Objective:To establish the fingerprint analysis method for the root of Rosa laevigata Michx from different regions by UPLC. Methods:The column was ACQUITY UPLC? Phenyl(2.1 mm × 100 mm,1.7 μm). The mobile phase consisted of methanol-water with gradient elution. The flow rate was 0. 2 ml·min-1 , the detection wavelength was 210 nm, the column temperature was 30℃, and the injection volume was 3 μl. Results:The fingerprint consisted of 15 common peaks. The range of similarity for twelve bat-ches of the root of R. laevigata Michx was 0. 489-0. 942. And the reference fingerprint of the root of R. laevigata Michx was estab-lished by UPLC. Conclusion:The fingerprint method is simple and reproducible, which can provide basis for the quality control and the medicinal resources exploration.

Objective:To create radiomics models based on abbreviated multimodal magnetic resonance imaging (MRI) for the diagnosis of breast cancer.Methods:All breast MR imaging data between Jun 2014 and Mar 2019 were retrospectively collected. Patients with pathological results of puncture or surgical resection were involved in this study. One thousand three hundred and six patients (416 benign and 890 breast cancer) were divided into training cohort ( n=702), internal validation cohort ( n=302), and external validation cohort ( n=302). All images were reduced to: the joint model group [including T2 weighted imaging (T2WI), DWI (diffusion-weighted imaging) and first contrast-enhanced sequences], non-enhanced group (T2WI and DWI) and single-phase enhanced group (first contrast-enhanced sequences). Analysis of variance (ANOVA) and least absolute shrinkage and selection operator (LASSO) were used to reduce the dimension of texture features. Three supervised machine learning algorithms (Bagging decision tree, Gaussian process, support vector machine) were used to predict benign and malignant breast lesions, and the best classifier was selected to construct breast cancer diagnosis model. Models were validated by internal and external validation cohorts. Results:The Gaussian process algorithm was chosen. The area under the curve (AUC) of the joint model and the non-enhanced model for predicting breast cancer were 0.903 and 0.893 for the training cohort, 0.893 and 0.863 for the internal validation cohort, and 0.878 and 0.864 for the external validation cohort.Conclusions:The radiomics model based on abbreviated multimodal MRI can accurately diagnose breast cancer. And the non-enhanced model can accurately diagnose breast cancer without contrast enhancement, which provides feasibility for simplifying the diagnosis process.

ObjectiveTo investigate the mechanism and effect of Qingfei Paidu decoction on transient receptor potential vanilloid-1/Transient receptor potential ankyrin1 （TRPV1/TRPA1） based on heat-sensitive channel and inflammatory response. MethodAccording to body weight， 80 8-week-old C57BL/6 mice were randomly divided into the normal group， model group， dexamethasone group （5 mg·kg^-1）， and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction （14.865， 29.73， 59.46 g·kg^-1）， with 12 mice in each group. In addition to the normal group， the other groups were administered 20 μL （1×10^-3 g·kg^-1） to each mouse by airway infusion to establish the acute lung injury （ALI） model. In the administration group， the drug was given 1 h after modeling and again after an interval of 24 h. The lung tissue was taken 36 h after modeling. Double lung wet/dry weight ratio（W/D）， hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot were used to observe and detect the pathological changes of lung tissue， expression levels of inflammatory cytokine tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）， and expressions of TRPV1 and TRPA1 proteins in heat-sensitive channel， nuclear factor kappa-B （NF-κB）， inhibitor of NF-κB （IκBα） in inflammatory pathway， and phosphorylated proteins. The phosphorylated protein/total protein ratio was calculated. ResultCompared with that in the normal group， the lung tissue of mice in the model group was seriously damaged， and pulmonary capillary permeability increased. Alveolar capillary congestion and dilation destroyed the complete structure of the alveolar， and the alveolar wall thickened. A large number of inflammatory cells and red blood cells were infiltrated， and pulmonary edema was significantly aggravated. The expressions of TNF-α， IL-6， TRPV1， TRPA1， phosphorylated NF-κB p65/NF-κB p65， and phosphorylated IκBα/IκBα were significantly increased （P<0.01）， and the whole lung W/D was significantly increased （P<0.01）. Compared with the model group， the dexamethasone group and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction could significantly improve pulmonary edema. TNF-α， IL-6， TRPV1， TRPA1， lung tissue NF-κB p65， and IκBα phosphorylated protein/total protein ratio decreased significantly （P<0.05， P<0.01）. The whole lung W/D also decreased significantly （P<0.05， P<0.01）. ConclusionQingfei Paidu decoction has anti-inflammatory and protective effects on LPS-ALI mice， which can effectively reduce inflammation， induce diuresis， and alleviate edema. Its mechanism may be related to the regulation of the expression of TRPA1 and TRPV1 and the inhibition of the activation of the NF-κB pathway.

