Objective To design a novel efficient CEA vaccine of fission epitopes with the vehicle of HPV16L1,and construct the recombinant baculovirus in Bac-to-Bac baculovirus expression system.Methods The recombinant baculovirus expression vector pFastBac1-V was constructed.HPV16L1 and CEA-PADRE synthesis primers were inserted into bacmid in E.coli DH10Bac with the help of Tn7 transposition system.Then the recombinant baculovirus was got by infecting insect cell sf9.Results HPV16L1 and CEA-PADRE synthesis primers that were cloned to the baculovirus transfer vector pFastBac1 were assessed by restriction analysis and direct sequencing.The recombinant baculovirus of the novel vaccine had been constructed.After infected sf9,the correct fragment was amplified by PCR.Conclusion The baculovirus of CEA vaccine of fussion epitopes with the vehicle of HPV16L1 was constructed.

Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

Objective:To develop non-immunized human phage display library.Methods:The total RNA of lymphocyte cells from peripheral blood of healthy voluntee was isolated and cDNA was synthesized,and the genes of heavy variable chain (VH) and light variable chain (V? and V?) were amplified by direct PCR and half-nested PCR.By overlapping extension PCR,the genes of VH and VL (V? and V?) were linked.The linked genes of single chain Fv fragment (scFv) were ligated with the vector pCANTAB-5E and then cloned into TG1 for the scFv library construction.Results:By direct PCR and half-nested PCR,42 VH fragments,16 V? and 18 V? fragments were obtained.The size of linked scFv library genes was 750 bp and the volume of constructed scFv library was 1.35?108.The results of BstN Ⅰ analysis of scFv genes from the phage library showed that fingerprint map of the selected scFvs was different.Conclusion:The developed phage library is diversity and can be used for selecting humanized scFv.

Objective A HPLC method was established to measure the contents of major nucleosides in Hyriopsis cumingii.Method Separation was performed on a Lichrosper-C_(18) column, pure water was used as mobile phase,the flow rate was 1.0mL min~(-1),the column temperature at 30℃and the detective walvelength at 254 nm.Results The contents of uracil,hypoxanthine, xanthine,uridine and thymine were 153.2～173.3?g?g~(-1),106.6～360.8?g?g~(-1),117.6～172.8?g?g~(-1),70.3～153.1?g?g~(-1) and 343.9～511.4?g?g~(-1),respectively. Conclusion This method showed good repeatability and flexibility.It could be used as a conventional method for determination of major nucleosides in H.cumingii.

0.05). (2) In the total 12 rabbits, all the cornea grafts survived without rejection reaction, corneal haze or corneal neovascularization. The fresh grafts got transparent after 2 months, and the dehydrated grafts got transparent after 6 months. Histological study demonstrated both fresh and dehydrated stroma grafts had fused with rabbits'corneal stroma very well without lymphocytes infiltrating. (3) As shown in histological observations, the reconstructed cat corneal endothelial tissue was similar to the nature tissue. Cultured cat endothelial cells connected tightly to each other and attached to the Descemet's membrane closely. CONCLUSION: Swine corneal stroma has low immunogenicity and satisfying biocompatibility,it is an ideal biological carrier for cornea reconstruction in vitro.

Objective To report the method for repair of the heel and ankle.Method On the basis of applied anatomy,the reversed island flap based on sural neurovascular pedicle has been designed for repair of heel and ankle defects.Results The flaps have been used in 14 cases,the maximum size 9 cm×15 cm,and all flaps were survival compeletely except partial necrosis in one flap.Conclusion The blood supply of the flap is good.The length of pedicle can reach 12 cm.It is an effective method for repair of heel and ankle defects.

han that in the control one (t=3.94, P<0.01).Conclusion Social skills training can effectively improve subjective satisfactions of the long-terra hospitalized patients with chronic schizophrenia.

OBJECTIVETo explore the clinical anesthesia value of transcutaneous acupoint electrical stimulation (TAES) combined with general intravenous anesthesia in endoscopic bilateral thyroidectomy patients.

