Objective To search for the gene encoding specific antigen of Ancylostoma caninum that induces host′s protective immunity. \ Methods\ A lambda ZAPⅡ cDNA library of A.canium was screened with sera from dogs \{immunized\} subcutaneously with hookworm larvae(L3). After sequencing, insert gene (AcAg) from positive clones was \{ligated\} into PUC18 and PET28C. Recombinant pET28C plasmid was induced to expressed protein in the E.coli BL21. The \{characteristic\} of recombinant protein is analyzed by SDS\|PAGE and Western bloting assay. \ Results \ Five positive \{clones\} were obtained, and proved to be the same. The insert gene (AcAg) in pET28C vector expressed a recombinant \{protein\} of about 43 kDa. Using Western bloting assay, this protein was recognized by the sera from dog immunized with \{Ancylostoma\} caninum third\|stage infected larvae and used for screening library. \ Conclusion \ The AcAg, which \{exhibits\} 35% homologous to Caenorhabditis elegans gene unc\|89, is a novel specific antigen of A.caninum. Its ability to elicit the \{protective\} immunity and the probability of the recombinant protein as a vaccine need to be further evaluated.

Objective To build a bio-material artificial chest wall and discuss its feasibility and efficiency in reconstruction of huge bony defects of chest wall in mongrels in comparison with traditional "sandwich" procedure. Methods (1) The procine osteal and membranate tissues were treated with epoxy cross linking method and their surfaces were modified with amino acid solutions at various concen-trations and at different temperatures to obtain an artificial pleura and artificial ribs for construction of the artificial chest wall. (2) The huge bony defects (5 cm×5 cm) were created in chest wall of five Chinese mongrels. (3) Three mongrels in test group was repaired with artificial chest wall, while two mongrels in control group was repaired with traditional "sandwich" complex. A follow-up was carried out to observe reconstruction effect and rejection in both groups at 3,6 and 12 months after implantation. Results There was no death found during the perioperative period and at 12 month follow-up in test group, with abnormal contour of chest wall and good thoracic activity after reconstruction. In the meantime, there found no rejection, collapse in the repaired region or paradoxical respiration. The postoperative X-rays at 3,6,12 and 24 months showed a good integrity of the thorax, with no collapse, deformation or abnormal movement. Meanwhile, the follow-up of control group showed a normal contour but slight collapse, with no paradoxical respiration. The Chest X-ray examination revealed that the bone cement in" sandwich" complex was X ray opaque and showed mild abnormal movement with breathing. The common blood test and immune items showed no abnormal. Conclusions The bio-material artificial chest wall is a safe and effective reconstruction technique for bony defects of thoracic wall in mongrels, with no acute or chronic rejection.

Objective To assess the safety and efficacy of an olive oil-based lipid emulsion for parenteral nutrition in patients after esophagectomy.Methods In the randomized controlled trial,60 patients undergoing esophagectomy were divided into study group(n=30,received olive oil-based lipid emulsion)and control group [n=30,received medium-chain triglyceride/long-chain triglyceride(MCT/LCT)emulsion].The parenteral nutrition Was provided for 7-10 postoperative days.The nutritional formulas were equivalent in nitrogen,calorie,osmotic pressure,and fluid volume.Peripheral venous blood tests were performed before operation and on the first and eighth postoperative days.All the patients were evaluated by nutritional status(weight,body mass index,nutritional risk screening,etc.),safety profiles[full blood test,electrolytes,aspartate aminotransferase(AST),alanine amiotransferase(ALT),total bilirubin and direct bilirubin,blood urea nitrogen(BUN),creatinine,blood glucose,etc.],and efficacy indicators(hemoglobin,albumin,total protein,etc.).Results The albumin and total protein levels returned to the normal ranges in beth groups 8 days after operation,although both levels were significantly higher in study group(P=0.000).Also,the difference of total protein levels between the eighth and first postoperative days Was significantly higher in the study group(P=0.002).In addition,the AST and BUN readings returned to normal ranges 8 days after operation in the study group, which were significantly lower than those in control group (P = 0.025, P = 0.013).No serious adverse events were reported in both groups.Other nutritional parameters, renal and hepatic safety profiels, vital signs, and hematology showed no significant difference between two groups.Conclusions Olive oil-based lipid emulsion is a safe and efficient lipid emulsion for parenteral nutrition in patients undergoing esophagectomy.Compared with MCT/LCT, it has less effect on AST and BUN.

