Objective To investigate the effects of different stabilizers, time and temperature before centrifugation on the stability of homocysteine (Hcy) and other related thiols levels involving EDTA-containing whole blood.Methods Blood was drawn from 17 healthy adults and collected into tubes containing EDTA, EDTA plus NaF and EDTA plus 3-deazaadenosine(3DA),then stored on crush ice(0-4 ℃) immediately or at room temperature(25 ℃).Plasma was separated at 0, 3, 6, 24 and 48 hours, respectively.The levels of plasma total Hcy (tHcy), total cysteine (tCys), tatal cysteinylglycine (tCysGly) and tatal glutathione (tGSH) were measured by HPLC.The plasma immediately separated was used as baseline sample. Results In EDTA tubes stored at room temperature, tHcy levels increased by 38.5%, 64.2%, 141.9%, 225.4% for 3, 6, 24, and 48 h, respectively.The levels of tCysGly and tGSH increased by 20.0% and 37.9% within 3 h, however, tCys decreased by 3.5%.The levels of the thiols increase by less than 5% up to 6 h in EDTA tubes stored on crush ice.In EDTA-3DA and EDTA-NaF tubes, no statistical differences were observed in the plasma levels of tHcy, tCys,tCysGly and tGSH compared with their respective baseline values at room temperature for 3 h(EDTA-3DA tubes:F=0.01,0.94,0.09,0.01,all P>0.05;EDTA-NaF tubes:F=0.85,0.04,0.03,0.02,all P>0.05).Conclusions The EDTA-plasma levels of tHcy and other related thiols are time and temperature-dependent. There is a strong need for standardization of blood sample collection and processing in tHcy and other thiols assays. The plasma concentrations of tHcy, tCys, tCysGly and tGSH were stable for 3 h at least in the EDTA-3DA and EDTA-NaF tubes kept at room temperature.

Objective:To analysis the effects of alprostadil and yishen huashi particles on blood glucose,blood lipid,renal function and urinary podocyte proteins of patients with diabetic nephropathy.Methods:98 patients with diabetic nephropathy were selected and randomly divided into the control group and the experimental group with 49 cases in each group.The patients in the control group were treated with alprostadil,while the patients in the experimental group were treated with yishenhuashui particles on the basis of the control group.Then the curative effect,the levels of glycosylated hemoglobin (HbAlc),blood glucose (FPG),2 h postprandial blood glucose (2h PG),triglycerides (TG),total cholesterol (TC),high-density lipoprotein (HDL-C),low density lipoprotein (LDL-C),blood urea nitrogen (BUN),creatinine (Cr),β2 microglobulin (β2-MG),bladder inhibition (Cys-C) and urinary podocyte proteins (PCX) in the two groups were observed and compared between the two groups before and after the treatment.Results:The total effective rate of experimental group was higher than control group (P＜0.05).After treatment,there was no statistically significant difference about the HbA1c,FPG and 2 HPG between the two groups (P＞0.05).After the treatment,the levels ofTG,TC,LDL-C,BUN,Cr,β2 MG,Cys C,PCX and urinary nephrin/urine Cr of the experimental group were lower than those of the control group (P＜0.05).The HDL-C of experimental group was higher than that of the control group (P＜0.05).Conclusion:The curative is effect of alprostadil and yishen huashi particles in treatment diabetic nephropathy patients,can conducive to the improvement of blood glucose,blood lipid,renal function,reduce the concentration of urinary podocyte related proteins.

Objective:To discuss the therapeutic effect of blood purification on hemorrhagic fever with renal syndrome combined with acute renal failure.Methods:60 patients with hemorrhagic fever with renal syndrome combined with acute renal failure were selected,and they were divided into two groups according to the odd and even number of hospitalization.The observation group (31 cases) received hemodialysis therapy.The control group (29 cases) received veno-venous hemofiltration therapy.The renal function,serum ion,blood routine indexes and the incidence of adverse reactions during treatment was detected and compared between two groups.Results:There were no statistical significance on K+,HCO3-,BUN,Cr between two groups (P＞0.05).After treatment,the K+,BUN,Cr levels were decreased in two groups (P＜0.05).The values of observation group were lower than those of the control group (P＜0.05).The level of HCO3-was increased in two groups,and it was higher in observation group than that of the control group (P＜0.05).Before treatment,there was no statistical significance on Hb,Hc,PLT between two groups (P＞0.05).After treatment,the Hb,Hc,PLT levels were increased in two groups (P＜0.05),but there were no statistical significance between two groups (P＞0.05).During 1 year of follow-up,the 6 months and 12 months survival rate of observation group was higher than that of the control group (P＜0.05).During treatment,there were no statistical significance on the incidence of adverse reactions between two groups (P＞0.05).Conclusions:The hemofiltration therapy has a good therapeutic effect on blood purification on hemorrhagic fever with renal syndrome combined with acute renal failure,and it can regulate the electrolyte balance,improve the renal function and increase the survival rate,which is equivalent to veno-venous hemofiltration therapy in amelioration of blood state.

