OBJECTIVE: Hepatocellular carcinoma (HCC) is a leading cause of mortality worldwide. Huachansu (Cinobufacini) is active extract isolated from the dry skin of Bufo Bufo gargarizans. It has now been widely used in clinical treatment of cancer, this study is to clarify the material basis of down-regulation of thymidylate synthase (TYMS) induced by Huachansu. METHODS: Our study utilized UPLC-MS/MS to identify major bioactive components from Huachansu. Cell Counting Kit 8 (CCK-8) assay and clone formation assay were used to examine the cell viability of tumor cells. TYMS and γ-H2AX level were detected by using quantitative real-time RT-PCR and/or western blotting. Small interfering RNA (siRNA) transfection was used to explore whether inhibition of TYMS could enhance the suppressive effect of Huachansu on cell growth of HCC cells. RESULTS: In our study, firstly, we identify 21 major bioactive components from Huachansu. CCK-8 assay results showed that Huachansu and its bioactive bufadienolides (Bufalin, Bufotalin, Cinobufotalin, Desacetylcinobufagin, Arenobufagin, Telocinobufagin, and Resibufogenin) significantly inhibited the proliferation of HepG2 and SK-HEP-1 cells in a dose- and time-dependent manner. Further molecular mechanistic investigation demonstrates that Huachansu significantly suppresses thymidylate synthase (TYMS), the enzyme which provides the sole de novo source of thymidylate for DNA synthesis. The inhibition of TYMS could lead to cell-cycle block and DNA damage of HCC cells. Furthermore, we identified that Huachansu markedly increased γ-H2AX expression, which indicated the presence of DNA damage. Moreover, we confirmed that transfection of cells with small interfering RNA specific to TYMS could increase the suppressive effects of Huachansu on the HCC cells proliferation. Quantitative RT-PCR analysis showed that Huachansu treatment had no effect on the transcription level of TYMS. Furthermore, proteasomal inhibitor MG132 could block TYMS inhibition induced by Huachansu, and concomitant administration of protein synthesis inhibitor cycloheximide (CHX) with Huachansu could further suppress the protein level of TYMS, indicating that Huachansu promotes proteasome-dependent degradation of TYMS in liver cancer cells. More importantly, the bioactive bufadienolides of Huachansu such as Bufalin, Bufotalin, Cinobufotalin, Desacetylcinobufagin, Arenobufagin, Telocinobufagin, and Resibufogenin could also significantly restrain the protein level of TYMS, revealing the material basis of inhibition of TYMS exposed to Huachansu. 5-Fluorouracil (5-FU) is a TYMS inhibitor, we also evaluate the effects of the combined treatment of Huachansu with 5-FU, the results show that interactions between Huachansu and 5-FU are synergistic or antagonistic. Thus, in clinical, attention should be paid to the dosage of Huachansu in combination with 5-FU. CONCLUSION Huachansu inhibits the growth and induces DNA damage of human HCC cells through proteasome-dependent degradation of TYMS, bioactive bufadienolides are the material basis of down-regulation of TYMS induced by Huachansu.

Objective To construct a machine learning prediction model for postoperative liver injury in patients with non-liver surgery based on preoperative and intraoperative medication indicators.Methods A case-control study was conducted on 315 patients with liver injury after non-liver surgery selected from the databases developed by 3 large general hospitals from January 2014 to September 2022.With the positive/negative ratio of 1 ∶3,928 cases in corresponding period with non-liver surgery and without liver injury were randomly matched as negative control cases.These 1243 patients were randomly divided into the modeling group(n=869)and the validation group(n=374)in a ratio of 7∶3 using the R language setting code.Preoperative clinical indicators(basic information,medical history,relevant scale score,surgical information and results of laboratory tests)and intraoperative medication were used to construct the prediction model for liver injury after non-liver surgery based on 4 machine learning algorithms,k-nearest neighbor(KNN),support vector machine linear(SVM),logic regression(LR)and extreme gradient boosting(XGBoost).In the validation group,receiver operating characteristic(ROC)curve,precision-recall curve(P-R),decision curve analysis(DCA)curve,Kappa value,sensitivity,specificity,Brier score,and F1 score were applied to evaluate the efficacy of model.Results The model established by 4 machine learning algorithms to predict postoperative liver injury after non-liver surgery was optimal using the XGBoost algorithm.The area under the receiver operating characteristic curve(AUROC)was 0.916(95%CI:0.883～0.949),area under the precision-recall curve(AUPRC)was 0.841,Brier score was 0.097,and sensitivity and specificity was 78.95%and 87.10%,respectively.Conclusion The postoperative liver injury prediction model for non-liver surgery based on the XGBoost algorithm has effective prediction for the occurrence of postoperative liver injury.

