Bmi-1 Regulates Snail Expression and Promotes Metastasis Ability in Head and Neck Squamous Cancer-Derived ALDH1 Positive Cells

Recent studies suggest that ALDH1 is a putative marker for HNSCC-derived cancer stem cells. However, the regulation mechanisms that maintain the stemness and metastatic capability of HNSCC-ALDH1+ cells remain unclear. Initially, HNSCC-ALDH1+ cells from HNSCC patient showed cancer stemness properties, and high expression of Bmi1 and Snail. Functionally, tumorigenic properties of HNSCC-ALDH1+ cells could be downregulated by knockdown of Bmi-1. Overexpression of Bmi-1 altered in expression property ALDH1− cells to that of ALDH1+ cells. Furthermore, knockdown of Bmi-1 enhanced the radiosensitivity of radiation-treated HNSCC-ALDH1+ cells. Moreover, overexpression of Bmi-1 in HNSCC-ALDH1− cells increased tumor volume and number of pulmonary metastatic lesions by xenotransplant assay. Importantly, knock-down of Bmi1 in HNSCC-ALDH1+ cells significantly decreased distant metastases in the lungs. Clinically, coexpression of Bmi-1/Snail/ALDH1 predicted the worst prognosis in HNSCC patients. Collectively, our data suggested that Bmi-1 plays a key role in regulating Snail expression and cancer stemness properties of HNSCC-ALDH1+ cells

Network Biology of Tumor Stem-like Cells Identified a Regulatory Role of CBX5 in Lung Cancer

Mounting evidence links cancers possessing stem-like properties with worse prognosis. Network biology with signal processing mechanics was explored here using expression profiles of a panel of tumor stem-like cells (TSLCs). The profiles were compared to their parental tumor cells (PTCs) and the human embryonic stem cells (hESCs), for the identification of gene chromobox homolog 5, CBX5, as a potential target for lung cancer. CBX5 was found to regulate the stem-like properties of lung TSLCs and was predictive of lung cancer prognosis. The investigation was facilitated by finding target genes based on modeling epistatic signaling mechanics via a predictive and scalable network-based survival model. Topologically-weighted measurements of CBX5 were synchronized with those of BIRC5, DNMT1, E2F1, ESR1, MLH1, MSH2, RB1, SMAD1 and TAF5. We validated our findings in another Taiwanese lung cancer cohort, as well as in knockdown experiments using sh-CBX5 RNAi both in vitro and in vivo.National Science Council (China) (NSC grant 100-2325-B-010-010-MY3/98-2314-B-010-024-MY2/97-3111-B075-001-MY3/ 96-2314-075-056-MY3)National Yang-Ming University (Ministry of Education, Aim for the Top University Plan: 96ADD122, 96ADD125, 96ADT191, 97ACD113, 97ACT302, 98ACT302, 98ACD107, 98ACT192 and Brain Research Center-3T-MRI project)))Taipei Veterans General Hospital (98-C1-099/E1-003/ER3-001)Taipei Veterans General Hospital (Joint Projects of VGHUST (98-G6-6/ 98-P1-01/99-P6-39)Chi Mei Medical Center (CMYM9801)Yen-Tjing-Ling Medical Foundation (96/97/98)Taipei City Hospital (96-002-62-092)Technology Development Program for Academia (TDPA; 98-EC-17-A-19-S2-0107)Taiwan. Department of Industrial Technology, Ministry of Economic AffairsNational Science Council (China) (NSC 101-2325-B-010 -009)Taiwan. Department of Health. Cancer Research Center of Excellence (DOH101-TD-C-111-007

Generation of Monoclonal Autoantibodies from Babesia rodhaini-Infected Mice

The presence of anti-erythrocyte autoantibodies in animals infected with various Babesia species is well reported. However, the pathogenesis of autoantibodies in babesiosis is poorly understood. Here, we demonstrated that anti-erythrocyte immunoglobulin (Ig) M and IgG were present in B. rodhaini-infected mice at 6 and 8 days after infection, respectively. Furthermore, we generated monoclonal antibodies against erythrocyte antigen from B. rodhaini-infected mice. Five clones were generated. By Western blotting analysis using whole erythrocyte antigens, one clone reacted with a broad band around 90–150 kDa, and the 2 clones reacted with a band larger than 150 kDa. B. rodhaini-infected mice and/or autoreactive monoclonal antibodies established in this study might be a powerful tool for in vivo pathogenesis studies of autoantibody development in infectious diseases

A Study on Business Method Patent — Explore Current Development of Practice From the Bilski Case

