Objective To investigate the effect of different concentrations of TPh on hepatoma cells , cell viability and and its possible mechanisms of anti-tumor.Methods The inhibitory rate of hepatoma cells and cell viability on different concentrations of TPh and time were measured by MTT assay;The morphological changes of apoptosis were observed by Hoechst 33258 straining; Cell cycle distribution was evaluated by flow cytometry (FCM).The protein expression of Bcl-2 and Bax were detected by Western blot analysis.Results MTT assay showed that TPh inhibited the proliferation of HepG-2 cells in a dose-dependent manner, and the inhibitory rate increased with the increase of concentration.The inhibitory rate was 50.9% (P<0.01).The results of Hoechst 33258 staining showed that the fluorescence intensity of hepatocellular carcinoma cells was light blue in fluorescence microscopy and bright blue fluorescence in apoptotic hepatocarcinoma cells, and the apoptosis of hepatocarcinoma cells increased with the increase of drug concentration.The percentage of cells in G0/G1 phase increased with the increase of cell cycle, and the ratio of cells in S phase was decreased in G2/M phase compared with blank control group (P<0.05);Western blot results showed that compared with the blank control group, TPh inhibited the proliferation of Bcl-2 cells in a concentration-dependent manner ( P <0.05 ) , and the number of apoptotic cells increased with the increase of drug concentration (P<0.05), and the ratio of Bcl-2/Bax in the TPh group decreased significantly (P<0.05), and the expression of Bax gene increased. Conclusion TPh inhibits cell proliferation, promotes apoptosis and induces HepG-2 to block G0/G1 phase.Its mechanism may increase the expression of Bax and decrease Bcl-2 protein expression.

AIM:To predict the active compo-nents and targets of Piperlongum L.and the associ-ated signaling pathways involved in pulmonary fi-brosis using network pharmacology and molecular docking technique and evaluate the mechanism of Piperlongum L against pulmonary fibrosis by in vi-tro experiments.METHODS:The active ingredients and targets were retrieved from TCMSP,Swiss Tar-get Prediction and PubChem databases.The dis-ease-related targets were retrieved from Gene-Cards and OMIM databases.The intersection tar-gets of the drugs and disease-related targets were identified using jvenn online tool.String database was used to construct the"drug-component-tar-get"and PPI network and the networks were visual-ized using Cytoscape 3.9.1 software.GO and KEGG enrichment analysis were performed on the inter-section targets using the DAVID tool.The top 20 KEGG pathways,core targets and drug components were used to construct a"component-target-path-way"network and the network visualization was performed using Cytoscape 3.9.1 software.The in-teractions between drug compounds and the tar-gets were evaluated by molecular docking,and the docking results were visualized using Discovery stu-dio.HFL-1 cells were cultured and the effect of the drug compounds on cell viability was determined by MTT assay.The inhibition rate was then calculat-ed to determine the optimal drug concentration.HFL-1 cells were cultured in vitro and were as-signed into 4 groups:control group,TGF-β1 group,TGF-β1+LD group(LD group),TGF-β1+HD group(HD group).CCK-8 kit was used to evaluate the anti-proliferative activity of the drug compounds against HFL-1 cells at 24,48 and 72 h.Plate clone formation assay was performed to evaluate the ef-fect of drugs on the colony formation ability of HFL-1 cells.RT-qPCR and western blot were conducted to determine the effect of the compounds on the mRNA and protein expression levels of α-smooth muscle actin(α-SMA),collagen type Ⅰ(COI-Ⅰ),and collagen type Ⅲ(COI-Ⅲ)in each group.RESULTS:A total of 197 intersection targets of Piperlongum L and anti-pulmonary fibrosis were identified.The core PPI network comprised 29 nodes(targets)and 199 edges(interactions).GO functional analysis showed that the significantly enriched biological processes associated with the compounds in Piper-longum L included negative regulation of apopto-sis,signal transduction,and protein phosphoryla-tion.Significantly enriched cellular components in-cluded cytoplasm,nuclear cytoplasm,plasma mem-brane.Enriched molecular functions associated with the compounds included the same protein binding,serine/threonine/tyrosine kinase activity,and protein binding.A total of 155 significantly en-riched KEGG signaling pathways were identified,with PI3K-Akt signaling pathway was highly associ-ated with PF and was the fourth most enriched pathway.PIK3CA,MAPK3,MAPK1,MTOR,SRC,CCND1,EGFR,PRKCA,BCL2,and GSK3B had the highest connectivity in the components-target-pathway network.Piperlongine,N-(2,5-dimethoxy-phenyl)-4-methoxybenzamide,tetrahydrotanshi-none,pisigenin and piperine were the key com-pounds in Piperlongum L.The molecular docking results showed that all the compounds except N-(2,5-dimethoxyphenyl)-4-methoxybenzamide had good binding activities with interactions observed with 10 proteins.The proliferation ability of the cells in the LD group was significantly lower than that of the TGF-β1 group at 48 h and 72 h(P＜0.05).The proliferation ability of cells HD group was sig-nificantly lower than the LD group at 24,48 and 72 h.The number of clones in each drug group was significantly reduced after treatment with the drugs(P＜0.05).The mRNA and protein expression levels of α-SMA,COI-Ⅰ,COI-Ⅲ in LD and HD groups were significantly lower than the expression levels in the TGF-β1 group.The protein expression levels of p-PI3K/PI3K and p-Akt/AKT were significantly lower in the two dose groups compared with the TGF-β1 group(P＜0.01).CONCLUSION:The results showed that the effect of Piperlongum L against PF is probably through modulation of the PI3K-Akt sig-naling pathway.

