Objective:To investigate the mechanism of natural antigen B(nAgB)to protect Immune thrombocytopenia(ITP)mouse model by influencing autophagy.Methods:Twenty-eight female BALB/c mice aged 8-10 weeks were randomly divided into four groups.7 mice of each group were immunized intraperitoneally,the control group was treated with PBS as the control group;ITP group was treated with anti-CD41 monoclonal antibody(anti-CD41Ab)only;nAgB group was treated with nAgB intraperitoneal injection for 5d;nAgB+ITP group was treated with nAgB intraperitoneal injection for 5d,then treated with anti-CD41 Ab.The peripheral blood platelet count in each group was tested;and the spleen and liver should be isolated and weighed,the organ index was calculated;qRT-PCR was used to detect spleen microtubule-associated protein 1 light chain 3(LC3),p62,Beclin-1 mRNA expression levels.Western blot was used to detect the protein expression level of spleen LC3 Ⅱ/LC3 Ⅰ,p62,Beclin-1.Results:Compared with the control group,mice in the ITP group showed a significant decrease in blood PLT count[(102.1±17.9)× 109/L vs(485.4±185.2)×109/L,P＜0.01],a significant increase in spleen index(P＜0.01),mice in the nAgB group showed a significant increase in blood PLT count,rising to(1051±127.6)× 109/L on the 3 day after modeling.Compared with the ITP group,mice in the nAgB+ITP group showed a significant increase in PLT count on the 1 day of anti-CD41 Ab administration[(428.6±131.6)× 109/L vs(102.1±17.9)×109/L,P＜0.05],however,the spleen index was significantly decreased(P＜0.05).qRT-PCR and Western blot results showed that compared with the control group,the mRNA and protein expression levels of spleen LC3,p62 and Beclin-1 were increased in the ITP group of mice(P＜0.05,P＜0.01).Compared with the ITP group,the nAgB+ITP group could significantly decrease mRNA levels of spleen LC3,p62 and Beclin-1(P＜0.05,P＜0.01),and also significantly decrease the protein expression levels of LC3 Ⅱ/LC3 Ⅰ,p62 and Beclin-1(P＜0.05,P＜0.01).Conclusion:nAgB inhibits the transcription and expression levels of autophagy-related genes and regulates immune intolerance,thereby protecting ITP mouse models.

Objective:To investigate the mechanism of natural antigen B(nAgB)to protect Immune thrombocytopenia(ITP)mouse model by influencing autophagy.Methods:Twenty-eight female BALB/c mice aged 8-10 weeks were randomly divided into four groups.7 mice of each group were immunized intraperitoneally,the control group was treated with PBS as the control group;ITP group was treated with anti-CD41 monoclonal antibody(anti-CD41Ab)only;nAgB group was treated with nAgB intraperitoneal injection for 5d;nAgB+ITP group was treated with nAgB intraperitoneal injection for 5d,then treated with anti-CD41 Ab.The peripheral blood platelet count in each group was tested;and the spleen and liver should be isolated and weighed,the organ index was calculated;qRT-PCR was used to detect spleen microtubule-associated protein 1 light chain 3(LC3),p62,Beclin-1 mRNA expression levels.Western blot was used to detect the protein expression level of spleen LC3 Ⅱ/LC3 Ⅰ,p62,Beclin-1.Results:Compared with the control group,mice in the ITP group showed a significant decrease in blood PLT count[(102.1±17.9)× 109/L vs(485.4±185.2)×109/L,P＜0.01],a significant increase in spleen index(P＜0.01),mice in the nAgB group showed a significant increase in blood PLT count,rising to(1051±127.6)× 109/L on the 3 day after modeling.Compared with the ITP group,mice in the nAgB+ITP group showed a significant increase in PLT count on the 1 day of anti-CD41 Ab administration[(428.6±131.6)× 109/L vs(102.1±17.9)×109/L,P＜0.05],however,the spleen index was significantly decreased(P＜0.05).qRT-PCR and Western blot results showed that compared with the control group,the mRNA and protein expression levels of spleen LC3,p62 and Beclin-1 were increased in the ITP group of mice(P＜0.05,P＜0.01).Compared with the ITP group,the nAgB+ITP group could significantly decrease mRNA levels of spleen LC3,p62 and Beclin-1(P＜0.05,P＜0.01),and also significantly decrease the protein expression levels of LC3 Ⅱ/LC3 Ⅰ,p62 and Beclin-1(P＜0.05,P＜0.01).Conclusion:nAgB inhibits the transcription and expression levels of autophagy-related genes and regulates immune intolerance,thereby protecting ITP mouse models.

