Objective:To establish an immortalized human keloid fibroblasts(KFbs) cell line and identify its characteristics and functions.Methods:The specimen was obtained from a 32-year-old female patient who underwent surgical resection of an earlobe keloid at Peking University Third Hospital in November 2019. The keloid tissue obtained was removed from the subcutaneous fat and epidermis. It was then separated and cultured using the tissue sticking method to obtain primary KFbs, which were passaged using the trypsin digestion method. After the primary KFbs were infected with an SV40 lentivirus, purified by puromycin, and passaged, a human KFbs cell line was established. Chromosomal karyotype analysis, short tandem repeat (STR) profiling, and gender gene detection were conducted to identify the primary KFbs and the cell line. The CCK-8 method was used to assess the proliferation ability of the cells. Quantitative real-time PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression levels of specific genes (PGK1, ENO1, LDHA, GLUT1, TGF-β1, COL1, COL3, FN). The comparative analysis of relevant data between primary KFbs and the cell line was conducted using t-test, and P<0.05 indicated statistical significance. Results:The morphology of both the primary KFbs and the cell line was typically spindle-shaped. The cell line morphology was basically similar to that of the primary KFbs, which were continuously cultured and passaged for 20 generations. The gender gene(Amelogenin) detection showed both were females. The chromosome karyotyping of the primary KFbs and cell line was satisfactory, maintaining the fundamental characteristics of normal cells without undergoing malignant transformation. The STR identification results showed that no multiple alleles were found in the cell line, indicating a normal cell genotype. Furthermore, the cell line did not match any entries in known cell databases. After 24, 48, and 72 hours of culture, the proliferation ability of the cell line increased by 76.1%, 125.8%, and 60.3% compared to primary KFbs. The proliferation rates of the cell line were significantly faster than those of primary KFbs ( P<0.05). The mRNA and protein expression levels of the aforementioned genes in the cell line showed no significant changes compared to the primary KFbs ( P>0.05). Conclusion:An immortalized human KFbs cell line was successfully established, showing no significant changes in morphology, characterization, and function, while exhibiting a faster proliferation rate compared to that of primary KFbs.

Objective:To establish an immortalized human keloid fibroblasts(KFbs) cell line and identify its characteristics and functions.Methods:The specimen was obtained from a 32-year-old female patient who underwent surgical resection of an earlobe keloid at Peking University Third Hospital in November 2019. The keloid tissue obtained was removed from the subcutaneous fat and epidermis. It was then separated and cultured using the tissue sticking method to obtain primary KFbs, which were passaged using the trypsin digestion method. After the primary KFbs were infected with an SV40 lentivirus, purified by puromycin, and passaged, a human KFbs cell line was established. Chromosomal karyotype analysis, short tandem repeat (STR) profiling, and gender gene detection were conducted to identify the primary KFbs and the cell line. The CCK-8 method was used to assess the proliferation ability of the cells. Quantitative real-time PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression levels of specific genes (PGK1, ENO1, LDHA, GLUT1, TGF-β1, COL1, COL3, FN). The comparative analysis of relevant data between primary KFbs and the cell line was conducted using t-test, and P<0.05 indicated statistical significance. Results:The morphology of both the primary KFbs and the cell line was typically spindle-shaped. The cell line morphology was basically similar to that of the primary KFbs, which were continuously cultured and passaged for 20 generations. The gender gene(Amelogenin) detection showed both were females. The chromosome karyotyping of the primary KFbs and cell line was satisfactory, maintaining the fundamental characteristics of normal cells without undergoing malignant transformation. The STR identification results showed that no multiple alleles were found in the cell line, indicating a normal cell genotype. Furthermore, the cell line did not match any entries in known cell databases. After 24, 48, and 72 hours of culture, the proliferation ability of the cell line increased by 76.1%, 125.8%, and 60.3% compared to primary KFbs. The proliferation rates of the cell line were significantly faster than those of primary KFbs ( P<0.05). The mRNA and protein expression levels of the aforementioned genes in the cell line showed no significant changes compared to the primary KFbs ( P>0.05). Conclusion:An immortalized human KFbs cell line was successfully established, showing no significant changes in morphology, characterization, and function, while exhibiting a faster proliferation rate compared to that of primary KFbs.

