In this study, lipopolysaccharide(LPS), ovalbumin(OVA), and compound 48/80(C48/80) were administered to establish non-infectious pneumonia models under simulated clinical conditions, and the correlation between their pathological characteristics and traditional Chinese medicine(TCM) syndromes was compared, providing the basis for the selection of appropriate animal models for TCM efficacy evaluation. An acute pneumonia model was established by nasal instillation of LPS combined with intraperitoneal injection for intensive stimulation. Three doses of OVA mixed with aluminum hydroxide adjuvant were injected intraperitoneally on days one, three, and five and OVA was administered via endotracheal drip for excitation on days 14-18 to establish an OVA-induced allergic pneumonia model. A single intravenous injection of three doses of C48/80 was adopted to establish a C48/80-induced pneumonia model. By detecting the changes in peripheral blood leukocyte classification, lung tissue and plasma cytokines, immunoglobulins(Ig), histamine levels, and arachidonic acid metabolites, the multi-dimensional analysis was carried out based on pathological evaluation. The results showed that the three models could cause pulmonary edema, increased wet weight in the lung, and obvious exudative inflammation in lung tissue pathology, especially for LPS. A number of pyrogenic cytokines, inclading interleukin(IL)-6, interferon(IFN)-γ, IL-1β, and IL-4 were significantly elevated in the LPS pneumonia model. Significantly increased levels of prostacyclin analogs such as prostaglandin E2(PGE2) and PGD2, which cause increased vascular permeability, and neutrophils in peripheral blood were significantly elevated. The model could partly reflect the clinical characteristics of phlegm heat accumulating in the lung or dampness toxin obstructing the lung. The OVA model showed that the sensitization mediators IgE and leukotriene E4(LTE4) were increased, and the anti-inflammatory prostacyclin 6-keto-PGF2α was decreased. Immune cells(lymphocytes and monocytes) were decreased, and inflammatory cells(neutrophils and basophils) were increased, reflecting the characteristics of "deficiency", "phlegm", or "dampness". Lymphocytes, monocytes, and basophils were significantly increased in the C48/80 model. The phenotype of the model was that the content of histamine, a large number of prostacyclins(6-keto-PGE1, PGF2α, 15-keto-PGF2α, 6-keto-PGF1α, 13,14-D-15-keto-PGE2, PGD2, PGE2, and PGH2), LTE4, and 5-hydroxyeicosatetraenoic acid(5S-HETE) was significantly increased, and these indicators were associated with vascular expansion and increased vascular permeability. The pyrogenic inflammatory cytokines were not increased. The C48/80 model reflected the characteristics of cold and damp accumulation. In the study, three non-infectious pneumonia models were constructed. The LPS model exhibited neutrophil infiltration and elevated inflammatory factors, which was suitable for the efficacy study of TCM for clearing heat, detoxifying, removing dampness, and eliminating phlegm. The OVA model, which took allergic inflammation as an index, was suitable for the efficacy study of Yiqi Gubiao formulas. The C48/80 model exhibited increased vasoactive substances(histamine, PGs, and LTE4), which was suitable for the efficacy study and evaluation of TCM for warming the lung, dispersing cold, drying dampness, and resolving phlegm. The study provides a theoretical basis for model selection for the efficacy evaluation of TCM in the treatment of pneumonia.
