Objective:To discuss the effect of long non-coding RNA(lncRNA)DUXAP8 in exosomes(Exo)derived from the lung cancer A549 cells on the growth and immune escape of the lung cancer cells,and to clarify the mechanism.Methods:The human lung cancer cell line A549 was cultured,and its exosomes were extracted and identified.The A549 cells were treated with PKH67-labeled Exo to observe the uptake of Exo by A549 cells.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression level of lncRNA DUXAP8 in A549 cells before and after Exo treatment.The A549 cells were divided into control group(no treatment),Exo group(A549 cells treated with Exo),Exo+sh-NC group(A549 cells treated with Exo and then transfected with sh-NC),and Exo+sh-DUXAP8 group(A549 cells treated with Exo and then transfected with sh-DUXAP8).RT-qPCR method was used to detect the expression level of lncRNA DUXAP8 in A549 cells in various groups;colony formation assay was used to detect the colony formation abilities of the A549 cells in various groups;5-ethynyl-2'-deoxyuridine(EdU)staining method was used to detect the proliferation abilities of the A549 cells in various groups.After co-culturing A549 cells in various groups with human peripheral blood lymphocytes,flow cytometry was used to detect the percentages of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in various groups;3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)method was used to detect the killing rates of human peripheral blood lymphocytes on the A549 cells in various groups.Results:The diameter of Exo vesicles was 50-150 nm,and the exosome-specific marker proteins cluster of differentiation 63(CD63),cluster of differentiation 9(CD9),tumor susceptibility gene 101(TSG101),and heat shock protein 70(HSP70)were positively expressed,indicating successful exosome extraction.A549 cells efficiently took up PKH67-labeled Exo.The RT-PCR results showed that compared with A549 cells cultured alone,the expression level of lncRNA DUXAP8 in the A549 cells was increased after treatment with Exo derived from A549 cells(P<0.05).compared with control group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there were no significant difference in the expression level of IncRNA DUXAP8 in the cells in Exo+sh-NC group(P>0.05).The colony formation assay results showed that compared with control group,the number of colony formation of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the number of colony formation of the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the number of colony formation of the A549 cells in Exo+sh-NC group(P>0.05).The EdU staining results showed that compared with control group,the EdU-positive rate of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the EdU-positive rate in A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the EDU-positive rate in the cells in Exo+sh-NC group(P>0.05).The flow cytometry results showed that compared with control group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo group was decreased(P<0.05);compared with Exo group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo+sh-DUXAP8 group was increased(P<0.05),while there was no significant difference in the percentage of activated CD8+T lymphaytes in Exo+sh-NC group(P>0.05).The MTT assay results showed that compared with control group,the killing rate of human peripheral blood lymphocytes on the A549 cells in Exo group was decreased(P<0.05);compared with Exo group,the killing rate of human peripheral blood lymphocytes on A549 cells in Exo+sh-DUXAP8 group was increased(P<0.05),while no significant difference was observed in Exo+sh-NC group(P>0.05).Conclusion:The lncRNA DUXAP8 in exosomes derived from the lung cancer A549 cells promotes the proliferation of lung cancer cells and tumor immune escape.

Radiotherapy is widely used in the management of advanced colorectal cancer (CRC). However, the clinical efficacy is limited by the safe irradiated dose. Sensitizing tumor cells to radiotherapy via interrupting DNA repair is a promising approach to conquering the limitation. The BRCA1-BARD1 complex has been demonstrated to play a critical role in homologous recombination (HR) DSB repair, and its functions may be affected by HERC2 or BAP1. Accumulated evidence illustrates that the ubiquitination-deubiquitination balance is involved in these processes; however, the precise mechanism for the cross-talk among these proteins in HR repair following radiation hasn't been defined. Through activity-based profiling, we identified PT33 as an active entity for HR repair suppression. Subsequently, we revealed that BAP1 serves as a novel molecular target of PT33 via a CRISPR-based deubiquitinase screen. Mechanistically, pharmacological covalent inhibition of BAP1 with PT33 recruits HERC2 to compete with BARD1 for BRCA1 interaction, interrupting HR repair. Consequently, PT33 treatment can substantially enhance the sensitivity of CRC cells to radiotherapy in vitro and in vivo. Overall, these findings provide a mechanistic basis for PT33-induced HR suppression and may guide an effective strategy to improve therapeutic gain.