Objective To explore the reliability and safety of continuous monitoring of vital signs in patients using wireless wearable monitoring devices after video-assisted thoracoscopic surgery (VATS) for lung cancer. Methods The patients undergoing VATS for lung cancer in West China Hospital, Sichuan University from May to August 2023 were prospectively enrolled. Both wireless wearable and traditional wired devices were used to monitor the vital signs of patients after surgery. Spearman correlation analysis, paired sample t test and ratio Bland-Altman method were used to test the correlation, difference and consistency of monitoring data measured by the two devices. The effective monitoring rate of the wireless wearable device within 12 hours was calculated to test the reliability of its continuous monitoring. Results A total of 20 patients were enrolled, including 15 females and 5 males with an average age of 46.20±11.52 years. Data collected by the two monitoring devices were significantly correlated (P<0.001). Respiratory rate and blood oxygen saturation data collected by the two devices showed no statistical difference (P>0.05), while heart rate measured by wireless wearable device was slightly lower (=−0.307±1.073, P<0.001), and the blood pressure (=1.259±5.354, P<0.001) and body temperature(=0.115±0.231, P<0.001) were slightly higher. The mean ratios of heart rate, respiratory rate, blood oxygen saturation, blood pressure and body temperature collected by the two devices were 0.996, 1.004, 1.000, 1.014, and 1.003, respectively. The 95% limits of agreement (LoA) and 95% confidence interval of 95%LoA of each indicator were within the clinically acceptable limit. The effective monitoring rate of each vital signs within 12 hours was above 98%. Conclusion The wireless wearable device has a high accuracy and reliability for continuous monitoring vital signs of patients after VATS for lung cancer, which provides a security guarantee for subsequent large-scale clinical application and further research.

This report describes a case of maturity-onset diabetes of the young(MODY)type 3(MODY3)complicated with type 5(MODY5),including the patient's clinical features,diagnosis,and treatment,and reviews relevant literature.Using next-generation sequencing of MODY(types 1-14)gene exons and Sanger sequencing for verification,the patient and her mother were assessed.Based on the clinical phenotype and genetic test results,the patient was diagnosed as MODY3 combined with MODY5.Treatment included insulin and linagliptin,with monitoring of blood glucose changes.Clinicians should enhance their understanding of MODY clinical phenotypes.In adolescents with diabetes who have congenital pancreatic and renal developmental defects,elevated high-density lipoprotein cholesterol,no spontaneous ketosis,insulin secretion defects,negative pancreatic autoantibodies,no significant insulin resistance,and who are not obese,gene testing should be conducted to screen for MODY.Accurate diagnosis and personalized treatment can aid in achieving glycemic control,improving quality of life,and optimizing reproductive planning.

Hypertensive intracerebral hemorrhage (ICH) is mostly single in basal ganglia, thalamus and pons. Simultaneous hemorrhage in other brain regions is relatively rare, accounting for only 5.6% of all hemorrhagic strokes, while bilateral symmetrical hemorrhage is extremely rare. A case of bilateral basal ganglia symmetrical hemorrhage is reported for clinical reference.

Objective:To evaluate the performance of the automated digital cell morphology instrument in detecting platelet (PLT) clumps.Methods:A total of 4271 blood samples whose PLT reached the reviewing rules of thrombocytopenia were selected from inpatients having blood analysis in Xijing Hospital from January 1 st to June 30 th, 2019, including 2 200 males and 2 071 females,with a median age of (35±7.03) years old. The smears for these cases were made, stained by Wright-Giemsa, and examined to capture PLT clumps by digital cell morphology system and manual microscope separately. The digital cell analysis system (hereinafter referred to as the instrument method) as an evaluation method and the microscope method as a reference method were used to calculate the positive rate of platelet clump detection and evaluate the comparison of two methods and bias assessments. The chi-square test was used to compare counting data rates. Results:Among 4, 271 samples reaching the reviewing rule of thrombocytopenia, 128 cases with platelet clumps were detected by manual microscope(initial) with a positive detection rate of 96.24%, and a total 133 of cases with PLT clumps were detected by microscope (initial+reconfirmation) with a positive detection rate of 100 %. Meanwhile, 129 cases with platelet clumps were detected by instrument method with a positive detection rate of 96.9%. There was no significant difference in terms of positive rate of PLT clumps detection between the instrumental method and the microscope method (initial) ( χ2 =0.115, P=0.73); the positive rate of clumps detection by the instrumental method was lower than microscope method (initial+reconfirmation), and the difference was statistically significant (χ 2 =4.061, P=0.04). For instrument method, the positive rate of PLT clumps detection by simultaneous observation of RBC analysis interface+PLT aggregation interface+WBC analysis interface was higher than only observation of PLT aggregation interface, and the difference was statistically significant (χ 2 =5.090, P=0.02). The average error of the deviation of PLT counting results before and after correction of the cases with PLT plumps missed by instrument method was significantly higher than microscope method (initial), and the difference was statistically significant (χ 2 =56.26, P<0.001). Conclusion:The automated digital cell morphology system has a good consistency with manual microscope(initial) in terms of the sensitivity of platelet clumps detection and can be used as a supplementary method for detecting platelet aggregation.

