Objective:To lay a foundation of expressing VCTB fusion protein and preparation the oral vaccine for prophylaxis and therapy of H.pylori infection,the prokaryotic expression vectors contained the fusion gene vacA toxic subunit of H.pylori and cholera toxin subunit B(ctxB)was constructed.Methods:The high homogeneity gene fragment was found in comparing VacA toxic subunit gene of domestic typical H.pylori strains.The primers were designed according to vacA and ctxB sequences in the gene bank.VacA toxic subunit gene was amplified by PCR as the template of DNA genome of H.pylori and cloned into plasmid pQE30,which was plasmid pQE30-vacA.The ctxB gene was amplified by PCR as the template of pET32(?)+-ctxB plasmid.The purified ctxB gene was inserted into pQE30-vacA to construct expressing plasmid pQE-vctB of containing vacA and ctxB genes.pQE-vctB was transformed into prokaryotic E.coli Top10.The recombinant plasmid of bacteria cultivated was extracted and purified,and cutted with the incision enzyme to evaluated.Results:The sequence of vacA gene amplified was about 723 bp and the sequence of ctxB gene was about 372 bp.They were consistented with the anticipated length of the DNA.The expressing plasmid pQE-vctB was conformitied with objective gene inserted into pQE30.vctB fusion gene sequenced as 1 092 bp by the sequencing was conformitied with the genebank,and encodes polypeptides of 364 amino acid residues.Conclusion:The prokaryotic expression vectors contained vacA and ctxB fusion gene was constructed successfully and lay a foundation of expression VCTB fusion protein.

DTC in pediatric and adolescent patients has lower incidence and is generally reported to have better prognosis.However,compared with DTC in adults,it is more aggressive and the recurrence rate is higher.Surgery and adjuvant radioiodine therapy can minimize the risk of recurrence.Total or near total thyroidectomy combined with central compartment lymph node dissection is the preferred surgical procedure for most of these patients.Radioiodine therapy for ablation of thyroidal remnant or residual disease is recommended in order to reduce the risk of tumor recurrence,but there is slight chance of increased risk of a sec ond primary malignancy (SPM).Long-term follow-up is therefore recommended for the pediatrics and adolescents with DTC after treatment.Multi-disciplinary collaborative management is needed to optimize treatment efficacy and to minimize adverse effects.

Objective To investigate the correlation between external dose equivalent rate (EDER)and residual radioactivity in DTC patients after 131I therapy.Methods A total of 70 DTC patients (15 males,55 females;average age (45.7±12.8) years) who received 131I therapy from January 2015 to May 2015 were reviewed.Patients were divided into remnant ablation group (Group 1,43 patients) and the group of therapy for persistent disease (Group 2,27 patients).The determination of dose equivalent rate at 1,2 and 3 m from the patients was performed at 1,6,18,24,30,42,48,54,66 and 72 h after the administration of 131 I.Simultaneously,the residual radioactivity was estimated through collecting their urine and monitoring the excretion of radioactivity.The correlation between EDER and residual radioactivity was analyzed.Results The functions of standard EDER (μSv · h-1 · MBq-1) at 1 m from the patients of Group 1 and Group 2 were H=0.044 77e-0.063 8t +0.015 04e-0.396t.and H=0.039 71e-0.0629t +0.014 12e-0.325t,respectively.And the functions of residual radioactivity (MBq) of the two groups were A =A0(0.84e-0.061 7t+ 0.16e-0.158 4t)and A =Ao(0.69e-0.087 7t+ 0.31e-0.047 3t),respectively.There was a positive correlation between EDER and residual radioactivity in DTC patients after the administration of 131I (r=0.95,P＜0.001).The fitting function of the dose equivalent rate at 1 m from the patients was H(μSv/h)=0.048A (MBq).Conclusions The residual radioactivity in DTC patients after 131 I therapy can be estimated by determining the EDER.The dose equivalent rate at 1 m from the patients of lower than 19.2 μSv/h can be used as the limitation of radiation isolation.

