This study aims to establish a multiplex TaqMan fluorescence quantitative PCR(qPCR)assay for Pasteurella multocida(P.multocida).Specific primers and fluorescent labeling probes were designed based on the sequences of five podoplanar genes of P.multocida hyaC-hyaD,bcbD,dcbF,ecbJ,and fcbD in the NCBI database.We adjusted the annealing temperature by gradient setting,optimized the primer and probe concentrations by matrix method,constructed standard curves,and performed specificity,sensitivity and reproducibility tests,and finally established mul-tiplexed TaqMan qPCR assays for these five genes.The results showed that the established assay had a good linear relationship between the amplification curves.The sensitivity of this method was high,10-100 times higher than that of ordinary PCR;the specificity was strong,and there was no amplification curve in the DNA detection of eight pathogenic bacteria such as Bacillus,Proteus mirabilis,Staphylococcus aureus,and Rhizoctonia rad iodurans.This assay had a good linear rela-tionship,and the coefficients of variation for Ct values of the inter-and intra-group reproducibility tests were all less than 3％,and the detection rate of this assay was 11.25％higher than the con-ventional PCR assay through the detection of 90 clinical samples.The method established in this study is able to detect P.multocida rapidly and sensitively,which is important for its rapid clinical and laboratory diagnosis.

Objective:To explore the feasibility and value of individualized scheme of dose of iodine contrast medium in enhanced abdominal CT imaging based on liver volume (LV) measurement.Methods:The examination data and technical parameters of 64 patients, who underwent multi-phase dynamic enhanced abdominal CT imaging from February to September 2019 at First People's Hospital of Yunnan Province were retrospectively analyzed. The density of liver on CT pre-contrast images and post-contrast images, LV and average iodine content in the liver (AII) of all subjects were measured. The difference of liver density (ΔHU), total iodine dose of contrast agent injected (IM) and total intrahepatic iodine (TII) were calculated. Pearson correlation analysis was performed to evaluate the association between AII and ΔHU, as well as IM and TII. The difference between the injection volume of contrast agent predicted by the formula and the actual injection volume of contrast agent was tested by paired t test. Results:The ΔHU and AII showed highly positive correlation ( r=0.926, P<0.05). The regression equation was Y AII=0.034X ΔHU+0.296. The TII showed significantly positive correlation with the IM ( r=0.759, P<0.05), and the regression equation was Y IM=3.649X TII+16 486.754. With the liver enhancement ΔHU=50 HU as a reference, the individually dose of contrast medium injection could be deducted based on LV, with Y IM=7.225X LV+ 16 486.754. This formula was introduced into all the subjects to calculate the predicted value of contrast agent application, and the calculated injection volume of contrast agent was (64.27± 4.92) ml. Paired t test was conducted to compare the predicted injection volume of contrast agent with the actual injection volume of contrast agent, and the difference was statistically significant ( t= 6.009, P<0.05). Conclusion:The feasibility of calculating individualized scheme of contrast agent based on LV in enhanced abdominal imaging is verified. This method can be used to predict the enhancement level of liver and to reduce the dosage of iodine contrast agent.

