OBJECTIVE To analyze the use status and development trend of proton pump inhibitors(PPIs) in 33 hospitals in Wuhan, Hubei Province after the implementation of the national centralized drug procurement(NCDP) policy, and to provide reference for promoting the subsequent rational use of NCDP drugs and improving related policies. METHODS To make statistics and analysis of purchasing amount of PPIs, defined daily dose system(DDDs), defined daily dose consumption(DDDc) and utilization rate of 33 hospitals in Wuhan in 2019 and 2022. RESULTS After the implementation of the NCDP policy, the total purchasing amount of PPIs decreased by 53.6%, DDDs decreased by 15.4%, DDDc decreased by 45.2%, and the utilization rate of PPIs injectable dosage forms decreased by 12.6%. After NCDP, the highest growth rate of oral dosage forms was omeprazole(5.7%), followed by rabeprazole(5.0%), while injectable dosage forms showed a significant difference in utilization rate, with a significant decline in NCDP varieties and a significant increase in non-NCDP varieties. The overall NCDP utilization rate of PPIs in Wuhan was 64.9%, with little difference among hospitals of different grades. CONCLUSION The NCDP policy achieves the purpose of reducing the drug cost of patients and improving the accessibility of drugs, and is more optimized in the selection of dosage forms, which is in line with the policy expectations overall; but the quantity and price of PPIs in Wuhan decreased after NCDP, and highlighted a certain tendency in the selection of varieties. In the future, we still need to optimize measures to guide clinical priority in the selection of NCDP drugs, to ensure and improve the implementation of NCDP policy.

Objective:To explore the changes of immune microenvironment and prognosis of bladder cancer patients with positive urinary nuclear matrix protein 22 (NMP22).Methods:Retrospective analysis was made on 86 patients who were diagnosed with bladder cancer in Xuzhou Central Hospital from January 2019 to September 2020. All patients were tested for urinary NMP22 by colloidal gold method. The patients with positive test results were NMP22 positive group, and the patients with negative test results were NMP22 negative group. The expression of CD8, programmed cell death-ligand 1 (PD-L1), programmed cell death protein-1 (PD-1) and PanCK were detected by multiple fluorescent immunohistochemical method on the pathological tissue sections of all enrolled patients with bladder cancer after surgery. Follow-up data of enrolled patients were collected after discharge, and univariate and multivariate Cox analysis was performed on the follow-up data.Results:There were 69 patients in the NMP22 positive group and 17 patients in the NMP22 negative group. The percentage of CD8 and PD-L1 positive cells in NMP22 positive group was significantly higher than that in NMP22 negative group, and the difference was statistically significant (all P<0.05). Univariate analysis showed that tumor stage was correlated with bladder cancer progression ( HR=2.67, P=0.017). Multivariate analysis showed that positive NMP22 was significantly correlated with bladder cancer recurrence and disease progression (all P<0.05). Conclusions:The density of CD8 + T cells and PD-L1 in tumor parenchyma of urinary NMP22 positive bladder cancer patients was higher than that of NMP22 negative patients. Urinary NMP22 positive can be one of the bad prognostic factors of bladder cancer, and the patients with NMP22 positive should strengthen reexamination.

