Determination of sodium valproate and vancomycin in human serum by HPLC-MS/MS / 药学实践杂志

Objective To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneous determination of sodium valproate and vancomycin in human serum. Methods Valproic acid-d6 and kanamycin B were used as the internal standard of sodium valproate and vancomycin, the serum samples were treated by acetonitrile precipitation protein method. The mobile phase was 0.1% formic acid aqueous solution-acetonitrile for gradient elution. The flow rate was 0.5 ml/min, with column temperature at 25 ℃. The sample volume was 4 μl and total analysis time was 12 min. The positive and negative ion mode was monitored by electrospray ion source and the multiple reaction monitoring mode was used for quantitative analysis. The specificity, standard curve, lower limit of quantification, precision, recovery, matrix effect, and stability of the method were examined. Results Sodium valproate and vancomycin had good linear relationships in the range of 1 - 200 μg/ml and 0.5 - 100 μg/ml, respectively. The quantitative lower limits were 1 μg/ml and 0.5 μg/ml, respectively. The extraction recoveries were above 70%. The inter- and intra-batch precision RSD values were less than 10%. The stability was good and there was no obvious matrix effect. Conclusion This method is simple, quick, sensitive, specific and accurate, which could be used to simultaneously determine the concentration of sodium valproate and vancomycin in human serum.

Exploration and practice on the opening and sharing management mode of undergraduate experimental teaching instruments and laboratory information management system (LIMS) under the background of "Double first-class" / 生物工程学报

In order to explore the role of the opening and sharing management mode (OSMM) of undergraduate experiment teaching instruments in fostering talents under the "Double first-class" initiative, the importance of laboratory information management system (LIMS) in the OSMM was discussed, in light of the experience and shortcomings of developing an OSMM in the experimental teaching center of environment and ecology college at Xiamen university. Some approaches were put forward to promote OSMM development and improve the utility of teaching equipments, so as to maximize the utility of the instruments, guarantee the regular teaching and scientific research, promote the sustainable and healthy development of colleges and universities, and achieve the desirable progress of the ongoing national "Double first-class" initiative.

Study on the synergistic effects of aspirin and atorvastatin on cell proliferation of non-small cell lung cancer cells / 药学实践杂志

Objective To study the synergistic effects of aspirin and atorvastatin on cell proliferation of non-small cell lung cancer cell A549 and NCI-H460 and the mechanism of these actions. Methods The proliferation of A549 and NCI-H460 cells treated by aspirin or/and atorvastatin were determined by MTS assay. The migration of A549 and NCI-H460 cells were conducted by wound-healing assay. The expression of relevant protein in mTOR and NFκB signaling pathway were detected by western blotting. The mRNA expression of TNF-α and IL-1β were detected by quantitative real-time PCR. Results Aspirin or/and atorvastatin inhibited the proliferation and migration of A549 and NCI-H460 at concentration of 100 and 5 μmol/L or greater. The effect was enhanced by the combination of aspirin and atorvastatin. Aspirin or/and atorvastatin inhibited the protein expression of the phosphorylation of mTOR and NFκB, and down-regulated anti-apoptotic regulators Bcl-2 and Mcl-1 in NCI-H460 cells. The combination treatment of aspirin and atorvastatin was more efficacious than the single treatment. Atorvastatin decreased the mRNA expression of TNF-α. The combination of atorvastatin with aspirin decreased the mRNA expression of IL-1β by nearly 50 percent compared to the control (P<0.05). Conclusion Aspirin and atorvastatin have synergistic inhibitory effects on cell growth of non-small cell lung cancer cell A549 and NCI-H460 by suppressing mTOR and NFκB signaling pathway.

