response was achieved in 8/9 patients and one patient obtained a partial response. The overall response rate was 100%. The median time to achieve a hematological response was 60 (range 24-236) days. At least one organ response was observed in 7/9 patients. With a median follow-up of 18 months, one patient experienced hematological progression and one patient died. Grade 3 adverse events included lymphocytopenia (3 cases), anemia (1 case), diarrhea (1 case), and appendicitis (1 case). One patient died of pulmonary fungal infection two months after completion of treatment, which was not excluded as being treatment related.Conclusion:A combination regimen including BCL-2 inhibitors in patients with relapsed/refractory t(11;14) primary systemic light chain amyloidosis is a potentially safe and effective treatment option that warrants further investigation.

ObjectiveTo study the effect-enhancing and toxicity-reducing activities of astragalus polysaccharide injection （APS） on U14 cervical cancer in model mice receiving X-ray treatment. MethodU14 mouse cervical cancer cells were cultured in vitro and injected into the right forelimb armpit of Kunming mice for constructing a subcutaneous tumor-bearing model of cervical cancer. The tumor-bearing mice were randomly divided into the model group， X-ray intervention（IR， 6 Gy） group， APS （10 mL·kg^-1·d^-1） group， and IR + APS group. Following the observation of the state， body mass， and food intake of mice in each group， the volume of the tumor was measured. The tumor cell cycle and apoptosis were determined by flow cytometry. The protein and mRNA expression levels of apoptosis-related proteins p53， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cleaved cysteine-dependent aspartate-directed protease-3 （Caspase-3） in tumor tissues were assayed by Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultThe comparison with the model group showed that mice in the IR group had poor mental status and reduced mobility. The IR group and IR + APS group exhibited reduced food intake and body mass since the 8^th d （P<0.05， P<0.01） and narrowed tumor volume since the 9^th d （P<0.01）. In the IR group， the proportion of cells in the G₁ phase was increased， while the proportion of those in the S phase was decreased （P<0.01）. In the IR + APS group， the proportion of cells in the G₁ phase rose， whereas the proportion of those in the G₂ and S phases cells declined （P<0.05， P<0.01）. The apoptotic rates in both the IR group and IR + APS group were elevated significantly （P<0.01）. Compared with the model group， the IR group and IR + APS group displayed up-regulated cleaved Caspase-3 and Bax protein and mRNA expression in tumor tissues， but down-regulated Bcl-2 and p53 protein and mRNA expression （P<0.05， P<0.01）. Compared with the IR group， the mice in the IR + APS group had better mobility and hair， normal body mass， and increased food intake （P<0.05）. The tumor volume in the IR + APS group was reduced （P<0.05）. The proportion of cells in the G₂ phase was reduced， but the proportion of those in the S phase was raised （P<0.05）. The apoptosis rate was increased （P<0.05）. The apoptosis-related protein Bax protein expression in the tumor tissue was up-regulated， while the protein expression levels of Bcl-2 and p53 were down-regulated （P<0.05， P<0.01）. ConclusionAPS maintains the life state of U14 cervical cancer model mice treated with X-ray and promotes tumor cell apoptosis， thus enhancing the efficiency and reducing toxicity.

Objective:To understand the genetic characteristics of human adenovirus (HAdV) in severe acute respiratory infection (SARI) cases in Luohe city, Henan province, China.Methods:After viral isolation of HAdV-positive specimens identified in SARI cases from October 2017 to February 2019, the Loop2 region of the Hexon gene was amplified and determined to initially identify the virus type. Then, based on the preliminary screening result, the full length of the sequences of the three target genes (Penton base, Hexon and Fiber) of the viral strains were amplified using specific primers for each HAdV type, and phylogenetic and sequence homology analyses were performed with the prototype strains and the representative strains of the corresponding types at home and abroad to identify the types of viral strains and understand their genetic characteristics.Results:A total of 18 viral isolates were obtained from 27 HAdV-positive throat swab specimens from 783 SARI cases in Luohe city, Henan province, and the molecular typing result showed that these strains belonged to species B (HAdV-3, HAdV-7 and HAdV-55), species C (HAdV-1, P1H2F2, Px1/Ps3H5F5, P89H5F5 and HAdV-6) and species E (HAdV-4). Among them, the highest positive detection rate was found for species C HAdV-1 isolates (33.3%), followed by species B HAdV-3 (22.2%) and species C P1H2F2 (11.1%). The four HAdV strains in this study (HAdV-3, HAdV-4, HAdV-7 and HAdV-55) were characterized by significantly conserved and stable in time and space; while three patterns of genetic recombination (P1H2F2, Px1/Ps3H5F5 and P89H5F5) were identified for HAdV-C strains in this study, and their potential public health significance needed to be confirmed by further studies.Conclusions:The HAdV infection of SARI cases in Luohe city, Henan province during 2017-2019 was dominated by species C, followed by species B and species E. These data provided a scientific basis for the prevention and control of local adenovirus-associated infectious diseases.