Objective To summarize the pathological survey of time-zero renal biopsy (T0-RBx ) . Methods The material qualities and pathological features were analyzed retrospectively for T 0-RBx (n=176) between March 2008 and May 2016 .According to the source of donor kidney ,T0-RBx specimens were divided into living donors (LD) group (n=137) and Deceased donation (DD) group (n=39) .Furthermore , the DD group was divided into cerebral hemorrhage group (n= 10) and brain trauma group (n= 29) according to the causes of death .The inter-group differences of pathological characteristics and the effects of abnormal pathological lesions on allograft function were observed .Results All T0-RBx specimens contained cortical kidney tissue .The average microscopic length of renal tissue was (0 .39 ± 0 .23) cm and the median glomerular number 11 . The abnormal pathological lesions included glomerulosclerosis (GS ,30 .7 ％ ) , segmental glomerulosclerosis (1 .1 ％ ) ,mesangial increase (MI ,19 .3 ％ ) ,tubular atrophy (TA ,35 .2 ％ ) , acute tubular necrosis (ATN ,9 .1 ％ ) ,vacuolar degeneration of tubular epithelium (27 .3 ％ ) ,losses in tubule epithelial brush border (97 .7 ％ ) , protein cast (25 ％ ) , interstitial fibrosis (IF ,34 .1 ％ ) , inflammation (I ,42 .6 ％ ) ,arteriolar hyalinosis (AH) (26 .1 ％ ) and vascular fibrous intimal thickening (CV ,23 .3 ％ ) .Among them ,23 .9 ％ ,1 .1 ％ ,0 .55 ％ and 0 .55 ％ cases were diagnosed as IgA nephropathy ,immune complex associated with glomerular disease and focal segmental glomerulosclerosis diabetic nephropathy respectively .And the reminders were of ischemic injury .The incidence rates of TA ,IF and I were lower in DD group than those in LD group ( P< 0 .05 ) . However , ATN and vacuolar degeneration of tubular epithelium were higher (P<0 .001) .The incidence of GS was significantly higher in cerebral hemorrhage group than that in brain trauma group (P<0 .01) .No statistical difference existed in other lesions or disease constitution among the groups (P>0 .05) .Further analysis showed GS was related with allograft function at 6/12 months post-transplantation in both LD and DD groups (P<0 .05) .IF and AH were also related to short-term renal function of recipients post-transplantation in LD and DD groups (P>0 .05) .Conclusions T0-RBx may detect the abnormal lesions of donor kidney .Some differences exist in types and degree of abnormal lesions among different donor kidneys .LD group has a higher risk for chronic histological injury such as TA and IF while DD group is more susceptible to acute renal tubular interstitial injury .Thus it is valuable for predicting allograft function post-transplantation .Material quality is essential for ensuring the reliability of T 0-RBx .

To prepare AS1411 targeted nano-ultrasonic contrast agent with liquid core, and to evaluate its ability for ultrasonic contrast enhancement and targeting MCF-7 cell in vitro.
 Methods: The modified solvent evaporation, self-synthesized membrane material and perfluorobrominane (PFOB) was used to form nano-ultrasonic contrast agent with PFOB core (nanoparticles, NP); then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS) catalysis was used to connect AS1411 to the surface of NP to prepare NP-AS1411. The transmission electron microscopy was chosen to check the morphology of NP-AS1411. The size, surface charge, encapsulation efficiency, biocompatibility, the contrast grey value and the stability of NP-AS1411 and NP were compared. Whether AS411 was attached to the surface of NP was checked by gel electrophoresis. Fluorescence microscopy and flow cytometry were performed to examine the targeting ability of AS1411.
 Results: NP-AS1411 was a shell-nuclear structure under the electron microscope. Its size was at (245.4±16.5) nm, which was larger than that of NP (P=0.05). There was no significant difference in surface charge and encapsulation efficiency between NP-AS1411 and NP (P>0.05). In the MTT experiment, the cell viability decreased significantly at high concentration of NP-AS411 (25 mg/mL) after incubation for 24 h compared with the control group (0 mg/mL ) (P<0.05). The contrast gray value of NP-AS1411 was at 86.1+ 6.7, which was significantly higher than that of deionized water (P<0.05), and equivalent to that of NP (P>0.05). The contrast grey value of AS1411-NP was 80.1±9.2 after keeping at room temperature for 24 h, which showed no obviously change comparing with that before the treatment (P>0.05). The size of NP-AS1411didn't change too (P>0.05). The results of gel electrophoresis demonstrated that the AS1411 connecting to the surface of NP was the most when the molar ratio of NP:AS1411 was at 40:1. Flow cytometry analysis confirmed that NP and NP-AS1411 were combined with MCF-7 cells separately but the fluorescence produced by the combination of NP-AS1411 and MCF-7 was more intense.
 Conclusion: The modified solvent evaporation and EDC/NHS catalysis could successfully prepare ultrasound contrast agents with aptamer-conjugated nanoparticles with liquid core. The targeted ultrasonic contrast agents with liquid core possess good ultrasonic contrast enhancement ability in vitro, stability and specificity as well.
Cell Survival ; Contrast Media ; chemical synthesis ; Fluorocarbons ; Humans ; MCF-7 Cells ; Microscopy, Electron, Scanning Transmission ; Nanoparticles ; chemistry ; ultrastructure ; Oligodeoxyribonucleotides ; chemical synthesis

