OBJECTIVE To prepare hyaluronic acid （HA）-modified emodin （EMD）-contained multi-walled carbon nanotubes （MWCNTs） drug delivery system （HA-MWCNTs-EMD） and explore its in vitro inhibitory effect on breast cancer cells. METHODS EMD was loaded onto MWCNTs to prepare a drug delivery system MWCNTs-EMD； subsequently， the system was further modified with HA to obtain the drug delivery system HA-MWCNTs-EMD. The two drug delivery systems mentioned above were characterized. With free EMD as the reference， the drug release in vitro of the above two drug delivery systems was investigated； the uptake of EMD by two breast cancer cells （MCF-7， MDA-MB-231 cells） was detected. The impacts of the above two drug delivery systems on the expression of surface glycoprotein differentiation group 44 （CD44）， activity， apoptosis and lactate dehydrogenase （LDH） release of two breast cancer cells were detected. RESULTS The encapsulation efficiencies of MWCNTs-EMD and HA-MWCNTs-EMD were both （63.52±2.74）%， with drug loading rates of （25.01±1.83）% and （12.13± 1.96）%， particle sizes of （865.95±2.16） and （351.86±1.68） nm， polydispersity indexes of 0.54±0.02 and 0.23±0.01， and Zeta potentials of （23.87±0.14） and （－42.79±0.39） mV， respectively. The 2， 4， 6， 8， 10， 12 and 24-hour cumulative release rates of EMD in MWCNTs-EMD and HA-MWCNTs-EMD were significantly lower than those in free EMD， while the cumulative release rate of HA-MWCNTs-EMD was significantly higher than that of MWCNTs-EMD （P＜0.05）； the EMD uptakes of MWCNTs-EMD and HA-MWCNTs-EMD by the two types of breast cancer cells were significantly higher than their uptake of free EMD （P＜0.05）. Compared with the free EMD group， the MWCNTs-EMD and MWCNTs-EMD groups showed significantly higher apoptosis rate and LDH release， significantly lower surface CD44 expression （except for the MWCNTs-EMD group） and cell viability in both cell types， and the effect of HA-MWCNTs-EMD was more pronounced （P＜0.05）. CONCLUSIONS A novel drug delivery system HA-MWCNTs- EMD loaded with EMD is developed successfully； the drug delivery system has a certain slow-release effect， which can significantly reduce the activity of breast cancer cells， promote their apoptosis and increase the release of LDH， and the above anti- breast cancer effect is significantly stronger than that of free EMD and MWCNTs-EMD.

Objectives:To explore the relationship between insomnia symptoms and neuroticism in patients with COVID-19,and to explore the role of anxiety and psychological capital in the relationship.Methods:Totally 687 patients with COVID-19 were recruited from Shanghai Fangcang Hospital.The Athens Insomnia Scale(AIS),Eysenck Personality Questionnaire-Revised Short Scale for Chinese Neuroticism Subscale(EPQ-RSC-N),Self-Rat-ing Anxiety Scale(SAS)and Psychological Capital Questionnaire(PCQ)were used to measure insomnia symp-toms,neuroticism personality trait,anxiety symptoms and psychological capital levels.The deviation-corrected per-centile Bootstrap method was used to test the mediating effect,and the PROCESS program was used to test the moderated effect.Results:The detection rate of insomnia symptoms was 49.93％.The AIS scores were lower in male patients than in female patients(P＜0.01).The SAS scores partly mediated the relationship between neuroti-cism scores and AIS scores,with an effect size of 0.03,accounting for 18.29％of the total effect.With the im-provement of PCQ scores,the predictive effect of SAS scores on AIS scores gradually decreased(β=-0.01,t=-4.41,P＜0.001).Conclusions:Anxiety symptoms in patients with COVID-19 play a partial mediating role in the positive relationship between insomnia symptoms and neuroticism.The psychological capital moderates the relation-ship between insomnia and anxiety symptoms.

