To reconstruct the heat-labile enterotoxin subunit B (LTB) gene of Escherichia coil in order to increase the outputs of the prokaryotic expression on recombinant LTB (rLTB), and to determine its immune adjuvant activity on mucosa,the nucleotide sequence of the whole length of LTB gene was synthesized according to the preferred codons of E. coli, and the prokaryotic expression plasmid pET32a-rLTB and its expression system in E. coli BL21DE3 were reconstructed. The recombinant plasmid was extracted and the inserted sequence of rLTB gene was determined. Meanwhile, the expression quantity of the reconstructed rLTB was identified by SDS-PAGE and BioRad agarose image analyzing system, and compared with that of the un-reconstructed rLTB. The abilities of the reconstructed rLTB and the un-reconstructed rLTB to bind with bovine GM1 were determined by means of GM1-ELISA assay. By using the recombinant urease subunit B as antigen, the effects of the reconstructed and the un-reconstructed rLTB on the improvement of immune protection of BALB/c mice infected with Helicobacter pylori strain SS1 and the induction of S-IgAs in infected mice were assayed. The experimental results showed that the expression quantity of the reconstructed rLTB approached upto 35.4% of the total bacterial proteins after induction with 1 mmol/L IPTG for pET32a-rLTB-E. coliBL21DE3 and to be 12.6 times higher than that of the un-reconstructed rLTB (2.8 %). In addition, both the abilities of the recombinant reconstructed LTB and the un-reconstructed rLTB to bind with bovine GM1 could be demonstrated by GM1-ELISA. The immune protection rate of the recombinant urease subunit B in the infected mice was 66.7%; and it could reach up to 91.7% with a significantincrease of the specific S-IgA level, when it was immunized with the reconstructed or the un-reconstructed rLTB. It is concluded that the reconstructed LTB gene in the present study shows a remarkable increased outputs of expression of this gene with a strong immune adjuvant activity on mucosa.

Objective To observe the influences of Shendan Sanjie capsule on chemotherapy effect and immune index to patients with advanced non-small cell lung cancer.MethodsDividing 76 patients with advanced non-small cell lung cancer from department of oncology from January 2014 to May 2016,each with 38 cases.The control group recei-ved western chemotherapy,observation group received western chemotherapy with Shendan Sanjie capsule.The differences of related indicators were compared.Results①After chem-otherapy,in the control group, CD4+decreased,CD8+ increased and CD4+/CD8+ decreased,and before chemotherapy, the difference was significant(P<0.05).In observation group,there was no significant difference in CD4+, CD8+, CD4+ / CD8+ levels.The above indicators of observation group were better than that of control group.②After chemotherapy,RR(CR+PR)rate of observation group were 57.89%, higher than the control group with 36.84%(P<0.05).③During chemotherapy,toxic side effect ratio of observation group was lower than that of the control group(P<0.05).ConclusionComparing Shen Dan Sanjie capsule can adjust patient immunity and reduce side effects of chemotherapy during chemotherapy.

Objective To investigate the effect of morroniside on cycloxygenase (Cox) in the condition of platelet aggregation inducedby adenosine diphosphate (ADP) in rabbits. Methods The levels of Cox induced by ADP in different groups were detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with the control group, all the morroniside groups significantly inhibited theincrease of Cox induced by ADP (P<0.001), which had concentration dependence, and the inhibition rate of high dose group was 30%. ConclusionMorroniside can decrease the level of Cox and it may be the mechanism of morroniside on inhibiting the platelet aggregation inducedby ADP in rabbits.

