A rapid method has been developed based on the sample preparation procedure named as QuEChERS (Quick,Easy,Cheap,Effective,Rugged and Safe),combined with reversed-phase high performance liquid chromatography with fluorescence detector and C18 column after precolumn derivatization using o-phthalaldehyde and 2-mercaptoethanol to determine dopamine in porcine muscle.Methanol and deionized water (0.1％ acetic acid,v/v) with a ratio of 60∶40 was used as mobile phase.The flow rate was 0.8 mL/min and dopamine was eluted within 15 min.The linearity range was 0.003-8 μg/mL with r=0.9992.The detection limit for dopamine was 4 μg/kg and the quantification limit was 9 μg/kg.Recovery studies were carried out at 0.1,0.5 and 1.0 mg/kg fortification levels and the average recoveries obtained ranged from 90.4％ to 98.2％ with relative standard deviations between 3.5％ and 8.1％.The method was found to be suitable for detection of dopamine in animal product tissues at the maximum residue level.

Air Pollutants, Occupational ; analysis ; Ammonium Sulfate ; analysis ; Chromatography, Ion Exchange ; methods ; Workplace

Objective To explore the occurrence of anemia, anaemia type, iron metabolism situation,variation of serum Prealbumin (PA), and the relationship among them in cancer patients. Methods Three hundred and seventy cancer patients, admitted from July 2007 to April 2010 to our hospital, were enrolled into the study. The clinical data of all subjects were analyzed retrospectively. Results The incidence of anemia in 370 patients was 55.67% (206/370). Among all 206 anemia cases,129 cases had mild anaemia and 77 cases had middle to severe anaemia. In the last group ( n = 77 ), we found significant decrease in serum iron level ( [ 8. 37 ± 6. 09 ] μmol/L) and increase in serum ferritin level ( [ 474. 57 ± 327. 58 ] μg/L); and the correlation between serum iron and the anemia degree( Ps ＜ 0. 05 ). However, we found no significant differences of serum ferritin level between the groups with different degree of anemia(P ＞0. 05). Among all 206 anemia cases ,187(90. 77% ) patients had a low level of serum PA, but no relationship between the degree of anemia and the drop of serum PA ( P ＞ 0. 05 ). Conclusion The anemia was very popular in cancer patients, which had correlation with iron metabolism situation but not PA.

A rapid method has been developed based on the sample preparation procedure named as QuEChERS （Quick, Easy, Cheap, Effective, Rugged and Safe）, combined with reversed-phase high performance liquid chromatography with fluorescence detector and C18 column after precolumn derivatization using o-phthalaldehyde and 2-mercaptoethanol to determine dopamine in porcine muscle. Methanol and deionized water （0.1% acetic acid, v/v） with a ratio of 60：40 was used as mobile phase. The flow rate was 0.8 mL/min and dopamine was eluted within 15 min. The linearity range was 0.003-8 μg/mL with r=0.9992. The detection limit for dopamine was 4 μg/kg and the quantification limit was 9 μg/kg. Recovery studies were carried out at 0.1, 0.5 and 1.0 mg/kg fortification levels and the average recoveries obtained ranged from 90.4% to 98.2% with relative standard deviations between 3.5% and 8.1%. The method was found to be suitable for detection of dopamine in animal product tissues at the maximum residue level.

To investigate the role of the extracellular signal-regulated kinase (ERK1/2) and PI3K/AKT/ mTOR signal pathway inducing bone marrow mesenchymal stem cells (BMSCs) differentiation into neural cells, mouse bone marrow-derived mesenchymal stem cell lines D1 cells were used as research object. And they were divided into control groups and salidroside (SD) groups. Different concentrations (5, 25, 50, 100 and 200 microg x mL(-1) of SD were used and SD (100 microg x mL(-1)) was used to induce at different time (0.5, 1, 3, 6, 9, 12, 24, 48 and 72 h). The immunofluorescence staining chemical technology, real-time PCR and Western blotting were used to detect the positive rates of NSE, MAP2, beta-Tubulin III, NES, GFAP and the expression levels of beta-Tubulin III, NSE, ERK1/2, AKT. The expression of ERK1/2 and NSE was detected when the ERK1/2 and PI3K/AKT/ mTOR signal pathway was blocked by PD98059 and LY294002. It indicated that the positive rates of NSE, MAP2, beta-Tubulin III, NES and GFAP were gradually enhanced with time increased. The expression level of NSE and beta-Tubulin III protein were significantly higher than those in control groups (P < 0.01). The expression of ERK1/2, AKT mRNA and protein were higher with concentration and time increased. When the ERK1/2 and PI3K/AKT/mTOR signal pathway were blocked, the expression levels of NSE, NES and beta-Tubulin III mRNA and NSE protein were inhibited significantly. It points out that SD can stimulate the ERK1/2 and PI3K/AKT/mTOR signal pathway to promote BMSCs differentiation into neural cells.