Objective To evaluate and screen out the specific characters of thyroid nodules on grayscale and color Doppler ultrasonography. Methods One hundred and four thyroid nodules in 95 patients were studied with gray-scale and color Doppler ultrasound before surgery.According to the probability of malignancy,the main independent characters were screened out by Logistic regression analysis and then got the formula which could calculate the probability of malignancy.Results Fifty three nodules,included 44 hyperplasia,7 adenomas,1 switch granuloma and 1 Hashimoto's disease,were benign.Fifty one nodules,included 47 papillary carcinomas,3 medullary carcinomas,1 metastatic carcinomas,were malignant pathologically.The significant characters to differentiate malignant from benign were the shape,margin,border,halo,echostucture,echogenecity,echo uniformity,calcicum,bolood vessel shape and partial abnormal rich blood of thyroid nodules.After two variables logistic regression analysis,a fomula were worked out.Probability of malignancy=1/1+e-z,which z=-7.44+1.30×halo+1.05×calcicum+1.5×vessel shape.If a thyroid nodule with no halo,microcalcicum and irregular vessel shape,the probability of malignancy was 0.99,while with complete halo,no calcicum and regular vessele shape,the probability of malignancy of the nodule was 0.02.Conclusions The halo,calcium and blood vessel shape of thyroid nodules were three independent characters of thyroid nodules to predict malignant thyroid nodules on gray scale and color Doppler ultrasound.

Objective:Mutations in epidermal growth factor receptor (EGFR) can predict tumor response to tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). However, not all cases of NSCLC with EGFR mutations can respond well;thus, discovering the heterogeneity of NSCLC at the molecular level is necessary. This study aimed to determine the discrepancy in EGFR mutations in primary tumors, N2 lymph nodes, and plasma samples. Methods:Primary tumors, N2 lymph nodes, and plasma samples obtained from 49 patients with stageⅢA-N2 NSCLC were analyzed for EGFR mutations in exons 19 and 21 by using mutant-enriched liquidchip technology. Results:In 49 patients, we detected 18 (36.7%) EGFR mutations in primary tumors, 11 (22.4%) mutations in N2 lymph nodes, and 2 (4.1%) mutations in plasma samples. Eleven (22.4%) cases showed discordance in EGFR mutations between primary tu-mors and N2 lymph nodes. In nine cases, EGFR mutations were detected only in primary tumors, whereas EGFR mutations were de-tected only in N2 lymph nodes in two cases. In addition, EGFR mutations were detected in the plasma samples of two patients, who al-so carry mutations in their primary tumors. Conclusion:A considerable proportion of NSCLC cases showed discrepancy in EGFR muta-tions between primary tumors and N2 lymph nodes. In addition, the detection rate of EGFR mutations was lower in plasma samples obtained from patients with stage IIIA-N2 NSCLC. All of the results indicated tumor heterogeneity at the molecular level during metas-tasis, and this heterogeneity may have implications during treatment with TKIs.

Objective: To explore the status of lymph node metastasis of middle third thoracic esophageal squamous cell carcinoma and its influence on the prognosis and to seek the reasonable range of lymphade-nectomy. Methods: A total of 129 patients who underwent curative esophagectomy with modern two-field lymphadenectomy of middle third thoracic esophageal squamous cell carcinoma were reviewed. Results: The lymph node metastasis rate was 56.6% and the upper mediastinal lymph node metastasis rate was 43.4%. The lymph node metastasis ratio (positive nodes/total dissected nodes, LMR) was 11.3%. Paraesophageal lymph nodes, lymph nodes near the right recurrent nerve, the left gastric and infracadnal lymph nodes were most commonly involved when the tumor was located in the middle thoracic esophagus. Tumor differentiation, the depth of tumor invasion and the length of tumor were influencing factors for lymph node metastasis. The 5-year survival of N_0, N_1 (LMR≤20%) and N_1 (LMR>20%) patients were 50.4%, 31.0% and 6.8%, respective-ly, with a significant difference among the three groups (P=0.000). Conclusion: LMR was one of the key fac-tors affecting the prognosis, of esophageal cancer. Patients with middle third thoracic esophageal carcinoma should be treated with radical surgery with modern two-field lymphadenectomy.

Objective:To study the adverse reactions (ADRs) of recombinant human erythropoietin (rHuEPO) to provide refer-ence for clinical rational and safe medication. Methods:ADRs induced by rHuEPO reported at home and abroad were collected and analyzed in respects of age,gender,original illness, occurrence time, clinical manifestations and the results. Results: After the re-trieval,there were 149 cases of rHuEPO-induced ADRs with the damage of cardio vascular system, hematologic system, skin and its appendents accounting for 43.4%,20.8% and 12.7%,respectively. The top three main clinical manifestations of rHuEPO drug reac-tions were hypertension,pure red-cell aplastic anemia (PRCA) and hyperkalemia. The occurrence time should be paid particular at-tention in 5-12 weeks after the administration (43.0%). Conclusion:Physicians should be aware of rHuEPO-induced ADRs (espe-cially the occurrence time),pay attention to patients' medication education and avoid serious adverse reactions.

