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 explore the risk factors of local relapse and distant metastasis after radical resection of lung adenocarcinoma.Methods A total of 102 patients with lung adenocarcinoma operated in First Affiliated Hospital of Xinjiang Medical University from January 2005 to January 2010 were collected.The correlation between clinicopathological characteristics and prognosis was evaluated by single-factor and multi-factor analyses.The survival curves were plotted using Kaplan-Meier.Single-factor analysis of statistical difference was tested using Log-rank test.Multi-factor analysis of prognostic factors were produced by COX regression proportional hazards model.Results In the whole group,1,2,3 and 5 year disease-free survival rates were 74.30％,58.00％,51.50％ and 44.90％ respectively,and the median disease-free survival was 30 months.Single-factor analysis showed that tumor size (x2 =9.951,P =0.002),clinical type (x2 =8.460,P =0.004),differentiated degree (x2 =4.807,P =0.028),lymph node metastasis (x2 =40.516,P =0.000),pathological stage (x2 =38.769,P =0.000) were prognostic factors for local relapse and distant metastasis in postoperative patients with lung adenocarcinoma.Muti-factor analysis showed that tumor size (OR =1.943,95 ％ CI:1.091-3.463,x2 =5.082,P =0.024),differentiated degree (OR =2.570,95 ％ CI:1.451-4.552,x2 =10.467,P =0.001),lymph node metastasis (OR =3.196,95％ CI:1.037-9.849,x2 =4.096,P =0.043) were independent prognostic factors for local relapse and distant metastasis in postoperative patients with lung adenocarcinoma.Conclusion Tumor size,differentiated degree and lymph node metastasis are independent prognostic factors for local relapse and distant metastasis in postoperative patients with lung adenocarcinoma.

Objective To study a processing method for EEG signals mixed with EOG and ECG signals disturbance.Methods First,the EEG was denoised by the hard threshold method,the soft threshold method,the compromise threshold method and the ? law threshold method in the second generation wavelet,and then the denoised EEG which still contained EOG and ECG was separated by fast independent component analysis( FastICA) algorithm.Results The ? law threshold method of the second generation wavelet had better denoising effect and FastICA algorithm had more ideal separate performance.Conclusion It is an effective preprocessing method for EEG in denoising with the ? law threshold method of the second generation wavelet and then in separating disturbance of independent source with FastICA algorithm.

Objective To compare the clinical effects and adverse effects of microwave ablation (MWA) with sorafenib and sorafenib monotherapy in the treatment of advanced-stage hepatocellular carcinoma (HCC).Methods Medical records and follow-up information of 57 patients with advanced-stage HCC were retrospectively reviewed.25 patients were treated with MWA combined with sorafenib (combined group),and 32 patients were treated with sorafenib monotherapy (monotherapy group).The end points were therapeutic effect,progression-free survival (PFS),overall survival (OS) and adverse reactions.Results The objective response rate in the combined group was similar to the monotherapy group (16.0％ vs.3.1％,x2 =1.521,P =0.217).The disease control rate in the combined group was significantly higher than that in the monotherapy group (80.0％ vs.50.0％,χ2 =5.429,P =0.020).The median PFS in the combined group was longer than that in the monotherapy group (6.0 months vs.3.2 months,x2 =7.675,P =0.006),but the median OS was similar (11.5 months vs.8.5 months,x2 =2.480,P =0.115).The serious adverse reactions were similar between the two treatment groups (44.0％ vs.34.4％,x2 =0.549,P =0.459).Conclusion MWA plus sorafenib is superior to sorafenib alone with respect to PFS in patients with advanced-stage HCC,although it may not improve OS,with no increased risk of serious adverse reactions.

AIM: To study the possibility that yellow wine improves the pathological changes of atherosclerosis in vivo. METHODS: Six weeks old LDL receptor knockout mice (n=48) on a high-fat and L-methionine diet developed plasma hyperhomocysteinemia and atherosclerosis. The animals were randomly divided into yellow wine group, red wine group, ethanol group and control group (n=12 in each group) and were sacrificed after 14 weeks. The levels of plasma lipids and homocysteine in serum were examined. The morphological changes of aorta artery and the atherosclerosis of aorta sinus were observed under microscope. The expression and activation of matrix metalloproteinase-2 (MMP-2) were determined by the method of immunohistochemistry. RESULTS: No significant difference of plasma total cholesterol, triglyceride or high density lipoprotein cholesterol among groups was observed. Plasma homocysteine was significantly decreased in yellow wine group as compared to other three groups (P<0.01). Compared to ethanol and control groups, use of yellow wine and red wine significantly reduced the atherosclerosis lesion area (P<0.01). However, no significant discrepancy between the yellow wine group and red wine group was found. Compared to control group, the expression of MMP-2 in yellow wine group, red wine group and ethanol group decreased by 26.3%, 27.6% (P<0.01) and 5.7% (P>0.05), respectively. The activity of MMP-2 in yellow wine group, red wine group and ethanol group decreased by 31.7%, 32.5% (P<0.01) and 6.7% (P>0.05), respectively. CONCLUSION: Yellow wine and red wine inhibit the expression of MMP-2 and improve the pathologic changes of atherosclerosis, indicating that they have benefic effects on cardiovascular system.

