Objective:To produce monoclonal antibodies(McAb) against aspergillus fumigatus and to establish rapid assay for the measurement of aspergillus fumigatus antigen.Methods:Recombinant galactomannoprotein of aspergillus fumigatus(AFMP1) was used to immune BALB/c mice.Monoclonal antibodies against AFMP1 were produced from hybridoma.Results:Three hybridomas producing antibodies against AFMP1 were obtained.IgG isotypes of three McAb were IgG1,IgG2a and IgG2b.The affinity constants(K) were 1.2?10 10 ,4.56?10 9 and 1.81?10 10 mol/L.The antibodies were proved to be specific for aspergillus fumigatus by Western blot and recognized different epitopes on AFMP1 by the additivity assay.An sandwich enzyme-linked immunosorbent assay(ELISA) to detect AFMP1 was established to produce standard curve which showed linearity between 0.1～60.0 ng/ml with a sensitivity of 0.1 ng/ml.Conclusion:These results show three hybridomas producing high specificity and affinity monoclonal antibodies against AFMP1 and can provide for rapid assay for the measurement of aspergillus fumigatus antigen.

Objective:To investigate the in vitro the anti-prostate cancer effect of a novel polypeptide,APP216,so as to provide a basis for development of new drug for treatment of prostate cancer.Methods:The polypeptide drug included the amino sequences of BH3,K237 and DG2 domain and the peptide that could be digested by PSA.The anti-prostate cancer effects of the polypeptide prodrug on prostate cancer cell line LNCaP,22RV1(secreting PSA)and PC3,DU145(secreting no PSA)were determined by MTT test and Hoechst 33258 staining.Results:MTT test revealed that the surviving rates of LNCaP and 22PV1 cells were respectively 22% and 34% 48 h after APP216(270 ?g/ml)treatment,and 9.8% and 8.2% 72 h after APP216(270 ?g/ml)treatment.The surviving rates of PC3 and DU145 cells were respectively 90% and 95% 48 h after APP216(270 ?g/ml)treatment,and 87% and 92% 72 h after APP216(270 ?g/ml)treatment.Hoechst 33258 staining showed the typical features of cell apoptosis:cell shrinkage,chromatin condensation and hypodiploid genomic DNA content in LNCaP and PC3m cells.Flow cytometry showed an apotosis rate of 36.26% 48 h after APP216(270 ?g/ml)treatment in LNCaP cells,and of 1.63% after 48 h APP216(270 ?g/ml)treatment in PC3m cells.Conclusion:APP216 has a satisfactory in vitro cytotoxicity on human PSA-secreting prostate cancer cells and can induce tumor cell apoptosis,but not on non-PSA secreting prostate cancer cells,indicating APP216 polypeptide can be specifically digested by endonuclease enzyme.Meanwhile,BH3 domain can be transferred into cells and induce apoptosis through HIV-TAT.

Objective To report our usage of a combined flap which is constituted of bilateral hallux nails, skins, bones to reconstruct a finger, and to introduce the method and outcome of this way. Methods Combine two halves of halluxes harvested from both feet to reform a fabricated finger and then transplant it to the finger stump to reconstruct the defect part of the finger. Plantar flaps or some other flaps near the donor sites were transposed to cover them. From June 2003 to June 2009, a total of 20 fingers (20 cases) which had defect degrees range from I to Ⅲunderwent reconstruction surgeries in this way. Results All the 20 fingers transplanted survived completely. Follow-ups 1 to 5 years after each surgery: all the fabricated fingers had very realistic configurations. The MP joints of the reconstructed thrumbs got to the normal range of motion, and the other reconstructed fingers' total ROM were 203 degree on average. All the reconstructed fingers had the sensation function above S3,and their two-point discriminations ranged from 6mm to 10mm. Both halluxes of each case were conserved major parts of nails and had nice, symmetric appearances. All the flaps for the donor halluxes survived completely, and none of the cases showed pains, ulcers or abrasions of their feet. All the cases showed normal gaits during follow-ups. Conclusion The combined flap by bilateral hallux nails, skins, bones is an ideal alteration for finger defect reconstruction for the important advantages of realistic configuration as well as minor destructions to donor sites.

