ObjectiveTo investigate the effect of astragaloside Ⅳ（AS-Ⅳ） on the proliferation of vascular smooth muscle cells（VSMCs） induced by angiotensin Ⅱ（Ang Ⅱ） based on solute carrier family 7 member 11/glutathione peroxidase 4（SLC7A11/GPX4） pathway. MethodsPrimary rat thoracic aortic VSMCs were cultured by tissue explant method， and the cell types were identified by immunofluorescence. Cell counting kit-8（CCK-8） was used to determine the optimal concentration and time of AS-Ⅳ after Ang Ⅱ stimulation. The experiment was divided into blank group， model group， AS-Ⅳ group（40 μmol·L^-1）， Erastin group（0.5 μmol·L^-1）， Erastin+AS-Ⅳ group（0.5 μmol·L^-1+40 μmol·L^-1）. The blank group was cultured in normal medium， the model group was cultured in medium containing Ang Ⅱ（0.1 μmol·L^-1）， and each administration group was cultured in medium containing Ang Ⅱ（0.1 μmol·L^-1） and the corresponding doses of drug. CCK-8 and plate clone formation assay were used to detect the proliferation of cells in each group， Prussian blue staining was used to detect cell iron deposition， the content of reactive oxygen species（ROS） in cells was detected by fluorescence probe method， the content of malondialdehyde（MDA） was detected by thiobarbituric acid（TBA） method， and the protein levels of SLC7A11 and GPX4 in each group were detected by Western blot. ResultsPrimary rat thoracic aortic VSMCs were successfully cultured by tissue explant method， and immunofluorescence detection showed that positive expression of α-smooth muscle actin（α-SMA） and negative expression of vimentin in the cells， identifying them as VSMCs. The optimal concentration and time of AS-Ⅳ determined by CCK-8 were 40 μmol·L^-1 and 24 h， respectively. Pharmacodynamic studies showed that compared with the blank group， the cell proliferation in the model group increased， the iron deposition in the cells increased， the contents of ROS and MDA increased， and the expression levels of SLC7A11 and GPX4 proteins decreased（P<0.05， P<0.01）. Compared with the model group， the cell proliferation of the AS-Ⅳ group was inhibited， the iron deposition in the cells was decreased， the contents of ROS and MDA were decreased， and the expression levels of SLC7A11 and GPX4 proteins were increased（P<0.05， P<0.01）. While in the Erastin group， the cell proliferation was increased， the iron deposition was increased， ROS and MDA contents were increased， and the expression levels of SLC7A11 and GPX4 proteins were decreased（P<0.05， P<0.01）. Compared with the AS-Ⅳ group， Erastin+AS-Ⅳ group showed increased cell proliferation， increased iron deposition in cells， increased ROS and MDA contents， and decreased expression of SLC7A11 and GPX4 proteins（P<0.05）. Compared with the Erastin group， the cell proliferation in Erastin+AS-Ⅳ group was inhibited， the iron deposition was decreased， the contents of ROS and MDA were decreased， and the expression levels of SLC7A11 and GPX4 proteins were increased（P<0.05， P<0.01）. ConclusionAS-Ⅳ can inhibit ferroptosis by regulating the SLC7A11/GPX4 pathway， so as to weaken the proliferation of VSMCs， thus playing a role in the treatment of atherosclerosis.

