ObjectiveTo explore the effect and mechanism of Taishan Panshi powder （TSPSP） on inhibiting oxidative stress injury in human chorionic trophoblast cells （HTR-8/SVneo）， and to uelucidate the underlying mechanism of TSPSP in the treatment of spontaneous abortion （SA）. MethodsGene differential analysis of SA was performed using the Gene Expression Omnibus （GEO） database and correlated with oxidative stress. Network pharmacology was employed to screen the active components of TSPSP， and a "Chinese medicine-component-target-disease" network was constructed to predict the mechanism of action of TSPSP. For in vitro validation experiments， HTR-8/SVneo cells were divided into blank group， model group， TSPSP-containing serum 2.5%， 5%， 10% groups， and nuclear factor E₂-related factor 2 （Nrf2） inhibitor group （ML385， 30 μmol·L^-1）. Except for the blank group， other groups were stimulated with 150 μmol·L^-1 H₂O₂ for 3 h to establish a cell oxidative stress injury model. After successful modeling， the blank group and model group were given 10% blank serum， each TSPSP-containing serum group was treated with the corresponding concentration of drug-containing serum， and the Nrf2 inhibitor group was additionally given 30 μmol·L^-1 ML385 on the basis of 10% TSPSP-containing serum. All groups of cells were continuously cultured under the above conditions for 24 h， and then samples were collected for subsequent detection. Cell viability in each group was detected by CCK-8 assay. Cell migration rate was detected by scratch test. The contents of malondialdehyde （MDA）， Fe²⁺， and Glutathione （GSH） were detected by enzyme-linked immunosorbent assay （ELISA）. Intracellular reactive oxygen species （ROS） level was detected by a fluorescent probe （DCF-DA）. The protein and mRNA expression levels of Kelch-like ECH-associated protein 1 （KEAP1）， Nrf2， and forkhead box protein O3 （FoxO3） in cells were detected by immunofluorescence （IF） and real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of KEAP1， Nrf2， FoxO3， Glutathione peroxidase 4 （GPX4）， and superoxide dismutase （SOD） in cells were detected by Western blot. ResultsThe GSE76862 and GSE22490 datasets were obtained from the GEO database. Differential gene analyses showed that the KEAP1， Nrf2， and FoxO3 genes were all associated with the disease. After matching with the oxidative stress pathway， nine significantly differential pathways were identified （P<0.05）， among which three contained the target genes Nrf2 and FoxO3. A total of 246 active ingredient targets of TSPSP and 2 804 SA-related targets were obtained through network pharmacology， and 154 potential action targets were obtained after taking the intersection. Topological analysis showed that targets such as KEAP1 and Nrf2 exhibited high degree values. GO and KEGG enrichment analyses indicated that the intersection targets were mainly involved in oxidative stress response， FOXO and MAPK signaling pathways， etc. In in vitro experiments， compared with the blank group， the cell viability in the model group was significantly decreased （P<0.01）. Compared with the model group， the cell viability in each TSPSP-containing serum group was significantly increased （P<0.01）. Compared with the 10% TSPSP-containing serum group， the cell viability in the ML385 group decreased to approximately 70% （P<0.01）. Compared with the blank group， the model group showed significantly increased contents of MDA， Fe²⁺， and ROS， decreased GSH expression （P<0.01）， significantly reduced cell migration rate （P<0.01）， and increased protein and mRNA expression levels of KEAP1 and FoxO3 （P<0.01）， while decreased protein and mRNA expression levels of Nrf2， GPX4， and SOD （P<0.01）. Compared with the model group， each TSPSP-containing serum group showed significantly decreased contents of MDA， Fe²⁺， and ROS， increased GSH expression （P<0.01）， significantly increased migration rate （P<0.01）， significantly decreased protein and mRNA expression levels of KEAP1 and FoxO3 （P<0.05， P<0.01）， and significantly increased protein and mRNA expression levels of Nrf2， GPX4， and SOD （P<0.05， P<0.01）. Compared with the 10% TSPSP-containing serum group， the ML385 group showed reversed trends in all indicators （P<0.05， P<0.01）. ConclusionTSPSP can inhibit H₂O₂-induced oxidative stress injury of trophoblast cells， and its mechanism of action may be related to the drug activating the KEAP1/Nrf2/FoxO3 signaling pathway.

