• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.3903-3909
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• 2016

Purpose: To evaluate the expression of Her2/neu and Ki-67 in benign and malignant gallbladder lesions, and to establish correlations with clinico-pathologic parameters. Materials and Methods: A retrospective analysis was conducted on formalin fixed paraffin embedded (FFPE) benign (n=25) and malignant gallbladder (n=25) tissue samples. Hematoxylin and eosin stained slides of each case were reviewed for: type of malignancy (whether adenocarcinoma, squamous cell carcinoma, or any other type), grade (well, moderate, and poor), depth of invasion, pre-neoplastic changes in adjacent mucosal epithelium like metaplasia and dysplasia. Immunohistochemistry for Her 2 neu and Ki-67 was performed and data analysis was conducted using SPSS 17 software. Chi-square test was used to compare categorical/dichotomous variables. P value of ${\leq}0.05$ was considered significant. Results: The difference of Her 2 neu expression and Ki67 index between benign and malignant groups was found to be statistically significant. Her2/neu positivity did not have any significant correlation with various clinicopathological parameters other than liver involvement. 5 cases of gallbladder cancer showed both Her2/neu and Ki67 positivity. Ten cases were Ki67 positive but Her2/neu negative while one case was Her2/neu positive but Ki67 negative. Conclusions: The present study demonstrated overexpression of Her2/neu and Ki67 in gallbladder cancer. A trend of decreasing Her2/neu expression with increasing grade of tumor was observed. Furthermore, greater Ki67 positivity was found in cases with lymph node metastasis and distant metastasis. Future studies with a larger number of patients will be required to precisely define the correlation of Her2/neu expression and Ki67 positivity with clinicopathological parameters. The results however are encouraging and suggest evaluation of Her2/neu as a candidate for targeted therapy.

• The Journal of Korean Physical Therapy
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• 제23권2호
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• pp.45-52
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• 2011

Purpose: The purpose of this study was to investigate the effect of electroacupuncture on NeuN expression in ventral horn motor neurons of spinal cord, changes in pain thresholdchanges in motor function in rats with partially dissected sciatic nerves. Method: A total of 120 male Sprague-Dawley rats were randomly divided into two groups, a control group and a group administered electroacupuncture at ST36, LI11 and SP9 with 120 Hz and 0.5 mA. Animals were sacrificed on days 1, 3, 7, 14 and 28 after nerve injury (the sciatic nerve was partially dissected). The pain threshold was recorded by an Analgesia? meter and a BBB? score was calculated for motor function. After preparing lumbar spinal cord slide sections, they were immunostained with NeuN antisera (1:2,500). Results: The numbers of NeuN immunoreactive neuronsin the electroacupuncture group was increased compared to the control group. The numbers of NeuN immunoreactive neurons on days 14 and 28 day were different (p<0.05), as were the numbers on days 3 and 7 (p<0.01). The pain threshold BBB score for the electroacupuncture group was higher than for controls. Conclusion: The increase in pain threshold, BBB-score and number of NeuN immunoreactive neurons inventral horn motor neurons of spinal cord in rats withnerve dissection showed that electroacupuncture can attenuate pain transduction and increase motor function. Also, NeuN was a good marker for identifying the degree of nerve cell loss after nervous system injury.

• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.3733-3736
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• 2016

HER2/neu is a well-established prognostic and predictive factor for invasive breast cancer. However, the role of HER2/neu in ductal breast carcinoma in situ (DCIS) is debated and recent data have suggested that it is mainly linked to in situ local recurrence. Although molecular data suggest that atypical ductal hyperplasia (ADH) and duct carcinoma in situ (DCIS) are related lesions, albeit with vastly different clinical implications, the role of HER2/neu expression in atypical ductal hyperplasia is not well defined either. The aim of this study was to evaluate over expression of HER2/neu in DCIS and cases of ADH in comparison with invasive breast carcinoma. Archival primary breast carcinoma paraffin blocks (n=15), DCIS only (n=10) and ductal epithelial hyperplasia and other breast benign lesions (n=25) were analyzed for HER2/neu immunoexpression. Follow up was available for 40% of the patients. HER2/neu was positive in 80%of both DCIS and invasive carcinoma, and 67% of atypical ductal hyperplasia (ADH) cases. Thus at least a subset of patients with preinvasive breast lesions were positive, which strongly suggests a role for Her2/neu in identifying high-risk patients for malignant transformation. Although these are preliminary data, which need further studies of gene amplification within these patients as well as a larger patient cohort with longer periods of follow up, they support the implementation of routine Her2/neu testing in patients diagnosed as pure DCIS and in florid ADH.

