Purpose: Oligometastatic non–small cell lung cancer (NSCLC) patients have been increasingly regarded as a distinct group that could benefit from local treatment to achieve a better clinical outcome. However, current definitions of oligometastasis are solely numerical, which are imprecise because of ignoring the biological heterogeneity caused by genomic characteristics. Our study aimed to profile the molecular alterations of oligometastatic NSCLC and elucidate its potential difference from polymetastasis. Materials and Methods: We performed next-generation sequencing to analyze tumors and paired peripheral blood from 77 oligometastatic and 21 polymetastatic NSCLC patients to reveal their genomic characteristics and assess the genetic heterogeneity. Results: We found ERBB2, ALK, MLL4, PIK3CB, and TOP2A were mutated at a significantly lower frequency in oligometastasis compared with polymetastasis. EGFR and KEAP1 alterations were mutually exclusive in oligometastatic group. More importantly, oligometastasis has a unique significant enrichment of apoptosis signaling pathway. In contrast to polymetastasis, a highly enriched COSMIC signature 4 and a special mutational process, COSMIC signature 14, were observed in the oligometastatic cohort. According to OncoKB database, 74.03% of oligometastatic NSCLC patients harbored at least one actionable alteration. The median tumor mutation burden of oligometastasis was 5.00 mutations/Mb, which was significantly associated with smoking, DNA damage repair genes, TP53 mutation, SMARCA4 mutation, LRP1B mutation, ABL1 mutation. Conclusion Our results shall help redefine oligometastasis beyond simple lesion enumeration that will ultimately improve the selection of patients with real oligometastatic state and optimize personalized cancer therapy for oligometastatic NSCLC.

Objective:To improve the critical thinking ability of interns in nephrology department based on electronic "Spot" mind mapping teaching method.Methods:In the control group, the traditional clinical teaching method was adopted. Each kidney disease unit was divided into 3 courses. ①The clinical practice teacher dictated or demonstrated his/her experience to the student in the first class. ②Students could exchange questions and answers in the second class. ③In the third class, according to the homework situation, the teacher presided over the discussion, guided the students to express their difficulties and help them solve the problems. The research group adopted the electronic "Spot" mind mapping teaching method: ①Grouping: the students were divided into groups, 6 to 8 people in each group, a total of 16 groups. ②Preparation: each group established a WeChat group, and teachers guided them download the Mindmanager software and learn its mapping method. ③In class: each kidney disease unit was divided into 3 sessions. In the first class, based on what the instructor taught, the students summarized the contents and drew a mind map, and then explain their understanding according to the map. In the second class, "Spot" in the group was conducted based on standards, reading each other in the group, actively discussing with each other, further improving and reconstructing the core knowledge points of the chapter, and encouraging each student to actively participate in enhancing their subjective initiative in learning. In the third class, teachers evaluated students according to their learning situation, and students filled in the gaps according to their opinions, perfected their mind maps, and finally posted to the WeChat group. ④Review: the final versions were sent to the WeChat groups as review materials, which was convenient for learning together. SPSS 24.0 was used for Chi-square test.Results:There was no significant difference between the two groups before study ( P>0.005). After the research, the scores of theory test ( t=2.52, P=0.015), clinical skill test ( t=2.22, P=0.034) and total score ( t=3.53, P=0.003) in the experimental group were significantly higher than those in the control group ( P<0.05). There was no significant difference in the scores of critical thinking ability between the two groups before research ( P>0.05). Six months after research, the total scores of critical thinking ability in the experimental group were significantly higher than those in the control group ( P<0.05). Conclusion:The introduction of electronic "Spot" mind mapping teaching method into clinical practice teaching can realize the cross-linking of related knowledge points and systematize the knowledge. At the same time, it is interesting and can stimulate students' learning interest, and is helpful to cultivate the clinical critical thinking ability of students.

