ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.

ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.

Objective:To establish the reference values and growth curves of skeletal muscle mass among children in the Nanjing area.Methods:A cross-sectional study was conducted with children who underwent physical examination at the Department of Child Health Care, Children′s Hospital of Nanjing Medical University from 2020 January to 2022 September. Their height, weight, body fat mass and skeletal muscle mass were measured. Body mass index, percentage of body fat mass, percentage of skeletal muscle mass, relative skeletal muscle mass index and the ratio of skeletal muscle to body fat were calculated. The associations between skeletal muscle mass indices and physical measurements index were analyzed through the Spearman correlation test. The Mann-Kendall test was used to assess the trend for skeletal muscle mass. Generalized additive models for location, scale and shape were used to construct percentile reference values and growth curves of male and female skeletal muscle mass indices at different ages.Results:A total of 32 690 children aged 4-14 years were enrolled in this study, including 19 912 boys (60.91%). Skeletal muscle mass, percentage of skeletal muscle mass, relative skeletal muscle mass index and the ratio of skeletal muscle to body fat of boys and girls was 11.10 (8.40, 14.90) and 10.30 (7.90, 13.20) kg, 40.36% (37.01%, 43.13%) and 39.38% (36.43%, 41.88%), 6.70 (6.07, 7.52) and 6.33 (5.79, 7.00), 2.39 (1.46, 3.47) and 2.14 (1.45, 3.00) kg/m 2, respectively. Skeletal muscle mass of both boys and girls was all positively associated with weight ( r=0.97, 0.96), body mass index ( r=0.68, 0.63) and percentage of body fat mass ( r=0.40, 0.43) (all P<0.01). The reference values and growth curves showed that the percentage of skeletal muscle mass P50 ranged from 37.75%-44.61% in boys and from 36.22%-40.55% in girls. The relative skeletal muscle mass index P50 ranged from 5.80-9.68 kg/m 2 in boys and from 5.57-7.98 kg/m 2 in girls. The ratio of skeletal muscle to body fat P50 ranged from 1.86-2.67 in boys and from 1.29-2.41 in girls. There was an increasing trend with age for both boys and girls in the growth of skeletal muscle mass ( Z=4.20, 3.75, both Ptrend<0.01), and increased slightly before 9 years of age and then increased rapidly until 14 years of age in both boys and girls. Conclusions:The skeletal muscle mass indices change with age and gender during childhood. Percentile reference values for pediatric skeletal muscle mass indices can be used to evaluate the muscular growth and development in children in the Nanjing area.

Objective:To investigate the predictive value of neck circumference on cardiometabolic risk in children.Methods:This was a cross-sectional study of natural sources. As the prediction cohort, clinical data were collected from 3 443 children aged 5-14 years who underwent physical examination in the Department of Child Healthcare, Children′s Hospital of Nanjing Medical University from July 2021 to September 2022. As the validation cohort for external validation, clinical data were collected from 604 children aged 5-14 years who underwent physical examination in the Department of Child Healthcare, Children′s Hospital of Nanjing Medical University from October 2022 to March 2023. Height, weight, neck circumference, waist circumference and body composition were measured in both groups, and body mass index, neck circumference to height ratio (NHtR), waist circumference to height ratio, body fat percentage and skeletal muscle percentage were calculated. Systolic blood pressure, diastolic blood pressure, fasting blood glucose, blood lipid and uric acid and other cardiovascular and metabolic risk indicators were collected in both groups. The prediction cohort was further stratified into clustered and non-clustered groups based on the clustering of cardiometabolic risk factors (CCRF). Various variables between these 2 groups were compared using the Mann-Whitney U test. Pearson correlation and binary Logistic regression were conducted to investigate the correlations between neck circumference and cardiovascular metabolic risk factors. The accuracy of NHtR in predicting the CCRF was evaluated using the area under the curve (AUC) of receiver operating characteristic (ROC). The cutoff value was determined using the Youden index. The validation cohort was then divided into groups above and below the cutoff value, and the detection rate of CCRF between the 2 groups was compared using the χ2 test for validation .Results:In the prediction cohort of 3 443 children (2 316 boys and 1 127 girls), 1 395 (40.5%) children were overweight or obese, and 1 157 (33.6%) children had CCRF. Pearson correlation analysis revealed all significant positive correlations (all P<0.01) between neck circumference and systolic blood pressure ( r=0.47, 0.39), diastolic blood pressure ( r=0.27, 0.21), uric acid ( r=0.36, 0.30), and triglycerides ( r=0.20, 0.20) after adjusting for age in both males and females. Among both males and females, neck circumference both showed significant negative correlation (both P<0.01) with high-density lipoprotein cholesterol ( r=-0.27, -0.28), and no correlation with fasting glucose levels ( r=0.03, -0.03, both P>0.05). After adjusting for gender, age, and body fat percentage, increased body mass index, neck circumference, or waist circumference increased the risks of hypertension ( OR=1.23, 1.39, 1.07, all P<0.001), hyperuricemia ( OR=1.16, 1.23, 1.05, all P<0.001), hypertriglyceridemia ( OR=1.08, 1.16, 1.02, all P<0.01), low high-density lipoprotein cholesterol ( OR=1.10, 1.27, 1.03, all P<0.01), and the CCRF ( OR=1.51, 1.73, 1.15, all P<0.01). The areas under the ROC curves of NHtR in predicting CCRF was 0.73, with sensitivity and specificity at 0.66 and 0.71, respectively. The corresponding optimal cut-off value was 0.21. Validation with 604 children confirmed that the detection of CCRF in the NHtR≥0.21 group was 3.29 times (60.5% (112/185) vs. 18.7% (79/422), χ2=107.82, P<0.01) higher compared to the NHtR <0.21 group. Conclusions:Neck circumference is associated with cardiovascular metabolic risks such as hypertension, hyperlipidemia, hyperglycemia, and hyperuricemia in children. When the NHtR is ≥0.21, there is an increased likelihood of CCRF.

African swine fever(ASF),caused by the African swine fever virus(ASFV),is a highly contagious infectious disease of pigs.This disease has been spread rapidly in China since 2018,po-sing a huge threat to China's pig farming industry.To rapid detect the ASFV,a loop-mediated iso-thermal amplification(LAMP)combined with the disposable nucleic acid visualization test strip was established for visual detection of the nucleic acid of ASFV B646L gene.The method was easy to operate without special instruments and equipment,while it effectively avoided the disadvantage of false positives caused by aerosol contamination.The method was able to detect 1.16 copies/μL of the recombinant plasmid in 50 min at 65 ℃.In addition,the method was specific with no cross-re-action with classical swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,transmissible gastroenteritis virus.The results in this study provides a rapid,con-venient,sensitive and reliable method for early diagnosis and screening for ASFV suspected infec-tion cases.

Objective:To investigate the influence of maternal autoimmune diseases and anticoagulants, including low-molecular-weight heparin (LMWH) and aspirin, on the fetal fraction of maternal plasma cell-free DNA of non-invasive prenatal testing (NIPT).Methods:A prospective cohort study was conducted on women with singleton pregnancies receiving NIPT in the Nanjing Drum Tower Hospital from March 2021 to July 2022. NIPT was carried out using a polymerase chain reaction (PCR)-free amplification platform. In this study, four types of maternal autoimmune diseases, which were antiphospholipid syndrome, undifferentiated connective tissue disease, Sj?gren's syndrome, and systemic lupus erythematosus (SLE), and two anticoagulants, LMWH and aspirin, were studied. Univariate and multivariate linear regression models were used to analyze the factors influencing fetal fraction of maternal plasma cell-free DNA.Results:A total of 4 102 singleton pregnant women were enrolled in the prospective cohort, and 3 948 were finally included after excluding the cases with unclear dosing time of LMWH or aspirin, other autoimmune diseases, conceiving through ovulation induction alone, and having true positive or failed NIPT result. There were 96 cases with antiphospholipid syndrome, 35 with undifferentiated connective tissue disease, 34 with Sj?gren's syndrome, and 18 with SLE. A total of 108 patients only received LMWH treatment, 121 only received aspirin treatment, and 113 received both LMWH and aspirin treatment. Univariate linear regression analysis showed that maternal body mass index at blood collection ( B=－0.423), conceived by assisted reproductive technology ( B=－0.803), male fetus ( B=－0.458), undifferentiated connective tissue disease ( B=1.774), and SLE ( B=3.467) had influence on the fetal fraction (all P<0.05). Multivariate linear regression analysis showed that maternal body mass index at blood collection ( B=－0.415), conceived by assisted reproductive technology ( B=－0.585), male fetus ( B=－0.322), SLE ( B=3.347) and undifferentiated connective tissue disease ( B=1.336) were factors influencing fetal fraction (all P<0.05). Conclusions:Maternal use of LMWH or aspirin does not affect fetal fraction when performing NIPT on a PCR-free amplification platform, but undifferentiated connective tissue disease and SLE are the influencing factors. Therefore, pregnant women should be informed before the NIPT that the fetal fraction of maternal plasma cell-free DNA may be affected by maternal autoimmune diseases.