• Clinical and Experimental Pediatrics
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• 제49권5호
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• pp.558-564
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• 2006

목 적 : 경뇌 전자기 자극법은 변조 자기장을 이용하여 뇌세포에 대한 직접적인 영향을 주지 않으면서 중추 신경계를 자극할 수 있는 비침습적인 방법이다. 이전의 연구들은 대부분 생체 동물을 대상으로 수행되어져 왔으며 배양 조직에서의 연구는 별로 이루어진 바 없다. 이에 본 연구에서는 배양된 해마 절편에서 다른 주파수의 전자기 자극이 신경원에 미치는 영향과 약물에 의한 세포 손상 후 전자기 자극의 세포보호효과 여부에 대해 알아보고자 하였다. 방 법 : 생후 8일된 실험쥐의 대뇌를 적출하여 dissection microscope 하에서 양쪽 해마 부위를 분리하고 tissue chopper를 이용하여 $450{\mu}M$ 두께로 절편을 만든 후 Stoppini가 고안한 방법대로 배양을 시행하였다. 각각 5개의 건강한 해마 절편이 포함된 inserts를 선택하고, 전자기 자극군에 대해 0.67 Hz와 50 Hz의 주파수로 각각 배양 5일부터 3일 간격으로 6차례 전자기 자극을 가하였다. 또한, 배양 제 14일에 inserts 2개에 $100{\mu}M$ NMDA에 노출시키고 3일 후부터 3일 간격으로 insert 1개에 전자기 자극을 3차례 시행하였고 다른 1개의 insert와 대조군에는 자극을 가하지 않았다. 결 과 : 전자기 자극 후 신경원의 활성도를 알아보기 위해 NeuN 단백 발현을 western blotting을 이용하여 측정한 후 ${\beta}$-actin 단백 발현과의 비를 얻어 각 군에서 비교 분석하였다. 대조군($1.01{\pm}0.27$)에 비해 전자기 자극군에서 NeuN의 발현이 증가되어 있었으며, 특히 저주파 자극군($1.12{\pm}0.14$)에서보다 고주파 자극군($1.27{\pm}0.17$)에서 현저하였고 고주파 자극군에서는 대조군에 비해 통계적으로 유의한 증가를 보였다(P<0.05). 또한, NMDA 노출 후 실험군(전자기 자극군 : $1.15{\pm}0.27$, 전자기 비자극군 : $0.92{\pm}0.09$)에서 대조군($1.26{\pm}0.04$)에 비해 NeuN 발현이 감소되었으나 전자기 비자극군에서 더 많이 감소한 것을 알 수 있었다. 결 론 : 배양된 해마조직의 신경세포에 대한 전자기 자극은 저주파 자극군에 비해 고주파 자극군에서 대조군보다 통계적으로 유의한 수준의 NeuN 발현의 증가를 관찰할 수 있었고, NMDA 노출 후 전자기 자극을 가한 군에서 대조군보다 NeuN 발현 감소가 관찰되기는 하였으나 고주파 자극군에서는 통계적으로 유의하지 않은 정도였던 것으로 보아 전자기 자극이 신경원 활성을 증가시켜 신경세포의 발생 및 증식을 유도하며 세포 손상에 대한 신경보호효과도 보인다는 것을 알 수 있었다. 따라서 전자기 자극은 여러 신경 질환에 있어서 치료적 역할을 할 수 있는 가능성이 있다고 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권2호
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• pp.691-695
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• 2016

Protein glycosylation is the most common posttranslational modification in mammalian cells. Aberrant protein glycosylation has been reported in various diseases, including cancer. We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-$MS^n$). Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by $NeuA_{c2}Gal_1GalNA_{c1}$. These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.722-727
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• 2005