Epigenomic imbalance drives abnormal transcriptional processes,promoting the onset and progression of cancer.Although defective gene regulation generally affects carcinogenesis and tumor suppression networks,tumor immunogenicity and immune cells involved in antitumor responses may also be affected by epigenomic changes,which may have significant implications for the development and application of epigenetic therapy,cancer immunotherapy,and their combinations.Herein,we focus on the impact of epigenetic regulation on tumor immune cell function and the role of key abnormal epigenetic processes,DNA methylation,histone post-translational modification,and chromatin structure in tumor immunogenicity,and introduce these epigenetic research methods.We emphasize the value of small-molecule inhibitors of epigenetic modulators in enhancing antitumor immune responses and discuss the challenges of developing treatment plans that combine epigenetic therapy and immuno-therapy through the complex interaction between cancer epigenetics and cancer immunology.

Objective:To design a multi-epitope antigen of norovirus(NoV)based on bioinformatics technology and to pre-pare and characterize it.Methods:Bioinformatics methods were used to construct and analyze the NoV multi-epitope antigen NoV-ZH.Recombinant proteins were prepared and characterized by prokaryotic expression system,and monoclonal antibodies were prepared by animal immunization and hybridoma technology,and initially applied in colloidal gold platform.Results:The designed multi-epitope antigen had a large proportion of random curls in the secondary structure,with theoretical molecular mass and isoelectric point(PI)of 13.1 ku and 7.16,which were stable and hydrophilic.It had good immunogenicity and could activate humoral and cellular immune re-sponses.The proteins prepared by ligating pET-28a(+)and pET-32a vectors with antigenic sequences were expressed as inclusion body proteins and soluble proteins,respectively.A pair of paired antibodies was obtained by animal immunization and hybridoma tech-nique,and applied to colloidal gold test strips with a sensitivity of 0.5 ng/ml,and the test strips could specifically bind two genotypes of NoV recombinant capsid proteins.Conclusion:The successful preparation and characterization of multi-epitope antigen of norovirus provides a reference for the subsequent exploration of NoV universal detection targets and the development of diagnostic raw materials.

【Objective】 To establish a routine screening method for unexpected antibodies of blood donors, analyze the results of centralized screening for unexpected antibody of blood donors in the blood center, and compare the cost of centralized and decentralized screening modes. 【Methods】 A total of 35 591 blood donors were screened for unexpected antibodies from March 31, 2021 to July 31, 2021, using microcolumn gel method. Unexpected antibody screening reactive samples were further confirmed by the Transfusion Research Institute of Shenzhen Blood Center, and the demographic characteristics were further determined through the analysis of unexpected antibody positive population. The direct cost and indirect cost of centralized and decentralized unexpected antibody screening mode were compared. 【Results】 Forty unexpected antibody positive samples were confirmed in Shenzhen, with the positive rate at 0.11%(40/35 591), among which MNS, Rh and Lewis system accounted for 35% (14/40), 32.5% (13/40) and 17.5% (7/40), respectively. Males and females accounted for 45% (18/40) and 55% (22/40), respectively (P<0.01). No significant difference was noticed by age and repeated-donor or not (P>0.05). Unexpected antibody screening in a centralized way saved about 1.16 million yuan per year. 【Conclusion】 It is necessary to carry out unexpected antibody screening for all blood donors, and centralized screening is more economical than decentralized screening.

Objective:To analyze the surgical methods of operable breast cancer and analyze the follow-up results.Methods:A retrospective analysis of the clinical and pathological data of 636 operable breast cancer patients admitted to Zibo First Hospital from July 2008 to April 2018, including the clinical stage, pathological staging. Analyze of the proportion of four surgical methods, and through follow-up, analyze the treatment effect of different surgical methods.Results:All patients are female, aged 26-80 years, the clinical stage of 636 patients: Tis 18 cases, stage Ⅰ 143 cases, stage Ⅱ 354 cases, stage Ⅲ 114 cases, stage Ⅳ 7 cases. There are four types of surgery: ① breast conserving surgery + sentinel lymph node biopsy in 124 cases (19.50%); ② breast conserving surgery + axillary lymph node dissection in 39 cases (6.13%); ③ mastectomy + sentinel lymph node biopsy in 163 cases (25.63%); ④ modified radical surgery in 310 cases (48.74%). Sentinel lymph node biopsy in 427 cases (67.14%), success in 404 patients (94.61%); all patients with lymph node negative 384 cases (60.38%). Follow-up for 1 to 9 years, 11 cases of local recurrence after breast-conserving surgery, It accounted for 6.75% of breast-conserving surgery; 43 cases of local recurrence of chest wall after mastectomy, accounting for mastectomy 9.09%; 33 cases of recurrence and metastasis of axillary lymph nodes and supraclavicular lymph nodes, 4 cases of axillary recurrence after sentinel lymph node biopsy.Conclusions:The proportion of breast-conserving surgery in this group of patients was high and the local recurrence rate of breast-conserving surgery was less than that of mastectomy group; the proportion of simple modified radical surgery declined further; patients with axillary lymph node metastasis were less in the whole group. The choice of reasonable operation method is an important factor to improve the prognosis of breast cancer.

