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

Objective To discuss the effect of Anemarrhenae Rhizoma with different proportions on the dissolution of 6 inorganic elements(K,Ti,Sr,Ba,Ca,Mg)in Gypsum Fibrosum under different compatibility based on grey correlation analysis and technique for order preference by similarity to an ideal solution(TOPSIS)method and to provide reference for clinical application.Methods Inductively coupled plasma mass spectrometry(ICP-MS)was established for simultaneous analysis of inorganic elements K,Ti,Sr,Ba,Ca,Mg in Gypsum Fibrosum-Anemarrhenae Rhizoma.The gray correlation ananlysis and TOPSIS method were used to comprehensively analyze the changes of six inorganic elements in Gypsum Fibrosum under different compatibility.Results The findings of the ICP-MS determination of inorganic elements in Gypsum Fibrosum-Anemarrhenae Rhizoma were good in terms of linearity,precision,repeatability and stability.The average recovery was 81.78％-104.13％,and the RSD was 2.10％-4.62％(n=6).The dissolution of six inorganic elements in Gypsum Fibrosum after compatibility with different proportions of Anemarrhena Rhizoma was significantly higher than that of Gypsum Fibrosum single decoction(P＜0.05).The dissolution scores of six inorganic elements in Gypsum Fibrosum were the highest when the compatibility proportion of Gypsum Fibrosum-Anemarrhenae Rhizoma was 30∶9.Conclusion ICP-MS is a sensitive and accurate method for determining K,Ti,Sr,Ba,Ca and Mg in Gypsum Fibrosum-Anemarrhenae Rhizoma.The best compatibility ratio of Gypsum Fibrosum-Anemarrhenae Rhizoma is 30:9,which can effectively improve the dissolution of six inorganic elements in Gypsum Fibrosum,and provide the basis for the further compatibility study of Gypsum Fibrosum and Anemarrhenae Rhizoma.

Arecae Semen, as the first place among "Four South Medicines" in China, has great dual-use value of medicine and food. The research of Arecae Semen was mainly focused on the active ingredients and efficacy value, and its potential safety hazards were also concerned. Until now, there is still a lack of clear boundaries between medicine and food, resulting in its safety cannot be guaranteed. Therefore, it is of great significance to establish clear boundaries of medicine and food use and health risk assessment. In this paper, the differences of pretreatment and application methods of Arecae Semen were analyzed, and the research progress of Arecae Semen in chemical composition identification and toxicology research and safety evaluation were reviewed emphatically. Finally, the differences of quality control and safety evaluation of Arecae Semen in pharmacopoeias or standards were analyzed at home and abroad. It was expected to provide reference value for quality control, safety evaluation and international standardization research of Arecae Semen.
Areca ; China ; Drugs, Chinese Herbal/adverse effects* ; Seeds ; Semen

【Objective】 To explore the effect of prostate volume on outcomes of nerve sparing laparoscopic radical prostatectomy(nsLRP) and investigate its predictive value for postoperative continence recovery. 【Methods】 A retrospective study was conducted on 178 patients with low risk prostate cancer(PCa) who underwent nsLRP from January 2004 to June 2018. All the patients included had normal preoperative urinary function, a mean age of 66 years(range 46~88), a median PSA level of 6.9 μg/L(range 0.6~10.0) and a mean body mass index of(24±5) kg/m². The correlation of prostate volume with perioperative data was assessed by using Spearman correlation coefficient. Univariate and multivariate Cox regression analyses were employed to identify prognostic predictors for postoperative continence recovery. 【Results】 Increased prostate volume was significantly correlated with older age(rs=0.370, P<0.001), higher PSA level(rs=0.168, P=0.025), lower biopsy Gleason score(rs=-0.165, P=0.027), lower pathological T stage(rs=-0.152, P=0.042) and Gleason score(rs=-0.182, P=0.015), longer operative time(rs=0.239, P=0.001) and greater estimated blood loss(rs= 0.181, P=0.016). The continence rates in the 3rd, 6th, and 12th month after surgery were 64.0%(114/178), 88.2% (157/178), and 96.1%(171/178), respectively. Univariate and multivariate Cox analysis showed that patient age(RR= 0.528, 95% CI: 0.381~0.730) and prostate volume(RR=0.598, 95% CI: 0.412~0.869) were independent predictors of continence recovery. 【Conclusions】 Larger prostate volume was associated with older age, higher PSA level, lower tumor stage and grade, greater intraoperative blood loss and longer operative time in low risk PCa patients. Older age and increased prostate volume may independently predict poor continence recovery after nsLRP.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

