The aim of this study is to establish stable transfected cell lines which could produce SPAG4L protein labeled with Myc and His tags in vitro. The open reading frame (ORF) of human SPAG4L was amplified by PCR and the fragments were cloned into eukaryotic expression vector pcDNA3.1/myc-His(-)A. The recombined plasmids of pcDNA3.1/myc-His(-)A/SPAG4L were verified by sequencing and digestion with enzymes. Then, the recombined plasmids were introduced into HeLa cells and screened by G418. Western blotting was performed to detect the expression of SPAG4L and its tags in stable transfected cell lines. SPAG4L and its tags were expressed in the stable cell lines transfected with pcDNA3.1/myc-His(-)A/SPAG4L, but not in the control group. Further study showed that SPAG4L colocalized with the endoplasmic reticulum(ER) marker PDI by immunofluorescence. The stable transfected cell lines established in this study will provide a powerful tool for further studies such as co-immunoprecipitation and pull-down.
Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Female ; Genetic Vectors ; genetics ; HeLa Cells ; Histidine ; genetics ; Humans ; Male ; Proto-Oncogene Proteins c-myc ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Transfection

Objective: To learn about the construction and staffing of the school health system in Chinese institutions for disease prevention and control, and to provide basic information for the school health system, team capacity building and work development. Methods: Electronic questionnaire was used to collect the setting and staffing of school health departments (including school health centers and departments/rooms) at the provincial, prefecture and county (district) levels in the centers for disease control and prevention. Statistical analysis was made on the proportion of school health, the number of staff and the characteristics such as age, education, major and working years in the provincial, prefecture and county (district) levels. Results: Among the 3 313 institutions, the proportion of independent school health departments was 10.8%, and those of the provincial, prefecture and county (district) levels were 74.2%, 15.0%, and 9.6%, respectively. Among the institutions with separated department, the average number of staff members was 4.4, while the number of staff was 2.5. The average age of school health workers was 40.4 years old, and the proportion of male and female employees was 45.2% and 54.8%. The proportion of personnel who have been engaged in school health work for less than 5 years on average was as high as 65.1%. The majors of the staff were mainly public health ( 40.4 %), 54.0% of the provincial staff had a master s degree or above, and 47.8% and 58.7% of the staff at the prefecture and county (district) levels were junior college or below respectively.The proportion of provincial level personnel with intermediate and senior titles was 69.6%, and the proportion of municipal and countylevel personnel at the junior level and below was 52.2% and 56.2% respectively. Conclusion The proportion of independent school health departments within centers of disease control and prevention across China was low. There is a serious shortage of school health personnel, and there are problems such as low levels of education and professional titles, especially in county (district) level institutions. It is urgent to strengthen the construction of the school health system of the centers for disease control and prevention in China.

Kidney stones are mixed by various inorganic salts and organic matter according to certain rules. The process of crystal nucleation, growth and aggregation is the key step of kidney stone formation. The different crystal structures will bring about the different formation process and physicochemical properties of kidney stones. It is of great significance to study the crystal structures and formation characteristics of kidney stone to clarify the causes of it and prevent the recurrence of it. In this paper, based on the microstructure and crystal structure of kidney stones, the distribution of different crystals and components in kidney stones, the nucleation and growth process of crystal forming kidney stones, and the different treatment methods based on crystal structure are reviewed in recent years.