Hydrofluoric acid inhalation injury is difficult to treat, despite it has low incidence. It could cause mild symptoms such as cough and sore throat, or severe symptom that may develop into life-threatening acute respiratory distress syndrome, and even rare pulmonary diseases such as reactive airway dysfunction syndrome and pulmonary alveolar proteinosis. Currently, there is no specific standard for the diagnosis and treatment of hydrofluoric acid inhalation injury. Authors summarize the incidence, injury mechanism, clinical diagnosis and treatment of hydrofluoric acid inhalation injury by searching literature at home and abroad and propose that pulse contour cardiac output monitor and extracorporeal membrane oxygenation have great application prospects in treatment of severe cases, so as to provide references for peers.

Objective:To analyze the clinical characteristics of pesticide poisoning patients and explore the risk factors of acute kidney injury (AKI) .Methods:In September 2020, the clinical data of 155 patients with pesticide poisoning in the department of nephropathy, the Affiliated Hospital of Southwest Medical University from September 2018 to August 2020 were retrospectively analyzed. The patients were divided into AKI group (44 cases) and non AKI group (111 cases) according to the occurrence of AKI. The clinical characteristics, organ or system involvement and auxiliary examination results of the two groups were analyzed. Logistic regression was used to analyze the risk factors of AKI in patients with pesticide poisoning.Results:The types of pesticides causing poisoning mainly included herbicides, insecticides and biochemical pesticides. Compared with non AKI group, patients in AKI group had higher proportion of blood purification treatment and ICU monitoring treatment ( P<0.05) , and were more likely to be complicated with acute respiratory failure, pulmonary fibrosis, myocardial injury, multiple organ dysfunction syndrome (MODS) , acute pancreatitis and coagulation abnormalities ( P<0.05) . The mortality of AKI group (18.2%, 8/14) was significantly higher than that of non AKI group (0.9%, 1/111) ( P<0.05) . Univariate analysis showed that the time from poisoning to treatment > 6 h, high WBC count, neutrophil count, alanine aminotransferase, aspartate aminotransferase, high sensitive troponin T, myoglobin and creatine kinase isoenzyme were the risk factors of AKI in patients with pesticide poisoning ( P<0.05) . Multivariate logistic regression analysis showed that the time from poisoning to treatment >6 h was an independent risk factor for AKI in patients with pesticide poisoning ( P<0.05) . Conclusion:The mortality of AKI secondary to pesticide poisoning is high. Attention should be paid to the time from poisoning to treatment, inflammatory state and changes of liver and myocardial function.

Objective:To analyze the clinical characteristics of pesticide poisoning patients and explore the risk factors of acute kidney injury (AKI) .Methods:In September 2020, the clinical data of 155 patients with pesticide poisoning in the department of nephropathy, the Affiliated Hospital of Southwest Medical University from September 2018 to August 2020 were retrospectively analyzed. The patients were divided into AKI group (44 cases) and non AKI group (111 cases) according to the occurrence of AKI. The clinical characteristics, organ or system involvement and auxiliary examination results of the two groups were analyzed. Logistic regression was used to analyze the risk factors of AKI in patients with pesticide poisoning.Results:The types of pesticides causing poisoning mainly included herbicides, insecticides and biochemical pesticides. Compared with non AKI group, patients in AKI group had higher proportion of blood purification treatment and ICU monitoring treatment ( P<0.05) , and were more likely to be complicated with acute respiratory failure, pulmonary fibrosis, myocardial injury, multiple organ dysfunction syndrome (MODS) , acute pancreatitis and coagulation abnormalities ( P<0.05) . The mortality of AKI group (18.2%, 8/14) was significantly higher than that of non AKI group (0.9%, 1/111) ( P<0.05) . Univariate analysis showed that the time from poisoning to treatment > 6 h, high WBC count, neutrophil count, alanine aminotransferase, aspartate aminotransferase, high sensitive troponin T, myoglobin and creatine kinase isoenzyme were the risk factors of AKI in patients with pesticide poisoning ( P<0.05) . Multivariate logistic regression analysis showed that the time from poisoning to treatment >6 h was an independent risk factor for AKI in patients with pesticide poisoning ( P<0.05) . Conclusion:The mortality of AKI secondary to pesticide poisoning is high. Attention should be paid to the time from poisoning to treatment, inflammatory state and changes of liver and myocardial function.

To prepare a lipid nanoparticle (LNP)-based subunit vaccine of Mycobacterium tuberculosis (Mtb) antigen EsxV and study its immunological characteristics, the LNP containing EsxV and c-di-AMP (EsxV: C: L) was prepared by thin film dispersion method, and its encapsulation rate, LNP morphology, particle size, surface charge and polyphase dispersion index were measured. BALB/c mice were immunized with EsxV: C: L by nasal drops. The levels of serum and mucosal antibodies, transcription and secretion of cytokines in lung and spleen, and the proportion of T cell subsets were detected after immunization. EsxV: C: L LNPs were obtained with uniform size and they were spherical and negatively charged. Compared with EsxV: C immunization, EsxV: C: L mucosal inoculation induced increased sIgA level in respiratory tract mucosa. Levels of IL-2 secreted from spleen and ratios of memory T cells and tissue-resident T cells in mice were also elevated. In conclusion, EsxV: C: L could induce stronger mucosal immunity and memory T cell immune responses, which may provide better protection against Mtb infection.
Animals ; Mice ; Mycobacterium tuberculosis ; Antigens, Bacterial ; Immunization ; Nanoparticles ; Vaccines, Subunit ; Mice, Inbred BALB C