PurposeTo investigate the feasibility of using low concentration contrast agent and low tube voltage in the light and moderate weight's abdominal contrast-enhanced CT scan, in order to find an optimal solution to reduce radiation dose and iodine intake.Materials and Methods Forty patients with light weight whose body mass indexes (BMI) were lower than 20 kg/m2 were randomly divided into group A1 (n=20) and group B1 (n=20). Meanwhile, another 40 patients with moderate weight whose BMI ranged from 20 kg/m2 to 25 kg/m2 were randomly divided into group A2 (n=20) and group B2 (n=20). Low concentration contrast agent and low tube voltage (Visipaque 270 mgI/ml, 100 kV) were used in both group A1 and group A2 in abdominal enhanced CT scan. While both group B1 and group B2 used conventional scan solution (Omnipaque 300 mgI/ml, 120 kV) in abdominal enhanced CT scan. Then the contrast noise ratio (CNR), the image quality score and the effective radiation dose (ED) were compared among the four groups.Results The CNR and image quality score at artery phase and portal phase were neither significantly different between group A1 and group B1, nor between group A2 and group B2 (t=-1.539-0.000,P>0.05). The CNR and image quality score of the liver at artery phase in group B1 were signiifcantly higher than those in group A2 and group B2 (P<0.05).Conclusion The solution of using low concentration contrast agent and low tube voltage in contrast enhanced scan can achieve the same high quality abdominal image with reduced iodine intake and radiation, compared with the application of conventional enhanced scan; BMI has rather great impact on image quality score at arterial phase and little impact on that at portal phase. So it is suggested that the protocol of liver contrast-enhanced CT scan may choose reduction of voltage at portal phase so as to reduce radiation.

Objective To establish a simple and rapid detection technique for Oncomelania infected with Schistosoma japonicum(SJ), with high sensitivity and good specificity .Methods The gene fragment of SJ was amplified by PCR , and cloned into the T-vector to construct positive-reference.An isothermal nucleic acid amplification reaction system for detecting Oncomelania infected with SJ was set up , and its sensitivity was analyzed by detecting positive-reference diluted according to geometric proportion , and its specificity by detecting the genomic DNA of relative samples .Then, a corresponding means of purifying nucleic acid was designed to assemble a reagent detecting Oncomelania infected with SJ . This reagent was validated by detecting Oncomelania samples.Results The 213 bp amplified products were obtained and used to construct recombination T-vector for positive reference .An isothermal nucleic acid amplification reaction system was set up for detecting Oncomelania infected with SJ , and the amplification results could be simply determined by color change, with better sensitivity and specificity .The reagents for detecting Oncomelania infected with SJ were assembled , which could detect samples containing only 1% infected Oncomelania.Conclusion A visible detection method for Oncomelania infected with SJ is successfully established and validated .

OBJECTIVETo explore the influences of anti-CD47 monoclonal antibody (mAb) B6H12 on the differentiation and function of cultured dendritic cells (DCs).

METHODSHuman peripheral monocyte derived DCs were propagated with granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin-4 (IL-4) in the presence or absence of soluble B6H12. Flow cytometry was used to analyze the immunophenotypes of cells, semi-quantitative RT-PCR and ELISA methods to analyse the mRNA and protein expression levels of interleukin-12 (IL-12). The antigen-presenting function of DCs was determined in one-way mixed leukocyte reaction by Brdu-ELISA technique.

RESULTSThere was a high expression rate (94% approximately 98%) of CD47 molecules in DCs. The cell immunophenotypes in B6H12 mAb treated and untreated DC groups were as follows: CD80(+) (68.14 +/- 7.41)% vs (89.17 +/- 8.59)%; CD86(+) (67.33 +/- 4.71)% vs (87.27 +/- 3.56)%; CD83 (40.08 +/- 14.80)% vs (72.77 +/- 8.68)%; CD1a(+) (66.45 +/- 4.06)% vs (95.93 +/- 3.03)%; and HLA-DR (40.67 +/- 13.48) vs (98.97 +/- 1.01)%, respectively. The expression levels of mRNA and protein of IL-12 were strongly inhibited in B6H12 mAb treated DC (P < 0.01). The quantity of Brdu was also lower in B6H12 mAb treated DC than that in untreated DCs (P < 0.01).

