Objective To construct a recombinant live attenuated Salmonella typhimurium nucleic acid vaccine carrying Helicobacter pylori ( H. pylori ) hpaA gene and to detect its immunogenicity. Methods hpaA gene fragment was amplified by polymerase chain reaction (PCR) from the genomic DNA of the standard H. pylori strain and cloned into pUCmT vector. DNA sequence of the amplified hpaA gene was assayed,then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions. The recombinant plasmid was used to transform Escherichia coli DH5?,and the positive clones were screened with PCR reaction and restriction enzyme digestion. Then,the recombinant pIRES-hpaA was transformed to LB5000,then the recombinant plasmid was transformed to SL7207,and the recombinant strain was passaged repeatedly. Finally,recombinant pIRES-hpaA was transfected to COS-7 cells with Lipofectamine TM 2000. The immunogenicity of expressed hpaA protein was determined with SDS-PAGE Western blot. Results The hpaA gene fragment of the 750 base pair was amplified from the genomic DNA,and it was consistent with the sequence of the H. pylori hpaA by sequence analysis. It was confirmed by PCR and restriction enzyme digestion that Helicobacter pylori hpaA gene was inserted into the eukaryotic expression vector pIRES and a stable recombinant live attenuated Salmonella typhimurium nucleic acid vaccine carrying Helicobacter pylori hpaA gene was successfully constructed,and specific strip of hpaA expressed by pIRES-hpaA could be detected with Western blotting. Conclusion The recombinant attenuated Salmonella typhimurium nucleic acid vaccine strain expressing HpaA protein with immunogenicity was constructed,it might be helpful for further investigation concerning the immune action of the nucleic acid vaccine in vivo .

0. 05 ) . Conclusion The cagG gene of Hp was quite conservative, most of the Hp strains in Chinese patients were cagG positive, it has no specific predisposition to different disorders and shows no significant correlation to the extent of gastric mucosal inflammation. Therefore cagG can' t be served solely as a related pathogenic gene for certain diseases.

Objective To construct nucleic acid vaccine encoding Helicobacter pylori (H. pylori) hpaA gene. Methods The genomic DNA of the standard H. pylori strain 17874 was isolated. hpaA gene fragment was amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. The sequence of the amplified hpaA gene was assayed, and then cloned into the eukaryotic expression vector pIRES through a series of enzyme digestion and ligation reactions. The recombinant plasmid was transformed into competent Escherichia coli cells DH5?, and the positive clones were screened by PCR reaction and restriction enzyme digestion . Recombinant pIRES hpaA was transfected into COS 7 cells using Lipofectamine TM 2000, and the immunogenicity of expressed HpaA protein was detected with SDS PAGE and Western blot. Results The 750 base pair hpaA gene fragment was amplified from the genomic DNA and was consistant with the sequence of the H. pylori hpaA by sequence analysis. PCR and restriction enzyme digestion results revealed that the H. pylori hpaA gene was inserted into the eukaryotic expression vector pIRES, suggesting the nucleic acid vaccine pIRES hpaA was successfully constructed. The specific protein strip of HpaA expressed by pIRES hpaA was detected with Western blot. Conclusion The nucleic acid vaccine pIRES hpaA was successfully constructed which may help the further investigation towards the immune action of the nucleic acid vaccine.

Objective To investigate the clinical effects and indications of phacoemulsification and soft intraocular lens(IOL)implantation combined with viscogoniosynechialysis in the management of primary angle closure glaucoma(PACG).Methods Forty-six eyes of primary angle closure glaucoma with co-existing cataract were treated with phacoemulsification and IOL implantation combined with goniosynechialysis.The mean follow-up duration was 9 to 24 months.The main outcome measures were pre-operative and post-operative changes of intraocular pressure,visual acuity,depth of anterior chamber,anterior chamber angle,outflow facility of aqueous humor(C values)and PO/C ratio,and the number of glaucoma eye drops required.Results Compared with pre-operation,intraocular pressure was decreased(paired t test,P