商業方法是否可以獲得專利一直以來有所爭議。早期美國實務在1908年的Hotel Security Checking Co. v. Lorraine Co.一案中，確立了「商業方法除外原則」。然而，在1998年State Street Bank & Trust Co. v. Signature Financial Group一案中，聯邦巡迴上訴法院廢除了「商業方法除外原則」，並且對專利適格性採用了寬鬆的「有用、具體與有形的結果」判斷標準，從此打開的商業方法專利的大門，但也導致商業方法專利申請趨於氾濫之情形，而學說界或實務界，對於商業方法能否取得專利爭議一直未曾停歇。直到2008年Bilski一案中，聯邦巡迴上訴法院判決結果雖未否認商業方法之專利適格性，但轉而採取較為嚴格的「機器或轉換測試」判斷標準。不過，最高法院並未完全採納聯邦巡迴上訴法院的見解，所以在Bilski案之後，實務對於商業方法適格性的判斷仍未趨一致。直到2014年CLS案中，最高法院對於商業方法專利適格的判斷提供了較為明確的判斷方式。 相對於美國仍陷於專利適格性之爭論，日本及歐洲實務界對於商業方法專利除了探討專利適格性即可專利性要件之外，亦著重在進步性要件之判斷。日本實務對於可專利性採取「整合硬體資源」判斷標準，歐洲實務則強調「技術性」，兩者對於可專利性均設下較寬鬆之門檻，但在進步性的審查則採取嚴格的標準，並且著重在「技術貢獻」的判斷。本研究認為，我國實務雖然受到美國實務的影響，但目前日本及歐洲在商業方法專利的審查較為明確，因此我國實務不妨對可專利性採取較寬鬆的門檻，但對於進步性採取較嚴格的審查標準。Whether business methods can be patented has been somewhat controversial. Early in 1908 in the U.S. practice, the Hotel Security Checking Co. V. Lorraine Co.''s Case established "business methods exception principle." However, in the State Street Bank & Trust Co. V. Signature Financial Group case in 1998, the Federal Circuit Court of Appeals repealed of this principle and adopted a liberal "useful, concrete and tangible result" criterion. Since then the number of business method patent applications increased significantly. However, business method patent dispute had not been able to stop. Until the Bilski case in 2008, although the Federal Circuit Court of Appeals did not deny that business method patents are patentable, it took a more stringent “machine or transformation test” criterion. Nevertheless, the Supreme Court did not fully adopt the Federal Circuit Court of Appeals opinion. After the Bilski case, practice in U.S. has not yet reached a consistent interpretation about the patentability of the business method patent. Until the CLS case in 2014, the Supreme Court provided a more definitive judgment to determine the patentability of the business method patent. Contrast to the controversy of patentability in the U.S., the practice in Japan and Europe discussed not only the patentability but also the inventive step (non-obviousness). Japanese practice take “integrated hardware resources” criterion for patentability, and European practice emphasizes “technical character.” Both of them set the lower threshold for the patentability, but they also adopted rigorous review for inventive step. The study suggests that despite the impact of the U.S. practice, the current standard of Japan and Europe practice is more clearly, so the Taiwan practice may take the similar standard of Japan and Europe.目錄 誌謝 I 中文摘要 II 英文摘要 III 第一章 緒論 1 第一節 研究動機 1 第二節 研究目的 2 第三節 論文架構 2 第二章 商業方法專利概論 4 第一節 商業方法的定義 4 第二節 商業方法專利的分類 5 第一項 IPC國際專利分類 5 第二項 美國專利分類 6 第三項 日本專利分類 7 第四項 我國專利分類 7 第三節 商業方法專利與軟體專利之關係 7 第三章 美國商業方法專利之發展 9 第一節 商業方法專利要件規定 9 第二節 專利適格性要件 10 第一項 Bilski案前商業方法專利發展 10 第二項 Bilski v. Kappos案 22 第三項 Bilski案後相關案例探討 38 第三節 非顯而易知性要件 58 第一項 非顯而易知性之審查 58 第二項 實務相關案例探討 62 第四節 商業方法專利申請核准概況 66 第五節 本章小結 67 第四章 日本商業方法專利之發展 69 第一節 相關法規及審查基準 69 第一項 可專利性要件 69 第二項 進步性要件 71 第二節 不該當商業方法專利之事例 76 第一項 不該當於專利法的發明 76 第二項 不具有進步性 78 第三節 實務相關案例探討 86 第一項 知的財產高等裁判所平成17年（行ケ）10698號判決 86 第二項 知的財產高等裁判所平成20年（行ケ）10151號判決 87 第三項 知的財產高等裁判所平成17年（行ケ）10335號判決 91 第四項 知的財產高等裁判所平成15年（行ケ）240號判決 92 第五項 知的財產高等裁判所平成17年（行ケ）10084號判決 94 第六項 知的財產高等裁判所平成19年（行ケ）10327號判決 96 第四節 商業方法專利申請及核准概況 99 第五節 本章小結 101 第五章 歐洲商業方法專利之發展 102 第一節 相關法規及審查基準 102 第一項 可專利性要件 102 第二項 進步性要件 105 第二節 實務相關案例探討 109 第一項 1994年Sohei案 109 第二項 2000年Pension Benefit System（PBS）案 112 第三項 2003年RICOH案 115 第四項 2004年Hitachi案 118 第三節 本章小結 122 第六章 我國商業方法專利之發展 123 第一節 相關法規及審查基準 123 第一項 可專利性要件 123 第二項 進步性要件 129 第二節 實務相關案例探討 134 第一項 智慧財產法院101年度民專上更(二)字第5號民事判決 134 第二項 智慧財產法院102年度民專上字第25號民事判決 138 第三項 智慧財產法院102年度行專訴字第61號行政判決 144 第四項 智慧財產法院102年度行專訴字第80號行政判決 150 第五項 智慧財產法院102年度行專訴字第88號行政判決 155 第三節 商業方法專利申請核准概況及近期核准專利評析 162 第一項 商業方法專利申請核准概況 162 第二項 近期商業方法專利核准之評析 163 第四節 本章小結 165 第七章 結論 167 第一節 各國商業方法專利要件之比較 167 第二節 對我國實務發展之建議 169 參考文獻 17

