Objective:To obtain antibodies against amylin from a 'naive' human Fab fragment antibody phage diasplay library and to analyze the specificity of antigen binding activity.Methods:Panning and screening Fab antibody from the antibody library,the positive clones with well reactivity to amylin were selected after five times selection of 'adsorption-elution-enrichment'.Then the plasmid DNA which was extracted from the clones,was digested with Spe Ⅰ and Nhe Ⅰ to delete gⅢ (about 660 bp).The digested 47 000 bp DNA which was purified after separation of bands from agarose gel was ligated with T4-DNA ligase.The constructed expressing phagemids were transformed to the BL21(DE3)pLysS,soluble Fab was expressed in it by the induction of IPTG and its characteristics and specificity were determined by ELISA and Western blot.Results:Soluble Fab antibodies were expressed in E.coli.According with molecular weight of IgG Fab,protein band of about 47 kD was shown by SDS-PAGE.Western blot using the goat anti human IgG-HRP showed their binding activities.ELISA showed their specificity with amylin antigens and they did not react with bovine serum albumin.Conclusion:The high level expression and identification of the soluble human anti- amylin Fab fragment antibodies has been obtained successfully,which lays a solid foundation for further researching about the biological and pathological activities of amylin.

Objective To quantitatively analyze the image quality of megavoltage cone-beam CT (MVCBCT) under different scanning conditions to provide reference in clinical applications. Methods Si-emens ONCOR linear accelerator with MVCBCT was used to scan the phantom under different conditions. The image quality was evaluated in terms of image noise, uniformity, spatial resolution, contrast resolution, the number of Monitor Units(MUs) used in imaging,and the size of the reconstruction matrix. The comparison of the image quality between MVCBCT and conventional simulator CT was also analyzed. Results The image noise was decreased with the increase of the number of MUs. The uniformity index showed that the system u-niformity was weakly dependent on MU numbers or the size of the reconstruction matrix. Except for the ima-ges with 5 MUs,all other images had the spatial resolution of 0.4 lp/mm with a reconstruction matrix of 256 ×256. Better low contrast resolution was achieved by using more MUs. For typical pelvis and head-and-neck patients,the imaging dose at the center was 0.8 cGy/MU and 0.7 cGy/MU, respectively,and the maxi-mum dose was about 1.2 cGy/MU. For typical abdomen patients,the image maximum dose and center dose was 1.3 cGy/MU and 0.7 cGy/MU,respectively. Conclusions The image quality of MVCBCT is inferior to the conventional kilo-voltage CT. However,with the optimization of the parameters in imaging,we can a-chieve sufficient image contrast in the bone,air and some soft-tissue structures with low imaging dose to pa-tients. Such images can be used for IGRT.

OBJECTIVETo assess the efficacy and safety of Xiaozheng pills in treating mastoplasia.

METHODClinical trials were carried out by five hospitals. In each hospital, patients were divided into two groups with one group 24 patients (trial group) and the other 24 patients (control group). Total 240 patients were included in the study. According to randomized, double-blinded and placebo-controlled clinical study, the trial groups were treated by Xiaozheng pills with Rujiekang mimetic (placebo) and the control groups were treated by Rujiekang with Xiaozheng pills mimetic (placebo). Symptoms, laboratory test results as well as ADR were evaluated after 1 period of treatment.

RESULTThe overall response rates of trial group and control group were 93.8% and 88.6% respectively, no statistic difference between the two groups. No deleterious effect in both groups and the indexes of safety were normal.

CONCLUSIONThese results suggest that Xiaozheng pills are effective and safe in treating mastoplasia caused by qi stagnation, blood stasis or/and stagnation of phlegm.

Adolescent ; Adult ; Breast ; pathology ; Double-Blind Method ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Female ; Fibrocystic Breast Disease ; drug therapy ; pathology ; Humans ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the incidence of lymph node metastasis (LNM) in the right para-tracheal triangle (RPT) of esophageal carcinoma patients and the technique of dissection.