OBJECTIVE: To investigate the protective effect of protein kinase C (PKC) inhibitor rottlerin on rat renal vascular endothelial injury induced by lipopolysaccharide (LPS). METHODS: Rat renal microvascular endothelial cells cultured for 3-6 generations were divided into three groups according to random number table: blank control group in which cells were not challenged, LPS group in which cells were only stimulated by LPS 10 mg/L for 24 hours, and PKC inhibitor group in which cells were treated with PKC inhibitor rottlerin 2 μmol/L 30 minutes before LPS stimulation. The levels of tumor necrosis factor-α (TNF-α) and interleukins (IL-1β, IL-8) were determined by enzyme-linked immunosorbent assay (ELISA). Monolayer permeability was determined by Transwell assay. The expressions of PKC, RhoA and vascular endothelial-cadherin (VE-cadherin) were detected by Western Blot. The morphological characteristic and distribution of F-actin was measured by laser confocal fluorescence microscope. RESULTS: Compared with blank control group, the levels of inflammatory cytokines at 24 hours after 10 mg/L LPS stimulation were significantly increased in LPS group [TNF-α (ng/L): 397.3±25.4 vs. 46.8±8.9, IL-1β (ng/L): 76.7±11.2 vs. 12.6±3.2, IL-8 (ng/L): 574.5±31.4 vs. 73.2±9.6, all P < 0.05], the permeability of endothelial cells was significantly increased (A value: 1.32±0.03 vs. 0.36±0.02, P < 0.05), while the expressions of PKC and RhoA were significantly up-regulated (PKC/β-actin: 0.88±0.02 vs. 0.61±0.03, RhoA/β-actin: 0.96±0.01 vs. 0.49±0.03, both P < 0.05), VE-cadherin expression was significantly down-regulated (VE-cadherin/β-actin: 0.51±0.01 vs. 0.72±0.04, P < 0.05), and the F-actin distribution disorder had obvious stress fiber formation. Compared with LPS group, the levels of inflammatory cytokines were significantly lowered in PKC inhibitor group [TNF-α (ng/L): 127.4±14.6 vs. 397.3±25.4, IL-1β(ng/L): 43.2±7.8 vs. 76.7±11.2, IL-8 (ng/L): 212.7±18.2 vs. 574.5±31.4, all P < 0.05], the permeability of endothelial cells was significantly decreased (A value: 0.81±0.02 vs. 1.32±0.03, P < 0.05), the expressions of PKC and RhoA were significantly down-regulated (PKC/β-actin: 0.44±0.03 vs. 0.88±0.02, RhoA/β-actin: 0.63±0.05 vs. 0.96±0.01, both P < 0.05), the VE-cadherin expression was significantly up-regulated (VE-cadherin/β-actin: 0.69±0.03 vs. 0.51±0.01, P < 0.05), and the F-actin remodeling and stress fiber formation were significantly reduced. CONCLUSIONS PKC inhibitor could significantly attenuate the damage of vascular endothelial barrier induced by LPS, and plays an important role in endothelial cell barrier.
Acute Kidney Injury/prevention & control* ; Animals ; Endothelium, Vascular/drug effects* ; Interleukin-1beta ; Lipopolysaccharides/toxicity* ; Protein Kinase C/antagonists & inhibitors* ; Protein Kinase Inhibitors/pharmacology* ; Random Allocation ; Rats

Objective: To investigate the tumor-associated protein molecules carried by plasma exosomes of patients with lung squamous cell carcinoma before treatment and analyze their value as clinical markers. Methods: Exosomes from 2 patients with lung squamous cell carcinoma before treatment and 2 healthy controls were collected by ultracentrifugation. Proteomics was applied to analyze the protein expression profiles of exosomes. Candidate molecules were verified in another 30 exosomes samples from lung squamous cell carcinoma and healthy controls using enzyme-linked immunosorbent assay (ELISA). Results: Electron microscopy and particle-counting assay showed that high-quality exosomes were collected. The number of exosomes distributed from 45 to 135 nm in 2 cases of lung cancer patients were 7.89×10¹¹/ml and 9.71×10¹¹/ml, respectively, significantly higher than 2.76×10¹¹/ml and 1.41×10¹¹/ml in healthy controls. Proteomic analysis showed that proteins of exosomes in lung squamous cell carcinoma patients were very different from those of healthy controls, and some proteins are related to important functions in tumor progression. 14-3-3ζ from exosomes was selected and further verified as a marker, and the area under the receiver operating characteristic curve (ROC) was 0.68. The sensitivity and specificity of 14-3-3 ζ from exosomes were 60.0% and 80.0%, respectively, suggested that it could be used as a diagnostic marker for lung squamous cell carcinoma. Conclusion The exosome counts in plasma and the protein molecules from exosomes, such as 14-3-3ζ, are closely related to the tumorigenesis, which can be used to assist clinical diagnosis of lung squamous cell carcinoma patients.

