Objective To generate the spaO-ompA fusion gene of Salmonella paratyphi A and its prokaryotic expression system,and to determine the immunoprotection of the recombinant expression product rSpaO-OmpA.Methods A flexible peptide sequence was used to link spaO and ompA genes and a prokaryotic expression system of spaO-ompA fusion gene was subsequently generated.SDS-PAGE and Bio-Rad Agarose Image Analyzer were applied to examine the expression as well as the yield of the target recombinant protein rSpaO-OmpA.The antigenicity and immunoreactivity of rSpaO-OmpA were determined using immunodiffusion test,Western Blot assay and micro-Widal's test.By a mouse infection model,the immunoprotection of rSpaO-OmpA against the lethal challenge of S.paratyphi A was determined.In the animal protective test,the recombinant expressed SpaO (rSpaO) and OmpA ( rOmpA ) were used as the controls.Results The generated spaO-ompA fusion gene had 100％ nucleotide and amino acid sequence identities compared to the single spaO or ompA gene.The constructed prokaryotic expression system IPTG E.coli BL21DE3pET42a-spaO-ompA expressed the recombinant protein rSpaO-OmpA.rSpaO-OmpA combined with the antiserum against wholecell of S.paratyphi A to present positive hybridization signal and induced specific antibody in the immunized rabbits.Immunization with 100 or 200 μg rSpaO-OmpA contributed 66.7％ (8/12) or 83.3％ (10/12) immunoprotective rates in mice when the animals were attacked with S.paratyphi A.The immunoprotective rates produced by rSpaO-OmpA were significantly higher than that of equal rSpaO or rOmpA( P＜0.05 ).The sera from rSpaO-OmpA immunized mice presented 1∶5-1∶40 agglutination titers to the H antigens of different S.paratyphi species,and 1∶1-1∶16 immunodiffusion titers to rSpaO,rOmpA and rSpaO-OmpA proteins,respectively.Conclusion The artificially fusion antigen,rSpaO-OmpA,has more powerful immunogenicity and immunoprotection that the equal rSpaO or rOmpA.

The study aims to evaluate the bioequivalence of valsartan hydrochlorothiazide tablets, and to investigate the potential cause of bioinequivalence. This was a single-center study with an open, randomized double-way crossover design. Test and reference preparations containing 160 mg of valsartan and 25 mg of hydrochlorothiazide were given to 36 healthy male volunteers. Plasma concentrations of valsartan and hydrochlorothiazide were determined simultaneously by LC-MS/MS. The pharmacokinetic parameters and relative bioavailability were calculated, while the bioequivalence between test and reference preparations were evaluated. The dissolution profiles of test and reference preparations in four different mediums were determined via dissolution test and HPLC. The similarity was investigated according to the similarity factors (f2). The F(o-t) and F(0-infinity) were (139.4 +/- 65.2)% and (137.5 +/- 61.2)% for valsartan of test preparations. It led to get the conclusion that test and reference preparations were not bioequivalent for valsartan. A significant difference was observed between test and reference tablets in the valsartan dissolution test of pH 1.2 hydrochloric acid solution. The key factor of the bioinequivalence might be that dissolution of valsartan in acid medium has marked difference between two preparations.

To study the metabolite excretion of theophylline, a rapid and specific method by liquid chromatography with heated electrospray ionization tandem mass spectrometry (LC-HESI/MS/MS) method for simultaneous determination of theophylline, 1, 3-dimethyluric acid (1,3-DMU), 3-methylxanthine (3-MX) and 1-methyluric acid (1-MU) in human urine was developed using theophylline-d6 and 5-fluorouracil as internal standards. Selected reaction monitoring (SRM) with heated electrospray ionization (HESI) was used in the negative mode for mass spectrometric detection. After diluted with methanol and centrifuged, the analytes and ISs were separated on a XDB-Phenyl (150 mm x 4.6 mm, 5 microm) column with a mixture of water-methanol-formic acid (30 : 70 : 0.15) as mobile phase at a flow rate of 0.6 mL x min(-1). The linear calibration curves for theophylline, 1, 3-DMU, 3-MX and 1-MU were obtained in the concentration range of 1.0-250 microg x mL(-1), separately. The method herein described is effective and convenient, and can be used for determination of theophylline and its three metabolites. The results showed that urinary excretion ratio of theophylline, 1,3-DMU, 3-MX and 1-MU is approximately 1 : 3 : 1 : 2 in Chinese subjects, which is similar to the reported excretion pattern in Caucasian.