Objective:To analyze the MYH9 gene sequence of a patient with hereditary thrombocytopenia and diffuse large B-cell lymphoma and his family members,and to explore the relationship between MYH9 gene and tumors.Methods:Peripheral blood samples were collected from the patients and their family members for complete blood count analysis.The platelet morphology was observed under microscope.The MYH9 gene sequence was analyzed by Whole Exon Sequencing and Sanger Sequencing.Results:The mutation site c.279C＞A:p.(Asn93Lys)in exon 2 of the MYH9 gene were found in patient and his family members,both presenting as thrombocytopenia.The platelet count was significantly increased after the administration of Avatrombopag.Conclusion:A novel mutation of MYH9 was found in this study,and the case was sensitive to Avatrombopag,by exploring the relationship between the MYH9 gene and tumors,suggesting that the MYH9 gene may be associated with the development of diffuse large B-cell lymphoma.

Objective:To investigate the expression of TMSB10 mRNA in primary liver can-cer and its clinical value.Methods:The primary liver cancer data set was downloaded from the TCGA database and to analyze the expression of TMSB10 mRNA in primary liver cancer.The as-sociation between TMSB10 mRNA expression and primary liver cancer patients survival rate,im-mune cell infiltration were analyzed.A total of 150 primary liver cancer tissues and adjacent tis-sues were enrolled in our hospital,and the expression of TMSB10 mRNA in primary liver cancer tissues and adjacent tissues were analyzed via immunohistochemistry.All patients with primary liver cancer were followed up and divide into poor prognosis(n=81)and good prognosis group(n=69).The Logistic regression analysis was used to analyze the risk factors for poor prognosis in pa-tients with primary liver cancer.Results:The expression of TMSB10 mRNA in primary liver can-cer tissue and normal liver tissue were 1111.239(601.842,2154.575),396.998(324.379,524.741),respectively.When compared with normal liver tissue,the TMSB10 mRNA was significantly over-expressed in primary liver cancer tissue(P＜0.001).The median survival time of the TMSB10 mRNA lower expression group was 84.4 months,while the median survival time was 49.7 months in TMSB10 mRNA higher expression group.Compared with the TMSB10 lower expression group,the survival rate of the TMSB10 higher expression group was significantly decreased(HR=1.55,P=0.039).The expression of TMSB10 mRNA in primary liver cancer tissue was significantly positively correlated with the infiltration degree of B lymphocytes,CD8+T lymphocytes,CD4+T lympho-cytes,macrophages,neutrophils,and dendritic cells.The high expression rate of TMSB10 in pri-mary liver cancer tissues was significantly higher than that in adjacent tissues(x2=103.419,P＜0.001).The Logistic analysis showed that vascular invasion,AJCC stage with Ⅲ-Ⅳ,and higher ex-pression of TMSB were all independent risk factors for the poor prognosis in patients with primary liver cancer.In poor prognosis group,when compared with TMSB10 lower expression group,the time of recurrence and metastasis in TMSB10 higher expression group was significantly shortened(t=4.456,P＜0.001).Conclusion:TMSB10 was significantly over-expressed in primary liver cancer,and TMSB can serve as a biological marker for poor prognosis in patients with primary liver cancer.

