Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Background::Hepatitis B virus (HBV) infection is the primary cause of hepatocellular carcinoma (HCC) in China. The target population for HCC screening comprises individuals who test positive for hepatitis B surface antigen (HBsAg). However, current data on the prevalence of HBV infection among individuals who are eligible for HCC screening in China are lacking. We aimed to assess the seroepidemiology of HBV infection among Chinese individuals eligible for HCC screening to provide the latest evidence for appropriate HCC screening strategies in China.Methods::Questionnaires including information of sex, age, ethnicity, marital status, educational level, source of drinking water, as well as smoking and alcohol consumption history and serum samples were collected from females aged 45–64 years and males aged 35–64 years in 21 counties from 4 provinces in eastern and central China between 2015 and 2023. Enzyme-linked immunosorbent assay methods were used to detect the serum HBV marker HBsAg.Results::A total of 603,082 individuals were enrolled, and serum samples were collected for analysis from January 1, 2015 to December 31, 2023. The prevalence of HBsAg positive in the study population was 5.23% (31,528/603,082). The prevalence of HBsAg positive was greater in males than in females (5.60% [17,660/315,183] vs 4.82% [13,868/287,899], χ 2 = 187.52, P <0.0001). The elderly participants exhibited a greater prevalence of HBV infection than younger participants (χ 2 = 41.73, P <0.0001). Birth cohort analysis revealed an overall downward trend in HBV prevalence for both males and females. Individuals born in more recent cohorts exhibited a lower prevalence of HBV infection as compared to those born earlier. Conclusions::The current prevalence of HBV infection remains above 5% in populations eligible for HCC screening in China.

Background::Liver cancer remains the sixth most commonly diagnosed cancer and the third leading cause of cancer-related deaths worldwide, causing a heavy burden globally. An updated assessment of the global epidemiology of the liver cancer burden that addresses geographical disparities is necessary to better understand and promote healthcare delivery.Methods::Data were extracted from the GLOBOCAN 2022 database, including the number, crude, and age-standardized rates of incidence and mortality at the global, country, continent, and human development index (HDI) regional levels. Age-standardized rates (incidence and mortality) per 100,000 person-years were adjusted based on the Segi-Doll World standard population. The mortality-to-incidence ratios (MIR) for each region and country were calculated. The HDI and gross national income (GNI) for 2022 were obtained, and a Pearson correlation analysis was conducted with the incidence, mortality, and MIR.Results::In 2022, approximately 866,136 new liver cancer cases and 758,725 related deaths were recorded worldwide, with a global MIR of 0.86. Males had a disproportionately higher burden than females across all levels, and the highest burden was observed in the elderly population. Geographically, the regions with the highest incidence rates included Micronesia, Eastern Asia, and Northern Africa, and the regions with the highest mortality rates included Northern Africa, Southeastern Asia, Eastern Asia, and Micronesia. Notably, Mongolia had a strikingly high burden compared to other countries. The highest MIR was observed in North America and the lowest in Africa. Negative associations of HDI and GNI with liver cancer mortality and MIR were identified, irrespective of sex.Conclusions::The current liver cancer burden underscores the presence of remarkable geographic heterogeneity, which is particularly evident across countries with varying HDI levels, highlighting the urgent need to prioritize health accessibility and availability to achieve health inequities.

Abstract： Objective　To establish a screening-diagnosis-evaluation system of neonatal congenital heart disease (CHD) suitable for Hainan Province based on the application of percutaneous blood oxygen saturation measurement and cardiac auscultation screening, and to provide a basis for further promotion of the evaluation effect. Methods　Screening agencies (all midwifery institutions) used the "double index method" to screen newborns for congenital heart disease. Those who were screened positive for any index were transferred to diagnostic institutions (the people's hospitals of all cities and counties and some tertiary hospitals) for echocardiography examination within 1 week. Those with abnormal results were transferred to the treatment institution (two tertiary hospitals) for reconfirmation and evaluation. All data were managed online. Results　In this study, 96 913 newborns born in midwifery institutions in 19 cities and counties were screened, with a response rate of 99.44%. The ratio of male infants (50 836) to female infants (46 077) was 1.10∶1. A total of 2 284 positive patients were screened by dual index method, and ultrasound diagnosis was performed, with the rate of cardiac ultrasonography of 98.07% (2 240/2 284) and a high response rate. A total of 238 cases of congenital heart disease were diagnosed (incidence rate of 0.25%), with the top three main types were ventricular septal defect, atrial septal defect, and patent ductus arteriosus. The sensitivity (88.24%) and Youden index (0.86) of dual-indicator screening for CHD detection were significantly higher than the other separate indicators. Through Kappa consistency test analysis, the consistency of dual-indicator screening with simple heart murmur screening results was excellent, with a Kappa coefficient value of 0.835 (>0.75); the consistency of dual-indicator screening and simple POX screening results was good, with a Kappa coefficient value of 0.429 (between 0.40-0.75), and the differences were statistically significant (P<0.001). By the end of the study, 136 children had undergone open-chest/occlusion surgery, with good postoperative recovery, and the rest were followed up. Conclusions　The neonatal CHD screening-diagnosis-assessment technology system established in this study, with close connections between various blocks, high screening response rate and echocardiography examination rate, is beneficial for the early diagnosis and treatment of CHD children, and has certain application value in institutions at all levels in Hainan Province, and is worth further promotion.