AIM:To investigate the role of peripheral blood leukocytes and polymorphonuclear neutrophils (PMNs) on intestinal injury following mesenteric ischemia/reperfusion (IR) in rats. METHODS: Twenty adult, male Sprague-Dawley rats, weighing 200-230 g, were randomly divided into two groups. The control group (CON) consisting of 10 rats was subjected to laparotomy and separation of superior mesenteric artery (SMA) only. The ischemia/reperfusion (IR) group consisting of 10 rats, was subjected to laparotomy, followed by occlusion of the superior mesenteric artery (SMA) by an atraumatic microvascular clamp for 30 min. At the end of ischemic period in IR, the microvascular clamp was removed and the intestinal segment was reperfused for 60 min. The pathological changes of the ileal mucosal tissue were evaluated. The apoptosis of intestinal mucosal epithelial cells was examined by terminal deoxylnucleotidy-l transferase mediated-dUTP nick end-labeling (TUNEL). The enzymatic activity of casapse-3 in mucosal cells was determined using a colorimetric assay. The percentages of apoptotic peripheral blood leukocytes and PMNs were measured by flow cytometry using Annexin-V/PI double staining assay. The numbers of peripheral blood leukocytes in each animal was measured at baseline, 30 min of ischemia, and 30 min and 60 min of reperfusion. RESULTS: (1) Compared to CON group animals, the most severe mucosal injury was observed in IR group under optical microscope. (2) The number of apoptotic mucosal epithelia cells and enzymatic activity of caspase-3 were significantly higher in IR than those in CON group (P

A 33-year-old male patient with tick-borne encephalitis (TBE) was reviewed, who presented with severe neurological deficits following TBEV infection, and improved in his motor and quality of life after an individualized rehabilitation.

Objective To investigate the tryptophan (Trp) metabolism change in the patients with chronic hepatitis B (CHB) and hepatitis B cirrhosis (HBC) and to analyze the related risk .Methods Eighty-one cases of CHB and 40 cases of HBC hospitalized in the First Affiliated Hospital of Chongqing Medical University from October 2015 to January 2016 served as the stud-y subjects ,and 40 healthy persons as the controls .High performance liquid chromatography -fluorescence detection (HPLC-FD) were adopted to detect the levels of plasma Trp ,kynurenine (Kyp) and its metabolite 5-hydroxy tryptamine (5-HT) .HBV-DNA vi-ral replication number was detected by real-time quantitative PCR .The serological indicators differences among 3 groups were ana-lyzed .The risk factors were analyzed and screened by the Logistic regression analysis and Pearson correlation analysis .Results Compared with the control group ,plasma 5-HT and Trp levels in the CHB group and HBC group were significantly decreased (P<0 .01);plasma Kyn/Trp in the HBC group was significantly higher than that in the other two groups (P<0 .01) .Kyn/Trp in the HBC group was negatively correlated with A/G (r= -0 .686 ,P<0 .01) ,and Kyn/Trp in the CHB group was positively correlated with Log DNA (r= 0 .784 ,P<0 .01) .The Logistic regression analysis indicated that Kyn/Trp was the risk factor of CHB and HBC .Conclusion Detecting plasma Trp and its metabolite can serve as the adjuvant indicators for assessing the progression of hep-atitis B .