Objective To understand the predominant β-lactamase genotypes and their carrying modes ofEscherichia coli isolates in Zhejiang province,and the effects of β-1actam antibiotics on inducing or histidine kinase inhibitor closantel (CLO) on inhibiting the expression of β-1actamase genes.Methods Micro-dilution method and E-test were applied to measure the resistant rate and minimal inhibitory concentration (MIC) in E.coli isolates against β-1actam antibiotics.PCR and sequence analysis of PCR products were conducted to detect the β-lactamase genotypes and their carrying modes.Real-time fluorescent quantitative RT-PCR and β-lactamase confirmation test were performed to determine the influence of 1/4 MIC penicillin and cefotaxime,and CLO on the transcription and expression of β-lactamase genes in the resistant E.coli isolates.Results Among the 462 E.coli strains isolated in Zhejiang,285 (61.7％) were resistant to penicillin,ampicillin,cefoxitin,cefotaxim and ceftazidime.In the 285 resistant isolates,the detection rate of TEM or CTX-M β-1actamase gene (83.2％ or 75.1％) was significantly higher than that of KPC,SHV or OXA β-lactamase gene (1.4％-10.2％) (P＜0.01) and the carrying rate of two or more β-1actamase genes (68.8％) was also significantly higher than that of single β-1actamase gene (31.2％) (P＜0.01),and 61.4％ of the resistant isolates carried TEM + CTX-M genes (P＜0.01).Except KPC gene,1/4 MIC of cefotaxim and penicillin induced a rapid increase of TEM-mRNA,CTX-M-mRNA,SHV-mRNA or OXA-mRNA levels (P＜0.01),but 50-500 μg/ml CLO inhibited these levels (P＜0.01).After pre-treatment with 100 μg/ml CLO,82.8％-85.6％ of the resistant isolates became sensitive to β-lactam antibiotics (P＜0.01),while the detection rate of β-lactamases was also decreased from 95.1％ to 16.1％ (P＜0.01).Conclusion TEM and CTX-M are the predominant β-lactamase genotypes in E.coli isolates in Zhejiang and TEM+CTX-M is the predominant carrying mode of β-lactamase genes.Low concentrations of β-lactam antibiotics can up-regulate the expression levels of β-lactamase genes in E.coli through bacterial two-component signaling systems,but this effect can be inhibited by CLO,a histidine kinase inhibitor.

OBJECTIVE: To compare the image quality of three-dimensional time-of-flight (TOF) magnetic resonance angiography (MRA) with sparse undersampling and iterative reconstruction (sparse TOF) with that of conventional TOF MRA. MATERIALS AND METHODS: This study included 56 patients who had undergone sparse TOF MRA for intracranial artery evaluation on a 3T MR scanner. Conventional TOF MRA scans were also acquired from 29 patients with matched acquisition times and another 27 patients with matched scanning parameters. The image quality was scored using a five-point scale based on the delineation of arterial vessel segments, artifacts, overall vessel visualization, and overall image quality by two radiologists independently, and the data were analyzed using the non-parametric Wilcoxon signed-rank test. Contrast ratios (CRs) of vessels were compared using the paired t test. Interobserver agreement was calculated using the kappa test. RESULTS: Compared with conventional TOF at the same spatial resolution, sparse TOF with an acceleration factor of 3.5 could reduce acquisition time by 40% and showed comparable image quality. In addition, when compared with conventional TOF with the same acquisition time, sparse TOF with an acceleration factor of 5 could also achieve higher spatial resolution, better delineation of vessel segments, fewer artifacts, higher image quality, and a higher CR (p < 0.05). Good-to-excellent interobserver agreement (κ: 0.65–1.00) was obtained between the two radiologists. CONCLUSION: Compared with conventional TOF, sparse TOF can achieve equivalent image quality in a reduced duration. Furthermore, using the same acquisition time, sparse TOF could improve the delineation of vessels and decrease image artifacts.
Acceleration ; Arteries ; Artifacts ; Humans ; Magnetic Resonance Angiography