Objective:To provide evidence for the selection of fixation devices and CTV to PTV margins (M ptv) in precision radiotherapy for pelvic tumors by analyzing three fixation devices in precision radiotherapy for prostate cancer. Methods:From April 2015 to December 2020, 133 prostate cancer patients treated with pelvic drainage area irradiation in our center were retrospectively analyzed. The patients were fixed with 1.2m vacuum bag (n=39), 1.8m vacuum bag (n=44) and personalized prone plate by our center (n=50). Each patient was asked to complete our bowel and bladder preparation process before positioning and radiotherapy. The registration of CBCT to planned CT before each treatment adopted the same registration box and algorithm. Setup errors in the SI, LR and AP directions under qualified bowel and bladder conditions were recorded. Setup errors in three directions under three fixation devices and corresponding M ptv values were analyzed. The correlation between setup errors with age and body mass index (BMI) was analyzed. Results:Analysis of 3333 setup errors data showed: in the SI and LR directions, the mean setup errors of 1.2m vacuum bag (3.26mm, 2.34mm) were greater than those of 1.8m vacuum bag (2.51mm, P<0.001; 1.90mm, P<0.001), and personalized prone plate (3.07mm, P=0.066; 2.10 mm, P=0.009). In the AP direction, the mean setup errors of 1.2m vacuum bag (supine)(2.20mm) were smaller than those of 1.8m vacuum bag (3.33mm, P<0.001) and personalized prone plate (3.61mm, P<0.001). The setup errors of 1.8m vacuum bag in all directions were smaller than those of personalized prone plate (P≤0.028). According to Van Herk's expansion formula, the M ptv of 1.2m vacuum bag in three directions was approximately 4 mm. The M ptv of 1.8m vacuum bag and personalized prone plate in the SI and LR directions was approximately 3 mm, and more than 5 mm in the AP direction. The setup errors were not correlated with age or BMI. Conclusions:From the setup errors results of three devices, 1.8m vacuum bag is the best, followed by personalized prone plate. And supine position is better than prone position in the AP direction.

BACKGROUND: Pathological complete response (pCR) of axillary lymph nodes (ALNs) is frequently achieved in patients with clinically node-positive breast cancer after neoadjuvant chemotherapy (NAC), and ALN status is an important prognostic factor for breast cancer patients. This study aims to develop a new predictive clinical model to assess the ALN pCR rate after NAC. METHODS: This was a retrospective series of 467 patients who had biopsy-proven positive ALNs at diagnosis and underwent ALN dissection from 2007 to 2014 at the National Cancer Center/Cancer Hospital of the Chinese Academy of Medical Sciences. We analyzed the clinicopathologic features of the patients and developed a nomogram to predict the probability of ALN pCR. A multivariable logistic regression stepwise model was used to construct a nomogram to predict ALN pCR in node-positive patients. The adjusted area under the receiver operating characteristic curve (AUC) was calculated to quantify the ability to rank patients by risk. Internal validation was performed using the 50/50 hold-out validation method. The nomogram was externally validated with prospective cohorts of 167 patients from 2016 to 2018 at the Cancer Hospital of the Chinese Academy of Medical Sciences and 114 patients from 2018 to 2020 at Beijing Tiantan Hospital. RESULTS: In this retrospective study, 115 (24.6%) patients achieved ALN pCR after NAC. Multivariate analysis showed that clinical tumor stage (Odds ratio [OR]: 0.321, 95% confidence interval [CI]: 0.121-0.856; P = 0.023); primary tumor response (OR: 0.189; 95% CI: 0.123-0.292; P < 0.001), and estrogen receptor status (OR: 0.530, 95% CI: 0.304-0.925; P = 0.025) were independent predictors of ALN pCR. The nomogram was constructed based on the result of multivariate analysis. In the internal validation of performance of nomogram, the AUCs for the training and test sets were 0.719 and 0.753, respectively. The nomogram was validated in external cohorts with AUCs of 0.720, which demonstrated good discriminatory power in these data sets. CONCLUSION: We developed a nomogram to predict the likelihood of axillary pCR in node-positive breast cancer patients after NAC. The predictive model performed well in multicenter prospective external validation. This practical tool could provide information to surgeons regarding whether to perform additional ALN dissection after NAC. TRIAL REGISTRATION ChiCTR.org.cn, ChiCTR1800014968.
Breast Neoplasms/drug therapy* ; Female ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Neoadjuvant Therapy ; Nomograms ; Prospective Studies ; Retrospective Studies