CONCLUSIONThe anti-CD47 McAb exerts a negative effect on the maturation and functions of cultured DCs.

Adult ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; analysis ; Antigens, CD1 ; analysis ; B7-2 Antigen ; analysis ; CD47 Antigen ; analysis ; immunology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Immunoglobulins ; analysis ; Interleukin-12 ; genetics ; metabolism ; Interleukin-4 ; pharmacology ; Lipopolysaccharides ; pharmacology ; Membrane Glycoproteins ; analysis ; Monocytes ; cytology ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

DNA Methylation ; Glioma ; metabolism ; Humans ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; genetics

Abstract: Objective　To analyze the emergency response and long-term intervention effects after the detection of infectious snails epidemic by loop-mediated isothermal amplification (LAMP) assays in Hannan District, Wuhan City, and to explore the application of LAMP in early surveillance and early-warning of schistosomiasis transmission. Methods Snails picked up by the risk monitoring system in Hannan District were examined by anatomical microscopy and LAMP technology to identify the schistosomiasis infection. Emergency response and intensive intervention were initiated in the environment where positive snails appeared, and the long-term effects were evaluated. Results In May 2018, the infectious snails were detected by LAMP technology in Hannan District, and the positive snails were located in Zhujiacha, Dongzhuang Village, Obstacles and weeds were removed and buried by machine in Zhujiacha. 12 700 m2 of snails were killed by drugs, and the mortality rate of snails was more than 80%; no new seropositive persons were found in the emergency examination within 500 m of the positive snail sites. 506 people were examined in Dong Zhuang Village at the end of the year, and 30 positive IHA cases were detected with a blood positive rate of 5.93%, no positive fecal test was found, and all positive blood test patients took preventive medication. The monitoring results of sentinel rats and wild feces were all negative. Health education was carried out, 7 warning signs were deployed and refreshed, and 500 publicity brochures were distributed. After nearly three years of intensified intervention and monitoring in the villages where the positive environment is located, and the density of snails on the stubborn snail has dropped from 0.094/frame to 0.027/frame, and the positive rate of blood test in Dongzhuang Village has steadily dropped from 5.93% to 3.74%. Conclusions The infected snails missed by microscopy were detected by LAMP in Hannan District, which created conditions for the rapid emergency treatment of environment and elimination of positive snail and improved the sensitivity of the surveillance and early warning system in transmission-interrupted areas.

Classical swine fever virus (CSFV), an enveloped positive-stranded RNA virus in the genus Pestivirus of the Flaviviridae family, is the causative agent of a highly contagious swine disease characterized by symptoms of hemorrhagic fever and immune depression, usually leading to substantial economic losses. The serological methods for detection of CSFV antibody such as ELISA are important means for the diagnosis of CSFV and immune surveillance. It is difficult to obtain CSFV antigen with high quality using traditional method because its titration titer is low in cell culture. CSFV has four structural protein named C, E0, El and E2. The E2 protein contains major antigenic determinants that are conserved between different CSFV strains and involved in neutralization by antibodies. So recombinant E2 protein can be developed as an alternative to the intact viral antigen. So far, CSFV E2 have not been expressed in E. coli with high level. Many factors, such as the secondary structure, the stability of 5' and 3' terminus of gene, the location of SD sequence and the bias of codes, are involved in the expressing level of foreign gene in E. coli . In this study, two sites of the E2 gene sequence were confirmed to be detrimental to its expression efficiency in E. coli through the computer-aided analysis. So they were mutated using recombinant PCR without changing the amino acids sequence of CSFV E2 gene. A plasmid was constructed by inserting the mutated E2 gene into the prokaryotic expression vector pET-28a(+) and named pETE2. The E. coli competent host BL21 (DE3)lysS transformed with pETE2 could express the E2 gene at high level, amounting to 28% of the total protein of the induced recombinant bacteria at the presence of IPTG. Except the hydrophobic transmembrane domain at C terminus, the recombinant E2 protein includes the total aa sequence. So it contains all the potential linear antigen epitopes of E2 protein because hydrophobic aa region can not form epitope. The recombinant E2 protein was CSFV-specific as proved by Western blotting and indirect ELISA. The rabbits immunized with the recombinant E2 can be protect from the challenge of hog cholera lapinized virus. This is the first report that E2 gene is expressed with high level expression in E. coli. In conclusion, it is an effective measure that mutate the CSFV E2 gene to increase its expression level in E. coli. The recombinant CSFV E2 protein possess fine immunonicity and can be used the antigen for the detection of CSFV antibody.
Antigens, Viral ; genetics ; immunology ; metabolism ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Mutagenesis, Site-Directed ; methods ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Viral Envelope Proteins ; genetics ; immunology ; metabolism

