Mechanism of Honghua Oral Liquid in Alleviating Neuropathic Pain / 中国实验方剂学杂志

ObjectiveTo investigate the pharmacodynamic characteristics and explore the molecular mechanism of Honghua oral liquid （HOL） in relieving neuropathic pain （NP）. MethodHealthy male SD rats were randomly assigned into sham group， model group， low-， medium-， high-dose （0.5， 1.0， 2.0 mL·kg-1·d-1， respectively） HOL groups， and a positive drug （pregabalin， 25 mg·kg-1·d-1） group， with 6 rats in each group. Spinal nerve ligation （SNL） of L5 was conducted in other groups except the sham group. Drug administration was performed 3 days after the SNL surgery for 2 consecutive weeks， and samples were collected after the end of the administration. During the treatment period， the mechanical pain threshold and cold pain threshold were determined to measure the pain-relieving effect of HOL. Transcriptome sequencing was performed on hippocampal tissue samples from the sham， model， and high-dose HOL groups， and differentially expressed genes between the sham group and the model group as well as the model group and HOL high-dose group were obtained. After pathway enrichment analysis， we selected the targets which were closely related to neuroinflammation for validation， and predicted the specific binding sites of the major active components in HOL with the targets through molecular docking. In addition， the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） were determined by enzyme-linked immunosorbent assay （ELISA） to evaluate the effect of HOL on neuroinflammation in NP rats. ResultCompared with the sham group， SNL decreased the mechanical pain threshold and cold pain threshold （P<0.05）. Compared with the model group， HOL recovered the mechanical pain threshold and cold pain threshold （P<0.05）. The transcriptome data showed that 376 differentially expressed genes （DEGs） were identified between the model group and the sham group， including 124 upregulated genes and 252 downregulated genes， and 194 DEGs between the model group and the high-dose HOL group， including 33 upregulated genes and 161 downregulated genes. Among them， insulin-like growth factor 1（IGF1）， matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-14 （MMP-14）， erb-B2 receptor tyrosine kinase 2 （ERBB2）， and integrin subunit alpha 5 （ITGA5） associated with NP were selected for further validation. The Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） results showed that compared with the sham group， the modeling up-gurelated the mRNA levels of the above five molecules in the hippocampus （P<0.01）. Compared with model group， HOL down-regulated the mRNA levels of these molecules （P<0.01）. The molecular docking results showed that the main active components of safflower， hydroxysafflor yellow A， kaempferol， and quercetin， formed stable hydrogen bonds with the amino acid residues of IGF1， MMP-2， MMP-14， ERBB2， and ITGA5. The enzyme-linked immunosorbent assay（ELISA） results showed that compared with those in the sham group， the serum levels of TNF-α and IL-10 were out of balance in the model rats （P<0.01）. Compared with the model group， HOL lowered the level of the pro-inflammatory cytokine TNF-α （P<0.01） and elevated that of the anti-inflammatory cytokine IL-10 （P<0.05）. ConclusionHOL exerts analgesic effect on SNL rats by inhibiting neuroinflammation.

Microarray analysis of the relationship between PTEN and prognostic factors in primary breast cancer tissues / 肿瘤

Objective: To construct tissue microarray of breast carcinoma and to investigate the association of hepatocyte-specific phosphatase and tensin homolog (PTEN) protein expression with prognostic factors such as estrogen receptor (ER), progesterone receptor (PR), c-erb B-2, and Ki-67 in primary breast carcinoma. Methods: Tissue microarray of breast carcinoma was constructed. Immunohistochemical staining was used to detect the expression of PTEN, ER, PR, c-erb B-2, and Ki-67 inbreast carcinoma tissues. Results: PTEN was highly expressed in 66 out of 163 breast carcinoma tissues (40.5%). The positive rate of ER, PR, c-erb B-2, and Ki-67 was 51.5%, 35.6%, 34.4%, and 44.8%, respectively. The positive expression level of MN decreased with increase in auxiliary lymph nodes metastasis, histological grading, and TNM staging. They had negative correlations (P < 0.05). The positive expression level of PTEN had no evident correlation with tumor size and onset age. PTEN had positive correlation with the expressions of ER and PR protein (P < 0.01), and had negative correlation with the expressions of c-erb B -2 and Ki-67. Conclusion: There exists some relationships between low or loss of PTEN expression and the imbalanced expression of ER and PR and the growth and prognosis of breast carcinoma. PTEN may be considered as a potential indicator to judge the malignant degree of breast carcinoma. Combined detection of PTEN, ER, PR, c-erb B-2 and Ki-67 have significant importance and practical value for adjuvant therapy of breast carcinoma.

