Objective To establish a rapid screening method for 34 emerging contaminants in surface water by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UHPLC-Q-TOF-MS).Methods The pretreatment conditions of solid phase extraction(SPE)were op-timized by orthogonal experimental design and the surface water samples were concentrated and ex-tracted by Oasis? HLB and Oasis? MCX SPE columns in series.The extracts were separated by Kine-tex? EVO C18 column,with gradient elution of 0.1%formic acid aqueous solution and 0.1%formic acid methanol solution.Q-TOF-MS'fullscan'and'targeted MS/MS'modes were used to detect 34 emerging contaminants and to establish a database with 34 emerging contaminants precursor ion,prod-uct ion and retention times.Results The 34 emerging contaminants exhibited good linearity in the con-centration range respectively and the correlation coefficients(r)were higher than 0.97.The limit of de-tection was 0.2-10 ng/L and the recoveries were 81.2%-119.2%.The intra-day precision was 0.78%-18.70%.The method was applied to analyze multiple surface water samples and 6 emerging contaminants were detected,with a concentration range of 1.93-157.71 ng/L.Conclusion The method is simple and rapid for screening various emerging contaminants at the trace level in surface water.

Objective To investigate the toxicokinetic differences of 3,4-methylenedioxy-N-methylamphetamine(MDMA)and its metabolite 4,5-methylene dioxy amphetamine(MDA)in rats af-ter single and continuous administration of MDMA,providing reference data for the forensic identifica-tion of MDMA.Methods A total of 24 rats in the single administration group were randomly divided into 5,10 and 20 mg/kg experimental groups and the control group,with 6 rats in each group.The ex-perimental group was given intraperitoneal injection of MDMA,and the control group was given intraperi-toneal injection of the same volume of normal saline as the experimental group.The amount of 0.5 mL blood was collected from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.In the continuous administration group,24 rats were randomly divided into the experi-mental group(18 rats)and the control group(6 rats).The experimental group was given MDMA 7 d by continuous intraperitoneal injection in increments of 5,7,9,11,13,15,17 mg/kg per day,respectively,while the control group was given the same volume of normal saline as the experimental group by in-traperitoneal injection.On the eighth day,the experimental rats were randomly divided into 5,10 and 20 mg/kg dose groups,with 6 rats in each group.MDMA was injected intraperitoneally,and the con-trol group was injected intraperitoneally with the same volume of normal saline as the experimental group.On the eighth day,0.5 mL of blood was taken from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.Liquid chromatography-triple quadrupole tandem mass spectrometry was used to detect MDMA and MDA levels,and statistical software was employed for data analysis.Results In the single-administration group,peak concentrations of MDMA and MDA were reached at 5 min and 1 h after administration,respectively,with the largest detection time limit of 12 h.In the continuous administration group,peak concentrations were reached at 30 min and 1.5 h af-ter administration,respectively,with the largest detection time limit of 10 h.Nonlinear fitting equations for the concentration ratio of MDMA and MDA in plasma and administration time in the single-administration group and continuous administration group were as follows:T=10.362C-1.183,R2=0.974 6;T=7.397 3C-0.694,R2=0.961 5(T:injection time;C:concentration ratio of MDMA to MDA in plasma).Conclusions The toxicokinetic data of MDMA and its metabolite MDA in rats,obtained through single and continuous administration,including peak concentration,peak time,detection time limit,and the relationship between concentration ratio and administration time,provide a theoretical and data foundation for relevant forensic identification.