AIM To investigate the effect and mechanism of Qigui Yishen Decoction (QGYS,Astragali Radix,Angelicae sinensis Radix,Chuanxiong Rhizoma,Achyranthis bidentatae Radix) on regulating the expression of miR-141 in unilateral ureteral obstruction (UUO) mice with renal fibrosis.METHODS Thirty Balb/c male mice randomly divided into sham-operated group (n =6),UUO group (n =6),Lotensin (50 g/kg) group (n =6),QGYS high dose (50 g/kg) group (n =6),and QGYS low dose (10 g/kg) group (n =6) were conducted UUO surgery to promote kidney fibrosis except the six mice in the sham operation group.After a successive 10-day medication of QGYS and Lotensin to mice by oral gavage on daily basis,all mice were killed to procure renal tissue to observe its morphology and pathology changes by HE staining.The expressions of TGF-β1,ColⅣ,and MMP-9 were analyzed by immunohistochemical method,and the expressions of miR141,TGF-β1 were measured by real-time PCR.RESULTS The obviously pathological injuries including renal interstitial fibrosis were identified by HE staining among the groups intervened with UUO,but the variance in the extent due to different administrations of QGYS and Lotensin was noticed as well (P ＜ 0.05).As compared to the UUO group,high and low dose QGYS groups and Lotensin group achieved an up-regulated expression of TGF-β1 and ColⅣ,and a down-regulated expression of MMP-9 by immunohistochemistry (P ＜ 0.05),and significantly increased Mrna expression of miR-141,and decreased Mrna expression of TGF-β1 by real-time PCR (P ＜ 0.05).CONCLUSION In UUO mouse models,QGYS gives influence to TGF-β1and MMP-9 through inducing miR-141 expression change to decrease abnormal accumulation of ECM,and thus inhibits the progression of renal fibrosis.

China ; Encephalomyelitis, Acute Disseminated ; complications ; therapy ; virology ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; isolation & purification ; Influenza, Human ; complications ; therapy ; virology ; Middle Aged ; Pneumonia, Viral ; complications ; therapy ; virology ; Treatment Outcome

Escherichia coli strain DC1515, deficient in glucose phosphotransferase (ptsG), lactate dehydrogenase (ldhA) and pyruvate:formate lyase (pflA), is a promising candidate for the fermentative production of succinate. To further improve the succinate producing capability of DC1515, we constructed plasmid pTrchisA-pyc with heterogenous pyruvate carboxylase (pyc) from Bacillus subtilis 168 under the Trc promoter and introduced it into DC1515. We used lactose as a substitute of IPTG to induce pyc. We optimized the culture conditions such as the lactose addition time, the lactose concentration and the culture temperature after induction for succinate production. We also explored the effect of lactose supplement during the fermentation. The results showed that pyc can be expressed under lactose induction in the fermentative medium with 15 g/L glucose due to the deficient of ptsG in DC1515. Under optimized conditions, the final succinate concentration reached to 15.17 g/L, which was 1.78-fold higher than that of control strain. If complementing lactose twice to the concentration of 1 g/L during the fermentation, the final succinate concentration could further reach to 17.54 g/L. This work might provide valuable information for gene expression in E. coli strains using lactose as inducer for succinate production in a glucose-medium. Due to the reduced cost, E. coli is becoming a more promising strain for succinate production through fermentation.
Bacillus subtilis ; enzymology ; Culture Media ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Lactose ; pharmacology ; Promoter Regions, Genetic ; Pyruvate Carboxylase ; biosynthesis ; genetics ; Succinic Acid ; metabolism

Animals ; Fatty Liver, Alcoholic ; drug therapy ; Female ; Male ; Phytotherapy ; Rats ; Rats, Wistar ; Saponins ; therapeutic use ; Trigonella