Objective The purpose of the study is to breed homozygous interferon-γ knockout(IFN-γ-/-)mice and optimize the breeding strategies to achieve continuous and stable reproduction of IFN-γ-/-mice,which could be used as an ideal animal model for fundamental research.Methods Initially,heterozygous IFN-γ knockout(IFN-γ+/-)C57BL/6J mice were used as the parental generation for breeding.Subsequently,3 breeding strategies were employed using the offspring:(1)female heterozygotes mated with male heterozygotes;(2)male homozygotes mated with female heterozygotes;(3)female homozygotes mated with male homozygotes.The number and survival rate of IFN-γ-/-mice were compared across the three breeding strategies to determine the optimal breeding strategy.Under the optimal strategy,the effects of female mating age and diet type on the reproductive performance of IFN-γ-/-mice were further evaluated.Data from the first three litters of 60 IFN-γ-/-female mice,including litter size,number of weaning survivors,and weaning survival rate,were recorded and analyzed.In addition,the effects of dietary supplementation of pregnant mice and environmental optimization measures,such as the provision of shelters,were evaluated.Results Under conditions where the nutritional needs of pregnant mice were adequately met by supplementation with egg yolk and sunflower seeds,mating of female and male IFN-γ-/-mice result ed in a litter size of five to eight IFN-γ-/-mice,demonstrating higher efficiency compared to other breeding strategies.In addition,diet type and mating age significantly influenced female reproductive performance.When 7～9 weeks old female IFN-γ-/-mice were mated to male IFN-γ-/-mice and fed a high-protein breeding diet,litter size(6.9±1.7),weaning survival number(6.5％±2.0％)and weaning survival rate(93.2％±17.8％)were higher than those under other conditions.In addition,providing shelters to prevent fighting between breeding pairs further improved reproductive outcomes.Conclusions By adopting an optimized breeding strategy,combined with a high-protein diet,nutritional supplementation,and standardized mating age management,the breeding efficiency and stability of IFN-γ-/-mice can be significantly improved.This provides a reliable animal model for related research.

Objective The purpose of the study is to breed homozygous interferon-γ knockout(IFN-γ-/-)mice and optimize the breeding strategies to achieve continuous and stable reproduction of IFN-γ-/-mice,which could be used as an ideal animal model for fundamental research.Methods Initially,heterozygous IFN-γ knockout(IFN-γ+/-)C57BL/6J mice were used as the parental generation for breeding.Subsequently,3 breeding strategies were employed using the offspring:(1)female heterozygotes mated with male heterozygotes;(2)male homozygotes mated with female heterozygotes;(3)female homozygotes mated with male homozygotes.The number and survival rate of IFN-γ-/-mice were compared across the three breeding strategies to determine the optimal breeding strategy.Under the optimal strategy,the effects of female mating age and diet type on the reproductive performance of IFN-γ-/-mice were further evaluated.Data from the first three litters of 60 IFN-γ-/-female mice,including litter size,number of weaning survivors,and weaning survival rate,were recorded and analyzed.In addition,the effects of dietary supplementation of pregnant mice and environmental optimization measures,such as the provision of shelters,were evaluated.Results Under conditions where the nutritional needs of pregnant mice were adequately met by supplementation with egg yolk and sunflower seeds,mating of female and male IFN-γ-/-mice result ed in a litter size of five to eight IFN-γ-/-mice,demonstrating higher efficiency compared to other breeding strategies.In addition,diet type and mating age significantly influenced female reproductive performance.When 7～9 weeks old female IFN-γ-/-mice were mated to male IFN-γ-/-mice and fed a high-protein breeding diet,litter size(6.9±1.7),weaning survival number(6.5％±2.0％)and weaning survival rate(93.2％±17.8％)were higher than those under other conditions.In addition,providing shelters to prevent fighting between breeding pairs further improved reproductive outcomes.Conclusions By adopting an optimized breeding strategy,combined with a high-protein diet,nutritional supplementation,and standardized mating age management,the breeding efficiency and stability of IFN-γ-/-mice can be significantly improved.This provides a reliable animal model for related research.