Objective To observe the therapeutic effect of tramadol combined with fentanyl in patient controlled intravenous analgesia after nucleotomy of intervertebral disc.Methods 60 patients under general anesthesia,scheduled for nucleotomy of intervertebral disc,were randomly divided into three group(n=20):group tramadol(group T),group fentanyl(group F) and group tramadol+fentanyl(group TF).All patients were received patient controlled intravenous analgesia by loading dose+background dose+PCA bolus dose.The BP,HR,RR,VAS at 6,12,24,48h of postoperation were observed.The sedation score in 24h,average effective-press times and drug dose in 48h of postoperation were recorded.The incidence of nausea and vomiting during 24h of postoperation were observed.Results It had no significantly difference of BP,HR,RR,VAS among three groups.The average effective-press times of group TF (8?2) were significantly less than those of group T (14?3) and group F (12?3).The drug dose of 48h was tramadol (17.9?1.5)mg/kg(group T),fentanyl (16.8?1.5)?g/kg(group F) and tramadol (11.5?0.7)mg/kg+fentanyl (9.1?0.7)?g/kg(group TF).The drug dose of group TF showed significant difference compared with others.The sedation score in group F was the highest,it had significant difference compared with the other two group.The frequency of nausea and vomiting in group T was significantly more than that in others.Conclusion Tramadol or fentanyl gived alone can afford the same satisfied analgesic effect on nucleotomy of intervertebral disc.But there is high nausea-vomiting frequency in tramadol and strong sedation frequency in fentanyl.Tramadol combined with a little opioid fentanyl can afford more satisfying analgesic effect,and is provided with advantage in less drug and side-effect.

e advantages as those of digital artery flap, but also has proper digital artery and nerve being untouched.

BACKGROUND: A lot of important organs are worthless for clinical application because they are hard to store for a long time. In addition, tissues or organs which are dealt with cryopreservation also attack ischemia/reperfusion injury with the recovery of blood flow; especially, skeletal muscle is the most involved tissue.OBJECTIVE: To observe the protective influence of edaravone on cellular membrane and mitochondria of replanted rat extremities following ischemia/reperfusion injury due to cryopreservation and rewarming.DESIGN: Randomized contrast animal study.SETTING: Basic Medical College of Southern Medical University; Department of Hand and Foot Surgery, Shandong Provincial Hospital.MATERIALS: The experiment was carried out in the Cryopreservation Laboratory, Shandong Provincial Hospital from April to November 2006. A total of 36 healthy adult male Wistar rats were provided by Experimental Animal Center of Medical College of Shandong University. All rats were randomly divided into control group, cryopreservation group and edaravone group with 12 in each group.METHODS: Femoral artery and vein of rats in control group were exposured, but extremities were not blocked. Rats in other two groups were used to establish ischemia/reperfusion injury models of replanted extremities. Before cryopreservation, their right hindlimbs were cut off and maintained in liquid nitrogen container for 1 month. After the operation mentioned above, the broken limbs were rewarmed, perfused with routine eluant and replanted. Four hours later, blood supply of extremities was recirculated and the samples were selected. Eluant in edaravone group contained 0.5 mg/kg edaravone. Samples of skeletal muscle were selected at the same time point to establish cellular membrane and extract mitochondria. Furthermore, fluorescence polarization of cellular membrane (reflecting liquidity in cellular membrane lipid area), malondialdehyde (MDA) content of mitochondria, superoxide dismutase (SOD) activity and respiratory controlling rate were measured; meanwhile, mitochondrial ultrastructure of skeletal muscle was observed under transmission electron microscope.MAIN OUTCOME MEASURES:①Fluorescence polarization of cellular membrane, MDA content of mitochondria, SOD activity and respiratory controlling rate of skeletal muscle; ②mitochondrial ultrastructure of skeletal muscle.RESULTS: All 36 rats were involved in the final analysis without any loss. ①SOD activity and respiratory controlling rate of mitochondria in skeletal muscle: The values of these two items were higher in edaravone group that those in cryopreservation group (P＜0.05).②Fluorescence polarization of cellular membrane and MDA content of mitochondria in skeletal muscle: The values of these two items were lower in edaravone group than those in cryopreservation group (P＜0.05). ③Mitochondrial ultrastructure of skeletal muscle: Injured degree of skeletal muscle was milder in edaravone group than that in cryopreservation group.CONCLUSION: Edaravone can relieve ischemia/reperfusion injury of skeletal muscle and protect cellular membrane and mitochondria of skeletal muscle. Its mechanism may be related to directly inhibiting hydroxy free radicals, increasing SOD activity of skeletal muscle, reducing generation of MDA and promoting normal oxidative phosphorylation.