Análisis inmunohistoquímico de P53, CERB-B2, EGFR y PCNA en cáncer gástrico

Diversas anormalidades genéticas han sido reportadas en el desarrollo del carcinoma gástrico. Este trabajo estudia la inmuno expresión de marcadores inmunohistoquímicos: p 53, C-erb-B2, PCNA y EGFR, en 65 casos (37 hombres y 28 mujeres) con edad promedio de 54.5 años con cáncer gástrico precoz y avanzado. Histológicamente de acuerdo a la clasificación Japonesa, 8 casos (12,3%), se reportaron como adenocarcinomas moderadamente diferenciados, 7 (10.8%) como adenocarcinomas poco diferenciados con patrón sólido, 8 (12,3%) adenocarcinomas pobremente diferenciados patrón no sólido, 1 (1,5%) adenocarcinoma papilar, 12 (18,5%) carcinomas con células en anillo de sello y 12 (18,5%) carcinomas mucinosos. Tres lesiones (4,6%) fueron clasificadas como otro tipo de tumores, específicamente linfoepiteliomas. Según la clasificación de Lauren 23 lesiones (35.4%) se reportaron como carcinomas de tipo intestinal, 27 (41,5%) como carcinomas difusos, 1 (1.5%) como carcinoma mixto y en 14 casos (21.5%) los tumores se consideraron como no clasificables. Observándose: 37 casos positivos para p53, 24 con expresión de EGFR, 33 con expresión c-erbB2, y 55 que expresaron PCNA. La expresión de genes supresores, de factores de crecimiento, así como la expresión de marcadores de proliferación celular en el cáncer gástrico, son frecuentes en tumores gástricos avanzados y precoces En este estudio no se encontró correlación entre la expresión de los marcadores inmunohistoquímicos y los estadios clínico-patológicos.

Many genetic abnormalities have been reported in gastric cancer. We study the immunoexpression of p 53, Cerb B2, PCNA and EGFR in different types of gastric cancers. 65 cases (37 men and 25 women) with a rate of 54.5 years old with early and advanced gastric cancer from the records of our institute, 89.2% were advanced cases, 37cases positive to p53, 24 cases positive to EGFR and 55 cases positive to PCNA. Histological diagnosis was made according to the Japanese classification, 8 cases (12%) were moderately differentiated adenocarcinomas, 7(10.8%) poorly differentiated adenocarcinomas with a solid pattern type, 8(12.3%), poorly differentiated adenocarcinoma with a non solid pattern type 1(1.5%), papillary adenocarcinomas 12 (18.5%), signet-ring cell carcinoma and mucinous carcinoma 12 (18.5%). We also found 3(4.6%) lymphoepithelioma. According to Lauren's classification 23(35.4%) cases were intestinal type, 22(41.5%) cases diffuse type, 14(21.5%) not otherwise specified and 1 case mixed (intestinal and diffuse type). The expression of tumor Suppressors Genes, Growth Factors Genes and Proliferative Genes are frequently expressed in gastric cancer, but there are no correlations with clinical and pathological stages in this study.

The Relationship between Abnormal Screening Bone Scintigraphy and Bone Metastasis in Breast Cancer Patients / 한국유방암학회지

PURPOSE: In this study we evaluated the significance of false positive screening bone scintigraphy (BS) in primary invasive breast cancer patients. Lymphatic vessel invasion (LVI), estrogen receptor (ER), progesterone receptor (PR), nuclear grade, histology grade, epidermal growth factor receptor (EGFR) and C-erb-B2 values were examined in terms of their abilities to predict the accuracy of abnormal BS. We also examined the incidence of bone metastasis in primary invasive breast cancer patients according to the 1988 and 2003 AJCC classifications. METHODS: A retrospective review was performed on 2,044 primary invasive breast cancer patients that had received BS screening, and who were treated by mastectomy or breast conserving surgery at the Seoul National University Hospital between Jan 1995 and Jul 2003. Abnormal screening BS results were divided into "less suspicious" and "highly suspicious" groups. Patient's stages according to the 1988 AJCC classification were reclassified according to the 2003 AJCC classification. Bone metastasis was confirmed by further radiological examination or follow-up BS. All statistical analyses were two-tailed. RESULTS: The incidences of bone metastasis and an abnormal screening BS result were 1.7% (35/2,044) and 13.8% (283/2,044), respectively. The false positive rate of screening BS was 87.6% (248/283). LVI was the only significant predictive factor of bone metastasis in 283 of the abnormal BS patients (p <.001). c-erb-B2 showed no significance to predict bone metastasis in the "less suspicious" group, but was Bone is the most common site of distant metastasis in invasive breast cancer at the time of primary diagnosis. The vertebrae are the most common sites of bone metastasis and the ribs, skull, sternum and proximal long bones are also frequently involved. Bone metastases affect 8% of patients marginally significant in the "highly suspicious" group (p = .046). ER, PR, nuclear grade, histology grade, and EGFR showed no significance in terms of predicting the accuracy of an abnormal BS result. The incidences of bone metastasis were 0.6, 1.3 and 7.6% in stages I, II and III, respectively, according to the 1988 AJCC classification, while these incidences were 0.6, 0.7 and 5.8% according to the 2003 AJCC classification. CONCLUSION: The use of screening bone scintigraphy as a routine screening test is hard to justify due to its high false positive rate. LVI may be a useful factor in that it predicts the accuracy of an abnormal BS result. The incidences of bone metastasis in stages II and III were lower for the 2003 AJCC staging system.