Objective:To assess the predictive value of dosiomics in predicting the occurrence of bone marrow suppression (BMS) in patients with pelvic tumors during radiotherapy.Methods:A retrospective analysis was conducted on the clinical data and radiotherapy planning documents of 129 patients with pelvic region tumors who underwent radiotherapy at the First Affiliated Hospital of Nanjing Medical University from January 2019 to January 2023. The region of interest (ROI) was outlined for bone marrow in the pelvic region by Accu Contour software in planning CT, and the ROI was exported together with the dose distribution file. According to a stratified randomization grouping method, the patients were divided into the training set and test set in an 8 vs. 2 ratio. The dosiomic features were extracted from the ROI, and the two independent samples t-test and the least absolute shrinkage and selection operator (LASSO) algorithm was employed to identify the best predictive characteristics. Subsequently, the dosiomic scores were calculated. Clinical predictors were identified through both univariant and multivariate logistic regression analyses. Predictive models were constructed by using clinical predictors alone and combining clinical predictors and dosiomic scores. The efficacy of predictive model was assessed by plotting the receiver operating characteristic (ROC) curve and evaluating its performance through the area under the ROC curve (AUC), the calibration curve, and decision curve analysis (DCA). Results:Fourteen dosiomic features that showed a strong correlation with the occurrence of BMS were screened and utilized to calculate the dosiomic scores. Based on both univariant and multivariate logistic regression analyses, chemotherapy, planning target volume (PTV) and V 5 Gy were identified as clinical predictors. According to the combined model, the AUC values for the training set and test set were 0.911 and 0.868, surpassing those of the clinical model (AUC=0.878 and 0.824). Furthermore, the analysis of both the calibration curve and DCA suggested that the combined model had higher calibration and net clinical benefit. Conclusion:The combined model has a high diagnostic value for predicting BMS in patients with pelvic tumors during radiotherapy.

Objective:To observe the detrimental effect of airborne black carbon suspension solution of different concentrations on the tear film function of mice.Methods:Twenty-eight SPF-grade male BALB/c mice, aged 6-8 weeks, were randomly divided into four groups, 0.5 mg/ml group, 1 mg/ml group, 5 mg/ml group, and control group, with 7 mice in each group.The right eyes of mice were dropped by 4 μl of 0.5 mg/ml, 1 mg/ml, 5 mg/ml black carbon suspension, or phosphate buffer solution, 3 times a day according to grouping.Tear volume, tear break-up time (TBUT), corneal fluorescein staining (CFS), and conjunctival congestion were assessed before treatment and 4, 7, 10, and 14 days after treatment.The use and care of experimental animals complied with the Regulations for Administration of Laboratory Animals in Peking Union Medical College Hospital (No.XHDW-2022-053).Results:At 14 days after treatment, the tear volumes of 0.5 mg/ml group, 1 mg/ml group, 5 mg/ml group and control group were (2.74±0.74), (2.73±0.76), (2.31±0.67), and (5.31±0.36)mm, respectively.TBUT of the four groups were (4.87±0.28), (4.00±0.76), (3.23±0.43), and (6.22±0.22)seconds, respectively.CFS of the four groups were 4(3, 4), 5(5, 6), 7(7, 8) and 0(0, 1) points, respectively.Conjunctival congestion grades of the four groups were 2(2, 3), 2(2, 3), 3(2, 3) and 0(0, 1), respectively.There were statistically significant differences in tear volume among the four groups at different time points ( Fgroup=83.325, P<0.001; Ftime=86.551, P<0.001; Finteraction=5.181, P<0.001). Before and at each time point after treatment, tear volumes were significantly lower in 0.5 mg/ml group, 1 mg/ml group, and 5 mg/ml group than in control group, and tear volumes in 0.5 mg/ml group, 1 mg/ml group, and 5 mg/ml group were significantly lower at 4, 7, 10, and 14 days after treatment than before treatment (all at P<0.05). There were statistically significant differences in TBUT among the four groups at different time points ( Fgroup=75.130, P<0.001; Ftime=56.265, P<0.001; Finteraction=6.103, P<0.001). Before and at each time point after treatment, TBUT was significantly shorter in 0.5 mg/ml group, 1 mg/ml group, and 5 mg/ml group than in control group, and the TBUT was significantly shorter in 0.5 mg/ml group, 1 mg/ml group, and 5 mg/ml group at 4, 7, 10, and 14 days after treatment than before treatment, shorter in 1 mg/ml group and 5 mg/ml group at 14 days after treatment than 4, 7 and 10 days after treatment (all at P<0.05). There were statistically significant differences in CFS score and conjunctival congestion grades among the four groups at different time points, but the interactions between concentration group and measurement time were not statistically significant (CFS: Hgroup=59.249, P<0.001; Htime=49.959, P<0.001; Hinteraction=15.980, P=0.192.conjunctival congestion grade: Hgroup=57.622, P<0.001; Htime=42.062, P<0.001; Hinteraction=12.565, P=0.401). Before and at each time point after treatment, the CFS scores and conjunctival congestion grades were significantly higher in 0.5 mg/ml group, 1 mg/ml group, and 5 mg/ml group than in control group, and CFS scores and conjunctival congestion grades were significantly higher in 0.5 mg/ml group, 1 mg/ml group, and 5 mg/ml group at 4, 7, 10, and 14 days after treatment than before treatment (all at P<0.05). Conclusions:The exposure of airborne black carbon on the ocular surface causes damage to tear film function and ocular surface inflammation in BALB/c mice.Within a certain concentration and time range, the tear secretion decreases, TBUT shortens, CFS and conjunctival congestion increase.

