Objective:To construct eukaryotic inducible expression plasmids pTRE-Tight-CFP-CD11b and pTRE-Tight-YFP-CD18 using Tet-On gene expression system and co-transfect them into Chinese hamster ovary(CHO) cells,so as to achieve the inducible expression of Mac-1-FP.Methods: The eukaryotic expression vectors pTRE-Tight-CFP-CN11b and pTRE-Tight-YFP-CD18 were constructed by recombinant DNA technique.The 2 vectors were co-transfected into CHO cells using liposome to fuse CD11b and CD18: the 2 subunits of Mac-1. Fluorescence microscope was used to observe the cyan fluorescence and the yellow fluorescence of Mac-1-FP.The influence of different levels of Dox(0,0.01, 0.1,0.5,1,2 ?g/ml) on expression levels of CD11b and CD18 in CHO cells was analyzed by RT-PCR and fluorescence intensity analysis.The adhesive rate of CHO-Mac-1-FP cells with ligand ICAM-1 was analyzed before and after PMA(1 ?g/ml) stimulation.Results: The recombinant plasmids of pTRE-Tight-CFP-CD11b and pTRE-Tight-YFP-CD18 were successfully constructed.The cyan and yellow fluorescences were observed in co-transfected CHO cells under fluorescence microscope.The fluorescence intensity of the cells was increased with the increase of Dox concentration.RT-PCR analysis demonstrated that the CD11b and CD18 mRNA increased with the increase of Dox concentration.The adhesive rate of CHO-Mac-1-FP cells with ICAM-1 was increased after PMA stimulation(peaked at 2 h and 4 h after stimulation and decreased afterwards).Conclusion: This study achieves the inducible expression of Mac-1-FP in CHO cells.And Mac-1-FP,like widetype Mac-1,exhibits adhesive activity with ligand ICAM-1,which lays a foundation for studying the consisting dimmer,clustering,conformation and affinity of the ligands of Mac-1 using single molecule spectroscopy and fluorescence resonance energy transfer technique in living cells.

Objective To analyze the laboratory determination results of hand-foot-mouth disease(HFMD)and explore its sig-nificance.Methods Pharyngeal swab specimens were collected from 502 children with HFMD(severe case:104 cases and mild case:398 cases).Specific RNA of enterovirus primer(EV),enterovirus71 (EV71)and coxsackievirus A16(CA16)were detected by real-time fluorescence polymerase chain reaction(RT-PCR).Simultaneously,white blood cell (WBC),direct bilirubin(DBIL),alanine transarninase(ALT),creatine kinase MB isoenzyme(CK-MB)and lactic dehydrogenase (LDH )were determined,and the results were analyzed.Results Among the 502 children,384 cases were positive for EV,262 cases were positive for EV71,and 39 cases were positive for CA16.The positive rates were 76.49%,52.19% and 7.76%,respectively.In the 104 HFMD severe cases,EV71 positive rate was 92.31%(96/104).There were clearly more boys than girls.The age distribution was mainly in the group under 3-year-old(402 cases,80.08%).In the group under 3-year-old,there were 97 severe cases(93.27%),and the EV71 positive rate was 92.78%(90/97).Serum levels of WBC,DBIL,ALT,CK-MB and LDH were higher in severe HFMD group than in mild group(P <0.05).Conclusion The HFMD occurred mostly at the children under 3-year-old.The HFMD appeared is mainly related to the EV infection,and EV71 is the main pathogen causing the severe cases of HFMD.

Objective To investigate the influential factors of brachial-ankle artery pulse wave velocity (baPWV) in healthy individuals.Methods Total 716 healthy subjects including 503 males and 213 females,who underwent health check up in 2010,were enrolled in the study Bilateral baPWV was examined in all subjects and multiple factor linear regression was applied to evaluate the correlations of baPWV with sex,age,body mass index,arterial blood pressure,ankle-brachial index and biochemical variables.Results The mean baPWV of 716 healthy subjecs was ( 1224 ± 162) cm/s,and there was significant difference between men and women [ ( 1257 ± 159) cm/s vs.( 1144 ± 143) cm/s].There was a significant linear correlation of baPWV with sex,age,mean arterial blood pressure and plasma homocysteine concentration ; and the linear regression equation was:mean PWV =6.635 × age + 7.285 × mean arterial blood pressure + 2.682 × homocysteine concentration - 64.334 × sex + 347.500 ( sex:man =0,woman =1 ).Conclusion Age,sex,mean arterial blood pressure and plasma homocysteine concentration are independent influential factors of baPWV.