Objective:To investigate the association between cancer fatalism and optimistic attitudes among colorectal cancer patients, and how the above linkage is moderated by the involvement of patients′ opinions, the family members′ opinions and the physicians′ opinions in treatment decision-making.Methods:A cross-sectional survey was conducted on 64 patients diagnosed with colorectal cancer and awaiting radical colorectal cancer surgery in the Department of General Surgery of Beijing Friendship Hospital Affiliated to Capital Medical University, from January 2021 to December 2021. There were 38 males and 26 females, aged (61.2±13.0) years from 30 to 84 years. Cancer fatalism, optimism attitudes, and the involvement of patients′ opinions, the family members′ opinions and the physicians′ opinions in treatment decision-making were assessed. The patients′ cancer fatalism beliefs were measured by the " Chance" subscale of the Form C of the Multidimensional Health Locus of Control (MHLC-C), optimism attitudes were assessed by the Chinese version of the revised Life Orientation Test (CLOT-R), and the influence of patients′, family members′, and physicians′ opinions in the medical decision-making process was measured by a self-designed single factor scale. Measurement data of normal distribution were expressed as mean±standard deviation ( ± s). Measurement data of skewed distribution were expressed as M( Q1, Q3). Spearman correlation analysis and Chi-square test was used to examine the association between the main outcome variable (patients′ optimistic attitudes) and demographic and clinical characteristics. Interaction was examined by hierarchical linear regression analysis combined with simple slope tests. Results:Cancer fatalism was negatively associated with patients′ optimistic attitudes ( r=-0.35, P<0.01). Optimistic attitude of patients was significantly and negatively correlated with the influence of family members′ opinions on medical decision-making ( r=-0.25, P<0.05). There were significant positive correlations between the influence of patients′ and family members′ opinions ( r=0.50, P<0.01), family members′ and physicians′ opinions ( r=0.67, P<0.01), and physicians′ and patients′ opinions ( r=0.38, P<0.01) in medical decision making. Hierarchical linear regression analysis showed a negative association between cancer fatalism and optimism ( β=-0.32, P=0.01). This association was further moderated by the involvement of family members′ opinions ( β=-0.56, P<0.01) and the involvement of physicians′ opinions ( β=-0.36, P=0.04) in medical decision-making. Simple slope tests revealed that the negative impact of fatalistic attitudes on patients′ optimism attitudes may be potentiated when family members′ opinions have high influence on medical decision-making, while the negative impact may be buffered to some extent when physicians′ opinions have high influence on medical decision-making. Conclusions:Cancer fatalism had a negative effect on patients′ optimism. The high influence of physicians in treatment decision-making buffered the negative effect of cancer fatalism on optimism; the high influence of family members in treatment decision-making potentiated the negative effect of cancer fatalism on optimism. In the process of doctor-patient communication and shared decision-making, for patients with strong fatalistic attitudes, consideration should be given to appropriately increasing direct informational communication between physicians and patients and reducing excessive family intervention in medical decision-making, so as to enhance patients′ autonomy for treatment, promote optimism, and reduce the negative effects of cancer fatalism.

Objective: To understand the current status of main professional work in independent school health departments of Chinese centers for disease control and prevention, so as to provide reference and suggestions for the further development of school health work in China. Methods: Electronic questionnaire was used to collect the basic work of school health, the monitoring work, the intervention action of common diseases and the development of health intervention among students in independent school health departments of centers for disease control and prevention in China. Results: Among the 357 institutions that have set up independent school health departments, the implementation rates of school mental health work, safety emergency and risk avoidance health intervention were low, which were 11.8% and 11.5%, respectively. Relying on the project "national monitoring and intervention of common diseases and health influencing factors of students", the overall implementation of health monitoring in schools nationwide was successful, but the overall implementation rate of students nutritional status monitoring and "healthy parents action" were low, accounting for 44.5% and 24.4%, respectively. At the same time, there were still as many as 27.2% institutions that had not carried out the intervention action for common diseases of students which advocated in the monitoring program. The failure rate of county level institutions was higher than that of provincial level and prefecture level institutions, and the failure rate of the central and western institutions was much higher than that of the eastern institutions; the difference was statistically significant( χ 2=30.1, 41.6, P <0.05). Conclusion We should increase support including policy preference, fund guarantee, technical guidance and so on for the school health work of disease control institutions at the grass roots level and in economically underdeveloped areas, so as to ensure the healthy growth of children and adolescents in all respects.