Animals ; Disease Models, Animal ; Mice ; Pneumonia/genetics* ; Medicine, Chinese Traditional ; Male ; Humans ; Cytokines/immunology* ; Female ; Lipopolysaccharides/adverse effects* ; Lung/drug effects* ; Drugs, Chinese Herbal ; Ovalbumin ; Mice, Inbred BALB C

Celastrol and wilforlide A are the main active triterpenoids of the traditional Chinese medicine Lei Gong Teng, which have anti-tumour, anti-inflammatory and immunosuppressive activities, and are the material basis for the clinical efficacy of Lei Gong Teng-related Chinese medicinal preparations. By analysing the biosynthetic pathway of active ingredients, optimizing genetic elements and utilizing "cell factory" to produce triterpenoids heterologously will be an effective way to obtain from Tripterygium wilfordii in the future in a "low-cost and high-efficient" manner. CYP712Ks are the first cytochrome P450s involved in the skeleton modification of friedelane-type and oleanane-type triterpenoids in T. wilfordii, and they can catalyse the generation of polpunonic acid from friedelin and 3-epi-katonic acid from β-amyrin by carboxylation at C-29 position. In this study, four multifunctional TwCYP712K1/2/3/5 were used to clarify the catalytic function and substrate selection preference using in vivo functional characterization in Saccharomyces cerevisiae. The spatial structure of the protein-substrate binding was clarified through homology modeling and molecular docking, and then the differential amino acids in the active pocket of the protein were mutated to clarify the crucial amino acids determining the catalytic function and the selection of substrate structure. A total of 63 mutant elements were constructed, and the amino acid sites affecting the carboxylation function of TwCYP712Ks were analyzed. In particular, the key amino acids affecting the substrate selectivity of TwCYP712K2 towards oleanane-type and friedelane-type triterpenoids were revealed and the TwCYP712K2^F127I and TwCYP712K2^A227T mutants would result in a reversal of the product ratio. In conclusion, four proteins of TwCYP712Ks were semi-rationally designed by homologous protein alignment and mutual mutation to elucidate multiple amino acid sites determining the catalytic function of the proteins, and a series of activity-enhancing or altering mutants were obtained, which provide abundant catalytic elements for the biosynthesis of active triterpenoids from T. wilfordii, and the mechanism of carboxylation in the C-29 position was initially elucidated.

Objective:To investigate the value of multi-parametric analysis based on dual-layer detector spectral CT (DLCT) in predicting the initial recurrence risk for papillary thyroid carcinoma (PTC).Methods:From November 2021 to October 2022, 102 PTC patients confirmed by pathology were retrospectively collected at the First Affiliated Hospital of Nanjing Medical University in this cross-sectional study. There were 25 males and 77 females, with an age of (42±13) years old. The initial recurrence risk assessment for PTC patients was categorized into a low-risk group (75 cases) and an intermediate-high-risk group (27 cases). Clinical data, including age, gender, body mass index, history of nodular goiter, history of Hashimoto thyroiditis, and preoperative thyroid function, were collected. Tumor morphological features, including size, location, shape, aspect ratio, the degree of thyroid capsule contact, calcification, and cystic change, were evaluated. Quantitative DLCT parameters, including iodine concentration (IC), standardized iodine concentration (NIC), effective atomic number (Z eff), standardized effective atomic number (NZ eff), electronic density (ED), CT values under different energy levels (40-200 keV, 30 keV intervals) and slope of energy spectrum curve (λ HU) both in the arterial and venous phase were measured. The differences in clinical, morphological features, and spectral CT quantitative parameters between the two groups were compared using independent sample ttest, Mann-Whitney U test, or χ2 test. Multivariate logistic regression analyses were used to construct three models based on clinical and morphological features, quantitative DLCT parameters and their combination, respectively. The receiver operating characteristic curve was used to evaluate the predictive performance of these models for the initial recurrence risk of PTC patients, and the area under the curve (AUC) was compared using the DeLong test. Results:Significant differences were found in gender, lesion long diameter, lesion short diameter and calcification between the low-risk group and intermediate-high-risk groups ( P<0.05). The arterial phase IC, arterial phase Z eff, arterial phase λ HU, arterial phase CT 40 keV, venous phase NIC and venous phase NZ eff in intermediate-high-risk group were significantly lower than those in the low-risk group ( P<0.05). The logistic regression analysis revealed that the clinical model included gender ( OR=2.895, 95% CI 1.047-8.002, P=0.040) and lesion long diameter ( OR=1.142, 95% CI 1.042-1.251, P=0.004), with an AUC of 0.720, sensitivity of 63.0%, and specificity of 78.7% in predicting the initial recurrence risk of PTC patients. The DLCT quantitative parameter model included arterial phase IC ( OR=0.580, 95% CI 0.370-0.908, P=0.017), venous phase NIC ( OR=0.077, 95% CI 0.011-0.536, P=0.010), and venous phase NZ eff ( OR=0.002, 95% CI 0.001-0.103, P=0.009), with an AUC of 0.774, sensitivity of 71.9%, and specificity of 70.0%. The AUC of the combined model was 0.857, with a sensitivity of 74.1%, and specificity of 88.0%, outperforming the clinical model ( Z=2.92, P=0.004) and the DLCT quantitative parameter model ( Z=2.07, P=0.046). Conclusion:Multi-parametric analysis based on DLCT can help predict the initial recurrence risk for PTC, and combining it with clinical and morphological features, the predictive accuracy can be improved.