ObjectiveTo identify causal factors of a case of severe Chlamydia psittaci pneumonia in Yangpu District and provide a scientific basis for effective prevention and control. MethodsBasic information and epidemiological data of the patient were collected through telephone interviews and field epidemiological surveys. Specimens from the patient， close contacts and the environment were collected for pathogen detection. Metagenomics next-generation sequencing （mNGS） was used to identify unknown pathogens. ResultsA 65-year-old male patient with a history of hypertension and diabetes was admitted to the hospital with symptoms of fatigue， poor appetite for a week， fever and cough for four days. A chest computer tomography （CT） scan showed scattered inflammation in the left lung with infiltration of multiple lobes. Blood gas analysis showed type I respiratory failure. The results of mNGS on the bronchoalveolar lavage fluid of the patient indicated that he was infected with Chlamydia psittaci. Epidemiological investigation showed a clear history of avian exposure， with an incubation period of 30 days. ConclusionThis serious pneumonia is a zoonotic disease caused by Chlamydia psittaci. A clear history of avian exposure and the use of mNGS technology can help in the timely diagnosis of this disease.

Objective: To investigate the awareness of human papillomavirus (HPV) vaccine knowledge among women in Hainan Province and the willingness to receive HPV vaccination among women and their daughters, so as to provide insights into HPV vaccine promotion and cervical cancer control. Methods: Women aged 20 to 64 years who participated in the cervical cancer screening program in Hainan Province from July 2021 to February 2022 were enrolled, and participants' demographics, awareness of knowledge about HPV vaccine, and willingness to receive HPV vaccination among themselves and their daughters using a questionnaire survey. The factors affecting willingness to receive HPV vaccination were identified using a multivariable logistic regression model. Results: A total of 4 300 questionnaires were allocated and 4 023 valid questionnaires were recovered, with an effective recovery rate of 93.56%. The respondents had a mean age of (35.52±6.48) years, and their daughters had a mean age of (8.93±4.83) years. There were 826 participants knowing HPV vaccine-related knowledge, with an awareness rate of 20.35%. The proportions of willingness to receive HPV vaccination were 75.07% among respondents and 71.36% among their daughters. Multivariable logistic regression analysis identified age (20 to <30 years, OR=1.474, 95%CI: 1.167-1.862; 30 to <40 years, OR=1.455, 95%CI: 1.199-1.765), urban-registered residence (OR=1.251, 95%CI: 1.016-1.541), monthly household income (3 000 to <5 000 Yuan, OR=1.568, 95%CI: 1.314-1.873; 5 000 Yuan and more, OR=1.231, 95%CI: 1.014-1.496), affordable fees for a single dose of HPV vaccination (100 to <500 Yuan, OR=1.358, 95%CI: 1.159-1.590; 500 Yuan and higher, OR=1.609, 95%CI: 1.188-2.180), and awareness of HPV vaccine-related knowledge (OR=4.473, 95%CI: 3.416-5.855) as factors affecting respondents' willingness to receive HPV vaccination, and participation in New Rural Cooperative Medical Insurance (OR=1.652, 95%CI: 1.087-2.511), affordable fees for a single dose of HPV vaccination (100 to <500 Yuan, OR=1.905, 95%CI: 1.633-2.221; 500 Yuan and higher, OR=1.683, 95%CI: 1.275-2.223), awareness of HPV vaccine-related knowledge (OR=3.625, 95%CI: 2.847-4.615), daughter's active request for HPV vaccination (OR=1.695, 95%CI: 1.293-2.222), and frequency of sexual health education for daughters (frequently, OR=2.142, 95%CI: 1.789-2.564; occasionally, OR=1.707, 95%CI: 1.416-2.059) as factors affecting the willingness to receive HPV vaccination among respondents' daughters. Conclusions The awareness of HPV vaccine-related knowledge is low among women aged 20 to 64 years in Hainan Province, and the women's and their daughters' willingness to receive HPV vaccination is associated with age, household registration, economic level, price of HPV vaccine and awareness of HPV vaccine-related knowledge.