Objective:To construct a combined radiomics model based on preoperative enhanced computed tomography (CT) examination for predicting microvascular invasion (MVI) in hepatocellular carcinoma (HCC), and provide biological explanations for the radiomics model.Methods:The retrospective cohort study was conducted. The messenger RNA (mRNA) of 424 HCC patients, the clinicopathological data of 39 HCC patients entered into the Cancer Genome Atlas database from its establishment until January 2023, and the clinicopathological data of 53 HCC patients who were admitted to the Gansu Provincial People′s Hospital from January 2020 to January 2023 were collected. The 92 HCC patients were randomly divided into a training dataset of 64 cases and a test dataset of 28 cases with a ratio of 7∶3 based on a random number table method. The CT images of patients in the arterial phase and portal venous phase as well as the corresponding clinical data were analyzed. The 3Dslicer software (version 5.0.3) was used to register the CT images in the arterial phase and portal venous phase and delineate the three-dimensional regions of interest. The original images were preprocessed and the corresponding features were extracted by the open-source software FAE (version 0.5.5). After selecting features using the Least Absolute Shrinkage and Selection Operator, the radiomics model was constructed and the radiomics score (R-score) was calculated. The nomogram was constructed by integrating clinical parameters, imaging features and R-score based on Logistic regression. The gene modules related to radiomics model were obtained and subjected to enrichment analysis by conducting weighted gene co-expression network analysis and correlation analysis. Observation indicators: (1) comparison of clinical characteristics of patients with different MVI properties; (2) establishment of MVI risk model; (3) evaluation of MVI risk model; (4) clustering of gene modules; (5) functional enrichment of feature-correlated gene modules. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Comparison of count data was conducted using the chi-square test. The intra-/inter-class correlation coefficient (ICC) was used to assess the inter-observer consistency of radiomics feature extracted by different observers. ICC >0.75 indicated a good consistency in feature extraction. The Logistic regression model was used for univariate and multivariate analyses. The receiver operating characteristic curve was drawn, and the area under curve (AUC), the decision curve and the calibration curve were used to evaluate the diagnostic efficacy and clinical practicality of the model. Results:(1) Comparison of clinical characteristics of patients with different MVI properties. Of 92 HCC patients, there were 47 cases with MVI-positive and 45 cases with MVI-negative, and there were significant differences in hepatitis, tumor diameter, peritumoral enhancement, intratumoral arteries, pseudocapsule and smoothness of tumor margin between them ( χ2=5.308, 9.977, 47.370, 32.368, 21.105, 31.711, P<0.05). (2) Establishment of MVI risk model. A total of 1 781 features were extrac-ted from arterial and portal venous phases of the intratumoral and peritumoral regions. After feature dimension reduction, 8 radiomics features were selected from arterial and portal venous phases to construct the combined model. Results of multivariate analysis showed that peritumoral enhancement, intratumoral arteries, pseudocapsule, smoothness of tumor margins, and R-score were independent risk factors for MVI in patients with HCC [ hazard ratio=0.049, 0.017, 0.017, 0.021, 2.539, 95% confidence interval ( CI) as 0.005-0.446, 0.001-0.435, 0.001-0.518, 0.001-0.473, 1.220-5.283, P<0.05]. A nomogram model was constructed incorporating peritumoral enhancement, intratumoral arteries, pseudocapsule, smoothness of tumor margins, and R-score. (3) Evaluation of the MVI risk model. The AUC of radiomics model was 0.923 (95% CI as 0.887-0.944) and 0.918 (95% CI as 0.894-0.945) in the training dataset and test dataset, respectively. The AUC of nomogram model, incorpora-ting both the R-score and radiomics features, was 0.973 (95% CI as 0.954-0.988) and 0.962 (95% CI as 0.942-0.987) in the training dataset and test dataset, respectively. Results of decision curve showed that the nomogram had better clinical utility compared to the R-score. Results of calibration curve showed good consistency between the actual observed outcomes and the nomogram or the R-score. (4) Clustering of gene module. Results of weighted gene co-expression network analysis showed that 8 gene modules were obtained. (5) Functional enrichment of feature-related gene modules. Results of correlation analysis showed 4 gene modules were significantly associated with radiomics features. The radiomics features predicting of MVI may be related to pathways such as the cell cycle, neutrophil extracellular trap formation, and PPAR signaling pathway. Conclusions:The combined radiomics model based on preoperative enhanced CT imaging can predict the MVI status of HCC. By obtaining mRNA gene expression profiles associated with radiomics features, a biological interpretation of the radiomics model is provided.