Objective To explore the effects of panobinostat on the proliferation,migration and invasion of renal carcinoma cells via targeting ADP ribosylated factor-like protein 4C(ARL4C).Methods According to different treatments,human renal carinoma 786-O cells were divided into blank group(phosphate buffer treatment),experimental group(50 nmol·L-1 panobinostat treatment),empty vector group(transfection with empty vector and no treatment),overexpression group(transfection with ARL4C overexpression vector and no treatment)and combined group(transfection with ARL4C overexpression vector and 50 nmol·L-1 panobinostat treatment).The proliferation,migration and invasion of cells were detected by methyl thiazolyl tetrazolium assay and scratch assay.Cholesterol transport levels in cells were detected by enzyme-labeled assay.The mRNA and protein levels of ARL4C in cells were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot.Results The indicators of cell proliferation in blank group,experimental group,empty vector group,overexpression group and combined group were(100.00±0)％,(61.08±5.82)％,(101.22±5.92)％,(121.94±6.63)％and(101.78±6.87)％;the indicators of cell migration were(44.59±2.49)％,(29.02±2.09)％,(42.75±2.42)％,(54.19±3.05)％and(41.91±2.75)％;the indicators of cell invasion were 264.50±6.52,182.70±5.83,257.80±7.91,322.40±8.27 and 266.80±8.15;the intracellular levels of cholesterol were(72.48±6.21),(36.48±3.44),(73.89±5.91),(89.21±8.89)and(73.06±6.92)mmol·L-1;the relative expression levels of ARL4C mRNA were 1.23±0.22,0.24±0.08,1.27±0.25,1.67±0.38 and 1.27±0.25;the relative expression levels of ARL4C protein were 1.06±0.03,0.17±0.04,1.03±0.05,1.37±0.18 and 1.05±0.08,respectively.Between the blank group and experimental group,the above indicators have statistically significant differences(all P＜0.05);compared with the blank group and empty vector group,the above indicators in overexpression group have statistically significant differences(all P＜0.05).Conclusion Panobinostat downregulate the intracellular levels of cholesterol by reducing the expression level of ARL4C in 786-O cells,thus inhibiting the proliferation,migration and invasion of renal carcinoma cells.

Objective To compare the early predictive value of different scoring systems for the severity,organ failure and complications of acute pancreatitis(AP)in elderly patients under the newly revised Atlanta criteria.Methods Patients with acute pancreatitis treated was collected.After admission,complete the computed tomography severity index(CTSI),the bedside index of severity in acute pancreatitis(BISAP),the pancreatis 3(PANC-3)and the harmlessness acute pancreatitis score(HAPS).The area under receiver operating characteristic(ROC)curve(AUC),sensitivity,specificity and Yordan's index of four scores for predicting SAP,local pancreatic complications and multiple organ failure were compared.Results The areas under the ROC curve predicted by the CTSI,BISAP,PANC-3 and HAPS scoring systems for SAP were 0.76,0.91,0.48 and 0.55;sensitivities of 75.87％,89.61％,61.18％and 78.38％;specificity of 80.29％,74.72％,67.48％and 69.69％;Yordan's index of 0.56,0.64,0.29 and 0.48,respectively.The AUC of CTSI,BISAP,PANC-3 and HAPS scoring systems for predicting local pancreatic complications were 0.94,0.82,0.59 and 0.64;sensitivity of 74.59％,68.23％,71.11％and 69.28％;specificity of 93.88％,83.01％,78.59％and 76.46％;Yordan's index were 0.68,0.51,0.50 and 0.46,respectively.The AUC of CTSI,BISAP,PANC-3 and HAPS scoring systems for predicting multiple organ failure were 0.60,0.84,0.64 and 0.80,sensitivities were 54.18％,74.82％,58.59％and 65.67％,specificity were 76.11％,77.20％,72.68％and 89.36％,Jordan's indices were 0.30,0.52,0.31 and 0.55,respectively.Conclusion BISAP score is higher than CTSI,HAPS and PANC-3 scoring system in predicting the accuracy of sap and the risk of multiple organ failure.

