Evaluation of biological activity of an eukaryotic expressed anti-H5N1-M1 cell entry single molecule antibody / 中国生物制品学杂志

@#Objective To evaluate the biological activity of a eukaryotic expressed anti-H5N1-M1 cell entry single molecule antibody(TAT-ScFv-mFc). Methods The immune binding activity and affinity of TAT-ScFv-mFc to H5N1-M1 protein were detected by Western blot and localized surface plasmon resonance(LSPR)respectively;The inhibitory effect of TAT-ScFv-mFc on influenza virus H1N1 was detected by CCK-8 assay;The membrane penetration ability of TAT-ScFv-mFc to MDCK cells was verified by immunofluorescence assay. A total of 30 female BALB/c mice were injected with TAT-ScFv-mFc via tail vein,200 μL per mouse. Blood samples were collected at 5,60,120,240 and 360 min after injection. Serum samples were separated and detected for the titers by ELISA,and the half-life of TAT-ScFv-mFc was calculated according to the half-life curve drawn by Origin 2021 software. Results TAT-ScFv-mFc showed specific binding to H5N1-M1 protein with a binding rate constant of 6. 67 × 10~4[1/（M*s）]. The survival rate of MDCK cells infected by H1N1 increased gradually with the increase of TAT-ScFv-mFc concentration in a dose-dependent manner,which obviously inhibited the replication of H1N1. TAT-ScFv-mFc penetrated the cell membrane of MDCK cells in a short time,entered the cell and bound to virus M1protein,thus inhibiting virus replication and assembly. The half-life of TAT-ScFv-mFc in mice was 212 min. Conclusion TAT-ScFv-mFc has good immune binding activity and affinity with H5N1-M1,can effectively inhibit the replication of H1N1,has good penetration ability to MDCK cell membrane,and has a long half-life in mice,which lays a foundation of the drug treatment,vaccine research and preventive treatment of H5N1 infection.

Effect of modified NDV F48E9 strain HN gene and in vitro expression of its DNA vaccine / 生物工程学报

Improving expression of antigen is critical to the immunogenicity of DNA vaccines. To achieve this goal, we modified the NDV F48E9 strain HN gene by optimizing the condon usage and inserting the secretary leader sequence [A/Goose/Guangdong/1/96 (H5N1) HA gene, Accession No. AF144305]. The HN gene modified and knocked the signal peptide off were named SoptiHN and optiHN. The three sequence: SoptiHN, optiHN and the NDV F48E9 strain HN gene were inserted into the vector pVAX1 and vector pVAX1-CpG including CpG-ODN sequence respectively. Then we got six recombinant plasmids: pV-SoptiHN, pVC-SoptiHN, pV-optiHN, pVC-optiHN, pV-HN and pVC-HN. By optimizing condon usage in transiently transfected 293T cells, expression levels of HN gene were higher from the codon-optimized gene than the counterpart. Moreover, both optimization of condon usage and addition of signal peptide could improve expression of HN gene in vitro.

Establishment of Cisplatin Resistant Head and Neck Cancer Cell Lines and Cross-resistance of Docetaxel / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Treatment of head and neck cancer with cisplatin-based combination chemotherapy has yielded increased complete response rates. In spite of the initial good response, recurrent cancer after chemotherapy does not respond to cisplatin based chemotherapy. In recent years, the analogues of taxol have used in a new chemotherapy treatments for head and neck cancer. The purpose of this study was to develop the acquired resistance to cisplatin in head and neck cancer cell line and to study the cross-resistance of docetaxel and 5-fluorouracil (5-FU) as compared to the parental cells in vitro. SUBJECTS AND METHOD: Two different human head and neck cancer cell lines (PNUH-12, SNU-899) were studied. We developed two cisplatin resistant cell lines (PNUH-12 R1, SNU-899 R1) by exposing them to near continuous increasing concentration of cisplatin. The morphology, growth curve of resistant cell lines were studied and the resistance of these cell lines to the cisplatin, docetaxel and 5-FU were compared to that of parental cells by MTT assay. RESULTS: The pattern of growth and shape of PNUH-12 R1 and SNU 899 R1 cell lines showed similarity to their parent cell lines. The growth curve of these cell lines were suppressed when compared with the parental cells. When compared to the parental cells, PNUH-12 R1 cells were 2.0-fold, 3.8-fold, and 1.4-fold resistant, and SNU 899 R1 cells 3.5-fold, 6.7-fold, and 15.0-fold resistant to cisplatin, docetaxel and 5-FU, respectively. CONCLUSION: In our study, cisplatin resistant cell lines of head and neck cancer have strong cross-resistance to docetaxel. However, there was difference in cross-resistance to 5-FU between these cell lines.

Clinical Significance of p53 Gene and nm23 Gene Expression in Esophageal Cancer / 대한흉부외과학회지

BACKGROUND: Although significant progress has been made in the surgical treatment of esophageal carcinoma as well as in the detection of early stage esophageal carcinoma by diagnostic techniques, the prognosis of the esophageal carcinoma patients remain poor. The p53 gene product is known to regulate cell growth and proliferation. And the nm23 gene was identified originally as an anti-metastatic influence whose expression was correlated inversely with tumor metastatic potential in murine melanoma cell lines. This experiment was intended to know the relationship among the p53 and nm23 gene expression versus clinicopahologic characteristics of the esophageal cancer. MATERIAL AND METHOD: Total 40 cases were collected from patients who had undergone esophagectomy at St. Mary's Hospital, Catholic university of Korea. Immunohistochemical stain for p53 mutant-type protein and nm23 proein was graded as 50% positive tumor cells: +++ . The tumor invasion was grades as none: - ; mild: + ; moderate: ++ ; severe: +++ . RESULT: Overexpression of p53 protein and nm23 was not associated with the survival and cliniocopathologic characteristics of the esophageal cancer. Moreover, the combination analysis of p53 and nm23 revealed that there was no relationship between the gene expression and the clinicopatholic characteristics of the esophageal cancer.

Delayed Primary Repair of Perforated Epiphrenic Diverticulum

A 68-yr-old man complaining of sudden, postprandial chest pain visited the emergency room. His symptom had been aggravated during the preceding two days. Upper gastrointestinal contrast study with gastrographin showed leakage of dye from the epiphrenic diverticulum in the lower third of the esophagus. The primary repair was urgently carried out. Upper gastrointestinal contrast study 14 days after operation revealed an esophageal leakage which was small and confined. The patient was managed with conservative treatments such as intravenous hyperali-mentation and broad-spectrum antibiotics. Forty-two days after the operation, a gastrographin swallow study showed the absence of leaks. This is the first report-ed case of a perforated epiphrenic esophageal diverticulum repaired by delayed primary repair in Korea.

Prognosis Factors of Tricuspid Regurgitation after the Operation for Left-sided Valvular Heart Disease / 대한흉부외과학회지

BACKGROUND: Tricuspid regurgitation has been considered as a secondary lesion when it is combined with left valvular heart diseases. However, there have been some reports which show that tricuspid regurgitation keeps going and results in congestive heart failure even after a successful operation for left valvular heart disease. So far, there are no definite operation indications and predictive factors for the tricuspid regurgitation which is resulted from the left sided valvular heart disease. We designed this study to evaluate the effects of pulmonary artery pressure and left ventricular ejection fraction on the prognosis of tricuspid regurgitation, and to make an operation indication for the patients with secondary tricuspid regurgitation. MATERIAL AND METHOD: We reviewed the medical records of patients who underwent surgery for the left sided valvular heart disease with tricuspid regurgitation and were followed for more than 1 year with echocardiograms. There was a total of 114 cases. We compared the grades of tricuspid regurgitations and pulmonary artery pressures and left ventricular ejection fractions on the basis of echocardiograms which were checked preoperatively and on the last follow up. RESULT: There were 43 cases of tricuspid annuloplasty. In these patients, the grades of tricuspid regurgitations were improved in 42 cases (97.7%). But in 71 cases without annuloplasty, 29 cases (41%) were improved, 32 cases (45%) had no change, and 10 cases (14%) were aggravated. This finding shows significant differences in the prognoses of tricuspid regurgitations between the two groups (p0.05). The improvements of tricuspid regurgitations are not statistically related to the changes of pulmonary artery pressures or left ventricular ejection fractions. CONCLUSION: This study shows that it is impossible to predict the prognoses of tricuspid regurgitations with preoperative pulmonary artery pressures or left ventricular ejection fractions. Also, the excellent results of tricuspid annuloplasty is proven in controlling the secondary tricuspid regurgitations. Therefore, when tricuspid regurgitation is detected preoperatively, the procedures to correct the tricuspid regurgitation at the time of the operation for the left-sided valvular heart disease must be considered positively, regardless of the grades of tricuspid regurgitations, to prevent significant tricuspid regurgitation that may develop later.

A Study on Dietary Isoflavone Intake from Soy Food and Urinary Isoflavone Excretion and, Menopausal Symptoms in Korean Women in Rural Areas / 대한지역사회영양학회지

Very little is known about the relation between isoflavone intake and menopausal symptoms in Korean woman To find the effects and correlations between these factors, questionnaires(maternal factors, menopausal symptoms) anthropometric measurement, 24hr dietary recalls, and urinary isoflavones analysis were conducted in 100 postmenopausal women residing in the Kyunggi-do rural area. The average age hight and weight of the subject were 61.5 years, 153.6 cm and 56.2kg. The average age at menarche, menopause, and menstrual cycle were 16.7 years, 47.2 years, 29.8 years, respectively. Most of the postmenopausal women experienced mildly menopausal symptoms. The mean calorie, protein and calcium intake were 1417.6 kcal(74.7% of the Korean RDA) 53.2 g(88.6% of the Korean RDA) and 454.0mg(65.6% of the Korean RDA) respectively. The ration of energy from carbohydrate, protein, and fat was 68: 15: 17 The average isoflavone intake from soy foods was 27.27 mg. The major food source of the isoflavone were soybeans and soybean curd. The average urinary isoflavone excretion was 2. 78nmol/mg Cr and showed significant positive correlation with isoflavone intake. Isoflavone intake from soy foods had a negative correlation with the severity of menopausal symptoms. Therefore, soy foods which contain isoflavone may have a protective effect on menopausal symptoms of women in Korea.