METHODSOn the top of double mediastinal and abdominal lymphadenectomy, 333 esophageal carcinoma patients received RPT lymphadenectomy through the right pleural apical approach from 1990 to 2001.

RESULTSIn these 333 patients, the lymph node metastasis (LNM) rate in the RPT was 36.40%. A total of 457 nodes among 2 159 nodes removed gave a metastasis degree of 24.96%. The LNM rates in RPT for cervical, upper third, middle third, and lower third segments of esophagus were 66.67%, 45.45%, 34.19% and 15.79% (P < 0.05), while their respective metastasis degrees were 44.44%, 27.04%, 24.32% and 18.92% (P > 0.05). The frequency of positive nodes in the RPT for PTI, PT1, PT2, PT3 and PT4 was 0, 17.24%, 28.7%, 45.16% and 53.57%, while those of metastasis degree were 0, 8.77%, 17.62%, 33% and 41.17% (P < 0.01). The frequency of LNM in the RPT in papillary, erosive, patch-like and covert type of early tumor was 40%, 3.85%, 0 and 0 (P < 0.05), while those of the metastasis degree were 29.41%, 1.82%, 0 and 0 (P < 0.01). Higher rate of LNM in progressive stenotic esophageal carcinoma was observed compared with those of the other gross types (56.52%, P < 0.05), so was the degree (P < 0.01). The frequency of LNM in the RPT for mono-focal and multi-focal tumor was 34.98% and 70% without significant difference (P > 0.05), while the degree was 24.29% and 53.33% (P < 0.05). Postoperative complications were: leak (0.6%), and recurrent laryngeal nerve injury (1.2%). No injury of vein or infra-clavicular artery, tracheal damage or mortality occurred.

CONCLUSION1. The lymph node metastasis from esophageal carcinoma has a tendency of wide spread and right para-tracheal triangle is an important region to be doomed. 2. With location, depth of tumor invasion and differentiation of tumor as major factors affecting LNM of esophageal carcinoma, dissection of this region should be paid more emphasis. 3. In early lesions, higher frequency of LNM in the RPT is found in papillary and erosive lesions than in the other macroscopic types. 4. Exposing the RPT, lymph node by dissection through a right pleural apical approach is very important and significant.

Adult ; Aged ; Cardia ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; methods ; Esophagus ; pathology ; Female ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Mediastinum ; Middle Aged ; Neck ; Neoplasm Invasiveness

The prevalence of thyroid cancer has shown an upward trend in China in recent years. Advances in thyroid ultrasound and fine needle puncture cytology have improved the accuracy of the preoperative diagnosis of thyroid cancer. Also,the application of endoscopy-assisted techniques and intraoperative nerve monitoring technology and the further understanding of thyroid lymph node metastasis have made the thyroid surgeries safer and less invasive. This article summarizes the recent advances in the surgical therapy of thyroid cancer.
Carcinoma, Papillary ; surgery ; Humans ; Iodine Radioisotopes ; therapeutic use ; Molecular Targeted Therapy ; Neoadjuvant Therapy ; Thyroid Neoplasms ; surgery ; therapy

Objective To evaluate the feasibility and diagnostic accuracy of CE-MRA with low dose contrast agent by comparison with DSA in diabetic patients with peripheral arterial diseases. Methods ( 1 )Study in vitro: test tubes containing Gd-DTPA of different concentrations were scanned, and the relationship between signal intensities and concentrations of GD-DTPA was analyzed. DSA and CE-MRA with selected concentrations of Gd-DTPA were performed on stenotic vascular models to estimate the proper low dose of GD-DTPA for clinical applications. (2) Clinical applications: 78 diabetic patients with peripheral arterial diseases were scanned from the abdomen and pelvis station to the calf-foot station in a 3 T MR system with standard bolus chase 3D CE-MRA sequence after injection of 13 ml GD-DTPA . The image quality,diagnostic rate of stenosis of arteries in calf and degree of venous contamination were evaluated with Fisher's exact test. DSA images of 220 vascular segments in 22 patients ( 10 segments per patient) were acquired as the gold standard and compared with CE-MRA by using Kappa test. Results The MR signal intensities were proportional to the concentrations of contrast agent in present study, and all stenotic segments of vascular model were displayed by CE-MRA with GD-DTPA at lower concentration of 1.5 mmol/L. As for MRA images of 78 diabetic patients with low dose Gd-DTPA, about 97.4% (76/78) showed diagnostic image quality for pelvic and thigh stations. But the MRA images of lower extremities were interfered by the venous contamination significantly (P ＜ 0.01 ). Compared with DSA for 22 patients, the diagnostic sensitivity, specificity and agreement coefficient (Kappa value) of MRA were 96. 0% ( 168/175), 73.3%(33/45), and 0.72 (P＜0.01), respectively. Conclusion Using 3.0 T MR scanner, high quality CE-MRA of lower limb arteries can be obtained for clinical applications with contrast agent dose as low as 13 ml,which has comparable diagnostic sensitivity and specificity with DSA. But the limitation of venous contamination in MRA image should be resolved in further studies.

Objective: To understand the parenting styles of junior high school students in Beijing, to explore the relationship between parenting styles with students academic performance and the role of mental health in this association. Methods: From October 2019 to January 2020, a stratified random cluster sampling method was used to select 5 196 students in the first and second grades of 11 public junior high schools in urban and rural areas in Beijing, and Slort Egna Minnenar Barndoms Uppfostran-China,Middle School Students Mental Health Scale-60 and self designed questions were used in the questionnaire survey. Results: The average score of parenting style rejection dimension was (9.44±3.88), and the average score of emotional warmth dimension was (19.85±5.66). The average score of comprehensive score of students mental health was ( 1.92 ±0.73). Mental health score of girls was higher than boys (1.97±0.74)(1.87±0.71), and the differences were of statistical significance( t=5.06, P <0.01). Parental rejection and emotionally warm parenting styles were significantly correlated with students academic performance （ β =-0.54，1.15， P <0.01). Mental health played a negative moderating role between parenting style and students academic performance. The better students mental health were, the greater the influence of parenting style on academic performance. Conclusion Parenting style is related to children s academic performance. Parents should give their children more affirmation, encouragement and praise, in order to improve their academic performance.

OBJECTIVETo investigate anticancer effect and molecular mechanism of N-[(Cyclohexyloxy)-4-nitrophenyl] methanesulfonamide on HepG2 cells in vitro.

METHODSHepG2 cells were treated with various concentrations (100, 200, 300, 400 micromol/L) of NS-398 [selective for cyclooxygenase 2 (COX-2) inhibition]. Cell growth was measured by MTT method, DNA fragmentation gel analysis was used to analyze the apoptosis cells, DNA ploidy and apoptotic cell percentage were examined by flow cytometry (FCM). PGE2 concentration was measured by radioimmunoassay method. The expressions of COX-2 were also examined by Western blot analysis.

RESULTSNS-398 inhibited HepG2 cell proliferation and induced apoptosis in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased and quiescent G1 phase was accumulated with NS-398 concentration increasing. The IC50 of 24 hours was 300 micromol/L. The release of PGE2 was significantly reduced in HepG2 cells with the values of NS-398 being (0.70 +/- 0.02), (0.48 +/- 0.02), (0.29 +/- 0.01) and (0.18 +/- 0.01) respectively, as compared with control group (0.03 +/- 0.01). NS-398 could inhibit the activity and expression of COX-2, with higher concentration, it can significantly down-regulate the expression of COX-2 (t = 3.736, 1.623, 1.810, 2.587, P < 0.01).

CONCLUSIONNS-398 might significantly inhibit the proliferation of HepG2 cells and induce apoptosis. The mechanisms were related with the accumulation of quiescent G1 phase and the inhibition of COX-2 activity.

