Objective To investigate the effects of oxidative stress on homocysteine and related amino acids metabolism in methionine-loading BRL rat hepatocytes.Methods Cultured BRL rat hepatocytes were divided into control and oxidatively stressed group(100 μmol/L H2O2 was added in culture medium for 2 hours),methionine group(50 mmol/L methionine was added in culture medium for 1 hour),and oxidatively stressed + methionine group(100 μmol/L H2O2 was added in culture medium for 2 hours + 50 mmol/L methionine was added in culture medium for 1 hour).At the end of the experiment,culture fluid was collected.Homocysteine,cysteine,and glutathione were measured by high-performance liquid chromatography,and amino acids were assayed by amino acids analyzer.Results Compared with the control group,the contents of homocysteine[(3.76 ± 0.22)vs.(1.54±0.05)μmol/L,P=0.000]and cysteine[(199.80 ±8.75)vs.(99.11 ±2.47)μmol/L,P=0.000]significantly increased in methionine group,and the contents of homocysteine[(3.84 ± 0.34)vs.(1.54 ±0.05)μmol/L,P=0.000]and cysteine[(200.66±8.60)vs.(99.11 ±2.47)μ mol/L,P=0.000]also increased in oxidatively stressed + methionine group.Compared with oxidatively stressed group,the concentrations of homocysteine[(3.76 ± 0.22)vs.(1.67 ± 0.13)μmol/L,P =0.000],cysteine[(199.80 ± 8.75)vs.(82.64±15.88)μmol/L,P=0.000],and glutathione[(1.50 ±0.14)vs.(1.00 ±0.11)μ mol/L,P=0.011)]significantly increased in methionine group,and the concentrations of homocysteine[(3.84 ± 0.34)vs.(1.67±0.13)μmol/L,P=0.000],cysteine[(200.66±8.60)vs.(82.64±15.88)μmol/L,P=0.000]and glutathione[(1.40 ± 0.30)vs.(1.00 ± 0.11)μmol/L,P =0.028]significantly increased in oxidatively stressed + methionine groups.Compared with the control group,the contents of serine[(12.41 ± 1.51)vs.(24.00 ±2.54)mg/L,P =0.000],glutamate[(33.31 ±0.17)vs.(43.10 ±0.52)mg/L,P =0.000]and glycine[(6.23 ± 0.18)vs.(24.66 ± 10.87)mg/L,P =0.000]significantly decreased,while taurine [(7.99 ±0.16)vs.(6.17 ±0.15)mg/L,P =0.000]increased significantly in oxidatively stressed group.Compared with the oxidatively stressed group,the concentrations of serine[(16.98 ± 0.39)vs.(12.41 ± 1.51)mg/L,P=0.006)]and glutamate[(35.44 ±0.82)vs.(33.31 ±0.17)mg/L,P =0.002]in methionine group significantly increased,while taurine[(3.77 ±0.16)vs.(7.99 ±0.16)mg/L,P =0.000]significantly decreased in methionine group.Compared with the methionine group,the contents of serine[(12.59 ± 0.66)vs.(16.98±0.39)mg/L,P=0.008],glutamate[(30.87±0.60)vs.(35.44±0.82)mg/L,P=0.000]significantly decreased while taurine[(4.37 ± 0.12)vs.(3.77 ± 0.16)mg/L,P =0.001]in oxidatively stressed + methionine group significantly increased.Conclusion Oxidative stress can somehow promote homocysteine production in methionine loading BRL rat hepatocytes,but it is not the main effects.

HPLC method is quick,accurate,and sensitive,and can be applied to the determination of riboflavin in hepatic tissue.

Objective To assess the differences in nutrient composition by chemical analysis method and weighing method,A corps of armed police forces daily dietary nutrient intakes were investigated in Tianjin,to understand the status of the actual intakes of dietary nutrient.Methods A dietary survey was performed using the weighing method and with ref-erence to China food composition database in 2002.The intake of each nutrient was calculated through chemical analysis;Nutritional analysis was used in the collection of duplicate food portions during the dietary survey.Results For most of the main dietary nutrients(protein,fat and carbohydrates),minerals (Na,K,Mg,Ca,Zn,Cu and Se)and vitamins (β-car-otene,vitamin E,thiamine,riboflavin,vitamin C,niacin,and vitamin A),the calculated values of intakes were found to exceed those obtained via chemical analysis.Conclusion Differences of the two methods of dietary survey were ultimately due to the analysed values were below the calculated ones.So it is necessary to improve methods of food storage and trans-portation in order to obtain adequate nutrition and to improve the health of troops.

Objective: To investigate the current status of brucella infection in major occupational groups in Bayannur, China, and to analyze the risk factors for brucellosis. Methods: From January to March, 2015, a questionnaire survey and the serological testing were performed to investigate the current status of brucella infection in 649 workers in 13 slaughter and breeding plants in 7 counties in Bayannur, China. The items in the questionnaire survey included general information, related occupational information, living habits, medical history, and awareness of related knowledge. Results: A total of 112 workers (17.26%) had brucella infection. Brucella infection was associated with sex, age, years of exposure to hazardous substances, job, educational background, usage of protective equipment, the presence or absence of a wound at the exposure site, and whether to smoke during the work clearance (P<0.05) . The multivariate analysis showed that the risk factors for brucellosis were meat processing work (OR=1.812) , >5 years of exposure to hazardous substances (OR=1.363) , improper use or nonuse of protective equipment (OR=1.957) , smoke during work (OR=2.027) , and the presence of a woundat the exposure site (OR= 1.231) ; the protective factors for brucellosis were high school and college education and above (OR=0.521) and high rate of awareness of brucella-related knowledge (OR=0.648) . Conclusion The brucella infection rate is high in the major occupational groups in Bayannur, China. The main influencing factors for brucellosis are job, years of exposure to hazardous substances, usage of protective equipment, whether to smoke during work, and the presence or absence of a wound at the exposure site.

OBJECTIVE:To study 3D-QSAR of pyridine heterocyclic ring PI3K inhibitor as anti-renal cancer drug,and to provide reference for the design and R&D of new anti-renal cancer inhibitors. METHODS:The data of structure and active value (pIC50) of 30 pyridine heterocyclic ring PI3K inhibitors were collected. After Sybyl-X 1.1 software used for molecular superimposition, CoMFA and CoMSIA model were established to investigate three dimensional field, electrostatic field, hydrophobic field,hydrogen bond donor site and hydrogen bond acceptor field of PI3K inhibitor molecule. Sybyl-X 1.1 software was used for molecular docking,and the mechanism of PI3K inhibitor molecule and receptor target protein were analyzed. PyMOL V1.5 software was used to design new PI3K inhibitor molecules. The activity of inhibitor molecules was predicted with CoMFA and CoMSIA model. RESULTS:The cross validation coefficients of CoMFA and CoMSIA model were 0.617 and 0.601, fitting validation coefficients were 0.969 and 0.974,and external predictive correlation coefficients were 0.656 and 0.670, respectively. In CoMFA model, contributions of three dimensional field and electrostatic field were 56.2% and 43.8%respectively. In CoMSIA model,contributions of three dimensional field,electrostatic field,hydrophobic field,hydrogen bond donor site and hydrogen bond acceptor field were 41.0%,31.3%,21.1%,2.4%,4.2%. After molecular superimposition,small steric hindrance,strong positive and hydrophilic groups introduced nearby R1 group of common skeleton could help to enhance the activity of molecules. The results of molecular docking showed that PI3K inhibitor molecule formed three hydrogen bonds with the key amino acids ALA805,VAL882 and THR887 of receptor target protein,with the length of 1.84,1.99,1.99 ?. According to above information,6 new molecules were designed,among which predicted pIC50 of 2 molecules with higher activity were 3.211,3.247(CoMFA method)and 3.238,3.222(CoMSIA method). CONCLUSIONS:Established new CoMFA and CoMSIA model have good prediction ability and statistical stability. Contribution of three dimensional field is higher than that of electrostatic field,and the influence of hydrophobic field on molecular activity can not be ignored. Pyridine heterocyclic ring PI3K inhibitors have strong hydrogen bonding role with receptor target protein. 3D-QSAR can provide reference for the design,reconstruction and drug R&D of new PI3K inhibitor molecule.

Objective To investigate the role of endometrial thickness,uterine artery and spiral artery blood flow parameters,endometrial and subendometrial blood flow on pregnancy outcome in vitro fertilization-embryo transfer (IVF-ET) cycle,compare the difference of the endometrial and subendometrial blood flow typing between the superb micro-vascular imaging (SMI) and power doppler ultrasound (PDUS). Methods Forty-one patients undergoing IVF-ET cycles in our hospital were recruited in this study. Transvaginal color Doppler ultrasound was performed to determine and record endometrial thickness,peak systolic velocity(PSV),end diastolic velocity(EDV),pulse index(PI),resistance index(RI), systolic diastolic ratio(S/D) of uterine artery and spiral artery,and degree endometrial and subendometrial blood flow signals were showed under the two blood flow modes of PDUS and SMI respectively on the day of human chorionic gonadotropin(HCG) injection.The degree of the endometrial and subendometrial blood flow was scored using Applehau classification method.According to the pregnancy outcome,patients were divided into pregnancy group and nonpregnancy group,the difference of the above indicators between the two groups were compared,and the degree of endometrial and subendometrial blood flow under two kinds of blood flow mode were observed.Results ①On the day of HCG injection,patients in pregnancy group had lower RI,PI,S/D of spiral artery than those in the nonpregnancy group (P< 0.05). There were no significant difference between the pregnant group and nonpregnant group in endometrial thickness and the blood flow parameters of uterine artery ( P >0.05). ②On the day of HCG injection,there was significant difference between the pregnant group and nonpregnant group in endometrial and subendometrial blood flow degree with SMI mode ( P =0.005).The patients with type Ⅲ in pregnancy group were most (50% ). ③The endometrial and subendometrial blood flow typing (β= 1.085,OR= 1.085,95% CI=1.040 -8.421, P=0.042) of pregnancy group under SMI mode have an independent predictive value.With increased blood flow typing under endometrial and subendometrial blood flow,the higher the possibility of pregnancy in patients.The ROC curve analysis showed that the area under ROC endometrial and subendometrial blood flow typing SMI mode on the HCG day was 0.746( P =0.008).Conclusions SMI is a new and effective type of Doppler ultrasound technique to evaluate the endometrial and subendometrial blood flow in IVF-ET cycle.On the day of HCG injection,the endometrial and subendometrial blood flow typing in SMI mode may predict pregnancy outcomes in IVF-ET cycles,and guide to choose the best embryo transfer time.

Objective:To investigate the correlation between neutrophil/lymphocyte ratio (NLR) combined with low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio (LDL-C/HDL-C) and severity of coronary lesions in patients with acute coronary syndrome (ACS).Methods:Patients who were diagnosed with ACS due to chest pain and received emergency coronary angiography in the First Affiliated Hospital of University of Science and Technology of China and the Affiliated Hospital of Anhui Medical University from January 2017 to June 2020 were enrolled in the final analysis. The data of gender, age, body mass index (BMI), past history, emergency blood routine indicators [neutrophil (NEU), lymphocyte (LYM), monocyte (MON), eosinophil (EOS), basophil (BAS), red blood cell (RBC), mean corpuscular volume (MCV), blood red cell distribution width (RDW), mean platelet volume (MPV), platelet volume distribution width (PDW)], blood lipid index [triglyceride (TG), total cholesterol (TC), HDL-C, LDL-C, very low-density lipoprotein cholesterol (VLDL-C)], and coronary angiography were collected. The results of coronary angiography were evaluated by the Gensini score. According to the Gensini score, the patients were divided into the control group (Gensini score = 0, 55 cases) and the study group (Gensini score > 0, 889 cases), and then the patients in the study group were divided into the low-Gensini-score group (Gensini score < 66, 419 cases) and the high-Gensini-score group (Gensini score ≥ 66, 470 cases). The differences in the general baseline data of the four groups were compared, and the correlation between the statistically significant data and the Gensini score was linearly analyzed, and then the combined diagnostic factors (NLR combined with LDL-C/HDL-C ratio) were obtained by Logistic regression analysis. The receiver operator characteristic curve (ROC curve) was used to evaluate the predictive value of NLR combined with LDL-C/HDL-C ratio in predicting the severity of coronary artery lesions in patients with ACS. Finally, multivariate linear regression analysis was used to establish the predictive model between NLR combined with LDL-C/HDL-C ratio and Gensini score.Results:A total of 944 patients were finally included. The differences in gender, age, BMI, hypertension, diabetes, smoking history, NEU, LYM, MON, EOS, RDW, TC, HDL-C, LDL-C, NLR, LDL-C/HDL-C ratio between the control group and the study group were statistically significant. The differences in BMI, hypertension, diabetes, smoking history, NEU, LYM, MON, EOS, TG, TC, HDL-C, LDL-C, NLR and LDL-C/HDL-C ratio between the low-Gensini-score group and the high-Gensini-score group were statistically significant. Linear regression analysis showed that compared with other indicators, the correlation between NLR, LDL-C/HDL-C ratio and Gensini score was stronger in the study group ( r values were 0.634 and 0.663, respectively, both P < 0.05). Binary Logistic regression analysis of the indicators related to Gensini score showed that NEU, LYM, HDL-C and LDL-C were independent risk factors for coronary stenosis in patients with ACS [odds ratio ( OR) were 0.189, 10.309, 13.993, 0.251, 95% confidence intervals (95% CI) were 0.114-0.313, 4.679-22.714, 3.402-57.559, 0.121-0.519, respectively, all P < 0.05]. ROC curve analysis showed that NLR combined with LDL-C/HDL-C ratio had higher predictive value in predicting the severity of coronary lesions in ACS patients [area under the ROC curve (AUC) was 0.952, 95% CI was 0.93-0.969], when the cutoff value was -3.152, the sensitivity was 98.20%, and the specificity was 81.60%. According to the results of multivariate linear regression analysis, the prediction model between NLR, LDL-C/HDL-C ratio and Gensini score was established, and the formula was Gensini score = -7.772+15.675×LDL-C/HDL-C ratio+8.288×NLR ( R2 = 0.862). Conclusion:There is a significant correlation between emergency NLR combined with LDL-C/HDL-C ratio and Gensini score in patients with ACS at admission, which has a certain predictive value for the severity of coronary artery stenosis in patients with ACS, and can be used as a predictor for evaluating the severity of coronary artery disease.

Objective: To establish a set of rules for autoverification of blood analysis, in order to provide a way to validate autoverification rules for different analytical systems, which can ensure the accuracy of test results as well as shorten turnaround time (TAT) of test reports. Methods: A total of 34 629 EDTA-K2 anticoagulated blood samples were collected from multicenter cooperative units including the First Hospital of Jinlin University during January 2017 to November 2017. These samples included: 3 478 cases in Autoverification Establishment Group, including 288 cases for Delta check rules; 5 362 cases in Autoverification Validation Group, including 2 494 cases for Delta check; 25 789 cases in Clinical Application Trial Group. All these samples were analyzed for blood routine tests using Sysmex XN series automatic blood analyzers.Blood smears, staining and microscopic examination were done for each sample; then the clinical information, instrument parameters, test results and microscopic results were summarized; screening and determination of autoverification conditions including parameters and cutoff values were done using statistical analysis. The autoverification rules were input into Sysmex Laboman software and undergone stage Ⅰ validation using simulated data, and stage Ⅱ validation for post-analytical samples successively. True negative, false negative, true positive, false positive, autoverification pass rate and passing accuracy were calculated. Autoverification rules were applied to autoverification blood routine results and missed detection rates were validated, and also data of autoverification pass rate and TAT were obtained. Results: (1)The selected autoverification conditions and cutoff values included 43 rules involving WBC, RBC, PLT, Delta check and abnormal characteristics. (2)Validation of 3 190 cases in Autoverification Establishment Group showed the false negative rate was 1.94%(62/3 190)(P<0.001), autoverification pass rate was 76.74%, passing accuracy was 97.47%; Validation of 2 868 cases in Autoverification Validation Group, the false negative rate was 3.38%(97/2 868)(P=0.002), autoverification pass rate was 42.26%, passing accuracy was 92.00%; Validation of Delta check on 288 cases in Autoverification Establishment Group and 2 494 cases in Autoverification Validation Group showed the false negative rates were respectively 1.39% and 2.61%(P<0.001). (3)Three hospitals adopted these rules of autoverification for 25 789 blood routine samples, and found that the average TAT of blood routine test reports were shortened by 24min, 32min and 7min respectively, the rate of samples reported within 30min were elevated by 33%, 53% and 7%. The autoverification pass rates were 72%-74%. Conclusions The application of this set of 43 autoverification rules in blood sample analysis can ensure test quality while shortenTAT and improve work efficiency. It is worth pointing out that for the same analytical systems in this research, validation is necessary before application of this set of rules, and periodic validation is required during application to make necessary adjustment; for different analytical systems, as this research provide a way to establish autoverification rules for blood routine tests.Clinical labs may establish their own suitable autoverification rules on the basis of technological parameters. (Chin J Lab Med, 2018, 41: 601-607)

Objective: To detect the high mobility group A2 (HMGA2) expression in renal carcinoma, and to explore the relationship with clinicopathological features and its significance for prognosis. Methods: 50 renal carcinoma specimens, 50 corresponding adjacent normal kidney tissue samples, and 40 benign renal tumor specimens were used in this study. The expressions of HMGA2 mRNA and protein were detected by RT-PCR, Western blot and immunohistochemical assays, and its relationship with clinicopathological features and prognosis in the renal carcinoma patients was analyzed. Results: The RT-PCR results showed that the relative expression levels of HMGA2 mRNA in the renal carcinoma, benign renal tumor tissues, and adjacent normal renal tissues were 0.84±0.23, 0.19± 0.06 and 0.08±0.04, respectively, and the expression in renal carcinoma tissue was significantly higher than those of the other 2 groups (P<0.01). The Western blot results showed that the relative expression levels of HMGA2 protein in the renal carcinoma, benign renal tumor tissues, and adjacent normal renal tissues were 0.91±0.24, 0.12±0.04 and 0.03±0.01, respectively, and the expression in renal carcinoma tissue was significantly higher than those of the other 2 groups (P<0.01). Immunohistochemical results showed that the expression of HMGA2 protein exhibited brown and tan granular, which mainly distributed in the cell nuclei. Among the 50 cases of renal carcinoma, 34 cases exhibited positive expression, with a positive rate of 68.0%. Among the 40 cases of benign tumor tissues, 3 cases had positive expression, with a positive rate of 7.5%, while among the 50 cases of adjacent normal renal tissues, there was only 1 case exhibiting positive expression of HMGA2 protein, with a positive rate of 2.0%. The protein expression of HMGA2 was significantly higher in the renal carcinoma than in the benign tumors and normal renal tissues (P=0.004). There was no statistically significant difference in the association of HMGA2 protein expressions with age, sex, tumor size and histological type (P>0.05), while significant difference did exist in the association with different statuses of TNM staging and lymph node metastasis (P<0.05). The median time to progression (TTP) in 34 HMGA2 protein-positive patients was (22.36±1.48) months and that of 16 HMGA2 protein-negative patients was (34.55±1.87) months (P<0.05). Conclusions HMGA2 plays an important role in the tumorigenesis and development of renal carcinoma, and may be used as an important predictor for estimating the prognosis of renal carcinoma. HMGA2 might become a new diagnostic and prognostic marker for renal carcinoma.