Surgical treatment of intrauterine hydrocephalus can prevent irreversible fetal brain damage through early decompression of the lateral ventricle. In 1980s, the prognosis of fetuses with hydrocephalus who received intrauterine treatment were poor due to non-specific surgical indications, lacking skilled operators, and underdeveloped imaging technology. We review the development of the surgical indications for fetal hydrocephalus in the following four stages: the introduction of surgical indications, the exclusion of extracranial malformations, the clear definition of isolated hydrocephalus, and the popularization of micro-array and gene sequencing techniques. The outcomes of fetuses with hydrocephalus who received intrauterine treatment with different selection criteria are summarized to explore the inclusion and exclusion criteria.

Objective To investigate the pathology, epidemiology of severe pneumonia in Xicheng District of Beijing. Methods From 2014 to 2020, in 3 sentinel hospitals, collected and detected the respiratory tract specimens of the severe pneumonia patients. Multiple pathogens including Stenotrophomonas maltophilia, Streptococcus pyogenes, Staphylococcus aureus, Klebsiella Pneumoniae, Haemophilus influenzae, Legionella pneumophila, Mycobacterium tuberculosis, Acinetobacter baumannii, Moraxella catarrhalis, Escherichia coli, Streptococcus pneumoniae, Pseudomonas aeruginosa, Pneumocystis,Parainfluenza virus, Bocavirus, Rhinovirus, Coronavirus, Influenza, Human metapneumovirus, Adenovirus, Respiratory syncytial virus, Enterovirus, Mycoplasma pneumoniae, Chlamydia pneumoniae were detected with PT-PCR. Analyze epidemic characteristics of the cases. Results Of the 1 247 respiratory samples cultured during the period from 2014 to 2020, 560(44.91%) are positive. The positive rates of virus(29.91%) is higher than Bacteria(20.21%). The top five pathogens were Acinetobacter baumannii(9.22%), Pseudomonas aeruginosa(8.26%), Stenotrophomonas maltophilia(7.78%), Klebsiella Pneumoniae（6.74%） and Parainfluenza virus（6.58%）. Conclusion There was a variety of pathogen in the severe pneumonia patients. Acinetobacter baumannii, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Klebsiella Pneumoniae and Parainfluenza virus were the main pathogens of respiratory infections in Xicheng district of Beijing. It is necessary to strengthen the surveillance of the disease.