AIM: To explore the nursing cooperation highlights of eight osteopetrosis patients underwent optic nerve decompression via transsphenoidal microsurgical approach instead of routine pathway, and to improve the quality of nursing cooperation. ·METHODS: We enrolled 8 cases ( left eye in 3 cases, right eye in 5 cases ) of osteopetrosis patients referred from the Eye Hospital of Wenzhou Medical University during February 2012 to November 2016. Patients received ophthalmic examinations including visual acuity and diagnostic imaging tests in pre-operation and post -operation. All eyes were performed surgical optic nerve decompression through endoscopic approach in assist of image guidance system. We retrospectively analyzed the clinical data and surgical cooperation procedure of these cases and summarized nursing cooperation experience. ·RESULTS:The operations of 8 patients were completed successfully without massive hemorrhage. Mean visual acuity improved from pre-operation (2. 5±2. 1) to post-operation (3. 4±1. 9). Cerebrospinal fluid leakage occurred in 1 patient and was instantly repaired during the operation. We performed the nursing strategy as postural drainage, condition monitoring and conscious assessment intra-and post-operation. ·CONCLUSION: It is the critical for this kind of surgery that both circulating nurse's high-skilled cooperation to the connection and operation of the navigation system, to treat with complication during the surgery, and scrub nurse's sufficient preparation of surgical instruments and consumables, proficient equipment delivery, meticulous management, use and maintenance of equipment.

Objective To construct a nucleic acid vaccine expressing H.pylori HpaA and inter- leukin-2 gene and to identify the immunogenicity of the vaccine proteins in vitro and protection in vivo. Methods The H paA gene fragment was amplified by polymerase chain reaction(PCR) from the genomic DNA of the standard H.pylori strain 17874.Mouse interlukin(IL)-2 gene was amplified from pClneo- IL-2.The HpaA and IL-2 were cloned into pUCmT vector.After DNA sequences of the amplified HpaA gene and IL-2 were confirmed,both were cloned into the eukaryotic expression vector pIRES through a serial of enzyme digestion and ligation reactions.The recombinant plasmids were screened by PCR and restriction enzyme digestion.Then,recombinant pIRES-HpsA-IL-2 was transfected to COS-7 cells using Lipofectamine~(TM)2000.The immunogenicity of HpaA and IL-2 protein was detected by SDS- PAGE and Western blot.The recombinant plasmids were transformed to LB5000 and then to final host SL7207.The recombinant strains were passaged repeatedly.The mice were challenged with H.pylori after 4 weeks of inoculation of nucleic acid vaccine.H.pylori infection was detected by rapid urease test.Results The amplified HpaA gene fragment and IL-2 were confirmed by sequence analysis.The eukaryotic expression vector plRES and the pIRES-HpaA-IL-2 construction were confirmed by PCR and restriction digestion.The expressions of HpaA(30 000) and IL-2(14 000)protein by pIRES-HpaA-IL- 2 were detected by Western blot.The in vivo study showed that 75.0% and 58.4% of mice vaccinated by HpaA-IL-2 and HpaA,respectively,were protected anaigst H.pylory infection,which was signifi- cant different in comparison with PBS control (P

