Molecular response of human cervical and lumbar nucleus pulposus cells from degenerated discs following cytokine treatment

We investigated the molecular response of degenerated human cervical and lumbar nucleus pulposus (NP) cells following cytokine treatment. Degenerated cervical and lumbar discs (8 each) were obtained from patients who underwent discectomy for degenerative disc disease; NP cells were isolated and cultured. The mRNA expressions of aggrecan, alkaline phosphatase, type I collagen, type II collagen, osteocalcin, and Sox9 in the 2 groups were compared by real-time PCR, before and following treatment with rhBMP-2 and TGF-β1. Immunoreactivity was analyzed to check protein activity. Type I collagen expression was significantly higher in cervical compared with that in lumbar disc cells. The mRNA expression was significantly increased after rhBMP-2 and TGF-β1 treatment. After rhBMP-2 treatment, mRNA expression of type I and II collagens increased significantly more in cervical than in lumbar NP cells. Following TGF-β1 treatment, the increase in mRNA expression was not significantly different between cervical and lumbar disc cells. Protein immunoreactivity, before and after cytokine treatment was similar to mRNA expression data. The matrix-related gene expression of cervical and lumbar NP after rhBMP-2 and TGF-β1 treatment increased similarly, with the exception of collagen expression.ope

Association between BMP-2 and COL6A1 gene polymorphisms with susceptibility to ossification of the posterior longitudinal ligament of the cervical spine in Korean patients and family members.

COL6A1 and BMP-2 genes have been implicated in ossification of the posterior longitudinal ligament (OPLL) susceptibility in Japanese and Chinese Han populations. However, no study has yet investigated the DNA of unaffected family members of patients with OPLL. This study investigated differences in genetic polymorphisms of BMP-2 and COL6A1 between Korean patients with OPLL and their family members (with and without OPLL). A total of 321 subjects (110 patients with OPLL and 211 family members) were enrolled in the study. Associations between two single nucleotide polymorphisms (SNPs) of the BMP-2 gene (Ser37Ala and Ser87Ser) and two SNPs of COL6A1 [promoter (-572) and intron 33 (+20)] with susceptibility to OPLL of the cervical spine were investigated between the two groups (OPLL+ and OPLL-). Of the 321 subjects, 162 had cervical OPLL (50.4%; 110 patients, 52 family members). There was a familial tendency of OPLL in 34 of the 110 families (30.9%). Allele and haplotype frequencies of the four SNPs in the BMP-2 and COL6A1 genes did not differ significantly between the OPLL+ and OPLL- groups, even when excluding participants over 50 years of age. This is the first report identifying SNPs of COL6A1 and BMP-2 in Korean patients and family members with OPLL. Although allele and haplotype frequencies were similar with those of a previous study in Japanese and Chinese patients, unaffected family members also showed similar rates of these SNPs in the present study. These results suggest that these SNPs may not directly influence the expression of OPLL.ope

한국의 평생교육 정책 발전 과정 및 특성 분석

학위논문(석사)--아주대학교 교육대학원 :평생교육전공,2001Maste

일본형 고용시스템하에서의 고연령자 고용정책

일본형 고용시스템은 입구(신규졸업자 일괄 채용)부터 출구(정년퇴직)까지, 그리고 그사이의 처우 (연공임금, 연공서열)까지도 정년제도를 기축으로 하여 연령 규범에 근거한 독자적인 법 정책을 완성해 왔다. 이러한 상황 속에서 최근 저출산・고령화가 심각한 사회문제로 지적됨에 따라, 종래와 같은 연령 규범에 근거한 정책으로는 저출산・고령화로 인한 노동력 부족 문제 등을 해결하기 어려울 뿐만아니라 최근의 연금 정책의 전개와 맞물려 고용에서의 연령 제한을 시정해야 할 필요성과 함께, 애초에 연령 규범에 근거한 정책은 연령차별에 해당할 수 있다는 지적이 꾸준히 제기되어 왔다. 이에 본 논문에서는 연령 규범을 중시하는 일본형 고용시스템하에서 관련 정책 및 법률이 어떻게 전개되어 왔는지를 살펴본 후, 이를 둘러싼 판례의 동향 및 학설의 논의 상황에 대해서 검토하였다. 이러한 검토를 통해 연령 규범으로부터의 탈각을 위해 일본 정부가 도입하고 있는 고연령자 고용 정책의 의의와 한계에 대해 고찰하였다본 연구는 JSPS KAKENHI JP19K13480의 조성을 받아 수행한 연구임