Objective To investigate the effects of different doses of T-2 toxin on the expression of cytokines cytokines and pathological changes in parental mice and their offspring. Methods One hundred female mice and 25 male mice (CD-1, SPF) were adapted for one week. After regular random mating, observation of vaginal suppository within the first 24 hours was as the 0th day of pregnancy. The pregnant rats were divided into high dose, medium dose, low dose and control groups according to body weight by a random number table(Feed: the doses of T-2 toxin were 1 200, 600, 300, and 0 μg/kg, respectively), with 16 - 18 rats in each group. The high, middle and low dose groups began to consume the poisoned feed on the 0th day of pregnancy, while the control group consumed the standard feed. After natural delivery, their offspring were continually treated the same way as their mother until the offspring reached adulthood. Serum levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), organ coefficient and pathological changes of articular cartilage were determined. Results The levels of IL-1β,IL-6 and TNF-α in the control, low, middle and high T-2 toxin groups during the pro-pregnancy of the middle-aged mice were [(219.56 ± 19.32), (136.89 ± 20.41), (210.49 ± 21.23), (207.41 ± 21.23); (192.73 ± 22.43), (136.25 ± 29.55), (187.43 ± 39.32), (232.48 ± 39.32); (1 303.02 ± 142.10), (1 072.60 ± 78.30), (1 065.03 ± 37.44), and (1 169.72 ± 104.18) ng/L], respectively. The differences between control and T-2 toxin treated groups were statistically significant (F = 17.124, 6.237, 7.670, P < 0.05). For further pairwise comparison,IL-1β and IL-6 in low dose group were significantly lower than those in control, middle and high dose groups (P < 0.05); TNF-α content in control group was significantly higher than those in low,middle and high dose groups(P<0.05).There were significant differences in the levels of IL-1β,IL-6 and TNF-α between the control group and the low,middle and high dose groups of offspring weanling mice[(142.36 ± 13.36),(113.01 ± 8.65), (102.13 ± 8.31), (123.42 ± 10.41); (109.92 ± 9.76), (100.26 ± 15.60), (85.25 ± 9.97), (100.21 ± 16.46);(1 308.45 ± 204.90), (1 248.60 ± 96.85), (1 081.09 ± 105.51), (1 204.87 ± 153.96) ng/L, F = 49.823, 10.530, 7.490, P < 0.05]. The levels of IL-1β and IL-6 in the control group were significantly higher than those in the low, middle and high dose groups(P < 0.05).The levels of TNF-α in the control group were significantly higher than those in the medium and high dose groups(P < 0.05).The levels of the three cytokines IL-1β, IL-6 and TNF-α in adult filial mice were significantly different [(69.71 ± 9.61), (61.31 ± 10.07), (63.07 ± 10.39), (58.56 ± 9.69); (172.55 ± 24.55),(146.91 ± 13.47),(151.02 ± 24.93), (157.21 ± 17.86); (1 136.87 ± 137.39), (1 002.22 ± 86.52), (987.12 ± 130.80),(1 047.21 ± 171.64)ng/L, F=4.670,5.636, 4.775, P < 0.05], the contents of the three cytokines in the poisoning groups were significantly lower than that of the control group (P < 0.05). The organ coefficients of thymus, spleen and liver in the second trimester were significantly different [(0.14 ± 0.03), (0.20 ± 0.06), (0.15 ± 0.02), (0.12 ± 0.03); (0.71 ± 0.16), (0.78 ± 0.14), (0.77 ± 0.15), (0.38 ± 0.10); (6.19 ± 0.43), (5.57 ± 0.57), (6.04 ± 0.32), (5.11 ± 0.29), F = 4.056, 11.064, 8.312, P < 0.05], and the thymus index was significantly increased in low dose group (P<0.05),spleen coefficient decreased significantly in high dose group (P < 0.05), and liver coefficients in low and high dose group were significantly decreased (P < 0.05). In the offspring, the midbrain coefficient of viscera showed significant changes [(3.45 ± 0.73), (3.11 ± 0.31), (2.98 ± 0.45), (3.04 ± 0.22), F = 7.529, P < 0.05], which was significantly decreased in the exposed rats(P<0.05).Both the mid-pregnant mice and filial mice showed varying degrees of changes in epiphyseal cartilage injury. The degree of epiphyseal cartilage injury became higher with increasing dosages of T-2 toxin in mid-pregnancy and post-weaning parental mice, and the injury was more serious in post-weaning mice. Conclusions Exposure to T-2 toxin can cause decrease of cytokines IL-1β, IL-6 and TNF-α in the blood of CD-1 pregnant and filial mice, and also cause the cartilage damage in mice, which are aggravated following increased doses of T-2 toxin and extension of exposure time.