Clinical Value of Cyclooxygenase-2 Expression in Human Breast Carcinoma / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: To determine whether COX-2 expression is associated with clinicopathological parameters, including c-erb-B2 overexpression and angiogenesis, and the disease- free survival of patients with operable breast cancer. MATERIALS AND METHODS: Paraffin-embedded tissue samples were selected from 205 patients surgically resected for breast cancer, between 1991 and 1997, and followed- up for at least 4 years. Samples were immunohistochemically stained with antibodies to COX-2, c-erb-B2 and CD34. RESULTS: COX-2 and c-erb-B2 expressions were detected in 118/205 (57.6%) and 58/205 (28.3%) patients, respectively. COX-2 expression was significantly higher in c-erb-B2 positive than c-erb-B2 negative tumors (75.9% vs. 49.7%, p-value 0.001). COX-2 expression was positively correlated with microvessel count (13.3+/-8.0 vs. 6.6+/-7.0, p-value 0.050), but not with other clinicopathological characteristics, including tumor size, involved axil lary lymph nodes and estrogen or progesterone receptor status. Although COX-2 expression itself was not a prognostic marker, breast cancer patients with tumors that co-expressed both COX-2 and c-erb-B2 had a poorer 5-year disease-free survival rate than those that did not (60.2% vs. 78.3%, p-value 0.0527). CONCLUSION: Our data suggest that COX-2 expression occurs frequently in c-erb-B2 positive breast cancer, and co-expression of COX-2 and c-erb-B2 may be a useful prognostic marker in patients with operable breast cancer.

Post-transcriptional control of c-erb B-2 overexpression in stomach cancer cells

The growth factor receptor oncogene, c-erb B-2, is frequently overexpressed in the adenocarcinomas of breast, ovary, lung and stomach. Although the mechanism of erb B-2 overexpression is thought as the result of transcriptional upregulation in many primary human carcinomas, expression rate of c-erb B-2 at mRNA level is usually lower than the level of translated protein. We also found that the expression of erb B-2 in SNU-1 stomach cancer cells was greater at post-transcription level (Bae et al., 1993). To explore the underlying mechanism of erb B-2 protein overexpression, we have chosen two cells lines, SNU-1 and SNU-16 where transcription rate of erb B-2 was closely resemble to each other while expressed protein levels were quite different. The synthesis rate of erb B-2 protein in SNU-1 cells was faster than SNU-16 cells while levels of erb B-2 mRNA were found to be similar in both cell lines. The half-life of the expressed erb B-2 protein was not significantly different in both cell lines. Analysis of the 5' untranslated region (UTR) of erb B-2 mRNA (-1approximately-323) showed no sequence abnormality in both cell lines. However, ribonuclease protection assay using cloned 5 UTR sequence revealed that the size of 5' UTR of erb B-2 mRNA which associate with transcription initiation site(s) in SNU-1 cells was longer than that in SNU-16. These results suggest that the increased erb B-2 protein synthesis rate possibly due to the redundant selection of transcription initiation might be a mechanism of erb B-2 overexpression in SNU-1 cells.

c-erb B2 and p53 Overexpression in Presence of Human Papilloma Virus Type 16/18 Infection in Carcinomas of the Uterine Cervix: Genesis and Clinicopathologic Analysis / 대한산부인과학회잡지