Comparative expression of matrix-associated genes and inflammatory cytokines-associated genes according to disc degeneration: analysis of living human nucleus pulposus

STUDY DESIGN: The 2 groups of living human nucleus pulposus were prospectively compared according to disc degeneration. OBJECTIVES: This study was conducted to investigate the expressions of various genes associated with matrix synthesis and expressions of inflammatory cytokines-associated genes according to degrees of disc degeneration in human discs. SUMMARY OF BACKGROUND DATA: Degenerated discs were obtained from 18 patients who underwent discectomy for lumbar disc herniation. Disc degeneration was graded by T2-weighted magnetic resonance imaging using Pfirrmann's grading system. Discs were allocated to 2 groups: group I (9 patients)-mildly degenerated discs (grades II and III) and group II (9 patients)-severely degenerated discs (grades IV and V). METHODS: Cells from the nucleus pulposus were isolated and then cultured as monolayers. The mRNA expressions of aggrecan, type II collagen, Sox9, type I collagen, alkaline phosphatase, osteocalcin, tumor necrosis factor-α, and interleukin-1β in the 2 groups were compared by real-time polymerase chain reaction, and production of matrix-associate proteins (aggrecan, type II collagen, Sox9, type I collagen, alkaline phosphatase, and osteocalcin) were compared by Western blotting. RESULTS: mRNA expressions in group I were upregulated versus group II to the following extents: 1.83 times for aggrecan, 1.82 times for type II collagen, 1.80 times for Sox9, 1.41 times for type I collagen, 1.38 times for alkaline phosphatase, and 1.80 times for osteocalcin. Furthermore, Western blotting showed that aggrecan, type II collagen, Sox9, type I collagen, alkaline phosphatase, and osteocalcin were higher in group I. However, the mRNA levels of tumor necrosis factor-α and interleukin-1β were 1.26 and 1.11-fold, respectively, upregulated in group II. CONCLUSIONS: Mildly degenerated discs showed greater matrix, chondrogenic, and osteoblastic gene expressions than severely degenerated discs, indicating that the ability to produce matrix-associated proteins is greater for cells in mildly degenerated than in severely degenerated discs. However, inflammatory cytokine genes associated with disc degeneration were expressed at higher levels in the severely degenerated group. This study shows that a reduction in matrix synthesis and an increase of inflammatory cytokine levels occurs during disc degeneration at the same timeope

Histopathological Studies on Endometrial Hyperplasia and Carcinoma

A histopathological study was made on 65 cases of endometrial hyperplasia and endometrial carcinoma, that were obtained from the files of Departments of Pathology, College of Medicine, Seoul National University, and National Medical Center, during a 5-year period from 1975 to 1979_ Total 45 cases of endometrial hyperplasia were reclassified into endometrial hyperplasia without atypia (36 cases) , endometrial hyperplasia with minimal atypia (5 cases) , and endometrial hyperplasia with moderate atypia (3 cases) ,and endometrial hyperplasia with severe atypia(1 cases). Their ages ranged from 28 to 62, with a median of 46 years. Twenty cases of endometrial adenocarcinoma were reclassified into grade I (14 cases) ,grade II (3 cases) , and grade III (3 cases). The ages ranged from 28 to 71, with a median of 53.6 years

Borderline Lesion(Atypical Epithelial Lesion) of the Stomach

atypical epithelial lesion of the stomach were examined with authors' modified histo-topographic reconst· ruction method, and their pathologic characteristics were discussed along with its histogenesis. Five out of 7 cases were operated for either early or advanced carcinoma of the stomach, and only two were preoperatively diagnosed by endoscopic biopsy and radiological studies. Average age of the cases was 50.6 years, and majority were found in lower portion of stomach and mostly single in number. The lesions were all under 2.0 em in diameter, ranging from O. 5 em to 2. Oem. Macroscopically, they were sessile, poly paid, global and often mulberry-shaped, but 3 were made of flat and/or slightly depressed lesions. Cellular atypism of grade 1 comprised the bulk, with more prominent nuclear stratification and mitotic figures in the basal portion where they accompanied verrucous gastritis. cystic dilatation or intestinal metaplastic changes of proper glands as their underlying disorders below the lesion. And adjacent gastric mucosa represented a diffuse form of severe metaplastic gastritis. Those figures support that the borderline lesion may develop as a sequential step of dedifferentiated metaplastic gastritis during a reparative process of repeated mucosal erosions of the chronic gastritis

Which iodinated contrast media is the least cytotoxic to human disc cells?