Objective:To evaluate the feasibility and therapeutic efficacy of transhepatic arterial embolization with superparamagnetic iron oxide (SPIO) and lipiodol (LIP) for the treatment of VX2 tumor in rabbits.Methods:Twenty-four rabbits with hepatic VX2 tumors by surgical implantation were randomly divided into 4 groups and treated with transhepatic arterial embolization of 4 different agents as follows (n=6 each):doxorubicin (DOX) group,DOX-LIP group,SPIO-DOX group,and SPIO-DOX-LIP group.Liver function (AST and ALT) was measured at 0,1,3,5 and 7 d after transhepatic arterial embolization.The serum DOX level was measured at 0,5,15,30,60,and 120 minutes after transhepatic arterial embolization.MRI was performed at 7 d after the treatment to assess the distribution of SPIO in the SPIO-DOX group and SPIO-DOX-LIP group,while CT was performed to assess the distribution of LIP in the DOX-LIP group and SPIO-DOX-LIP group.All the rabbits were sacrificed and their livers were removed at 7 d after treatment for the detection of tissue DOX level.The histopathologic examinations were performed including HE staining,Prussian blue staining and TUNEL assay,and then the tumor necrosis percentage and apoptosis index were calculated.Results:Compared to the DOX group,the levels of AST and ALT in other 3 groups were significantly elevated at 1 and 3 d after embolization (P＜0.05).The levels of ALT and AST in the DOX group,DOX-LIP group or SPIO-DOX-LIP group returned to the baseline at day 7,there were no significant differences (P＞0.05).The SPIO-DOX-LIP group exhibited the lowest serum DOX level at all time points up to 120 minutes after embolization (P＜0.05).However,the tissue DOX level in the SPIO-DOX-LIP group was the highest among all groups at day 7 (P＜0.05).The SPIO-DOX group and SPIO-DOX-LIP group showed significantly lower MRI signal intensity of tumors in T2 weighted imaging (T2WI) at day 7.Meanwhile DOX-LIP group and SPIO-DOX-LIP group showed that high-density lipiodol was deposited in the tumors in CT images.Histopathologic findings showed an almost complete central necrosis coagulation of tumors in the SPIO-DOX-LIP group,and the tumor necrosis percentage and tumor apoptosis index were significantly increased in the SPIO-DOX-LIP group compared to those in other 3 groups (P＜0.05).Conclusion:This novel drug-delivery system of SPIO nano-drug carrier together with LIP is safe and feasible when it is used for transhepatic arterial embolization for liver tumor.It provides an excellent MR and CT visualization and improves the therapeutic efficacy for the treatment of rabbit VX2 liver tumor.

Objective To detect the influence of the perfusion quantity and distribution of bone cement by percutaneous ky-phoplasty(PKP) on the early treatment result of thoracolumbar osteoporotic compression fractures(OVCF) .Methods From May 2011 to May 2013 ,62 cases of osteoporotic fractures of thoracic or lumber vertebra were treated by PKP .CT scans were performed postoperatively to analysis the distribution of the bone cement in the vertebra .According to the bone cement distribution on the transverse plane CT film ,the results were classified into four degrees :excellence ,good ,fair and poor .The cases were followed-up regularly .Preoperative and postoperative visual analogue scale(VAS) ,oswestry dysfunction index(ODI) ,height of the operated ver-tebra ,cobb angle ,the incidences of complications during and after the surgery were compared between groups of different degrees of bone cement distribution and different amount of bone cement injection .Results Among the 62 cases ,the follow-up time ranged from 3 to 36 months[average(10 .5 ± 5 .3)months] .In all of the cases ,there was statistically significant difference between the pre-operative and postoperative VAS scoring(P< 0 .05) .3 months after suergery ,there were no statistically significant influence on the results of VAS scoring ,the ODI scoring ,the height lost of the operated vertebra and the improvement of the Cobb angle(P> 0 .05) . In cases of bone cement injection more than 5 mL ,adjacent vertebra fractures happened in 3 cases 6 months postoperatively and 6 cases 12 months postoperatively .In cases of bone cement injection less than 4 mL ,there were only 2 cases of adjacent vertebra frac-tures happened 12 months posoperatively .The degree of vertebra height lost between the bone cement excellent group and poor group was statistically significant in 6 months and 12 months postoperatively .In cases when the distribution of bone cement was ex-cellent ,the improvement of pain and function was significantly different(P< 0 .05) .Conclusion OVCF is treated by PKP .Through conventional operation ,the ultra-early(within 3 months)efficacy is excellent ,in cases of different amount of bone cement injection and different degree of bone cement distribution .However ,with appropriate amount of bone cement ,the more eventfully and sym-metrically the distribution of the bone cement is ,the better of the early clinical results ,probably .