Management pattern of undergraduate teaching laboratory with a focus on creating the educational environment / 生物工程学报

The teaching laboratory is the central place to teach lab courses to undergraduate students. It is thus critical to evaluate how lab skills of students will be affected by the management of this lab. This study attempts to introduce the 6S concept, referring to "Seiri", "Seiton", "Seiso", "Seiketsu", "Shisuke", and "Safety", to the management of teaching laboratory, in particular, to the management of environments, safety, item deposition, reagents and consumables in the laboratory. We find that applying the "6S" management maintains a tidy lab environment, facilitates teaching the lab skills, and improves the quality of teaching, which together achieves the goal of education with the atmosphere. This study can provide a guideline for establishing and managing teaching laboratories in universities.

Expressions of Th17 lymphocytes and interleukin-17 in peripheral blood of patients with non-small cell lung cancer and their clinical significances / 肿瘤研究与临床

Objective To investigate the expressions of Th17 lymphocytes and interleukin-17 (IL-17) in peripheral blood of patients with non-small cell lung cancer (NSCLC) and its clinical significance. Methods Sixty patients with primary and untreated NSCLC were enrolled and designed as experimental group, at the same time, 60 healthy volunteers were collected as control group. Flow cytometry (FCM) was used to detect the level of Th17 lymphocytes. Enzyme linked immunosorbent assay (ELISA) was used for detecting the level of IL-17. The relationship between the expression levels of Th17 and IL-17 in peripheral blood and clinicopathological features was compared between the two groups. Results The peripheral blood levels of Th17 lymphocytes and IL-17 in the experimental group [(1.7±1.2) ％, (8.3±2.5) pg/ml] were higher than those in the control group [(0.9 ±0.6) ％, (5.4 ±1.2) pg/ml] (P< 0.05). The peripheral blood expression of Th17 lymphocytes and IL-17 in patients with smoking history [(1.8±1.2) ％, (8.8±3.7) pg/ml] were higher than those in patients without smoking history [(1.6±1.2)％, (8.0±2.2) pg/ml], and the peripheral blood expression of Th17 lymphocytes and IL-17 were higher in patients with squamous-cell carcinoma [(1.8 ±1.2) ％, (9.4 ±4.7) pg/ml] than those in patients with adenocarcinoma [(1.6±1.1) ％, (7.3±3.9) pg/ml], furthermore, they were also higher in patients with stage Ⅲ-Ⅳ than those in patients with stage Ⅰ-Ⅱ (P < 0.05). Conclusion Th17 lymphocytes and IL-17 play certain roles in the occurrence and progression of NSCLC.

Onco-miR-24 regulates cell growth and apoptosis by targeting BCL2L11 in gastric cancer

Gastric cancer is one of the most common malignancies worldwide; however, the molecular mechanism in tumorigenesis still needs exploration. BCL2L11 belongs to the BCL-2 family, and acts as a central regulator of the intrinsic apoptotic cascade and mediates cell apoptosis. Although miRNAs have been reported to be involved in each stage of cancer development, the role of miR-24 in GC has not been reported yet. In the present study, miR-24 was found to be up-regulated while the expression of BCL2L11 was inhibited in tumor tissues of GC. Studies from both in vitro and in vivo shown that miR-24 regulates BCL2L11 expression by directly binding with 3'UTR of mRNA, thus promoting cell growth, migration while inhibiting cell apoptosis. Therefore, miR-24 is a novel onco-miRNA that can be potential drug targets for future clinical use.

miR-143 suppresses the proliferation and migration of SGC7901 gastric cancer cells / 中国肿瘤临床