Objective To investigate the clinical manifestations and genetic features of children with papillorenal syndrome caused by PAX2 gene mutation.Methods Clinical manifestations,imaging changes and sequencing data were collected and analyzed from a family with papillorenal syndrome who were diagnosed in Wuhan Children's Hospital in February 2018."PAX2","papillorenal syndrome" and "renal coloboma syndrome" were used as key words to search in China National Knowledge Infrastructure,Wangfang Data Knowledge Service Platform,PubMed and Human Gene Mutation Database up to April 2018.Results A ten years old girl was admitted due to "edema and urine output decreased for one week".Lab showed BUN 25.30 mmol/L,Scr 766.5 μmol/L,Urine protein 3.6 g/24 h.Imaging examination showed bilateral vesical and ureter reflux combined with left duplex kidney and duplication of ureter.Developmental dysplasia of the left hip was also found.The father of the patient had been diagnosed with chronic kidney disease for 10 years and on hemodialysis for 6 years.Next generation sequencing revealed that both the father and daughter carried a heterozygous nonsense mutation in the exon3 c.219C ＞ G(p.Y73X) of PAX2.No Chinese literature ever was reported about papillorenal syndrome.Ninety-four articles in English were retrieved and 177 patients with papillorenal syndrome were confirmed by gene analysis with a total of 92 PAX2 variants.Ten nonsense mutations had been reported.Developmental dysplasia of the hip (DDH) never be reported before.Conclusion Papillorenal syndrome caused by PAX2 mutation can mainly manifest as abnormal development of both kidney and optic nerve,which may be accompanied by other systemic abnormalities,it is rarely reported in China.DDH may be a new phenotype of papillorenal syndrome.

OBJECTIVE: To develop and validate radiomics models based on non-enhanced magnetic resonance (MR) imaging for differentiating chondrosarcoma from enchondroma. METHODS: We retrospectively evaluated a total of 68 patients (including 27 with chondrosarcoma and 41 with enchondroma), who were randomly divided into training group (=46) and validation group (=22). Radiomics features were extracted from TWI and TWI-FS sequences of the whole tumor by two radiologists independently and selected by Low Variance, Univariate feature selection, and least absolute shrinkage and selection operator (LASSO). Radiomics models were constructed by multivariate logistic regression analysis based on the features from TWI and TWI-FS sequences. The receiver-operating characteristics (ROC) curve and intraclass correlation coefficient (ICC) analyses of the radiomics models and conventional MR imaging were performed to determine their diagnostic accuracy. RESULTS: The ICC value for interreader agreement of the radiomics features ranged from 0.779 to 0.923, which indicated good agreement. Ten and 11 features were selected from the TWI and TWI-FS sequences to construct radiomics models, respectively. The areas under the curve (AUCs) of TWI and TWI-FS models were 0.990 and 0.925 in training group and 0.915 and 0.855 in the validation group, respectively, showing no significant differences between the two sequence-based models (>0.05). In all the cases, the AUCs of the two radiomics models based on TWI and TWI-FS sequences and conventional MR imaging were 0.955, 0.901 and 0.569, respectively, demonstrating a significantly higher diagnostic accuracy of the two sequence-based radiomics models than conventional MR imaging (<0.01). CONCLUSIONS The radiomics models based on TWI and TWI-FS non-enhanced MR imaging can be used for the differentiation of chondrosarcoma from enchondroma.
Chondroma ; Chondrosarcoma ; Humans ; Magnetic Resonance Imaging ; ROC Curve ; Retrospective Studies

Compared to conventional cancer treatment, combination therapy based on well-designed nanoscale platforms may offer an opportunity to eliminate tumors and reduce recurrence and metastasis. In this study, we prepared multifunctional microspheres loading I-labeled hollow copper sulfide nanoparticles and paclitaxel (I-HCuSNPs-MS-PTX) for imaging and therapeutics of W256/B breast tumors in rats. F-fluordeoxyglucose (F-FDG) positron emission tomography/computed tomography (PET/CT) imaging detected that the expansion of the tumor volume was delayed (<0.05) following intra-tumoral (i.t.) injection with I-HCuSNPs-MS-PTX plus near-infrared (NIR) irradiation. The immunohistochemical analysis further confirmed the anti-tumor effect. The single photon emission computed tomography (SPECT)/photoacoustic imaging mediated by I-HCuSNPs-MS-PTX demonstrated that microspheres were mainly distributed in the tumors with a relatively low distribution in other organs. Our results revealed that I-HCuSNPs-MS-PTX offered combined photothermal, chemo- and radio-therapies, eliminating tumors at a relatively low dose, as well as allowing SPECT/CT and photoacoustic imaging monitoring of distribution of the injected agents non-invasively. The copper sulfide-loaded microspheres, I-HCuSNPs-MS-PTX, can serve as a versatile theranostic agent in an orthotopic breast cancer model.