Objective: To establish the content determination method of tanshinoneⅡ A in changshangfangban ointment. Methods: HPLC was performed to determine tanshinone Ⅱ A on ODS column, using methanol water(79∶21,v∶v) as a mobile phase and detection wavelength at 268nm. Results: The linearity of this method was well, and the average recovery was 98.9%,RSD=0.7%. Conclusion: This method was simple, quick and accurate, and available for quality control of chuangshangfangban ointment.

Objective To investigate the expression level of tetratricopeptide repeat protein 9A in tumors and its association with the patients'prognosis and immune infiltration.Methods TTC9A expression in different tumor tissues and its association with prognosis,DNA methylation,tumor mutation burden(TMB),and microsatellite instability(MSI)were analyzed based on data from TCGA and GTEx.TIMER and xCell were used to analyze the relationship between TTC9A expression and immune infiltration.Western blotting and RT-qPCR were used to detect the expression of TTC9A in 4 types of cancer cell lines.Results TTC9A expressions were significantly increased in many tumors and down-regulated in a few cancer types(P<0.05).Western blotting and RT-qPCR showed that TTC9A expressions were elevated in lung,colon and liver cancer cells but decreased in bladder cancer cells.In head and neck squamous cell carcinoma,renal clear cell carcinoma,renal papillary cell carcinoma,low-grade glioma,malignant mesothelioma,and endometrial carcinoma tumors,a high expression of TTC9A was strongly correlated with better overall survival(OS),disease-specific survival(DSS),and progression-free interval(PFI)(P<0.05),but was correlated with worse OS,DSS,and PFI in lung adenocarcinoma,pancreatic adenocarcinoma,adrenal carcinoma,and rectal adenocarcinoma(P<0.05).TTC9A hypermethylation was associated with a more favorable prognosis of glioblastoma multiforme,low-grade glioma,uveal melanoma,and ovarian plasmacytoid cystadenocarcinoma(P<0.05)but with poor prognosis of squamous cell carcinoma of the uterine cervix and intracervical adenocarcinoma,squamous cell carcinoma of head and neck,squamous cell carcinoma of the lungs,adrenal carcinoma,and endometrial carcinoma(P<0.05).In most of the cancer types,TTC9A was significantly correlated with the level of immune cell infiltration(P<0.05).Conclusion TTC9A can be used as a prognostic marker for a variety of cancers and is strongly associated with TBM,MSI and immune cell infiltration.

Objective To investigate the expression level of tetratricopeptide repeat protein 9A in tumors and its association with the patients'prognosis and immune infiltration.Methods TTC9A expression in different tumor tissues and its association with prognosis,DNA methylation,tumor mutation burden(TMB),and microsatellite instability(MSI)were analyzed based on data from TCGA and GTEx.TIMER and xCell were used to analyze the relationship between TTC9A expression and immune infiltration.Western blotting and RT-qPCR were used to detect the expression of TTC9A in 4 types of cancer cell lines.Results TTC9A expressions were significantly increased in many tumors and down-regulated in a few cancer types(P<0.05).Western blotting and RT-qPCR showed that TTC9A expressions were elevated in lung,colon and liver cancer cells but decreased in bladder cancer cells.In head and neck squamous cell carcinoma,renal clear cell carcinoma,renal papillary cell carcinoma,low-grade glioma,malignant mesothelioma,and endometrial carcinoma tumors,a high expression of TTC9A was strongly correlated with better overall survival(OS),disease-specific survival(DSS),and progression-free interval(PFI)(P<0.05),but was correlated with worse OS,DSS,and PFI in lung adenocarcinoma,pancreatic adenocarcinoma,adrenal carcinoma,and rectal adenocarcinoma(P<0.05).TTC9A hypermethylation was associated with a more favorable prognosis of glioblastoma multiforme,low-grade glioma,uveal melanoma,and ovarian plasmacytoid cystadenocarcinoma(P<0.05)but with poor prognosis of squamous cell carcinoma of the uterine cervix and intracervical adenocarcinoma,squamous cell carcinoma of head and neck,squamous cell carcinoma of the lungs,adrenal carcinoma,and endometrial carcinoma(P<0.05).In most of the cancer types,TTC9A was significantly correlated with the level of immune cell infiltration(P<0.05).Conclusion TTC9A can be used as a prognostic marker for a variety of cancers and is strongly associated with TBM,MSI and immune cell infiltration.