ObjectiveTo identify the chemical components of Houpo Wenzhongtang in vivo and in vitro and to analyze the pharmacokinetic properties of the index components in rats with deficiency-cold of spleen and stomach. MethodThe chemical components of Houpo Wenzhongtang was analyzed and identified by ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS）. Six rats were randomly selected from 18 SD rats as the blank group， and the remaining rats were given lard and cold vinegar for a long time to construct a rat model with deficiency-cold of spleen and stomach. After successful modeling， the rats were randomly divided into the model group and Houpu Wenzhongtang group（13.5 g·kg^-1， calculated as crude drug）. The administration group was given the corresponding dose of Houpu Wenzhongtang by gavage， and the blank group and the model group were given the same amount of distilled water by gavage. Enzyme-linked immunosorbent assay（ELISA） were used to measure gastrin（GAS） and motilin（MTL） levels in each group. At the same time， plasma samples were collected at different time points after administration， and blood-entry prototype components and metabolites of Houpo Wenzhongtang were analyzed by UPLC-Q-TOF-MS/MS. On this basis， plasma concentrations of magnolol， honokiol， alpinetin and hesperidin in Houpo Wenzhongtang were determined by ultra performance liquid chromatography coupled with triple quadrupole/linear ion trap mass spectrometry（UPLC-QTRAP-MS/MS）， and the pharmacokinetic parameters were calculated using DAS 2.0 software. ResultA total of 79 chemical components， including 44 flavonoids and 11 lignans， were identified in Houpo Wenzhongtang. Meanwhile， 18 blood-entry prototype components and 27 metabolites were identified， the main metabolic pathways of metabolites were glucuronidation， sulfation， oxidation and hydrolysis， and phase Ⅰ and phase Ⅱ were the two primary forms of metabolism. Pharmacokinetic results showed that among the four index components， the time to peak（t_max） values of magnolol and honokiol were consistent and exhibited similar drug metabolism characteristics， the t_max of alpinetin was the shortest， and the absorption rate was the fastest， which had the earliest peak plasma concentration levels， and hesperidin had the shortest mean residence time（MRT_0-t） and the highest metabolic rate in rats. ConclusionThis study clarifies the blood-entry prototype components and their metabolites of Houpo Wenzhongtang in the rat model of deficiency-cold of spleen and stomach， and reveals the pharmacokinetic characteristics of the main active ingredients， which can provide a scientific basis for the study of pharmacodynamic material basis of this formula and its clinical application in treating the syndrome of deficiency-cold of spleen and stomach.

Objective:To explore the psychological experience and demand of parents of children with delayed recovery after congenital heart disease surgery.Methods:This study adopted phenomenological research methods from qualitative research. Using the purposive sampling method, parents of postoperative delayed recovery children with congenital heart disease who met the inclusion criteria were selected as the research objects from October to November 2019 at Fuwai Hospital, Chinese Academy of Medical Sciences. Semi-structured interviews were conducted with the parents of the children, and the data were analyzed by Colaizzi 7-step analysis method.Results:Finally, 13 parents of children with delayed recovery after congenital heart disease surgery were included. According to the interview results, four themes were extracted, which were negative psychological experience of parents of children with delayed recovery, positive psychological experience and expectation change of parents, heavy economic burden of parents and diversified needs of parents.Conclusions:During the delayed recovery period, psychological experience of parents is complex and their needs are diverse. The nursing staff should identify and pay attention to the causes of the negative psychological experience of the parents of the children, timely channel their negative emotions and strengthen the positive psychological experience in many aspects. They can assist parents to seek social help to reduce physical and mental pressure and meet the diverse needs of parents by providing high-quality nursing services and multi-channel information support.

Objective To observe the effects of methylenedioxygenase(TET2)expression on the proliferation ability,apoptosis rate,cell cycle,migration,and invasion of non-small cell lung cancer cells.Methods Detect the expression of TET2 gene and protein in different non-small cell lung cancer cells through RT-PCR and Western blot.Knock down the gene expression of the highest TET2 expressing cells(shRNA1)and overexpress the gene in the most low TET2 expressing cells(OE-TET2).Detect the effect of TET2 expression on cell proliferation through CCK-8,observe the effect of TET2 expression on cell apoptosis and cell cycle through flow cytometry,and observe the effect of TET2 expression on cell migration and invasion ability through migration and invasion experiments.Results The expression levels of TET2 gene and protein in H1299 cells were the highest,while A549 cells had the lowest expression levels(P＜0.05).The proliferation ability of OE-TET2 group cells was significantly lower than that of normal A549,while the proliferation ability of shRNA1 group cells was significantly higher than that of normal H1299 cells(P＜0.05).The apoptosis rate of OE-TET2 group was significantly higher than that of normal A549 cells,while the apoptosis rate of shRNA1 group was significantly lower than that of normal H1299 cells(P＜0.05).The number of migration and invasion cells in the OE-TET2 group was significantly lower than that of normal A549 cells,while the migration and invasion cells in the shRNA1 group were significantly higher than those in normal H1299 cells(P＜0.05).Conclusion TET2 can reduce the proliferation ability,increase its apoptosis rate,and slow down its migration and invasion ability of non-small cell lung cancer cells.