Objective To observe the expression of NOD2 and epithelial-mesenchymal transition (EMT) related proteins in podocytes in high glucose environment,and explore the molecular mechanism of NOD2 involved in EMT.Methods The human glomerular podocytes were the subjects of study.α-SMA and Nephrin expressions were detected by immunofluorescence;the mRNA and protein expressions of NOD2,Snail and EMT related proteins (α-SMA,Desmin,E-cadherin,Nephrin) were detected by real-time fluorescence quantitative PCR and Western blotting.The podocytes were stimulated by high-glucose after shRNA interfering the of NOD2 expression,and the expressions of Snail and subsequent EMT-related proteins were detected by Western blotting.Prior to the activation of NOD2 by muramyl dipeptide (MDP),shRNA was used to interfere with the expression of Snail.E-cadherin,Nephrin,Desmin,and α-SMA were detected by Western blotting.Results After 24 hours of high glucose stimulation,PCR and Western blotting results showed that the expressions of NOD2 and Snail were significantly increased;the expressions of epithelial phenotype proteins E-cadherin and Nephrin were down-regulated;and the expressions of interstitial phenotype proteins Desmin and α-SMA were increased (all P ＜ 0.05);while there was no significant change in the hypertonic control group.After interference with NOD2,the abnormal expression of Snail and EMT related proteins were all recovered.After interference with Snail expression,Compared with the MDP group,the protein expressions of E-cadherin and Nephrin were significantly increased (all P ＜ 0.05);the expressions of Desmin and α-SMA were significantly decreased.Conclusions High glucose can induce NOD2 expression in podocytes,and promote podocyte epithelial-mesenchymal transition by upregulating Snail expression.Gene intervention targeting the NOD2/Snail/EMT pathway can reduce high-glucose-induced podocyte injury and may provide new ideas for the treatment of diabetic nephropathy.

Objective:To explore the expression of fructose bisphosphate aldolase A (ALDOA) in the bone marrow of patients with acute myeloid leukemia (AML) and the correlation with clinical features and prognosis.Methods:The bone marrow samples of 90 newly diagnosed AML (non-acute promyelocytic leukemia) patients and 18 allogeneic hematopoietic stem cell transplantation donors who were treated from January 2013 to December 2015 in the First Affiliated Hospital of Zhengzhou University and the Children's Hospital Affiliated to Zhengzhou University were collected. The relative expression level of ALDOA mRNA in bone marrow samples was detected by using real-time quantitative polymerase chain reaction (qRT-PCR). Clinical data of these patients were retrospectively analyzed, and the patients were divided into continuous complete remission (CR) group and refractory recurrent (RR) group according to the clinical response and follow-up results. The differences of the relative expression level of ALDOA mRNA between AML group and the normal control group, CR group and RR group were analyzed. Univariate and multivariate Cox regression risk model were used for analysis of factors influencing prognosis of AML patients.Results:The relative expression level of ALDOA mRNA in AML group was higher than that in normal control group [(5.71±0.44) vs. (1.10±0.08), t = 4.74, P<0.001]. The relative expression level of ALDOA mRNA in the RR group was higher than that in the CR group [(6.69±0.67) vs. (4.30±0.36) , t = 2.79, P < 0.001]. In addition, there were statistically significant differences in the proportion of patients with ALDOA mRNA high expression and those with ALDOA mRNA low expression stratified by the number of white blood cell, the proportion of bone marrow blasts and whether complete remission could be achieved or not after 1 course of induction therapy (all P < 0.05). Overall survival in patients with ALDOA high expression was worse than that in patients with ALDOA low expression ( χ2 = 5.59, P = 0.018). Multivariate analysis showed that white blood cell count, prognosis stratification, whether complete remission could be achieved or not after 1 course of induction therapy and ALDOA expression were the independent prognostic factors for the death of AML patients (all P < 0.05). Conclusions:ALDOA may play an important role in the development and progression of AML, and the expression level of ALDOA in the bone marrow can be used as an index for the prognosis assessment of AML patients and may be a potential therapeutic target for AML.