Objective:To investigate the role of miRNA-4298/PADI4/p53 signal axis in mediating 4f-induced apoptosis of leukemia cells.Methods:The cell growth density was observed under inverted microscope and the proliferation of leukemia cells was detected by CCK-8 counting assay. The expression of PADI4 and P53 at mRNA level was detected by qRT-PCR. Cell cycle and apoptosis were measured with flow cytometry. The expression of PADI4, P53, Bcl-2, Bax, caspase-3 and caspase-9 at protein level was detected by Western blot. Differential miRNA and mRNA expression profiles was detected by next generation sequencing. Databases such as TargetScan were used to predict the potential upstream and downstream genes of PADI4. A luciferase reporter assay was used to detect the 3′UTR of PADI4 targeted by miRNA-4298. Cell transfection assay was used to detect the effect siRNA, PADI4 vector, miRNA mimics and miRNA inhibitor in interference and rescue.Results:Nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor 4f could inhibit the proliferation of THP-1, K562 and U937 cells, and induce the apoptosis of these leukemia cells. It downregulated the expression of PADI4 mainly through the binding activity of miRNA-4298 to miRNA sponges, which resulted in the proliferation inhibition and apoptosis of leukemia cells. The inhibited proliferation and apoptosis of leukemia cells by 4f were associated with the increase of P53 expression after the decrease of PADI4 expression. The PADI4-dependent upregulation of P53 led to the ratio inversion of downstream Bcl-2/Bax, which activated caspase-3 or caspase-9 to induce the apoptosis of leukemia cells.Conclusions:The apoptosis of leukemia cells induced by Nrf2 inhibitor 4f was mainly associated with the miRNA-4298/PADI4/p53 axis, suggesting that it might be a novel signaling pathway for targeted therapy.

Objective:To detect serum levels of vitamin A (Vit A), vitamin D(Vit D)25-hydroxy vitamin D[25-(OH)D] and vitamin E(Vit E) in children aged 0-6 years in Tibetan Plateau of Garzi Prefecture, thus providing references for physical examinations and prevention of 4 key diseases (rickets, malnutrition anemia, pneumonia and diarrhea) in children in plateau areas by relevant government departments.Methods:A total of 2 122 children who participated in physical examination in 12 townships of Xiangcheng County and 14 townships of Daocheng County, Garzi Tibetan Autonomous Prefecture, Sichuan Province from April 2017 to April 2019 with 0-6 years old were recruited for surveying physical measurements and collection of venous blood.Serum Vit A and Vit E levels were detected by high performance liquid chromatography.Serum levels of 25-(OH)D were detected by high performance liquid chromatography tandem mass spectrometry.The relationship between Vit A, Vit E and 25-(OH)D levels with the gender, age, seasonal change and altitude was analyzed.Results:The serum Vit A level, subclinical Vit A deficiency rate and marginal vitamin A deficiency rate were(1.05±0.27) μmol/L, 8.15%(173/2 122 cases) and 45.99%(976/2 122 cases), respectively in 2 122 children with 0-6 years old.There were significant differences in the serum Vit A level, the subclinical Vit A deficiency rate and the marginal vitamin A deficiency rate in children with different ages, seasons and altitudes (all P<0.05). The serum level of 25-(OH)D and 25-(OH)D deficiency rate insufficient rate were (24.65±6.45) ng/L, 6.03%(128/2 122 cases) and 16.59%(352/2 122 cases), respectively.There were significant differences in the serum level of 25-(OH)D, 25-(OH)D deficiency rate and 25-(OH)D insufficient rate in children with different ages and seasons (all P<0.05). The mean serum Vit E level, Vit E deficiency rate and Vit E insufficient rate were (7.81±1.74) mg/L, 2.78%(59/2 122 cases) and 29.59%(628/2 122 cases), respectively.There were significant differences in serum Vit E level, Vit E deficiency rate and Vit E insufficient rate in children with different ages and seasons (all P<0.05). The mean serum levels of Vit A and Vit D remained the lowest before the age of 1 year, and their deficiencies at this age were the most significant.The mean serum level of Vit E remained the lowest in >1-2 years old, and its deficiency and insufficient at this age were the most significant.Vit A, D and E levels were significantly affected by seasonal changes, which were significantly higher in the summer than in the spring, autumn and winter.In addition, Vit A and 25-(OH)D were significantly affected by the altitude, which were the lowest above 4 km altitude. Conclusions:The overall serum levels of Vit A, 25-(OH) D and E in children with 0-6 years old in Tibetan Plateau areas of Ganzi Prefecture are lower than those in plain areas.Vit A, 25-(OH) D and Vit E levels significantly differed in the age, season and altitude, which are related to the lack of local resources, insufficient maternal nutrition during pregnancy and insufficient intake after birth, as well as temperature and light caused by changes in local seasons and altitude.Therefore, it is necessary to make reasonable supplements during pregnancy to prevent vitamin deficiency.