Objective:To explore whether there are gender differences in clinical and pathological characteristics and prognosis of young patients with rectal cancer (under 50 years old), and to analyze the risk factors affecting the prognosis of young patients with rectal cancer.Methods:The medical records of 85 young rectal cancer patients admitted to Beijing Friendship Hospital Affiliated to Capital Medical University from January 2015 to December 2020 were retrospectively collected. According to gender, they were divided into male group ( n=50) and female group ( n=35). The age was (43.67±5.50) years old, ranging from 26 to 50 years old. Primary outcome measures were sex, disease-free survival, and overall survival. Secondary outcomes were family history, body mass index (BMI), clinical stage, anemia, whether the female patient was menopausal, whether the female patient took oral estrogen, the location of the primary lesion, whether neoadjuvant therapy was performed, pathological stage, whether accompanied with vascular nerve invasion, and whether postoperative adjuvant therapy was performed. R4.0.2 software was used for statistical analysis. The measurement data with normal distribution in the collected data were expressed as mean±standard deviation ( ± s), and the comparison between groups was analyzed by t test. Count data were expressed as constituent ratio, and analyzed using the chi-square test or Fisher′s exact test. The survival curve was drawn by Kaplan-Meier method, and the difference in survival rate was tested by Log-rank test. Factors with statistical significance in univariate analysis were included in COX proportional regression model for multivariate analysis to screen independent risk factors affecting overall survival. Results:Compared with male patients, a higher proportion of young female patients with rectal cancer were diagnosed with anemia before surgery (42.9% vs 22.0%, P=0.040). The 1-year, 3-year and 5-year overall survival rates were 94.3%, 80.0% and 68.6% in young female patients, and 98.0%, 90.0% and 90.0% in young male patients, respectively. The median disease-free surival was 31.6 months for women and 34.4 months for men. Multivariate analysis showed that female( HR=3.799, 95% CI: 1.312-11.002, P=0.014)and BMI( HR=0.846, 95% CI: 0.724-0.989, P=0.036)were independent risk factors affecting the prognosis of young patients with rectal cancer. Conclusions:Young female patients have a worse prognosis than male patients. Female and BMI are independent risk factors for the prognosis of young rectal cancer patients, and gender should be the key research object of observation in young rectal cancer patients.

Objective To investigate the efficacy of drug-coated balloon in treatment of femoropopliteal artery occlusive disease.Methods The clinical data of 49 patients with femoropopliteal artery occlusive disease from Feb 2016 to Oct 2017 were retrospectively analyzed.25 cases underwent drugcoated balloon treatment (DCB group),and 24 cases received plain balloon angioplasty (POBA group).Results The procedures were successful in all cases.There were no amputation or death occurred.In DCB group compared to POBA group,the patency rate at postoperative 3 months were not significantly different (96.0％ vs.87.5％,P ＞ 0.05),while it was significantly different at postoperative 6 and 12 months (88.0％ vs.66.7％;80.0％ vs.50.0％,both P ＜0.05).Minimal lumen diameter (MLD) changes and late lumen loss(LLL):The MLD in preoperative,immediate postoperative was not statistically significant between the three group(P ＞ 0.05).At 6 months and 12 months after operation,MLD of DCB group was higher than POBA group (P ＜ 0.05).At 12 months after surgery,LLL in DCB group was significantly lower than the POBA group (P ＜ 0.05).Conclusion DCB is the effective method to treat femoropopliteal artery occlusive disease,which has better short and mid-term efficacy than POBA.

Oxalate is an organic dicarboxylic acid that is a common component of plant foods.The kidneys are essential organs for oxalate excretion,but excessive oxalates may induce kidney stones.Jupiter micro-tubule associated homolog 2(JPT2)is a critical molecule in Ca2+mobilization,and its intrinsic mecha-nism in oxalate exposure and kidney stones remains unclear.This study aimed to reveal the mechanism of JPT2 in oxalate exposure and kidney stones.Genetic approaches were used to control JPT2 expression in cells and mice,and theJPT2 mechanism of action was analyzed using transcriptomics and untargeted metabolomics.The results showed that oxalate exposure triggered the upregulation of JPT2,which is involved in nicotinic acid adenine dinucleotide phosphate(NAADP)-mediated Ca2+mobilization.Tran-scriptomic analysis revealed that cell adhesion and macrophage inflammatory polarization were inhibited by JPT2 knockdown,and these were dominated by phosphatidylinositol 3-kinase(PI3K)/AKT signaling,respectively.Untargeted metabolomics indicated that JPT2 knockdown inhibited the produc-tion of succinic acid semialdehyde(SSA)in macrophages.Furthermore,JPT2 deficiency in mice inhibited kidney stones mineralization.In conclusion,this study demonstrates that oxalate exposure facilitates kidney stones by promoting crystal-cell adhesion,and modulating macrophage metabolism and in-flammatory polarization via JPT2/PI3K/AKT signaling.