Objective To identify clinical features of papillary thyroid microcarcinoma(PTMC) according to patients' age.Methods Seventy-eight patients with PTMC were divided into 2 groups according to age:≥45 years and ＜45 years.The clinical data were retrospectively analyzed.Results The average preoperative thyroglobulin (Tg)level in ＜45 years group was apparently higher than that in ≥45 years group[(138.61 ± 91.87 vs 80.20 ± 85.00) μg/L,P＜0.01].The average tumor size in ＜45 years group was apparently larger than that in ≥45 years group [(0.64 ± 0.24 vs 0.45 ± 0.25) cm,P＜0.01].There were more patients with multiple cancer foci in ＜45 years group than in ≥45 years group (73.53％ vs 45.45％,P＜0.05).And there were more patients with cancer in bilateral lobes in ＜45 years group than that in ≥45 years group(44.12％ vs 18.18％,P＜0.05).There were no significant differences in preoperative thyroid stimulating hormone level,preoperative thyroglobulin antibody (TgAb)level,incidence of capsular invasion of cancer,neck lymph nodes involvement,distant metastasis,and backgrounds of benign thyroid diseases between two groups (all P＞0.05).Conclusion The patients with PTMCs had different clinical features according to age.Hence,clinicians should consider an individualized treatment according to age in order to achieve better therapeutic efficacy.

Objective To analyze the efficacy of 131I therapy for Graves' disease,and to investigate the incidences of complete remission and hypothyroidism after single or multiple treatments.Methods Altogether 2 125 patients with Graves' disease (614 males,1 511 females) aged (39.8 ± 10.2) years received 131I treatment.The diagnostic and therapeutic procedures were carried out as follows:physical examination,determination of thyroid hormones and antibodies,undergoing 131I uptake test to obtain maximum of thyroid uptake value and effective half-life time,ultrasonography,thyroid imaging,calculating 131I therapeutic dosage,131I treatment,follow-up appraisal of curative effect.The data were analyzed by x2 test,optimal scale regression,logistic regression and discriminant analysis.Results The rates of complete remission,hypothyroidism,partial response,and invalid in the 2 125 cases more than half a year after 131I therapy were 54.3 ％,21.3 ％,20.3 ％,and 4.1％ respectively.In all patients the rate of clinical cure (including complete remission and hypothyroidism) and rate of effectiveness were 75.6％ and 95.9％ respectively.The rate of recovery and incidence of hypothyroidism in patients who accepted single 131I treatment were 52.4％ and 21.2％ respectively,while in patients who accepted twice or multiple 131I therapy the respective figures became 66.2％ and 21.8％ accordingly.The influential factors in the effectiveness of 131I treatment included age,thyroid weight,TSH receptor antibody (TRAb),and dose of 131 I per gram of thyroid.Conclusions Patients who did not achieve clinical cure for over 6 months after first 131 I treatment,may receive another131 I therapy to further improve the remission rate.Age,thyroid weight,TRAb etc,contribute to the efficacy of 131I therapy for Graves' disease.The multi-perspective and multi-factor analysis would have the benefit to establish individualized treatment strategy.

Objective To analyze the epidemiological characteristics of pathogens of children with hand , foot and mouth disease (HFMD) in Nantong area, in order to provide the basis for the treatment and prevention of HFMD. Methods Multiplex RT-PCR (fluorescence PCR method) was used to detect EV71, CoxA16 and non-infectious cases of EV71, CoxA16 of children with HFMD in Nantong. VP1 gene of strains isolated from 12 cases of EV71 was amplified and underwent phylogenetic tree analysis. 208 cases clinical data of children with HFMD were analyzed and summarized. Results In 208 cases of children with HFMD swab samples, EV71 of 64 cases were positive (30.8%); CoxA16 of 28 cases were positive (13.5%); 40 cases of other enterovirus were positive (19.2%). Phylogenetic tree analysis showed that EV71 epidemic strains are C4 subtypes in Nantong region. The clinical manifestations of enterovirus infections in EV71 , CoxA16 and other enterovirus are caused by rash, fever and other symptoms, the difference of which was not statistically significant (P > 0.05). In laboratory examination, EV71 infection made white blood cell count higher than CoxA16 and other enterovirus, but the difference was not statistically significant (P>0.05). Conclusions EV71, CoxA16 and other enterovirus intestinal virus are main pathogens of HFMD in Nantong. EV71 C4 subtype is the main popular subtype of EV71 in Nantong. The elevation of white blood cell count of HFMD is the main clinical manifestations and characteristics , but can not serve as the identification of children infected with EV71, CoxA16 and other enterovirus.