Background and purpose:The young breast cancer patients were treated with goserelin without individualized regimen, and lack of available clinical marker. The aim of this study was to investigate the role of anti-Müllerian hormone (AMH) in evaluation of individualized treatment of ovarian function suppression in the young breast cancer patients.Methods:Forty-one young patients with estrogen receptor (ER) and progesterone receptor (PR) positive breast cancer from May 2012 to Jan. 2014 were randomly divided into 2 groups to undergo radical resection of breast cancer. According to postoperative treatment, one group was treated with goserelin + chemotherapy (n=20), and the other group received chemotherapy alone (n=21). Thirty female patients in the same age group were selected as normal control group. The time of menopause and menstrual recovery after the goserelin + chemotherapy or chemotherapy alone were observed in 2 groups. In early follicular phase (day 3-5) of the cycle preceding the operation and 3, 6 courses after the goserelin + chemotherapy treatment or chemotherapy treatment, serum levels of AMH, FSH and E2 were measured in 2 groups. Accordingly, serum levels of AMH, FSH and E2 were evaluated as well in normal control group.Results:There were no signiifcant differences in preoperative general conditions and preoperative serum FSH and E2 levels among the 3 groups (P>0.05). Compared with normal control group, the preoperative serum AMH levels of young breast cancer patients were decreased signiifcantly (P=0.04). The menopause time and menstrual recovery time in 2 chemotherapy groups were signiifcantly shorter than that in normal control group (P=0.00). Compared with normal control group and preoperative measurement, the differences in serum FSH and E2 levels were not statistically significant in goserelin + chemotherapy group or chemotherapy alone group (P<0.05). The serum AMH levels measured at different time points of the goserelin + chemotherapy group and chemotherapy alone group were decreased signiifcantly (P<0.05). Compared with the chemotherapy group, the serum AMH levels of the goserelin + chemotherapy group after 6 courses were signiifcantly decreased, and then signiifcantly increased 6 months after menstrual recovery (P<0.05).Conclusion:This study demonstrated that the serum AMH levels were obviously decreased after the ovarian function suppression treatment and increased after the menstrual recovery compared with evaluation of other ovarian reserve index. The serum AMH level could suggest ovarian reserve damage even after ovarian function has recovered to the noticeable level. Thus, AMH could be used clinically to evaluate the ovarian reserve of breast cancer patients as a potential marker for the individualized ovarian function suppression treatment in young breast cancer patients.

OBJECTIVETo investigate Runx3 and C-myc expressions in colorectal cancer and their relationship with the clinicopathological parameters.

METHODSReal-time quantitative PCR was used to detect Runx3 and C-myc mRNA expressions in 38 colorectal cancer tissues and matched adjacent tissues, and Runx3 and C-myc expressions was detected by Western blotting in 63 pairs of colorectal cancer and adjacent tissues. The results were stratified according to the clinicopathological characteristics to examine the relationship of Runx3 and C-myc expressions with the clinicopathological factors in the patients.

RESULTSRunx3 expression was down-regulated and C-myc expression up-regulated at both mRNA and protein levels in colorectal cancer tissues compared with the normal tissues, and their protein expressions exhibited an inverse correlation (r=-0.398, P=0.001). Runx3 and C-myc expressions differed significantly between tumors with different Dukes stages, depths of tumor invasion, lymph node statuses, or histological differentiation (P<0.05); Runx3 down-regulation and C-myc up-regulation were more obvious in tumors in advanced Dukes stage and in poorly differentiated tumors.

CONCLUSIONAbnormal expressions in Runx3 and C-myc may contribute to the occurrence and development of colorectal cancer and are closed correlated with the patient's clinicopathological parameters.

Blotting, Western ; Cell Differentiation ; Colorectal Neoplasms ; genetics ; metabolism ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; Down-Regulation ; Humans ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Up-Regulation

OBJECTIVETo detect the methylation status of TMS1 gene and its demethylation by arsenic trioxide (As₂O₃) in K562 cells.

METHODSK562 cells were treated with different concentrations of As₂O₃ for 48 hours. Methylation-specific PCR (MSP) was used to determine the methylation status of TMS1. RT-PCR and Western blot were used to detect the levels of TMS1 mRNA and protein. TMS1 associated apoptosis proteins Bcl-2/Bax were also analyzed by Western blot. Apoptosis were evaluated by flow cytometry using Annexin V/propium iodide (PI) double staining.

RESULTSTMS1 gene was completely methylated in K562 cells and the levels of TMS1 mRNA and protein were low (0.01±0.01, 0.09±0.02), which could be reversed (mRNA: 0.72±0.04; protein: 1.30±0.06; P<0.01) by 2 μmol/L As2O3 via overt demethylation of TMS1 gene. Apoptosis in experiment group (12.24±1.06) was significantly higher than that in control group (2.05±0.16, P<0.05). In experiment group, the down-expression of antiapoptotic protein Bcl-2 and up-expression of pro-apoptotic protein Bax led to an obvious decline ratio of Bcl-2/Bax (0.56±0.12), as compared to the control group (1.94±0.14, P<0.01).