Objective:To construct a prognosis associated micro RNA(miRNA) prediction model based on bioinformatics analysis and evaluate its application value in pancreatic cancer patients.Methods:The retrospective cohort study was conducted. The clinicopathological data of 171 pancreatic cancer patients from the Cancer Genome Atlas (TCGA) (https: //cancergenome.nih.gov/) between establishment of database and September 2017 were collected. There were 93 males and 78 females, aged from 35 to 88 years, with a median age of 65 years. Of the 171 patients, 64 had complete clinicopathological data. Patients were allocated into training dataset consisting of 123 patients and validation dataset consisting of 48 patients using the random sampling method, with a ratio of 7∶3. The training dataset was used to construct a prediction model, and the validation dataset was used to evaluate performance of the prediction model. Nine pairs of miRNA sequencing data (GSE41372) of pancreatic cancer and adjacent tissues were downloaded from Gene Expression Omnibus database. The candidate miRNAs were selected from differentially expressed miRNAs in pancreatic cancer and adjacent tissues for LASSO-COX regression analysis based on the patients of training dataset. A prognosis associated miRNA prediction model was constructed upon survival associated miRNAs which were selected from candidate differentially expressed miRNAs. The performance of prognosis associated miRNA prediction model was validated in training dataset and validation dataset, the accuracy of model was evaluated using the area under curve (AUC) of the receiver operating characteristic curves and the efficiency was evaluated using the consistency index (C-index). Observation indicarors: (1) survival of patients; (2) screening results of differentially expressed miRNAs; (3) construction of prognosis associated miRNA model; (4) validation of prognosis associated miRNA model; (5) comparison of clinicopathological factors in pancreatic cancer patients; (6) analysis of factors for prognosis of pancreatic cancer patients; (7) comparison of prediction performance between prognosis associated miRNA model and the eighth edition TNM staging. Measurement data with normal distribution were represented as Mean± SD, comparison between groups was analyzed by the student- t test, and comparison between multiple groups was analyzed by the AVONA. Measurement data with skewed data were represented as M (range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Ordinal data were analyzed using the rank sum test. Correlation analysis was conducted based on count data to mine the correlation between prognosis associated miRNA model and clinicopathological factors. COX univariate analysis and multivariate analysis were applied to evaluate correlation with the results described as hazard ratio ( HR) and 95% confidence interval ( CI). HR<1 indicated the factor as a protective factor, HR>1 indicated the factor as a risk factor, and HR equal to 1 indicated no influence on survival. The Kaplan-Meier method was used to draw survival curve and calculate survival rates, and the Log-rank test was used for survival analysis. Results:(1) Survival of patients: 123 patients in the training dataset were followed up for 31-2 141 days, with a median follow-up time of 449 days. The 3- and 5-year survival rates were 16.67% and 8.06%. Forty-eight patients in the validation dataset were followed up for 41-2 182 days, with a median follow-up time of 457 days. The 3- and 5-year survival rates were 15.63% and 9.68%. There was no significant difference in the 3- or 5-year survival rates