Objective To investigate the protective effect of icariin against varicocele-induced damage on rat epididymis. Methods Forty adolescent male SD rats were randomly divided into control group (n=10), experimental varicocele (EV) group (n=15), icariin (ICA) therapy group (n=15). Experimental varicocele model in the EV group and ICA group was established. The EV was induced by partial ligation of the left renal vein. The rats in the control group underwent a sham operation that separated the spermatic vessels without ligation. Each rat in the control group and EV group was lavaged with 2 mL physiological saline every day for 6 weeks. Each rat in the ICA group was lavaged with icariin [100 mg/(kg·d)] for 6 weeks. Rats in all groups were executed after 6 weeks. The contents of sialic acid were measured by spectrophotometry. Carnitine concentrations were measured by DTNB. HE stain was used to observe the microstructure changes in the epididymal tissue. Electron microscopy was used for observing the ultrastructural changes of the epididymis. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method was used to detect the apoptosis of the epididymal epithelium. Results Compared with the control group, the microstructure and ultrastructure of the epididymis in EV group showed pathological damage. Compared with the EV group, the damage of the epididymal microstructure and ultrastructure significantly alleviated. Apoptosis index (AI) of epididymal epithelium in the EV group was significantly higher than that in the control group (P<0.01). However, AI of epididymal epithelium in the ICA group was significantly lower than that in the EV group (P < 0.01). The sialic acid and carnitine concentrations of the epididymis in the EV group was significantly lower than that in the control group (P < 0.01), respectively. However, the sialic acid and carnitine concentrations of the epididymis in the ICA group was significantly higher than that in the EV group (P < 0.01), respectively. Conclusion This study indicates that varicocele could result in the apoptosis of epididymal epithelium and icariin decreased the varicocele-induced apoptosis , suggesting that varicocele could damage the structure and function of epididymis, which can be repaired by icariin.

Objective:To illustrate a relatively complete knowledge system (e.g., research outputs, current hotspots, and future trends) in the sepsis field and to help scholars grasp the scientific research direction or clinical focus of treatment.Methods:The relevant literatures of sepsis during the time from 1985 to 2019 in Web of Science database were collected. Sepsis-related research contents were generated using softwares (CiteSpace 5.6.R2 and VOSviewer 1.6.13), which using data mining, information processing and knowledge map methods, to analyze the historical evolution and predict the development trend.Results:A total of 8 189 papers on sepsis were published. The volume of publications were increasing yearly from 1985 to 2019, and reached the top list of 1 276 in 2019. For research contents of sepsis, it has formed the basic characteristics of sepsis which focusing on epidemiological studies and animal experiments. Through cluster analysis, the researches mainly focused on six aspects: septic rat, necrotizingenterocolitis, sepsis-associated encephalopathy, acute kidney injury (AKI), gut-derived sepsis, and inflammatory mediator. And it presented the literature characteristics that related to the injury or dysfunction of intestines, brain, liver, kidney or other organs, but the heart and lung researches were more marginal. Additionally, based on the top key words with the strongest citation bursts, it reflected that the development trend of the continuous attention hotspots with "endotoxin" or "endotoxin shock", the significant attention hotspots with "inflammation", "immunity" and "multiple organ dysfunction syndrome" (MODS), and the novel burst attention hotspots with sepsis management including "diagnosis" and "chemotherapy".Conclusions:Through the hotspots and trends visualization of sepsis, the current researches are prefer to animal experiments, epidemiology, or other basic scientific aspects. Meanwhile, the researches are mostly focusing on inflammatory reaction, immune function or organ dysfunctions. Integrating the knowledge maps of hotspots and trends, based on researches of epidemiology, diagnosis, risk factors, pathogenesis, or treatment, we predict that the future scientific topics will concentrating on childhood sepsis, organ injury mechanism or intervention relating to MODS, and integrated management of sepsis by combining traditional Chinese medicine and Western medicine.

Objective:To investigate the effect of astragaloside IV (AS-IV) on the secretion of stromal cell-derived factor-1α (SDF-1α) and CXC chemokine receptor 4 (CXCR4) by high glucose injured human umbilical vein endothelial cells (HUVECs), so as to lay a foundation for further study on AS-IV improving angiogenesis by regulating SDF-1 α/CXCR4 axis of endothelial cells.Methods:HUVECs were isolated and cultured from the umbilical vein of full-term healthy newborns and identified by von Willebrand factor (vWF) combined with 4-diamino-2-phenylindole (DAPI) nuclear staining. The obtained HUVECs was cultured in EGM-2 medium with 30 mmol/L glucose for 120 h to obtain high glucose damaged HUVECs. After intervention with different concentration gradients (25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) AS-IV for 72 hours, the contents of SDF-1α and CXCR4 were detected by enzyme linked immunosorbent assay (ELISA) method to determine the best concentration of AS-IV. The supernatant of damaged HUVECs were collected at 6, 12, 24, 48 and 72 hours after intervention with the best concentration of AS-IV, and the contents of SDF-1α and CXCR4 were detected by ELISA method to determine the best action time of AS-IV. The damaged HUVECs was randomly divided into experimental group and control group, and the blank group was set up at the same time. The experimental group was treated with the best concentration of AS-IV and the best time, the control group and the blank group were treated with the same volume of phosphate buffered saline (PBS) solution, and the contents of SDF-1α and CXCR4 in each group were detected by ELISA method.Results:The vWF factor on the cell membrane was green fluorescence, and the nucleus was blue after DAPI staining. When the fusion image showed green fluorescence, HUVECs were identified by blue fluorescence. The expression of SDF-1α in damaged HUVECs was the best when treated with AS-IV of 100 mg/L for 24 hours (1 642.87 pg/ml), and the expression of CXCR4 in damaged HUVECs was the best when treated with AS-IV of 50 mg/L for 48 hours (8.44 ng/ml). Compared with the control group, the contents of SDF-1α and CXCR4 in the experimental group were significantly increased, and the difference was statistically significant ( P<0.05). While the contents of SDF-1α and CXCR4 in the experiment group were slightly less than those in the blank group and there was no statistically significant difference ( P>0.05). Conclusions:AS-IV can promote the expression of SDF-1α and CXCR4 in HUVECs damaged by high glucose to return to normal physiological level, so as to play the role of vascular repair and neovascularization.

Objective To study the mRNA expression of HS1-associated protein X-1(Hax-1),an an- ti-apoptosis genc,in the peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythe- matosus(SLE),and further investigate the roles and significance of Hax-1 in the pathogenesis of SLE.Meth- ods Generation of longer cDNA fragments from serial analysis of gene expression(SAGE)tags for gene identi- fication(GLGI)was applied to identify the gene Hax-1 according to the Long SAGE tag.Then reverse tran- scription-polymerase chain reaction(RT-PCR)technique was used to semiquantitatively analyze mRNA ex- pressions of Hax-1 in PBMC from 34 active SLE patients and 25 healthy subjects.Results Compared with healthy controls,there was significant difference between SLE patients in the active stage and the normal controls(Z=-4.556,P＜0.01).The average level of mRNA expression in active SLE group was higher than that in healthy controls.Significant difference was found between the group with mild SLE and either the moderate or the severe one(P＜0.01).Conclusion The mRNA expression level of Hax-1 in active SLE group increase markedly,and to some extent,it is related to the activity of SLE.This provides a valuable basis for the further study on the role of apoptosis in SLE.

Human ScFv against botulinum neurotoxin serotype A(BoNTa) was modified by fusing human IgG1 Fc to C terminal of ScFv. ScFv-Fc fusion protein was expressed at high level over 30% of total host cell proteins in E.coli. Recombinant protein existed in inclusion body form. Renatured ScFv-Fc was purified to 90%~95% by Protein G Sepharose column. In vitro ScFv-Fc could bind specific to toxiod BoNTa in ELISA. Recombinant ScFv-Fc had similar relative affinity to parent ScFv and had improved stability.