The 43 cases of the primary uterine cervical carcinoma were analyzed for HPV type 16/18 infection and also analyzed for overexpression of p53 and c-erb B2 oncoprotein to evaluate theirs oncogenic and clinicopathologic relationships. HPV type 16/18 infection was examined by polymerase chain reaction(PCR) and the overexpression of p53 and c-erb B2 protein by using immunohistochemical method. The results were as follow: 1. The HPV infection rate in primary uterine cervical carcinomas was 83.7% respectively.The standard clinicopathologic characteristics(age, histologic type, koilocytosis, mitosis, clinical stage, tumor size, cervical invasion depth, lymph node metastasis, parametrial invasion) were nat significantly correlated with HPV type 16/18 infectivity. 2. The overexpression rate of p53 protein was 72.1% and there was no Significant correlation between expression of p53 protein and the Clinicopathologic characteristics. 3. The overexpression of c-erb B2 oncoprotein was 44.2% and there was no significant correlation between the overexpression of c-erb B2 oncoprotein and the clinicopathologic characteristics. 4. There was no significant correlation between HPV type 16/18 infection and overexpression of p53 and c-erb B2 oncoprotein.

Expression of H-ras, erb B2, and p53 Proteins in Gastric Intestinal Metaplasia Associated with Cellular Atypism

Intestinal metaplasia (IM) have long been thought to play a role in the pathogenesis of gastric intestinal adenocarcinoma, but not in that of diffuse cancer. We studied 20 normal gastric mucosa, 90 IM, 39 atypia (dysplasia or adenoma), and 51 adenocarcinoma to evaluate the expression of p53, erb B2, and H-ras p21 proteins and to assess the correlation with IM (esp. type III IM, revealing positive HID-AB/PAS for sulfomucin). Positive rate of HID-AB staining revealed an increased trend in comparison between IM, atypia and adenocarcinoma. It was the highest in mucinous carcinoma, but it was not correlated with positive oncoprotein expressions. Positive rates of oncoproteins revealed increased trends in comparison between IM, dysplasia or adenoma and adenocarcinoma in c-erb B2 and p53 (P<0.01). The positive rates were highest in intestinal adenocarcinoma (50.0% and 54.2%, respectively). Rates were lowest in biopsy tissue of IM (4.4% and 8.7%, respectively). The expression of H-ras p21 was not significant in gastric carcinogenesis. There was no significant correlation between oncoproteins and other clinical parameters, such as depth of invasion, differentiation, size and nodal metastasis of the tumors. Therefore, we suggest that p53 and erb B2 may play a role in the carcinogenesis of gastric intestinal adenocarcinoma.

Cyto - molecular Biologic Characterization of c - myc , erb B and EGF - Receptor in Squamous Cell Carcinoma / 대한피부과학회지

BACKGROUND: Oncogenes and EGF-Receptor(EGFR) may be involved n different stages of the multistep carcinogenesis process. A specific pattern of karyotypic abnormalities in solid tumors can be detected by cytogenetic methods. OBJECTIVE: This study is intnded to observe the cytomolecular kiologic chracterization of c-myc, erb B and EGFR genes in squasnous cell carcinoma(SCC) of the skin and cervix. METHODS: We have eytogenet,ically examined the short-term culturs from SCC. The rearrangement, amplification or expressi.on of erb B, c-myc, and EGFR genes were studied by Southern blot, analysis of genomic DNA and by slot blot analysis of tota! RNA extracted from biopsies of normal skin and SCC tissues. EGFR expression was examined immunohistochemially using monoclonal antibodies and the localizat,ion of the c-myc oncogene mRNA by in situ hybridization. RESULTS: A remarkably structural aberration was del 6(q21-qter) counted 20 metaphases among 28 metaphases ana1yzed. In nunierical aberration, all chromosomes were lost or gained randomly. Amenploid including triploid and tetraploid were observed in 8 metaphases, 6 tumor cells contained marker chromosome. In Southern blot analysis, rearrangement and amglificaton of EGFR in primary squamous cell carcinoma of cervix uteri and skin respectively. In slot blot analysis, the levels of c-myc, erb B and EGFR mRNA increaaed respectively 3.5, 2.5 and 2.8 times in SCC when compared to normal tissues. In immunoperoxidase stain, EGFR was present, in SCC where keratinocytes with strong cyto-plasmic staining but no membr, line labelling, where as in normal skin the were primarily present in t,he membrane and cytoplasm of basal cells. In situ hybridization with c-myc cDNAs allowed detection of grains representative of biotin labelled cDNA-mRNA hybrids in the frozden section of SCC tissues. CONCLUSION: These results suggest that specific patterns of karyotypir abnormalites, rearrangement, or amplification of EGFR gene, and overexpression of oncogenes and EGFR gene may be associated with the carcinogenesis of SCC.