Objective:To explore the relationship between the occurrence and development of colorectal cancer and the risk of BRCA1 and BRCA2 gene mutations. Methods:Sixty-one patients with colorectal adenocarcinoma admitted to Beijing Friendship Hospital Affiliated to Capital Medical University from January 2022 to March 2022 were tested by second-generation sequencing. Information such as age of onset, gender, histological grade and TNM stage were collected. According to whether the BRCA1 and BRCA2 gene had systemic mutation, the patients were divided into BRCA1 and BRCA2 gene system mutation group and unmutated group. There were 8 cases in the BRCA1 and BRCA2 gene system mutant group and 53 cases in the non-mutant group. The main outcome measures were the relationship between pathogenic or likely pathogenic germline mutations in colorectal cancer and clinicopathological data of patients, including age of onset, gender, tumor location, TNM stage, histological differentiation, and family history. The secondary outcome was the relationship between BRCA1 and BRCA2 gene system mutations and clinicopathological data. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and comparison between groups was analyzed using the t-test. Measurement data with non-normal distribution were represented as M ( Q1, Q3), and comparison between groups was analyzed using the Mann-Whitney U test. Measurement data were expressed as the number of cases or percentage (%), and Chi-square test was used for comparison between groups. Results:Among 61 colorectal adenocarcinoma patients, the frequency of pathogenic or potentially pathogenic germ line mutations in colorectal cancer was 13.1% (8/61), and the frequency of BRCA1 and BRCA2 mutations was 3.3% (2/61). The frequency of BRCA1 and BRCA2 mutations was 13.1% (8/61). Women with BRCA1 and BRCA2 mutations (75.0% vs 37.7%, χ2=3.947, P=0.047) and right colon cancer (75.0% vs 26.4%, χ2=7.889, P=0.019) were significantly higher than those without mutation. Conclusions:The frequency of BRCA1 and BRCA2 gene mutation is higher in colorectal cancer patients. BRCA1 and BRCA2 gene mutations are recommended for colorectal cancer patients with a family history of breast or ovarian cancer.

Objective:To explore the risk factors associated with liver metastasis in T 1 stage colorectal neoplasms patients and establish a prognostic model. Methods:Clinicopathological data of T 1 stage colorectal neoplasms patients from the Surveillance, Epidemiology, and End Results (SEER) database between January 1, 2004, and December 31, 2019 were included. The differences in clinicopathological characteristics between patients with and without liver metastasis were compared using Chi-square test, Wilcoxon rank-sum test, and multivariate Logistic regression analysis. Survival curves were plotted using Kaplan-Meier method, and the Log-Rank test was used to compare survival differences between the two groups. Univariate and multivariate analysis of prognostic factors were performed using the proportional Cox regression hazards regression model. Patients were randomly divided into training set and validation set at a ratio of 6∶4 using simple random sampling method. A nomogram model was established based on independent prognostic factors based on the results of the multivariate Cox regression analysis. The predictive ability of the model was evaluated using time-dependent receiver operating characteristic (ROC) curves and calibration curves. Results:A total of 28 258 T 1 stage colorectal neoplasms patients were included in the study. The multivariate Logistic regression analysis for liver metastasis occurrence indicated that the neuroendocrine neoplasms, elevated carcinoembryonic antigen (CEA) levels, larger neoplasms size, positive lymph node metastasis, and presence of cancer nodules were statistically significant differences ( P<0.05). For T 1 stage colorectal neoplasms patients with liver metastasis, the results of the multivariate Cox regression analysis showed that age, primary site, ethnicity, chemotherapy, and surgical treatment were independent prognostic factors ( P<0.05). The nomogram constructed based on these five prognostic factors had time-dependent ROC areas under the curve of 0.758, 0.797 and 0.729 for 1-year, 3-year, and 5-year survival rates, respectively, 0.749, 0.857, 0.871 in the validation set. The calibration curves of the nomogram in the training and validation sets were close to the 45-degree diagonal line. Conclusions:Neuroendocrine neoplasms, elevated CEA levels, larger neoplasms size, positive lymph node metastasis, and presence of cancer nodules are independent risk factors for liver metastasis in T 1 stage colorectal neoplasms. Age, primary site, ethnicity, chemotherapy, and surgical treatment are independent prognostic factors. The nomogram constructed based on these clinical characteristics has good discrimination and calibration abilities.