OBJECTIVES: To explore the molecular mechanism by which lncRNA SNHG15 regulates proliferation, invasion and migration of lung adenocarcinoma cells. METHODS: The lncRNA microarray chip dataset GSE196584 and LncBase were used to predict the lncRNAs that interact with miR-30b-3p, and their association with patient prognosis were investigated using online databases, after which lncRNA nucleolar RNA host gene 15 (SNHG15) was selected for further analysis. The subcellular localization of lncRNA SNHG15 and its expression levels in normal human lung epithelial cells and lung adenocarcinoma cell lines were detected using fluorescence in situ hybridization and qRT-PCR. In cultured A549 cells, the changes in cell proliferation, migration, and invasion following transfection with a SNHG15 knockdown plasmid (sh-SNHG15), a miR-30b-3p inhibitor, or their co-transfection were assessed with EdU, wound healing, and Transwell assays. Bioinformatics analyses were used to predict the regulatory relationship between lncRNA SNHG15 and COX6B1, and the results were verified using Western blotting and rescue experiments in A549 cells transfected with sh-SNHG15, a COX6B1-overexpressing plasmid, or both. RESULTS: LncRNA SNHG15 was shown to target miR-30b-3p, and the former was highly expressed in lung adenocarcinoma, and associated with a poor patient prognosis. LncRNA SNHG15 was localized in the cytoplasm and expressed at higher levels in A549 and NCI-H1299 cells than in BEAS-2B cells. In A549 cells, lncRNA SNHG15 knockdown significantly inhibited cell migration, invasion and proliferation, and these changes were reversed by miR-30b-3p inhibitor. A regulatory relationship was found between lncRNA SNHG15 and COX6B1, and their expression levels were positively correlated (r=0.128, P=0.003). MiR-30b-3p knockdown obviously decreased COX6B1 expression in A549 cells, and COX6B1 overexpression rescued the cells from the inhibitory effects of lncRNA-SNHG15 knockdown. CONCLUSIONS LncRNA SNHG15 may compete with COX6B1 to bind miR-30b-3p through a ceRNA mechanism to affect proliferation, migration, and invasion of lung adenocarcinoma cells.
Humans ; MicroRNAs/metabolism* ; RNA, Long Noncoding/genetics* ; Cell Proliferation ; Cell Movement ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung ; Neoplasm Invasiveness ; A549 Cells ; Adenocarcinoma/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common type of non-small cell lung cancer, and any change of miRNAs expression will affect the degree of target regulation, thus affecting intracellular homeostasis. This study verified that miR-186-5p could inhibit the proliferation, migration and invasion of LUAD cells by regulating PRKAA2. METHODS: Previous investigations found that the expression of miR-186-5p was markedly suppressed in LUAD. Bioinformatics method is used to predict the target protein related to ferroptosis downstream and inquire about its expression level in LUAD and its influence on the survival of patients. Double luciferase verified the binding site of PRKAA2 and miR-186-5p. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of PRKAA2. The effects of miR-186-5p of LUAD cells as well as the mechanism by which miR-186-5p inhibits Fer-1's sensitivity to ferroptosis were confirmed by EdU, Transwell, and scratch assays. The effect of miR-186-5p on the amount of reactive oxygen species (ROS) in LUAD cells was discovered using ROS experiment. Malondialdehyde (MDA) and glutathione (GSH) experiments were used to detect the effects of miR-186-5p and PRKAA2 on ferroptosis index of LUAD cells. The concentration of lipid ROS (L-ROS) in LUAD cells were measured using the L-ROS tests to determine the effects of miR-186-5p and PRKAA2. RESULTS: The expression of PRKAA2 is up-regulated, and a high level of PRKAA2 expression was associated with a poor prognosis for patients with LUAD. Overexpression of miR-186-5p decreased the gene and protein expression of PRKAA2. By promoting ferroptosis, miR-186-5p overexpression prevented lung cancer cells from proliferating, invading, and migrating. ROS could be produced in higher amounts in LUAD cells due to miR-186-5p. Overexpression of miR-186-5p and knockdown PRKAA2 up-regulated MDA content and reduced GSH content in LUAD cells, respectively. miR-186-5p could increase the content of L-ROS and promote the ferroptosis sensitivity of LUAD cells by targeting PRKAA2. CONCLUSIONS miR-186-5p promotes ferroptosis of LUAD cells through targeted regulation of PRKAA2, thus inhibiting the proliferation, invasion and migration of LUAD.
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Humans ; Lung Neoplasms/genetics* ; Carcinoma, Non-Small-Cell Lung ; Ferroptosis/genetics* ; Reactive Oxygen Species ; Adenocarcinoma of Lung/genetics* ; MicroRNAs/genetics* ; 3,4-Methylenedioxyamphetamine ; Cell Proliferation/genetics* ; Cell Movement/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor ; AMP-Activated Protein Kinases

Objective:To analysis the policy tools and targets of the policies of free training for order-oriented medical students in rural areas of China, for reference for further improving the free medical student training policy.Methods:The research team searched the official websites of the State Council, National Health Commission, Ministry of Education, and other ministries, as well as the Peking University Treasure Database, for national level policy documents related to free training of order-oriented medical students released from June 2010 to May 2023. Policy tool-policy target analysis framework was used to quantitative analysis the policy documents.Results:A total of 16 policy documents were included and 213 policy provisions were extracted. From the perspective of policy tools, the proportion of policy provisions using imperative policy tools was the highest, accounting for 63.38% (135 articles), followed by advisory policy tools(18.78%, 40 articles)and reward based policy tools(13.61%, 29 articles), while functional expansion tools(2.82%, 6 articles) and authoritative restructuring tools(1.41%, 3 articles) accounted for a relatively low proportion. The institutional education stage is the main policy target, with provisions accounting for 76.06% (162 articles), followed by the continuing education stage and the post graduation education stage, accounting for 17.84% (38 articles) and 7.51% (16 articles), respectively. It was uneven distribution of various policy tools and their sub tools within the same policy target.Conclusions:The distribution of policy tools for the free training policy of rural order oriented medical students in China needed to be further balanced. The policy targets were mainly concentrated in the education stage of universities.

Objective:To explore the relationship between executive function and gait in cases of mild amnestic cognitive impairment (aMCI).Methods:Twenty aMCI hospital patients formed an observation group, while 20 healthy counterparts were the control group. Both groups underwent the Tinetti test, followed by the " normal walking" single-task test and the " normal walking + Go/No-go" dual-task test. The pace, step width, stride length, Go/No-go task response time and accuracy rate were recorded.Results:In the single-task test, there was no significant difference in pace or stride width between the two groups, but the average stride length of the observation group (1.11±0.04)cm was significantly shorter than that of the control group. However, in the dual-task test, the average pace time (0.96±0.08)sec and stride length (1.02±0.06)cm of the observation group were significantly smaller than the control group′s averages, while their step width (0.11±0.02)cm was significantly wider. There was no significant difference between the two groups in the response time in a single (Go/No-go) task, but in the dual-task test, the observation group′s average time was significantly longer than the control group′s and the accuracy was significantly poorer. Both the error rate and the non-response rate were significantly higher than among the control group.Conclusions:Mild amnestic cognitive impairment reduces stride length and pace when walking and impairs executive function.

Objective:To summarize the clinical features, laboratory examination and genetic analysis of a patient with mucopolysaccharidosis type Ⅱ (MPS Ⅱ).Methods:Clinical manifestations, results of urine glycosaminoglycans (GAGs) and dermatan sulfate assay, metabolites related to MPS in peripheral blood leukocytes were analyzed. Meanwhile, the child and his mother were subjected to next-generation sequencing and Sanger sequencing.Results:The boy has presented with global development delay, coarse facies, frequent upper-respiratory infections, hearing loss, indirect inguinal hernia, hepatosplenomegaly, and skeletal deformities. His urine GAGs were significantly elevated, and the urinary dermatan sulfate (DS) was positive. Meanwhile, the activity of idose-2-sulfatase was extremely reduced. The patient was found to harbor a hemizygote c. 676C>G (p.His226Asp) missense variant in exon 5 of IDS gene, for which his mother was heterozygous. Conclusion:The novel c. 676C>G variant of the IDS gene probably underlay the MPS Ⅱ in this child. Genetic testing combined with enzymatic analysis can enable effective diagnosis and classification of MPS.

Objective:To explore the experience of the treatment of patella inferior pole fracture with non excitation tension band technique.Methods:From April 2009 to December 2019, 76 patients with inferior patellar fracture were treated with non excitation tension band technique. There were 42 males and 34 females; The age was 35.48 ± 18.12 years (17-66 years), with an average of 35 years. There were 45 cases on the left and 31 cases on the right. Causes of injury: fall injury in 46 cases, bicycle injury in 16 cases and traffic accident injury in 14 cases. The time from injury to operation was 0.25-5 d, with an average of 3.2 d. There were 18 cases of open fractures, 64 cases of articular surface fractures and 52 cases of comminuted fractures. Open fracture patients were debridement first, and all cases were fixed with non excitation tension band technique. The intraoperative fracture reduction, postoperative fracture healing and internal fixation were observed. The knee function was evaluated according to the degree of claudication, the use of supports, the feeling of joint strangulation, joint instability, pain, swelling, stair climbing and squatting.Results:All 76 patients successfully completed the operation and were followed up for 18.24 ± 6.18 months (range 12-24 months). There was no wire fracture during and after operation. All incisions healed in the first stage after operation, and there was no deep venous thrombosis of lower limbs. The X-ray film showed that there was no further fracture and fracture displacement after operation, and there was no Kirschner wire slip before the internal fixation was taken out. Only one case had steel wire relaxation due to buckle (which did not affect fracture healing). All fractures healed well, with an average of 7.5 weeks, and the fracture line basically disappeared. 12 months after operation, according to Lysholm knee score standard, the knee function of patients was evaluated. The score of 76 cases was 95.40±4.60 points (range 82-100 points): excellent in 54 cases, good in 21 cases and fair in 1 case. The excellent and good rate was 98.7%. According to the visual analogue scale (VAS) pain score standard, the patients were scored for postoperative pain. The score of 76 cases was 0.09±2.05 points (range 0-3 points): 4 cases had mild pain, with an average score of 0.09.Conclusion:The results of the treatment of patellar inferior pole fracture with tension free band technique are satisfactory, the pain caused by tissue irritation by the internal fixation was lower, clinical application can be popularized.

【Objective】 To explore and evaluate infection control measures of preventing cross-contamination of novel coronavirus during gastrointestinal endoscopy treatment. 【Methods】 According to the hospital’s infection control requirements and related documents, infection control measures were formulated and implemented by combining with our actual clinical situation, including the management of the endoscope room, management and protection of patients and endoscopists. Then, we evaluated the effect of these measures. 【Results】 From January 25 to March 10, 2020, a total of 71 patients (53 males and 18 females) completed gastrointestinal endoscopy treatment, with an average age of 54 years (28-81 years). There were 36 (50.7%) cases of emergency treatment. All patients had been kept in quarantine for about 14 days (24±13), and no cross-contamination of novel coronavirus occurred. 【Conclusion】 During the novel coronavirus infection epidemic period, reasonable and effective measures should be taken to minimize the risk of infection in doctors and patients. The endoscope center should strengthen preoperative screening and management of patients, master indications of endoscopic procedures, complete endoscopists’ management and protection work, strictly follow the specifications of sterilizing gastrointestinal endoscopes, and construct the layout of "three zones and two passages".

Objective: To explore the association between screen time and autistic behavior in infants and young children, and to provide clues to the mechanism for further research. Methods: The primary caregivers of 22 586 children in the district of Longhua in Shenzhen were surveyed. Demographic data and screen time were collected using a selfdisigned questionnaire, and children’s autistic behavior was assessed using the child Autism Behavior Checklist(ABC).The chisquare test was used to analyze the correlation between demographic data and screen time, demographic data and autistic behavior, and screen time and autistic behavior. Unconditioned Logistic regression model was used to study the effect of electronic screen exposure on autistic behavior. Results: The amount of screen time spent in infancy increased with age. For children ages 0-,1- and 2-3 years, 60.1%, 35.0% and 20.2% respectively did not watch TV, and 74.0%, 52.6% and 26.8% respectively did not watch the new generation of electronic products. The positive rate of ABC scale screening was 5.3%, including 6.0% male and 4.4% female, OR(95%CI)=1.37 (1.23-1.54).There was correlations between screen time and autistic behavior in infants at all ages (P<0.05).Screen time increased the risk of autistic behavior in younger age groups than in older age groups. For 1 year olds with moderate screen exposure, increased screen exposure at 2 to 3 years of age was associated with an increased risk of autistic behaviors, while reduced screen exposure at 2 to 3 years of age was associated with a lower risk of autistic behaviors(OR=2.14, 2.77, P<0.05). The higher daily screen time at 0-3 years old was, the greater risk of autistic behaviors. Compared with the noncontact electronic screen group, the OR values of the daily TV screens in the ≥1 h/d group and the ＜1 h/d group were 2.01 and 2.45, respectively (P＜0.05).Compared with the non-contact electronic screen group, the OR values of the screens exposed to the new generation of electronic products in the ≥1 h/d group and the ＜1 h/d group were 2.01 and 2.33, respectively(P<0.05).The higher the time of single exposure to electronic screen between 0 and 3 years old, the greater the risk of autistic behaviors. The OR values were 2.50, 1.79 and 1.47 when ≥1 h/time, 0.5-1 h/time and 15-30 min/time compared with <15 min/time（P<0.05）. Conclusion Early exposure to electronic screens in infants, excessive total daily exposure to electronic screens, and excessive screen time each time are all likely to increase autistic behavior. Therefore, it is suggested that children under 2 years old should not be exposed to electronic screens every day. Children aged 2-3 years old who are exposed to electronic screens＜0.5 h/d and whose screen time <15 min might not significantly increase autistic behaviors.

