To investigated the toxin genes distribution and molecular characteristics of Vibrio parahaemolyticus from pa‐tients in Ningbo ,V .parahaemolyticus strains were collected from patients with food poisoning and diarrhea .Thermostable di‐rect hemolysin gene (tdh) and TDH‐related hemolysin gene (trh) were detected by polymerase chain reaction (PCR) .Molecu‐lar characteristics were acquired by multi‐locus sequence typing (MLST ) .Of 248 clinical strains were isolated from 2006 to 2012 .Forty‐eight strains were selected to detect virulence genes and MLST genotyping .Forty‐two isolates were detected as tdh+ and 11 isolates were detected as trh+ .There were 9 STs and one undifferentiated type in Ningbo clinical strains .Thirty‐two strains were classified into ST3 ,5 strains into ST265 and 3 strains into ST120 .ST265 was found in Ningbo strains com‐pared with strains from other regions of China .Strains with tdh+ accounted for the majority in Ningbo clinical strains .Twen‐ty‐five strains of ST3 clone were tdh+ /trh‐.There were 9 STs coexsited in Ningbo clinical strains .ST3 clone was dominant , followed by ST265 and ST120 .Strains with tdh+ /trh‐were dominant in the ST3 clone .The unique ST262 was found in Ning‐bo clinical strains .

Objective To investigate the changes of cardiac function and myocardial apoptosis after limb ischemia reperfusion in rabbits.Methods Thirty rabbits were randomly divided into 3 groups:sham-operated control group (group SC),ischemia group (groupⅠ,8 hours of bilateral hind limb ischemia) and ischemia reperfusion group (group IR,4 hours of bilateral hind limb ischemia and 4 hours of reperfusion).Immunohistochemical studies and TUNEL were done to evaluate cell apoptosis.The SOD activity and MDA content in myocardial tissue and plasma LDH activity were determined. The left ventricular function were observed by physiology recording instrument.The cardiac histopathologic changes in experimental rabbits were observed.Results In group IR,the apoptosis index and expression of Bax were significantly increased compared with the other two groups.Similarly,ischemia reperfusion reduced SOD activity and enhanced LDH activity and MDA level,and the left ventricular function decreased in group IR.There was a positive correlation among the MDA content and the LDH activity,the expression of bax,the number of apoptosis cells.Under light microscope, myocardial impairment was found in group IR.Conclusion The myocardial injury occured and the left ventrieular function decreased after limb ischemia reperfusion,and they were associated with oxidative damage and apoptosis.

We investigated the third-generation cephalosporins-resistant Shigella and its genotype in Ningbo,China,providing a basis for disease prevention and control.Pathogenic bacteria were analyzed by direct isolation combined with enrichment culture isolation.Antimicrobial susceptibility was determined by K-B disk diffusion method and PCR was used for detecting multidrug resistance genes like CTX-M,OXA,TEM and SHV.BLAST analysis was used to determine the genotype.Results showed that 69 strains of third-generation cephalosporins-resistant Shigella were detected by drug sensitivity screening,accounting for 74.19％ of ESBLs Shigella.Drug resistance gene CTX-M(CTM-M-1 and CTM-M-9),OXA and TEM were detected.The detection rate were 79.71％,79.01％ and 26.09％ respectively.With no CTX-M-2 and SHV,DNA sequence alignment showed CTX-M-1 group were mainly of CTX-M-15 type besides seven other types;CTX-M-9 group were mainly of CTX-M-14 type besides six other types;49 strains of OXA and 18 strains of TEM were sequenced to be type 1 (OXA-1 and TEM-1 type).The 21 Shigella strains carrying more than two drug resistance genes accounts for 30.43 ％.Shigella in Ningbo has high third-generation cephalosporins-resistance rate and many kinds of ESBLs enzymes were detected.The mainstream enzyme type was CTX-M,meanwhile they also carried a variety of drug resistance genes,which could bring difficulties to disease prevention and control.The high carrying rate of OXA-1 type suggests that we should pay more attention.The detection rate of group B was higher than that of group D,including not only the phenotype resistance but also the drug-resistance genes;these findings will be useful in the study of the drug resistance prevalence of Shigella.