Objective To investigate the effect of ultrasound-and nerve stimulator-guided femoral nerve and lateral femoral cutaneous nerve block versus general anesthesia on knee joint surgery in elderly patients.Methods The 110 elderly patients with spinal anesthetic contraindication and undergoing lower extremity surgery from June 2014 to June 2015 were randomly divided into observation group (n =55) and control group (n =55).The observation group received both ultrasound-and nerve stimulator-guided femoral nerve and lateral femoral cutaneous nerve block,and the control group was given general anesthesia.Anesthesia procedure,sensory block onset time,changes in heart rate and mean artery pressure (MAP) after anesthesia,the total quantity of fluids infusion,dosage of vasopressor and hypotensor,adverse anesthetic reactions,anesthetic fees,anesthetic effect were recorded.Results Anesthetic preparation and practicing time had no difference between the two groups [(8.3 ± 1.7) min vs.(7.7 ± 1.2) min,(t =1.661,P=0.139)].The block onset time was longer in observation group than in control group [(10.3 ± 1.4) min vs.(3.2±0.6) min,t=50.180,P＜0.01].The changes in MAP had significant difference between the two groups [5 min after anesthesia:(89.24 ± 8.30) mmHg and (77.90 ± 8.05) mmHg;after operation:(96.60±8.03) mmHg and (106.22±8.88) mmHg;P＜0.05].There were significant differences in the fluid infusion quantity,dosage of vasopressor and hypotensor,adverse reactions during or after anesthesia,and anesthetic fees between the two groups [(1150.9± 231.6) ml vs.(1400.0±256.5) ml,(3.91±1.21) mg vs.(10.83±2.19)mg,(1.80±0.37) mg vs.(8.27±1.25)mg,3.6％ vs.18.2％,(1239.1±202.9) Yuan vs.(2307.2±205.6) Yuan,all P＜0.05].No significant difference was found in anesthesia effect between the two groups (P =0.198).Conclusions The ultrasound-and nerve stimulator-guided femoral nerve and lateral femoral cutaneous nerve block versus general anesthesia is more simple and safe for the knee joint surgery in elderly patients,with less complications,lower cost and higher satisfaction of patients.

Objective To study the imaging findings of unexpected splenic autotransplantation after splenectomy and to improve diagnostic accuracy of splenic autotransplantation.Methods The findings of 10 patients with splenic autotransplantation confirmed by histology in our hospital were retrospectively reviewed.In 7 patients CT and plain and dynamic enhanced MRI scanning were carried out,and in 2 of them 99mTc-DRBC scanning were also done.In 3 patients,plain and dynamic enhanced CT scannings were done.Results (1) Multiple lesions were detected in 7 patients and a single lesion in 3 patients.The masses were round and oval.The nodules were in the splenic fossa in 9 patients,in the pancreatic tail in 4 patients,in the right liver in 2 patients and in other of parts of the abdominal cavity in 2 patients.These nodules varied in size and 94.6％ showed a maximum diameter of less than 3 cm; (2) The findings on CT and MR:all the nodules were homogeneous with soft tissue density.There was no cystic degeneration,necrosis and calcification.In one patient with a nodule in the pancreatic tail,there was a slightly short T1 and short T2 signals.Other nodules showed long T1 and long T2 signals.All the signals from the nodules were homogeneous and their outlines were clear.In a patient with a nodule in the right liver,the blood supply came from the abdominal aorta.There was a surrounding thin layer of low-density ring which showed long T1 and long T2 signals.The enhanced features on CT and MR were similar.The nodules showed homogeneous or inhomogeneous enhancement in the arterial phase,with continuous homogeneous enhancement in the portal venous phase,with an obvious decline in the delayed phase;(3) The findings of 18 F-FDG PET:The nodules had obvious increase in FDG uptake.Conclusions In patients with a history of splenic trauma or splenectomy,abdominal nodules with multiple,homogeneous density or signal,clear outline,enhanced features similar to spleen,splenic autotransplantation should be considered.99mTc-DRBC scanning is helpful in the diagnosis of splenic autotransplantation.