ObjectiveTo explore the effect and mechanism of Taishan Panshi powder （TSPSP） on inhibiting oxidative stress injury in human chorionic trophoblast cells （HTR-8/SVneo）， and to uelucidate the underlying mechanism of TSPSP in the treatment of spontaneous abortion （SA）. MethodsGene differential analysis of SA was performed using the Gene Expression Omnibus （GEO） database and correlated with oxidative stress. Network pharmacology was employed to screen the active components of TSPSP， and a "Chinese medicine-component-target-disease" network was constructed to predict the mechanism of action of TSPSP. For in vitro validation experiments， HTR-8/SVneo cells were divided into blank group， model group， TSPSP-containing serum 2.5%， 5%， 10% groups， and nuclear factor E₂-related factor 2 （Nrf2） inhibitor group （ML385， 30 μmol·L^-1）. Except for the blank group， other groups were stimulated with 150 μmol·L^-1 H₂O₂ for 3 h to establish a cell oxidative stress injury model. After successful modeling， the blank group and model group were given 10% blank serum， each TSPSP-containing serum group was treated with the corresponding concentration of drug-containing serum， and the Nrf2 inhibitor group was additionally given 30 μmol·L^-1 ML385 on the basis of 10% TSPSP-containing serum. All groups of cells were continuously cultured under the above conditions for 24 h， and then samples were collected for subsequent detection. Cell viability in each group was detected by CCK-8 assay. Cell migration rate was detected by scratch test. The contents of malondialdehyde （MDA）， Fe²⁺， and Glutathione （GSH） were detected by enzyme-linked immunosorbent assay （ELISA）. Intracellular reactive oxygen species （ROS） level was detected by a fluorescent probe （DCF-DA）. The protein and mRNA expression levels of Kelch-like ECH-associated protein 1 （KEAP1）， Nrf2， and forkhead box protein O3 （FoxO3） in cells were detected by immunofluorescence （IF） and real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of KEAP1， Nrf2， FoxO3， Glutathione peroxidase 4 （GPX4）， and superoxide dismutase （SOD） in cells were detected by Western blot. ResultsThe GSE76862 and GSE22490 datasets were obtained from the GEO database. Differential gene analyses showed that the KEAP1， Nrf2， and FoxO3 genes were all associated with the disease. After matching with the oxidative stress pathway， nine significantly differential pathways were identified （P<0.05）， among which three contained the target genes Nrf2 and FoxO3. A total of 246 active ingredient targets of TSPSP and 2 804 SA-related targets were obtained through network pharmacology， and 154 potential action targets were obtained after taking the intersection. Topological analysis showed that targets such as KEAP1 and Nrf2 exhibited high degree values. GO and KEGG enrichment analyses indicated that the intersection targets were mainly involved in oxidative stress response， FOXO and MAPK signaling pathways， etc. In in vitro experiments， compared with the blank group， the cell viability in the model group was significantly decreased （P<0.01）. Compared with the model group， the cell viability in each TSPSP-containing serum group was significantly increased （P<0.01）. Compared with the 10% TSPSP-containing serum group， the cell viability in the ML385 group decreased to approximately 70% （P<0.01）. Compared with the blank group， the model group showed significantly increased contents of MDA， Fe²⁺， and ROS， decreased GSH expression （P<0.01）， significantly reduced cell migration rate （P<0.01）， and increased protein and mRNA expression levels of KEAP1 and FoxO3 （P<0.01）， while decreased protein and mRNA expression levels of Nrf2， GPX4， and SOD （P<0.01）. Compared with the model group， each TSPSP-containing serum group showed significantly decreased contents of MDA， Fe²⁺， and ROS， increased GSH expression （P<0.01）， significantly increased migration rate （P<0.01）， significantly decreased protein and mRNA expression levels of KEAP1 and FoxO3 （P<0.05， P<0.01）， and significantly increased protein and mRNA expression levels of Nrf2， GPX4， and SOD （P<0.05， P<0.01）. Compared with the 10% TSPSP-containing serum group， the ML385 group showed reversed trends in all indicators （P<0.05， P<0.01）. ConclusionTSPSP can inhibit H₂O₂-induced oxidative stress injury of trophoblast cells， and its mechanism of action may be related to the drug activating the KEAP1/Nrf2/FoxO3 signaling pathway.