Objective To analyze enterovirus 71 (EV71) molecular epidemiology characteristics in Nantong region of Jiangsu , 2014 .Methods The pharyngeal swab samples were selected from 52 children with hand ,foot and mouth disease .After extracting the virus nucleic acid ,the EV71 VP1 gene of the virus were amplified by RT‐PCR amplification .The phylogenetic tree was con‐structed between isolated and EV71 reference strains from other parts of the country .Results 12 strains EV71 VP1 gene were iso‐lated from 52 cases of clinical specimens .The nucleotide and amino acid homology of 12 strains EV71 VP1 gene was 93 .4% -99 .1% ,the phylogenetic tree analysis showed that 12 strains of VP1 gene all belong to C4 genotype C4a subgenotypes .Conclusion The isolated strains in Nantong in 2014 are all the EV71 C4a subgenotypes of C4 genotype ,the same with the most of isolates in re‐cent years .

Objective:To evaluate the changes of metamorphopsia in patients with rhegmatogenous retinal detachment (RRD) who underwent primary vitrectomy and to analyze the correlation between macular microstructure changes and metamorphopsia.Methods:It was a retrospective clinical study. From January 2017 to January 2019, 57 patients (57 eyes) with RRD were examined and diagnosed in the Department of Ophthalmology of Baoding First Central Hospital were enrolled in this study. All patients underwent 23G PPV. All patients had retinal anatomical reduction after the first PPV. The best corrected visual acuity (BCVA), intraocular pressure, slit-lamp microscope, indirect ophthalmoscope, changes of metamorphopsia, and optical coherence tomography (OCT) were examined before PPV and 1, 6, 12 months after the operation. The BCVA examination was performed using the international standard visual acuity chart, which was converted into a logarithm of the minimum angle of resolution (logMAR) BCVA for the record. The macular fovea retinal thickness (CRT) was measured by Carl Zeiss Cirrus HD-OCT5000 instrument from Carl Zeiss Company, Germany, and the microstructural changes in the macular area were recorded, including the presence of the anterior macular membrane (ERM), cystic macular edema (CME), subretinal fluid (SRF), and the integrity of the ellipsoid zone (EZ) and outer membrane (ELM). The value of metamorphopsia (M value) was measured by the M-Chart table. The changes of BCVA, M value, and the microstructure of the macular area before and after operation were analyzed. The correlation between BCVA and M value was analyzed by Pearson correlation. Spearman correlation analysis was used to analyze the correlation between the changes of macular microstructure and BCVA and M values.Results:The mean logMAR BCVA before the operation was 1.15±0.43. At 1, 6, and 12 months after the operation, the mean logMAR BCVA was 0.62±0.17, 0.39±0.18, and 0.34±0.13, respectively. The visual acuity improved significantly after operation compared with before operation, and the difference was statistically significant ( F=119.731, P=0.000). The mean CRT before the operation was 476.0±104.1 μm. At 1, 6, and 12 months after the operation, the average CRT were 299.8±29.9, 272.2±17.8, and 261.0±19.3 μm, respectively. The average CRT after the operation was significantly lower than those before the operation ( F=185.518, P=0.000). At 1, 6, and 12 months after the operation, the mean M values were 0.62±0.54, 0.43±0.41, and 0.32±0.36, respectively; the difference was statistically significant ( F=6.568, P=0.020). After the operation, 48 eyes (84.2%, 48/57) had microstructural abnormalities in the macular area, including EZ and ELM integrity disruption, SRF, CME, and ERM. Correlation analysis showed that M value was significantly positively correlated with BCVA, CRT, and SRF height before operation ( r/ rs=0.672, 0.385, 0.932; P<0.05). There was no correlation between M value and BCVA ( r/ rs=0.503), EZ and ELM integrity ( r/ rs=0.497, 0.472), SRF ( r/ rs=0.416), CME ( r/ rs=0.821) and ERM ( r/ rs=0.632) after operation ( P>0.05). Conclusions:The BCVA is significantly increased and the metamorphopsia is improved after anatomically successful RRD surgery. Postoperative M value is highly correlated with preoperative BCVA, CRT, and SRF.