Objective:To investigate the role and mechanism of miR-143 in the proliferation and migration of gastric cancer (GC) cells. Methods:Western blot was performed to detect the expression level of avian erythroblastosis oncogene B-3 (ERBB3) in GC tissues, paired non-cancerous tissues, and SGC7901 GC cells. RT-qPCR was conducted to determine the mRNA and miR-143 of ERBB3 quantita-tively. Bioinformatics tools were used to predict the target gene of miR-143. Luciferase reporter assay was carried out to confirm the predicted target gene. Transwell and EdU assays were applied to observe the migration and proliferation of SGC7901 GC cells transfect-ed with miR-143 mimics/inhibitor/NC mimics/inhibitor. Results:Compared with the expression levels of ERBB3 and miR-143 in the paired non-cancerous tissues, the expression level of ERBB3 was upregulated and the expression level of miR-143 was downregulated in GC tissues. In the prediction of the potential target gene, miR-143 could bind to a specific sequence of the 3′-untranslated regions (UTR) of the mRNA of ERBB3. This finding was supported by luciferase reporter assay results. In vitro, ERBB3 protein expression and cell migration and proliferation were suppressed significantly in the SGC7901 cells transfected with miR-143 mimics. By contrast, these processes were remarkably enhanced when the cells were transfected with miR-143 inhibitor. Conclusion:miR-143 can suppress the migration and proliferation of GC cells by downregulating the expression of ERBB3.

Etiological agents distribution and epidemiology of viral diarrhea in children below 5 years old in He′nan ;province,2008-2015 / 中华传染病杂志

Objective To investigate the infectious status,etiological spectrum and epidemiological characteristics of rotavirus (group A/B/C),calicivirus (novovirus Ⅰ/Ⅱ,sapovirus),astrovirus and enteric adenovirus in diarrhea cases below 5 years old from 2008 to 2015 in Henan provinces.Methods Totally 2541 stool samples were collected from cases below 5 years old in four sentinel hospitals.All stool specimens were tested for group A rotavirus by double antibody sandwich ELISA method.G/P genotyping of group A rotavirus was determined by nested multiplex PCR.Viral RNA was extracted from all samples and rotavirus (group B/C),calicivirus,astrovirus and enteric adenovirus were detected by two-step reverse transcription-polymerase chain reation (RT-PCR)/PCR.Results One thousand four hundred and twenty-one out of 2 541 samples were positive with a total positive rate of 55 .9%,among which,102 were mixed infection.The isolation rate of rotavirus was 36.0% (914 samples)(group A:785 cases,group B:36 cases,group C:93 cases),calicivirus was 12.1 % (308 samples)(novovirus Ⅰ:64 cases,novovirusⅡ:193 cases,sapovirus:51 cases),astrovirus was 5 .9% (151 samples),enteric adenovirus was 1 .9%(48 samples).The group A rotavirus gene type combinations were composed mainly of G9P[8],G2P[4], G3P[8 ],G1P [8 ]and most cases were identified from September to November and March to May. Novovirus Ⅱ was predominant in calicivirus and most cases were identifed between March and May. Rotavirus or calicivirus infection was mainly among children aged 4—12 months or 3—5 years, respectively.Clinical manifestations included fever,diarrhea,vomiting,dehydration.Gender and region distributions differed according to the types of pathogen.Conclusions Group A rotavirus and novovirus Ⅱare the major viral pathogen in diarrhea cases younger than 5 years old in Henan province.Different viral infections exhibit extinct epidemiologic and clinical characteristics.

The application of time to positivity in blood culture / 中华检验医学杂志

Time to positivity(TTP) is a new parameter in blood culture field.The article shows us the concepts of TTP,differential time to positivity (DTTP),and introduces their relation with bloodstream infection (BSI),catheter-related bloodstream infection (CRBSI).Besides,it stresses TTP' s clinical application,including determining the severity of disease;identifying the isolates whether pollution or not; identification of isolate strains ; detection of the drug resistance of isolates ; evaluating the effect of antibiotic ; helping to adjust the therapeutic drug; diagnosing or excluding CRBSI by means of DTTP;deciding whether the catheter is the source of infection in patients with candidemia; understanding the epidemiological distribution of strain.At the same time,the article also describes the shortcomings and domestic current status.