Apoptosis ; drug effects ; Cell Line, Tumor ; metabolism ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; biosynthesis ; Humans ; Nitrobenzenes ; pharmacology ; Sulfonamides ; pharmacology

OBJECTIVETo summarize the surgical effect and clinical application value of esophagectomy with extended 2-field lymph node dissection for patients with esophageal carcinoma.

METHODSFrom June 1987 to December 2008, 1690 patients with esophageal cancer underwent esophagectomy with extended 2-field (thoracic and abdominal) dissection of lymph nodes. Patients with the middle and lower thoracic esophageal cancer underwent Ivor-Lewis esophagectomy, and patients with upper thoracic esophageal cancer underwent Akiyama esophagectomy. 2-field (thoracic and abdominal) lymph node metastases information and the 1, 3, 5, 10-year survival rates were analyzed retrospectively.

RESULTSLymph node metastases were found in 713 patients. The lymph node metastases rate was 42.2% (713/1690).Thoracic lymph node metastasis rate was 39.3% (665/1690), among which in the right pleural apical para-tracheal triangle was 20.7% (349/1690), in the posterior upper mediastinum was 26.3% (444/1690), in the lower mediastinum was 18.2% (307/1690). Abdominal lymph node metastasis rate was 20.1% (339/1690). THE Postoperative complication rate was 16.4% (278/1690), among which the pulmonary complication rate ranking the first, was 43.6% (136/312). The operative mortality rate was 0.2%. The 1-year, 3-year, 5-year and 10-year survival rates were 88.2% (1388/1574), 63.5% (868/1367), 54.8% (705/1287) and 30.8% (232/754), respectively. The 5-year survival rate in patients without lymph node metastasis was 76.2% (448/588), but that in patients with lymph node metastases was 36.8% (257/669).

CONCLUSIONThe results of this study demonstrated that Ivor-Lewis and Akiyama esophagectomy with two-field lymph node dissection exposes the operation fields clearly and make radical lymphadenectomy thoroughly, especially the lymph nodes in the posterior upper mediastinum around the recurrent laryngeal nerve and in the right pleural apical para-tracheal triangle. It is essential that patients with esophageal carcinoma with lymph node metastases should undergo esophagectomy with extended 2-field dissection of lymph nodes. This can elevate the postoperative 5-year survival rate remarkably.

Adenocarcinoma ; mortality ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; mortality ; pathology ; surgery ; Esophageal Neoplasms ; mortality ; pathology ; surgery ; Esophagectomy ; adverse effects ; methods ; Female ; Humans ; Lymph Node Excision ; adverse effects ; methods ; Lymphatic Metastasis ; Male ; Middle Aged ; Respiratory Insufficiency ; etiology ; Retrospective Studies ; Survival Rate

CpTI (Cowpea Trypsin Inhibitor) is a widely used insect resistance gene in the plant genetic engineering for its high insecticidal activity and the minimal ability of the insects to evolve resistance to it. To facilitate the safety assessment of genetically modified foods (GMFs) with CpTI protein, we need to produce gram quantities of this protein in microbes. With the pGEX fusion expression system, we expressed the GST-CpTI protein in E. coli BL21, which accounted for approximately 40% of germ proteins. By Glutathione Sephrose 4B affinity chromatography, GST-CpTI was obtained with the purity up to 90%. Overnight incubate the fusion proteins with Thrombin protease, we got the CpTI proteins cleavage of GST tag. Both of the GST-CpTI and CpTI proteins showed notable trypsin inhibitor activity. Immunization of rabbits with purified fusion protein generated high titer antibodies (> 20000), measuring by ELISA. Western Blotting also showed specific Ag-Ab binding band between the antiserum and the CpTI proteins no matter in the whole supersonic germ proteins or purified from the column. All these made a good ground for the further safety assessment of CpTI protein.
Animals ; Blotting, Western ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Trypsin Inhibitors ; genetics ; metabolism