AIM:To investigate the role and molecular mechanisms of SMARCA4(SWI/SNF-related,matrix-associated,actin-dependent regulator of chromatin,subfamily A,member 4)in ferroptosis.METHODS:(1)Human fi-brosarcoma HT1080 cells were treated with dimethyl sulfoxide(DMSO)and different concentrations(31.25,62.5 and 125 nmol/L)of Ras-selective lethal small molecule 3(RSL3;ferroptosis inducer).Each treatment had 3 replicate wells of cells.The protein levels of SMARCA4 were detected by Western blot.(2)Two small interfering RNAs(siSMARCA4-1 and siSMARCA4-2)were constructed according to the SMARCA4 gene sequence.After SMARCA4 knockdown,each treat-ment had 3 replicate wells of cells,and the protein levels of SMARCA4 were determined by Western blot.Effects of DMSO,necrostatin 2 racemate(Nec-1s;necroptosis inhibitor),Z-VAD(OMe)-FMK(Z-VAD,pan-caspase inhibitor/apoptosis inhibitor)and ferrostatin-1(Fer-1,ferroptosis inhibitor)on cell viability were assessed using high-content analy-sis.The levels of ferroptosis indicators,including prostaglandin-endoperoxide synthase 2(PTGS2)transcription,lipid peroxidation,reactive oxygen species(ROS),labile iron pool(LIP)and glutathione,were determined by RT-qPCR and flow cytometry.The mRNA expression levels of pivotal iron metabolism genes,ferroptosis-related ROS regulatory genes,and cholesterol synthesis-related genes were measured using RT-qPCR.Impact of cholesterol on the cell viability were as-sessed using high-content analysis.(3)Common differential gene analysis and gene ontology(GO)enrichment analysis were performed on published online data.RESULTS:(1)Treatment with RSL3 significantly reduced the protein level of SMARCA4(P＜0.05).(2)Knockdown of SMARCA4 resulted in ferroptosis.(3)Knockdown of SMARCA4 did not induce ferroptosis by modulating the LIP and the transcription levels of ROS-related genes.(4)Knockdown of SMARCA4 affected the pathways associated with the cell membrane,lipid raft,and cholesterol synthesis.(5)Addition of cholesterol to cell culture medium rescued the ferroptosis induced by SMARCA4 knockdown(P＜0.01).CONCLUSION:Treatment with RSL3 reduces the protein level of SMARCA4 in human fibrosarcoma HT1080 cells,and inhibition of cholesterol synthesis by SMARCA4 knockdown leads to the ferroptosis of HT1080 cells.

OBJECTIVE To discuss the factors affecting the blood concentration of high-dose methotrexate （HD-MTX） and the occurrence of adverse drug reactions （ADR） when treating lymphoma with HD-MTX. METHODS From July 2020 to November 2021， the information of HD-MTX patients who had been monitored for HD-MTX blood drug concentration in the First Affiliated Hospital of Guangdong Pharmaceutical University was collected by retrospective analysis， such as medical record number， age， sex， height， body mass， chemotherapy plan， dosage； test indexes such as alanine transaminase， aspartate transaminase， total bilirubin， creatinine clearance （CrCl）， albumin （ALB） and other indexes were also collected before and after administration. The blood concentrations （c6 h， c24 h， c48 h） of HD-MTX were recorded， drug information of proton pump inhibitors （PPIs） was extracted and used， and ADR occurring within 48 h after administration were all evaluated. Single factor analysis， multiple linear regression and χ2 test were used to analyze the influential factors. RESULTS A total of 133 patients were included in this paper. The results of the single factor analysis of HD-MTX blood drug concentration showed that age， CrCl had an effect on c 6 h （P＜0.05）； age， CrCl and ALB had an effect on c24 h （P＜0.05）； age， body mass index （BMI）， CrCl， combined use of PPIs and ALB had an effect on c48 h （P＜0.05）. The results of multiple linear regression analysis showed that age and CrCl had no effect on c 6 h （P＞0.05）， age was the main influential factor of c 24 h （P＜0.05）， and CrCl and combined use of PPIs were the main influential factors of c48 h （P＜0.05）； the coefficient of variance expansion was between 1 and 3.5， indicating that the analysis results were acceptable. The overall incidence of adverse reactions was 51.13%， of which the blood and lymphatic system reactions were the most common. The results of the influential factors of ADR showed that age， BMI， liver function and CrCl had effect on the incidence of ADR （P＜0.05）. CONCLUSIONS During the process of HD-MTX in the treatment of lymphoma， the patient’s age， CrCl and combined use of PPIs should be considered， and the patient’s blood concentration should be monitored； at the same time， the age of patients， BMI， liver function and CrCl have an impact on the incidence of ADR.

