Objective To establish a cell bank of extrahepatic cholangiocarcinoma (ECC)-derived organoids and investigate the key factors influencing the organoids generation. Methods The tumor samples from patients with portal cholangiocarcinoma (pCCA) and distal cholangiocarcinoma (dCCA) were used to isolate cells, and these cells were cultured using three-dimensional (3D) technique to establish ECC organoids. Histological characteristics of the organoids were evaluated and identified through hematoxylin-eosin (HE) and immunohistochemistry stainings. The success rates of organoids generation from different tumor types were compared. And clinical characteristics of patients between successful and failure culture groups were compared. Results The success rates of organoids establishment from pCCA and dCCA were all low, with 42.4% (14/33), 51.9% (14/27), respectively. The tumor was larger in successful group than that in failure group (P＜0.001); there was no statistical difference in tumor differentiation status, microvascular invasion, and perineural invasion between the two groups. Conclusions The successful rate of ECC-derived organoids establishment is low, and larger tumor has higher successful culture rate.

Objective:To establish a mass spectrometry method for the determination of characteristic peptides of Galli Gigerii Endothelium Corneum that can identify the authenticity of Galli Gigerii Endothelium Corneum as well as its preparations; To evaluate their quality.Methods:Ultra performance chromatography-electrospray tandem mass spectrometry (UPLC-MS/MS) with the mode of multiple reaction monitoring quantification (MRM) was employed to monitor the ion pairs of m/z 379.21(charge: +2)→571.36, m/z 379.21(charge: +2)→385.26, m/z 785.41(charge: +2)→941.51 and m/z 785.41(charge: +2)→245.08, in order to detect the Galli Gigerii Endothelium Corneum crude drug and its preparations. Results:Chicken specific peptide I and chicken specific peptide Ⅱ could be detected in the 18 batches of Galli Gigerii Endothelium Corneum from different regions, their corresponding extractions and 4 batches of prescription preparations, while the chicken specific peptides were not detected in the 8 batches of endothelium corneums from ducks, geese and pigeons.Conclusions:The method established in this study can effectively supplement the deficiencies in standards of Galli Gigerii Endothelium Corneum and its decoction pieces, improve the quality control standard, and provide a reference for the safety and effectiveness of Galli Gigerii Endothelium Corneum in clinical medication.

OBJECTIVE To investigate the effect of Jiawei Tianwang Buxin Dan(JWBXD)on insomnia in rats exposed to simulated high-altitude conditions.METHODS ① Thirty SD rats were randomly divided into the normal control,model,model+Jiawei Tianwang Buxin Dan(JWBXD,9.6 mg·kg-1),model+Tianwang Buxin Dan(TWBXD,9.6 mg·kg-1),and model+diazepam(DZP,3 mg·kg-1)groups.Rats,except for the normal control group,were subjected to a low-pressure,low-oxygen animal experimental chamber simulating a 5000 m altitude.Respective drugs were ig administrated once daily at 9:00 for seven days,and signal acquisition and sleep analysis were conducted by a wireless physiological sig-nal telemetry system.②Forty rats were randomly divided into five groups as described in ①.Through-out the experiment,the general condition and body mass of the rats were observed daily.Drug adminis-tration lasted for seven days,and grip strength was tested one hour after the final administration.ELISA was used to measure the levels of corticotropin-releasing hormone(CRH),adrenocorticotropic hor-mone(ACTH),corticosterone(CORT),and melatonin(MLT)in serum.Western blotting was performed to measure the expression levels of core clock proteins period circadian regulator 2(Per2),circadian locomotor output cycles(Clock),cryptochrome 2(Cry2),brain-muscle arnt-like protein 1(Bmal1),nuclear receptor subfamily 1,group D member 1(NR1D1),glycogen synthase kinase-3β(GSK-3β),as well as acetylserotonin O-methyltransferase(ASMT)in the hypothalamus and pineal gland,respectively.RESULTS ① Compared with the normal control group,the model group exhibited a decrease in total sleep time(P<0.01),an increase in wakefulness(P<0.01),a significant reduction in slow wave sleep(SWS)(P<0.05)and the mean bouts duration(P<0.05).Compared with the model group,both DZP and JWBXD(P<0.01)prolonged sleep time and suppressed wakefulness(P<0.01)in the hypoxic envi-ronment.DZP and JWBXD prolonged SWS(P<0.05,P<0.01),while TWBXD had no significant effect.JWBXD improved the mean bouts duration of SWS in the model rats(P<0.01),whereas no such improvement was observed in model+DZP and model+TWBXD groups.② Compared with the normal control group,the model group showed a significant decrease in forelimb grip strength(P<0.01),increased levels of serum ACTH(P<0.05),CRH,and CORT(P<0.01),and decreased MLT levels(P<0.05).The expression levels of Per2,Cry2,GSK-3β,and NR1D1 in the hypothalamus were downregu-lated(P<0.05,P<0.01),while Bmal1 and Clock were upregulated(P<0.05,P<0.01).ASMT expression in the pineal gland was decreased(P<0.05).Compared with the model group,JWBXD and TWBXD enhanced forelimb grip strength(P<0.01),reduced serum CORT and ACTH levels(P<0.05),decreased CRH levels(P<0.01),and restored MLT levels(P<0.01).JWBXD upregulated the expression levels of Per2,Cry2,GSK-3β and NR1D1 in the hypothalamus(P<0.05,P<0.01),but downregulated Bmal1 and Clock expression(P<0.05,P<0.01).TWBXD downregulated Bmal1 expression in the hypothalamus(P<0.01)and increased NR1D1 expression(P<0.05).DZP significantly enhanced the expression levels of Per2,Cry2 and NR1D1 in the hypothalamus(P<0.01).JWBXD,TWBXD and DZP improved ASMT expression in the pineal gland(P<0.05).CONCLUSION JWBXD can improve sleep structure and prolong the duration of SWS in rats exposed to simulated high-altitude conditions.The mechanisms may involve the regulation of core clock protein expressions in the hypothalamus,promotion of mela-tonin secretion,and inhibition of HPA axis hyperactivity.

Objective:To establish UPLC specific chromatogram method and content determination method of Dryopteridis Crassirhizomatis Rhizoma; To comprehensively evaluate the Dryopteridis Crassirhizomatis Rhizoma from different producing areas.Methods:UPLC method was used to establish the specific chromatogram of 15 batches of Dryopteridis Crassirhizomatis Rhizoma from different producing areas. The quality was evaluated by cluster analysis and partial least square analysis. The content of total flavonoids was determined by ultraviolet spectrophotometry.Results:The established specific chromatogram identified a total of 21 common peaks, and identified 4 components through comparison with control samples. The clustering and partial least squares analysis showed that the samples from Anshan City in Liaoning Province, Yanbian Korean Autonomous Prefecture in Jilin Province, Jiamusi City in Heilongjiang Province, Jilin City in Jilin Province had a certain consistency. The results of total flavone content determination showed that the content of samples from different producing areas was different.Conclusion:The established UPLC characteristic chromatogram and content determination method of total flavone content of Dryopteridis Crassirhizomatis Rhizoma can reflect the quality of Dryopteridis Crassirhizomatis Rhizoma in different places. The quality of samples from different origins is similar, but the contents of the three producing areas fluctuated greatly.

Objective:To establish a modified controlled abciximab and device investigation to lower late angioplasty complication (CADILLAC) score, and to compare the predictive value of modified CADILLAC score, the global registry of acute coronary event (GRACE) score and the thrombolysis in myocardial infarction (TIMI) score in predicting the risk of short-term death after percutaneous coronary intervention (PCI) in patients with acute ST segment elevation myocardial infarction (STEMI).Methods:A retrospective study was conducted. The clinical data of 169 STEMI patients under going PCI admitted to the department of cardiology of Guizhou Provincial People's Hospital from September 2019 to December 2020 through emergency chest pain fast track were enrolled. A multivariate Logistic regression analysis was used to screen the factors closely related to the mortality risk within 30 days of STEMI, and a modified CADILLAC scoring system was established by referring to CADILLAC scoring settings. The score of modified CADILLAC, GRACE and TIMI scores of patients were calculated after admission, and the number of deaths due to cardiovascular disease (CVD) within 30 days after onset was recorded. The receiver operating characteristic curve (ROC curve) was used to evaluate the predictive value of three scoring systems on the risk of death within 30 days after PCI in patients with STEMI.Results:In 169 STEMI patients, 16 patients died of CVD within 30 days after PCI, and the actual case mortality was 9.47%. Multivariate Logistic regression analysis showed that age > 75 years old, cardiac function Killip ≥ Grade Ⅲ, ventricular arrhythmia, ST segment elevation ≥ 0.2 mV, cardiac troponin I (cTnI) increase, systolic blood pressure (SBP) < 90 mmHg (1 mmHg ≈ 0.133 kPa) were all independent predictors of death after PCI in STEMI patients. The improved CADILLAC scoring system was constructed based on the above predictive factors combined with left ventricular ejection fraction (LVEF) less than 0.40. The GRACE, TIMI and modified CADILLAC scores of dead patients were significantly higher than those of survival patients (GRACE score: 197.60±31.83 vs. 149.81±36.72, TIMI score: 11.21±2.13 vs. 7.27±1.97, modified CADILLAC score: 12.60±2.52 vs. 6.96±2.17, all P < 0.05). The higher the risk stratification of the three scores, the higher the mortality of patients with CVD within 30 days after PCI [the mortality of patients with low, medium and high risk in GRACE score were 2.41% (2/83), 9.61% (5/52) and 26.47% (9/34); the mortality of patients with low, medium and high risk in TIMI score were 3.12% (3/96), 12.82% (5/39) and 23.53% (8/34); and the mortality of patients with low, medium and high risk in modified CADILLAC score were 3.19% (3/94), 7.69% (4/52) and 39.13% (9/23), respectively, all P < 0.01]. The area under the ROC curve (AUC) of the GRACE, TIMI and the modified CADILLAC scores predicting the risk of death 30 days after PCI in STEMI patients were 0.855 [95% confidence interval (95% CI) was 0.702-0.923], 0.725 (95% CI was 0.666-0.812) and 0.882 (95% CI was 0.732-0.936), respectively, all P = 0.000; the sensitivity of its prediction accuracy were 81.59%, 78.65% and 89.26%, and the specificity were 78.62%, 57.12% and 75.54%, respectively. Conclusions:The GRACE and the modified CADILLAC scores have predictive value for the short-term mortality risk of STEMI patients after PCI, and the modified CADILLAC score is more accurate. But the TIMI score has a poor predictive effect on the short-term mortality risk of STEMI patients after PCI.

Post-extubation dysphagia is a common problem in tracheal intubation patients. Timely identification of post-extubation dysphagia is helpful for clinical medical staff to adjust nursing strategies and improve patients' prognosis. This paper reviews the screening and evaluation tools of post-extubation dysphagia in tracheal intubation patients, so as to provide a reference for medical staff to identify post-extubation dysphagia patients in early stage.

Objective:Based on network pharmacology and molecular docking to explore the targets and mechanism of Xiaoyong Sanjie Formula treating Non-Puerperal Mastitis (NPM).Methods:By retrieving the active components and the corresponding target information of each component in Xiaoyong Sanjie Formula with Pharmacology Database and Analysis Platform of Chinese Medicine System (TCMSP), and NPM-related genes in database like GeneCard, OMIM, PharmGkb, TTD, and DrugBank, the data of the core targets of Xiaoyong Sanjie Formula and disease-related genes was compared to obtain intersecting genes, and the STRING database was used to analyze the protein interaction network and find the core genes. With the help of Cytoscape 3.8.0, the active ingredient-target-pathway regulation network diagram of Xiaoyong Sanjie Formula for the treatment of NPM was established. The R language pack was used to enrich the targets with GO function and KEGG pathway enrichment, and the potential targets and mechanism of Xiaoyong Sanjie Formula in the treatment of NPM were explored. Finally, molecular docking verification was carried out to analyze the effecacy of key components and potential core targets of Xiaoyong Sanjie Formula.Results:Network pharmacological analysis showed that there were 47 active component and 1 692 NPM-related potential targets in Xiaoyong Sanjie Formula, and 235 core targets of NPM in the treatment of Xiaoyong Sanjie Formula. The key components of Xiaoyong Sanjie Formula in the treatment of NPM include Quercetin, Naringenin, Kaempferol, Diosgenin, Luteolin, etc., with the core targets of intercellular adhesion molecule-1 (ICAM-1), vascular endothelial growth factor (VEGFA), tumor necrosis factor (TNF), interleukin-6 (IL-6), Epidermal growth factor receptor (EGFR), interleukin-1β (IL-1B), chemokine-8 (CXCL8), chemokine-2 (CCL2), etc. GO enrichment obtained 1 492 biological process entries. The KEGG pathway is enriched to obtain 105 pathways, including the TNF signaling pathway, the PI3K-Akt signaling pathway, the NF-kappa B signaling pathway, and the JAK-STAT signaling pathway, IL-17 signaling pathway, C-type lectin receptor signaling pathway, etc. The final molecular docking verified that the key active ingredients of Xiaoyong Sanjie Formula could bind with the potential core targets closely.Conclusion:Xiaoyong Sanjie Formula can treat NPM with multi-component, multi-target characteristics,which plays a role of treating NPM through signaling pathways such as immuno-inflammatory response, the metabolism of the medicine, cellular adaptive stress response, and vascular function regulation.

