0.05). Compared with the sham operative group, NO content in model group was obviously lower( P

In modern biology and biotechnology research, recombinant gene expression has been the most popular method to obtain the target protein. In recent years, many foreign genes have been efficiently expressed in Escherichia coli. However, proteins encoded by animal, plant or mesophilic microbial genes often lose activities or become denatured within a few hours at regular growth temperatures for E. coli; some other target proteins are toxic to host cells and therefore difficult to be over-expressed. The new T-vector, pEXC-T, was constructed by combining TA cloning and cold-shock induction to obtain high expression levels with low costs. This paper reports the construction of pEXC-T and optimization of induction techniques for gene expression. Two instable proteins were tested and successfully expressed in soluble form by using pEXC vector. The development of pEXC-T offers a convenient technique for the preparations of recombinant proteins to be used in structure/function studies, or as diagnostic markers and medicinal proteins.
Biotechnology ; Cold Temperature ; Escherichia coli ; genetics ; Gene Expression ; Genetic Vectors ; Plasmids ; genetics ; Recombinant Proteins

Objective To investigate the role of acid and bile reflux in children with gastroesophageal reflux disease (GERD) and to evaluate the significance of detecting acid and bile reflux in diagnosing GERD in children.Methods Using ambulatory 24 h pH mo-(nitoring) and bilirubin monitoring technique, we simultaneously assessed the changes of intraesophageal pH and bile reflux in 23 subjects (including 11 healthy controls and 12 patients with GERD).Results The time of esophageal acid exposure (pH

Child, Preschool ; Female ; Humans ; Sarcoma ; diagnosis ; surgery ; Stomach Neoplasms ; diagnosis ; surgery ; Stromal Cells ; pathology

Objective To evaluate the additional value of CACS in detection of coronary artery disease (CAD) with MPI.Methods A total of 188 suspected CAD patients (128 males,60 females;average age (61.93±9.16) years) who underwent one-step examination of MPI combined with CACS from December 2012 to August 2014 were enrolled in this retrospective study.According to the gold standard of CAG,the diagnostic efficacy of MPI was calculated.ROC analysis was performed to determine the optimal CACS threshold for the detection of CAD.Mann-Whitney u test and x2 test were used for statistical analysis.Results (1) Seventy-three CAD cases were confirmed (≥ 50％ stenosis) among 188 patients.The sensitivity,specificity,accuracy for CAD diagnosis with MPI were 65.8％ (48/73),75.7％ (87/115),71.8％ (135/188),respectively.Twenty-five CAD patients had negative findings with MPI,including 2 with LM disease,4 with three-vessel disease (LAD±LCX±RCA,3-VD),3 with 2-VD,16 cases with 1-VD.Among them 13 cases (52.0％,13/25) had intermediate lesions of 1-VD (50％ ≤ stenosis＜70％).(2) The CACS of CAD group was significantly higher than that of non-CAD group (172.40(19.25,516.45) vs 0;z=-8.465,P＜0.001).According to the ROC analysis,95.1 was the optimal CACS cutoff to detect CAD patients.Combining MPI with CACS (at cutoff of 95.1) improved the sensitivity of MPI (80.8％,59/73;x2 =4.233,P＜0.05) for the detection of CAD,with no significant decrease in specificity and accuracy (71.3％,82/115;75.0％,141/188;x2 values:0.558 and 0.490,both P＞0.05).(3) Of the 25 CAD patients with negative MPI results,11(44.0％,11/25) showed abnormal CACS(CACS≥95.1),consisted of 2 cases of LM disease,4 cases of 3-VD,2 cases of 2-VD,3 cases of 1-VD.Diagnosis was corrected by CACS in 8/9 cases of severe CAD (LM CAD or multivessel disease) which were missed by MPI.Conclusion CACS could offer additional information for MPI in detection of suspected CAD patients,which can improve the sensitivity of MPI for diagnosing CAD,especially for severe CAD with LM lesions or multivessel CAD.

Abdominal Pain ; etiology ; Child ; Child, Preschool ; Endoscopy, Gastrointestinal ; Female ; Humans ; Male ; Purpura, Schoenlein-Henoch ; complications ; pathology

OBJECTIVETo investigate the mRNA expressions of RASSF1A, Galectin-3 and TPO in papillary thyroid carcinoma and some other thyroid benign lesions, and evaluate their diagnostic significance.

METHODSReverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of RASSF1A, galectin-3 and TPO in the samples from 73 cases, including 23 cases with papillary thyroid cancer, 16 with nodular goiter, 29 with thyroid adenoma and 5 with Hashimoto's disease.

RESULTSA statistically significant difference in the mRNA expression of RASSF1A, Galectin-3 and TPO was observed between papillary thyroid carcinoma and follicular benign lesions (P<0.05). However, there was no significant difference among various kinds of benign lesions (P>0.05). A negative correlation of the expression of RASSF1A and Galectin-3 mRNA was found between thyroid benign lesions and malignant ones (P = 0.000). While the mRNA expression of RASSF1A and TPO was positively correlated between benign and malignant lesions (P = 0.028).

CONCLUSIONLoss of expression of RASSF1A and TPO mRNA but high expression of Galectin-3 mRNA in papillary thyroid carcinoma are common. Therefore, the products of these three genes may be closely related to the development of thyroid papillary carcinoma, and may be used as useful markers in differential diagnosis of papillary thyroid carcinoma from the benign lesions. The results are more reliable if this detection method is used in combination with other techniques.

Adolescent ; Adult ; Aged ; Autoantigens ; genetics ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Galectin 3 ; genetics ; metabolism ; Goiter, Nodular ; genetics ; metabolism ; pathology ; Hashimoto Disease ; genetics ; metabolism ; pathology ; Humans ; Iodide Peroxidase ; genetics ; metabolism ; Iron-Binding Proteins ; genetics ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; Tumor Suppressor Proteins ; genetics ; metabolism ; Young Adult