Objective:To study the relationship between the type of hypertriglyceridemia(HTG) and the severity and prognosis of acute pancreatitis (AP).Methods:A total of 112 AP patients with HTG diagnosed and treated in Bozhou People's Hospital from June 2022 to June 2024 were retrospectively selected as the study objects, the AP severity and complication rate of patients with different types of HTG were compared. According to the discharge status of AP patients, they were divided into poor prognosis group (26 cases) and good prognosis group (86 cases). Univariate and multivariate Logistic regression were used to analyze the risk factors of poor prognosis in HTG patients with AP.Results:The main types of HTG in AP patients were type Ⅰ, type Ⅳ and type Ⅴ. The proportion of severe disease in AP patients with type Ⅰ HTG was higher than that of other HTG types, there were statistical differences ( P<0.05). The incidence of pancreatic pseudocyst and infectious pancreatic necrosis in AP patients with different HTG types had no statistical differences ( P>0.05). The incidence of systemic inflammatory response syndrome and organ failure in AP patients with HTG type Ⅰ was higher than that with other HTG types, there were statistical differences ( P<0.05). The proportion of type I HTG and severe AP in the poor prognosis group were higher than those in the good prognosis group : 88.46%(23/26) vs. 68.60%(59/86), 76.92%(20/26) vs. 13.95%(12/86); the triglycerides (TG), very low-density lipoprotein (VLDL), total cholesterol (TC) and fasting plasma glucose (FPG) in the poor prognosis group were higher than those in the good prognosis group: (17.29 ± 3.48) mmol/L vs. (12.29 ± 2.45) mmol/L, (3.04 ± 0.46) mmol/L vs.(2.46 ± 0.68) mmol/L,(6.03 ± 1.13) mmol/L vs.(5.25 ± 1.23) mmol/L, (12.01 ± 1.67) mmol/L vs. (11.14 ± 1.82)mmol/L; while albumin and blood calcium were lower than those in the good prognosis group: (36.28 ± 1.53) g/L vs. (37.33 ± 1.65) g/L, (1.65 ± 0.57) mmol/L vs. (2.04 ± 0.72) mmol/L; there were statistical differences ( P<0.05). The results of single factor analysis showed that combined HTG type, AP severity, triglyceride (TG), very low density lipoprotein (VLDL), total cholesterol (TC), fasting blood glucose (FPG), albumin and blood calcium levels were risk factors affecting the prognosis of AP patients ( P<0.05). Multivariate Logistic regression analysis showed that combined HTG type, AP severity, TG, VLDL, TC, FPG, blood calcium and albumin levels were independent risk factors affecting the prognosis of AP patients ( P<0.05). Conclusions:The type of HTG is closely related to the severity and prognosis of AP patients. Among them, AP patients with type Ⅰ HTG have a higher rate of severe disease. The combination of type Ⅰ HTG, the severity of AP, and abnormal lipid metabolism indexes are all important independent risk factors affecting the prognosis of AP patients.

Purpose To analyze the expression of SNAP23 in esophageal squamous cell carcinoma(ESCC)and its influence on invasion of ESCC cells.Methods A total of 41 cases of ESCC and paired normal adjacent tissue(NAT)were collected.The expression and localization of SNAP23 were detected by immunohistochemical EnVision method.The differences of SNAP23 expression levels between ESCC tissues and NAT tissues were compared to analyze the relationship between SNAP23 expression and clinicopathological characteristics.The expression level of SNAP23 in human immortalized esophageal epithelial cells SHEE and ESCC cell lines TE-1,TE-13 and KYSE150 were detected by Western blot.Lentiviral vector transfection technique was used to construct stable transfection cell lines with knock-down and overexpression of SNAP23 gene.The effect of SNAP23 on invasion of ESCC cell line TE-13 was evaluated by cell invasion experiment.Results The expression of SNAP23 in ESCC was significantly higher(53.7％,22/41)than that in the NAT group(19.5％,8/41),the difference was statistically significant(x2=10.303,P＜0.01).The expression level of SNAP23 in ESCC tissues was significantly correlated with maximum tumor diameter(x2=4.193,P＜0.05)and invasion depth(x2=7.264,P＜0.05),but was not correlated with patient gender,age,tumor site,tumor type,pathologic grade,vascular embolus,nerve invasion and lymph node metastasis(P＞0.05).Compared with human immortalized esophageal epithelial cells SHEE,SNAP23 was highly expressed in ESCC cell lines(P＜0.000 1).Compared with the sh-NT group,the invasion of ESCC cell line TE-13 was inhibited when SNAP23 expression was knocked down(P＜0.000 1);compared with the Vector group,the invasion of ESCC cell line TE-13 was enhanced after overexpression of SNAP23(P＜0.000 1).Conclusion SNAP23 is highly expressed in ESCC tis-sue and cell lines,and its expression level in ESCC tissues is positively correlated with tumor maximum diameter and invasion depth;SNAP23 promotes the invasion of ESCC cells.

Purpose To analyze the expression of SNAP23 in esophageal squamous cell carcinoma(ESCC)and its influence on invasion of ESCC cells.Methods A total of 41 cases of ESCC and paired normal adjacent tissue(NAT)were collected.The expression and localization of SNAP23 were detected by immunohistochemical EnVision method.The differences of SNAP23 expression levels between ESCC tissues and NAT tissues were compared to analyze the relationship between SNAP23 expression and clinicopathological characteristics.The expression level of SNAP23 in human immortalized esophageal epithelial cells SHEE and ESCC cell lines TE-1,TE-13 and KYSE150 were detected by Western blot.Lentiviral vector transfection technique was used to construct stable transfection cell lines with knock-down and overexpression of SNAP23 gene.The effect of SNAP23 on invasion of ESCC cell line TE-13 was evaluated by cell invasion experiment.Results The expression of SNAP23 in ESCC was significantly higher(53.7％,22/41)than that in the NAT group(19.5％,8/41),the difference was statistically significant(x2=10.303,P＜0.01).The expression level of SNAP23 in ESCC tissues was significantly correlated with maximum tumor diameter(x2=4.193,P＜0.05)and invasion depth(x2=7.264,P＜0.05),but was not correlated with patient gender,age,tumor site,tumor type,pathologic grade,vascular embolus,nerve invasion and lymph node metastasis(P＞0.05).Compared with human immortalized esophageal epithelial cells SHEE,SNAP23 was highly expressed in ESCC cell lines(P＜0.000 1).Compared with the sh-NT group,the invasion of ESCC cell line TE-13 was inhibited when SNAP23 expression was knocked down(P＜0.000 1);compared with the Vector group,the invasion of ESCC cell line TE-13 was enhanced after overexpression of SNAP23(P＜0.000 1).Conclusion SNAP23 is highly expressed in ESCC tis-sue and cell lines,and its expression level in ESCC tissues is positively correlated with tumor maximum diameter and invasion depth;SNAP23 promotes the invasion of ESCC cells.

Objective:To study the relationship between the type of hypertriglyceridemia(HTG) and the severity and prognosis of acute pancreatitis (AP).Methods:A total of 112 AP patients with HTG diagnosed and treated in Bozhou People's Hospital from June 2022 to June 2024 were retrospectively selected as the study objects, the AP severity and complication rate of patients with different types of HTG were compared. According to the discharge status of AP patients, they were divided into poor prognosis group (26 cases) and good prognosis group (86 cases). Univariate and multivariate Logistic regression were used to analyze the risk factors of poor prognosis in HTG patients with AP.Results:The main types of HTG in AP patients were type Ⅰ, type Ⅳ and type Ⅴ. The proportion of severe disease in AP patients with type Ⅰ HTG was higher than that of other HTG types, there were statistical differences ( P<0.05). The incidence of pancreatic pseudocyst and infectious pancreatic necrosis in AP patients with different HTG types had no statistical differences ( P>0.05). The incidence of systemic inflammatory response syndrome and organ failure in AP patients with HTG type Ⅰ was higher than that with other HTG types, there were statistical differences ( P<0.05). The proportion of type I HTG and severe AP in the poor prognosis group were higher than those in the good prognosis group : 88.46%(23/26) vs. 68.60%(59/86), 76.92%(20/26) vs. 13.95%(12/86); the triglycerides (TG), very low-density lipoprotein (VLDL), total cholesterol (TC) and fasting plasma glucose (FPG) in the poor prognosis group were higher than those in the good prognosis group: (17.29 ± 3.48) mmol/L vs. (12.29 ± 2.45) mmol/L, (3.04 ± 0.46) mmol/L vs.(2.46 ± 0.68) mmol/L,(6.03 ± 1.13) mmol/L vs.(5.25 ± 1.23) mmol/L, (12.01 ± 1.67) mmol/L vs. (11.14 ± 1.82)mmol/L; while albumin and blood calcium were lower than those in the good prognosis group: (36.28 ± 1.53) g/L vs. (37.33 ± 1.65) g/L, (1.65 ± 0.57) mmol/L vs. (2.04 ± 0.72) mmol/L; there were statistical differences ( P<0.05). The results of single factor analysis showed that combined HTG type, AP severity, triglyceride (TG), very low density lipoprotein (VLDL), total cholesterol (TC), fasting blood glucose (FPG), albumin and blood calcium levels were risk factors affecting the prognosis of AP patients ( P<0.05). Multivariate Logistic regression analysis showed that combined HTG type, AP severity, TG, VLDL, TC, FPG, blood calcium and albumin levels were independent risk factors affecting the prognosis of AP patients ( P<0.05). Conclusions:The type of HTG is closely related to the severity and prognosis of AP patients. Among them, AP patients with type Ⅰ HTG have a higher rate of severe disease. The combination of type Ⅰ HTG, the severity of AP, and abnormal lipid metabolism indexes are all important independent risk factors affecting the prognosis of AP patients.

Objective To investigate the clinical features of patients with heart failure and the safety and efficacy of noninvasive ventilator in patients with heart failure.Methods Sequentially enrolled 65 patients who were diagnosed with decompensated heart failure in Tianjin Chest Hospital Heart Center from October 2016 to October 2017 and who had acute heart failure during hospitalization requiring noninvasive ventilator,were divided into the HF-PEF group (n=19) and HF-REF group (n=46).The clinical data of the two groups and the observation indexes before and after the application of the non-invasive ventilator were compared.Results Comparing the admission data of the two groups,the proportion of patients with hypertension (57.9％ vs 21.7％,P=0.005) and LVEF(％) (53.00±4.85 vs 33.07±7.24,P＜0.01)were significantly higher in the HF-PEF group than those in the HF-REF group;LVEDD (mm) in the HFPEF group was significantly lower than that in the HF-REF group (50.00±5.23 vs 63.82±8.95,P＜0.01).In the two groups of patients with acute left heart failure,blood lactate levels (mmol/L) in the HF-PEF group (4.20±1.06 vs 2.02±0.88,P＜0.05) and systolic blood pressure (mmHg) (151.32±43.40 vs 117.90± 19.55,P＜0.05) were significantly higher than those in the HF-REF group.After the application of non-invasive ventilator,systolic blood pressure (mmHg) (34.38±9.36 vs 16.94±5.19,P=0.038) and PaCO2 (mmHg)(2.49±0.98 vs-0.06±0.00,P=0.025),and lactic acid (mmol/L) (2.06±0.67 vs 0.04±0.01,P=0.001) were significantly lower in the HF-PEF group than those in the HF-REF group.While the NT-proBNP level (ng/L) (13 064.90±1 963.83 vs 11 687.13±1 028.03,P=0.848) did not decrease significantly,and the time of non-invasive ventilator application (h)was significantly longer than that in the HF-REF group (152.74±10.61 vs 71.03±10.41,P=0.013).Conclusions Hypertension is the main cause of HF-PEF group.The systolic blood pressure and blood lactate level in HF-PEF patients with acute left heart failure are significantly higher than HF-REF patients.Non-invasive ventilator is also safe and effective for the treatment of acute left heart failure in HF-PEF patients,but HF-PEF patients with acute left heart failure have a longer clinical remission time.