Objective To analyze the demographic data,CT characteristics,treatment methods and follow-up results of patients with spontaneous isolated dissection of celiac artery(SIDCA)and to explore the selection of treatment strategy for SIDCA.Methods Medical records of 31 patients diagnosed as SIDCA were selected.The patients were divided into 2 groups,symptomatic group and asymptotic group.Demographic data,morbidity season,CT characteristics(type,distance of the entry site from the origin of the artery,dissection length,compression rate of the true lumen,branches involvement,organ ischemia,coexisting vasculopathy),treatment methods,and follow-up results were analyzed.Results Thirty-one patients were identified by CT contrast scan,22 were symptomatic and 9 were asymptomatic.22 patients were first diagnosed in cooler season,compared with 9 patients in warmer season(22 vs 9,Fisher's exact test,P=0.029).The difference between symptomatic and asymptomatic groups about branches involvement in patients was significant(8/14 vs 0/9,Fisher's exact test,P=0.007).Treatment included observation in 24,endovascular intervention in 6 patients and surgical repair in 1 patient.No patient required bowel resection.The mean follow-up period was 13.75 months.Except for 2 patients,the condition of the remaining patients improved or stabilized during follow-up.Conclusion The difference between symptomatic and asymptomatic groups about branches involvement in patients is significant.Initial conservative treatment may be adequate for patients without end organ malperfusion or aneurysm formation or aneurysm rupture in SIDCA.Additionally,during the cooler season,visceral artery should be observed carefully on abdominal contrast CT,especially in the patient with abdominal pain,to avoid misdiagnosis.

Background Phenolic compounds may adversely affect human health, but the current relevant studies are mostly limited to the impact of single phenolic compound exposure on human health, and there is still a lack of studies on the population-based association between combined exposure to multiple common phenolic compounds and dyslipidemia. Objective To explore the association of phenolic compound combined exposure and dyslipidemia based on principal component analysis-random forest (PCA-RF) strategy. Methods The data were from the National Health and Nutrition Examination Survey (2013–2016). A total of 1301 adult residents aged ≥ 20 years with complete information on demographics and lifestyle, urine phenol concentrations (bisphenol A, bisphenol F, bisphenol S, triclocarban, benzophenone, and triclosan), and serum concentrations of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were included in this study. The concentrations of six urinary phenolic compounds were determined by solid phase extraction coupled with high performance liquid chromatography and tandem mass spectrometry, and the lipid indicators were determined by enzymatic methods. Principal component analysis combined with random forest model was used for model construction. First, principal component analysis was performed on 18 original variables including 6 phenolic compounds and 12 basic characteristic indicators, and then random forest model was established with dyslipidemia and its four evaluation indicators as dependent variables and the extracted principal components as independent variables, respectively. Results The PCA-RF analysis showed that bisphenol A, bisphenol F, and benzophenone may be important factors for dyslipidemia in the study subjects; bisphenol A, bisphenol F, and triclosan may be important factors for TC level in the study subjects; bisphenol A, bisphenol F, triclocarban, and benzophenone may be important factors for TG level in the study subjects; bisphenol A may be an important factor for LDL-C level in the study subjects; bisphenol F and benzophenone may be important factors for HDL-C level in the study subjects. Conclusion Phenolic compound exposure may be an important risk factor for the development of dyslipidemia. PCA-RF strategy can be effectively used to explore the association between phenolic compound exposure and dyslipidemia in the population.