Cytosolic sialidase (Neu2), a member of the sialidase family that is responsible for hydrolysis of sialic acid from the terminal position of sialoglycoconjugates, is poorly expressed in skeletal muscle and not detected in any other adult tissues. Thus, we isolated Neu2 cDNA using splicing by overlap extension (SOEing). In order to further characterize this enzyme, a His-tagged derivative was expressed in the bacterial expression system and purified by $Ni^{2+}$-affinity chromatography. A recombinant product of approximately 42 kDa had sialidase activity toward 4-methyl-umbelliferyl-$\alpha$-D-N-acetylneuraminic acid (4MU-NeuAc). The optimal pH and temperature of the recombinant Neu2 for 4MU-NeuAc was 6.0 and $37.5^{\circ}C$, respectively. The metal ions, such as $Cu^{2+}\;and\;Cd^{2+}$, showed strong inhibitory effect on the activity of the enzyme. The enzyme efficiently hydrolyzed the gangliosides GM3 and GD3 and had relatively low activities on ganglioside GD1a and GD1b, $\alpha$2-3 sialyllactose, and sialylated glycoproteins such as fetuin, transferrin, and orsomucoid, but had hardly any activities on $\alpha$2-6 sialyllactose and ganglioside GM1 and GM2. We concluded that the recombinant Neu2 has a sialidase activity toward glycoproteins as well as gangliosides.

• Archives of Plastic Surgery
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• 제34권1호
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• pp.1-7
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• 2007

Purpose: Human adipose tissue-derived stromal cells(hADSCs) can be expanded in vitro and induced to differentiate into multiple mesenchymal cell types. In this study we have examined various neuronal phenotypes and gene expression profiles of the hADSCs in the neuronal induction. Methods: The hADSCs were isolated from human adipose tissue and they were characterized by the flow cytometry analysis using CD13, CD29, CD34, CD45, CD49d, CD90, CD105 and HLA-DR cell surface markers. We differentiated the hADSCs into the neuronal lineage by using chemical induction medium and observed the cells with contrast microscopy. The immunocytochemistry and western blotting were performed using the NSE, NeuN, Trk-A, Vimentin, N-CAM, S-100 and ${\beta}$-Tubulin III antibodies. Results: The hADSCs were positive for CD13($90.3{\pm}4%$), CD29($98.9{\pm}0.7%$), CD49d($13.6{\pm}6%$), CD90 ($99.4{\pm}0.1%$), CD105($96%{\pm}2.8%$) but negative for CD34, CD45 and HLA-DR. The untreated cultures of hADSCs predominately consisted of spindle shaped cells and a few large, flat cells. Three hours after the addition of induction medium, the hADSCs had changed morphology and adopted neuronal-like phenotypes. The result of immunocytochemistry and western blotting showed that NSE, NeuN, Trk-A, Vimentin, N-CAM, S-100 and ${\beta}$-Tubulin III were expressed. However, NSE, NeuN, Vimentin were weakly expressed in the control. Conclusion: Theses results indicate that hADSCs have the capabillity of differentiating into neuronal lineage in a specialized culture medium. hADSCs may be useful in the treatment of a wide variety of neurological disorders.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6311-6318
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• 2012

Background: HER2/neu overexpression due to gene amplification is an important factor in breast cancer, modifying the sensitivity to anti-HER2 monoclonal antibody therapy. The clinical significance of HER2 expression in non small cell lung carcinoma (NSCLC) is currently under evaluation. The tumor suppressor gene PTEN negatively regulates the HER2/PI3K/Akt signalling pathway. The purpose of this study was to evaluate the role of simultaneous alteration in HER2 and PTEN protein expression in relation to biological behaviour of NSCLCs. Materials and Methods: Protein expression was determined by immunohistochemistry in sixty-one (n=61) NSCLC cases along with CISH for HER2 gene analysis and detection of chromosome 17 aneuploidy. Patients were followed-up for a period of 34 to 41 months after surgery. Results: HER2 overexpression (2+/3+score) was detected in 17 (27.9%) patients while loss of PTEN expression was observed in 24 (39.3%) cases, low expression in 29 (47.6%) and overexpression in 8 (13.1%). Simultaneous HER2 overexpression and PTEN low/loss of expression were correlated with metastasis (71.4% vs 36.2% p=0.03). Analysis in the subgroup of 22 patients of pTNM stage III with lymph node status N1 or N2 revealed that there was a relationship between the number of positive regional lymph node groups and simultaneous deregulation of the two genes (p=0.04). Multivariate analysis determined that HER2 overexpression was associated with an increasing risk of developing metastases (OR: 4.3; 95%CI: 1.2-15.9; p: 0.03) while PTEN overexpression was associated with lower risk (OR: 0.1; 95%CI: 0.1, 1.0; p: 0.05). Conclusions: Simultaneous HER2/PTEN deregulation is a significant genetic event that leads to a more aggressive phenotype of NSCLC.