Objective:To investigate the feasibility of developing a radiomics model based on MRI and clinical features to predict the PD-L1 level in breast cancer.Methods:A total of 139 consecutive patients with breast cancer confirmed by pathology were enrolled retrospectively, including 79 PD-L1 negative patients and 60 PD-L1 positive patients. All patients were randomly assigned to a training dataset( n=97) and a validation dataset( n=42). Radiomics features were extracted from dynamic contrast-enhanced MRI. Radiomics feature selection was generated through the analysis of variance(ANOVA), least absolute shrinkage and selection operator(LASSO). Radiomics model and comprehensive model were developed for predicting the level of PD-L1. The receiver operating characteristic curve(ROC) was used to evaluate the predictive capacity of the models. Results:The radiomics model exhibited good performance in the training and validation datasets, with an area under the curve(AUC) of 0.847(95% confidence interval CI: 0.770-0.924) and 0.826(95% CI: 0.699-0.954), respectively. Compared with the radiomics model, the clinical feature combined prediction model showed better results, with AUC of 0.919(95% CI: 0.868-0.970) and 0.882(95% CI: 0.782-0.982), respectively, but without statistically significant difference( Z=1.32, P=0.19), respectively, but without statistically significant difference. Conclusions:The radiomi.Conclusions:The radiomics model has a certain value in preoperative prediction of PD-L1 expression level in breast cancer, which may be used as a supplement and improvement to the pathological gold standard to provide support for clinical decision-making.

OBJECTIVE: To compare two primary culture methods of human nasal epithelial (HNE) in vitro, and explore a suitable method to be used in further study. METHOD: Achievement ratio and growth curve of primarily cultural HNE by tissue piece culture were compared to those by isolated cell culture. Shape and appearance were observed and cellular sources were identified to get preliminary bionomics of HNE. RESULT: The isolated cell culture method (87.5%) was shown to be superior to tissue piece culture method (83.33%) by comparing achievement ratios, but no statistical significance was found (P > 0.05). The growth curve of isolated cell culture was higher than that of tissue piece culture. All cultured cells were confirmed coming from epithelial cells by observing shape, appearance and dyeing cytokeratin. CONCLUSION The isolated cell culture method is more suitable for primary culture due to its less promiscuity, faster proliferation, and more stable and reliable cell supply.
Cell Count ; Cell Culture Techniques ; methods ; Cells, Cultured ; Epithelial Cells ; cytology ; Humans ; Nasal Mucosa ; cytology

OBJECTIVE: To determine the effect of transforming growth factor-β1 (TGF-β1) on the expression of telomerase in hepatic stellate cells (HSCs) in rats and the role of TGF-β1 in the development of liver fibrosis. METHODS: Primary HSCs were isolated from normal rats by density gradient separation and divided into 2 groups for culturing. The morphology of HSCs was identified by the inverted fluorescence microscope. The purity of HSCs was identified by immunohistological expression and fluorescence analysis. One group of HSCs was treated with different concentrations (0, 0.1, 1, and 10 ng/mL) of TGF-β1 for 24 h, while the other group was treated with 1 ng/mL TGF-β1 and cultured for 3, 6, and 9 days. The mRNA expression of telomerase reverse transcriptase (TERT) was assessed and compared by polymerase chain reaction. RESULTS: Cell morphology showed that TGF-β1 triggered the differentiation of HSCs from a quiescent phenotype into highly activated myofibroblasts. TERT mRNA expression in the primary HSCs showed slight increase with the culture time, though with no statistical difference between the results at various time points (P>0.05). TGF-β1 at 0.1 ng/mL did not significantly affect the TERT mRNA level compared with the 0 ng/mL group, while 1 ng/mL and 10 ng/mL TGF-β1 significantly decreased the level of TERT mRNA (P<0.05). TGF-β1 at 1 ng/mL had only weak effect on TERT mRNA expression after the 3 day treatment compared with the 0 ng/mL group (P>0.05). TGF-β1 at 1 ng/mL significantly inhibited TERT mRNA expression 6 days after the treatment (P<0.05). TGF-β1 inhibited the expression of TERT mRNA level in the HSCs in both dose- and time-dependent manner. CONCLUSION TGF-β1 may contribute to the transdifferentiation of HSCs by reducing TERT levels to develop hepatic fibrosis.
Animals ; Cell Transdifferentiation ; Cells, Cultured ; Hepatic Stellate Cells ; drug effects ; metabolism ; RNA, Messenger ; Rats ; Telomerase ; metabolism ; Transforming Growth Factor beta1 ; pharmacology

Fibromyalgia syndrome (FMS), also known as fibromyalgia, is clinically characterized by diffuse chronic muscle and bone pain, accompanied by fatigue, sleep disturbances, depressive episodes, and cognitive and intestinal dysfunction. Due to lack of clear specific laboratory indicators and appropriate imaging examinations, FMS diagnosis is mostly based on clinical symptoms, but FMS clinical symptoms of lack specificity, and current clinical diagnostic criteria are mostly exclusive criteria, which is prone to missed diagnosis and misdiagnosis. In order to further promote the standardized diagnosis and treatment of FMS, this paper makes extensive references to laboratory-related diagnostic indexes of FMS (Tau, adiponectin, serum cathepsin S, cystatin C, serum ferritin, nitric oxide, neutrophil/lymphocyte ratio, platelet distribution width and mean platelet volume) at home and abroad, aiming to provide new ideas for early diagnosis and intervention of FMS.

Osteoporosis （OP） is a common bone disease affecting the quality of life and causing huge medical burden to the patients and society. The occurrence of OP is mainly caused by excessive bone resorption and insufficient bone formation， which are directly influenced by external calcium ion balance. Calcium imbalance can impair bone integrity， reduce the calcium supply to the bone， and lower the calcium content in the bone， thus triggering OP. Drugs are the main anti-OP therapy in modern medicine， which， however， may cause adverse reactions and drug dependence. Chinese medicines have good clinical effects and high safety in treating OP， being suitable for long-term use. Recent studies have shown that Chinese medicines can alleviate estrogen deficiency， regulate bone cell and calcium metabolism， which is crucial for the formation and development of OP. The transient receptor potential cation channel superfamily V members 5 and 6 （TRPV5 and TRPV6， respectively） affect bone homeostasis by mediating the transmembrane calcium ion transport in the intestine （TRPV6） and kidney （TRPV5）. Therefore， TRPV5/6 is one of the key targets to understand the anti-OP mechanisms of the effective parts of Chinese medicines， which is worthy of further study. This paper summarizes the research results about the anti-OP effects of Chinese medicines in the last two decades， especially the mechanism of regulating calcium metabolism， aiming to provide new ideas for the basic research， clinical application， and drug development of OP treatment.

ObjectiveTo investigate the mechanism of Renshentang， recorded in Synopsis of Golden Chamber， in the treatment of atherosclerosis （AS） based on the autophagic effect of transient receptor potential vanilloid subtype 1 （TRPV1） on arterial smooth muscle. MethodFourteen SPF-grade 8-week-old male C57BL/6J mice were assigned to the normal group and 70 8-week-old apolipoprotein E knockout （ApoE^-/-） mice were assigned to the experimental group. The AS model was induced by a high-fat diet in the mice in the experimental group for eight weeks. The model mice were then randomly divided into model group， low-， medium-， and high-dose Renshentang groups （2.715， 5.43， and 10.68 g·kg^-1·d^-1）， and simvastatin group （0.02 g·kg^-1·d^-1）. Drug treatment lasted eight weeks. Serum was taken and serum total cholesterol （CHO）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） levels were measured by assay kits to observe the changes in lipid levels in mice. The aorta was stained with hematoxylin-eosin （HE） to observe the overall pathology of the aortic root and oil red O staining was used to detect the lipid deposition in the aortic plaque and calculate the percentage of the aortic root area to the lumen area. The protein expression of TRPV1， adenylate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， autophagy effector-1 （Beclin-1）， and microtubule-associated protein 1 light chain 3 （LC3Ⅱ） in mouse aortic tissues was determined by Western blot. ResultCompared with the normal group， the model group showed increased serum CHO， TG， and LDL-C levels， decreased HDL-C， and increased aortic root plaque area （P<0.01）. Compared with the model group， the Renshentang groups showed decreased levels of CHO， TG， and LDL-C in serum （P<0.05， P<0.01）， especially in the low- and medium-dose Renshentang groups （P<0.01）. Compared with the normal group， the simvastatin group and the Renshentang groups showed reduced aortic root plaque area （P<0.05）， especially in the high-dose Renshentang group （P<0.01）. Compared with the normal group， the model group showed decreased relative expression levels of TRPV1， p-AMPK/AMPK， Beclin-1， and LC3Ⅱ/LC3Ⅰ（P<0.05， P<0.01）. Compared with the model group， the medium- and high-dose Renshentang groups showed increased relative expression levels of TRPV1， p-AMPK/AMPK， Beclin-1， and LC3Ⅱ/LC3Ⅰ（P<0.05，P<0.01）. ConclusionThe anti-AS effect of Renshentang recorded in Synopsis of Golden Chamber may be achieved by up-regulating TRPV1 expression to restore the level of autophagy mediated by AMPK.