between the two groups ( χ2=0.017, 0.068, P>0.05). (2) Screening results of differentially expressed miRNAs. Results of bioinformatics analysis showed that 102 candidate differentially expressed miRNAs were selected, of which 63 were up-regulated in tumor tissues while 39 were down-regulated. (3) Construction of prognosis associated miRNA model: of the 102 candidate differentially expressed miRNAs, 5 survival associated miRNAs were selected, including miR-21, miR-125a-5p, miR-744, miR-374b, miR-664. The differential expression patterns of pancreatic cancer to adjacent tissues were up-regulation, up-regulation, down-regulation, up-regulation, and down-regulation, respectively, with the fold change of 4.00, 3.43, 3.85, 2.62, and 2.35. A prognostic expression equation constructed based on 5 survival associated miRNAs = 0.454×miR-21 expression level-0.492×miR-125a-5p expression level-0.49×miR-744 expression level-0.419×miR-374b expression level-0.036×miR-664 expression level. (4) Validation of prognosis associated miRNA model: The C-index of prognosis associated miRNA model was 0.643 and 0.642 for the training dataset and validation dataset, respectively. (5) Comparison of clinicopathological factors in pancreatic cancer patients: results of COX analysis showed that the prognosis associated miRNA model was highly related with pathological T stage and location of pancreatic cancer ( Z=45.481, χ2=10.176, P<0.05). (6) Analysis of factors for prognosis of pancreatic cancer patients: results of univariate analysis showed that pathological N stage, radiotherapy, molecular targeted therapy, score of prognosis associated miRNA model were related factors for prognosis pf pancreatic cancer patients ( HR=2.471, 0.290, 0.172, 2.001, 95% CI: 1.012-6.032, 0.101-0.833, 0.082-0.364, 1.371-2.922, P<0.05). Results of multivariate analysis showed that molecular targeted therapy was an independent protective factor for prognosis of pancreatic cancer patients ( HR=0.261, 95% CI: 0.116-0.588, P<0.05) and score of prognosis associated miRNA model≥1.16 was an independent risk factor for prognosis of pancreatic cancer patients ( HR=1.608, 95% CI: 1.091-2.369, P<0.05). (7) Comparison of prediction performance between prognosis associated miRNA model and the eighth edition TNM staging: in the training dataset, there was a significant difference in the prediction probability for 3- and 5-year survival of pancreatic cancer patients between prognosis associated miRNA model and the eighth edition TNM staging ( Z=-1.671, -1.867, P<0.05). The AUC of the prognosis associated miRNA model and the eight edition TNM staging for 3- and 5-year survival prediction was 0.797, 0.935 and 0.737 , 0.703, with the 95% CI of 0.622-0.972, 0.828-1.042 and 0.571-0.904 , 0.456-0.951. The C-index was 0.643 and 0.534. In the validation dataset, there was a significant difference in the prediction probability for 3- and 5-year survival of pancreatic cancer patients between prognosis associated miRNA model and the eighth edition TNM staging ( Z=-1.729, -1.923, P<0.05). The AUC of the prognosis associated miRNA model and the eight edition TNM staging was 0.750, 0.873 and 0.721 , 0.703, with the 95% CI of 0.553-0.948, 0.720-1.025 and 0.553-0.889, 0.456-0.950, respectively. The C-index was 0.642 and 0.544. Conclusions:A prognosis associated miRNA prediction model can be constructed based on 5 survival associated miRNAs in pancreatic cancer patients, as a complementation to current TNM staging and other clinicopathological parameters, which provides individual and accurate prediction of survival for reference in the clinical treatment.

