Objective:To investigate the role of the Smad3 signaling pathway in the process of silica-induced epithelial-mesenchymal transition (EMT) .Methods:In September 2022, lung epithelial cells (BEAS-2B) were exposed to different concentrations of silica suspension (0, 50, 100, and 150 μg/ml) for 6 and 12 hours. Additionally, SIS3, a specific inhibitor of phosphorylated Smad3 (p-Smad3) , was utilized to establish the p-Smad3 inhibition model. The cells were divided into four groups: blank control gruop, silica group, SIS3 intervention group, and SIS3 +silica group. Cell morphology was observed using an inverted fluorescence microscope, while cell viability was assessed using a Cell Counting Kit-8 (CCK-8) . The mRNA and protein expression levels of E-cadherin (E-Cad) , N-cadherin (N-Cad) , Vimentin, Smad3, and p-Smad3 were analyzed by Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Differences between two groups were compared using Student's t-test, and multiple group comparisons were analyzed using a one-way analysis of variance with the Student-Newman-Keuls test.Results:Compared with the blank control group, the morphology of BEAS-2B cells shifted from epithelial to mesenchymal cell-like following silica exposure, and the cell viability of BEAS-2B cells declined after exposure to 150 μg/ml silica for 6 and 12 hours. Furthermore, silica exposure led to significant reductions in mRNA and protein expression levels of the epithelial cellular marker (E-Cad) in BEAS-2B cells, accompanied by increased expressions of interstitial cellular markers (N-Cad and Vimentin) . Importantly, the level of p-Smad3/Smad3 expression levels was also elevated in silica-treated cells ( P<0.05) . Compared to the blank control group, the level of p-Smad3/Smad3 expression levels was significantty reduced. Moreover, compared to the silica group, the protein expression levels of N-Cad and Vimentin in the cell of the SIS3+silica group were significantly reduced, while the E-Cad expression was increased ( P<0.05) . Conclusion:Silica exposure can prmote the epithelial mesenchymaol transformotion process by activating smod3 signa ling pathuay, and in hibiting smad3 signa ling pathuay can effctively alleviate the occurrence of epithelial mesenchymal transformation process.

Objective:To investigate the risk factors of secondary primary cancer (SPC) after cervical cancer and to construct a nomogram model for predicting the prognosis.Methods:The data of 3 790 patients with primary cervical cancer from the 3rd edition of the International Classification of Diseases Oncology (ICD-O-3) number C53.9 between 2000 and 2020 in the Surveillance, Epidemiology and End Results (SEER) database were retrospectively analyzed, and SPC occurred in 2 036 cases out of 3 790 patients. Standardized incidence rate (SIR) of patients with cervical cancer was calculated by using SEER*Stat software; Cox proportional hazards model was used to make the univariate and multivariate analysis on the influencing factors of the overall survival (OS) in patients with SPC after cervical cancer. By using the "rms" package of R software, nomogram models for predicting 1-, 3-, and 5-year OS rates in SPC patients after cervical cancer were constructed based on prognostic independent influencing factors. The prediction efficacy and consistency of the model were verified by using the receiver operating characteristic (ROC) curve and the calibration curve.Results:The cervical cancer patients with the age of diagnosis of 20-34 years old (SIR: 1.76, 95% CI: 1.57-1.98), black race (SIR: 1.75, 95% CI: 1.61-1.90), the interval period of 2 primary tumors: 6-11 months (SIR: 1.69, 95% CI: 1.50-1.90), histologic grade Ⅳ (SIR: 1.62, 95% CI: 1 30-2.00), chemotherapy (SIR: 1.63, 95% CI: 1.56-1.71), radiotherapy (SIR: 1.59, 95% CI: 1.53-1.66), unmarried (SIR: 1.47, 95% CI: 1.41-1.54) had high SIR. Multivariate Cox regression analysis results of 2 036 SPC patients after cervical cancer showed that unmarried, SEER stage of regional lesion phase, distant metastasis phase and unknown, histologic grade of Ⅱ, Ⅲ, Ⅳ and the unknown, unknown lymph node dissection, other sites expert for lymphoma in SPC sites were independent risk factors of OS in SPC patients after cervical cancer (all P < 0.05); receiving surgery for SPC (compared to those not receiving surgery, HR = 0.38, 95% CI: 0.32-0.45, P = 0.001), radiotherapy (compared to those not receiving radiotherapy or unknown, HR = 0.66, 95% CI: 0.56-0.78, P = 0.001), and chemotherapy (compared to those not receiving chemotherapy or unknown, HR = 0.86, 95% CI:0.74-0.99, P = 0.034) were independent protective factors of OS in SPC patients after cervical cancer. The results of Kaplan-Meier survival analysis showed that the differences in the OS of SPC patients after cervical cancer with different marriage status, SEER stage, histologic grade, lymph node dissection, surgery for SPC, primary sites of SPC and whether receiving radiochemotherapy were all statistically significant (all P < 0.001). Based on the 8 variables including marital status, SEER stage, histologic grade, whether lymph nodes have been cleared, whether SPC has been treated with surgery, radiotherapy, chemotherapy, and the primary sites of SPC, a nomogram model for predicting the 1-, 3- and 5-year OS rates of SPC patients after cervical cancer was established. The results of ROC curve analysis showed that the area under the curve of the model for predicting 1-, 3-, and 5-year OS rates was 0.841, 0.847, and 0.847, respectively. The calibration curves showed a good consistency between the predicted results of model and the actual results. Conclusions:A prognostic prediction nomogram model for SPC after cervical cancer constructed based on the data in the SEER database has a high clinical application value and calibration.