Polycyclic aromatic hydrocarbons (PAHs) are widely found in the environment, and comparing to adults, children are more vulnerable to PAHs exposure. Urinary metabolites of PAHs are used as preferred biomarkers to estimate the PAHs exposure. Systematic review on the internal exposure level of children and adolescents is rare. We aimed to calculate the internal exposure levels of PAHs in children and adolescents and compare the levels of PAHs internal exposure in various children groups. We searched PubMed, OVID, Web of Science, EBSCO, ACS, and four Chinese databases, and all studies examining the urinary concentrations of PAHs in children and adolescent were identified. The total exposure level of 11 PAHs metabolites were pooled. Standard mean difference (SMD) and 95% confidence intervals (CIs) of PAHs urinary concentration were calculated and pooled by RevMan5.3 to compare the exposure levels of different children groups. We found that 1-OHPyr, 2-OHNap, 2-OHFlu, 3-OHPhe, and 4-OHPhe were five PAHs metabolites most commonly studied in existing studies in children, and their total exposure levels were 0.38 ± 0.98, 2.32 ± 4.83, 0.81 ± 1.54, 0.09 ± 0.14, 0.03 ± 0.10 μmol/mol creatinine, respectively. The meta-analysis showed that the levels of 1-OHPyr were higher in higher environmental exposure group (SMD = 0.21, 95% CI = 0.03~0.40), ETS exposure group (SMD = 0.31, 95% CI = 0.08~0.54), and 6~11 years group (SMD = 0.16, 95% CI = 0.09~0.24); the level of 2-OHNap (SMD = 0.27, 95% CI = 0.01~0.53) was higher in higher environmental exposure group; however, the levels of 3-OHPhe (SMD = - 0.34, 95% CI = - 0.57~- 0.12) and 4-OHPhe (SMD = - 0.48, 95% CI = - 0.69~- 0.28) were higher in lower environmental exposure group. The levels of 1-OHPyr (SMD = - 0.01, 95% CI = - 0.11~0.10) and 2-OHNap (SMD = 0.01, 95% CI = - 0.20~0.22) were not statistically different between boys and girls. In conclusions, we found that the internal diversity of PAHs existed in children and adolescents, and the level of 1-OHPyr in children and adolescents was in higher status compared with non-occupational people who do not smoke.

Objective To investigate the feasibility and clinical significance of sentinel lymph node biopsy(SLNB) after neoadjuvant chemotherapy (NAC) for axillary lymph node-positive breast cancer.Methods Enrolled for a prospective cohort study were 167 patients from Jan 2016 to Jan 2018 with axillary lymph node-positive breast cancer admitted to the Cancer Hospital of Chinese Academy of Medical Sciences.SLNB was performed after NAC by lymphatic dual mapping,followed by axillary lymph node dissection.The primary end point was sentinel lymph node identification rate (IR) and false negative rate (FNR).Results 62 patients (37.1％) had complete pathological response of axillary lymph nodes.There was a significant difference of NAC response in patients with different subtypes (P ＜0.001).The IR of SLNB after NAC was 94.6％,the FNR was 6.7％,the sensitivity was 93.3％,the specificity was 100％,and the accuracy was 95.8％.Univariate analysis showed that there was no significant difference between tumor stage,hormone receptor status,HER2 expression,and pathological remission in SLN detection group and the SLN undetected group (P ＞ 0.05).The proportion of patients who received breast conserving surgery in the undetected group was significantly higher than that in the test group (P =0.006).Conclusions Sentinel lymph node biopsy after breast neoadjuvant chemotherapy by lymphatic dual mapping is highly accurate with a high identification rate and a low false negative rate.

OBJECTIVES: Mothers who smoke during pregnancy or while their children are small were common in some populations. Epidemiological studies have tried to detect the effect of prenatal tobacco smoke (PTS), and childhood environmental tobacco smoke (ETS) on puberty timing have not shown a consensus results. We aimed to examine current evidence and estimate the associations between PTS or/and ETS and puberty timing. METHODS: Seven databases were searched from inception to May 2017. All the cohort studies examining the associations between PTS and/or ETS and puberty timing were identified. Two reviewers independently screened all studies, evaluated the quality of eligible studies, and extracted the data. The quality assessment of the eligible cohort studies was based on the Newcastle-Ottawa Scale. Risk ratio (RR), standard mean difference (SMD), and 95% confidence intervals (CIs) were calculated and pooled by CMA (Version 2.0, Biostat, Inc., USA). RESULTS: Compared with controls, girls with PTS and ETS exposure have an earlier age at menarche (SMD - 0.087, 95% CI 0.174 to - 0.000), and similar results were found in both PTS subgroup (SMD - 0.097, 95% CI - 0.192 to - 0.002) and prospective cohort subgroup (SMD - 0.171, 95% CI - 0.253 to - 0.090). And number of boys with early voice break in PTS group was significantly increasing than non-exposed boys (RR 1.34, 95% CI 1.29 to 1.40). CONCLUSIONS PTS exposure possibly decrease age of menarche of girls, and studies on boys were urgent needed. Appropriate and comprehensive outcome measures using unified criteria to classify puberty should be reported in future studies.
Aging ; physiology ; Environmental Exposure ; adverse effects ; Female ; Humans ; Menarche ; physiology ; Pregnancy ; Prenatal Exposure Delayed Effects ; etiology ; Puberty ; physiology ; Smoking ; adverse effects ; Tobacco Smoke Pollution ; adverse effects