Objective To evaluate the feasibility of dopamine D2 receptor imaging agent (s)-(-)-N-(1-allylpyrrolidine-2-N-methyl)-5-(3-18F)-2,3-dimethoxy Benzamide (18F-Fallypride) for targeting islet cell imaging.Methods (1) Cytology experiment:Islet cells of 15×103 cells/well were incubated with 3.70 kBq/well 18F-Fallypride for 1 h and the uptake rate of cells was calculated (cell counts/(supernatant counts + cell counts)× 100％).Under the same experiment conditions,6 inhibiting groups were administrated with different concentration of dopamine inhibitors droperidol (1.0× 10-6,4.0× 10-6,2.0× 10-5,1.0× 10-4,5.0× 10-4 and 1.0× 10-3 mol/L,respectively).After 30 min,3.70 kBq of 18F-Fallypride was added to each inhibiting group,and the inhibiting rate was calculated.(2) Autoradiography:18 normal ICR mice were divided into 6 groups.For group A,ICR mice were injected with 18F-Fallypride (55 ± 5) MBq/mice through tail vein.For the other 5 inhibiting groups (group B-F),ICR mice were injected with different doses of droperidol (0.2,0.4,0.6,0.8 and 1.0 mg/kg,respectively),and after 30 min 18F-Fallypride were injected through tail vein.Ten minutes later,pancreas of ICR mice was taken for preparation of tissue section autoradiography.The data were analyzed by one-way analysis of variance and the least significant difference t test.Results (1) The 18F-Fallypride uptake rate of control group was (18.40± 1.21) ％.The uptake rates of inhibiting groups were (16.11±1.37)％,(15.76±0.99)％,(13.90±1.02)％,(8.86±0.73)％,(7.26±0.62)％ and (6.92±0.58)％,respectively,which decreased with the decreasing concentration of droperidol (F=50.01,P＜0.01).When the concentration of droperidol was 1.0× 10-4 mol/L,the uptake rate reached the lowest with inhibiting rate of 51.85％.(2) The autoradiography showed that the pancreas gray scale value of group A was 1.21×106 digital light units (DLU)/mm2.The pancreas gray scale value of groups B to F decreased with increasing concentration of inhibitor:0.93× 106,0.77× 106,0.59× 106,0.32× 106 and 0.25×106 DLU/mm2,respectively.Conclusions 18F-Fallypride may specifically and efficiently bind to dopamine receptors of islet cells.It may be a potential tracer for islet cells imaging.

Objective To evaluate imaging features and clinical significance of anatomical variations of the dorsal pancreas.Methods CT and MR imaging data of 47 cases with variations of the dorsal pancreas were collected.Imaging characteristics of the dorsal pancreas were analysed.Results (1) Narrow dorsal pancreas:the agenesis of dorsal pancreas (7 cases) appeared as short pancreas,their length was (91.59 ± 22.39) mm.4 cases of pancreatic head volume increased,with tadpole-like retention of the pancreas,including 3 cases of annular pancreas.2 cases with polysplenia syndrome and congenital abscence of the hepatic segment of inferior vena cava.(2) Abnormaly enlarged dorsal pancreas:①the broadening of the pancreatic tail (n =18):the maximum diameter of pancreatic tail was (36.12 ± 6.59) mm,the pancreas was similar to the dumbbell-shaped.②Processes locally of pancreatic contour (n =13),which were local process at the ventral aspect of pancreas,the height was (15.72 ±2.65) mm,the width was (18.59 ± 3.64) mm,most often seen on the neck of the pancreas.(3) Dorsal pancreas related divisium:①Pancreas separated by fat spatium (n =7),the width was (3.51 ± 2.42),the deepness was (19.45 ± 5.84),it showed the crack-like fat density (signal) shadow,5 cases (5/7) located in the pancreatic body and tail,2 cases (2/7) located at the junction of ventral pancreas and dorsal pancreas.②The bifurcation of the pancereatic tail (n =3),limitations forked tail of the pancreas was dovetail-like performance.The maximum width diameter was (26.63 ± 1.75) mm,the bifurcation angle was (99.27 ± 30.73) degrees.Conclusions The developmental anomalies of dorsal pancreas can lead to a number of variations of pancreas,some of which can induce corresponding disease and be mistaken for neoplasm.