Objective:To investigate the expression of long non-coding RNA (lncRNA) MTATP6P1 in melanoma and its effect on the proliferation, migration and invasion of melanoma cells by targeting miRNA-411-5p (miR-411-5p).Methods:A total of 461 samples of melanoma tissues and paracancerous tissues (>2 cm from the tumor margin) were collected from the tumor-associated lncRNA database (TANRIC database updated in July 2021), and the expression of MTATP6P1 was compared between the two groups. The bioinformatics software lncRNA Disease v2.0 was used to predict the possible binding site microRNA (miRNA) of MTATP6P1. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of MTATP6P1 in melanoma cells A-375, WM266-4, VMM5A, A2058 and normal human epidermal melanocytes PIG1; and the lowest relative expression level of cells in MTATP6P1 were divided into MTATP6P1 group (transfected with MTATP6P1 overexpression plasmid) and NC group (transfected with blank plasmid). The proliferation ability of A-375 cells was detected by using CCK-8 method; the migration ability of A-375 cells was detected by using scratch test; the invasion ability of A-375 cells was detected by using Transwell assay; the targeting relationship between MTATP6P1 and miR-411-5p was detected by using dual luciferase reporter gene assay; Western blot was used to detect the expression of ERK signaling pathway related proteins in cells.Results:The relative expression levels of MTATP6P1 in melanoma tissues and adjacent tissues were 9.82±0.58 and 11.56±0.16, respectively. The expression level of MTATP6P1 in melanoma tissues was lower than that in paracancerous tissues ( t = 9.56, P = 0.009). The relative expression levels of MTATP6P1 in normal human epidermal melanocyte PIG1 and melanoma cells A-375, WM266-4, VMM5A, and A2058 were 1.01±0.13, 0.12±0.02, 0.66±0.04, 0.39±0.07, 0.49±0.05; the relative expression level of MTATP6P1 in melanoma cells was lower than that in PIG1 cells (all P < 0.05), and then A-375 cells with the lowest relative expression level were taken to perform the subsequent experiments. The relative expression levels of MTATP6P1 in A-375 cells of MTATP6P1 group and NC group were 14.83±1.67 and 1.02±0.30, respectively ( t = 8.13, P < 0.001). After 16, 24, 32, and 40 h of culture, the proliferation ability of the cells in the MTATP6P1 group was lower than that in NC group (all P < 0.05). The scratch healing rates of A-375 cells in MTATP6P1 group and NC group were (26±7)% and (55±4)%, respectively; the scratch healing rate in MTATP6P1 group was lower than that in NC group ( t = 3.48, P = 0.009). The invasive number of A-375 cells in MTATP6P1 group and NC group were (32±12) and (116±17), respectively; the number of invasive cells in MTATP6P1 group was lower than that in NC group ( t = 4.11, P = 0.006). The results of dual luciferase reporter gene assay showed that there was a targeting relationship between MTATP6P1 and miR-411-5p. The relative expression level of miR-411-5p in A-375 cells of MTATP6P1 group and NC group was 1.04±0.16 and 5.37±0.68, respectively; the expression level of miR-411-5p in MTATP6P1 group was lower than that in NC group ( t = 6.20, P < 0.001). The expressions of ERK signaling pathway proteins p-Ras, p-Raf, p-MEK1, p-RSK, and AP-1 in A-375 cells of MTATP6P1 group were lower than those in NC group. Conclusions:MTATP6P1 inhibits the proliferation, migration and invasion of melanoma A-375 cells through targeting miR-411-5p.