Objective To discuss the accuracy and objectivity of student standardized patients (SSP) in objective structured clinical examination(OSCE). Methods On March 30 and 31, 2013, 168 seven-year program medical students of class 2006 and 2007 took part in OSCE. Differences in as-sessment results between SPP and those given by professional doctors at 3 SP sites(angina pectoris SSP station, acute cholecystitis SSP station, depression SSP station) were analyzed. Each site had 4 items for assessment, with a total score of 100. Scores were given in strict accordance with a set of unified scoring rules. Counting data were presented as x±s. Data were verified using t test. P<0.05 was considered statis-tically considerable. Results At angina pectoris station, respective scores of SSP and professional physi-cians were 85.2±7.1 and 85.5±6.6, P=0.688. At acute cholecystitis station, respective score of SSP and professional physicians were 89.1±5.2 and 88.2±6.2, P=0.150. At depression station, respec-tive score of SSP and professional physicians were 79.8±7.5 and 78.2±7.0, P=0.078. Conclusion There is no statistical difference between scores given by SSP and those given by the physicians in OSCE. This proves that SSP who received standardized training delivers fair and accurate results in OSCE , and therefore is recommended for future application.

Utilizing standardized patient(SP) for classroom simulation is common in current medical education. In this paper, incentive measures and combination of SP with theoretical examination, simulated people and clinical practice were proposed after in-depth analysis of advantages and disadvan-tages of using medical students as SP in terms of recruitment, training, and application. All these mea-sures were intended to promote the development of simulative medical education that in turn to cultivate students to be competent in practice.

PURPOSE: Four polymorphisms, -765G>C, -1195G>A, 8473T>C, and Val511Ala, in the cyclooxygenase-2 (COX-2) gene were identified to be associated with colorectal cancer (CRC) risk. However, the results are inconsistent. The objective of this meta-analysis was to evaluate the association between these four polymorphisms and the risk of CRC. MATERIALS AND METHODS: All eligible case-control studies published up to December 2012 on the association between the four polymorphisms of COX-2 and CRC risk were identified by searching PubMed and Web of Science. The CRC risk associated with the four polymorphisms of the COX-2 gene was estimated for each study by odds ratio (OR) together with its 95 % confidence interval (CI), respectively. RESULTS: A total of 15 case-control studies were included. Overall, no evidence has indicated that the -1195A allele, -765C allele, 8473C allele, and 511Ala allele are associated with susceptibility to CRC (-1195G>A: OR=1.11, 95 % CI: 0.82-1.51, p=0.78; -765G>C: OR=1.08, 95 % CI: 0.96-1.21, p=0.07; 8473T>C: OR=1.03, 95 % CI: 0.89-1.18, p=0.91; Val511Ala: OR=0.71, 95 % CI: 0.46-1.09, p=0.94). However, stratified analysis with ethnicity indicated that individuals with -765GC or GC/CC genotypes had an increased risk of CRC among Asian populations (GC vs. GG: OR=1.05, 95 % CI: 0.87-1.28, p=0.03; GC+CC vs. GG: OR=1.08, 95 % CI: 0.96-1.21, p=0.07). CONCLUSION: This meta-analysis indicated that -765G>C polymorphism was significantly associated with susceptibility to CRC in Asian populations.
Asian Continental Ancestry Group ; Case-Control Studies ; Colorectal Neoplasms/*genetics ; Cyclooxygenase 2/*genetics ; Genetic Predisposition to Disease/genetics ; Humans ; Polymorphism, Genetic/*genetics