Objective:To evaluate the clinical value of temporal pole and external capsule white matter hyperintensities (WMHs) on the diagnosie of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) by meta-analysis.Methods:PubMed, Cochrane, Embase, VIP database, China Biomedical Literature database, CNKI, Wanfang Data Service Platform were retrieved. The relevant literature of temporal pole and external capsule WMHs for the diagnosis of CADASIL was collected. The retrieval time limit was from the establishment of the databases to April 1, 2020. Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) was used to evaluate the quality of literature. Stata 15.1 software was used for statistical analysis. The fitted Summary Receiver Operating Characteristic (SROC) curve and combined diagnostic effect size were used to evaluate the diagnostic value of temporal pole and external capsule WMHs for CADASIL.Results:A total of 9 articles involving 10 studies were enrolled, including 880 patients. The combined sensitivities of temporal pole and external capsule WMHs for CADASIL were 0.67 (95% confidence interval [ CI] 0.54-0.78) and 0.84 (95% CI 0.72-0.91) respectively, the combined specificities were 0.64 (95% CI 0.47-0.78) and 0.44 (95% CI 0.36-0.53) respectively, the combined positive likelihood ratios were 1.9 (95% CI 1.4-2.6) and 1.5 (95% CI 1.2-1.8) respectively, the combined negative likelihood ratios were 0.51 (95% CI 0.42-0.63) and 0.37 (95% CI 0.20-0.69) respectively, the odds ratios of combined diagnosis were 4 (95% CI 3-5) and 4 (95% CI 2-9) respectively, and the area under the SROC curves were 0.71 (95% CI 0.66-0.74) and 0.62 (95% CI 0.58-0.66) respectively. Conclusions:The temporal pole and external capsule WMHs have limited diagnostic value for CADASIL, and other factors need to be comprehensively considered in the clinical diagnosis process.

BK polyomavirus-associated nephropathy (BKPyVAN) is a common cause of allograft failure. However, differentiation between BKPyVAN and type I T cell-mediated rejection (TCMR) is challenging when simian virus 40 (SV40) staining is negative, because of the similarities in histopathology. This study investigated whether donor-derived cell-free DNA (ddcfDNA) can be used to differentiate BKPyVAN. Target region capture sequencing was applied to detect the ddcfDNAs of 12 recipients with stable graft function, 22 with type I TCMR, 21 with proven BKPyVAN, and 5 with possible PyVAN. We found that urinary ddcfDNA levels were upregulated in recipients with graft injury, whereas plasma ddcfDNA levels were comparable for all groups. The median urinary concentrations and fractions of ddcfDNA in proven BKPyVAN recipients were significantly higher than those in type I TCMR recipients (10.4 vs. 6.1 ng/mL,

Objective:To develop an air particulate protective mask filter test device that can simulate the dynamics process of human breathing.Methods:The new device used two air pumps working alternately to simulate the dynamics process of human breathing. On March 4th to 17th, 2017, the new device and the traditional one-way airflow mask filtration test device were used to measure the internal and external particle levels of 39 masks of 13 models of 6 brands, and then the filtration efficiency of the mask was calculated and the test results were compared.Results:For the mask without breathing valve, there was no statistically significant difference between the filter efficiency test results of the new device and the traditional unidirectional airflow filter performance test device ( P>0.05) . For masks with breathing valves, the new device detected that three of them had lower filtration efficiency (99.50% vs 98.63%, P<0.01) . After sealing the mask breathing valve with glue, the filtering efficiency of the mask with a breathing valve detected by the new device significantly improved (98.63% vs 99.50%, P<0.01) . Conclusion:This new device can simulate the dynamic process of human exhalation and inhalation, and measure the filtration efficiency of the mask. For masks with breathing valves, the new device makes it easier to detect the decrease in the filtering efficiency of the mask caused by the breathing valve.

Due to the impact of COVID-19, the online problem-based learning (PBL) teaching mode was first implemented based on the curriculum integration of basic medicine and clinical medicine in our university in 2020. A total of 357 eight-year program clinical medicine students from three grades participated in the online PBL courses. The students have had related skills and knowledge of integration curriculum, and the teachers have carried out sufficient analysis of learning situation, preparation of software and hardware, as well as teaching design. Teachers and students has communicated with each other smoothly and cooperated closely through the Tencent Conference software platform, therefore this teaching mode has gained better feedback among students, with higher satisfaction of courses and significantly improved students' learning enthusiasm. The curriculum integration has laid the foundation for the case design and application of online PBL teaching, while the success of online PBL teaching has also reflected the original intention of curriculum integration reform.