ObjectiveTo explore the effect and mechanism of Zhishi Xiebai Guizhitang on the progression of atherosclerosis （AS） mice based on the regulation of cholesterol metabolism in foam cells by transient receptor potential channel ankyrin 1 （TRPA1）. MethodThe AS model was established on apolipoprotein E knockout （ApoE^-/-） mice with a high-fat diet. The mice were randomly divided into low-dose， middle-dose， and high-dose groups of Zhishi Xiebai Guizhitang （2.97， 5.94， 11.88 g·kg^-1） and simvastatin group （0.002 g·kg^-1）， and the drug was administered along with a high-fat diet. C57BL/6J mice were fed an ordinary diet as a normal group. After the above process， the aorta and serum of mice were taken. The pathological changes of the aortic root were observed by hematoxylin-eosin （HE） staining. The lipid plaques in the aorta were observed by gross oil redness. Serum levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein cholesterol （LDL-C）， and high density lipoprotein cholesterol （HDL-C） were detected， and the levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot and immunohistochemical method were used to analyze the expression of TRPA1， ATP-binding cassette transporter A1 （ABCA1）， ATP-binding cassette transporter G1 （ABCG1）， and mannose receptor （CD206）. ResultFrom the perspective of drug efficacy， compared with the normal group， pathological changes such as plaque， a large number of foam cells， and cholesterol crystals appeared in the aorta of the model group， and the serum levels of TC， LDL-C， IL-1β， and IL-18 were significantly increased （P<0.01）. The HDL-C level was significantly decreased （P<0.01）， and the CD206 level in aortic tissue was significantly decreased （P<0.01）. Compared with the model group， the lipid deposition in the aorta was alleviated in all drug administration groups. In addition， except for the high-dose group of Zhishi Xiebai Guizhitang， all drug administration groups could significantly decrease the levels of TC and LDL-C （P<0.01）. In terms of inflammation， except for the middle-dose group of Zhishi Xiebai Guizhitang， the levels of IL-1β and IL-18 were significantly decreased in all drug administration groups （P<0.05）. Moreover， Zhishi Xiebai Guizhitang could also up-regulate the levels of CD206， and the difference was significant in the middle-dose and high-dose groups （P<0.05）. From the perspective of mechanism， the expression levels of TRPA1， ABCA1， and ABCG1 in the aorta in the model group were lower than those in the normal group （P<0.05）. Compared with the model group， all drug administration groups significantly increased the expression of TRPA1 in the aorta （P<0.05）， and the expressions of ABCA1 and ABCG1 were increased. The differences in the middle-dose and high-dose groups and the simvastatin group were significant （P<0.05）， which was basically consistent with the trend of immunohistochemical results. ConclusionZhishi Xiebai Guizhitang can effectively reduce blood lipid and inflammation levels and inhibit the formation of aortic plaque. The mechanism may be explained as follows： the expressions of ABCA1 and ABCG1 downstream are increased through TRPA1， which promotes cholesterol outflow in foam cells， thereby regulating cholesterol metabolism， intervening in inflammation level to a certain extent， and finally treating AS.