Background: The purpose of this research was to assess the role of heparanase (HPSE)/syndecan1 (SDC1)erve growth factor (NGF) on cancer pain from melanoma. Methods: The influence of HPSE on the biological function of melanoma cells and cancer pain in a mouse model was evaluated. Immunohistochemical staining was used to analyze HPSE and SDC1. HPSE, NGF, and SDC1 were detected using western blot. Inflammatory factors were detected using ELISA assay. Results: HPSE promoted melanoma cell viability, proliferation, migration, invasion, and tumor growth, as well as cancer pain, while SST0001 treatment reversed the promoting effect of HPSE. HPSE up-regulated NGF, and NGF feedback promoted HPSE. High expression of NGF reversed the inhibitory effect of HPSE down-regulation on melanoma cell phenotype deterioration, including cell viability, proliferation, migration, and invasion. SST0001 down-regulated SDC1 expression. SDC1 reversed the inhibitory effect of SST0001 on cancer pain. Conclusions The results showed that HPSE promoted melanoma development and cancer pain by interacting with NGF/SDC1. It provides new insights to better understand the role of HPSE in melanoma and also provides a new direction for cancer pain treatment.

Objective:To investigate the distribution law of TCM syndrome types and the differences in influencing factors among different syndrome types in unstable angina pectoris (UA), so as to provide an objective basis for TCM syndrome differentiation and treatment of UA.Methods:A retrospective study chose 1 684 inpatients in the Department of Cardiovascular Medicine of the First Affiliated Hospital of Henan University of Chinese Medicine from August 2015 to April 2019. Epidata 3.0 software was used to input general information of patients [gender, age, length of hospital stay, BMI, systolic blood pressure (SBP), diastolic blood pressure (DBP)], laboratory indicators[TC, TG, HDL-C, LDL-C, fibrinogen (FIB), thrombin time (TT), D-dimer (D-dimer), HbA1c], TCM syndrome types (qi and yin deficiency syndrome, phlegm turbidity and obstruction syndrome, qi deficiency and blood stasis syndrome, heart blood stasis syndrome, qi stagnation and blood stasis syndrome, heart and kidney yang deficiency syndrome) and other information. A database was established to analyze the distribution of TCM syndrome types and the relationship with the general information of patients, blood lipids, blood sugar and coagulation indexes. Logistic regression was used to analyze the influencing factors of different syndrome types.Results:The distribution of syndrome types in 1 684 UA patients was as follows: qi and yin deficiency syndrome (44.7%), phlegm turbidity and obstruction syndrome (35.3%), qi deficiency and blood stasis syndrome (7.4%), heart blood stasis syndrome (5.3%), qi stagnation and blood stasis syndrome (4.6%), heart and kidney yang deficiency syndrome (2.6%); more men than women ( P<0.05); there were significant differences in the distribution of gender, age, BMI, TC, and HDL-C among the 6 syndrome types ( P<0.05); the age of patients with phlegm turbidity and obstruction syndrome was younger than that of qi and yin deficiency syndrome and heart blood stasis syndrome ( P<0.05); the age of patients with qi stagnation and blood stasis syndrome was younger than that of qi and yin deficiency syndrome, heart blood stasis syndrome, and heart kidney yang deficiency syndrome ( P<0.05); BMI of patients with phlegm turbidity and obstruction syndrome was higher than that of qi and yin deficiency syndrome and qi stagnation and blood stasis syndrome ( P<0.05); the level of TC in patients with phlegm turbidity and obstruction syndrome was lower than that of qi and yin deficiency syndrome and qi deficiency and blood stasis syndrome ( P<0.05); the level of HDL in patients with qi and yin deficiency syndrome was lower than that in qi deficiency and blood stasis syndrome and qi stagnation and blood stasis syndrome. Binary Logistic regression analysis found that TC [ OR(95% CI)=0.761(0.592, 0.978)] and HDL-C [ OR(95% CI)=2.131(1.145, 3.966)] were independent influencing factors for predicting qi deficiency and blood stasis syndrome ( P<0.05); age[ OR(95% CI)=1.017 (1.008, 1.026)], length of hospital stay [ OR(95% CI)=1.019 (1.001, 1.038)], DBP [ OR(95% CI)=0.984(0.975, 0.993)] and HDL-C [ OR(95% CI)=0.984(0.975, 0.993)] were independent influencing factors for predicting qi and yin deficiency syndrome ( P<0.05); age [ OR(95% CI)=0.965 (0.946, 0.985)], and HDL-C [ OR(95% CI)=2.329(1.206, 4.500)] were independent influencing factors for predicting qi stagnation and blood stasis syndrome ( P<0.05); age [ OR(95% CI)=0.982 (0.973, 0.991)], length of hospital stay [ OR(95% CI)= 0.978 (0.958, 0.997)], BMI [ OR(95% CI)=1.048 (1.015, 1.082)], DBP [ OR(95% CI)=1.014 (1.004, 1.024)] and HDL-C [ OR(95% CI)=0.505 (0.351, 0.726)] were independent influencing factors for predicting phlegm turbidity and obstruction syndrome ( P<0.05); age [ OR(95% CI)=1.031(1.003, 1.060)] and DBP [ OR(95% CI)=1.028(1.001, 1.056)] were independent influencing factors for predicting heart kidney yang deficiency syndrome ( P<0.05). Conclusion:The distribution of TCM syndrome types in UA shows a certain regularity, among which qi and yin deficiency syndrome and phlegm turbidity and obstruction syndrome are more common. Gender, age, BMI, TC, HDL-C are different among TCM syndrome types, which can provide some reference for UA TCM syndrome differentiation and treatment.