Objective:To investigate the effects of gestational diabetes mellitus (GDM) on neonatal metabolites.Methods:This retrospective cohort study recruited 580 singleton newborns who were born to women with GDM from January 2018 to December 2018 in Foshan Maternal and Child Health Care Hospital as the GDM group. Another 580 counterparts born to non-GDM singleton mothers with matching age were selected as the non-GDM group with an allocation ratio of 1 to 1. Neonatal genetic metabolic disease screening was performed within 3-7 days after birth. Two independent sample t-test, and multiple linear regression model were used for statistical analysis. Results:There were significant differences in seven amino acids and 10 fatty acids levels between the GDM and non-GDM group. The serum levels of six amino acids and eight fatty acids were increased in the GDM group, while the levels of piperamide [(140.79±32.60) vs (150.26±35.46) μmol/L, t=－4.733, P<0.001], palmitoyl carnitine [(2.59±0.81) vs (2.73±0.82) μmol/L, t=－2.940, P=0.003], and carbamate [(0.066±0.022) vs (0.069±0.022) μmol/L, t=－1.937, P=0.042] were decreased compared with the non-GDM group. After adjusting for maternal gravidity, parity, neonatal birth weight, and gender, multivariate linear regression analysis showed that GDM was positively correlated with three amino acids levels, which were cysteine ( ?=0.012), homocysteine ( ?=0.263) and leucine ( ?=4.225); and was negatively correlated with glycine ( ?=－6.271) and piperamide level ( ?=－9.885). With regard to the fatty acids, GDM was positively correlated with the neonatal propionyl carnitine ( ?=0.214), butyryl carnitine ( ?=0.014), 3-hydroxybutyryl carnitine ( ?=0.006), isovaleryl carnitine ( ?=0.009), 3-hydroxyisovaleryl carnitine ( ?=0.024), hexadecanoyl carnitine ( ?=0.001), decadienoyl carnitine ( ?=0.045), octadecadienyl carnitine level ( ?=0.128), but was negatively correlated with palmitoyl carnitine ( ?=－0.119), and carbamate ( ?=－0.002) (all P<0.05). Conclusions:Correlations between maternal GDM and the levels of amino acids and fatty acids in neonates was noted in this study, suggesting that maternal GDM may affect the metabolism of amino acids and fatty acids in offspring at early stage of life.

Objective:To analyze the clinical data of pregnant women complicated with cardiovascular disease in our center in the past 10 years, and to explore the trend of incidence, clinical diagnosis, and treatment of the disease.Methods:Clinical data of pregnant women with cardiovascular disease who delivered in Beijing Anzhen Hospital, Capital Medical University from 2010 to 2019 were collected and analyzed retrospectively. According to the time of the establishment of multidisciplinary team (MDT) in the center, the pregnant women were divided into the first 5-year group (2010-2014) and the second 5-year group (2015-2019). The general data, the composition of pregnancy complicated with cardiovascular disease and the changes of maternal and infant outcomes of the two groups were analyzed.Results:(1) During 2010-2019, there were 2 267 cases of pregnancy complicated with cardiovascular disease (836 cases in the first 5-year group and 1 431 cases in the second 5-year group), with a total incidence of 10.2% (2 267/22 334). Among all kinds of cardiovascular diseases, arrhythmia (41.0%, 930/2 267) and congenital heart disease (38.2%, 865/2 267) were more common. (2) There were 212 cases (25.4%, 212/836) and 426 cases (29.8%, 426/1 431) classified as Ⅲ or Ⅳ by modified WHO cardiovascular risk classification in the first 5-year group and the second 5-year group, respectively, and the difference was statistically significant ( χ2 =5.076, P=0.024). Among all kinds of cardiovascular diseases, there were 111 cases (13.3%, 111/836) and 159 cases (11.1%, 159/1 431) with valvular disease in the first 5-year group and the second 5-year group, respectively. The change of the component ratio was -16.5% (the difference was significant when the absolute value of change>10%), showing a significant decreasing trend. Aortic disease was found in 16 cases (1.9%, 16/836) and 56 cases (3.9%, 56/1 431), respectively, with a significant upward trend of 105.3%. (3) The mortality rate of pregnant women with cardiovascular disease was 1.0% (22/2 267), and 1.2% (10/836) and 0.8% (12/1 431) in the first 5-year grouop and the second 5-year group, respectively. There was no significant difference between the two groups ( χ2=0.702, P=0.402). ICU occupancy rates in the first 5-year group and the second 5-year group were 25.6% (214/836) and 20.7% (296/1 431), respectively, and the difference between the two groups was statistically significant ( χ2=7.306, P=0.007). There were no significant differences in cesarean section rate, mortality rate and incidence of adverse events between the two groups of pregnant women, and there were no significant differences in birth weight, preterm birth rate, mortality rate and asphyxia rate between the two groups of newborns (all P>0.05). Conclusions:Pregnancy complicated with cardiovascular disease is a common cause of adverse obstetric outcomes. There are various types of specific cardiovascular diseases, and the prognosis varies greatly. In recent years, the disease composition ratio has changed, and the severity and complexity of diseases have increased. Hierarchical management, MDT and individual management could improve the treatment level and reduce adverse outcomes.