Objective To study the differential diagnostic efficacy of α-enolase ( ENO1 ) autoantibodies in lung adenocarcinoma , benign pulmonary disease , and normal individuals , and to evaluate the improvement of the diagnostic efficiency of existing markers by establishing a binary logistic regression model.Methods This was a case-control study.Participants were from the public health welfare program led by the National Cancer Center/Chinese Academy of Medical Sciences Peking Union Medical College Cancer Hospital.Serum samples were collected June 2014 to June 2017 including 60 patients with lung adenocarcinoma , 50 patients with benign lung diseases , and 90 healthy controls.Luminex MAGPIX platform was applied to detect serum ENO1 autoantibodies, CEA and Cyfra21-1 proteins.The receiver operating characteristic curve (ROC)analysis and binary logistic regression were used to evaluate the performance and build diagnostic model.Results The median level of serum ENO1 autoantibody in patients with lung adenocarcinoma was 918.5 ( 665.5-2 043.3 ), which was significantly higher than that in the normal individuals (722.5, 585.5-921.8, Z＝-3.113, P＝0.002) and benign lung disease patients (693.0, 501.4-973.3, Z＝-3.395, P＝0.001).And no significant differences between benign disease groups and normal individuals (Z＝-1.155, P＝0.248).ROC was plotted, and the area under the curve (AUC) of ENO1 autoantibodies was 0.664 (95% confidence interval : 0.576-0.752), while the AUCs of existing diagnostic marker CEA and Cyfra21-1 were 0.680 (95% confidence interval : 0.594-0.767) and 0.617 (95% confidence interval: 0.532-0.703).A joint diagnostic model including ENO1 and CEA was built with an AUC of 0.757 (95%confidence interval : 0.675-0.838).The diagnostic efficacy of the model was significantly different from ENO1 autoantibodies (Z＝2.648, P＝0.008).When the specificity was 90%, the sensitivity of ENO1 autoantibodies was 38.3%, while the sensitivity of the combination with CEA was raised to 50%.Conclusion ENO1 autoantibodies could be a marker for the auxiliary diagnosis of lung adenocarcinoma, and can improve the efficacy of the existing diagnostic markers such as CEA .ENO1 has the potential use for the diagnosis and screening.

Objective To investigate the clinical significance of peripheral blood Th17 cells and their cytokines in the patients with tongue squamous cell carcinoma.Methods Sixty-six cases of tongue squamous cell carcinoma in our hospital from January 2013 to January 2016 served as the research subjects and contemporaneous 42 volunteers undergoing healthy physical examination were selected as the control group.Peripheral blood was collected in all subjects.The proportion of peripheral blood Th17 cells was detected by flow cytometry.The IL-17,TGF-β and IL-6 cytokine levels were detected by ELISA.Results The proportion of peripheral blood Th17 cells,IL-17,TGF-β and IL-6 in the observation group and the control group were (1.46±0.41)% vs.(0.31±0.12)%,(123.36±21.20)pg/mL vs.(20.76±8.95)pg/mL,(215.80±21.52)pg/mL vs.(26.90±10.41)pg/mL,(17.32±8.02)pg/mL vs.(5.85±1.49)pg/mL respectively,the differences were statistically significant(P<0.05).The proportion of peripheral blood Th17 cells,IL-17,TGF-β and IL-6 levels were increased with the increase of tongue squamous cell carcinoma clinical stage (P<0.01).The TGF-β level was positively correlated with the IL-17 level (r=0.626,P=0.021),the IL-6 level was positively correlated with the proportion of Th17 cells and IL-17 (r=0.626,0.597,P=0.021,0.034).Conclusion The increase of Th17 cells and IL-17 expression plays an important role in the development and progression of tongue squamous cell carcinoma,Th17 cells and IL-17 cytokine can become the important target spots of anti-tumor therapy.

