Chemical Composition of Artemisia argyi Extract (RW0117) and Protective Effects against Gastric Lesions in vivo

In this study, we investigated the chemical profile and effects of RW0117 (Artemisia argyi 65 .5 % ethanol extract) on gastric lesions in rats. We optimized and validated a method to obtain the chemical profile of RW0117. We then investigated the antioxidant and anti-inflammatory effects in vivo, and the protective effects on gastric lesions in vivo. The IC50 of 2,2-diphenyl-1-picrylhydrazyl free radical scavenging considering the antioxidant effects of RW0117 was 166.55 μg/mL, and the IC50 of nitric oxide scavenging considering the antiinflammatory effects was 41.16 μg/mL. Oral administration of RW0117 at lower concentrations (25, 50, 100 mg/ kg) had similar or greater effects than the daily intake conversion concentration (115mg/kg) of a health functional food (Avexol® ) in the acetic acid-induced ulcer and the ethanol-induced gastric injury rat models. In addition, oral administration of RW0117 increased the expression of prostaglandin E2 , which enhances the protective effect in the gastric mucosa in the ethanol-induced gastric injury rat model. These results suggest that RW0117 may have potential therapeutic uses in the protection of the gastric mucosa.

Chemical Composition of Artemisia argyi Extract (RW0117) and Protective Effects against Gastric Lesions in vivo

In this study, we investigated the chemical profile and effects of RW0117 (Artemisia argyi 65 .5 % ethanol extract) on gastric lesions in rats. We optimized and validated a method to obtain the chemical profile of RW0117. We then investigated the antioxidant and anti-inflammatory effects in vivo, and the protective effects on gastric lesions in vivo. The IC50 of 2,2-diphenyl-1-picrylhydrazyl free radical scavenging considering the antioxidant effects of RW0117 was 166.55 μg/mL, and the IC50 of nitric oxide scavenging considering the antiinflammatory effects was 41.16 μg/mL. Oral administration of RW0117 at lower concentrations (25, 50, 100 mg/ kg) had similar or greater effects than the daily intake conversion concentration (115mg/kg) of a health functional food (Avexol® ) in the acetic acid-induced ulcer and the ethanol-induced gastric injury rat models. In addition, oral administration of RW0117 increased the expression of prostaglandin E2 , which enhances the protective effect in the gastric mucosa in the ethanol-induced gastric injury rat model. These results suggest that RW0117 may have potential therapeutic uses in the protection of the gastric mucosa.

Histopathologic Analysis of BI-RADS Category 4a Breast Lesions Diagnosed by Ultrasonography / 한국유방암학회지

PURPOSE: We analyzed the histopathologic findings of the patients with ultrasongraphic Breast Imaging Reporting and Data System (BI-RADS) Category 4a breast lesions to determine which patient can be excluded from any invasive, diagnostic procedure in the future. METHODS: Of the 180 cases of BI-RADS Category 4a breast lesions that were diagnosed with ultrasonography during a 6 month-period, 132 cases were pathologically confirmed and these were analyzed retrospectively. Four benign cases that did not undergo any further procedure after fine needle biopsy and 6 malignant cases (4.5%) were excluded from this study. RESULTS: Of the 122 cases, 77 cases (63.1%) showed homogeneous benign finding, and 45 cases (36.9%) showed heterogeneous finding that was made up of two or more different pathologic lesions. Fibroadenoma (55.8%) was the most frequent pathologic finding in the cases with homogeneous finding, followed by fibrocystic change (14.3%), and fibrosis (7.8%). The cases with heterogeneous finding presented fibrocystic change (55.5%), microcalcification (48.8%), ductal hyperplasia (42.2%), and fibroadenoma (31.1%) in the order of frequency. CONCLUSION: Lesion with heterogeneous histopathologic nature was the most frequent finding defined as category 4a in breast ultrasonography, followed by fibrodenoma, fibrocystic change, microcalcification, and ductal hyperplasia. Refining more specific ultrasonographic findings of these lesions would guarantee that radiologists exclude more benign lesions from category 4a.