Two proteins of similar molecular weight (named as ASPR-C-1 and ASPR-C-2) from the crude drug of Angelica sinensis were purified and characterized by 80% ammonium sulfate precipitation, Sephadex G-50 gel filtration chromatography, and DEAE-Sepharose anion exchange chromatography. The molecular weight of ASPR-C-1 and ASPR-C-2 on SDS-PAGE was 17.33 kDa and 17.18 kDa, respectively. They were mainly monomeric in solution, but partially formed dimers and they were glycoproteins with glycosyl content of 2.6% and 8.2%, respectively. Both ASPR-C-1 and ASPR-C-2 were identified to be members of pathogenesis-related 10 family of proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and have ribonuclease activities with the specific activity of 73.60 U/mg and 146.76 U/mg, respectively. The optimum pH of the two isoforms was similar, at about 5.6, while their optimum temperatures were different. The optimum temperature of ASPR-C-1 was 50 ℃, and that of ASPR-C-2 was 60 ℃. Both isoforms presented highest thermal stability at 60 ℃. However, ASPR-C-2 was more thermotolerant than ASPR-C-1. The latter was rapidly inactivated and retained only about 20% residual activity while the former still maintained about 80% of its original activity at a higher treatment temperature (80 to 100 ℃). In addition, Fe²⁺ had an activating effect on the ribonuclease activities of two isoforms while Ca²⁺, Mg²⁺, Zn²⁺, Mn²⁺, Ag⁺, Cu²⁺, EDTA (Elhylene diamine tetraacetic acid), dithiothreitol and sodium dodecylsulphate showed different degrees of inhibition of the enzyme activities. Our findings provide a foundation for further research on the biological function of PR-10 protein from Angelica sinensis.
Angelica sinensis ; Chromatography, Gel ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; Hydrogen-Ion Concentration ; Kinetics ; Molecular Weight ; Protein Isoforms ; Temperature

OBJECTIVE: To assess the association of 5A/6A polymorphism in the promoter region of MMP3 gene with the stability of extracellular matrix of atherosclerotic plaque. METHODS: Clinical data of 776 consecutive patients undergoing percutaneous coronary intervention (PCI) was reviewed. MMP3 gene polymorphisms and serum level of MMP3 for the second admission were collected. The target gene fragment containing MMP3 promoter region was transfected into HepG2 vector cells. The influence of the polymorphism on the expression of the MMP3 gene was determined in vitro. RESULTS: Compared with the first admission data, the proportion of mutant MMP3 genotypes (5A/5A+5A/6A) was significantly higher in patients with acute myocardial infarction (AMI) compared with the control group (37.6% vs. 24.9%, P<0.01). 64.1% of the patients carrying the 5A allele had AMI, whereas only 50.11% of those carrying the 6A allele had AMI (P<0.01). The proportion of wild-type MMP3 genotype (6A/6A) was significantly higher in the stenotic group compared with the non-restenosis group (79.5% vs. 66.5%, P<0.01). Restenosis has occurred in 9.5% of patients harboring the 5A allele compared with 16.2% in those carrying the 6A allele (P<0.01). In addition, serum level of MMP3 in the restenosis group was significantly lower than that of the non-restenosis group (P<0.01). In vitro studies confirmed that the expression of pGL2-Basic/6A was significantly lower than that of pGL2-Basic/5A. CONCLUSION The 5A/6A polymorphism in the promoter region of the MMP3 gene may influence its transcriptional activity and impact on the degradation or push-up of extracellular matrix, resulting in a difference in the stability of atherosclerosis plaques, which in turn may induce different pathological processes in AMI or restenosis after stenting.
Case-Control Studies ; Extracellular Matrix ; Genetic Predisposition to Disease ; Genotype ; Humans ; Matrix Metalloproteinase 3 ; genetics ; Percutaneous Coronary Intervention ; Plaque, Atherosclerotic ; genetics ; Polymorphism, Genetic ; Promoter Regions, Genetic