Objective:To explore whether the electroacupunture stimulation (ES) at acupoint Zusanli (ST36) can inhibit the bone loss caused by Staphylococcus aureus (SA) infection and its mechanism in a model of SA osteomyelitis.Methods:Twelve male C57 BL/6 mice aged 10 to 12 weeks were randomly divided into 2 even groups ( n=6) for SA infection + ES or SA infection only. After ES at ST36 was conducted for 4 weeks in the model of SA osteomyelitis, samples were harvested from the femora and tibiae. Micro-CT reconstruction was performed to detect trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), connectivity density (Conn.Dn) to analyze changes in bone mass. Leptin receptor (LEPR) staining was performed to detect osteoblasts. Tartrate resistant acid phosphatase (TRAP) staining was used to detect the changes in osteoclasts. The changes in plasma inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). Results:Micro-CT results showed that the BV/TV, Tb.N, Tb.Th, and Conn.Dn in the cancellous bone in the target areas in the SA + ES group were all higher than those in the SA group, LEPR immunofluorescence results indicated that the number of osteogenic precursor cells in the ES group was larger than that in the SA group, and serum ELISA indicated a decrease in inflammatory factors in the blood in the SA+ES group compared with the SA group. There were significant differences in the comparisons above ( P<0.05). There was no significant difference in the number of osteoclasts on the surface of trabecular bone between the 2 groups in TRAP staining. Conclusion:ES may slow down infectious bone destruction by inhibiting the inflammatory response induced by SA infection and by inducing aggregation and differentiation of mesenchymal stem cells into trabecular bone.