Objective To investigate the effect of the language intervention to reduce patients′discomfort during a colonoscopy examination. Methods A total of 72 subjects who accepted a colonoscopy process in the Digestive Endoscopy Department of the Dalian Central Hospital from September 2014 to December 2015 were divided by random number table method into two groups, 38 for the control group and 34 for the observation group. The control group adopted routine nursing care, while the observation group adopted the language intervention besides the routine nursing care, in accordance with the implementation steps of language intervention for patients who accepted the colonoscopy examination, which means the four stages , the four positions and the four methods. The first stage: the enteroscope entered the anus and arrived the rectum , applied the deep breathing with anal movement method . The second stage: from the rectum to the sigmoid colon, used the induce distraction method. The third stage: stimulated the patients′ excitement at the junctional area of descending colon and transverse colon (the spleen area). The fourth stage: at the junctional area of transverse and ascending colon (the liver ares), used the continuous excitation method. In each stage, individualized language intervention and psychological nursing were given to every patient in the observation group. The cooperating effect of the two groups were evaluated. Results The frequency of pain, the pain scores,the frequency of pressing and the average insertion time of the control group respectively were (8.68 ± 0.83) times,(45.08 ± 6.66) points, (5.73±0.75) times,(372.26±31.77) s, while the observation group were (4.53±0.38) times,(17.07± 3.01) times,(2.05±0.38) times,(268.61±14.51) s, less than the control group ,the t value respectively were 3.81,27.17,3.51,4.66, and the difference was significant (P<0.01 or 0.05). Conclusions Using the language intervention nursing method can significantly relieve the patients′ nervous and anxiety during the procedure, ease the pain and discomfort, shorten the insertion time, improve the subjects′ experience during the colonoscopy examination, make the patients more coordinate with the examination and complete it successfully.

Objective To investigate the clinical characteristics of primary thyroid lymphoma(PTL) and its prognostic factors.Methods Clinical and pathological data of 7 cases diagnosed as primary thyroid lymphoma were retrospectively analyzed.Results All of the 7 patients were diagnosed on thyroidectomy and presented with thyroid nodules,of whom 6 cases were middle-aged to elderly women and some had locally oppressive symptoms.Four cases were diagnosed as diffuse large B non-Hodgkin's lymphoma (DLBCL) and 3 were mucosa-associated lymphoid tissue B-cell lymphoma (MALT).Pathological subtype of mucosa-associated lymphoid tissue B-cell lymphomas and younger patients had better prognosis.Conclusion The possibility of PTL must be kept in mind in the differential diagnosis of thyroid nodules in middle-aged to elderly women.Age and pathological subtype are important prognostic factors.

Objective:To explore the application of OTSU-based self-attenuation correction PET (sacPET) reconstruction technology in 18F-florbetapir (AV45) imaging. Methods:From November 2018 to December 2019, 7 confirmed Alzheimer′s disease (AD) patients (4 males, 3 females, age (69.6±4.5)years) and 3 healthy controls (HC; 1 male, 2 females, age (68.0±4.6) years) were recruited prospectively for 18F-AV45 PET imaging in the First Affiliated Hospital of Sun Yat-Sen University. Original data collected by PET acquisition was processed with sacPET reconstruction and then compared with standard PET images by visual analysis and semi-quantitative analysis. Fisher exact test, Kappa test and Pearson correlation analysis were used to analyze data. Results:In HC group and AD group, the radioactive distribution showed by sacPET images and that by standard PET images were similar, and the contrast of gray-white matter in sacPET images was weaker than that in standard PET images. Moreover, the positive uptake area of the cortex in the AD group was smaller than that in standard PET images. Visual analysis showed 19 positive regions in sacPET images and 22 in standard PET images, with no statistical difference of positive rates of the sub-regions in the cortex between the two PET images (all P>0.05), and the overall consistency of 88.00% (44/50; Kappa=0.75 (95% CI: 0.57-0.94), P<0.05). Semi-quantitative analysis showed that the standardized uptake value ratio (SUVR) of frontal lobe and cingulate gyrus measured by sacPET was lower than that measured by standard PET (0.93±0.06 vs 0.96±0.06 and 0.99±0.04 vs 1.01±0.04; t values: 5.30 and 5.10, both P<0.01), while SUVR of parietal lobe, temporal lobe and occipital lobe measured by sacPET was higher than that measured by standard PET (0.78±0.08 vs 0.68±0.07, 0.97±0.07 vs 0.91±0.08 and 0.94±0.11 vs 0.71±0.12; t values: 6.27, 7.36 and 16.90, all P<0.01). The overall SUVR of sacPET images was significantly correlated with the standard PET images ( r=0.75, P<0.001). Conclusion:For 18F-AV45 imaging, sacPET reconstruction technology can obtain reliable and effective PET images without CT data, but its accuracy and precision still need to be improved.