Objective:To investigate the expression level of UBE2C in liver cancer tissues and its effect on the proliferation and invasion of hepatocellular carcinoma cells HepG2 after UBE2C silencing.Methods:The data set of liver cancer was downloaded from the TCGA database.Ac-cording to the median expression level of UBE2C mRNA in liver cancer tissues,all liver cancer pa-tients were divided into UBE2C higher(n=169)and lower expression group(n=205),respectively.The expression level of UBE2C mRNA in liver cancer tissues and its relationship with the patients prognosis was analyzed.COX regression was used to analyze the influencing factors of the liver cancer patients prognosis.The human hepatocellular carcinoma cell lines(HepG2,Huh7 and SMMC-7721)and human nromal hepatic epithelial cell line(THLE-3)were selected,and the ex-pression level of UBE2C in the four cell lines were detected by Western blot and real-time fluores-cence quantitative PCR,respectively.The HepG2 cell line was protein and mRNA expression leves divided into control group,NC group and si-UBE2C group according to UBE2C silencing.The ef-fects of UBE2C silencing on proliferation and invasion of HepG2 cell line were analyzed.Results:The expression level of UBE2C mRNA in liver cancer tissues and adjacent normal liver tissues were 4.342(3.239,5.635)and 0.905(0.587,1.230),respectively.Compared with adjacent normal liver tissues,UBE2C mRNA levels in liver cancer tissues were significantly higher(P＜0.001).The UBE2C mRNA expression levels in liver cancer tissues and paired adjacent normal liver tissues were 4.266(3.342,5.054)and 0905(0.587,1.230),respectively.Compared with paired adjacent normal liver tissues,UBE2C mRNA expression levels in liver cancer tissues were significantly higher(P＜0.001).The median survival time of UBE2C mRNA higher and lower expression groups was 48.85 months and 69.38 months.Compared with the lower expression group,the median survival time of UBE2C mRNA higher expression group was significantly shortened(P=0.045).T staging(T3/T4)and UBE2C expression(higher expression)were independent risk factors for poor prognosis in patients with liver cancer(P＜0.05).Compared with human liver epithelial cell line THLE-3,UBE2C protein and mRNA were significantly higher expressed in human hepatocellular carcinoma cell line HepG2,Huh7 and SMMC-7721(P＜0.05).The expression level of UBE2C protein and mRNA expression was the most significant in human hepatocellular carcinoma cell line HepG2 relative to cell line Huh7 and SMMC-7721.The CCK-8 results show that the cell proliferation rate in si-UBE2C group were significantly decreased protein and mRNA expression levels compared to control group and NC group at 72 h and 96 h,and the differences were significant(P＜0.05).The number of invasive cells in control group,NC group and si-UBE2C group were(23.12±3.45),(24.33±2.83)and(10.21±1.14),respectively.Compared with control group and NC group,the number of invasive cells in si-UBE2C group was significantly decreased(P＜0.05).Conclusion:UBE2C was higher expressed in liver cancer,and can be used as a biomarker for poor prognosis of patients with liver cancer.After silencing of UBE2C gene can significantly inhibit proliferation and invasion of HepG2.

