Objective:To obtain monoclonal antibodies(McAb) against Staphylococcal enterotoxin C2(SEC2) and establish method for detecting SEC2.Methods:The secreted SEC2 from staphylococcus aureus was used as antigen to immune BALB/c mice. Monoclonal antibodies against SEC2 were prepared by normal hybridoma technique. By identifying the characters of McAbs, the quantitative detection ELISA test method were established and were preliminarily applied.Results:Four hybridmas producing antibodies against SEC2 were obtained. IgG isotypes of four McAbs were IgG1. Their binding site was different except two McAbs that shared the same binding site.The McAbs were proved to be specific for SEC2.The sandwich ELISA method had good specificity, sensitivity and reproducibility, and it was founded to be able to detect SEC2 at concentration from 0.5 to 20 ng/ml. Its recovery ranged between 97.8% and 101%,and CV value ranged between 2% and 5%.Conclusion:The prepared McAb against SEC2 can be used for SEC2 immunoassay. This work provides a basis for controlling the quality of JINPUSU and researching staphylococcal enterotoxin.

Objective To reconstruct three-dimensional (3D) image of heart and coronary artery on internet and explore the reliability. Methods After the contrast medium was injected into coronary artery of 2 heart specimens, hearts were scanned by UFCT with enhancement volume. One patient's UFCT images of coronary artery angiography were also chosen.Then the primitive images were transposed format, removed noise, specified interesting areas. 3D software on server was applied to reconstruct heart and coronary artery. Results The reconstructed 3D images of the heart and coronary artery were realistically displayed, freely rotated and transected.Main parts and branches of coronary artery were similar to those in the images of the UFCT. Conclusion Heart and coronary artery can be reconstructed on internet,which offers a new approach for reconstructing organs and tissues of human being.;

Objective:The major aim of this study is to analyze the point mutation at the codon 54th of MBL in healthy North Huis,and to measure the plasma levels of MBL, and to analyze the association between the mutation frequency and plasma MBL concentrations.Methods:PCR-RFLP was used to detect MBL point mutation.MBL plasma concentrations were measured using MBL Oliger ELISA kit.Results:Frequency of point mutation at the codon 54th of MBL in healthy Huis was 0.15. The plasma MBL concentration was (3.40?2.55)mg/L. There was negative correlation between MBL concentrations and gene mutation frequency in huis(r=-0.67).Conclusion:The relationship between frequency of mutation at codon 54 of MBL gene and the plasma MBL concentrations in healthy Huis is negative correlation.

Objective:To investigate the expression of ras-related C3 botulinum toxin substrate 3 (RAC3) in glioma tissues and its effect on the migration and invasion of glioma cells.Methods:The expression of RAC3 in 57 glioma patients and their adjacent tissues from the First People’s Hospital of Shangqiu was detected by immunohistochemical assay. According to the experimental requirements, brain glioma cells U87MG were divided into experimental group and control group. The experimental group U87MG cells were transfected with RAC3-siRNA plasmid, and the control group U87MG cells were transfected with MOCK-siRNA plasmid. RAC3 mRNA in each group was detected by fluorescence quantitative PCR. The expressions of RAC3 and MMP2 in each group were detected by Western blot. Transwell was used to detect the migration and invasion ability of cells in each group.Results:The positive rate of RAC3 in glioma patients was 89.47% (51/57 cases) , and the expression rate in paracancer tissues was 14.04% (8/57 cases) . The expression rate of RAC3 in glioma tissues was significantly higher than that in paracancer tissues, with statistical significance ( P<0.01) . After siRNA transfection, mRNA expression of RAC3 in experimental group and control group was 1.23±0.20 and 0.43±0.12, and protein expression of RAC3 was 1.19±0.11 and 0.23±0.08, respectively. The expression of MMP2 protein was 1.19±0.11 and 0.23±0.08, respectively. The expression of MMP2 in experimental group was significantly decreased ( P<0.05) . Transwell assay showed that the number of invasive cells in experimental group and control group U87MG cells was (22±5) and (45±8) , and the number of migratory cells was (34±6) and (90±11) , respectively. In experimental group, U87MG cell migration and invasion ability decreased significantly (both P<0.05) . Conclusion:The high expression of RAC3 in glioma tissues may be related to the malignant degree of development, and affect the migration and invasion ability of glioma cells by regulating the expression of MMP2.

Objective To analyze gene mutations and the etiology of six fetuses with osteogenesis imperfecta detected by prenatal ultrasonography.Methods From March 2016 to May 2017,six gravidas of singleton pregnancy and their fetuses that were diagnosed with osteogenesis imperfecta by prenatal ultrasonography were enrolled in this study.Gravida 1 came to the Center of Prenatal Diagnosis of the First Affiliated Hospital of Zhengzhou University for prenatal diagnosis,while the other five were referred to that center after termination to identify genetic defects with their fetal tissues.Next-generation sequencing technology was carried out for exome sequencing in the genomes of six fetuses.Suspected mutations were confirmed by polymerase chain reaction and Sanger sequencing.Two hundred unrelated healthy individuals were analyzed with Sanger sequencing for validation of novel mutations.Results Fetus 1 carried a heterozygous mutation in collagen,type Ⅰ,alpha-1 (COL1A1) gene,c.724G＞C(p.Gly242Arg),which was found in the mother and brother but not in the father.Fetus 2 carried a known heterozygous mutation in COL1A 1 gene,c.2461G＞A(p.Gly821Ser),which was found in the mother but not in the father.Four heterozygous mutations,c.2282G＞A(p.Gly761Asp),c.1002+5G＞A in COL1A1 gene,c.1774G＞A(p.Gly592Ser) and c.3277G＞T(p.Gly1093Cys) in collagen,type Ⅰ,alpha-2 (COL1A2) gene,were respectively carried by fetuses 3 to 6,but not by their parents.Mutations of c.724G＞C(p.Gly242Arg),c.2282G＞A (p.Gly761Asp) and c.1002+5G＞A in COL1A1 gene and c.3277G＞T (p.Gly1093Cys) in COL1A2 gene were four novel mutations,which were not found in the 200 unrelated healthy individuals.The mother of fetus 1 who was highly suspected with osteogenesis imperfecta selected to continue the pregnancy because the family members had mild symptoms.After delivery,cord blood was collected for genetic test and the result was consistent with that of prenatal genetic diagnosis.Fetus 1 had no fractures during a six-month follow-up after birth.Conclusions Mutations in the COL1A1 and COL1A2 genes may be the etiology of osteogenesis imperfecta in these six fetuses.Results of this study could enrich the data on COL1A1 and COL1A2 mutations relating to osteogenesis imperfecta,and provide a basis for genetic counseling.

Objective To analyze the correlation between function of liver and kidney and blood lipid inde-xes and the prognosis of acute cerebral infarction.Methods 100 patients with acute cerebral infarction were selected.Serum levels of uric acid (UA),serum creatinine (Scr),serum triglyceride (TG),total cholesterol (TC),low density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),and blood calcium (Ca2+).Fasting blood glucose (FPG),fibrinogen (FIB),D two polymer (D-D),serum homocysteine (Hcy),total bilirubin(TBIL),direct bilirubin (DBIL),and indirect bilirubin (IBIL)UA,Scr,TG,TC,LDL-C, HDL-C,Ca2+,FPG,FIB,D-D,Hcy,TBIL,DBIL and IBIL were measured in all the patients the next morning after admission.According to the CSS score,the patients were divided into mild group (0 -15 points,35 ca-ses),medium group (16~30 pointsminutes,34 cases),heavy group 3(31~45 pointsminutes,31 cases).The NIHSS score difference betweenin the scores of the two patients at admission and three months after the onset of the disease was calculated.The patients were divided into the improved group(score difference >0 points, 58 cases),no change group (score difference = 0,36 cases) aAnd worsening group(score difference < 0 point,6 cases).The levels of serum UA,Scr,TG,TC,LDL-C,HDL-C,Ca2+,FPG,FIB,D-D,Hcy,TBIL, DBIL,IBILof different severity and different prognosis of patients were compared and the relevance were ana-lyzed.Results The levels of UA,Hcy,FIB,DD and,LDL-C were the highest in the heavy group and the low-est in the light group.The levels of Ca2+,TBIL,DBIL and IBIL were the lowest in the heavy group,and the highest in the light group.The differences of above indexes between the three groups were statistically signifi-cant (P<0.05).There were positive correlations between the severity of the disease with serum levels of UA, Hcy,FIB,D-D and LDL-C (P<0.05),and a negatively correlations with serum levels of Ca2+,TBIL,DBIL and IBIL (P<0.05).The levels of UA,Hcy,and LDL-C were the highest in the worsening group and the low-est in the improved group,the levels of TBIL,DBIL and IBIL were the lowest in the worsening group and the highest in the improved group.The differences between the three groups were statistically significant (P<0. 05).The prognosis was positively correlated with serum levels of UA,Hcy and LDL-C (P<0.05),and nega-tively correlated with serum levels of TBIL,DBIL and IBIL(P<0.05).Conclusion The serum levels of UA, Hcy,FIB,D-D,LDL-C,Ca2+,TBIL,DBIL and IBIL were significantly correlated with the severity of acute cer-ebral infarction.The serum levels of UA,Hcy,LDL-C,TBIL,DBIL and IBIL were also correlated with pro-longed prognosis.

Primary aldosteronism is a common cause of endocrine hypertension. Aldosterone-producing adenoma and bilateral idiopathic hyperplasia are the two major subtypes of primary aldosteronism. Unilateral adrenal hyperplasia as one of the rare subtypes was seldom reported. Herein, we present in detail a patient with unilateral adrenal hyperplasia difficult to be subtyped. By discussing several key points of the diagnosis and treatment process, some useful experiences and information are provided for the clinicians.

Deficiency, stasis, water and toxin are of great significance in the pathogenesis and pathologic evolution of chronic heart failure (CHF). Based on "deficiency, blood stasis, water and toxin", the pathogenesis and treatment of CHF were discussed in this article. It was found that in the pathogenesis, deficiency--deficiency of heart qi and deficiency of heart yang were the origin of the disease, and blood stasis, water and toxin were the markers of the disease. Among them, blood stasis was the central pathological link, and also an important mechanism that could aggravate the disease and cause a vicious cycle; water-phlegm and water dampness were the basic pathological products; toxin-heat toxin, water toxin, and stasis toxin were the final results of disease progress and product accumulation. In terms of treatment, CHF can be divided into four stages: early, middle, late and end. In the early stage, tonifying qi and regulating heart can be used for the treatment of root cause, and promoting blood circulation and water can be used for the treatment of symptoms; tonifying qi and yin and reinforcing the healthy qi, reducing blood stasis, purging turbid, and eliminating pathogenic factors can be used in the middle stage; reducing blood stasis and removing toxic materials should be used in the late stage, supplemented with warming yang and increasing urine excretion; astringing yang,generating body fluids, tonifying qi and yang should be used in the end stage. At the same time of treating by stages, attention should be paid to adhering to a holistic concept and dialectical treatment; pay attention to timing and flexible medication; adopting a combination of Chinese and Western approaches and integrating them.

Objective This study aims to investigate the impact of cultivation time on dendritic cells(DCs)and their derived exosomes′ expression of immune-related membrane proteins(CD80,MHC-Ⅰ,MHC-Ⅱ)and provides experimental evidence for future research.Methods Mouse bone marrow cells were induced to differentiate into DCs using GM-CSF and IL-4,followed by maturation stimulation withTNF-α.Exosomes were extracted using ultracentrifugation.Western blot and Amnis image flow cytometry were used to identify exosomes derived from mouse DCs.Amnis image flow cytometry was used to detect the expression of immune-related proteins CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ in mouse DCs and their exosomes.Results After 5 days of in vitro cultivation,more than 50%of dendritic cells expressed CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ,reaching the highest level on day 13.The positivity rates were as follows:CD80(97.29±0.63)%,CD11c(92.31±1.18)%,MHC-Ⅰ(97.91±0.49)%,and MHC-Ⅱ(97.91±0.49)%.The differences were statistically significant(P<0.001).The expression gradually decreased after day 13,but approximately 80%of DC cells still expressed MHC-Ⅰ and MHC-Ⅱ immune molecules on day 30.The expression levels of CD80,CD11c,and MHC-Ⅱ on the exosome membrane were highest on day 5 and then decreased overall with prolonged cultivation time,except for MHC-Ⅰ molecules.The differences were statistically significant(P<0.01).Conclusions In vitro-cultured mouse dendritic cells express high levels of immune-related membrane proteins and can be stably maintained for a long time under suitable culture conditions.The secreted exosomes also carry abundant immune-related membrane proteins,but no significant correlation was found between the immune-related proteins on the dendritic cell surface and the exosome membrane surface.

Objective To investigate the prevalence of metabolic syndrome(MS)and its components among people aged over 15 years in Guangxi Zhuang Autonomous Region and to compare the difference between Zhuang and Han populations.Methods Adopting cluster sampling,a survey of diabetes mellitus was conducted in Guangxi from 2003 to 2005.A total of 27 240 subjects aged over 15 years with complete data,including background information of each individual,blood pressure,lipid profile,plasma glucose,blood uric acid and fasting insulin were analyzed in this study.The prevalence of MS and its components were analysed in Han and Zhuang Chinese in Guangxi.The criteria of International Diabetes Federation(IDF)in 2005 and the China Diabetes Society(CDS)in 2004 were applied for diagnosis.Results(1)The crude prevalence rates of MS according to IDF definition were 13.15%in total,12.41%in male and 14.11%in female respectively.The age- standardized prevalence rates of MS(according to the population composition in China in 2000)were 7.66%in total,7.26%in male and 8.81%in female.The crude prevalence rates of MS according to CDS definition were 10.75%in total,13.45%in male and 7.28%in female respectively and the age-standar-dized prevalence rates of MS were 5.9%in total,7.21%in male and 4.31%in female.The prevalence of MS in total,male and female was increasing with age(P