Raman imaging yields high specificity and sensitivity when compared to other imaging modalities, mainly due to its fingerprint signature. However, intrinsic Raman signals are weak, thus limiting medical applications of Raman imaging. By adsorbing Raman molecules onto specific nanostructures such as noble metals, Raman signals can be significantly enhanced, termed surface-enhanced Raman scattering (SERS). Recent years have witnessed great interest in the development of SERS nanoprobes for Raman imaging. Rationally designed SERS nanoprobes have greatly enhanced Raman signals by several orders of magnitude, thus showing great potential for biomedical applications. In this review we elaborate on recent progress in design strategies with emphasis on material properties, modifying factors, and structural parameters.

OBJECTIVES: To evaluate the response to cardiac resynchronization therapy (CRT) and the correlation between CRT and pulmonary artery hemodynamic parameters. METHODS: The patients with chronic heart failure indicator for CRT were enrolled. The left ventricular end-systolic volume (LVESV) was measured by echocardiography and New York Heart Association (NYHA) classification was evaluated between one week before and six months after CRT. Mean pulmonary artery pressure (mPAP), pulmonary artery systolic pressure (PASP) and pulmonary vascular resistance (PVR) were measured by right heart catheterization. Left ventricular reverse remodeling (LVRR) is defined as a decrease of 15% or more in LVESV at the 6th month after CRT; Clinical response is defined as a decrease of NYHA classification at or above grade 1 at the 6th month after CRT. Pulmonary hypertension (PH) was defined as mPAP≥25 mmHg. According to the response, patients were divided into 3 groups: group A (LVRR+clinical response), group B (no LVRR+clinical response) and group C (no LVRR+no clinical response). The changes of NYHA classification, echocardiographic and pulmonary hemodynamic parameters were observed in the 3 groups. The Kaplan-Meier survival curve was used to analyze the differences in all-cause mortality, combined end-point events of death or re-hospitalization due to heart failure among different groups. RESULTS: A total of 45 patients with CRT implantation [aged (63.27±9.55) years, 36 males] were included. The average follow-up period was (33.76±11.50) months. Thirty-one patients (68.89%) were in group A, 9 of whom with PH. Eight patients (17.78%) were in group B, 7 of whom with PH. Six patients were in group C, all with PH. Cardiac function including NYHA classification, echocardiographic and pulmonary hemodynamic parameters had been significantly improved in group A after CRT implantation (<0.05). In group B, NYHA classification and pulmonary hemodynamic parameters were decreased significantly (<0.05), but echocardiographic parameters did not change obviously (>0.05). There were no significant changes in NYHA classification, echocardiographic and pulmonary hemodynamic parameters in group C (>0.05). Compared with group C, group A and group B had lower all-cause mortality (=0.005) and lower incidence of composite endpoint events (=0.001). CONCLUSIONS Patients with LVRR and clinical response after CRT have a good prognosis. Patients with clinical response but without LVRR have a better prognosis than those without clinical response and LVRR, which may be related to the decrease of pulmonary hemodynamic parameters such as mPAP and TPG.
Aged ; Cardiac Resynchronization Therapy ; Heart Failure ; therapy ; Hemodynamics ; Humans ; Male ; Middle Aged ; Pulmonary Artery ; Treatment Outcome ; Ventricular Remodeling