Expression of the Markers for the Mammary Stem Cells and the Cellular Lineages of the Mammary Gland on Ductal Carcinoma In Situ / 한국유방암학회지

PURPOSE: Breast Cancer is an inter-tumoral and intra-tumoral heterogeneous disease. It remains unclear whether this heterogeneity results from different target cells or from different subsets of genetic abnormalities, otherwise from both. We postulated that in addition to genetic cloning, a variety of cells that exist during the defined developmental stages of the human mammary gland could give rise to the heterogeneity of breast cancer. To verify this postulation, we have analyzed pure ductal carcinoma in situ (DCIS) for the expression of the biomarkers that represent the mammary stem cell, the early progenitor cells, and the glandular and myoepithelial cells of the mammary gland. METHODS: We investigated the relationship between the immnuohistochemical expression of the mammary development-associated biomarkers {cytokeratin-18 (CK18), cytokeratin-6 (CK6), alpha-smooth muscle actin (SMA), Wnt-1, Notch 3} and some other factors {the menopausal status, the estrogen receptor (ER) status, the progesterone receptor (PR) status, c-erbB-2, and the number of tumor foci} in 26 cases of DCIS. RESULTS: All 26 cases included in this study showed the positive expressions of CK18 and SMA. The expression of all the markers was not correlated with the menopausal status. The positive expression of CK6 had a statistically significant relationship with a negative estrogen receptor (p=0.014), positive c-erbB-2 (p=0.048), high nuclear grade (p=0.001), and single focus of DCIS (p=0.017). The expression of Wnt-1 and Notch 3 did not have significant correlation with any factors. However, the positive expression of Wnt-1 showed a tendency of a negative ER (p=0.061) and the positive expression of Notch 3 also showed a tendency of a negative ER (p=0.086) and a high nuclear grade (p=0.086). CONCLUSION: The CK6 positive tumor is thought to originate from the more primitive cells compared to the CK6 negative tumor. Unifocality of the CK positive tumor might result from the arrest of differentiation of the original cell after disease affection. DCISs could be categorized into the CK6 positive and negative groups.

Detection of micrometastasis in fixed paraffin-embedded Sentinel Lymph Nodes of Breast cancer using RT-PCR / 한국유방암학회지

PURPOSE: Sentinel lymph node (SLN) biopsy is considered a highly accurate and very economic method of assessing the axillary nodal status in breast cancer patients. Recently immunohistochemical (IHC) staining and reverse transcriptase polymerase chain reaction (RT-PCR) are commonly used to evaluate micrometastasis in the sentinel lymph node. However, most of the RT-PCR studies have been performed using fresh tissue. This study was conducted to assess micrometastasis in clinically node-negative breast cancer by using RT-PCR technique on the paraffin embedded sentinel lymph nodes. METHODS: Sixty patients who undergone SLN biopsy followed by axillary lymph node dissection due to breast carcinoma were evaluated from February 2000 to January 2001 at the Breast Cancer Center, Department of Surgery, Yongdong Severance Hospital. Serial sections were made from all sentinel lymph nodes for the H&E staining and for the IHC staining with monoclonal anti-cytokeratin antibody. RNA was extracted from the paraffin embedded sentinel lymph nodes and RT-PCR was performed using cytokeratin 19 mRNA, MUC-1 mRNA, and MAGE-A3 mRNA. RESULTS: In 32 out of 60 cases, beta-actin mRNA was detected after RT-PCR, and the 28 cases which had no product after RT-PCR for beta-actin were excluded from this study. Twenty five cases showed as being metastasis positive and 7 cases showed as being metastasis negative by serial section (SS) H&E staining. Three out of 25 negative cases tested for by SS H&E staining were found to be positive by IHC. Ten, six and, eight cases out of the 25 negative cases tested for by SS H&E were found to be positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Among the 22 cases that were found to be negative by both SS H&E staining and IHC staining, 9, 4, and 6 cases were converted to positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Using the combination of two or three markers for performing RT-PCR was more sensitive than any single marker to detect micrometastasis (p < 0.05). CONCLUSION: Even though we failed to extract RNA in 46% of the paraffin embedded tissues, it may be possible to detect micrometastasis by using RT-PCR with the paraffin embedded tissue. RT-PCR is far more sensitive than IHC for detecting microme tastasis, and when we combine multiple markers, the detection rate is higher than for any one marker.

Detection of micrometastasis in fixed paraffin-embedded Sentinel Lymph Nodes of Breast cancer using RT-PCR / 한국유방암학회지