Objective To investigate the effect and mechanism of Heixiaoyao powder in regulating mitogen-activated protein kinase phosphatase-1(MKP-1)/c-Jun N-terminal kinase(JNK)signaling pathway on the level of Tau protein and neuroinflammation in the hippocampus of Alzheimer's disease(AD)rats.Methods Male Wistar rats with SPF grade were randomly divided into blank,sham-operation,model,control and experimental-L,-M,-H groups with 10 rats per group.In addition to the blank and sham-operation groups,the other 5 groups of rats were injected with β-amyloid 1-42(Aβ1-42)solution in bilateral hippocampus to replicate AD rat model,and the sham-operation group was injected with the same amount of 0.9％NaCl in the same way.Animals successfully replicated in the model were randomly divided into model group,control group(0.5 mg·kg-1 donepezil hydrochloride)and experimental-L,-M,-H groups(3.82,7.65,15.30 g·kg-1 Heixiaoyao powder decoction).The blank,sham-operation and model groups were given equal volume of 0.9％NaCl by gavage.The drug was given by gavage once a day for 42 days.Morris water maze was used to detect the learning and memory ability of rats.The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in hippocampus were detected by enzyme-linked immunosorbent assay.Western blot was used to detect the expression of MKP-1,phospho JNK(p-JNK)and phospho Tau(p-Tau)proteins.Results The escape latency on day 5 of the experimental-M,-H groups,control group,model group,sham-operation group and blank group were(8.28±7.67),(7.89±4.18),(7.86±2.68),(16.55±4.16),(6.46±3.30)and(3.60±1.53)s;the levels of TNF-α in the above groups were(406.56±28.44),(404.17±22.84),(402.28±28.36),(665.89±61.15),(226.44±34.84)and(218.50±30.16)pg·mL-1;IL-6 levels were(136.54±7.04),(121.67±5.19),(119.15±5.87),(166.27±8.91),(88.75±5.28)and(79.58±7.53)ng·L-1;the relative expression levels of MKP-1 protein were 2.31±0.34,2.59±0.38,2.58±0.37,1.23±0.25,2.64±0.19 and 2.84±0.18;the relative expression levels of p-JNK protein were 3.46±0.35,3.45±0.31,3.20±0.23,4.48±0.30,2.87±0.51 and 2.30±0.26;the relative expression levels of p-Tau protein were 3.46±0.33,3.24±0.48,3.09±0.31,4.85±1.06,2.69±0.34 and 2.40±0.55,respectively.Compared with the model group and the normal group,compared with the experimental group and the model group,the differences of above indexes were statistically significant(P＜0.05,P＜0.01).Conclusion Heixiaoyao powder can improve the learning and memory ability of AD rats,and its mechanism may be related to the regulation of MKP-1 and JNK proteins,thus inhibiting the phosphorylation level of Tau protein and alleviating neuroinflammation.

Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangtong Animal Husbandry,and whether vectors carry any bacterial pathogens that may cause diseases to humans,to provide scientific basis for prospective pathogen discovery and disease prevention and control. Methods Using a metataxonomics approach in concert with full-length 16S rRNA gene sequencing and operational phylogenetic unit(OPU)analysis,we characterized the species-level microbial community structure of two important vector species,biting midges and ticks,including 33 arthropod samples comprising 3,885 individuals,collected around Poyang Lake. Results A total of 662 OPUs were classified in biting midges,including 195 known species and 373 potentially new species,and 618 OPUs were classified in ticks,including 217 known species and 326 potentially new species.Surprisingly,OPUs with potentially pathogenicity were detected in both arthropod vectors,with 66 known species of biting midges reported to carry potential pathogens,including Asaia lannensis and Rickettsia bellii,compared to 50 in ticks,such as Acinetobacter lwoffii and Staphylococcus sciuri.We found that Proteobacteria was the most dominant group in both midges and ticks.Furthermore,the outcomes demonstrated that the microbiota of midges and ticks tend to be governed by a few highly abundant bacteria.Pantoea sp7 was predominant in biting midges,while Coxiella sp1 was enriched in ticks.Meanwhile,Coxiella spp.,which may be essential for the survival of Haemaphysalis longicornis Neumann,were detected in all tick samples.The identification of dominant species and pathogens of biting midges and ticks in this study serves to broaden our knowledge associated to microbes of arthropod vectors. Conclusion Biting midges and ticks carry large numbers of known and potentially novel bacteria,and carry a wide range of potentially pathogenic bacteria,which may pose a risk of infection to humans and animals.The microbial communities of midges and ticks tend to be dominated by a few highly abundant bacteria.