To study the effects of puerarin on the aromatase P450 (P450(arom)) mRNA expression and the effects of low-dose puerarin on transcription factors of the P450(arom) gene (P II) 5'-flanking region.

Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.

Objective To evaluate the efficiency and safety of hair removal in different areas with Lightsheer Duet diode laser and to compare clinical effect and safety in axillae hair removal between the two models,high-speed and standard model.Methods Lightsheer Duet diode laser was used to remove hairs from different areas in 200 cases,for 4-8 weeks treatment 1 time,4 times for 1course.The curative effect and adverse reaction were observed after the treatment.Thirty axillae hair removal subjects were chosen randamly and received treatments using the different models,high-speed hand piece on one side and the standard hand piece on the other side.Hair removal efficiency and the immediate pain were compared and evulated on each subject.Results The hair was significantly reduced after 4 times treatments in 200 cases.The total effective rate was 95.54 ％ and positively related with the treatment times.Pigmentation was found in 3 cases,but it disappeared after 1-3 months.The total effective rate was 100 ％ after the 3 times treatments in axillae hair removal subjects both using the high-speed model or the standard model in 30 cases.There was no statistical difference between the two models except less pain in the high-speed model.Conclusions Lightsheer Duet diode laser is a safe and efficient method in hair removal.The high-speed model has more advantage on speed and causing pain.

Objective To compare the efficacy of transumbilical-ultrafine gastroscope sympathectomy for severe palmar hyperhidrosis using two distinct levels of T3 and T4.Methods A total of 84 cases with severe primary hyperhidrosis were randomly allocated to undergo either T3 sympathectomy treatment (T3 group, n =42) or T4 sympathectomy treatment (T4 group, n =42) with no difference between the two groups.The operation time, postoperative hospital stay, patient's hands hyperhidrosis, axillary hyperhidrosis,complications were recorded at follow-up in 1,3,6,12 months.Results Operation on 84 patients were successful with mean operative time of T3 group being 55.02 ± 10.61 min and T4 group being 55.36 ± 10.51 min(P ＞ 0.05).The mean postoperative hospital stay were both 1.5 days.Patients were followed up for diaphragmatic hernia, umbilical hernia, Horner's syndrome and other serious complications for 12 months.No postoperative recurrence of palmar hyperhidrosis, severe compensatory sweating occurred in either group.The number of improved patients in T3 group's palmar hyperhidrosis,axillary hyperhidrosis, and foot hyperhidrosis were :42/42 cases, 10/16 cases, 21/29 cases, while those in T4 group were: 42/42 cases,16/17 cases 18/28 cases.Axillary hyperhidrosis improved to a larger extent in T4 group than in T3 group (P ＜0.05).There were 16(15 mild and 1 moderate)compensatory sweating in T3 group and 7(6 mild and 1 moderate) compensatory sweating in T4 group (P ＜ 0.05).Conclusion T3 and T4 thoracic sympathectomy using transumbilical-ultrafine gastroscope for primary palmar hyperhidrosis are safe, effective, and feasible.T4 sympathectomy is more effective in improving axillary hyperhidrosis than T3 and shows lower occurrence of postoperative compensatory hyperhidrosis.