Objective    To investigate the short-term quality of life in patients after single-direction video-assisted thoracic surgery (VATS) for lung cancer, and explore the related influencing factors. Methods    Patients who underwent single-direction VATS for lung cancer in the Department of Thoracic Surgery, West China Hospital, Sichuan University from July 2020 to August 2021 were continuously selected. The QLQ-C30 and QLQ-LC13 were used to evaluate the quality of life of the patients after the surgery, and the influencing factors were analyzed. Results    A total of 193 patients were collected. There were 73 males aged 59.44±11.40 years, and 120 females aged 53.73±11.15 years. The QLQ-C30 score of the patients after single-direction VATS for lung cancer was 69.09±20.21 points. Univariate analysis and Pearson correlation analysis showed that age, occupation, anesthesia time, postoperative complications, postoperative antibiotic use time, postoperative hospital stay, insomnia, economic stress, hemoptysis, chest pain, dysphagia, arm or shoulder pain were associated with the quality of life (P≤0.05). The results of multiple regression analysis showed that anesthesia time, economic pressure, insomnia and chest pain had a significant impact on the overall quality of life 30 days after the surgery (P≤0.05). Conclusion    The anesthesia time, economic pressure, insomnia and chest pain are independent influencing factors for the quality of life after the VATS lung cancer surgery.

Objective:To investigate the in vitro inhibitory activity of a novel class Ⅰ and Ⅱb selective histone deacetylase (HDAC) inhibitor, purinostat mesylate (PM) , in diffuse large B-cell lymphoma and its mechanism.Methods:The 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide method was used to detect the effect of PM on cell proliferation. The effects of PM on cell cycle and apoptosis were detected by flow cytometry. The acetylation levels of HDAC substrate, cell cycle protein, apoptosis-related protein, and oncogene protein expression were detected by Western blot.Results:PM significantly inhibited the proliferation of lymphoma SUDHL-4 and SUDHL-6 cells and increased the acetylation levels of HDAC substrates H3, H4, and α-tubulin. In cell cycle experiments, PM induced G 0/G 1 phase arrest in SUDHL-4 and SUDHL-6 cells. Western blot experiment showed that PM could significantly downregulate the expression of cyclin-dependent kinases Cdk2, Cdk4, Cdk6, cyclin D1, and cyclin E and upregulate the expression of CDK inhibitor protein p21. In the apoptosis experiment, PM could induce the apoptosis of SUDHL-4 and SUDHL-6 cells. Western blot experiment demonstrated that PM promoted endogenous apoptosis by activating caspase-3 kinase and affecting antiapoptotic protein Bcl-2. In addition, PM could downregulate the expression of oncogene marker proteins MYC, IKZF1, and IKZF3. Conclusion:PM has an efficient biological activity in vitro for diffuse large B-cell lymphoma, including double-hit lymphoma, and provides valuable experimental evidence for PM in clinical treatment.

Objective:To investigate the effects of ginsenoside Rg1 (GS-Rg1) on the secretion of exosomes (MSC-Exo) and expression of angiogenesis related microRNAs (miRNAs) in mesenchymal stem cells.Methods:Human umbilical cord blood mesenchymal stem cells (hUCBMSCs) were divided into experimental group and control group. The experimental group was treated with GS-RG1 at a final concentration of 40 mg/L, while the control group was treated with phosphate buffered saline (PBS) at the same volume. Both groups were cultured for 24 h. The morphology of MSC-Exo was observed by transmission electron microscopy; the characteristic surface markers were identified by Western blot; the concentration of MSC-Exo was detected by dicootanobutyric acid protein quantification method, and the expression of 8 miRNAs related to angiogenesis in MSC-Exo was detected by reverse transcription polymerase chain reaction (RT-PCR).Results:After 24 h of incubation, MSC-Exo with a circular membrane vesicle structure was visible. MSC-Exo was positive for the expression of the characteristic surface markers CD9, CD63 and TSG101. After 24 h of intervention, the concentration of MSC-Exo protein were (1.080±0.019)μg/μl and (0.881±0.032)μg/μl in the experimental group and control group, respectively, with statistically significant difference ( P<0.01). The expression of miR-126-3p, miR-21, miR-146a-5p and miR-125b-5p in the GS-Rg1 group were significantly higher than that in the control group, while the expression of miR-16-5p was significantly lower than that in the control group (all P<0.05). Conclusions:GS-Rg1 promotes the secretion of MSC-Exo and enhances the expression of angiogenesis-related miRNAs within Exo to promote angiogenesis.

The effect of general anesthetics (GA) on the developing brain has been a research hotspot in the last two decades. Numerous studies have shown that GA could damage the developing brain and cause long-term cognitive impairment in the preclinical setting, while it remains controversial in the clinical context regarding this topic. In light of the limitations of directly translating the preclinical findings into clinical context, drawbacks of retrospective studies, lack of standard behavioral and mental assessing tools in clinical studies, and other confounding factors, it is difficult to gain an agreement on the issue. To improve the repeatability and persuasion of data in the field of GA-induced developing brain, we tried to integrally analyze the current status and progress of the topic and make it as reference for the related studies in the future.