Objective:To investigate the expression of TMSB10 mRNA in primary liver can-cer and its clinical value.Methods:The primary liver cancer data set was downloaded from the TCGA database and to analyze the expression of TMSB10 mRNA in primary liver cancer.The as-sociation between TMSB10 mRNA expression and primary liver cancer patients survival rate,im-mune cell infiltration were analyzed.A total of 150 primary liver cancer tissues and adjacent tis-sues were enrolled in our hospital,and the expression of TMSB10 mRNA in primary liver cancer tissues and adjacent tissues were analyzed via immunohistochemistry.All patients with primary liver cancer were followed up and divide into poor prognosis(n=81)and good prognosis group(n=69).The Logistic regression analysis was used to analyze the risk factors for poor prognosis in pa-tients with primary liver cancer.Results:The expression of TMSB10 mRNA in primary liver can-cer tissue and normal liver tissue were 1111.239(601.842,2154.575),396.998(324.379,524.741),respectively.When compared with normal liver tissue,the TMSB10 mRNA was significantly over-expressed in primary liver cancer tissue(P＜0.001).The median survival time of the TMSB10 mRNA lower expression group was 84.4 months,while the median survival time was 49.7 months in TMSB10 mRNA higher expression group.Compared with the TMSB10 lower expression group,the survival rate of the TMSB10 higher expression group was significantly decreased(HR=1.55,P=0.039).The expression of TMSB10 mRNA in primary liver cancer tissue was significantly positively correlated with the infiltration degree of B lymphocytes,CD8+T lymphocytes,CD4+T lympho-cytes,macrophages,neutrophils,and dendritic cells.The high expression rate of TMSB10 in pri-mary liver cancer tissues was significantly higher than that in adjacent tissues(x2=103.419,P＜0.001).The Logistic analysis showed that vascular invasion,AJCC stage with Ⅲ-Ⅳ,and higher ex-pression of TMSB were all independent risk factors for the poor prognosis in patients with primary liver cancer.In poor prognosis group,when compared with TMSB10 lower expression group,the time of recurrence and metastasis in TMSB10 higher expression group was significantly shortened(t=4.456,P＜0.001).Conclusion:TMSB10 was significantly over-expressed in primary liver cancer,and TMSB can serve as a biological marker for poor prognosis in patients with primary liver cancer.

Humans ; Gemcitabine ; Neoplasms

Abstract：Objective To analyze the stabilities of neuraminidase（NA）in influenza vaccine at different temperatures and provide a reference for further complete understanding of overall shelf life of vaccines. Methods Monovalent bulks of influenza H1N1，H3N2 and B vaccines were stored at 4（low temperature），25（room temperature）and 37 ℃（changed temperature）for 0. 5，2，7，24 and 48 h separately，using that at 100 ℃（extreme temperature）for 1 h as control，and determined for NA activity by enzyme⁃linked lectin method. Results The NA activities of influenza H1N1 vaccines stored at 25 and 37 ℃ decreased significantly with the increasing of time. No significant decreases were observed in H3N2 and B vaccines even after storage at two non⁃storage temperatures for 48 h. However，all the NA activities of three vaccines decreased at 100 ℃. Conclusion Both H3N2 and B vaccines showed high stability at abnormal storage temperatures not more than 37 ℃，while H1N1 vaccine was relatively sensitive to the temperature for storage.

Aim To explore the effect of γ-ray on the mRNA,protein expression levels and metabolic activity level of the key drug metabolic enzyme CYP3A1 in rat liver. Methods Wistar rats were randomly divided into control group, 24 h post-radiation group and 72 h post-radiation group. The experimental group was exposed to total body irradiation of single 6 Gy γ-ray. Blood was collected from the orbital venous plexus for blood routine examination and biochemical analysis 24 h and 72 h after irradiation, and liver tissue was prepared for quantifying expression of CYP3A1 mRNA and liver-specific microRNA (miR-122-5p) through RT-PCR. The expression level of CYP3A1 protein was analyzed by Western blot, and the metabolic activity level of CYP3A1 detected by the specific substrate midazolam combined with LC-MS method. Results Com¬pared with the control group, the weights of the rats in the radiation group significantly decreased, and the number of white blood cells was markedly reduced. Simultaneously, the activities of alanine aminotrans-ferase and alkaline phosphatase continuously descended, as well as the levels of total bilirubin and bile acid significantly increased, which indicated that the liver may be damaged after radiation. The relative expression of CYP3A1 mRNA continued to increase significantly 24 h and 72 h after irradiation. CYP3A1 protein expression and metabolic activity levels showed an obvious increasing trend 24 h after irradiation, and rose significantly 72 h after irradiation compared with the control group. At the same time, the expression of miR-122-5p in liver of rats in the 24 h and 72 h post-radiation group continued to decrease rapidly compared with the control group. Conclusions γ-ray radiation may arouse damage effect on liver, which leads to the continuous up-regulation of the mRNA and protein expression levels of the capital metabolic enzyme CYP3A1 in liver tissue, as well as the elevation of the metabolic activity level. The regulatory mechanism might be related to miR-122-5p.