Objective To investigate the relationship between stationing years of personnel on an island and their traditional Chinese medicine(TCM)constitution,thus providing a reference for adjusting the health status of stationed personnel,preventing and treating diseases.Methods Based on The Scale of Constitution in Chinese Medicine Questionnaire,TCM constitution of 734 personnel stationed on an island was investigated.Pearson χ2 method was used for data analysis.Results Of the 734 personnel stationed on an island 345(47.0%)were of the balanced constitution type and 389(53.0%)were of the biased constitution types.Among the people with biased constitution types,composite constitution accounted for 80.5%(313 people)and the simple accounted for the rest(19.5%,76 people).The top three types of the biased were dampness heat(15.2%),qi deficiency(14.0%),and yin deficiency(10.7%).The distribution of TCM constitution types was significantly different in terms of stationing years on the island(P<0.05).There was a significant difference in the balanced type and yin deficiency type between people with stationing time≤2 years and>8 years(P<0.05).There was a significant difference in qi deficiency type and qi depression type between people with stationing time≤2 years and>5 years(P<0.05).Compared to people with stationing time≤2 years,significant difference was found in yang deficiency type in people with stationing time ranging from 2 to 5 years and those with>8 years(P<0.05).There were significant differences in the phlegm dampness type,blood stasis type,and specific diathesis type between people with stationing time≤2 years and people with stationing time ranging from>2～5 years and>8 years(P<0.05).There were significant differences in the dampness heat type between people with stationing time≤2 years and people with stationing time ranging from>5～8 years and>8 years,between pepole with stationing time ranging from>2～5 years and people with stationing time>8 years(P<0.05).Conclusion The dampness heat type,qi deficiency type and yin deficiency type are common biased TCM constitution in personnel stationed on islands.The longer the time spent on islands,the greater the possibility of forming biased constitution.

Objective:To investigate the risk factors of secondary primary cancer (SPC) after cervical cancer and to construct a nomogram model for predicting the prognosis.Methods:The data of 3 790 patients with primary cervical cancer from the 3rd edition of the International Classification of Diseases Oncology (ICD-O-3) number C53.9 between 2000 and 2020 in the Surveillance, Epidemiology and End Results (SEER) database were retrospectively analyzed, and SPC occurred in 2 036 cases out of 3 790 patients. Standardized incidence rate (SIR) of patients with cervical cancer was calculated by using SEER*Stat software; Cox proportional hazards model was used to make the univariate and multivariate analysis on the influencing factors of the overall survival (OS) in patients with SPC after cervical cancer. By using the "rms" package of R software, nomogram models for predicting 1-, 3-, and 5-year OS rates in SPC patients after cervical cancer were constructed based on prognostic independent influencing factors. The prediction efficacy and consistency of the model were verified by using the receiver operating characteristic (ROC) curve and the calibration curve.Results:The cervical cancer patients with the age of diagnosis of 20-34 years old (SIR: 1.76, 95% CI: 1.57-1.98), black race (SIR: 1.75, 95% CI: 1.61-1.90), the interval period of 2 primary tumors: 6-11 months (SIR: 1.69, 95% CI: 1.50-1.90), histologic grade Ⅳ (SIR: 1.62, 95% CI: 1 30-2.00), chemotherapy (SIR: 1.63, 95% CI: 1.56-1.71), radiotherapy (SIR: 1.59, 95% CI: 1.53-1.66), unmarried (SIR: 1.47, 95% CI: 1.41-1.54) had high SIR. Multivariate Cox regression analysis results of 2 036 SPC patients after cervical cancer showed that unmarried, SEER stage of regional lesion phase, distant metastasis phase and unknown, histologic grade of Ⅱ, Ⅲ, Ⅳ and the unknown, unknown lymph node dissection, other sites expert for lymphoma in SPC sites were independent risk factors of OS in SPC patients after cervical cancer (all P < 0.05); receiving surgery for SPC (compared to those not receiving surgery, HR = 0.38, 95% CI: 0.32-0.45, P = 0.001), radiotherapy (compared to those not receiving radiotherapy or unknown, HR = 0.66, 95% CI: 0.56-0.78, P = 0.001), and chemotherapy (compared to those not receiving chemotherapy or unknown, HR = 0.86, 95% CI:0.74-0.99, P = 0.034) were independent protective factors of OS in SPC patients after cervical cancer. The results of Kaplan-Meier survival analysis showed that the differences in the OS of SPC patients after cervical cancer with different marriage status, SEER stage, histologic grade, lymph node dissection, surgery for SPC, primary sites of SPC and whether receiving radiochemotherapy were all statistically significant (all P < 0.001). Based on the 8 variables including marital status, SEER stage, histologic grade, whether lymph nodes have been cleared, whether SPC has been treated with surgery, radiotherapy, chemotherapy, and the primary sites of SPC, a nomogram model for predicting the 1-, 3- and 5-year OS rates of SPC patients after cervical cancer was established. The results of ROC curve analysis showed that the area under the curve of the model for predicting 1-, 3-, and 5-year OS rates was 0.841, 0.847, and 0.847, respectively. The calibration curves showed a good consistency between the predicted results of model and the actual results. Conclusions:A prognostic prediction nomogram model for SPC after cervical cancer constructed based on the data in the SEER database has a high clinical application value and calibration.