This article has systematically sorted out and verified the name， origin， producing area， quality evaluation， harvesting and processing of Pruni Semen by consulting ancient materia medica， medical books， prescription books and modern literature， in order to provide a basis for the development of famous classical formulas containing Pruni Semen. The results showed that Pruni Semen， as a medicinal material， has been widely used in medical literature of past dynasties since it was collected in Shennong Bencaojing， and also included under the names such as Yuhe， Yuzi and Yuli， and aliases such as Jueli， Queli and Chexiali. The primordial plants mentioned in the past dynasties involve about 12 species of Rosaceae， but with Prunus humilis， P. japonica and P. glandulosa as mainstream varieties used in the past dynasties， while the 2020 edition of Chinese Pharmacopoeia stipulates that the basal plants are P. humilis， P. japonica and P. pedunculata. Most of the ancient records for the origin of Pruni Semen are found everywhere in high mountains， valleys and hills， modern literature records that its origin varies according to its base， for example， P. humilis and P. japonica are mainly produced in Hebei， eastern Inner Mongolia， Liaoning， Shandong and other regions of China， and P. pedunculata is mainly produced in Inner Mongolia. Modern literature summarizes its quality as faint yellow， full and fulfilling， neat and not broken， and non-oiling， and the small Pruni Semen is better than the big Pruni Semen. The ancient processing methods of Pruni Semen mainly include blanching and peeling， blanching and peeling followed by frying， and blanching and peeling followed by pounding， with the common feature of blanching and peeling. The successive editions of Chinese Pharmacopoeia stipulate that it should be pounded when used. Based on the results of the herbal textual research and the writing time of Bianzhenglu， and combined with the market survey of Pruni Semen， it is suggested that P. humilis or P. japonica should be used as the origin of Pruni Semen in Sanpiantang， and it is harvested when the fruits are ripe， the kernels are collected by removing the stones， and processed by blanching， peeling and pounding consulting the decoction method in the current edition of Chinese Pharmacopoeia.

Objective:To explore the influence of obstructive sleep apnea hypopnea syndrome (OSAHS) on nocturnal blood pressure and angina pectoris in patients undergoing cardiac artery bypass graft (CABG) .Methods:From January to December 2018, we selected 76 patients with simple CABG at Beijing Anzhen Hospital affiliated to Capital Medical University by purposive sampling. Before surgery, patients were treated with the portable sleep respiratory monitoring and bedside blood pressure monitoring. Patients were divided into group of mild or no OSAHS ( n=35) and group of moderate or severe OSAHS ( n=41) based on apnea hypopnea index (AHI) . We compared the differences in the baseline data, sleep respiratory examination indexes, nocturnal angina pectoris, blood pressure at 21∶00, 3∶00 and 6∶00 among patients between two groups. Results:Among CABG patients with moderate or severe OSAHS, the systolic pressure at 3∶00 and 6∶00 was (115.8±12.5) mmHg and (156.8±17.5) mmHg respectively all higher than those among CABG patients with mild or no OSAHS with statistical differences ( P<0.05) ; the incidence of nocturnal angina pectoris was 29.27% (12/41) higher than that [2.86% (1/35) ]among CABG patients with mild or no OSAHS with a statistical difference ( P<0.05) . Logistic regression analysis showed that the systolic pressure at 3∶00 could affect the occurrence of nocturnal angina pectoris among CABG patients with OSAHS ( OR=1.050, 95% CI 1.003-1.100, P=0.039) ; the decrease of average oxygen saturation could also affect the occurrence of nocturnal angina pectoris among those patients ( OR=0.628, 95% CI 0.397-0.993, P=0.046) ; the differences were all statistical. Conclusions:Early morning systolic pressure of CABG patients with OSAHS raises, and OSAHS may increase the incidence of nocturnal angina pectoris among CABG patients.

Objective To investigate the relationship between DNA topoisomerase Ⅱ α (TOP2A) gene expression and clinicopathological characteristics and its significance of prognostic evaluation for patients with bladder cancer.Methods Bladder cancer gene expression profile GSE13507 (n =165) and GSE31189 (n =52) were obtained.The expression profile and clinical information of patients with bladder cancer were retrospectively analyzed,and the survival analysis was made.Gene set enrichment analysis (GSEA) was conducted to explore the related pathways which were regulated by TOP2A.Results Compared with normal bladder tissues,TOP2A was upregnlated in bladder cancer tissues (5.823 ± 1.079 vs.4.820 ± 1.129),with a statistically significant difference (t =4.336,P ＜ 0.001).The TOP2A gene expression in patients with bladder cancer was correlated with the age of patients (x2 =5.926,P =0.015),sex (x2 =6.046,P =0.014),T staging (x2 =19.484,P ＜ 0.001),N staging (x2 =9.178,P =0.002),M staging (x2 =21.142,P ＜ 0.001),tumor grade (x2 =47.005,P ＜ 0.001),and progression (x2 =11.735,P =0.001),but it was not correlated with recurrence (x2 =0.808,P =0.369).Survival analysis showed that the specific survival rate in the 100 months of TOP2A gene high expression group and low expression group had a statistically significant difference (66.59％ vs.87.95％,x2 =15.820,P ＜ 0.001).The median overall survival time of TOP2A gene high expression group and low expression group were 51.77 months and 134.97 months respectively,with a statistically significant difference (x2 =11.280,P =0.008).The results of GSEA indicated that TOP2A could regulate gene sets related with several pathways like MYC-V1 signaling (P =0.035,FDR =0.132),MYC-V2 signaling (P =0.012,FDR =0.058),E2F signaling (P ＜ 0.001,FDR =0.006) and G2M checkpoint (P =0.006,FDR =0.044).Conclusion The TOP2A gene expression is closely related with clinicopathological characteristics of patients with bladder cancer.TOP2A may function as a potential marker of prognosis for patients with bladder cancer.