Objective:To detect the expressions of p16INK4a protein in the cervical lesion tissues of the Mongolian patients, and to explore the relationship between its expression and the occurrence and development of cervical cancer in the Mongolian patients.Methods:A total of 100 cases of paraffin sections of cervical cancer, cervical intraepithelial neoplasia(CIN),chronic cervicitis and uterine leiomyoma were divided into 25 cases of cervical cancer, 35 cases of CIN, 20 cases of chronic cervicitis, and 20 cases of uterine leiomyoma groups. The expressions of p16INK4a protein in different cervical tissues were detected by immunohistochemical SP method.Results:The positive rates of p16INK4a protern in cervical cancer, CIN, chronic cervicitis and uterine leiomyoma tissnes were 100.0%, 74.3%, 25.0%,and 10.0%, respectively.The results of K-W H rank sum test for multiple sample comparisons showed that the positive expression rate of p16INK4a protein in cervical cancer tissue was significantly higher than those in CIN, chronic cervicitis and uterine leiomyoma tissues(P<0.05).Conclusion:p16INK4a protein can be used as a indicator to screen the Mongolian patients with early cervical cancer.

Objective: Breast intraductal papillary tumors are clinically common diseases derived from the ducts. The aim of this study is to investigate the clinicopathological characteristics of intraductal papillary tumors and risk factors for carcinogenesis. Methods: The clinicopathological data of 674 patients with breast intraductal papillary tumors, who underwent surgery in the National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences from January 2010 to July 2015, were retrospectively analyzed. Results: The median follow-up time was 46 months. The 674 cases were classified into 547 intraductal papilloma, 88 cases of intraductal papillary carcinoma, 32 cases of intracystic papillary carcinoma, and 7 cases of solid papillary carcinoma of breast. After a median follow-up time of 46 months, 13 out of 547 (2.4%) intraductal papillomas had local recurrence in the original dissected quadrat, another 10 cases developed breast cancer in the original dissected quadrat. The 3-year recurrence-free survival rates in intraductal papilloma and intraductal papilloma accompanied with atypical ductal hyperplasia were 97.7% and 93.5%, respectively, the recurrence-free survival curves showed a significant difference (P=0.011). Multivariate analysis indicated that atypical ductal hyperplasia was a major prognostic factor affecting the recurrence-free survival of intraductal papilloma (RR=0.183, 95%CI=0.054 to 0.777, P=0.020). Four cases (3.1%) of intraductal papillary carcinoma had local recurrence. The logistic analysis showed that patient aged >45 years, clinical manifestations of a breast lump, maximum tumor diameter greater than 2 cm are possible clinical manifestation of malignant breast intraductal papillary tumors (RR=1.735, 95%CI=1.007-2.990, P=0.047; RR=2.849, 95%CI=1.207-6.711, P=0.017; RR=3.792, 95%CI=2.162-6.653, P<0.001). Conclusions Intraductal papillary tumors have a certain recurrence rate. Age, clinical features and tumor size may be predictive factors of intraductal papillary carcinoma.

Objective To explore the impact of environment factors and social economic factors(environment interference factors for short)and control measures on the endemic status of schistosomiasis in Poyang Lake Eco?economic Region. Meth?ods The grey relational analysis model was applied to analyze the relationships between the key indexes of schistosomiasis epi?demic status and the environment interference factors and control measures in Jiujiang City of Poyang Lake Eco?economic Re?gion. Results Six environment factors,which included the annual average water level of Poyang Lake,average annual tempera?ture,storm frequency,annual average relative humidity,annual sunshine duration,and annual precipitation,had the most closed relationship with the indexes of schistosomiasis epidemic status(all ri>0.9). Among the socioeconomic factors,the num?ber of health technicians and beds of health facilities were most associated with the indexes of schistosomiasis epidemic status. Among the control measures of schistosomiasis,the number of cattle treated with extending chemotherapy,chemically killing of Oncomelania hupensis snails and eco?renovation were most associated with the indexes of schistosomiasis epidemic status. Con?clusion The various environment interference factors and their interaction should be considered in formulating the comprehen?sive control strategy for schistosomiasis,and the control strategy should be adjusted according to the epidemic dynamic and schistosomiasis?focused targets,so as to further strengthen the scientificity and validity of the control strategy.