Aim To explore the role of endoplasmic reticulum stress(ERS) in Astragaloside Ⅳ-induced cardioprotection in H9c2 cardiac cells, and to explore the potential mitochondrial mechanism.Methods Conventional culture was performed of rat heart tissue-derived H9c2 cells.Experiment was randomly divided into the control group, the ERS inducer 2-Deoxy-D-Glucose(2-DG) group, Astragaloside Ⅳ and 2-DG combination group, Astragaloside Ⅳ group.Confocal microscopy was used to observe the changes of TMRE fluorescence intensity so as to confirm the influence of ERS on the mitochondrial potential, and further speculate on the opening of the mitochondrial permeability transition pore(mPTP).Western blot analysis was applied to detect the expressions of ERS proteins GRP 78, GRP 94 and IRE1.Transmission electron microscopy was used to observe the ultrastructure of the cells.Results Different doses of 2-DG could mimic the mPTP opener atractyloside to induce the mPTP opening with the peak at 100 μmol·L-1;Astragaloside Ⅳ significantly reduced 2-DG-induced mPTP opening, the expression of GPR 78, GRP 94 and IRE1 and reduced injury of mitochondria and endoplasmic reticulum.Conclusions Endoplasmic reticulum stress could be induced by 2-DG.Astragaloside ⅳ-induced mitochondrial cardioprotection involves inhibition of the ERS through GRP 78, GRP 94 and IRE1 by prevention of the mPTP opening.

Objective To explore the effect of indoxyl sulfate (IS) on the differentiation, maturation and immunological function of human monocyte derived dendritic cells (mDCs), in order to provides evidence for mechanism of IS in atherosclerosis. Methods Human peripheral blood mononuclear cells isolated by double gradient centrifugation were cultured for immature mDCs by rhGM?CSF and rhIL?4 in vitro. All cases were randomly divided into PBS group, LPS group(1 μg/mL), IS.1 group(30 μmol/L), IS.2 group(300 μmol/L)and IS.3 group (600 μmol/L). The phenotypic maturation of mDCs was evaluated by flow cytometry (FCM) and functional maturation of mDCs was analyzed by measuring FITC?dextran uptake and ELISA. Results IS significantly upregulated the expression of CD80, CD83, CD86 and MHC II key membrane molecules on mDCs, while downregulating phagocytosis and increasing the secretion of IL?12p70 by mDCs (P<0.05). And the LPS and IS showed typical morphology with rough surface, long protrusions and fusiform. 300 μmol/L IS is the most appropriate stimulus concentration. Conclusion Stuctural, phenotypic and functional maturation of dendritic cells derived from human monocytes can be induced by indoxal sulphate at defined concentrations, which may be one of the mechanisms involved in the process of atherosclerosis.

This study was aimed to observe the effect ofIllicium verum water extract on excess-cold syndrome in rats. The intragastric administration of the decoction of gypsum, gentian, phellodendron andrhizoma anemarrhenae (2:1.2:1:1.5) was used to establish the excess-cold rat model. Effects ofI. verum water extract on general body signs of excess-cold syndrome rats were observed. The detection was made on the contents of LD, PA, TG, TSH, T4 in the serum and LD, PA, glycogen in liver tissues. The vitality of the LDH, SDH and ATPas was also detected. The results showed thatI. verum water extract can improve the general body signs of excess-cold syndrome rats. It reduced the content of TG; increased the content of PA, T4, TSH; reduced the content of LD, glycogen, and the vitality of LDH;increased the vitality of LDH, Ca2+-Mg2+-ATPase in liver tissues. It was concluded thatI. verum water extract can improve some general body signs of excess-cold syndrome rats, improve the substance and energy metabolism, and regulate the thyroid axis function, in order to achieve the warmingyang for dispelling cold effect.

Animals ; Cell Differentiation ; End Stage Liver Disease ; therapy ; Hepatocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology