Objective:To investigate the expressions of cystathionine-β-synthase(CBS)and cystathionine-γ-lyase(CSE)and their effects on the malignant biological behaviours of breast cancer cells,and to elucidate their mechanisms.Methods:The breast cancer tissue and paracancerous normal tissue from 15 cases of patients were selected,and RT-qPCR and Western blotting methods were used to detect the mRNA and protein expression levels of CBS and CSE in breast cancer tissue,paracancerous normal tissue,MCF-7 cells,and MDA-MB-231 cells.The MCF-7 cells were divided into siNC group(transfected with siNC)and siCBS group(transfected with siCBS),and the MDA-MB-231 cells were divided into ovNC group(transfected with CSE over-expression empty plasmid)and ovCSE group(transfected with CSE over-expression plasmid).CCK8 assay was used to detect the proliferation activities of breast cancer cells in various groups,Transwell assay was used to detect the numbers of migration and invasion cells in various groups,and Western blotting method was used to detect the protein expression levels of E-cadherin,N-cadherin and Vimentin proteins in the breast cancer cells in various groups.Results:Compared with paracancerous normal tissue,the expression levels of CBS and CSE mRNA and proteins in breast cancer tissue were increased(P＜0.05 or P＜0.01).Compared with MDA-MB-231 cells,the CBS mRNA expression level in the MCF-7 cells was increased(P＜0.05);compared with MCF-7 cells,the expression level of CSE protein in the MDA-MB-231 cells was decreased(P＜0.05).Compared with siNC group,the proliferation activity,the numbers of migration and invasion cells,the expression levels of N-cadherin and Vimentin proteins in the MCF-7 cells in siCBS group were significantly decreased(P＜0.05),and the expression level of E-cadherin protein was increased(P＜0.05).Compared with ovNC group,the proliferation activity,the numbers of migratoin and invasion cells,and the expression levels of N-cadherin and Vimentin proteins in the MDA-MB-231 cells in ovCSE group were increased(P＜0.05),while the expression level of E-cadherin protein was significantly decreased(P＜0.05).Conclusion:The expressions of CBS and CSE are upregulated in breast cancer tissue,and high levels of CBS and CSE promote proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of breast cancer cells.

Objective:To explore the standardized performance of a FISH probe before clinical detection.Methods:The probe sensitivity and specificity of ETV6/RUNX1 were analyzed via interphase and metaphase FISH in 20 discarded healthy bone marrow samples. The threshold system of the probe was established using an inverse beta distribution, and an interpretation standard was established. Finally, a parallel-controlled polymerase chain reaction detection study was conducted on 286 bone marrow samples from patients at our hospital. The clinical sensitivity, specificity, and diagnostic coincidence rate of ETV6/RUNX1 FISH detection were analyzed, and the diagnostic consistency of the two methods was analyzed by the kappa test.Results:The probe sensitivity and specificity of the ETV6/RUNX1 probe were 98.47% and 100%, respectively. When 50, 100, and 200 cells were counted, the typical positive signal pattern cutoffs were 5.81%, 2.95%, and 1.49%, respectively, and the atypical positive signal pattern cutoffs were 13.98%, 9.75%, and 6.26%, respectively. The clinical sensitivity of FISH was 96.1%, clinical specificity was 99.6%, diagnostic coincidence rate was 99.00%, diagnostic consistency test kappa value was 0.964, and P value was <0.001.Conclusion:For FISH probes without a national medical device registration certificate, standardized performance verification and methodology performance verification can be performed using laboratory developed test verification standards to ensure a reliable and accurate reference basis for clinical diagnosis and treatment.

Objective: To investigate the prevalence and relevant factors of eating out-of-home among middle school students in Wuhan, and to analyze its association with overweight/obesity, so as to provide scientific basis for the prevention of overweight/obesity among middle school students. Methods: From March to April 2023, 1 654 middle school students in Wuhan were selected by convenient sampling method for a questionnaire survey. The multivariate Logistic regression model was used to analyze the relevant factors of eating out-of-home and to explore its relationship with overweight/obesity among middle school students. Results: On weekdays, 57.19% of the middle school students ate out-of-home at least once a week. On weekends, 81.38% of the participants ate out-of-home at least once a week. The multivariate analysis showed non -resident students were more likely to eat out-of-home on weekdays (OR=4.32, 95%CI =2.10-8.90, P <0.05). Middle school students whose fathers with educational backgrounds of high school, technical secondary school or college above, and families with per capita monthly income levels of 2 000-5 000 and >5 000 yuan, and average monthly food expenses of 500-<1 000 and 1 000-<1 500 yuan were more likely to eat out-of-home on weekends ( OR= 2.12 , 2.77; 2.58, 3.32; 1.70, 1.98, P <0.05). Middle school students with qualified nutrition and health knowledge scores were more likely to eat out-of-home on weekends than those with excellent nutrition and health knowledge scores ( OR=1.51, 95%CI = 1.12 -2.06, P <0.05). Students who ate out-of-home once or twice per week on weekends had 1.50 times higher risk of overweight/obesity than those who ate out-of-home none per week on weekends (adjusted OR =1.50, 95% CI =1.05-2.15, P <0.05). Conclusion Eating out-of-home is common among middle school students in Wuhan. Middle school students should actively cultivate awareness of healthy eating, and decrease the frequency of eating out-of-home to reduce the incidence of overweight and obesity .

Background::Heterogeneity of tumor cells and the tumor microenvironment (TME) is significantly associated with clinical outcomes and treatment responses in patients with urothelial carcinoma (UC). Comprehensive profiling of the cellular diversity and interactions between malignant cells and TME may clarify the mechanisms underlying UC progression and guide the development of novel therapies. This study aimed to extend our understanding of intra-tumoral heterogeneity and the immunosuppressive TME in UC and provide basic support for the development of novel UC therapies.Methods::Seven patients with UC were included who underwent curative surgery at our hospital between July 2020 and October 2020. We performed single-cell RNA sequencing (scRNA-seq) analysis in seven tumors with six matched adjacent normal tissues and integrated the results with two public scRNA-seq datasets. The functional properties and intercellular interactions between single cells were characterized, and the results were validated using multiplex immunofluorescence staining, flow cytometry, and bulk transcriptomic datasets. All statistical analyses were performed using the R package with two-sided tests. Wilcoxon-rank test, log-rank test, one-way analysis of variance test, and Pearson correlation analysis were used properly.Results::Unsupervised t-distributed stochastic neighbor embedding clustering analysis identified ten main cellular subclusters in urothelial tissues. Of them, seven urothelial subtypes were noted, and malignant urothelial cells were characterized with enhanced cellular proliferation and reduced immunogenicity. CD8 + T cell subclusters exhibited enhanced cellular cytotoxicity activities along with increased exhaustion signature in UC tissues, and the recruitment of CD4 + T regulatory cells was also increased in tumor tissues. Regarding myeloid cells, coordinated reprogramming of infiltrated neutrophils, M2-type polarized macrophages, and LAMP3 + dendritic cells contribute to immunosuppressive TME in UC tissues. Tumor tissues demonstrated enhanced angiogenesis mediated by KDR + endothelial cells and RGS5 +/ACTA2 + pericytes. Through deconvolution analysis, we identified multiple cellular subtypes may influence the programmed death-ligand 1 (PD-L1) immunotherapy response in patients with UC. Conclusion::Our scRNA-seq analysis clarified intra-tumoral heterogeneity and delineated the pro-tumoral and immunosuppressive microenvironment in UC tissues, which may provide novel therapeutic targets.

ObjectiveTo investigate effect of conducting training of autism spectrum disorder （ASD） early screening skill on improving the ability to early identify ASD of medical staffs in primary care hospitals. MethodsIn September 2021， the training of ASD early screening skills was carried out for medical staffs from 20 primary care hospitals in Chengdu. After training， the training effect was evaluated. The numbers of referrals from primary care hospitals to superior hospitals， confirmed ASD as well as their average diagnostic age of children with ASD before and after training were used as evaluation indicators. ResultsAfter training， the number of children with suspected ASD referred by primary care hospitals was more than that before training ［（16.65±11.60） vs. （3.40±2.23）， t=5.431， P<0.01］， the number of children diagnosed with ASD was more than that before training［（6.85±4.93） vs. （2.45±1.67）， t=4.171， P<0.01］， and the differences were statistically significant. As for the diagnosed age of ASD children， after training， the average age was lower than that before training ［（34.95±11.67） vs. （42.2±14.64）， t=-2.553， P=0.019］. ConclusionTraining of ASD early screening skills for medical staffs in primary care hospitals may help to improve their ability to early screening ASD children.