Objective:To observe the clinical effect of the improved eyebrow lifting operation through the incision under the eyebrow to improve the upper eyelid skin relaxation and explore its application scope.Methods:From March to October 2019, 32 female patients in the outpatient department of Huangsi Plastic Surgery Hospital underwent the improved eyebrow lifting operation through the incision under the eyebrow.Results:After the improved eyebrow lifting procedure, the blepharoptosis of the upper eyelid in 32 patients was significantly improved, and there was no operative complication, and the eyebrow shape and eyebrow arch fullness were satisfactory.Conclusions:The eyebrow arch is full, and the eyebrow shape and the distance between eyebrow and eye are not changed significantly after the operation.

Objective:To investigate the risk factors of lymph node metastasis for patients with colorectal cancer in T 3 and T 4, and to provide a reference for clinical diagnosis and treatment. Methods:The clinicopathological data of 1 112 patients with colorectal cancer in T 3 and T 4 who underwent radical resection of colorectal cancer in Xijing Digestive Disease Hospital from January 2008 to December 2017 were retrospectively analyzed. The correlation between lymph node metastasis status and the clinicopathological factors as well as tumor markers was analyzed. The related risk factors of lymph node metastasis were analyzed by using logistic multivariate regression analysis. Results:Univariate analysis showed that there was no statistically significant difference in the incidence of lymph node metastasis among colorectal cancer patients stratified by gender, age and tumor location (all P > 0.05). The different tumor diameter [<5 cm and ≥5 cm: 37.75% (211/559), 52.26% (289/553), χ2 = 23.666, P < 0.01], general type [infiltration, ulcer, parasol, bulge: 37.04% (20/54), 47.52% (432/909), 34.33% (23/67), 69.51% (57/82), χ2 = 13.787, P = 0.003], degree of differentiation [highly-differentiated, moderately-differentiated, poorly-differentiated: 34.11% (102/299), 49.00% (317/647), 48.80% (81/166), χ2 = 19.771, P < 0.01], mismatch repair deficiency (dMMR) [yes and no: 26.34% (64/243), 50.17% (436/869), χ2 = 43.996, P < 0.01], neurological invasion [yes and no: 48.17% (421/874), 33.20% (79/238), χ2 = 16.954, P < 0.01], vascular invasion [yes and no: 79.16% (338/427), 23.65% (162/685), χ2 = 327.493, P < 0.01] and preoperative carcino-embryonic antigen (CEA) [positive (≥5 mg/ml) and negative (<5 mg/ml): 52.87% (249/471), 39.16% (251/641), χ2 = 20.162, P < 0.01] and CA199 [positive (≥35 U/ml) and negative (<35 U/ml): 59.33% (124/209), 41.64% (376/903), χ2 = 21.465, P < 0.01] had statistically significant differences in the incidence of lymph node metastasis for above stratified patients. Logistic multivariate regression analysis showed that vascular invasion and preoperative CA199-positive were independent risk factors for lymph node metastasis in patients with colorectal cancer in T 3 and T 4 ( OR = 13.006, 95% CI 9.329-17.276, P < 0.01; OR = 2.194, 95% CI 1.513-3.181, P < 0.01), and dMMR-positive was a protective factor for lymph node metastasis ( OR = 0.279, 95% CI 0.190-0.411, P < 0.01). Conclusions:Vascular invasion is the main risk affecting factor for the lymph node metastasis of patients with colorectal cancer in T 3 and T 4. The detection of preoperative tumor marker CA199 can be used as an index to predict the lymph node metastasis of patients with colorectal cancer in T 3 and T 4. To a certain extent, it can provide a reference for the diagnosis and treatment of patients with colorectal cancer in T 3 and T 4.

Objective: To describe the regional and population-related differences in skeletal muscle mass and handgrip strength across the 10 regions of China. Methods: 24 533 participants aged 38-88 years from the second resurvey of China Kadoorie Biobank were included in our analyses. Appendicular and trunk skeletal muscle mass were assessed using the bioelectrical impedance analysis (TANITA). Handgrip strength was measured using Jamar hand-held dynamometer. Low muscle mass and low muscle strength were defined as the lowest quintile of height-adjusted appendicular muscle mass or handgrip strength according to the Consensus Report of the Asian Working Group for Sarcopenia. We analyzed the mean value of absolute muscle mass, height-adjusted muscle mass, weight-adjusted muscle mass and handgrip strength. We also reported the prevalence of low muscle mass and low muscle strength. Results: The average appendicular and total skeletal muscle mass were (22.0±0.02) kg and (49.7±0.05) kg in men, which were higher than in women [(15.9±0.02) kg and (37.2±0.04) kg, respectively]. The handgrip strength was (32.6±0.06) kg in men, which was higher than (19.9±0.05) kg in women. The absolute muscle mass was higher in north area and urban region (P<0.001). The weight-adjusted muscle mass showed reverse patterns of regional difference compared with height-adjusted muscle mass. Both muscle mass and handgrip strength decreased by age (trend P<0.001), with a larger decline observed in handgrip strength. According to AWGS criteria, the proportions of low muscle mass and strength increased by age. Among participants over 80 years old, the prevalence of low muscle mass and strength were 56.2% and 74.5% in men, and 35.7% and 66.0% in women. Conclusions Levels of skeletal muscle mass and strength varied greatly among people from 10 regions and among participants with different demographic characteristics. The prevalence of low muscle mass and strength was extremely high in elderly.