Objective To evaluate the inhibitory effect of fenretinide-loaded liposomes(4-HPR-L) on subcutaneous transplanted malignant melanomas in nude mice.Methods A film-ultrasonic dispersion method was used to prepare 4-HPR-L.BALB/c nude mice were subcutaneously inoculated with A375 melanoma cells in the right axillary fossae to establish malignant melanoma-bearing nude mouse models.Ten nude mouse models were randomly and equally divided into 2 groups to be injected with near-infrared fluorescent cell membrane label (DiR) solution or DiR liposomes (DiR-L)at the same concentration in the caudal vein,and a live imaging system was used to observe the distribution of DiR or DiR-L in nude mice at 6,12,24 hours after the injection.Another 30 nude mice were randomly and equally divided into 3 groups to be injected with 5％ (mass fraction) glucose solution at a single-dose of 0.2 ml (control group),25 mg/kg 4-HPR solution (4-HPR group)and 25 mg/kg 4-HPR-L solution (4-HPR-L group) respectively on days 8,10,12,14,16,18,20 and 22 after the inoculation with A375 cells.The mouse body weight and tumor volume were dynamically monitored in the above groups after the injection,and the survival situation was observed.The nude mice were sacrificed on day 2 after the final injection,and the heart,liver,spleen,lung,kidney and tumor tissues were resected.These tissues were subjected to hematoxylin-eosin staining and immunohistochemical staining to observe the metastasis of melanoma in mice,and terminal deoxyribonucleotidyl transferase-mediated nick end labelling was performed to detect the apoptosis in tumor cells.One-way analysis of variance and independent-sample t test were used to analyze measurement data.Results The live imaging system showed that DiR-L could be retained in melanoma for a long time,and strong fluorescence of DiR-L could be still observed in the tumors at 24 hours after injections.Quantitative fluorescence analysis revealed that the fluorescence intensity of DiR-L (22.85 ± 1.66) was significantly higher than that of DiR in tumor tissues (8.45 ± 0.97,t =12.957,P ＜ 0.01).Compared with the control group and 4-HPR group,the resected tumor weight on day 2 after the final injection was significantly decreased in the 4-HPR-L group (F =27.055,t =4.768,6.640,respectively,both P ＜ 0.05).Hematoxylineosin staining showed that liver metastasis occurred in 2 nude mice in the 4-HPR-L group,but in all the nude mice in the control group and 4-HPR group.All the nude mice in the 4-HPR-L group died within 76 days after inoculation,and the mice in the control group and 4-HPR group all died within 56 and 59 days respectively after inoculation.There were significant differences in the apoptotic index among the control group (12.14‰ ± 1.33‰),4-HPR group (67.17‰± 15.18‰) and 4-HPR-L group (152.73‰ ± 11.27‰;F =167.588,P ＜ 0.05),and the apoptotic index was significantly higher in the 4-HPR-L group than in the control group and 4-HPR group (t =18.162,11.075 respectively,both P ＜ 0.05).Conclusion 4-HPR-L can effectively inhibit the growth of subcutaneous melanoma in nude mice and metastasis of melanoma cells,and prolong the survival duration of nude mice.