Objective:To investigate the role of the Smad3 signaling pathway in the process of silica-induced epithelial-mesenchymal transition (EMT) .Methods:In September 2022, lung epithelial cells (BEAS-2B) were exposed to different concentrations of silica suspension (0, 50, 100, and 150 μg/ml) for 6 and 12 hours. Additionally, SIS3, a specific inhibitor of phosphorylated Smad3 (p-Smad3) , was utilized to establish the p-Smad3 inhibition model. The cells were divided into four groups: blank control gruop, silica group, SIS3 intervention group, and SIS3 +silica group. Cell morphology was observed using an inverted fluorescence microscope, while cell viability was assessed using a Cell Counting Kit-8 (CCK-8) . The mRNA and protein expression levels of E-cadherin (E-Cad) , N-cadherin (N-Cad) , Vimentin, Smad3, and p-Smad3 were analyzed by Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Differences between two groups were compared using Student's t-test, and multiple group comparisons were analyzed using a one-way analysis of variance with the Student-Newman-Keuls test.Results:Compared with the blank control group, the morphology of BEAS-2B cells shifted from epithelial to mesenchymal cell-like following silica exposure, and the cell viability of BEAS-2B cells declined after exposure to 150 μg/ml silica for 6 and 12 hours. Furthermore, silica exposure led to significant reductions in mRNA and protein expression levels of the epithelial cellular marker (E-Cad) in BEAS-2B cells, accompanied by increased expressions of interstitial cellular markers (N-Cad and Vimentin) . Importantly, the level of p-Smad3/Smad3 expression levels was also elevated in silica-treated cells ( P<0.05) . Compared to the blank control group, the level of p-Smad3/Smad3 expression levels was significantty reduced. Moreover, compared to the silica group, the protein expression levels of N-Cad and Vimentin in the cell of the SIS3+silica group were significantly reduced, while the E-Cad expression was increased ( P<0.05) . Conclusion:Silica exposure can prmote the epithelial mesenchymaol transformotion process by activating smod3 signa ling pathuay, and in hibiting smad3 signa ling pathuay can effctively alleviate the occurrence of epithelial mesenchymal transformation process.

Objective:To construct an estimating equation to accurately reflect the aging phenomenon of the glomerular filtration rate(GFR).Methods:Healthy subjects receiving physical examinations at the First Affiliated Hospital of Nanjing Medical University between January 2017 and April 2018 were included in the study, and the aging phenomenon of renal function indicators such as serum creatinine(Scr)was used as the reference standard to evaluate the accuracy of four Scr-based GFR equations during GFR aging, including the full age spectrum(FAS)equation, the Chronic Kidney Disease Epidemiology Collaboration(CKD-EPI)equation, the Osaka equation and the Xiangya equation.Results:Of 37 636 individuals receiving physical examinations, 6 534 met the criteria specified in this study.Scr, serum urea nitrogen, serum uric acid, and serum albumin showed a significant aging phenomenon( H=191.640, 196.693, 83.271, 414.585, P<0.001 for all).The GFR estimated by the four equations all decreased with aging, but the starting point and rate of decline were significantly different.The GFR aging phenomenon estimated by the FAS equation was closer to the trend of renal function indicators. Conclusions:The FAS equation may be more applicable to healthy people to understand the aging phenomenon of GFR.