BACKGROUND: Iodinated contrast media (CM) is commonly used for various intradiscal injections such as in discography and endoscopic spinal surgery. However, CM has been shown to be toxic to renal tissue due to its ionic strength and osmolarity and as a result of iodine-induced cytotoxicity, which has raised concern over whether there are similar negative effects on disc cells. PURPOSE: This in vitro study was designed to identify the least cytotoxic iodinated CM to the human disc cell among four different physiochemical iodinated contrast dyes. STUDY DESIGN: In vitro laboratory study. METHODS: Intervertebral disc tissue was obtained by discectomy from a total of 10 lumbar disc patients undergoing surgery and disc cells were isolated. The human disc cells were grown in 3D alginate bead culture with 0, 0.1, 10, and 100 mg/mL CM solutions (ionic dimer, ionic monomer, non-ionic dimer, and non-ionic monomer) and mannitol as a control for 2 days. The living cells were analyzed with trypan blue staining. Fluorescence-activated cell sorting analysis was performed using Annexin V and propidium iodide (PI) and 3D alginate bead immunostaining to identify live, apoptotic, and necrotic cells. RESULTS: Human disc cell death was time- and dose-dependent in response to CM and more necrosis was observed than apoptosis. In addition, non-ionic dimeric CM (iodixanol) showed the least toxic effect on human disc cells, followed by non-ionic monomeric (iopromide), ionic dimeric (ioxaglate), and ionic monomeric CM (ioxithalamate). CONCLUSIONS: Contrast media is cytotoxic to human disc cells in a dose- and time-dependent manner. This in vitro study revealed that, among four different CM preparations, non-ionic dimeric CM is the least detrimental to human disc cell viability. Careful attention should be paid to the type of CM chosen for discography and endoscopic spinal surgery. It is also necessary to investigate the detrimental effects of CM on disc cells and disc degeneration in further in vivo studies.ope

Time- and dose-dependent cytotoxicities of ioxitalamate and indigocarmine in human nucleus pulposus cells

BACKGROUND CONTEXT: Ioxitalamate (Telebrix 300) is an ionic iodinated contrast medium commonly used for discography or percutaneous endoscopic lumbar discectomy (PELD), though it has side effects such as anaphylactic shock and renal toxicity. Indigocarmine is an organic compound dye with a distinctive blue color that is commonly used during PELD to stain the acidic, degenerated nucleus pulposus (NP). Although ioxitalamate and indigocarmine are widely used in spinal surgery, there have been no reports on their effects on NP cells. We studied the toxicities of both ioxitalamate and indigocarmine to NP cells. PURPOSE: To determine the toxicities of both ioxitalamate and indigocarmine to NP cells in vitro. STUDY DESIGN: In vitro, controlled study of the toxicities of both ioxitalamate and indigocarmine to human NP cells. METHODS: Nucleus pulposus cells were obtained via discectomy from lumbar disc patients and isolated. Nucleus pulposus cells were cultured in three-dimensional (3D) alginate beads with 0.001, 0.1, 10, and 100 mg/mL ioxitalamate, 0.00001, 0.001, 0.1, and 10 mg/mL indigocarmine, or a mixture of both for 1, 2, or 3 days. The living cells were analyzed with trypan blue staining. Fluorescence Activated Cell Sorting analysis using Annexin V and propidium iodide and 3D alginate bead immunostaining was performed to identify live, apoptotic, and necrotic cells. RESULTS: Ioxitalamate, indigocarmine, and their combination induced statistically significant NP cell injury that was both time- and dose dependent (p<.05). Also, at the same concentration, ioxitalamate was more cytotoxic than was indigocarmine or the combination (p<.05). All three treatments also showed dose-dependent cytotoxicity according to flow cytometry and immunostaining. CONCLUSIONS: Ioxitalamate and indigocarmine are toxic to human NP cells in vitro in a time- and dose-dependent manner. We assume that ioxitalamate and indigocarmine may have similar effects in patients undergoing discography and PELD. Thus, we suggest that ioxitalamate and indigocarmine should be used carefully at low concentrations.ope