Allozyme Variation of Populations of Castanopsis carlesii (Fagaceae) Revealing the Diversity Centres and Areas of the Greatest Divergence in Taiwan

• Background and Aims The genetic variation and divergence estimated by allozyme analysis were used to reveal the evolutionary history of Castanopsis carlesii in Taiwan. Two major questions were discussed concerning evolutionary issues: where are the diversity centres, and where are the most genetically divergent sites in Taiwan

Age-Specificity of Toxoplasma gondii Seroprevalence in Sheep, Goats and Cattle on Subsistence Farms in Bangladesh

Toxoplasma gondii is a zoonotic protozoan parasite that infects humans and domestic animals. In this study, the seroprevalence of T. gondii antibodies was investigated using serum samples collected from 83 sheep, 146 goats and 37 cattle from a dozen subsistence farms in Bangladesh. Fifty-eight out of 83 sheep (69.9%), 89 out of 146 goats (61.0%) and 10 out of 37 cattle (27.0%) were seropositive for the parasite. Seroprevalence in young goats (<1 year old) was significantly lower than that of the adult goats (>1 year old). In contrast, seroprevalence for young and adult sheep was similar. These results indicate that acquired infection with T. gondii occurs in this region of Bangladesh, at least among goats

Photothermolysis of glioblastoma stem-like cells targeted by carbon nanotubes conjugated with CD133 monoclonal antibody

CD133+ cells in glioblastoma (GBM) display cancer stem cell-like properties and have been considered as the culprit of tumor recurrence, justifying exploration of potential therapeutic modalities targeting CD133+ cancer stem-like cells (CSCs). For photothermolysis studies, GBM-CD133+ and GBM-CD133- cells mixed with various ratios were challenged with single-walled carbon nanotubes (SWNTs) conjugated with CD133 monoclonal antibody (anti-CD133) and then irradiated with near-infrared laser light. Results show that GBM-CD133+ cells were selectively targeted and eradicated, whereas GBM-CD133- cells remained viable. In addition, in vitro tumorigenic and self-renewal capability of GBM-CD133+ treated with localized hyperthermia was significantly blocked. Furthermore, GBM-CD133+ cells pretreated with anti-CD133-SWNTs and irradiated by near-infrared laser 2 days after xenotransplantation in nude mice did not exhibit sustainability of CSC features for tumor growth. Taken altogether, our studies demonstrated that anti-CD133-SWNTs have the potential to be utilized as a thermal-coupling agent to effectively target and destroy GBM CSCs in vitro and in vivo. From the Clinical Editor: Glioblastoma remains one of the most notorious cancer from the standpoint of recurrence and overall resistance to therapy. CD133+ stem cells occur among GBM cells, and may be responsible for the huge recurrence risk. This paper discusses a targeted elimination method of these cells, which may enable more efficient therapy in an effort to minimize or prevent recurrence. © 2011 Elsevier Inc