Objective The purpose of the present study is to compare the pharmacokinetic and biodistribution properties of 99Tcm N-mercaptopyridine-N-oxide (99 Tcm N-MPO) with 99 Tcm-sestamibi (99 Tcm-MIBI) in normal dogs, and to investigate the potential of 99TcmN-MPO as a myocardial perfusion agent in canines with acute myocardial infarction. Methods Twelve healthy mongrel dogs were injected intravenously with 99TcmN-MPO (n = 6) or 99Tcm-MIBI (n = 6). Tracer kinetics in body fluids were determined by collecting blood of 1 ml via a femoral vein catheter at 30 s, 1,2,3,4,5, 10, 20, 30, 40, 60and 90 min post-injection (p. i.). The collected blood samples were weighed and counted for radioactivity in a γ-counter. Anterior and posterior planar γ-camera images were collected at 10, 20, 30, 60, 90, and 120 min after injection, with organ uptake quantified by region-of-interest (ROIs) analysis. For comparison, 99Tcm-MIBI was also evaluated in the same twelve dogs. Canine infarct models were set up by micro-invasive interventional embolization. SPECT images in the canine infarct model were collected 24 hours after myocardial infarction at 30 min and 60 min after the administration of 99Tcm N-MPO (n = 5) or 99Tcm-MIBI (n = 5). Results Both of 99Tcm N-MPO and 99Tcm-M1BI had a rapid blood clearance with less than 50% of initial radioactivity remaining at 1 min [99TcmN-MPO: (35. 77 ± 6. 31)% ID/mg ,99Tcm-MIBI (34. 46 ± 6. 83) % ID/mg] and less than 5% at 30 min p. i. [99Tcm N-MPO(3. 11 ± 1.44) % ID/mg,99Tcm-MIBI (2.93 ±0. 39)% ID/mg] . After injection, 99TcmN-MPO showed significant accumulation in the myocardium and prolonged retention. This rapid liver clearance of 99TcmN-MPO led to favorable heart-to-liver ratios, reaching values of 0. 54 ±0. 06 at 10 min, 1.02 ±0. 06 at 30 min, and 1.38 ±0. 06 at 60 min p. i.In contrast, the heart/liver ratio of 99Tcm-MIBI remained low at all time points (0. 46 ± 0. 03 at 10 min,0. 63 ±0. 03 at 30 min, and 0. 62 ± 0. 12 at 60 min p. i.). SPECT imaging studies in canines with acute myocardial infarction indicated that good visualization of the left ventricular wall and perfusion defects could be achieved at 30 min after administration of 99TcmN-MPO, but not 99Tcm-MIBI. Conclusion The combination of high heart uptake and rapid liver clearance makes 99TcmN-MPO a promising new radiotracer for myocardial perfusion imaging.

Objective To investigate the efficacy and influential factors of 131I treatment for lung metastases from DTC.Methods Fifty patients (18 males,32 females;age (40.8±13.2) years) with lung metastases from DTC who underwent 131I treatment from October 2007 to December 2012 were retrospectively analyzed.The efficacy of 131 I treatment was assessed using 131I imaging and determination of serum Tg level after 6 months.The possible factors affecting efficacy included patients' age,gender,operation method,pathological classification,the diagnostic time of pulmonary metastasis,serum Tg level at diagnosis,131I uptake pattern,characteristics of other imaging modalities,cervical lymph node metastases and extrapulmonary distant metastases (assign 1 for metastases,0 for no metastases).Univariate and multivariate analyses (Student t test,Fisher exact test and logistic regression) were performed to investigate the factors.Results The rates of complete remission,partial response and invalid of 131I treatment were 20％ (10/50),74％ (37/50) and 26％ (13/50) respectively.Univariate analysis showed that age(t =2.019,P＜0.05),gender (P =0.032),serum Tg level at diagnosis (t =2.646,P＜ 0.05),findings of other imaging modalities (P =0.039),and extrapulmonary distant metastases(P=0.023) were the factors influencing outcome of 131I treatment.Multivariate logistic regression analysis showed that the influential factors included age,serum Tg levels and extrapulmonary distant metastases.The regression equation was as follows:logit P =2.127-0.056× age-0.163×Tg level-1.280×extrapulmonary distant metastasis (x2=10.484,P＜0.001).Aged patients,a significant increase of Tg level and extrapulmonary distant metastases indicated a poor prognosis.Conclusions 131I treatment is an effective method for lung metastases from DTC.The patients with younger age,lower Tg levels,no other distant metastases had good response to 131I treatment.