Influence of arsenic trioxide in vasculogenic mimicry of HepG2 cells and its mechanism / 吉林大学学报(医学版)

Objective To investigate the influence of arsenic trioxide (AS2 O3 )in the vasculogenic mimicry (VM ) of HepG2 cells, and to preliminary clarify the possible mechanism of inhibition of AS2 O3 on the VM. Methods Themean inhibitory concentration (IC50 )of AS2 O3 72 h after treatment of HepG2 cells was calculated by CCK-8 assay.The HepG2 cells were cultured on 3-D Matrigel and randomly divided into control group, 1/2 IC50 AS2 O3 group and IC50 AS2 O3 group.IPP software was used to calculate the number,length and area of VM,and the expression levels of VM-related proteins VE-cadherin and MMP-2, apoptotic-related protein caspase-3 and proliferation-related protein PCNA were detected by Western blotting method.Results The IC50 of AS2 O3 was 10μmol·L-1 72 h after treatment of HepG2 cells.The number,length and area of VM in 1/2 IC50 and IC50 AS2 O3 groups were significantly lower than those in control group (P0.05).Conclusion AS2 O3 can inhibit the forming of VM of HepG2 cells,which indicated that its mechanism may be related to inhibiting the expressions of VE-cadherin and MMP-2 .

Quantitative analysis of lineage-specific chimerism using real-time PCR and single nucleotide polymorphisms / 白血病·淋巴瘤

Objective To develop a real-time quantitative PCR method with TaqMan probe to analysis the lineage-specific chimerism based on single nucleotide polymorphisms (SNP).Methods CD3 positive and CD15 positive cells were separated by magnetic cell sorting system from cord blood,and a quantitative method were established using real-time quantitative PCR and SNP.Detect the artificial chimerism and mark the standard curves.The reaction system was optimized,and the sensitivity and specificity were evaluated.Results Separation purity of blood cells by magnetic cell sorting system was up to 94 ％-97 ％.Discrimination between donor and recipient was possible.Dilution experiments of the mock chimerism sample revealed that Ct values correlated linearly with the logarithm of recipient/donor DNA fraction (r ＞ 0.98),and the limit of detection for a minor DNA percentage was under 0.1％,and the specificity was also good.Quantitative analysis of 4 clinical cases in same period were made by real-time fluorescence quantitative SNP-PCR,the fitted rate was 87.50 ％ based on the chimera standard curve calculated for 1 case.Conclusion The method shows high sensitivity and specificity,and will be used to quantify the lineagespecific chimerism after allogeneic hematopoietic stem cell transplantation.

Application of sorting peripheral blood cells by flow cytometry in the detection of peripheral Mood cell sorting / 白血病·淋巴瘤

Objective To apply fluo-rescencc-activated cell sorting(FACS) in sorting T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+), which establish the separating method of series of cells from human peripheral blood, so that the scientific research and clinical research could be carried out were specifically. Methods 10 cases of normal peripheral blood were collected and T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) were stained with florescence conjugated antibodies. The positive cells were sorted by FACS. Results Before sorting the peripheral blood, the proportion of the T lymphocyte(CD_3~+) and granulocyte (CD_(15)~+) in the leukocyte is 48.8 % and 30.8 %; after sorting by FACS, the purity of T lymphocytes (CD_3~+) is up to 98 % and the recovery is about 95 %; the purity of granulocyte (CD_(15)~+) is up to 97 % and the recovery is about 96 %. Conclusion The FACS could Mlow us to quickly sort T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) with higher recovery and higher purity from the peripheral blood.

Overexpression and clinical implication of MDM2 oncogene in acute leukemia / 白血病·淋巴瘤

Objective To study the over-expression and clinical implications of the oncogene MDM2 in acute leukemia (AL). Methods The expression of MDM2 gene in 100 patients with newly diagnosed and relapse or refractory AL and 20 healthy as control was measured by relative quantitative reverse transcriptase polymerase chain reaction (RT-PCR),then the results was measured by χ2-test,t-test and one-way ANOVA to compare expession positive rate and intensity of MDM2. Results Among 100 patients,fifty-eight had the high expression of MDM2 gene (58 %). The expression level of MDM2 gene in patients was higher than that of health controls(P ＜0.05). The expression positive rate of MDM2 is higher in poor outcome group (67.9 %,19/28)than that in general outcome group (33.9%,19/56) (P＜0.05). Conclusion Our results suggest that the expression of MDM2 gene plays an important role in the pathogenesis and poor outcome of AL.