Objective:To investigate the value of radiomics based on three-dimensional high resolution MR vessel wall imaging (3D HRMR-VWI) for identifying culprit plaques in symptomatic patients with middle cerebral atherosclerosis.Methods:The clinical and imaging features of 117 patients (139 middle cerebral artery plaques) with cerebrovascular diseases in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from October 2018 to October 2020 were respectively reviewed. Stratified random sampling was used to divide 139 plaques into training set (97 plaques) and validation set (42 plaque) at the ratio of 7∶3. The plaques were divided into 69 culprit plaques and 70 non-culprit plaques based on plaque MR features and clinical symptoms. The clinical and imaging characteristics of culprit plaques and non-culprit plaques were compared by independent sample t-test, Mann-Whitney U test and χ 2 test, and factors with significant difference between two groups in univariate analysis were further analyzed by multivariate logistic regression to find out the independent predictors of culprit plaques. Radiomics features were extracted, screened and radiomics model was constructed using pre-and post-contrast 3D HRMR-VWI based on the training set. The combined model was constructed by combining the independent predictors and radiomics model. Receiver operating characteristic curve and area under curve (AUC) were used to evaluate the efficacy of each model, and DeLong test was used to compare the efficacy of different models. Results:Significant difference was found in intraplaque hemorrhage, lumen area of stenosis, stenosis diameter, stenosis rate, plaque burden and enhancement rate between culprit and non-culprit plaques (all P<0.05). Multivariate logistic regression analysis confirmed that only intraplaque hemorrhage was the independent predictor for culprit plaques (OR=7.045,95%CI 1.402-35.397, P=0.018). In the validation set, the AUC of the pre-contrast 3D HRMR-VWI model was lower than that of the post-contrast 3D HRMR-VWI model ( Z=-2.01, P=0.044). The AUC of pre+post-contrast 3D HRMR-VWI model was not significantly different from that of post-contrast 3D HRMR-VWI model ( Z=0.79, P=0.427). The AUC showed no significant difference between combined model and pre+post-contrast 3D HRMR-VWI model ( Z=-0.59, P>0.05). The combined model showed the best performance in predicting culprit plaques of middle cerebral artery (AUC=0.939), with the sensitivity, specificity and accuracy of 95.24%, 76.19% and 85.71%. Conclusion:Radiomics based on 3D HRMR-VWI has potential values in identifying culprit plaques in symptomatic patients with middle cerebral atherosclerosis.

新冠病毒感染疫情严重威胁着世界各国人民的生命健康。目前，对病毒感染的防治研究主要集中在抑制病毒与分子受体的结合上。AXL作为新发现的严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）受体，在协助病毒感染人体呼吸系统中发挥着重要作用，是未来临床干预的潜在靶点。本研究对已发表的单细胞测序数据进行整理和分析，发现AXL在年轻人肺细胞中的表达水平明显高于老年人。人胚肺二倍体成纤维细胞（2BS）是衰老研究的公认细胞株。本文采用2BS细胞构建复制性细胞衰老模型，发现年轻细胞中AXL的蛋白水平明显高于衰老细胞，据此推测年轻人感染的风险可能更高，需要注意防护。我们发现一种羊栖菜褐藻多糖硫酸酯组分（SFW-3）可显著下调年轻2BS细胞中AXL的表达水平，表明SFW-3具有一定的抗SARS-CoV-2感染的研究价值，同时表明2BS细胞株也可作为潜在的SARS-CoV-2体外感染模型。
Humans ; SARS-CoV-2 ; Sargassum/metabolism* ; Diploidy ; Sulfates/metabolism* ; COVID-19 ; Polysaccharides/pharmacology* ; Lung

Objective: To examine the clinical efficacy of 3 anti-reflux methods of digestive tract reconstruction after proximal gastrectomy for gastric cancer. Methods: The clinical data and follow-up data of gastric cancer patients who underwent anti-reflux reconstruction after proximal gastrectomy in 11 medical centers of China from September 2016 to August 2021 were retrospectively collected, including 273 males and 65 females, aging of (63±10) years (range: 28 to 91 years). Among them, 159 cases were performed with gastric tube anastomosis (GTA), 107 cases with double tract reconstruction (DTR), and 72 cases with double-flap technique (DFT), respectively. The duration of operation, length of postoperative hospital stay and early postoperative complications (referring to Clavien-Dindo classification) of different anti-reflux reconstruction methods were assessed. Body mass index, hemoglobin and albumin were used to reflect postoperative nutritional status. Reflux esophagitis was graded according to Los Angeles criteria based on the routinely gastroscopy within 12 months after surgery. The postoperative quality of life (QoL) was evaluated by Visick score system. The ANOVA analysis, Kruskal-Wallis rank sum test, χ² test and Fisher's exact test were used for comparison between multiple groups, and further comparison among groups were performed with LSD, Tamhane's test or Bonferroni corrected χ² test. The mixed effect model was used to compare the trends of Body mass index, hemoglobin and albumin over time among different groups. Results: The operation time of DFT was