Objective To observe the effects of insulin caliper for blood glucose control on glycemic central tendency, fluctuation and incidence of hypoglycemia in patients with sepsis, and evaluate its application value. Methods One hundred sepsis patients with significant hyperglycemia from December 2015 to December 2017 were selected. All patients needed continuous intravenous insulin infusion to maintain blood glucose. The patients were divided into caliper group and conventional group by random digits table method with 50 cases each, patients of 2 groups adopted an insulin dose modification scheme based on insulin caliper for blood glucose control and paper-based insulin dose modification scheme respectively to control blood glucose. Finally, 92 cases completed the study, including 47 cases in caliper group and 45 cases in conventional group. Blood glucose was measured every 2 hours 0 to 12 hours after intravenous insulin and every 4 hours 16 to 72 hours after intravenous insulin. The incidence of hypoglycemia, insulin dose, ICU time, total hospital stay and hospitalization cost were observed. The proportion of hypoglycemia to total blood glucose measurement, proportion of achieving the glucose control target at each time point, glycemic coefficient of variance, glycemic lability index (GLI) and mean amplitude of glycemic excursion (MAGE) were calculated. Results A total of 1 379 blood glucose values were obtained in caliper group, and a total of 1 332 blood glucose values were obtained in conventional group. There were no statistical difference in blood glucose values 0 to 12 hours after intravenous insulin between 2 groups (P＞0.05). The blood glucose values 16 to 72 hours after intravenous insulin in caliper group were significantly lower than those in conventional group, and there were statistical differences (P＜0.01 or ＜0.05). There were no statistical differences in glycemic coefficient of variance, insulin dose, incidence of hypoglycemia and proportion of hypoglycemia to total blood glucose measurement between 2 groups (P＞0.05). The GLI and MAGE in caliper group were significantly lower than those in conventional group: 12.96 (8.73, 19.58) vs. 23.27 (13.07, 44.61) and (0.66 ± 0.22) mmol/L vs. (0.87 ± 0.28) mmol/L, the proportion of achieving the glucose control target at each time point was significantly higher than that in conventional group: 41.99% (579/1 379) vs. 27.18% (362/1 332), and there were statistical differences (P＜0.01). There were no statistical differences in ICU time, total hospital stay, hospitalization cost, nosocomial infection rate and prognosis between 2 groups (P＞0.05). Conclusions For emergent and critical patients with sepsis, insulin caliper for blood glucose control presents favorable application value for achieving glucose control target, reducing glycemic fluctuation, lowering the incidence of hypoglycemia, low cost and good operability.

OBJECTIVETo provide theoretical basis for artificial cross breeding of Angelica dahurica from Sichuan and Hebei Province, the characteristics of stigma receptivity and the viability and life-span of pollen were studied.

METHODThe viability and life-span of pollen were evaluated by TTC (2, 3, 5-triphenyl tetrazlium chloride) test, and the stigma receptivity was estimated by benzidine-H2O2 method.

RESULTThe pollen viability of A. dahurica from Sichuan and Hebei provinces was increased gradually since the bud stage, but those levels had since subsided after the pollen release from craze antheral. There was a little difference in the pollen viability of A. dahurica from Sichuan at different branches. While the order of the pollen viability of A. dahurica from Hebei was main stem < first-order branching < second-order branching. At room temperature, the pollen viability of both decreased during time of anthers dehiscing but also above 50% after 5 days. Compared with 4 degrees C and room temperature, conservation at - 20 degrees C could extend life of the pollen. The stigma had receptivity in 4th day and reached the highest level in the 6th day after blooming.