Abstract: Objective　To evaluate the free thalassaemia screening programme for preconception and pregnancy in Hainan Province, and to provide a theoretical basis for optimizing the screening process for thalassaemia. Methods From November 2020 to July 2021, a survey was conducted on 10 396 adults with Hainan household registration who participated in the Epidemiological Survey of Thalassemia in Hainan Residents in 19 cities and counties of Hainan Province. All of them underwent routine blood tests, haemoglobin electrophoresis tests and genetic tests for thalassaemia. The optimal diagnostic cut-off values for mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), and haemoglobin adult type 2 (HbA2) were determined using screening test indexes such as receiver operating characteristic curve and sensitivity. The diagnostic effectiveness of different primary screening programs for thalassemia gene carriers was evaluated. Results Using the existing MCV single-indicator thalassemia primary screening protocol in Hainan Province, where individuals with MCV<82 fL undergo thalassemia gene testing, resulted in a high missed diagnosis rate (34.06%) and low sensitivity (65.94%). The optimal cut-off values for MCV screening for alpha-and beta-thalassaemia were 84.45 fL and 79.05 fL, respectively; the optimal cut-off values for MCH screening for alpha-and beta-thalassaemia were 27.95 pg and 25.15 pg, respectively. The optimal cut-off value for HbA2 screening for alpha-thalassaemia was less than 2.55% and greater than 3.35% for beta-thalassaemia. The "combined HbA2 or MCH or MCV screening protocol" with the cut-off values recommended in this study had a better performance in primary screening for thalassemia, with the highest sensitivity (92.96%) and negative predictive value (92.67%) and the lowest underdiagnosis rate (7.04%), statistically significant differences compared with the existing protocol (P<0.05). Conclusions The current process of screening for thalassemia in Hainan Province may lead to missed diagnoses. The combined use of MCV, MCH and HbA2 for thalassemia screening, adopting locally suitable cutoff values for primary screening indicators, can improve the incidence of missed reporting of thalassemia and enhance diagnostic effectiveness.