Objective To investigate the MSCT manifestations of hepatic fat-poor perivascular epithelioid cell tumor (PEComa). Methods CT and pathological findings of 8 patients with hepatic fat-poor PEComa confirmed by surgery were assessed retrospectively.Results 8 cases had solitary lesion,all lesions with round or round-like shape.The largest diameter ranged 20-110 mm.The fat density was not measured by CT scan.6 lesions composed of solid component,and solid part showed obviously enhancement on arterial phase.On portal venous phase and parenchymal phase,the tumors showed equal or low or slightly high density.2 lesions showed cystic necrosis,peripheral enhancement on arterial phase,1 lesion showed continuous enhancement and the enhancement degree increased,and another lesion showed reduced enhancement.All lesions had tortuous vascular in the center of lesions or at the peripheral.Immunohistochemistry examinations showed that HMB45,Melan-A and SMA were positive in all cases,CD31,CD34 and S-100 expressed positive in partial cases. Conclusion The CT findings of hepatic fat-poor PEComa are lack of specificity.When the enhancement and clearance pattern of liver mass is similar to hepatocellular carcinoma or adenoma,the clinical history does not support the diagnosis,may consider the possibility of PEComa when tortuous vascular in the center of lesions or at the peripheral on arterial phase.

Objective To explore the CT characteristics of mycobacteria intracellulare lung disease.Methods CT imaging data of 37 patients with mycobacteria intracellulare lung disease in our hospital were analyzed retrospectively.Results The main CT signs of mycobacteria intracellulare lung disease included centrilobular nodules (97.3%), bronchiectasis (73.0%) and patchy consolidation (54.1%).Fibrous lesion(43.2%), diameter≥1 cm nodules(35.1%)and thin-wall cavity(29.7%) were also common found in patients.Thick-wall cavity and pleural effusion were not common.The typical CT manifestation was bronchiectasis accompany by the centrilobular nodules occurred in the right middle lobe and (or) left lingular lobe.Conclusion CT manifestations of mycobacteria intracellulare lung disease have some characteristics, and CT examination has a certain value in the diagnosis of this disease.

Objective To explore the clinical and MSCT manifestations of nontuberculous mycobacteria (NTM) lung diseases.Methods Totally 102 patients with proved NTM lung diseases (NTM group) and 102 patients with pulmonary tuberculosis (TB group) were included in the study.MSCT image and clinical data of patients were retrospectively analyzed.The t/x2 test were used to analyze the differences of clinical and imaging findings between two groups.Results The main clinical symptoms of NTM group were cough,expectoration,hemoptysis and shortness of breath after activity,which had no significant differences between two groups (all P＞ 0.05).NTM lung diseases patients often associated with chronic lung diseases such as pulmonary tuberculosis,chronic obstructive pulmonary disease,pulmonary heart disease.The differences were significant between two groups (all P＜0.05).The main CT manifestations of NTM lung diseases included centrilobular nodules (89/102,87.25％),bronchiectasis (67/102,65.69％) and patchy consolidation (64/102,62.75％).Secondly,fiber cable disease,thin-wall cavities and pleural incrassation were common found.The detection rate of centrilobular nodules,bronchiectasis and thin-wall cavities in NTM group were significantly higher than those in TB group (x2 =3.995,22.675,12.823,respectively,all P＜0.05).Bronchiectasis were often found in the right middle lobe and/or left lingula lobe.Conclusion NTM lung diseases patients often associate with chronic lung disease.The CT manifestations of NTM lung diseases have certain characteristics.Especially when the bronchiectasis occurred in the right middle lobe and/or left lingular lobe and accompany by the centrilobular nodules,thin-wall cavity and antituberculous therapy being invalid,NTM lung diseases should be considered.