Objective To assess the effect of fenretinide (4-HPR) on proliferation,apoptosis and migration of B16F10 and A375 melanoma cells,and to evaluate the effect of liposomes and RGD peptidemodified liposomes on its uptake and therapeutic effects.Methods A film-hydration method was used to prepare 4-HPR liposomes (4-HPRL),which were modified with RGD peptide to prepare RGD-4-HPRL,and the concentration,particle size,electric potential,drug loading capacity and encapsulation efficiency were measured for 4-HPRL and RGD-4-HPRL.In vitro cultured B16F10 and A375 cells were divided into several groups:4-HPR group,4-HPRL group and RGD-4-HPRL group treated with Dulbecco's minimum essential medium (DMEM) containing 4-HPR bulk drug,4-HPRL and RGD-4-HPRL respectively at the same concentration of 4-HPR,and control group treated with culture solution at the same volume.After different durations of treatment,cell counting kit-8 (CCK8) assay was performed to evaluate cellular proliferative activity,annexin V/propidium iodide staining to detect apoptosis,and scratch wound healing assay to evaluate the effect of drug treatment on cell migration ability.Then,4-HPR was replaced by coumarin 6 (C6) to prepare C6 liposomes (C6L) and RGD-C6L,and flow cytometry was conducted to evaluate C6 uptake by B16F10 cells.Statistical analysis was carried out with SPSS22.0 software by one-way analysis of variance (ANOVA) for the comparison among several groups and t test for the comparison between two groups.Results The concentration of 4-HPR in the prepared 4-HPRL solution was over 1 300 mg/L.The encapsulation efficiency and drug loading capacity of 4-HPRL were (95.51 ± 1.22)％ and (7.27 ± 0.11)％ respectively,and those of RGD-4-HPRL were (95.82 ± 0.81)％ and (7.14 ± 0.13)％ respectively.The particle size distribution of 4-HPRL and RGD-4-HPRL was uniform,and their average particle size was below 100 nm.CCK8 assay showed that 4-HPR could markedly inhibit the proliferative activities of B16F10 and A375 cells.The cell proliferation inhibition rate of 4-HPRL was higher than that of 4-HPR at the same concentration of 4-HPR (P ＜ 0.01),and the inhibition rate of RGD-4-HPRL was higher than that of 4-HPRL (P ＜ 0.01 or P ＜ 0.05) and 4-HPR (P ＜ 0.01).As annexin V/propidium iodide apoptosis assay showed,when the concentration of 4-HPR was 10 mg/L,the total apoptosis rates of B16F10 cells in the control group,4-HPR group,4-HPRL group and RGD-HPRL group were (4.44 ± 0.35)％,(28.33 ± 0.66)％,(46.43 ± 0.77)％ and (51.33 ± 0.37)％ respectively.When the concentration of 4-HPR was 20 mg/L,the total apoptosis rates of A375 cells in the above 4 groups were (4.97 ± 0.62)％,(16.68 ± 3.81)％,(32.62 ± 1.24)％ and (44.85 ± 4.92)％ respectively.The apoptosis rates of B16F10 and A375 cells were significantly higher in the 4-HPRL group than in the 4-HPR group (both P ＜ 0.01),and higher in the RGD-4-HPRL group than in the 4-HPRL group (both P ＜ 0.01) and 4-HPR group (both P ＜0.01).Scratch wound healing assay showed that 4-HPR could inhibit scratch healing and migration of B16F10 and A375 cells,and the inhibitory effects of 4-HPRL and RGD-4-HPRL were distinctly superior to those of 4-HPR bulk drug.C6 uptake assay revealed that the fluorescence intensity of C6 in B16F10 cells in the control group,C6 group,C6L group and RGD-C6L group were 2.15 ± 0.28,8.56 ± 0.36,20.48 ± 0.13 and 22.55 ± 0.07 respectively,and there were significant differences between the 4 groups (F =67 194.186,P ＜ 0.01).Additionally,the fluorescence intensity of C6 was significantly higher in the C6L group and RGD-C6L group than in the C6 group (both P ＜ 0.01),and higher in the RGD-C6L group than in the C6L Group (P ＜ 0.01).Conclusions 4-HPR can inhibit the proliferation and migration of A375 and B16F10 cells,and induce their apoptosis.Liposomes and RGD-targeted liposomes can markedly enhance the effect of 4-HPR on melanoma cells.