METHODSTotally 60 patients who underwent endoscopic bilateral thyroidectomy were equally randomly assigned to 2 groups, the treatment group and the control group, 30 in each group. Patients in the treatment group received TAES combined general intravenous anesthesia, while those in the control group received total intravenous anesthesia. Anesthesia was maintained by target controlled infusion of propofolum combined constant speed infusion of remifentanil in the two groups. TAES was maintained from 30 min before anesthesia induction to the end of endoscopic thyroidectomy at bilateral Hegu (L14) and Neiguan (PC6). The mean artery pressure (MAP) and heart rate (HR) were recorded at different time points of anesthesia, i.e., immediately after entry into the operating room (TO), immediately after intubation (T1), 5 min after intubation (T2), 5 min before incision (T3), 5 min after incision (T4), 30 min after inflation (T5), at the end of surgery (T6), 5 min before extubation (T7), immediately after extubation 0 (T8), and 5 min after extubation (T9). The concentration of IL-6 and TNF-alpha were measured at TO, T3, T5, and T6. The target concentration of propofol was also recorded at T3, T4, and T5.

RESULTSThere was no statistical difference in the operation time between the two groups (P >0.05). Compared with TO in the same group, HR at T3-T4 decreased and increased at T8-T9, and MAP increased at T7-T9 in the treatment group; HR decreased at T3 and increased at T7-T9, MAP increased at T1, T5, T7-T9, and MAP decreased at T2-T3 in the control group. IL-6 increased at T5-T6, while TNF-alpha decreased at T6 in the two groups (P <0.01,P <0.05). Compared with the control group at the same time point, HR decreased at T6-T9, MAP decreased at T1, T4, T5, T7-T9, MAP increased at T3, and IL-6 decreased at T5-T6 in the treatment group (P <0. 05). The concentration and the total amount of propofol were significantly lower in the treatment group than in the control group (P <0.01,P <0.05).

CONCLUSIONSTAES could maintain the hemodynamics more stably and inhibit the stress response in endoscopic thyroidectomy. It also reduce the dosage of anesthetics and improve the safety of anesthesia.

Acupuncture Points ; Anesthesia, General ; Anesthesia, Intravenous ; Electric Stimulation ; methods ; Endoscopy ; methods ; Heart Rate ; Hemodynamics ; Humans ; Interleukin-6 ; Piperidines ; Propofol ; Thyroidectomy ; methods ; Transcutaneous Electric Nerve Stimulation ; Tumor Necrosis Factor-alpha

Objective To evaluate the diagnostic value of endoscopic ultrasonography (EUS), and explore the efficacy of endoscopic treatment in patients with esophageal submucosal tumor. Methods Sixty-eight patients with esophageal submucosal tumor were selected, and the tumor was derived from the muscularis mucosa and submucosa according to the common endoscope and endoscopic ultrasonography detection. Endoscopic mucosal resection (EMR) was applied to remove submucosal tumor with diameter less than 1.0 cm, endoscopic piecemeal mucosal resection (EPMR) or endoscopic submucosal dissection (ESD) was applied to remove submucosal tumor with diameter 1.1 - 1.5 cm, and ESD was applied to remove submucosal tumor bigger than 1.5 cm. Samples were examined by pathology after treatment. Results Tumors in all the patients were completely removed, and the tumor diameter was 0.6-2.3 cm. Forty-one cases were treated with EMR, 9 cases were treated with EPMR and 18 cases were treated with ESD. Four patients had intra-operative bleeding that was stopped by electrocoagulation hemostasis. No perforation occurred in all cases. Postoperative pathology revealed 43 cases had leiomyoma, 23 cases had interstitialoma, and 2 cases had lipoma. Patients were reviewed by gastroscope 3 months after operation. The white scars formed in all patients, and there was no residue or recurrence. Conclusions Different origin layers and property of esophageal submucosal tumor can be diagnosed accurately by EUS, and endoscopic therapy (EMR, EPMR and ESD) is an effective treatment for submucosal tumor from muscularis mucosa and submucosa. Endoscopic therapy is safe and effective. It provides sufficient pathological information.

Objective To determine the function and clinical significance of OMOM capsule endos-copy. Methods To review the history and outcomes of 65 patients underwent capsule endoscopy from Oct. to Dec. 2004. Results OMOM capsule endoscopy failed to pass the pylorus within the effective working time in one case. The significant pathological findings were revealed in 47 patients among 64 patients (73. 4% ). Inflammatory small bowel diseases in 21 patients including Crohn's disease in 3 patients, small intestinal pol-yps in 8 patients including one Peutz-Jeghers syndrome, angiodysplasia in 9, diverticula in 3, hookworm in 3, submucosal tumor in 3 including one myosarcoma finally diagnosed by operation. OMOM capsule endosco-pies have working time 473 min (360-630) averagely. The duration from oral to anus was 1723 min (690-2370 min) averagely. Conclusion The capsule endoscopy is a highly useful technique in detecting small in-testinal diseases. It can be recommended as part of the routine work-up in patients with obscure bleeding.