Objective To evaluate the design formulas of the optimal location for splitting pectoralis major in terms of making type Ⅰ or Ⅱ dual-plane implant pocket.Methods Sixty-five patients with micromastia were collected.Breasts were divided into two types according to the soft-tissue pinch thickness of the lower pole:type Ⅰ (thickness＞2 cm;34 cases) and type Ⅱ (thickness ≤2 cm;31 cases).The optimal levels at which the pectoralis major (PM) was severed were 3/4 or 2/4 of the distance of new inframammary fold for type Ⅰ or Ⅱ dual-plane pockets,respectively.All patients completed the pre-and post-operative BREAST-Q augmentation modules before and 6 months after surgery.The scores were changed into hundred-mark system by QScore software.Results The recovery processes were well-off.The breasts contour was good.Patients reported higher scores of satisfaction with breasts,psychosocial well-being and sexual well-being after surgery than before surgery (62.0±8.9 vs 27.9± 4.3,65.0± 17.2 vs 17.4±8.3,60.5± 14.2 vs 30.3±5.5,P＜0.05).The mean satisfaction score for the overall outcome was 90.6±5.4.However,there was no significant difference in physical well-being between before operation and aftre operation (85.3±9.5 vs 84.7± 10.6).No complications such as severe capsular contracture,or displacement occurred.Conclusions The design formulas make the determining procedure of the optimal location for pectoralis major splitting for two types dual-plane implant pockets easier and more exactly.Our modified design method can provide the implant with the optimal soft tissue coverage,and bring desired and stable breast aesthetic outcomes.The higher satisfaction and quality of life reported by patients indicate that the formulas are feasible and worth to recommend.

Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

Objective: To examine the influencing factors of the efficiency of county-level centers for disease control and prevention ( CDCs) in China. Methods:458 county-level CDCs were selected based on a systematic sam-pling method. Multilevel modeling was used to analyze the region-level and institution-level influencing factors affect-ing the efficiency of CDCs. Results: It was found that the region ( province) is associated with the efficiency of a CDC. The region-level factor of population density exhibited a significant influence, while the institution-level factors such as the proportion of health technicians, service income and CDC laboratories per capita also had an influence on overall efficiency. Conclusions: Both the region-level and institution-level determinants influence efficiency. Multi-level modeling can help researchers gain a comprehensive understanding of the influencing factors that affect the CDC efficiency.

Background and Purpose:Lung cancer is the most malignant tumour in the world.Its incidence is growing and NSCLC is predominent(80%) in lung cancer.Most patients with lung cancer were diagnosed in late stages.The tumour could be shrunk by neoadjuvant chemotherapy when the case with stage Ⅲ NSCLC was considered not possible for radical operated neoadjuvant chemotherapy may lead to the following,operation could be improved,micrometastasis could be annihilated and survival could be extended.Objective of this paper was to analyse the prognostic factors for survival in patients treated by surgery and chemotherapy for NSCLC.Methods:98 cases of neoadjuvant chemotherapy combined with surgery for NSCLC,stageⅠ~Ⅲ,were collected retrospectively in our hospital from 1995 to 1997.35 cases were stageⅠ.21 cases were stage Ⅱ.42 cases were stage Ⅲ.83 cases had 1 cycle of chemotherapy pre-operatively.15 cases had 2 cycles chemotherapy pre-operatively.Regimes of chemotherapy were MVP,MOP and MAP et al.Response rate(RR) of chemotherapy was:45 cases had partial response(PR) and 53 cases were stable disease(SD).Operative mode was lobectomy and pneumectomy with lymph nodes dissection.Pathologic type was squamous,adeno,adeno-squamous and others.All the patients were treated by chemotherapy for two or three cycles after surgery except for the patients in stageⅠin 1996~1997.After being followed-up for more than 5 years,data were examined using life table,KaplanMeier method,Log Rank statistic and Cox-mantel test.The possible factors affecting survival were tested with univariate and multivariate analysis.Results:The median followed-up time of 98 cases for NSCLC was 41.2 months.36 cases were alive.62 cases were dead.The 1-,3-,5-year survival rate of 98 cases for NSCLC was 88.78%、49.63% and 18.46%.The 5-year survival rates of stageⅠ、ⅡandⅢ were 33.23%、20.26% and 5.52% respectively(P=0.0002).The 5-year survival rates of N_(0)、N_(1)、N_(2) were 35.49%、19.08% and 4.90% respectively(P=0.0004).In the 98 cases of NSCLC,better prognosis was correlated with earlier stage.The prognosis was better if the period from last chemotherapy before operation to operation was no more than 1 month. The prognosis of lobectomy,lung hila activity,thorax lymph nodes negativity and squamous cancer was better.The prognosis was poorer if the tumor had invaded big vessels,viscera,chest wall,pericardium and quantity bleeding during≥400ml.The prognosis was better if the tumor was fibrotic.The prognosis of 2 cycles of chemotherapy pre-operatively might be better than 1 cycle.The prognosis of tumor necrosis was poorer and the prognosis of chemotherapy post-operatively was better.Conclusions:The main prognostic factors affecting survival in patients treated by surgery and chemotherapy for NSCLC was stage,the period from last chemotherapy before operation to operation,operation mode,lung hila activity,thorax lymph nodes,site of tumor invasion,bleeding quantity,pathologic type,tumor fibrosis and necrotis,cycles chemotherapy pro-operation and chemotherapy post-operation.