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 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

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 analyze the operation methodes and effects of cataract complicated with acute angle-closure glaucoma . Methods Phacoemusification combined with intraocular lens implantation and phacoemusification ,intraocular lens implantation combined with trabeculectomy were choosed to be performed on 40 eyes(38 patients) which were diagnosed with cataract compli-cated combined with acute angle-closure glaucoma ,according to the intraocular pressure and the opening level of the angle of anteri-or chamber of patients .All cases were been followed-up from 6-24 months after the operation .Results For postoperative correc-ted vision ,20 eyes were greater than 0 .6 ,17 eyes were between 0 .3-0 .6 and 3 eyes were less than 0 .3;For postoerative intraocu-lar pressure ,38 eyes were in normal range ,2 eyes were in normal range with drug control .Conclusion Phacoemusification combined with intraocular lens implantation prefer to be choosed for cataract complicated with acute angle-closure glaucoma with opening lev-el of the angle of anterior chamber greater than the half .Phacoemusification ,intraocular lens implantation combined with trabeculec-tomy prefer to be choosed for cataract complicated with acute angle-closure glaucoma with opening level of the angle of anterior chamber less than the half .Choosing proper operation methodes for the patients who were diagnosed with cataract complicated with acute angle-closure glaucoma ,according to patients′situation ,benefit to improve patients′vision and control intraocular pressure .

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 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

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 present the changing of the parameters of orbital volume after endoscopic transethmoid medial orbital wall decompression combined with the endoscopic transethmoid intraconal fat-removal orbital decompression in thyroid associated ophthalmopathy (TAO).Methods A retrospective chart was reviewed in 11 patients (20 eyes) receiving orbital decompression for the treatment of exophthalmos secondary to TAO from September 2014 to August 2015. All patients diagnosed TAO were in stable and inactive phase at least for 6 months. High-resolution computed tomography (HRCT) scan were performed in all patients before and 3-month after surgery. CT scan of orbit and computer-aided measurement software were used to measure the exophthalmos. Changing of the parameters of orbital volume were recorded for analysis its relationship with the amount of proptosis reduction.Results The exophthalmos was signiifcantly decreased after surgery. Medial rectus volume, fat volume and orbital volume were larger postoperatively. The changing of rectus volume and fat volume both had negative correlationship with the amount of proptosis.Conclusions The thickening of medial rectus 3-month postoperative may due to the removing of medial wall of orbit during the surgery. The stimulation of the surgery may be another reason. The changing of rectus volume and fat volume both had negative correlationship with the amount of proptosis.

Objective To evaluate the feasibility, safety and efficacy of CT-guided radiofrequency ablation in treating osteoid osteoma located at femoral neck. Methods Six patients with osteoid osteomas in the femoral neck received CT-guided percutaneous radiofrequency ablation. In all patients the main complaint was pain at the hip, and the course of disease varied from one month to 2 years, with an average of 8 months. Under spinal anesthesia the surgery was performed. With the help of CT guidance , a 3.5 to 4.0 mm coaxial drill system was inserted into the nidus, and an osseous access was established, then the bone biopsy needle was used to obtain specimens for pathological examination. Subsequently, a 1.5 to 2.0 cm active tip was introduced through a non-cooled radiofrequency needle into the nidus. Radiofrequency ablation was performed with the therapeutic temperature of 90℃, lasting for 6 minutes. The pain visual analogue scale (VAS) was used to evaluate the clinical effectiveness. The postoperative MRI findings were compared with the preoperative ones. Results Three days after the treatment, different degrees of pain relief was obtained in all patients, and all patients could get out of bed and walked around in one week. Postoperative VSA was significantly decreased (P<0.01). No severe complications occurred during and after the procedure. And no recurrence was seen during the follow-up period. Conclusion For the treatment of osteoid osteoma located at femoral neck, CT-guided radiofrequency ablation is a safe and effective minimally invasive treatment with fewer complications and satisfactory clinical results.