Objective To study the epidemiological features of Cryptococcus neoformans and Cryptococcus gattii isolated from clinical samples in Shenzhen and to elucidate the distribution of species,varieties,genotypes and mating types within the strains tested.Methods The strains involved in this study were 55 cryptococcal strains isolated from our clinical samples.The canavanine-glycine bromthymolblue (CGB) culture was performed to distinguish Cryptococcus neoformans from Cryptococcus gattii.The genotype was characterized by polymerase chain reaction (PCR) fingerprinting with primer M13.The Cryptococcus gattii species and varieties of grubii and neoformans together with two opposite mating type α and a were identified by PCR with variety-specific and mating type-specific primers.The GEF1-restriction fragment length polymorphism analysis was conducted to simultaneously determine the genotype and mating types of strains tested.The sequence type of IGS1 region was analyzed for the VG Ⅱ genotype.Results Of the 55 tested cryptococcal strains,52 were Cryptococcus neoformans,all of which were var.grubii,genotype VN Ⅰ and mating type α.The remaining 3 strains were Cryptococcus gattii,among which,one was genotype VG Ⅰ and mating type α,and two were genotype VG Ⅱ and mating type α.The two VGⅡ genotype strains belonged to the sequence type Ⅱ.Conclusions The strains belonging to the Cryptococcus neoformans var.grubii,genotype VN Ⅰ and mating type α predominate in causative pathogens of cryptococcosis in Shenzhen.Cryptococcus gattii accounts for minority of the cryptococcal isolates,and the highly pathogenic VG Ⅱ genotypes in foreign countries are also characterized.The sequence types of IGS1 region of the two VG Ⅱ strains are in accord with VG Ⅱb sub-genotype.

BACKGROUNDThe prevalence of metabolic syndrome (MetS) increased recently and there was still not a screening index to predict MetS. The aim of this study was to estimate whether brachial-ankle pulse wave velocity (baPWV), a novel marker for systemic arterial stiffness, could predict MetS in Chinese community population.

METHODSA total of 2 191 participants were recruited and underwent medical examination including 1 455 men and 756 women from June 2011 to January 2012. MetS was diagnosed according to the criteria of the International Diabetes Federation (IDF). Multiple Logistic regressions were conducted to explore the risk factors of MetS. Receiver operating characteristic (ROC) curve was performed to estimate the ideal diagnostic cutoff point of baPWV to predict MetS.

RESULTSThe mean age was (45.35±8.27) years old. In multiple Logistic regression analysis, the gender, baPWV and smoking status were risk factors to MetS after adjusting age, gender, baPWV, walk time and sleeping time. The prevalence of MetS was 17.48% in 30-year age population in Shanghai. There were significant differences (χ(2) = 96.46, P < 0.05) between male and female participants on MetS prevalence. According to the ROC analyses, the ideal cutoff point of baPWV was 1 358.50 cm/s (AUC = 60.20%) to predict MetS among male group and 1 350.00 cm/s (AUC = 70.90%) among female group.

CONCLUSIONBaPWV may be considered as a screening marker to predict MetS in community Chinese population and the diagnostic value of 1 350.00 cm/s was more significant for the female group.

Adult ; Ankle Brachial Index ; methods ; Female ; Humans ; Logistic Models ; Male ; Metabolic Syndrome ; diagnosis ; Middle Aged ; ROC Curve

OBJECTIVE:To explore the inhibitory effects of octritide alone or in combination with 5-fluorouracil on the proliferation of human colonic cancer cell line SW480.METHODS:MTT assay was used to detect the absorbance after treatment with octreotide at different concentrations(10-7～10-10 mol?L-1) alone or in combination with 5-fluorouracil on the proliferation of SW480 cells,then the inhibition ratio on SW480 cells was derived.RESULTS:At a concentration of 10-10 mol?L-1,octreotide exhibited a highest inhibition ratio on colonic cancer cell line SW480,at a concentration lower than 10-12 mol?L-1,octreotide had no inhibitory effect on the growth of colonic cancer cell line SW480.The antiproliferative effect of octritide and 5-fluorouracil on SW480 cells was significantly stronger when used in combination than used alone(P

0.05)respectively,but the weekly injection times of the treatment group were reduced and the cost was lower as against the control group and there was no significant increase of adverse reactions in the treatment group.CONCLUSION:It is effective,safe and reliable to use large dose of erythropoietin in the treatment of renal anemia.