Objective:To investigate the change trend of etiological burden of disease of liver cancer in the Chinese population from 1990 to 2019.Methods:The descriptive epidemiologic method was conducted. Based on the Global Burden of Disease data from the Institute for Health Metrics and Evaluation at the University of Washington, the data related to liver cancer burden caused by hepatitis B virus (HBV) infection, hepatitis C virus (HCV) infection, alcohol, nonalcoholic steatohepatitis (NASH) and other factors, including number of new cases, the crude incidence rate, age-specific incidence rate, number of deaths, crude mortality rate and age-specific mortality rate, in the Chinese population from 1990 to 2019 were collected. The age-standardized rate was calculated based on the world standardized population structure in 2019 from the Global Burden of Disease data. Observation indicators: (1) the incidence of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019; (2) the mortality of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019; (3) the change trend of age-specific incidence rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019; (4) the age-specific mortality rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. Count data were expressed as absolute numbers, percentages and ratio. Based on the junction point regression model, the Joinpoint software (V.4.9.1.0) was used to calculate the annual percentage change, average annual percentage change (AAPC) and 95% confidence intervals ( CI) of age-specific incidence rate and age-specific mortality rate of liver cancer caused by different etiologies. Results:(1) The incidence of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the number of new cases of liver cancer in Chinese population decreased from 236 825 to 210 462, and the crude incidence rate decreased from 20.01/100,000 to 14.80/100,000. The new cases of liver cancer caused by HBV infection, HCV infection and other factors showed a downward trend, and the absolute change rates were ?14.76%, ?3.98% and ?26.67%, respectively. The new cases of liver cancer caused by alcohol and NASH showed a increase trend, and the absolute change rates were 9.31% and 13.91%, respectively. (2) The mortality of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the number of deaths of liver cancer in Chinese population decreased from 232 449 to 187 700, and the crude mortality rate decreased from 19.64/100,000 to 13.20/100,000. The number of deaths of liver cancer caused by HBV infection, HCV infection and other factors showed a down-ward trend, and the absolute change rates were ?23.34%, ?10.99% and ?33.75%, respectively. The number of deaths of liver cancer caused by alcohol showed a slow downward trend, and the absolute change rate was ?0.51%. The number of deaths of liver cancer caused by NASH showed a increase trend, and the absolute change rate was 6.03%. (3) The change trend of age-specific incidence rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the AAPC of age-specific incidence rate of liver cancer caused by HBV infection, HCV infection, alcohol, NASH and other factors was ?3.61%(95% CI as ?4.10% to ?3.11%), ?3.57%(95% CI as ?3.99% to ?3.14%), ?2.79%(95% CI as ?3.24% to ?2.33%), ?2.65%(95% CI as ?3.09% to ?2.21%) and ?3.62%(95% CI as ?4.05% to ?3.19%), respectively. (4) The age-specific mortality rate of liver cancer caused by different etiologies in the Chinese population from 1990 to 2019. From 1990 to 2019, the AAPC of age-specific mortality rate of liver cancer caused by HBV infection, HCV infection, alcohol, NASH and other factors was ?3.92%(95% CI as ?4.42% to ?3.41%), ?3.90%(95% CI as ?4.45% to ?3.35%), ?3.15%(95% CI as ?3.71% to ?2.58%), ?2.86%(95% CI as ?3.34% to ?2.38%) and ?4.09%(95% CI as ?4.64% to ?3.55%), respectively. Conclusions:From 1990 to 2019, the liver cancer burden of the Chinese population shows an overall downward trend, in which the liver cancer burden caused by HBV and HCV infection decreases the most, but HBV and HCV infection is still the main reason for the heavy burden of liver cancer. The age-specific incidence rate and age-specific mortality rate of liver cancer caused by alcohol and NASH show a downward trend, but the number of new cases of liver cancer caused by alcohol and NASH shows significant growth. The liver cancer burden caused by other factors shows a downward trend.

Abstract: Objective To explore the trend of maternal mortality in Hainan Province during the period of 2003-2022, both in the province and in urban and rural areas, and to forecast the maternal mortality rate for the period 2023-2025. Methods The 2003-2022 data collected from Hainan Province's three-tier network for maternal mortality surveillance was examined using the Chi-square test for trend (CMH) to analyze the province-wide and urban-rural maternal mortality trends. The time series model forecasting using exponential smoothing was used to predict the maternal mortality rate in Hainan Province from 2023 to 2025. Results　From 2003 to 2022, a total of 201 011 616 live births and 463 maternal deaths were reported in Hainan Province, with a maternal mortality rate of 23.03 per 100 000. Over 20 years, the maternal mortality rate in Hainan Province showed a downward trend, with an annual rate of decline of -4.13%. The rate decreased significantly during this period.. From 2003 to 2022, the maternal mortality rate in rural areas of Hainan Province was 25.74/100 000 (373/1 448 943), and it was significantly higher than that in urban areas, 16.04/100 000 (90/561 173). In the first 10 years, the gap between urban and rural areas progressively reduced, but it widened significantly in the last decade, especially after 2017. The maternal mortality rate was significantly lower in urban than rural areas, and the differences had statistical significance. The annual rates of decline in maternal mortality in Hainan Province and in urban and rural areas from 2003 to 2022 were -5.0% and -3.71%, respectively, showing a negative growth with the decrease rate in rural areas lower than urban areas. The maternal mortality rate in Hainan Province showed a fluctuating downward trend, different from the stable trend of national decrease. The mortality rates for direct obstetric causes of death (233 cases) and indirect obstetric causes of death (230 cases) were 11.59/100 000 and 11.44/100 000, respectively. The results of the maternal mortality review showed a predominance of avoidable deaths (315 cases, 68.03%). Brown's cubic exponential smoothing predicted the maternal mortality rate in Hainan Province for 2023-2025 as 9.45/100 000, 8.17/100 000, and 6.89/100 000. Conclusions　The maternal mortality rate in Hainan Province is largely influenced by maternal deaths in rural areas, and maternal health care in rural areas should be emphasized. Measures such as intervening to address the main factors influencing avoidable maternal deaths, strengthening high-risk maternal management, improving the level of critical maternal care, and providing subsidies for critical maternal care can sustainably reduce the maternal mortality rate in Hainan.