The modern bio-fermentation industry requires design and creation of efficient microbial cell factories for directed conversion of raw materials to target products. The main criteria for assessing the performance of microbial cell factories are their product synthesis capacity and stability. Due to the deficiencies of plasmids in gene expression such as instability and being easy to lose, integration of genes into chromosome is often a better choice for stable expression in microbial hosts. To this end, chromosomal gene integration technology has received much attention and has developed rapidly. In this review, we summarize the recent research progresses of chromosomal integration of large DNA fragments in microorganisms, illustrate the principles and features of various technologies, highlight the opportunity brought by the CRISPR-associated transposon systems, and prospect future research direction of this technology.
Chromosomes ; Plasmids ; DNA ; Cloning, Molecular ; Fermentation

Objective:To explore the phenotype and molecular genetic features of spinocerebellar ataxia type 2 (SCA2) cases with ATXN2 intermediate-length CAG-repeat expansion.Methods:Fragment analysis by capillary electrophoresis was performed to detect the dynamic mutations in the samples of the probands in 1 383 pedigrees with autosomal dominant inherited ataxia in Research Center for Motor Disorders and Neurogenetic Diseases, Department of Neurology, China-Japan Friendship Hospital from 2005 to 2018. The clinical and genetic features of individuals carrying the ATXN2 intermediate-length CAG-repeat expansion were carefully analyzed.Results:Two hundred and three individuals (including the probands and members of their families) in 163 families carried the expanded CAG repeats in ATXN2 gene, among which 107 individuals in 93 families carried the intermediate-length CAG-repeats. Within 20 parent-child pairs, the CAG repeats increased 0-28 copies in 16 pairs with paternal inheritance, and 0-4 copies in 4 pairs with maternal inheritance.Conclusions:For suspected SCA2 cases, ATXN2 gene testing should be performed on the parental members and adult offspring members in the family. Dynamic mutations testing is essential to identify the individuals with ATXN2 intermediate-length repeat expansion, which is very important for genetic counseling.

Rare diseases have been a major challenge for clinical medicine and public health challenge in China. One of the effective measures is to conduct proactive research on rare diseases to deal with the disease burden of the diseases. However, low prevalence, disperse distribution of patients, lack of knowledge about the disease course, and phenotype heterogeneity hamper the development of research for rare diseases. Recently, it has been found that patients registry is effective in understanding the course of the disease and accu- mulating the cases and data of clinical research or clinical trial design. At present, most of developed countries or regions in the world have promoted clinical research and clinical trials of new medications on rare diseases by using the registration of rare disease. In 2016, Peking Union Medical College Hospital established China's first registry system at the national level-National Rare Disease Registry System of China(NRDRS). NRDRS has accumulated 68 137 cases data registered by the researchers from China's 101 collaborating hospitals in 29 provinces/municipalities/autonomous regions, covering 171 different, and forming 188 cohorts. To date, NRDRS complete the initial stage of resources buildup.Nex stage will be focused on clinical research and clinical trials related to rare diseases based on NRDRS. This article is on the process of building NRDRS, the potential support for conducting clinical research and clinical trials related to rare diseases, and the challenges will be faced.

Objective:To investigate the prevalences of overweight and obesity and its relationship with the risk factors of cardiovascular diseases among physical examination population in Nanjing Drum Tower Hospital.Methods:A retrospective analysis was performed on 384 061 adults aged 20 years and older. Age- and sex-standardized prevalences of overweight and obesity were calculated using Chinese census data in 2010. Multivariate logistic regression analysis was used to assess the associations of overweight and obesity with the risk factors of cardiovascular diseases.Results:The age-standardized prevalences of overweight and obesity were 42.8% and 13.2% in men and 23.9% and 6.6% in women. A gradually increasing trend was observed in the prevalences of overweight and obesity from 2008 to 2016, especially in subjects aged 20-39 years. Overweight and obesity were significantly associated with increased risks of dyslipidemia, diabetes mellitus, hypertension, and hyperuricemia. These associations were found to be the strongest among subjects aged 20-39 years, which became weaker along with the increasing of age. The OR(95% CI) of dyslipidemia, diabetes mellitus, hypertension, and hyperuricemia were 4.23(4.01-4.47), 3.70(2.97-4.60), 6.19(5.76-6.64), and 3.66(3.45-3.88) in obese men aged 20-39 years, while 5.29(4.63-6.04), 6.38(3.86-10.55), 9.36(7.86-11.13), and 6.65(5.70-7.74) in obese women aged 20-39 years, respectively. Conclusion:The increasing trend in the prevalence of overweight and obesity is a risk factor for cardiovascular diseases in Nanjing adults, especially in individuals aged 20-39 years.