Objective:To investigate the expression of circ0008234 and miR-1205 in breast cancer and the oncology characteristics of CT.Methods:40 patients with breast cancer were collected as observational objects from Jan.2018 to Dec.2018 and 40 patients without breast cancer were selected as the control group. The expression levels of circ0008234 and miR-1205 of all patients were estimated by quantitative real-time polymerase chain reaction. Bioinformatics was used for the predictions of circ0008234 target miRNA. A dual-luciferase reporter assay was used to confirm the interaction between circ0008234 and miR-1205. All patients of the observation group were examined by multi-slice CT. CT images were analyzed through observing tumor size, shape, calcification area, lymph node metastasis and margin. The correlation between CT signs and the expression of circ0008234 and miR-1205 was further analyzed.Results:The levels of circ0008234 of observation group were significantly higher than those of the control group (2.23±0.86, 1.07±0.37, P=0.00) , but the expression levels of miR-1205 were lower than the control group (0.76±0.29, 1.04±0.29, P=0.00) . In the observation group, there were no significant differences in the expression of circ0008234 and miR-1205 among patients with different tumor marginal morphology and micro calcifications. However the expression of circ0008234 in patients with regular lump form were significantly higher than those in patients with inregular form (2.52±0.88, 1.91±0.74, P=0.025) , and the expression of miR-1205 were lower than those in patients with inregular form (0.66±0.30, 0.86±0.25, P=0.025) . In the observation group, the expression of circ0008234 in patients with mass diameter≥2 cm were significantly higher than those in patients with mass diameter<2 cm (2.59±0.95, 1.69±0.17, P=0.001) , but the expression of miR-1205 were lower than those in patients with mass diameter<2 cm (0.65±0.21, 0.92±0.33, P=0.003) . In the observation group, the expression of circ0008234 in patients with lymph node metastasis were higher than those without lymph node metastasis (2.55±1.09, 1.94±0.44, P=0.022) , but the expression of miR-1205 in patients with lymph node metastasis were lower than those without lymph node metastasis (0.65±0.26, 0.85±0.30, P=0.027) . miR-1205 was verified as a direct target of circ0008234 by luciferase assay. circ0008234 could negatively regulate the expression of miR-1205. Conclusion:There is a correlation between CT imaging signs and the expression of circ0008234 and miR-1205 in patients with breast cancer, which can provide more reference for the judgment of malignant degree and prognosis of patients with breast cancer.

Objective: To explore the duration of intestinal detoxification and its related factors infected with Norovirus, and to provide scientific evidences for the prevention and control of acute gastroenteritis caused by Norovirus in the future. Methods: Follow up and sampling program on 115 employees in a restaurant outbreak of Norovirus in Chengdu in 2017 was carried out. Anal swab was collected every 3 d or 7 d for nucleic acid detection, and the experimental results were statistically analyzed. Results: The positive rates of Norovirus in restaurant employee were 14.78%, 7.89%, 3.51% and 0.88% on the 7^th, 11^th, 17^th and 24^th day, respectively. During the follow-up period, the positive rates of Norovirus decreased as time increased. The positive rate of all employees was up to 14.78% on the 7^th day. The positive rates had no significant difference between different sex and between different types of work. The duration of detoxification averaged 14.94 days and the maximum of detoxification time was on 28 days. The detoxification period of treatment case was shorter than that of asymptomatic carriers. Conclusions There were asymptomatic carriers in restaurant employees during the outbreak of Norovirus that their role as a source of infection in the transmission of Norovirus should not be neglected. In the treatment of the epidemic, we should strengthen the sampling and testing of practitioners, and strictly implement isolation measures for positive detection.

Objective To explore the four tumor markers of alpha-fetoprotein ( AFP ), α-L-fucosidase( AFU), carbohydrate antigen 199 ( CA199) and carcinoembryonic antigen ( CEA) and their combined use for primary hepatocellular carcinoma (HCC) diagnosis and treatment value. Methods From February 2016 to August 2018,92 patients with primary hepatocellular carcinoma (HCC group),79 patients with benign liver disease (chronic hepatitis and cirrhosis group) and 99 healthy adults (control group) were selected as subjects. The serum levels of four tumor markers in different populations were compared. Results The serum levels of four tumor markers ( AFP ( 192. 4 ± 89. 3) μg／L、AFU( 78. 6 ± 25. 8) U／L、CA199 (107. 2 ± 59. 5) U／mL、 CEA ( 37. 9 ± 14. 9) μg／L) were significantly higher than those of benign liver disease group(AFP( 17. 4 ± 6. 3) μg／L、AFU( 35. 4 ± 17. 2) U／L、CA199( 29. 3± 15. 2) U／mL、CEA( 4. 9 ±1. 7) μg／L) and normal people( AFP(4. 8±1. 1) μg／L、AFU(12. 2±3. 6) U／L、CA199( 6. 4± 2. 3) U／mL、CEA(1. 8±0. 4) μg／L) . There differences had significant ( all P＜0. 05) . The abnormal rate of single factor in hepatocellular carcinoma group ( AFP 84. 8%, AFU 52. 2%, CA199 41. 3%, CEA35. 9%) was significantly higher than that in benign liver disease group ( AFP 15. 2%, AFU 19. 0%, CA19916. 5%, CEA13. 9%) and normal group (AFP 4. 0%,AFU 5. 0%,CA199 3. 0%,CEA 6. 0% ug／L),the difference was statistically significant ( all P＜0. 05) . The highest sensitivity was AFP ( 84. 8%) and the highest specificity was AFP and CA199 (91. 0%). The sensitivity of combined detection was 94. 6% higher than that of single index ( AFP 84. 8%, AFU52. 2%, CA199 41. 3%, CEA35. 9%) . Conclusion The combined detection of AFP,AFU,CA199 and CEA can increase the sensitivity of diagnosis of hepatocellular carcinoma and reduce the rate of missed diagnosis, which will be beneficial to the early diagnosis and treatment of hepatocellular carcinoma.

Objective To investigate the clinical value of joint detection of six tumor markers in patients with colorectal cancer. Methods Eighty?six patients with colorectal cancer were included in the study group,86 healthy subjects were selected as the control group at the same period. The difference of tumor markers in different groups,tumor stages and prognosis were compared. Results The levels of carcinoembryonic antigen (CEA),carbohydrate antigen 19?9 (CA19?9),carbohydrate antigen 242 (CA242),carbohydrate antigen 72?4 ( CA72?4) , carbohydrate antigen 125 ( CA125 ) and carbohydrate antigen 50 ( CA50 ) in study group were significantly higher than those in the control group (CEA: (22. 5±6. 2)μg/L vs. (2. 2±1. 0)μg/L;CA19?9:(95. 7±27. 3) U/ml vs. (17. 1±9. 5) U/ml;CA242:(29. 5±8. 3) U/ml vs. (6. 0±2. 7) U/ml;CA72?4:(21. 6 ±5. 1) U/ml vs. (3. 6±1. 2) U/ml;CA125:(95. 4±32. 8) U/ml vs. (18. 9±8. 4) U/ml;CA50:(51. 8±20. 6)μg/L vs. (8. 3±3. 7)μg/L,t=29. 98,25. 22,24. 97,31. 86,20. 95,19. 27,P<0. 05). Among the single index detections,the sensitivity and negative predictive value of CA72?4 were the highest ( 61. 6%, 68. 3%) , the specificity of CA19?9 was the highest( 91. 9%) ,the positive predictive value of CEA was the highest ( 80. 4%) . The sensitivity,positive predictive value and negative predictive value of the joint detection were all higher than those in each single index detection (80. 3%,87. 3%,74. 1%). The levels of CEA,CA19?9,CA242,CA72?4, CA125 and CA50 in patients with stage III and IV were significantly higher than those in patients with stageⅠandⅡ(CEA:(32. 7±7. 1)μg/L vs. (15. 9±4. 4)μg/L;CA19?9:(127. 8±33. 7) U/ml vs. (52. 5±13. 8) U/ml;CA242:(40. 3±12. 7) U/ml vs. (23. 5±8. 6) U/ml;CA72?4:(37. 6±10. 2) U/ml vs. (13. 6±4. 1) U/ml;CA125:(128. 9±38. 4) U/ml vs. (59. 7±12. 8) U/ml;CA50:(88. 3±23. 7)μg/L vs. (41. 8±15. 6)μg/L,t=13. 04,13. 32,7. 11,14. 06,10. 99,10. 64,P<0. 05) . The levels of CEA,CA19?9,CA242,CA72?4,CA125 and CA50 in the recurrent metastasis group were significantly higher than those in the non?recurrent metastasis group ( CEA:( 37. 7 ± 8. 6 ) μg/L vs. ( 3. 8 ± 1. 7 ) μg/L;CA19?9:( 110. 5 ± 29. 4 ) U/ml vs. ( 25. 5 ± 13. 8 ) U/ml;CA242:( 33. 6 ± 10. 3 ) U/ml vs. ( 15. 5 ± 6. 6 ) U/ml;CA72?4:( 33. 1 ± 15. 3 ) U/ml vs. ( 9. 3 ± 3. 0 ) U/ml;CA125:(113. 4±31. 7) U/ml vs. (28. 7±7. 8) U/ml;CA50:(55. 4±14. 6)μg/L vs. (16. 8±9. 6)μg/L,t=29. 04,18. 31,9. 86,11. 47,19. 28,14. 65,P<0. 05) . Conclusion The joint detection of six markers can further improve the sensitivity, positive predictive value and negative predictive value of diagnosis, and can provide a more reliable basis for the auxiliary diagnosis of colorectal cancer.

Objective To investigate the effect of dexmedetomidine on the serum concentrations of S-100 β protein and neurone specific enolase (NSE) in elderly patients undergoing surgery.Methods One hundred ASA Ⅱ or Ⅲ patients,aged 65 ～75 yr with a body mass index of ＜25 kg/m2,scheduled for elective hip joint replacement surgery under general anesthesia,were randomly divided into 2 groups (n =50):control group (group C) and dexmedetomidine group (group D).Dexmedetomidine 0.5 μg/kg was infused intravenously over 15 min before anesthesia induction in group D and was maintained 0.4 μg/(kg·h) until 30 min before operation end,while the same volume of normal saline was infused in group C.Anesthesia was induced by iv injection of sufentanil 0.4 μg/kg and propofol 1 ～ 2 mg/kg until loss of consciousness.Tracheal intubation was facilitated with 0.6 mg/kg rocuronium bromide and the patients were mechanically ventilated.Anesthesia was maintained with sevoflurane by target controlled inhalation (TCI) (end-tidal concentration set at 1％～3％) and sufentanil 0.2 μg/(kg·h) in both groups.BIS was maintained at 40～60 during operation.Venous blood samples were obtained for determination of serum concentrations of S-100β protein and NSE before anesthesia (baseline),operation end and at 12h after operation.Patients were sent to Intensive Care Unit when operation end.All the patients were assessed for the development of delirium by experience research staff using Confusion Assessment Method for Intensive Care Unit.The incidence of postoperative delirium within 24h after operation were recorded,and compared between the two groups.Results The serum concentrations of S-100β protein and NSE was significantly increased at T1～2 than at T0 in two groups.The total dose of each anesthetic (propofol,sevoflurane),the serum concentrations of S-1005 protein and NSE,and the incidence of postoperative delirium were significantly lower in group D than in group C (P＜0.05).Conclusions Dexmedetomidine can reduce the serum concentrations of S-100 β protein and NSE,and can also reduce the occurrence of postoperative delirium in elderly patients undergoing hip joint replacement surgery.Dexmedetomidine can provides cerebral protection in elderly patients undergoing surgery.