Objective:To investigate the clinicopathologic features, diagnosis and differential diagnosis of large nested variant of urothelial carcinoma (LNUC) of urinary bladder.Methods:The clinical and pathologic data of five cases of LNUC of urinary bladder diagnosed between January 2014 and December 2018 at the Department of Pathology, Zhejiang Provincial People′s Hospital were analyzed by immunohistochemistry (IHC) and Sanger sequencing. The relevant literature was reviewed.Results:All five patients were male with a mean age of 63 years (range, 48 to 81 years). The mean tumor size was 3.4 cm (range, 1.7 to 4.7 cm). Histologically, the invasive LNUC tumor cells formed medium to large sized nests of varying shapes, from regular round, bulbous, oval to irregularly fused, branched, dumbbell shaped glands, with mild stromal reaction. In all five cases, focal central necrosis and microcystic changes in the tumor nests were identified. Cytologically, the tumor cells were low grade in four cases; the remaining case was overall low grade with focal high grade areas. Mitoses were scarce. All cases possessed surface urothelial tumors, including three low-grade papillary carcinomas, one high-grade papillary carcinoma and one carcinoma in situ. Three of the LNUC were accompanied by small nested variant of urothelial carcinoma and two by conventional high grade invasive urothelial carcinoma. Perineural involvement and angiolymphatic invasion were each noted in four tumors. Radical cystectomy was performed in four cases with TNM stages as followings: pT3aN0M0 in two cases, pT4aN0M0 and pT4aN1M0 in one case each. The remaining case had transurethral bladder resection and was of pT2 stage. By IHC, all five cases were positive for CK7 and p40; four were positive for GATA3; two were positive for CK20; and the mean Ki-67 proliferation index was 18%. TERT promoter mutation status were successfully performed in three cases, with one showing mutation (C228T) and two were wild type. All patients received postoperative chemotherapy. At a follow-up of 2 to 11 months, one patient died of unrelated causes, two patients developed metastases, and two were alive with no evidence of disease.Conclusions:LNUC is a histologic subtype of urothelial carcinoma with deceptively benign features but aggressive behavior, and appreciation of its unique infiltration patterns can aid in diagnosis and differential diagnosis. LNUC tends to coexist with small nested variant of urothelial carcinoma, suggesting these may represent different manifestations of the same urothelial carcinoma subtype.

Objective:To investigate the association of single nucleotide polymorphism at the estrogen receptor 1(ESR1) gene rs1801132 with the risk of brick-tea type skeletal fluorosis.Methods:The typical brick-tea type fluorosis areas in Qinghai, Xinjiang, and Inner Mongolia were selected as the survey sites for a cross-sectional study. An epidemiological questionnaire was conducted by the staffs on the sites for participants older than 16 years, and physical examination and X-ray diagnosis were performed. Brick tea, blood, and urine samples were collected at the same time. The diagnosis of skeletal fluorosis through X-ray was based on the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS/T 192-2008); The determination of tea's fluoride and urinary fluoride was performed by fluoride ion-selective electrode method; gene sequencing analysis of rs1801132 locus of ESR1 gene was done by Sequenom MassARRAY flight mass spectrometry system.Results:A total of 994 patients were included in this study. The total prevalence of skeletal fluorosis was 23.9% (238/994). The prevalence of skeletal fluorosis in Tibetans(39.9%, 123/308) was higher than those of Mongolian and Han nationality [22.2% (58/261), 13.4% (57/425), χ 2=20.435, 67.811, P < 0.05]. Based on binary logistic analysis, the daily tea fluoride intake ≤ 3.5 mg, urinary fluoride content ≤1.6 mg/L, and age ≤45 years were used as the reference groups, and then, when the daily tea fluoride intake > 7.0 mg ( OR=2.865, 95% CI: 1.923-4.268), urinary fluoride content > 1.6-3.2 mg/L ( OR=2.368, 95% CI: 1.686-3.326) and > 3.2 mg/L ( OR=3.559, 95% CI: 2.401-5.276), the age > 45-65 years old ( OR=2.361, 95% CI: 1.603-3.477) and > 65 years old ( OR=4.556, 95% CI: 2.845-7.296), the risk of fluorosis was higher than that of the reference group, respectively. When the daily tea fluoride intake was > 3.5-7.0 mg and the level of urinary fluoride was > 1.6-3.2 mg/L, G allele had a protective effect on skeletal fluorosis in Mongolian population (adjusted OR=0.207, 95% CI: 0.044-0.974); when the daily tea fluoride intake was > 3.5-7.0 mg, gender was male group, G allele had a protective effect on skeletal fluorosis in Han population (adjusted OR=0.315, 95% CI: 0.112-0.887). Conclusion:The single nucleotide polymorphism of the rs1801132 locus at the ESR1 gene may be associated with the risk of susceptibility to brick-tea type skeletal fluorosis in Mongolian and Han nationality.