Objective:To explore the application of the semi-flipped classroom combined with knowledge point auction learning method in the theoretical teaching among stomatological students and its effectiveness.Methods:Undergraduates from four classes at Guanghua School of Stomatology, Sun Yat-Sen University were taken as research subjects. Students from two classes ( n=198) were taught using the semi-flipped classroom combined with the knowledge point auction learning method, while students from the other two classes ( n=172) were taught with the regular classroom teaching method as controls. Classroom quizzes were used to compare the two classes' mastery of important and difficult points, and a questionnaire was used to investigate students' feedback on the class outcomes. SPSS 22.0 was used for the independent samples t-test. Results:The mean test score of the experimental class was higher than that of the control class (88.51±11.02 vs. 80.40±8.53). The results of the questionnaire showed that students' interest in the course, comprehension and absorption of knowledge points, degree of satisfaction with the class, and mastery of the contents in sections in the experimental class were significantly higher than those in the control class (7.28±1.51 vs. 9.10±0.73, 6.92±1.44 vs. 8.69±1.62, 6.09±1.87 vs. 7.61±1.46, 6.40±1.04 vs. 7.70±0.86 ).Conclusions:The semi-flipped classroom combined with the knowledge point auction learning method is innovative and achieves good teaching outcomes. It holds promise for wider use in the theoretical teaching of various clinical subjects. To explore the application of the semi-flipped classroom combined with knowledge point auction learning method in the theoretical teaching among stomatological students and its effectiveness.

ObjectiveTriple-negative breast cancer (TNBC) is the breast cancer subtype with the worst prognosis, and lacks effective therapeutic targets. Colony stimulating factors (CSFs) are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells, playing an important role in the malignant progression of TNBC. This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes (CRGs), and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy. MethodsWe downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database. Through LASSO Cox regression analysis, we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score (CRRS). We further analyzed the correlation between CRRS and patient prognosis, clinical features, tumor microenvironment (TME) in both high-risk and low-risk groups, and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy. ResultsWe identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model. Kaplan-Meier survival curves, time-dependent receiver operating characteristic curves, and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival, and the predictive ability of CRRS prognostic model was further validated using the GEO dataset. Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients. Moreover, patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil, ipatasertib, and paclitaxel. ConclusionWe have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs, which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment. Moreover, the key genes within this model may represent potential molecular targets for future therapies of TNBC.

Liposome was used as carrier to carry triptolide and ginsenoside Rg3 in the treatment of pancreatic cancer tumor mice. The effects of liposome on the levels of CD4⁺ and CD8⁺ microenvironmental immune factors of pancreatic cancer tumor were investigated, and the tumor inhibitory effect and safety were evaluated. In this study, Pan02 cells were used to construct a tumor-bearing C57BL/6 mouse model. After 14 days of treatment, the changes in tumor volume and body weight of tumor-bearing mice were observed. The results showed that the high and low doses of liposome had significant therapeutic effect on tumor volume in the model group (P < 0.01), and the tumor inhibition rate of high doses of liposome was significantly increased compared with triptolide group (P < 0.05). Immunohistochemistry showed that compared with the model group, the tumor inhibition rate of liposome was significantly increased. The high-dose liposome group can up-regulate the ratio of immune factor CD4⁺/CD8⁺, inhibit the expression of tumor proliferation factor and promote the expression of tumor apoptosis factor, and has a high safety after pathological hematoxylin and eosin staining of liver, spleen, lung and kidney and serum factor detection. Animal welfare and experimental procedures are in accordance with the regulations of the Experimental Animal Ethics Committee of Henan University of Chinese Medicine (approval No.: DWLL202103173). This study provides a new idea for the exploration of immunotherapy for pancreatic cancer.

The toad, known for its various medicinal properties including parotid gland secretion (toad venom), dried skin, and gallbladder (toad bile), holds considerable medicinal applications as a valuable traditional Chinese animal medicine. Currently, in-depth attentions have been paid to the chemical composition and pharmacological properties of toad venom and skin; however, a lesser number of detailed analyses were concentrated on the toad bile. This review provides an overview of the chemical constituents in the bile of the Bufo genus, with a special focus on the cholestane and bufadienolides, and highlights the progress in their biosynthetic pathway and pharmacological activities. The analysis uncovers a distinct category of unsaturated Δ²² or Δ²³-C₂₇/C₂₈ bile acids in the toad gallbladder, potentially acting as key intermediaries in forming C-17 α-pyrone of bufadienolides. Furthermore, the high presence of 3α-OH configured bufadienolides in toad bile, in contrast to the common 3β-OH configured found in toad venom or skin, indicates a possible link between their minimal toxicity and the toad's self-defensive or physiological control. This review provides scientific basis for the development and utilization of toad bile resources, and provides useful reference for the discovery of lead compounds, analysis of the biosynthetic pathway of bufadienolides, and research on toad physiology.