Objective:To study the change pattern of neck diameter and relevant factors in nasopharyngeal carcinoma (NPC) patients treated with helical tomotherapy, aiming to provide reference for clinical practice.Methods:Fifteen NPC patients treated with helical tomotherapy at Sun Yat-Sen University Cancer Center from November 2020 to February 2021 were enrolled in this study. The transverse diameters of NPC patients' neck contours and body weight of all patients during radiotherapy were recorded weekly. We used descriptive statistics to explore the distribution of transverse diameters of NPC patients' neck contours during radiotherapy. And Spearman correlation analysis was used to assess the association between weight loss and changes in neck contour transverse diameter.Results:During radiotherapy, the distribution of transverse diameters of NPC patients' neck contours was completely different from the initial values. Specifically, the transverse diameters were significantly reduced at the 4th week of the radiotherapy. Moreover, the reduction of transverse diameter of neck contour was highly correlated with the weight loss ( r=0.803, P<0.05), and moderately correlated with gender ( r=0.523, P<0.05). However, there was no statistical correlation between the alteration of neck diameter and age, TNM stage, and the mean dose of GTV nd-L, GTV nd-R, PTV nd-L and PTV nd-R (all P>0.05). Conclusions:The neck contours of NPC patients are altered regularly during helical tomotherapy, which are narrowed the most obviously in the 4th week. Extensive attention should be paid to the changes of neck contour during radiotherapy in clinical practice.

Objective:To introduce the design, manufacture and clinical application of the custom-made temporomandibular joint (TMJ)-skull base combined prosthesis and evaluate its safety, effectiveness and accuracy.Methods:The patients diagnosed with the TMJ-skull base lesion in Department of Oral Surgery, Shanghai Ninth People′s Hospital from October 2016 to November 2020 were recruited in this study. The maxillofacial CT data for all the patients were obtained and transformed into the Mimics 18.0 software preoperatively. The custom-made TMJ-skull base combined prosthesis, included four components, was designed based on the anatomy, stress distribution and movement of the TMJ and skull base, and fabricated by three-dimensional printing and 5-axis milling technologies. The TMJ-skull base lesion was excised completely with the help of digital templates from modified preauricular and/or post and submandibular incisions. The combined prosthesis were implanted and fixed after the lesion resection. The examinations including general situation, cranio-maxillofacial structure and function were taken during and after surgery to assess its using effect.Results:Ten patients [6 females and 4 males, (43.2±13.6) years old] were included and all prostheses were positioned accurately and fixed excellently. After (29.4±17.3) months follow-up, the occlusion relationship was stable and no adverse symptoms such as dizziness, headache, meningeal irritation and permanent facial nerve injury occurred. The pain, diet, mandibular movement function, lateral movement to diseased side and mouth opening had significant improvements. The forward movement and lateral movement to normal side were not improved significantly. There were no prosthesis displacement, loosening and fracture in X-ray and CT postoperatively. With the pre and postoperative craniomaxillofacial model merging, the maximal implanted error was (0.52±0.17) mm for fossa and condyle and (1.62±0.26) mm for skull base and mandibular handle in surface deviation analysis.Conclusions:The custom-made TMJ-skull base combined prosthesis with customized design and 3D printing fabrication is safe, effective and precise in clinical application.

Objective:To establish a classification model for rapid identification of hypervirulent subtype ST17 clones of Group B Streptococcus (GBS) using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS).Methods:In a retrospective study, 235 strains of GBS strains were selected from multiple centers in China during 2015-2018. For model generation,45 strains of ST17 and 50 strains of non-ST17 (20 ST19, 15 ST12 and 15 ST10 strains) were enrolled as the modeling group. The remaining 90 main ST strains (40 ST17, 16 ST10, 17 ST12 and 17 ST19) were served as validation group. 50 GBS strains classified as other minor ST subtypes were regarded as taxonomic groups. MS spectra were collected by Bruker mass spectrometry, and then loaded for model generation and verification, and screening of differential peptide peaks by genetic algorithm (GA) and model verification on ClinProTools 3.0 software.Results:The recognition rate for ST17-GA model were 99.4% with cross validation value of 96.9%. Among the ten differential peptide peaks for the classification model, the weights of both two main peptide peaks m/z 2 956 and m/z 5 912 were greater than 1, while the weights of the all other eight peptide peaks were less than 0.5. Model validation showed only one of the ST17 was misjudged as non-ST17 strain, resulting in diagnostic accuracy of 98.9%, sensitivity of 97.5% and specificity of 100%, positive predictive value of 100% and negative predictive value of 98.0%, respectively. For other sporadic STs, 42.0% (21/50) of them were misdiagnosed as ST17 subtype.Conclusion:A MALDI-TOF MS classification model for hypervirulent subtype of ST17 GBS strains has been successfully established with good diagnostic efficacy.