Background and purpose:In recent years, epigenetics research has become a new direction of cancer research. A large number of results have shown that the abnormal changes of epigenetic modifications have close connection with cancer. Genome-wide epigenetic modifications have become new markers for cancer. This study aimed to investigate the methylation of the promoter ofDBC1 gene in cervical cancer tissues of Uyghur women in Xinjiang, to explore the correlation between the gene methylation and the infection of HPV, and to evaluate whether it can be used as a tool with high sensitivity and specificity for cervical cancer screening.Methods:This study detected the infection of HPV16, 18 in 43 normal cervical tissues, 35 cervical intraepithelial neoplasia tissues and 54 cervical cancer tissues using the polymerase chain reaction (PCR) method. The methylation of the promoter ofDBC1 gene in above-mentioned tissues was detected by the methylation-specific PCR method. The expression ofDBC1 at mRNA level was measured by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) in 10 methylation-negative normal cervical tissues and 10 methylation-positive cervical cancer tissues.Results:In normal cervical tissues, CIN tissues and cervical cancer tissues, the infection ratios of HPV16 were 18.6%, 34.3% and 68.5%, respectively; the infection ratios of HPV18 were 2.3%, 8.6% and 16.7%, respectively; and the methylation ratios ofDBC1 gene were 23.3%, 40.0%, 87.0%, respectively. In 79 high-grade squamous intraepithelial lesions (CINⅡ and Ⅲ) and cervical cancer tissues, 50 of 79 were infected with HPV16/18, while 29 of 79 were negative. The methylation ratio ofDBC1 gene was 88.0% in HPV16/18 infection positive group while the methylation ratio was 55.2% in negative group (P<0.05). The expression ofDBC1 gene at mRNA level in 10 methy- lation-positive cervical cancer tissues was significantly lower than that in the 10 methylation-negative normal cervical tissues (P<0.05).Conclusion:The methylation ofDBC1 gene may become a molecular marker to detect cervical cancer of Uyghur women in Xinjiang.DBC1 gene methylation combined with HPV16/18 infection test can be used to aid diagnosis of cervical cancer.

Objective:To describe the trend of mortality and death spectrum in Shandong Province from 1970 to 2021 and provide basis for the targeted disease prevention and control.Methods:The data were collected from the death registration reports of Shandong and 3 national retrospective surveys of death causes in Shandong. The change in levels of overall and specific deaths in Shandong in different years were analyzed based on mortality rate, age-standardized mortality rate and constituent ratio of cause of death, differential decomposition was used to quantify the contribution of demographic and non-demographic factors to changes of mortality.Results:The crude mortality rate in residents in Shandong was basically stable from 1970 to 2021, and the mortality rate during 2020-2021 (732.73/100 000) was slightly higher than that during 1970-1974 (671.98/100 000). While the standardized mortality rate decreased significantly, and the mortality during 2020-2021 (183.39/100 000) decreased by 67.71% compared with that during 1970-1974 (568.00/100 000). The negative increase of population factors and the positive decrease of non-population factors reacted each other, so the mortality was relatively stable. Cardiac-cerebrovascular disease was always the leading cause of death, but the constituent ratio of death increased rapidly from 19.70% during 1970-1974 to 54.72% during 2020-2021. The rank in the causes of death changed from the fourth (11.46%) to the second (25.70%) for malignant tumor, from the seventh (5.85%) to the third (5.59%) for injury, from the second (12.87%) to the fourth (4.99%) for chronic respiratory diseases, from the third (12.27%) to the tenth (0.42%) for infectious diseases. The standardized mortality rates of the main causes of death decreased at different degrees, the standardized mortality rates of obstetrical disease, infectious disease, gastrointestinal disease and chronic respiratory disease decreased by more than 50.00%. The age distribution of deaths and the death spectrum in different age groups and in urban-rural populations changed significantly. During 2020-2021, the proportion of deaths in young people aged 0-14 years was 0.54%, which was 97.05% lower than that during 1970-1974, while the proportion of deaths in the elderly aged ≥75 years was 55.14%, which was 55.75% higher than that during 1970-1974. The rank of infectious diseases in the causes of death descended significantly in all age groups, but the ranks of injury, neuropsychiatric disease and malignant tumor rose significantly in adolescents, and the ranks of endocrine nutrition and metabolic disease rose in middle-aged and elderly people. The difference of death spectrum between urban area and rural area became less obvious and the main death causes in urban and rural residents were basically the same during 2020-2021.Conclusions:The death spectrum of residents in Shandong changed significantly. Chronic and non-communicable diseases, especially cardiac-cerebrovascular disease and malignant tumor, should be the focus in disease control and prevention. The prevention and control of diseases in Shandong made remarkable achievement during 1970-2021. However, in the context of population ageing, it is suggested to strengthen the treatment, prevention of diseases and injuries related to the health of the elderly and elderly health care in the future.