significantly longer than that of GTA and DTR ((352±63) minutes vs. (221±66) minutes, (352±63) minutes vs. (234±61) minutes, both P<0.01). The incidence of early complications with Clavien-Dindo grade Ⅱ to Ⅴ in GTA, DFT and DTR groups was 17.0% (27/159), 9.7% (7/72) and 10.3% (11/107), respectively, without significant difference among these three groups (χ²=3.51, P=0.173). Body mass index decreased more significantly in GTA than DFT group at 6 and 12 months after surgery (mean difference=1.721 kg/m², P<0.01; mean difference=2.429 kg/m², P<0.01). body mass index decreased significantly in DTR compared with DFT at 12 months after surgery (mean difference=1.319 kg/m², P=0.027). There was no significant difference in hemoglobin or albumin fluctuation between different reconstruction methods perioperative. The incidence of reflux esophagitis one year after surgery in DTR group was 12.9% (4/31), which was lower than that in DFT (45.9% (17/37), χ²=8.63, P=0.003). Follow-up of postoperative quality of life showed the incidence of Visick grade 2 to 4 in DFT group was lower than that in GTA group (10.4% (7/67) vs. 34.6% (27/78), χ²=11.70, P=0.018), while there was no significant difference between DFT and DTR group (10.4% (7/67) vs. 22.2% (8/36, P>0.05). Conclusions: Compared with GTA and DTR, DFT is more time-consuming, but there is no significant difference in early complications among three methods. DFT reconstruction is more conducive to maintain postoperative nutritional status and improve QoL, especially compared with GTA. The risk of reflux esophagitis after DTR reconstruction is lower than that of DFT.
Aged ; Albumins ; Esophagitis, Peptic/surgery* ; Female ; Gastrectomy/methods* ; Hemoglobins ; Humans ; Male ; Middle Aged ; Quality of Life ; Retrospective Studies ; Stomach Neoplasms/surgery*

Objective: To investigate the relationship between body mass index (BMI) and sexual development in Chinese children. Methods: A nationwide multicenter and population-based large cross-sectional study was conducted in 13 provinces, autonomous regions and municipalities of China from January 2017 to December 2018. Data on sex, age, height, weight were collected, BMI was calculated and sexual characteristics were analyzed. The subjects were divided into four groups based on age, including ages 3-<6 years, 6-<10 years, 10-<15 years and 15-<18 years. Multiple Logistic regression models were used for evaluating the associations of BMI with sexual development in children. Dichotomous Logistic regression was used to compare the differences in the distribution of early and non-early puberty among normal weight, overweight and obese groups. Curves were drawn to analyze the relationship between the percentage of early puberty and BMI distribution in girls and boys at different Tanner stages. Results: A total of 208 179 healthy children (96 471 girls and 111 708 boys) were enrolled in this study. The OR values of B2, B3 and B4+ in overweight girls were 1.72 (95%CI: 1.56-1.89), 3.19 (95%CI: 2.86-3.57), 7.14 (95%CI: 6.33-8.05) and in obese girls were 2.05 (95%CI: 1.88-2.24), 4.98 (95%CI: 4.49-5.53), 11.21 (95%CI: 9.98-12.59), respectively; while the OR values of G2, G3, G4+ in overweight boys were 1.27 (95%CI: 1.17-1.38), 1.52 (95%CI: 1.36-1.70), 1.88 (95%CI: 1.66-2.14) and in obese boys were 1.27 (95%CI: 1.17-1.37), 1.59 (95%CI: 1.43-1.78), and 1.93 (95%CI: 1.70-2.18) (compared with normal weight Tanner 1 group,all P<0.01). Analysis in different age groups found that OR values of obese girls at B2 stage and boys at G2 stage were 2.02 (95%CI: 1.06-3.86) and 2.32 (95%CI:1.05-5.12) in preschool children aged 3-<6 years, respectively (both P<0.05). And in the age group of 6-10 years, overweight girls had a 5.45-fold risk and obese girls had a 12.54-fold risk of B3 stage compared to girls with normal BMI. Compared with normal weight children, the risk of early puberty was 2.67 times higher in overweight girls, 3.63 times higher in obese girls, and 1.22 times higher in overweight boys, 1.35 times higher in obese boys (all P<0.01). Among the children at each Tanner stages, the percentage of early puberty increased with the increase of BMI, from 5.7% (80/1 397), 16.1% (48/299), 13.8% (27/195) to 25.7% (198/769), 65.1% (209/321), 65.4% (157/240) in girls aged 8-<9, 10-<11 and 11-<12 years, and 6.6% (34/513), 18.7% (51/273), 21.6% (57/264) to 13.3% (96/722), 46.4% (140/302), 47.5% (105/221) in boys aged 9-<10, 12-<13 and 13-<14 years, respectively. Conclusions: BMI is positively correlated with sexual development in both Chinese boys and girls, and the correlation is stronger in girls. Obesity is a risk factor for precocious puberty in preschool children aged 3-<6 years, and 6-<10 years of age is a high risk period for early development in obese girls.
Adolescent ; Body Mass Index ; Child ; Child, Preschool ; China/epidemiology* ; Cross-Sectional Studies ; Female ; Humans ; Male ; Obesity/epidemiology* ; Overweight/epidemiology* ; Puberty ; Puberty, Precocious ; Sexual Development

Objective: To explore the molecular mechanism of cleft palate in mice induced by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD). Methods: The pregnant mice were randomly divided into TCDD-treated group (n=42) and control group (n=42). TCDD-treated group was given by gavage a single dose of TCDD (64 μg/kg) at 8: 00 AM on gestation day 10 (GD10) and the control group was given by gavage the isopyknic corn oil. At GD13-GD15, the fetal mice palate development was observed by HE staining. The mouse embryonic palatal mesenchymal cell proliferation was detected by 5-bromo-2-deoxyuridine (BrdU) immunofluorescence. The localization and expression of maternally expressed gene3 (MEG3) in mouse embryonic palatal mesenchymal cells was detected by situ hybridization and real-time PCR (RT-PCR). The key protein expressions of transforming growth factor-β (TGF-β)/Smad signaling pathway in mouse embryonic palatal mesenchyme were analyzed by Western blotting. The interaction of MEG3 and TGF-β receptor Ⅰ (TGF-βRⅠ) was examined by RNA binding protein immunoprecipitation (RIP). Results: At GD13 and GD14, compared with the control group, the ratio of BrdU-positive cells in the palatal mesenchyme of TCDD-treated fetuses decreased significantly (GD13, t=6.66, P=0.003; GD14, t=6.56, P=0.003). However, at GD15, the ratio of BrdU-positive cells was significantly increased (t=-5.98, P=0.004). MEG3 was mainly expressed in the nuclei of fetal mouse palatal mesenchymal cells, and the expression of MEG3 in TCDD group was significantly increased at GD13, GD14 and GD15(GD13, t=39.28, P=0.012; GD14, t=18.75, P=0.042; GD15, t=28.36, P=0.045). At GD14, TCDD decreased the levels of p-Smad2 and Smad4 in embryonic palate mesenchymal cells (p-Smad2, t=9.48, P=0.001;Smad4, t=63.10, P=0.001), whereas the expression of Smad7 was significantly increased at GD14 (t=30.77, P<0.001). The results of the RIP experiment showed that the amount of TGF-βRⅠ-bound MEG3 in mouse embryonic palatal mesenchymal cells in the TCDD group (23.940±1.301) was higher than that in the control group (8.537±1.523)(t=24.55, P<0.001). Conclusions: MEG3 is involved in the suppression of mouse embryonic palatal mesenchymal cell proliferation, functioning at least in part via interacting with the TGF-βRⅠ protein and thereby suppressing Smad signaling in the context of TCDD induced cleft palate.
Animals ; Bromodeoxyuridine ; Cleft Palate/genetics* ; Female ; Mice ; Mice, Inbred C57BL ; Palate/metabolism* ; Polychlorinated Dibenzodioxins/toxicity* ; Pregnancy