CONCLUSIONAs₂O₃ could up-regulate TMS1 gene expression by reversing its hypermethylation and induced apoptosis by down-regulation of Bcl-2/Bax ratio in K562 cells.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; CARD Signaling Adaptor Proteins ; Cytoskeletal Proteins ; metabolism ; DNA Methylation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Oxides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To investigate Runx3 and C-myc expressions in colorectal cancer and their relationship with the clinicopathological parameters. Methods Real-time quantitative PCR was used to detect Runx3 and C-myc mRNA expressions in 38 colorectal cancer tissues and matched adjacent tissues, and Runx3 and C-myc expressions was detected by Western blotting in 63 pairs of colorectal cancer and adjacent tissues. The results were stratified according to the clinicopathological characteristics to examine the relationship of Runx3 and C-myc expressions with the clinicopathological factors in the patients. Results Runx3 expression was down-regulated and C-myc expression up-regulated at both mRNA and protein levels in colorectal cancer tissues compared with the normal tissues, and their protein expressions exhibited an inverse correlation (r=-0.398, P=0.001). Runx3 and C-myc expressions differed significantly between tumors with different Dukes stages, depths of tumor invasion, lymph node statuses, or histological differentiation (P<0.05);Runx3 down-regulation and C-myc up-regulation were more obvious in tumors in advanced Dukes stage and in poorly differentiated tumors. Conclusion Abnormal expressions in Runx3 and C-myc may contribute to the occurrence and development of colorectal cancer and are closed correlated with the patient's clinicopathological parameters.

Objective To investigate Runx3 and C-myc expressions in colorectal cancer and their relationship with the clinicopathological parameters. Methods Real-time quantitative PCR was used to detect Runx3 and C-myc mRNA expressions in 38 colorectal cancer tissues and matched adjacent tissues, and Runx3 and C-myc expressions was detected by Western blotting in 63 pairs of colorectal cancer and adjacent tissues. The results were stratified according to the clinicopathological characteristics to examine the relationship of Runx3 and C-myc expressions with the clinicopathological factors in the patients. Results Runx3 expression was down-regulated and C-myc expression up-regulated at both mRNA and protein levels in colorectal cancer tissues compared with the normal tissues, and their protein expressions exhibited an inverse correlation (r=-0.398, P=0.001). Runx3 and C-myc expressions differed significantly between tumors with different Dukes stages, depths of tumor invasion, lymph node statuses, or histological differentiation (P<0.05);Runx3 down-regulation and C-myc up-regulation were more obvious in tumors in advanced Dukes stage and in poorly differentiated tumors. Conclusion Abnormal expressions in Runx3 and C-myc may contribute to the occurrence and development of colorectal cancer and are closed correlated with the patient's clinicopathological parameters.

Objective To investigate the expressions of Survivin and high risk-human papillomavirus (HR-HPV) in cervical carcinoma and premalignant lesion,and explore their roles in the pathogenesis of cervical carcinoma.MethodsEighty-two patients of cervical intraepithelial neoplasia(CIN)and cervical carcinoma were enrolled,including 25 CIN Ⅰ,23 CIN Ⅱ -Ⅲ and 34 cervical carcinoma.Twenty normal cervicals were chosen as control.The expression of Survivin was examined by immunohistochemical assay(SP),and the infection of HR-HPV was measured by polymerase chain reaction (PCR).ResultsExpressions of Survivin and HR-HPV of patients with cervical carcinoma [ 85.3％(29/34)and 88.2％ (30/34) ]were higher than those in CIN [ 52.1％ (25/48) and 54.2％ (26/48) ](P ＜ 0.05 ),CIN were higher than those in normal cervicals [ 0 and 10.0％ ( 2/20 ) ](P ＜ 0.05 ),CIN Ⅱ - Ⅲ [ 65.2％ ( 15/23 ) and 73.9％ ( 17/23 ) ]were higher than those in CIN Ⅰ [ 32.0％ ( 8/25 ) and 28.0％ (7/25) ](P ＜ 0.05 ).Survivin positive expression in cervical carcinoma was correlation with histological grade,but was no correlation with (P ＜ 0.05),age,clinical stage and pathology typing (P ＞ 0.05 ).HR-HPV infection was no correlation with age,clinical stage,histological grade and pathology typing (P ＞ 0.05).Positive correlation between the Survivin positive expression and HR-HPV infection was observed in cervical carcinoma(r =0.403,P ＜0.05).ConclusionIt suggests that Survivin may play an important role in the occurrence and development of cervical carcinoma together with HR-HPV.