Objective:To understand the characteristics and trends of acute myocardial infarction (AMI) in Shandong Province and to provide evidence for formulating prevention and control strategies.Methods:Data were derived from the AMI incidence reports of Shandong Province's Chronic Disease Surveillance Information Management System in 2012-2021. The crude and standardized incidence rates were used as indicators to describe the incidence level of AMI. Joinpoint regression analysis was used to analyze the trends in the incidence and age of onset over the years. The contribution of population aging to the increase in AMI incidence was assessed using the rate difference decomposition method. The incidence of AMI in each district (county) in Shandong Province was visualized using ArcGIS 10.8 software, and global and local spatial autocorrelation analysis was performed using DeoDa 1.12 software.Results:From 2012 to 2021, 198 233 cases of AMI were reported from 19 provincial monitoring sites in Shandong Province, of which 53.13% were males and 97.12% were ≥45 years old. The reported crude incidence increased from 90.12 per 100 000 in 2012 to 176.54 per 100 000 in 2021, with an average annual increase of 7.01% ( Z=7.35 , P<0.001). There was no significant upward trend in standardized incidence ( Z=1.64 , P=0.140), but the standardized incidence of male residents showed an increasing trend ( Z=2.76 , P=0.028). Before 2014, the reported crude incidence of males was similar to that of females, but after 2014, the reported crude incidence of males was continuously higher than that of females. However, males' standardized incidence was higher than females in all years. Both crude and standardized incidence rates were higher in rural residents than in urban areas. The median onset of AMI increased from 71.6 years old in 2012 to 73.5 years old in 2021. The median age of onset in males was lower than that in females in all years, and in most years, the median age of onset in urban residents was lower than that in rural residents. The incidence of AMI in males showed a trend in younger age groups. According to the seasonal decomposition, the incidence peak of AMI was in January, and the trough was in September. The contribution of aging population to the increase in crude incidence of AMI increased from 8.63% in 2013 to 52.58% in 2021. The global spatial autocorrelation analysis showed that the incidence of AMI presented an obvious spatial clustering distribution. Local spatial autocorrelation analysis found that the high-incidence areas (counties) were mainly concentrated in Liaocheng City and Dezhou City in the northwest region of Shandong Province and Heze City in the southwest. Conclusions:The incidence of AMI among residents in Shandong Province was rising, with spatial clustering and seasonal clustering characteristics. People aged 45 years and older, male residents, and rural residents were at high risk of developing AMI. There was a certain trend of younger age at onset among men. Targeted prevention and control measures should be taken for high-incidence seasons, high-risk groups, and high-incidence clustering areas in northwestern Shandong Province.