OBJECTIVE To study the methodology of achieving stable co-expression of drug-metab?olizing enzymes in the HepG2 cells by the piggyBac (PB) transposon system. METHODS N-terminal attachment of enhanced green fluorscent protein plasmid (pEGFP- N2) and 2A peptide linked recombinant PB transposon plasmid containing dual-genes encoding drug metabolizing enzymes cyto?chrome P450 3A4 (CYP3A4) and CYP2C19 (pPB-CYP3A4-2A-2C19) were transfected into HepG2 cells respectively by Lipofectamine?LTX reagent, GenJetTM (Ver.Ⅱ) reagent and Neon?Transfection System reagent, which were widely used for large-sized DNA fragments transfection. 48 h later, the transfection efficiency and cell toxicity were detected and compared between the three methods so as to find a method with relatively high efficiency and low toxicity for later transfection.Then,three groups of recombinant PB transposons-single-gene transposon (PB-CYP3A4), 2A peptide linked dual-gene transposon (PB-CYP3A4-2A-2C19) and multiple single-gene transposon mixture〔PB-CYP3A4, PB-CYP2C8, PB-CYP2A6, organic anion transporting polypeptide 1B1 PB transposon (PB-OATP1B1)〕-were transfected into HepG2 cells respectively with the above established method.The puromycin (Puro)-resistant and GFP positive cell clones were picked up and further cultured. The mRNA, protein and metabolic levels of drug-metabolizing enzymes in monoclonal cell lines were detected by quantitative real-time PCR,Western blotting and high performance liquid chromatography-tandem mass spectrometry respectively after screening by Puro and green fluorescence. Comparisons of different groups were made using statistical analysis. RESULTS The comparison of three different transfection methods indi?cated that the transfection efficiency of GenJetTMwas up to(94.2±2.5)% and (89.3±3.3)%,significantly higher than those of the other two methods (P<0.01), along with lower cytotoxicity. Then GenJetTMwas chosen for later transfection. In the Puro-resistant monoclonal cell lines of single transposon PB-CYP3A4,PB-CYP3A4-2A-2C19 groups,the mRNA,protein and enzyme activity levels of drug-metabo?lizing enzymes were significantly increased respectively.The recombinant transposon (PB-CYP3A4-2A-2C19) containing 2A peptide could achieve stable and efficient co-expression of two metabolizing enzymes CYP3A4 and CYP2C19,while the expression of drug-metabolizing enzymes remained unbal?anced and random in those of multiple single-gene transposon mixture group (PB-CYP3A4, PB-CYP2C8,PB-CYP2A6,PB-OATP 1B1)(CYP3A4 was expressed in some cell clones only).CONCLUSION GenJetTM could be an effective method for the PB recombinant transposon transfection into HepG2 cells, by which the PB transposon could mediate stable expression of drug-metabolizing enzymes. In terms of multi-gene expression,a low and unbalanced expression is found by multiple transposons co-transfection method,which is different from that by virus mediated method.In contrast,mono-PB trans?poson linked by 2A peptide can achieve stable expression of multi-genes.

Objective To provide effective reference for quality analysis of the chemical composition and extraction of astragalus separation process by comparing the extract of astragalus and it’s IR spectra. Methods The saponins and flavonoids in astragalus were firstly extracted by the method of circumfluence with ethanol as solvent and the residue of ethanol-extraction was then used to extract polysaccharides by distilled water. Fourier transform infrared spectroscopy (IR) combined with second derivative infrared spectroscopy was applied to quickly identify astragalus herbs powder, water extraction of astragalus, astragalus alcohol extraction and water extraction of the residue of ethanol-extraction. Results The powder and 70% ethanol extract, 80% ethanol extract were around at 1 735 cm-1 (carbonyl stretching vibration absorption peak) has a weak, broad absorption, while the absorption peak was less obvious in boiling water extraction. So the maln components of astragalus water extraction are polysaccharides and also contaln a small amount of water-insoluble flavonoids. Alcohol extraction malnly contalns saponins and flavonoids, and flavonoid extract volume increases with increasing alcohol concentration in a certaln range.Conclusion This method can be a quick and easy identification for astragalus and it’s extraction for its chemical composition class, and provide the basis for further research quality.