Objectives: To investigated the safety and efficacy of treating patients with acute non-ST-segment elevation myocardial infarction (NSTEMI) and elevated levels of N-terminal pro-hormone B-type natriuretic peptide (NT-proBNP) with levosimendan within 24 hours of first medical contact (FMC). Methods: This multicenter, open-label, block-randomized controlled trial (NCT03189901) investigated the safety and efficacy of levosimendan as an early management strategy of acute heart failure (EMS-AHF) for patients with NSTEMI and high NT-proBNP levels. This study included 255 patients with NSTEMI and elevated NT-proBNP levels, including 142 males and 113 females with a median age of 65 (58-70) years, and were admitted in the emergency or outpatient departments at 14 medical centers in China between October 2017 and October 2021. The patients were randomly divided into a levosimendan group (n=129) and a control group (n=126). The primary outcome measure was NT-proBNP levels on day 3 of treatment and changes in the NT-proBNP levels from baseline on day 5 after randomization. The secondary outcome measures included the proportion of patients with more than 30% reduction in NT-proBNP levels from baseline, major adverse cardiovascular events (MACE) during hospitalization and at 6 months after hospitalization, safety during the treatment, and health economics indices. The measurement data parameters between groups were compared using the t-test or the non-parametric test. The count data parameters were compared between groups using the χ² test. Results: On day 3, the NT-proBNP levels in the levosimendan group were lower than the control group but were statistically insignificant [866 (455, 1 960) vs. 1 118 (459, 2 417) ng/L, Z=-1.25,P=0.21]. However, on day 5, changes in the NT-proBNP levels from baseline in the levosimendan group were significantly higher than the control group [67.6% (33.8%,82.5%)vs.54.8% (7.3%,77.9%), Z=-2.14, P=0.03]. There were no significant differences in the proportion of patients with more than 30% reduction in the NT-proBNP levels on day 5 between the levosimendan and the control groups [77.5% (100/129) vs. 69.0% (87/126), χ²=2.34, P=0.13]. Furthermore, incidences of MACE did not show any significant differences between the two groups during hospitalization [4.7% (6/129) vs. 7.1% (9/126), χ²=0.72, P=0.40] and at 6 months [14.7% (19/129) vs. 12.7% (16/126), χ²=0.22, P=0.64]. Four cardiac deaths were reported in the control group during hospitalization [0 (0/129) vs. 3.2% (4/126), P=0.06]. However, 6-month survival rates were comparable between the two groups (log-rank test, P=0.18). Moreover, adverse events or serious adverse events such as shock, ventricular fibrillation, and ventricular tachycardia were not reported in both the groups during levosimendan treatment (days 0-1). The total cost of hospitalization [34 591.00(15 527.46,59 324.80) vs. 37 144.65(16 066.90,63 919.00)yuan, Z=-0.26, P=0.80] and the total length of hospitalization [9 (8, 12) vs. 10 (7, 13) days, Z=0.72, P=0.72] were lower for patients in the levosimendan group compared to those in the control group, but did not show statistically significant differences. Conclusions: Early administration of levosimendan reduced NT-proBNP levels in NSTEMI patients with elevated NT-proBNP and did not increase the total cost and length of hospitalization, but did not significantly improve MACE during hospitalization or at 6 months.
Male ; Female ; Humans ; Aged ; Natriuretic Peptide, Brain ; Simendan/therapeutic use* ; Non-ST Elevated Myocardial Infarction ; Heart Failure/drug therapy* ; Peptide Fragments ; Arrhythmias, Cardiac ; Biomarkers ; Prognosis