Objective:To optimize the stimulation and activation system of mouse CD3 + T cells in vitro and explore the optimal infection time of CD3 + T cells to establish mouse CD19 chimeric antigen receptor T cells (mCD19 CAR-T) , and to also verify its killing effect in vivo and in vitro. Method:Splenic CD3 +T cells were isolated and purified using magnetic beads, and the cells were cultured in Soluble anti-CD3/CD28, PMA+Ionomycin, and Plated anti-CD3/CD28. Cell activation and apoptosis were assessed by flow cytometry after 8, 24, 48, and 72 hours. ScFv plasmid of mouse CD19 antibody was transfected to plat-E cells to package retrovirus. Activated CD3 + T cells were infected to construct mouse-specific CD19 chimeric antigen receptor T cells (mCD19 CAR-T) , and mCD19 CAR-T cells were co-cultured with B-cell lymphoma cell line A20 in vitro. The specific toxicity of A20 was detected by flow cytometry, and mCD19 CAR-T cells were infused into the lymphoma mouse model to detect its killing effect and distribution. Results:The activation effect of Plated anti-CD3/CD28 on CD3 + T cells was superior, with the cells exhibiting good viability 24–48 hours after stimulation. Established mCD19 CAR-T cells with stable efficiency[ (32.27±7.56) % ] were specifically able to kill A20 tumor cells (The apoptosis rate was 24.3% at 48 h) . In vivo detection showed a non-significant decrease in the percentage[ (1.83±0.58) % ] of splenic CD19 + cells 6 days after mCD19 CAR-T cell infusion. A marked clearance in bone marrow and spleen appeared on day 12 compared with the A20 group, and this difference was statistically significant[spleen: (0.36±0.04) % vs (47.00±13.46) % , P<0.001; bone marrow: (1.82±0.29) % vs (37.30±1.44) % , P<0.0001]. Moreover, mCD19 CAR-T cells were distributed in high proportions in the peripheral blood, spleen, and bone marrow[ (2.90±1.12) % , (4.96±0.80) % , (13.55±1.56) % ]. Conclusion:This study demonstrated an optimized activation system and the optimal infection time of CD3 + T cells. Furthermore, stable constructed mCD19 CAR-T cells showed a remarkable killing ability in vitro and in vivo.

Objective To investigate the pattern of lymph node metastasis and analyze prognostic factors of Siewert type Ⅱ adenocarcinoma of esophagogastric junction (AEG).Methods The retrospective case-control study was conducted.The clinicopathological data of 368 patients with Siewert type Ⅱ AEG who were admitted to Tianjin Medical University Cancer Institute and Hospital from June 2010 and November 2015 were collected.There were 323 males and 45 females,aged from 35 to 80 years,with an average age of 64 years.Of 368 patients,209 underwent left transthoracic surgery,1 12 underwent thoracoabdominal surgery,and 47 underwent Ivor-Lewis surgery.Observation indicators:(1) total lymph node metastasis and metastasis of various lymph node stations;(2) follow-up and survival;(3) prognostic factors analysis;(4) influencing factors affecting thoracic lymph node metastasis.Follow-up using outpatient examination and telephone interview was performed to detect survival of patients up to November 2018.Measurement data with skewed distribution were represented as M (range).Count data were represented as absolute number or percentage.The survival time and rate were calculated using the Kaplan-Meier method.The univariate and multivariate analyses were done by the COX proportional hazard model.Results (1) Total lymph node metastasis and metastasis of various lymph node stations:the total lymph node metastasis rate was 66.58％ (245/368) in 368 patients.The metastasis rates of abdominal lymph nodes,thoracic lymph nodes,lower mediastinal lymph nodes,and upper mediastinal lymph nodes were 65.49％ (241/368),12.77％ (47/368),12.23％ (45/368),and 1.09％ (4/368),respectively.The order of metastasis rate of various lymph node stations from high to low was 51.99％(170/237) of No.7 left gastric artery,34.23％(89/260) of No.1 right paracardial region,33.88％ (83/245) of No.2 left paracardial region,28.91％ (85/294) of No.3 lesser curvature,27.10％(29/107) of No.1 1 splenic artery,19.75％(16/81) of No.9 celiac trunk,15.25％(36/236) of No.E8Lo lower paraesophageal region,11.94％ (16/134) of No.4 greater curvature,11.76％ (6/51) of No.E8M middle paraesophageal region,11.1 1％(10/90) of No.8 common hepatic artery,4.65％(4/86) of No.E9L left inferior pulmonary ligament and 3.39％ (2/59) of No.E7 subcarinal region.