Objective The purposes of this study were to explore the relationship between personal-ity traits and coping strategies among primary glaucoma patients. Methods 100 patients with primary glaucoma participated the investigation adopting Eysenck Personality Questionnaire (EPQ) and the Medical Coping Methods Questionnaire (MCMQ). Data underwent analysis using descriptive and Spearman rank correlation methods. Results L(lie scale) was (57.83±4.13). The primary glaucoma patients usually took different coping style from norm, facing was (16.91±2.21), avoiding was (15.69±2.01), yielding was (11.54±2.03), all was significantly different than those of norm. L was evidendy negatively correlated with facing, positively correlated with avoiding. Conclusions The personality traits of primary glaucoma pa-tients are different from norm people. They usually take avoiding and yielding coping style, while facing coping style is rarely adopted. Patients with exocentric personality and easy to communicate are apt to adopt positive coping style, while in patients who are identical in emotion and lie, they may take avoiding and yielding coping styles.

Objective:To observe the therapeutic effect of electroacupuncture (EA) at local points plus point injection on House-Brackmann facial nerve function classification grade (H-B) and facial disability index (FDI) in pregnant women with Bell palsy (BP). Methods:A total of 40 eligible BP patients during pregnancy were randomized into a treatment group (n=20) and a control group (n=20). The treatment group was intervened by EA at Sizhukong (TE 23), Tongziliao (GB 1), Sibai (ST 2), Quanliao (SI 18), Yifeng (TE 17), Dicang (ST 4), Jiache (ST 6) and point injection at Zusanli (ST 36) and Yanglingquan (GB 34) on the affected side, 30 min every time, 10 sessions as a course, for 2 courses in total while control group by EA with the same points, manipulation and courses as the treatment group. Results:After the treatment, the H-B classification in the treatment group was better than that in the control group, with a significant inter-group difference (P<0.05); after the treatment, the change of FDI score in the treatment group was significantly different from that in the control group (P<0.05). Conclusion:EA at local points plus point injection can produce a more significant efficacy than EA in the treatment of BP during pregnancy.

Objective To review the clinical data of the first 100 consecutive robotic-assisted laparoscopic radical prostatectomy ( RALRP) performed by a single surgeon , and to provide our experience in passing the learning curve of RALRP .Methods The retrospective study enrolled 100 consecutive patients diagnosed with prostate cancer from June 2012 to November 2014, who had undertaken RALRP , performed by Prof.Gao Xu.The patients aged 67 (50 to 87) years, with a mean PSA level of 20.3 (1.3 to 214.0)μg/L, among which 89 had a clinical staging of T1a to T2b, 4 staged T2c, and 7 staged T3a to T4.Of the patients, 16, 37, and 47 patients were categorized as low , intermediate, and high risk level , respectively. The operation time and blood loss volume were analyzed by moving average method .Results RALRP in all 100 cases were carried out successfully , with no conversion to open prostatectomy .The mean operation time was 221 (120 to 480) min, with an average intraoperative blood loss of 207 (50 to 800) ml; 2 cases received blood transfusion.Positive surgical margin rate was 26%.Eighty (89%) of 90 cases had recovery of continence ( equal to or less than 1 pad used daily ) 3 months postoperatively;10 cases had a follow-up time less than 3 months, and 5 had gained recovery of continence .The operation time and blood loss volume were stable at the 85th -90th case.Conclusion RALRP has a short learning curve , with a promising overall treatment outcome in the early phase of learning .

BACKGROUND:Transplantation of stem cells has a beneficial effect on myocardium revascularization and improving cardiac function after myocardial infarction, and HLA-G protein contributes to the formation and maintenance of the immune tolerance.
OBJECTIVE:To investigate the transplantation effects of human umbilical cord mesenchymal stem cells at different gestational age with different HLA-G expression levels on myocardium revascularization after myocardial infarction in rabbits.
METHODS:Thirty healthy New Zealand rabbits were selected and were randomly divided into human smal gestational age umbilical cord-derived mesenchymal stem cells transplantation group, human ful-term umbilical cord-derived mesenchymal stem cells transplantation group and control group. After the rabbits models of acute myocardial infarction had been established, the former two groups were infused different umbilical cord-derived human umbilical cord mesenchymal stem cells labeled with 5-bromo-2-deoxyuridine into the edge and center of myocardial infarct region by multipoint injection. Rabbits in the control group were subjected to an equal volume of serum-free culture medium.
RESULTS AND CONCLUSION:Four weeks after celltransplantation, 5-bromo-2-deoxyuridine-positive cells were found surrounding the infarct site in both transplantation groups. Myocardial fibrosis and myocardial infarct size were significantly lower in both transplantation groups than those of the control group (P<0.05), and there was a significant difference between the two transplantation groups (P<0.01). The positive staining of factor VII indicated that capil ary density was increased significantly in the smal gestational age umbilical cord-derived mesenchymal stem cells transplantation group as compared with the ful-term umbilical cord-derived mesenchymal stem cells transplantation group (P<0.01), and a sstatistical difference was found between two transplantation groups and the control group (P<0.01). Transplantation of human umbilical cord mesenchymal stem cells with high HLA-G expression increases new capil ary vessels and improves myocardium revascularization. Al indicate that human smal gestational age umbilical cord mesenchymal stem cells have the potential to become the better source of cardiomyocytes transplantation.