Objective    To characterize surgical outcomes of supra-cardiac total anomalous pulmonary venous connection (TAPVC), investigate risk factors for postoperative death, and explore informed LASSO machine learning methods to solve "small sample size problem" in research of rare congenital heart diseases. Methods    A retrospective analysis of 241 patients with supra-cardiac TAPVC who underwent surgical repair in Guangdong Provincial People's Hospital from 2009 to 2019 was conducted, including 179 males and 62 females with a median surgical age of 71 (33, 232) d. Detailed clinical data of the postoperative death-related factors were extracted. Univariable Cox proportional hazard models were used to initially screen potential risk factors for postoperative death. Factors with P鈮?.05 were retained. To solve the limitation of small sample size and the "P>n" problem, we proposed a novel LASSO method for conducting multivariable Cox regression analysis that was capable of bringing in findings of related studies to improve analysis power and to reduce false-negative findings. Results 聽 聽Univariable Cox analyses showed several potential clinical risk factors, among which highly significant factors (P<0.001) included: surgical weight鈮?.5 kg (HR=16.00), main pulmonary artery diameter (HR=0.78), prolonged cardiopulmonary bypass time (HR=1.21), aortic block time (HR=1.28), and postoperative ventilator-assisted time (HR=1.13/d). LASSO multivariable analysis revealed that independent risk factors for postoperative death included cardiopulmonary bypass time (aHR=1.308/30 min), age (aHR=0.898), postoperative ventilator-assisted time (aHR=1.023/d), weight鈮?.5 kg (aHR=2.545), right vertical venous return (aHR=1.977), preoperative pulmonary venous obstruction (aHR=1.633) and emergency surgery (aHR=1.383). Conclusion 聽 聽Our proposed informed LASSO method can use previous studies' results to improve the power of analysis and effectively solve the "P>n" and small sample size limitation. Cardiopulmonary bypass time, surgical age, postoperative ventilator-assisted time, weight, right vertical venous return, preoperative pulmonary venous obstruction, and emergency surgery are risk factors for postoperative death of supra-cardiac TAPVC.

BACKGROUND: Gallbladder cancer (GBC) is the most common malignant tumor of biliary tract. Isoliquiritigenin (ISL) is a natural compound with chalcone structure extracted from the roots of licorice and other plants. Relevant studies have shown that ISL has a strong anti-tumor ability in various types of tumors. However, the research of ISL against GBC has not been reported, which needs to be further investigated. METHODS: The effects of ISL against GBC cells in vitro and in vivo were characterized by cytotoxicity test, RNA-sequencing, quantitative real-time polymerase chain reaction, reactive oxygen species (ROS) detection, lipid peroxidation detection, ferrous ion detection, glutathione disulphide/glutathione (GSSG/GSH) detection, lentivirus transfection, nude mice tumorigenesis experiment and immunohistochemistry. RESULTS: ISL significantly inhibited the proliferation of GBC cells in vitro . The results of transcriptome sequencing and bioinformatics analysis showed that ferroptosis was the main pathway of ISL inhibiting the proliferation of GBC, and HMOX1 and GPX4 were the key molecules of ISL-induced ferroptosis. Knockdown of HMOX1 or overexpression of GPX4 can reduce the sensitivity of GBC cells to ISL-induced ferroptosis and significantly restore the viability of GBC cells. Moreover, ISL significantly reversed the iron content, ROS level, lipid peroxidation level and GSSG/GSH ratio of GBC cells. Finally, ISL significantly inhibited the growth of GBC in vivo and regulated the ferroptosis of GBC by mediating HMOX1 and GPX4 . CONCLUSION ISL induced ferroptosis in GBC mainly by activating p62-Keap1-Nrf2-HMOX1 signaling pathway and down-regulating GPX4 in vitro and in vivo . This evidence may provide a new direction for the treatment of GBC.
Animals ; Mice ; Carcinoma in Situ ; Chalcones/pharmacology* ; Ferroptosis ; Gallbladder Neoplasms/genetics* ; Glutathione Disulfide ; Kelch-Like ECH-Associated Protein 1 ; Mice, Nude ; NF-E2-Related Factor 2/genetics* ; Reactive Oxygen Species ; Humans