• 대한의생명과학회지
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• 제21권2호
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• pp.92-102
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• 2015

This work aimed to study whether rice bran extract fermented with Lactobacillus plantarum (LW) promotes functional recovery and reduces cognitive impairment after ischemic brain injury. Ischemic brain injury was induced by middle cerebral artery occlusion (MCAO) in rats. Four groups were studied, namely the (1) sham, (2) vehicle, (3) donepezil, and (4) LW groups. Animals were injected with LW once a day for 7 days after middle cerebral artery occlusion. LW group showed significantly improved neurological function as compared to the vehicle group, as well as enhanced learning and memory in the Morris water maze. The LW group showed the greatest functional recovery. Moreover, the LW group showed an enhanced more survival cells anti-apoptotic effect in the cortex and neural cell densities in the hippocampal DG and CA1. In addition, this group showed enhanced expression of neurotrophic factors, antioxidant genes, and the acetylcholine receptor gene, as well as synaptophysin (SYP), Fox-3 (NeuN), doublecortin (DCX), and choline acetyltransferase (ChAT) proteins. Our findings indicate that LW treatment showed the largest effects in functional recovery and cognitive improvement after ischemic brain injury through stimulation of the acetylcholine receptor, antioxidant genes, neurotrophic factors, and expression of NeuN, SYP, DCX, and ChAT.

• 한국발생생물학회지:발생과생식
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• 제2권2호
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• pp.149-155
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• 1998

폴리시알린산 (polysialic acid)은 미생물로부터 인간에 이르기까지 다양한 세포의 세포 표면 복합당사슬에 공유 결합하는 당사슬의 일종이다. 최근 성게 난자의 젤리층과 성게 난자의 표면에서 폴리시알린산이 연결된 당단백질이 발견됨으로써 폴리시알린산의 생합성과 그 기능에 대한 의문이 제기되었다. 둥근성게의 난자와 배에서 추출한 효소액과 CMP-[$^{14}$ C]Neu5Ac를 기질로 사용하여 Neu5Ac를 endogenous acceptor에 전달하는 CMP-Neu5Ac:poly-$\alpha$2, 8 sialosyl sialyltransferase (polyST)를 발견하였다. polyST는 20mM MOPS (pH 7.0), 2$0^{\circ}C$에서 최적 활성을 나타냈다. 10 mM $Mg^2$$^{+}$를 효소 반응액에 첨가하면 polyST의 황성은 2.7배 증가하였다. poiyST는 또한 포유류의 gang1ioside인 GD3에 폴리시알린산을 합성할 수 있었다. 이와 같은 폴리시알린산이 존재하는 ganglioside가 현재까지 자연계에서 알려진 바가 없다는 사실은 하나의 polyST가 endogenous acceptor와 ganglioside에 폴리시알린산을 합성했다고 결론지을 수 있다. 과량의 GD3를 exogenous acceptor로 사용하여 둥근성게에 존재하는 polyST의 활성을 조사한 결과 mesenchyme 포배기 때부터 그 활성이 급격히 증가하고 낭배기 때에 최고에 도달했다. 성게의 배발생 때에 polyST의 활성이 조절된다는 것은 낭배기 때에 일어나는 세포나 조직 사이에 일어나는 변화와 초기 단계 spicule 형성에 어떤 중요한 기능을 담당하고 있을 가능성이 있다.