(2) Follow-up and survival:of the 368 patients,309 were followed up for 1-103 months,with a median follow-up time of 38 months.The survival time of 309 patients was 0.7-101.9 months,and the median survival time was 35.9 months.During the followup,the postoperative l-,2-,3-year overall survival rates were 85.9％,68.6％,and 58.7％,respectively.(3) Prognostic factors analysis.Results of univariate analysis showed that tumor differentiation degree,presence of thoracic lymph node metastasis,number of metastatic lymph nodes,T staging,tumor diameter,and length of esophageal invasion were associated factors affecting prognosis of patients (x2 =8.776,26.582,46.057,18.679,22.460,9.158,P＜0.05).Results of multivariate analysis showed that presence of thoracic lymph node metastasis,number of metastatic lymph nodes,T staging,and tumor diameter were independent influencing factors for prognosis of patients [odds ratio (OR) =1.699,1.271,1.422,1.238,95％ confidence interval:1.102-2.621,1.019-1.481,1.090-1.856,0.971-1.481,P＜0.05].(4) Influencing factors affecting thoracic lymph node metastasis:results of univariate analysis showed that tumor diameter,length of esophageal invasion,number of lymph lodes harvested in thorax were related factors for thoracic lymph node metastasis (x2 =5.129,43.140,10.605,P＜0.05).Results of multivariate analysis showed that length of esophageal invasion ≥2 cm,number of lymph lodes harvested in thorax ≥ 4 were independent risk factors for thoracic lymph node metastasis (OR =6.321,1.097,95％ confidence interval:2.982-13.398,1.026-1.173,P＜0.05).Conclusion Lymph node metastasis of Siewert type Ⅱ AEG spreads two regions,mainly at abdominal lymph nodes,followed by the thoracic lymph nodes.Presence of thoracic lymph node metastasis,number of metastatic lymph nodes,T staging,and tumor diameter are independent influencing factors for prognosis of patients.Presence of thoracic lymph node metastasis indicates poor prognosis of patients.Length of esophageal invasion ≥ 2 cm and number of lymph lodes harvested in thorax ≥4 are independent risk factors for thoracic lymph node metastasis.

OBJECTIVE To establish the quality standard of Triadica cochinchinensis and to perform the acute toxicity study. METHODS Appearance properties, powder microscopic identification, and thin-layer chromatography(TLC) identification were researched. The specific chromatogram was established by HPLC. The content of cadmium(Cd), lead(Pb), arsenic(As), copper(Cu), and mercury(Hg) was determined by inductively coupled plasma-mass spectrometry(ICP-MS). Acute toxicity was studied by maximum dose. RESULTS The outer skin of herbs was dark brown, and the inner surface was light yellow brown and fibrous. Besides, crystal sheath fiber was common, and calcium oxalate clusters arranges in rows. In the TLC diagram of the test product, the fluorescent spots of the same color were displayed at the corresponding position of the control product(scopoletin, isofraxidin). Five common peaks were calibrated in the characteristic map and the three characteristic peaks(scopoletin, isofraxidin, dimethylfraxetin) were recognized. The content of the measured heavy metal elements was lower than the national limit standard. The linear correlation coefficient was R2 > 0.999. The precision, stability, repetitive RSD were < 10%. The average recovery rate of the added sample was 80%−120%, and the RSD was < 10%. The maximum dose of the acute toxicity test was 184.09 g·kg−1. The 14 d internal body mass, food intake, organ-body ratios, the serum glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, blood urea nitrogen, and creatinine were not significantly different by comparing with the normal controls. Therefore, no significant toxicity was observed. CONCLUSION The established standard can provide a reference for evaluating the quality of Triadica cochinchinensis. The heavy metal content of ten batches of medicinal materials is within the safe range. Acute toxicity test show that there is no obvious significant adverse teactions after oral administration, and the safe dose range is large, which can provide a reference for the subsequent development and utilization.