Objective To evaluate the technique of finger palpation in thoracoscopic localization in patients with pulmonary nodules,and to summarize its technical details,especially with exploit of chest computed tomography (CT) facilitating it.Methods 95 patients with total amount of 109 pulmonary nodes 20 mm or smaller in size shown with lung window of CT,were reviewed.They were located subpleurally,with a median depth of 8.2 mm and a median size of 10.0 mm.The value of their depth over their size (D/d value) could be used as the extent of localizing difficulty.Each node had its own radiographic fealures for being localized,which was built preoperatively.Under thoracoscopic vision,nodules were finger-palpated by index finger via the 4th or 5th intercostal space on anterior axillary line,followed by wedgectomy or lobectomy for instant histopathological diagnosis to further decide the final surgical type.The distance between the nodule and the origin of segmental bronchus (L value) were also calculated out,as it might be relevant to the way the nodule could be biopsied.Results All nodules were successfully localized and resected for biopsy goal,105 by wedgectomy,4 by lobectomy.After intraoperative diagnosis was made by the pathologist,VATS lobectomy and lymph node dissection were further performed in 55 patients.L value of 4 cases being biopsied by lobectomy ranged from 18.3 to 30.3 mm,averaging 26.1 mm.Conclusion Finger palpation is viable in any cases of pulmonary nodules.Detailed reference of CT digital information,and enough detachment of mediastinal pleura,can greatly facilitate thoracoscopic localization by finger palpation.Lobectomy or segementectomy is preferable when L value is less than 30 mm.

Objective To evaluate the safety and feasibility of endoscopic transumbilical thoracic sympathectomy.Methods The technique was performed in 4 swine models.Under general anesthesia,a newly developed long abdominal trocar (60 cm in lengths) was placed through the umbilicus,which was also used to establish pneumoperitoneum.After insertion of gastroscope through this trocar,two small incisions of diagrams were created by needle-knife,through which the gastroscope was sent into the thoracic cavity to ablate the T3 ganglia by Hot biopsy forceps under One-lung ventilation.The animals were killed at the completion of the procedure.Results This procedure was completed in four pigs.The sympathetic chain was successfully ablated in all swine,as confirmed by gross surgical pathology and histology,the mean operative time was (81.7 ±15.4) minutes.There was no major bleeding.All bilateral T3 sympathetic chains were successfully ablated in the 4 swines as confirmed by gross surgical pathology and histology.There was no severe bleeding and damage to peripheral organs during the operation in any of the cases,and no prominent pneumothorax was revealed by postoperative chest X-ray radiography.Conclusion Endoscopic transumbilical thoracic sympathectomy is safe,feasible and effective with good cosmetic result.

Objective To explore a novel long-circulating dual-receptor targeting and dual-modal molecular probe and investigate its physicochemical properties and targeting effect on breast cancer cells in vitro. Methods Dual-receptor targeting and dual-modal molecular probe RGD@BBN-lipo(QDs)-SPIO was synthesized in the following steps: long-circulating liposome was prepared by film dispersion method;water-soluble superparamagnetic iron oxide (SPIO) nanoparticles and Quantum dots (QDs) were loaded in the hydrophilic and hydrophobic layer of liposome, respectively;RGD and BBN polypeptides were coupled on the former functional magnetic/fluorescent liposomes. Stability of the probe in different physiological solutions was investigated. Transmission electron microscopy (TEM) and particle size analyzer were used to measure nanoparticle sizes and the Zeta potential. Characterization of RGD and BBN was investigated through 1H-NMR and elemental analysis. The MRI T2 relaxivities (1/T2) of RGD@BBN-lipo(QDs)-SPIO was measured through T2 map scanning on 3.0 T MR system. HUV-EC-C cells were used for assessment of cells viability by MTS assay. Prussian blue staining and fluorescence imaging were carried out to determine the targeted breast cellular uptake of RGD@BBN-lipo(QDs)-SPIO nanoparticles. Results The targeting magnetic/fluorescent dual-model molecular probes appeared spherical or para-spherical,with a mean diameter of(118.2±3.9)nm,Zeta potential of (-24.78±1.68) mV,MR T2 magnetic relaxation rate of 0.498 1× 106 M-1 · s-1.RGD and BBN polypeptides were successfully coupled on the former functionally magnetic/fluorescent liposomes with the bind rates of 33.05%and 45.06%, respectively. There was low cytotoxity of the molecular probe on human umbilical vein endothelical cells(HUV-EC-C)by MTS study. Prussian blue staining and fluorescence imaging studies showed that the RGD@BBN-lipo(QDs)-SPIO nanoparticles could target any αvβ3 or gastrin releasing peptide receptor overexpression breast cancer. Conclusions RGD@BBN-lipo(QDs)-SPIO is a novel long-circulating dual-receptor targeting and dual-modal molecular probe and has excellent physicochemical properties and stability, high T2 relaxivities and strong targeting effect on cancer cells and has laid a solid foundation for early diagnosis of breast cancer.