Objective To construct a prokaryotic expression vector of Salmonella paratyphi A ompA gene, and to determine immunogenieity and immonuprotection of the recombinant expressed product rOmpA and carrying frequency of ompA gene in S. paratyphi A isolates. Methods ompA gene was amplified from a clinical S. paratyphi A strain JH01 by PCR, cloned into T-A cloning vector, and then the prokaryotic expression vector was constructed. The rOmpA expression was determined by SDS-PAGE. Antigenicity and immunoreactivity of rOmpA were determined by immunodiffusion test, Micro-Widal's test and Western blot assay, ompA gene exiting in 98 S. paratyphi A isolates was detected by PCR. By using immunoprotective effect of rOmpA against the challenge of S. paratyphi A strain 50001 in mice were determinded. Results Both the nucleotide and putative amino acid sequence identities of the cloned ompA gene were 100% com-pared to the reported corresponding sequences. The expression yield of rOmpA was approximately 65% of the total bacterial proteins, rOmpA was able to induce specific antibody in rabbits, and reacted efficiently with rabbit antisortm or the antiserum against whole cell of S. paratyphi A detected by Western blot. 94.9% (93/98) of the S. paratyphi A isolates had ompA gene. 41.7% (5/12) and 58.3% (7/12) of the mice im-murtized with 100 and 200 μg rOmpA were survival after lethal challenge with S. paratyphi A strain 50001. The titer of antibody to the H antigens of S. paratyphi spp in the sera from rOmpA immunized mice and sur-viral mice in the S. paratyphi A challenge detected by Micro-Widal's test was 1:5-1:40. Conclusion ompA gene has an extensive distribution in clinical isolates of S. paratyphi A. rOmpA possesses an immunogenicity and a certain immunoprotective effect, and can be used as the candidate antigen in genetic engineering vaccine.

Objective: To analyze the diagnosis and therapy for double-hit lymphoma (DHL) and triple-hit lymphoma (THL). Methods:This study involves three patients with follicular lymphoma (FL) that transformed into DHL or THL. These patients were ad-mitted to our hospital between January 2011 and December 2012. All patients were diagnosed by immunohistochemistry and fluores-cence in situ hybridization (FISH). Results:One FL patient transformed into THL and died after 3 months. The other two FL patients who transformed into DHL and who received R-CHOP and R-ESHAP regimens still failed to achieve complete remission. Conclusion:DHL is a rare type of lymphoma that usually involves the bone marrow and central nervous system. This condition is highly resistant to intensive chemotherapy. Part of the DHL cases result from FL. FISH is important for diagnosis. However, a standard treatment for this type of lymphoma remains lacking.