Objective To analyze the clinical and pathological characteristics of 35 hepatocellular carcinoma (HCC) patients with bile duct tumor thrombi (BDTT),and to investigate the expressions of CD133,CD90,EpCAM,CK19,VEGF,and C-kit in the tumor tissues.Methods 35 HCC patients with BDTT out of 943 HCC patients who underwent surgical treatment were studied.The expressions of biomarkers in tissue specimens were determined by immunohistochemistry.35 HCC patients without BDTT were selected using the method of stratified sampling as a control group.Results In 19 of 35 patients,the diameters of the primary tumor were less than 5 cm (range 0 ～ 17 cm,average 6.9 ± 0.7 cm).When compared to the control group,most of the primary tumors were moderately to lowly differentiated (33/35,94％ vs 18/ 35,51％),had incomplete capsules (18/35,51％ vs 3/35,8％) and micro vascular invasion (29/35,83％ vs 7/35,20％).The positive expression rates of CD90,EpCAM,CK19,VEGF,CD133,and C-kit in the group of patients with HCC with BDTT and in the control group were 82.9％,77.1％,71.4％,85.7％,80.0％,80.0％ and 57.1％,54.3％,34.3％,65.7％,54.3％,51.4％,respectively.The 1-,2-,3-year postoperative survival rates of the HCC patients with BDTT were 69％,37％,20％ respectively which were worse than the HCC patients without BDTT (1-,2-,3-year postoperative survival rates were 88％,72％,62％ respectively,P ＜ 0.05).Conclusions The prognosis of HCC patients with BDTT was worse than HCC patients without BDTT.The expressions of liver stem cell biomarkers in the tumor specimens of the group of HCC patients with BDTT were higher than the control group.These findings prompt that this kind of HCC originate from liver stem ceils.

This study was purposed to explore the changes of possible angiogenetic factors other than VEGF after inhibition of NHE1 and their related mechanisms. The K562 cells were treated by NHE1 specific inhibitor cariporide, the angiogenesis factors after inhibition of NHE1 were screened by using protein chip, the IL-8 expression level after cariporide treatment was detected by real-time quantitative PCR; the K562 cells with stable interference of NHE1 were constructed, the IL-8 expression level after interference of NHE1 was detected by real-time quantitative PCR; the p38 phosphorylation level in K562 cells treated with cariporide was detected by Western blot. After treatment of K562 cells with p38 inhibitor SB203580, the IL-8 expression level was decreased by real-time quantitative PCR. The results of protein chip showed that IL-8 expression decreased after cariporide treatment. Real-time quantitative PCR confirmed this inhibitory effect. The p38 phosphorylation level increased after cariporide treatment. The down-regulation of IL-8 expression induced by cariporide treatment was partially restored after K562 cells were treated with p38 inhibitor SB203580. It is concluded that the inhibition of NHE1 can inhibit IL-8 expression through up-regulation of p38 phosphorylation.
Cation Transport Proteins ; antagonists & inhibitors ; Down-Regulation ; Guanidines ; pharmacology ; Humans ; Imidazoles ; pharmacology ; Interleukin-8 ; metabolism ; K562 Cells ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; Sulfones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

This study was aimed to investigate the influence of short hairpin RNA (shRNA) on proliferation of human leukemia cell line THP-1. The shRNA targeting the site 732-752 of DOT1L mRNA was designed and chemically synthesized, then a single-vector lentiviral, tet-inducible shRNA-DOT1L system (Plko-Tet-On) was generated. Thereafter, the THP-1 cells with lentivirus were infected to create stable cell line with regulatable shRNA expression. The expression of DOT1L in the THP-1 cell line was assayed by RT-PCR. Effect of shRNA-DOT1L on the proliferation of THP-1 cells was detected with MTT method,and the change of colony forming potential of THP-1 cells was analyzed by colony forming unit test. Cell cycle distribution was tested by flow cytometry. The results indicated that the expression of DOT1L was statistically lower than that in the control groups. The proliferation and colony forming capacity of THP-1 cells were significantly inhibited. The percentage of cells at G0/G1 phase increased in THP-1/shRNA cells treated with Dox while the percentage of cells at S phase significantly decreased as compared with that in the control group. It is concluded that the shRNA targeting DOT1L can effectively inhibit the proliferation of acute monocytic leukemia cell line THP-1.
Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Leukemia, Monocytic, Acute ; genetics ; Methyltransferases ; genetics ; RNA, Small Interfering