Clinical significance of cyclooxygenase-2 and BCRP expressions in non-Hodgkin lymphoma / 白血病·淋巴瘤

Objective To investigate the correlation and significance of cyclooxygenase-2 (COX-2)and the breast cancer resistance protein (BCRP) expressions in non-Hodgkin lymphoma(NHL). Methods Ten patients with reactive lymph nodes (RLN) were considered as a control group. Compared with β-actin as internal control, the BCRP mRNA expressions of 61 NHL samples and 10 lymph tissues in control group were detected by semi-quantitative revere transcription polymerase chain reaction (RT-PCR) assay, while the expressions of COX-2 protein of the above specimens were detected by SP immunohistochemistry. Results The positive rates of COX-2 and BCRP in NHL were 50.8 %(31/61) and 45.8 % (28/61), respectively , and were higher than those in the control group (P ＜0.05). The expression of COX-2 was statistically positive correlated to that of BCRP (X2 =8.795, r=0.355, P＜0.05). The expressions of COX-2 and BCRP were not correlated to clinical and pathological factors, such as age, sex, IPI, LDH, β2-microglobulin level and Ann Arbor stage, however, the expression of BCRP was statistically correlated to chemotherapy efficacy.Conclusion BCRP may be involved in multi-drug resistance (MDR) of NHL, so it may contribute to the assessment of chemotherapy and prediction of NHL. Since there is a strong correlation between COX-2 expression and MDR in NHL, the application of COX-2 inhibitors may enhance sensitivity of chemotherapy.

Detection methods and clinical significance of chimerism after hematopoieticstem cell transplation / 国际肿瘤学杂志

Sequential monitoring of chimerism after hematopoietic stem cell transplantation is important for disease prognosis, improving transplantation success rate, and reducing relapses of the original diseases. The application of molecular genetic markers in combination with quantitative PCR to the assessment of chimerism,in particularly the dynamic quantitative analysis of lineage-specific chimerism, can provide important guidance for clinical practice.

Effects of Oxidative Challenge on Defensive Enzyme and Cofactors Level in Alkalibacterium sp.F26 / 微生物学通报

In this paper,a slight halophilic alkaliphile swain,Alkalibacterium sp.F26,which produced high level of intracellular CAT observed in previous research,was selected as a model microbial to explain the responses of this bacterium to oxidative stress.The results indicated that Alkalibacterium sp.F26 had obvious responses to higher concentration (＞1 mmol/L) of H2O2 than that to lower H2O2 (＜1 mmol/L) challenge from the aspects of defensive enzyme synthesis and cofactors level variation.As for catalase production,the activity increased up to 106.54 U/rag protein which was 1.76 fold of the control when cells were challenged by 3 mmol/L H2O2,but its activity only was 1.13 fold when H2P2 was 100 μmol/L.As far as energy state was concerned,ATP production and NAD+ generation were significantly inhibited from 20.55 μmol/L to 17.80 μmol/L and 69.89 μmol/L to 31.77 μmol/L,respectively,leading to the drop of energy charge from 0.77 to 0.68 and the increase of the portion of NADH/NAD+ from 0.08 to 0.41 in the former case.However,these effects were less distinct under lower concentration of H2O2.Except of the condition of 100 μmol/L H2O2,under which the activation of defensive mechanism resulted in an increase of ATE the level of ATP dropped from 22.69 μmol/L of the control to 22.38 μmol/L and 13.70 μmol/L when challenged by 50 μmol/L and 500 μmol/L H2O2.Besides,the concentration of NADH fluctuated and the NAD+ gradually reduced when H2O2 below 1 mmol/L.