CONCLUSIONThe optimum artificial pollination times of A. dahurica was 6 days after blooming and choose the pollen in the peak stage of anthers dehiscing.

Objective:To explore the expression of family with sequence similarity 83 member A (FAM83A) in colorectal cancer, and the effect of FAM83A knockdown on the proliferation of colorectal cancer cells and the related mechanism.Methods:The expression of FAM83A in the tissues of 102 patients with colorectal cancer and its adjacent tissues was detected by immunohistochemistry. HCT116 cells were divided into experimental group and control group. The experimental group cells were transfected with FAM83A-siRNA plasmid, and the control group cells were transfected with MOCK-siRNA plasmid. The mRNA content of FAM83A in each group was detected by fluorescence quantitative PCR. The expressions of FAM83A, P13K, p-AKT and p-mTOR in each group were detected by Western blot. CCK8 assay and clonogenesis assay were used to detect cell proliferation.Results:The positive rate of FAM83A in colorectal cancer patients was 88.23% (90 cases /102 cases), and the expression rate of FAM83A in paracancer tissues was 10.78% (11 cases /102 cases). The expression rate of Fam83a in colorectal cancer tissues was significantly higher than that in paracancer tissues, with statistical significance ( P<0.001). After siRNA transfection, the mRNA expression levels of FAM83A in HCT116 cells of the experimental group and control group were 1.23±0.20 and 0.43±0.12, respectively, and the protein expression levels of FAM83A were 1.19±0.11 and 0.23±0.08, respectively. The expression levels of P13K were 1.21±0.17 and 0.28±0.09, the expression levels of p-AKT were 1.35±0.23 and 0.57±0.18, and the expression levels of p-mTOR were 1.48±0.20 and 1.05±0.14. The expression of P13K, p-Akt and p-mTOR was down-regulated (all P<0.05). The absorbance of HCT116 cells in the experimental group and the control group was 1.09±0.22 and 2.21±0.27, respectively. The cloning rate of HCT116 cells in the experimental group and the control group was 21.6%±2.4% and 62.7%±4.1%, respectively. The proliferation ability of HCT116 cells in the experimental group decreased significantly ( P<0.05) . Conclusions:The expression of FAM83A is significantly increased in colorectal cancer tissues, which may be related to the malignant degree of colorectal cancer. FAM83A affects the proliferation of colorectal cancer cells through the P13K/AKT/mTOR signaling pathway.

OBJECTIVE: To explore the molecular pathogenesis for a pedigree affected with hereditary coagulation factor XII (FXII) deficiency. METHODS: Potential variant of the F12 gene was analyzed by PCR and Sanger sequencing. Expression plasmids were constructed by site-directed mutagenesis based on the wild-type and transiently transfected into 293T cells. FXII:C and FXII:Ag of the expression products were determined in the supernatant and cell lysate. Western blotting was used to verify the identify of the protein. RESULTS: Gene sequencing revealed that the proband has carried 46TT genetype and heterozygous p.Glu502Lys variants in exon 13, and a heterozygous p.Gly542Ser variant in exon 14 of the F12 gene. Transfection experiment suggested that the FXII:C and FXII:Ag of p.Glu502Lys variant in the supernatant were 28% and 24%, compared with the wild-type (100%) and FXII:Ag of cell lysates was 39% compared to the wild-type (100%). The FXII:C and FXII:Ag of p. Gly542Ser variant in the supernatant were 32% and 17% and the FXII:Ag of cell lysates was 59%. CONCLUSION The 46TT genetype, p.Glu502Lys and p.Gly542Ser variants of the F12 gene probably underlie the low FXII level in the proband. As shown by in vitro experiment, the p.Glu502Lys and p.Gly542Ser variants can both inhibit the synthesis and secrection of the FXII protein.
Exons ; Factor XII ; genetics ; Factor XII Deficiency ; genetics ; Heterozygote ; Humans ; Pedigree