Objective To explore the risk factors of the occurence and 28-day death of acute respiratory distress syndrome (ARDS) in intensive care unit (ICU). Methods A prospective multicentral cohort study was conducted. The patients from five ICUs of grade A tertiary hospitals in Beijing from July 2009 to March 2014, including sepsis,septic shock,trauma,pneumonia,aspiration,massive blood transfusion,bacteremia and pulmonary contusion,were enrolled. Researchers in each center reported the records with uniform tables,which included demographic,systemic conditions,the primary disease,and the severity within 24 hours,past history and so on. According to the admission diagnosis in ICU,these patients were divided into ARDS group and other severe disease control group. The risk factors of occurence and prognosis of ARDS were analyzed by univariate analysis,multivariate logistic regression and multivariate COX regression analysis. Kaplan-Meier method was applied to draw the 28-day survival curves of the two groups. Results There were 343 critical patients included in this prospective multicenter cohort study,of which 163 patients who developed ARDS were considered as ARDS group(2 case lost to follow-up, and 49 died)and 180 patients who did not developed ARDS regarded as severe control group(1 case lost to follow-up, and 34 died). The 28-day mortality of ARDS group was significantly higher than that of severe control group〔30.43%(49/161)vs. 18.99%(34/179),χ2=6.013,P=0.014〕. Multivariate logistic analysis showed that aspiration〔odds ratio(OR)=6.390,95% confidence interval(95%CI)=2.046-19.953,P=0.001〕,history of alcohol (OR=4.854,95%CI=1.730-13.617,P=0.003),sepsis(OR=2.859,95%CI=1.507-5.425,P=0.001), pneumonia(OR=2.822,95%CI=1.640-4.855,P<0.001),acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score(OR=1.050,95%CI=1.007-1.094,P=0.022)were significantly associated with increased risk of ARDS occurence. When respiratory rate>30 beats/min(OR=3.305,95%CI=1.910-5.721,P<0.001), heart rate>100 beats/min(OR=2.101,95%CI=1.048-4.213,P=0.037)happened in critically ill patients, it highly suggested ARDS would happen. The proportion of the patients whose serum creatinine>176.8 μmol/L in ARDS group was lower than that in control group(OR=0.387,95%CI=0.205-0.733,P=0.004). Multivariate COX regression analysis showed that old age and septic shock were significantly associated with the increased risk of in 28-day death of ARDS〔advanced age:hazard ratio(HR)=1.040,95%CI=1.018-1.064,P<0.001;septic shock:HR=3.209,95%CI=1.676-6.146,P<0.001〕. Kaplan-Meier showed that the survival patients in ARDS group was significantly lower than those in severe control group(χ2=7.032,P=0.008). Conclusions Among critical ill patients,aspiration,history of alcohol,sepsis,pneumonia,increased APACHEⅡ score were the risk factors of ARDS development. Respiratory rate>30 beats/min and heart rate>100 beats/min could predict the occurrence of ARDS in critical patients. Old age and septic shock were the risk factors of 28-day death of ARDS.

Objective To investigate the correlation between TYMS gene mRNA expression level and EGFR mutation in stage Ⅲ A-N2 non-small-cell lung cancer tissue (NSCLC).Methods The branched DNA-liquidchip technology (bDNA-LCT) and mutant-enriched liquidchip (MEL) technology were used to detect the TYMS mRNA expression and EGFR mutations at exon 19 and 21 in NSCLC tissues from 30 patients with stages Ⅲ A-N2 NSCLC,and the results were analyzed.Results Among 30 cases,low TYMS mRNA expression was in 14 cases (46.7 ％),middle to low TYMS mRNA expression in 7 cases (23.3 ％),middle mRNA expression in 7 cases (23.3 ％),middle to high TYMS mRNA expression in 0 case and high TYMS mRNA expression in 2 cases (6.7％);12 cases of EGFR mutation were detected out with the mutation rate of 40.0％,including 6 cases of exon 19 deletion and 6 cases of exon 21 missense mutation.The TYMS mRNA expression level was correlated with the EGFR mutation.The EGFR mutation commonly occurred in the tumor tissue of the patients with TYMS mRNA low expression (Z=-2.604,P=0.009).Conclusion The TYMS mRNA expression in NSCLC tissue is correlated with the EGFR gene mutation,which can provide references for the drug selection aiming at the patients with different conditions.

BACKGROUNDMost stage IIIB (T4/N3) non-small cell lung cancer (NSCLC) can not be cured via operation, but how is the outcome for those with T4 or occasionally N3 which can be completely resected? This paper retrospectively analyses the effects of the tumor characteristics and postoperative treatments on the survival of 51 patients with stage IIIB NSCLC completely resected in this hospital from January 1, 1997 to April 30, 2001.

METHODSThe effects of clinical pathophysiological characteristics such as gender, histological type, differentiation, T-stage and lymph node status and the postoperative chemotherapy or radiotherapy on the prognosis of 51 patients with completely resected stage IIIB NSCLC were retrospectively analysed.

RESULTSThere were no statistic survival differences in the disease characteristics such as the different gender (Log rank=0.992, P=0.319), histological types (Log rank=1.263, P=0.260), differentiation (Log rank=1.104, P=0.577), T-stage (Log rank=0.106, P=0.588) and lymph node status (Log rank=1.297, P=0.731), also no difference between groups whether or not there was postoperative mediastinal radiotherapy (Log rank=0.482, P=0.488) or postoperative chemotherapy (Log rank=0.051, P=0.759).

CONCLUSIONSNeither the tumor characteristics such as gender, histological type, differentiation, T-stage and N-stage, nor the postoperative mediastinal radiotherapy or chemotherapy affect the survival of stage IIIB NSCLC with complete resection.