Objective To evaluate the effect of dexmedetomidine on the quality of intraoperative wake-up test in the patients undergoing balloon occlusion test of the internal carotid artery.Methods Forty-two patients of either sex with intracranial aneurysm,aged 57-78 yr,weighing 53-86 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,scheduled for elective balloon occlusion test of the internal carotid artery under general anesthesia,were assigned into 2 groups (n =21 each) using a random number table:propofol conbined with remifentanil group (group PR) and dexmedetomidine combined with propofol and remifentanil group (group DPR).In group DPR,dexmedetomidine was intravenously infused over 15 min in a loading dose of 0.5 μg · kg-1 before induction of anesthesia,followed by an infusion of 0.3 μg · kg-1 · h-1 throughout surgery.Propofol and remifentanil were given by target-controlled infusion (TCI) after infusion of the loading dose.The patients were mechanically ventilated after placement of the laryngeal mask airway.Maintenance of anesthesia was as follows:propofol and remifentanil were given by TCI with the target plasma concentrations of 0.5-1.0 μg/ml and 1-3 ng/ml,respectively,in group DPR;propofol and remifentanil were given by TCI with the target plasma concentrations of 3-5 μg/ml and 3-6 ng/ml,respectively,in group PR.Bispectral index (BIS) value was maintained at 40-60.Before wakeup test,propofol infusion was stopped and the target plasma concentration of remifentanil was decreased to 0.5 ng/ml in two groups,and the infusion rate of dexmedetomidine was decreased to 0.1 pg · kg-1 · h-1 in group DPR.The wake-up time was recorded and the wake-up quality was assessed.After admission to the operating room (T0,baseline),at 10 min before wake-up test (T1),immediately after patients were wakened (T2),at 10 min after patients were wakened (T3) and at the end of wake-up test (T4),the mean blood pressure (MAP),heart rate,respiratory rate (RR),SpO2 and BIS values were recorded.The development of intraoperative awareness,emergence time,postoperative agitation,nausea and vomiting,regurgitation and aspiration and severe pain was recorded.Results MAP,heart rate,SpO2 and RR were all within the normal range during wake-up period in two groups.Compared with the baseline at To,MAP was significantly decreased at Ti,3,4 in group PR,and BIS value was decreased at T1-4 in DPR and PR groups (P＜0.05).Compared with group PR,MAP was significantly increased at T1.3,BIS value was decreased at T24,the wake-up time was shortened,Ramsay sedation score and wake-up quality were increased,the emergence time was shortened,and the incidence of agitation was deceased (P＜0.05),and no significant change was found in verbal rating scale scores assessed after extubation in group DPR (P ＞ 0.05).No cardiovascular events,respiratory depression,intraoperative awareness,postoperative nausea and voniting,regurgitation and aspiration or severe pain was found in two groups.Conclusion Dexmedetomidine can raise the quality of intraoperative wake-up test in the patients undergoing balloon occlusion test of the internal carotid artery.

AIM: To analyze the nucleotide and putative amino acid sequences of PIA genes isolated from N. gonorrhoeae and to construct the prokaryotic expression system of PIA gene.METHODS: The entire PIA genes from 9 strains of N. gonorrhoeae were amplified by using high fidelity PCR. The target amplification fragments were sequenced after T-A cloning. Homology comparison of the nucleotide and putative amino acid sequences of PIA genes from the isolates with the reported sequences in GenBank was then performed. A prokaryotic expression system of PIA gene was constructed. Different dosages of IPTG were applied to induce the expression of the target recombinant protein (rPIA) and 10% SDS-PAGE plus Bio-Rad Agarose Image Analysor was used to determine the expression level of rPIA. rPIA was extracted using Ni-NTA affinity chromatography and the purified effect was detected by SDS-PAGE.RESULTS: In comparison with the reported PIA gene sequences (GenBank No: L19962), the homologies of nucleotide and putative amino acid sequences of PIA genes from the isolates were 99.6%-100% and 99.1%-100%, respectively, which indicated that all the isolates were belonging to serovars IA6. Output of rPIA was as high as 50.1% of the total bacterial proteins. The purified rPIA only showed a single target protein fragment in gel.CONCLUSION: Serovar IA6 is dominant in the local N. gonorrhoeae isolates and sequences of the encoding gene are relatively conserved. The constructed prokaryotic expression system is able to express rPIA with high efficiency, which may lay a foundation for further development of serological detection kit and vaccine of N. gonorrhoeae.

Objective To analyze the characteristics of adverse events following immunization ( AEFI) with varicella attenuated live vaccine ( VarV) in Hebei province during 2012—2014, to evaluate the safety of VarV and to provide supportive evidences for planning chicken pox prevention and control strate-gies in Hebei province.Methods The AEFI data associated with VarV in Hebei province during 2012 to 2014 were collected from the national AEFI information management system and were analyzed by using the method of descriptive epidemiology.Results A total of 239 cases reported during 2012 to 2014 were col-lected from the national AEFI information management system, with a male to female ratio of 1.44 ∶1.The estimated incidence rate was 25.51 per 100 000 doses.One case was serious AEFI with an incidence rate of 0.11 per 100 000 doses.Most of the AEFI cases were children under 3 years old and identified in scattered inhibiting children and kindergarten kids.The main symptoms of common vaccine reactions were fever, local swelling and induration .The rare vaccine reactions were presented as anaphylactic rashes .Conclusion The reported incidence rate of VarV associated AEFI in Hebei province was less than expected.However, the estimated incidence rate of common vaccine reactions in Hebei province was higher than that showed on the Analysis on Surveillance Data of Post-marketing Immunization Safety for Varicella Attenuated Live Vac-cine in China, 2010—2012.Attentions to the AEFI after immunization with VarV should also be paid in fu-ture.