OBJECTIVE： To investigate in vitro antibacterial activity of the extracts from the root， stem and leaves of Psychotria rubra， and to investigate its mechanism of antibacterial effects. METHODS： 95％ ethanol extracts from the root， stem and leaves of P. rubra were used to prepare solution with mass concentration of 100 mg/mL （calculated by extract）. Using as penicillin sodium （0.5 mg/mL） and streptomycin sulfate （128 mg/mL） as positive control， plat stiletto method was used to determine antibacterial effects of the extracts from different parts of Staphylococcus aureus， Escherichia coli， Pseudomonas aeruginosa， Bacillus subtilis， Micrococcus luteus and Enterococcus faecalis. Using systematic solvent extraction method， the petroleum ether， ethyl acetate and n-butyl alcohol were used to extract antibacterial activity parts from P. rubra as ad to obtain the extracts of corresponding polar parts. After preparing 100 mg/mL drug solution （calculated by extract）， antibacterial effects of above 6 kinds of bacteria were investigated. The micro-broth dilution method and agar culture medium plate method were used to determine minimum inhibitory concentration （MIC） and minimum bactericidal concentration （MBC）， and screen polar parts with bacteriostasis and drug-sensitive strains. Under the concentration of 0.5MIC and MIC， the growth curves of sensitive bacteria were drawn （treated for 36 h， every 4 h）； the contents of alkaline phosphatase （AKP） and soluble protein were detected in suspension （treated for 10 h， every 2 h）； bouillon culture medium instead of the extract as blank control. RESULTS： 100 mg/mL ethanol extracts from the root， stem and leaves of P. rubra showed good antibacterial effect to above 6 kinds of bacteria， in descending order as stem＞leaves＞root. Among different polar parts from the stem of P. rubra， antibacterial effect of the ethyl acetate extract was best， especially for the S. aureus， the diameter of inhibition zone reached （38.93±0.12） mm， and both MIC and MBC were 0.39 mg/mL. The ethyl acetate extract could significantly inhibit the proliferation of S. aureus， and its alkaline phosphatase and protein content in suspension were increased significantly compared with blank control （P＜0.05）. CONCLUSIONS： The stem from P. rubra is effective antibacterial parts， and exhibit good antibacterial activity to 6 kinds of common pathogens. The ethyl acetate extract from the stem of P. rubra shows strongest antibacterial effect on S. aureus， and could gradually destroy the integrity of cell wall and cell membrane.

OBJECTIVE: To verify the relationship between THI and VAS scores in patients with sudden hearing loss and tinnitus in a prospective study. METHOD: Forty-four patients classified their tinnitus according to VAS and THI before and after treatment, and both scores were compared through the Pearson's correlation coefficient test and Nonparametric tests. RESULT: There was a correlation between VAS and THI scores in patients with sudden hearing loss and tinnitus. Tinnitus assessment scores were significantly reduced after treatment. Decreasing of THI and VAS scores were not depended on frequency and loudness of tinnitus and severity of deafness, but therapeutic effect of sudden sensorineural hearing loss. CONCLUSION THI and VAS scores are useful for assessment of tinnitus in patients with sudden hearing loss and tinnitus.
Audiometry ; Deafness ; Hearing Loss, Sensorineural ; complications ; Hearing Loss, Sudden ; complications ; Humans ; Prospective Studies ; Tinnitus ; complications

According to medicine high special educational reform’s mentality,we have extracted the knowledge points closely related to basic medical curriculum and the nursing spe-cialty,integrated the basic medical curriculum as "the basis of medical morphology"and"basic medical functions"and have it implemented in the nursing specialty in the three-year program.By doing so,we have gained some experience and provide practical basis for further integration of basic medical courses.