PURPOSE: Sentinel lymph node (SLN) biopsy is considered a highly accurate and very economic method of assessing the axillary nodal status in breast cancer patients. Recently immunohistochemical (IHC) staining and reverse transcriptase polymerase chain reaction (RT-PCR) are commonly used to evaluate micrometastasis in the sentinel lymph node. However, most of the RT-PCR studies have been performed using fresh tissue. This study was conducted to assess micrometastasis in clinically node-negative breast cancer by using RT-PCR technique on the paraffin embedded sentinel lymph nodes. METHODS: Sixty patients who undergone SLN biopsy followed by axillary lymph node dissection due to breast carcinoma were evaluated from February 2000 to January 2001 at the Breast Cancer Center, Department of Surgery, Yongdong Severance Hospital. Serial sections were made from all sentinel lymph nodes for the H&E staining and for the IHC staining with monoclonal anti-cytokeratin antibody. RNA was extracted from the paraffin embedded sentinel lymph nodes and RT-PCR was performed using cytokeratin 19 mRNA, MUC-1 mRNA, and MAGE-A3 mRNA. RESULTS: In 32 out of 60 cases, beta-actin mRNA was detected after RT-PCR, and the 28 cases which had no product after RT-PCR for beta-actin were excluded from this study. Twenty five cases showed as being metastasis positive and 7 cases showed as being metastasis negative by serial section (SS) H&E staining. Three out of 25 negative cases tested for by SS H&E staining were found to be positive by IHC. Ten, six and, eight cases out of the 25 negative cases tested for by SS H&E were found to be positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Among the 22 cases that were found to be negative by both SS H&E staining and IHC staining, 9, 4, and 6 cases were converted to positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Using the combination of two or three markers for performing RT-PCR was more sensitive than any single marker to detect micrometastasis (p < 0.05). CONCLUSION: Even though we failed to extract RNA in 46% of the paraffin embedded tissues, it may be possible to detect micrometastasis by using RT-PCR with the paraffin embedded tissue. RT-PCR is far more sensitive than IHC for detecting microme tastasis, and when we combine multiple markers, the detection rate is higher than for any one marker.

Predicting the Status of the Nonsentinel Lymph Nodes in early Breast Cancer Patients with Positive Sentinel Lymph Nodes / 한국유방암학회지

PURPOSE: The need for completion axillary lymph node dissection, even in early breast cancer patients with a positive sentinel lymph node, has been questioned. The purpose of this study was to determine the factors that predict the presence of metastasis in non-sentinel lymph nodes (NSLNs) when the sentinel lymph node (SLN) was positive. METHODS: Between December 1998 and June 2004, the records of 104 early breast cancer patients with a positive SLN and who underwent completion axillary lymph node dissection were reviewed. The clinicopathological features in SLN-positive patients were evaluated as possible predictors of metastatic NSLN. RESULTS: Forty four (42.3%) of the 104 patients with positive SLN had metastatic NSLNs. In a univariate analysis, unicentric multifocality (P=0.016), lymphovascular invasion (P=0.006) and SLN metastasis larger than 2 mm (P= 0.003) were associated with positive NSLN findings. The number of SLNs removed was significantly associated as a negative predictor (P=0.043). A multivariate analysis revealed that SLN metastasis >2 mm (P=0.021) and lymphovascular invasion (P=0.040) were independent predictors of metastatic NSLN. CONCLUSION: The likelihood of metastatic NSLNs correlates with the size of the largest SLN metastasis and the presence of lymphovascular invasion of the primary tumor. Even though in early breast cancer with positive SLNs, incorporating these factors may help determining which patients would benefit from additional axillary lymph node dissection.

Intraoperative Examination of Sentinel Lymph Nodes Using Rapid Cytokeratin Immunohistochemical Staining in Breast Cancer Patients / 한국유방암학회지

PURPOSE: Sentinel lymph node (SLN) biopsy has become a new standard procedure in the treatment of patients with early breast cancer. Furthermore, many institutions have begun offering the sentinel lymph node biopsy without simultaneous axillary dissection as a possible standard procedure when the SLN was free from tumors. For appropriate intraoperative decision making on the presence of cancer cells in axillary lymph nodes, a fast and accurate method to assess the SLN is required. The authors performed a prospective investigation of the relative merits of rapid cytokeratin immunohistochemical (IHC) staining of the SLN removed during the operations of breast cancer patients. METHODS: Between December 2002 and August 2003, 38 patients with T1and T2 breast cancer were enrolled after undergoing successful sentinel lymph node biopsy. A total of 60 sentinel lymph nodes (mean number, 1.58) were biopsied and first examined by hematoxylin-eosin (H&E) staining. All the tumor free sentinel lymph nodes by H&E stained section were immunostained for cytokeratin using a rapid immunohistochemical assay (Cytokeratin (PAN), 1:50, Newcastle, UK) during the operation. Finally, the sentinel lymph nodes were submitted for paraffin embedding and serial section after surgery. Both H&E stained and cytokeratin immunostained sections were also performed. RESULTS: This technique has a turnaround time of less than 20 minutes during the operation. Rapid IHC staining revealed 4 positive sentinel lymph nodes that were negative for metastasis by H&E staining. Among these false negative 4 cases, two cases had problems with the frozen section of H&E staining and the other 2 cases had problems due to micrometastasis. This study showed a sensitivity of 88.89%, a specificity of 100%, an accuracy of 98.33%, and a negative predictive value of 98.08%. The false-negative case (1 of 52), which was negative on H&E staining and rapid IHC staining during the operation, was disclosed as positive only after a serial permanent section examination with IHC stain. CONCLUSION: The introperative examination of sentinel lymph nodes is a highly accurate and effective way of predicting the axillary lymph node status of patients with breast cancers. This may be a promising technique in deciding whether to spare axillary lymph node dissection for the patient in the operating room.

Clinical Experience for Sentinel Lymphadenectomy Alone in Early Breast Cancer / 한국유방암학회지

PURPOSE: This study was performed to evaluate the efficacy and safety of sentinel lymph node biopsy alone (SLNB) without axillary lymph node dissection. METHODS: Between Jun. 1999 and Dec. 2002, we carried out SLNB in 85 consecutive patients with T1 breast cancer whose sentinel lymph node(s) were tumor-free on intra- operative frozen section. Even when sentinel lymph node(s) turned out to be positive by permanent pathology, additional axillary lymph node dissection was not performed. Patients underwent total mastectomy or partial mastectomy and received an appropriate adjuvant therapy according to the characteristics of the primary tumor. All patients who had breast conserving surgery received postoperative radiotherapy to the remaining breast, but not to the axilla. RESULTS: SLNB only took 14 minutes and yielded no postoperative complications. Among the 85 patients whose sentinel lymph nodes were tumor-free on frozen section, 11 patients were diagnosed as metastatic in the permanent pathology. One of them had a macro-metastasized (2.5 cm) sentinel lymph node, and the rest had micro-metastasized sentinel lymph nodes. For 23.1 months of mean follow-up period, all the patients including a patient who died of liver metastases at 17 months showed no evidence of axillary recurrence. CONCLUSION: No axillary recurrence following SLNB suggests that SLNB may be a good alternative to routine axillary lymph node dissection while providing less surgical morbidity in women with a small breast cancer. However, more patients accumulation and follow-up period will be needed for the final conclusion.

Clinical Results of Coloanal Anastomosis in Radiation-induced Rectovaginal Fistula

PURPOSE: Generally speaking, permanent colostomy has been frequently used as a treatment for radiation-induced rectovaginal fistula. In order to administer an operation accurately, at least two-staged operations have been performed. If anastomosis were to be performed between normal, healthy tissues after removal of irradiation-damaged tissues definite operation could be performed in one stage. We reviewed clinical records to evaluate postoperative recurrence and anal functions in patients who underwent coloanal anastomosis without diverting colostomy as one step due to radiation-induced rectovaginal fistula. METHOD: From Sep. 1994 to Jun. 2000 we did a retrospective study with clinical data of 8 patients who underwent operations due to radiation-induced rectovaginal fistula in Yongdong Severance Hospital. RESULTS: The mean age was 49 years (range 31-61). All patients concurrently received irradiation and induction chemotherapy due to primary, gynecologic malignancies. The total dosage of exposure to radiation was 8,400 cGy in 6 of 8 cases and 8,940 cGy in the remaining 2 cases. TAH with BSO had been undergone before concurrent chemoradiation in 2 cases. The median duration from diagnosis to operation was 29 months (range 16-131) in cases without previous colostomy (n=7) and 7 months in cases with colostom y (n=1). Before the operation, previous surgery had been undergone in 2 cases due to rectovaginal fistula. No recurrences were noted for gynecologic malignancies. The mean distance of fistula opening from anal verge was 3.9 cm (range 2.0-7.0). For 7 out of 8 cases, patients underwent LAR with handsewn coloanal anstomosis and the remaining patient underwent anterior resection. No diverting colostomy was performed for all cases. During the median follow-up period of 25 months (range 7-71), two patients developed anal stenosis. One patient experienced postoperative recurrence for the follow-up period. In terms of sphincter function (n=6) (f/u period>12 months), there were 1 urgency, 1 gas incontinence and 1 night staining. CONCLUSIONS: Although this study is a small scale research in terms of the number of subjects involved, one-staged, handsewn coloanal anastomosis after LAR without colostomy may be proved to be helpful for the patients with radiation-induced rectovaginal fistula. If case selection performed properly, unnecessary operation can be avoided and psychologic resistance can be reduced by this procedure.