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A polysaccharide (ESPS) purified from Eupolyphaga sinensis Walker by ion exchange chromatography and gel chromatography was investigated, including its structure characterization and antitumor activity. The results showed that ESPS was composed of rhamnose, fucose, arabinose, xylose, glucose, and galactose in a molar ratio of 7.4: 3.1: 13.9: 9.3: 39.7: 26.5, with the mean weight (Mw) of 2.14 × 104 Da; the main chain of ESPS was mainly composed of → 4) - α - D - Glcp - (1 → and → 3) - β - D - Galp - (1 →, and the side chains were connected to the main chain through the O-6 atom of glucose and O-4 and O-6 atom of galactose. In addition, ESPS promoted the lymphocyte proliferation and inhibited liver cancer cells growth through enhancing lymphocyte activity in vitro, mainly NK cells. Moreover, ESPS markedly stimulated immunity in H22-bearing mice by increasing the spleen and thymus indices and effectively inhibited H22 cell growth in vivo. These data indicated that ESPS was a polysaccharide component possessing high anti-hepatocellular carcinoma activity, representing a potential immunotherapy candidate for the treatment of liver cancer.

Tubiechong (Eupolyphaga sinensis) is an important material used in traditional Chinese medicine (TCM). However, the immunoregulation effects of E. sinensis Lyophilized Powder (ESL) are uncl...

Tubiechong, in Chinese medicine, denotes the dried female insects of Eupolyphaga sinensis Walker (ESW) or Polyphaga plancyi Bolivar (PPB). As a traditional insect-type, in medicine, it has been historically utilized to treat bruises, fractures, amenorrhea, postpartum blood stasis, lumps and relieving pain. We herein have performed a systematic survey involving the chemical and biological studies in the past decades to reveal the value of such insect resources for their development and clinical utilization. Chemical studies indicated that Tubiechong generated many active compounds, including proteins, amino acids, peptides, fatty acids, alkaloids, nucleosides, polysaccharides, fat-soluble vitamins and mineral elements. Tubiechong or its extract has a wide range of activities including anticoagulation and anti-thrombosis, anti-tumor, antioxidant, immune regulation, blood lipid regulation and hepatoprotection. Finally, a periodic mini-review was conducted to summarize such chemical and pharmacological profiles of Tubiechong medicine. The active peptides in Tubiechong are majorly focused in this review and introduced as one important aspect since there is much literature and huge investigative interest in it. Traditional medical use of the insect was also stressed in this review associating with its disease-eliminating actions by promoting blood circulation or eliminating tissue-swelling pains, which might play important roles in anticancer practices or investigation. In accordance with the modern pharmacological progress, Tubiechong and its extracts indeed exerted antitumor actions through multiple pathways, such as interfering with tumor biological behaviors (growth, apoptosis, invasion, metastasis and angiogenesis), and regulating host immune function. To some extent, this knowledge would provide a basis for further research and application of Tubiechong medicine.

sinensis Saussure, 1869: 282-283 [Homoeogamia] Nord de la Chine; Pekin. Two ♂. The MHNG collection contains one ♂ specimen (MHNG-ARTO-20468) with labels: “ ♂ Pekin, Chine, M. H de Saussure ” [handwritten on ruled white card]; “ Heterogamia sinensis Sauss. ♂ ” [handwritten on yellow paper]; Heterog. sinensis Sauss. ” [handwritten on blue paper]; “ Syntypus ” [printed on red paper]. The specimen is directly pinned with the left wings spread and the right wings folded. There is insect feeding damage to the head, and the left antenna, the last tarsal segment of the right middle leg, the last tarsal segment of the left hind leg and all of the tarsi of the right hind leg are missing. Box Blattodea 59. The whereabouts of the other syntype is unknown. This species is not listed in the CSF database. Kirby (1903: 379) considered Polyphaga sinensis (Saussure, 1869) to be a junior homonym of P. sinensis Walker, 1868 and proposed the replacement name P. limbata. Replaced by Eupolyphaga limbata (Kirby, 1903), Published as part of Hollier, John, Marshall, Judith & Legendre, Frédéric, 2020, An annotated list of the Blattodea (Insecta) described by Henri de Saussure. Part 1: the Corydiidae, pp. 341-348 in Revue suisse de Zoologie 127 (2) on page 347, DOI: 10.35929/RSZ.0023, http://zenodo.org/record/5743830, {"references":["Saussure H. de 1869. Melanges Orthopterologiques Fascicule II. Blattides et Phasmides. Memoires de la Societe de Physique et d'histoire naturelle de Geneve 20: 227 - 328.","Kirby W. F. 1903. Notes on Blattidae etc., with descriptions of new genera and species in the collection of the British Museum, South Kensington. No. III. The Annals and Magazine of Natural History 12: 373 - 381.","Walker F. 1868. Catalogue of the specimen of Blattariae in the collection of the British Museum. British Museum, London 239 pp."]}

Huoxuezhitong capsule (HXZT, activating blood circulation and relieving pain capsule), has been applied for osteoarthritis since 1974. It consists of Angelica sinensis (Oliv.) Diels, Panax notoginseng (Burkill) F. H. Chen ex C. H., Boswellia sacra, Borneol, Eupolyphaga sinensis Walker, Pyritum. However, the direct effects of HXZT on osteoarthritis and the underlying mechanisms were poorly understood. In this study, we aimed to explore the analgesia effect of HXZT on MIA-induced osteoarthritis rat and the underlying mechanisms. The analgesia and anti-inflammatory effect of HXZT on osteoarthritis in vivo were tested by the arthritis model rats induced by monosodium iodoacetate (MIA).. Mechanistic studies confirmed that HXZT could inhibit the activation of NF-κB and down-regulate the mRNA expression of related inflammatory factors in LPS-induced RAW264.7 and ATDC5 cells. Furtherly, in LPS-induced RAW264.7 cells, HXZT could suppress NF-κB via inhibiting PI3K/Akt pathway. Taken together, HXZT capsule could ameliorate MIA-induced osteoarthritis of rats through suppressing PI3K/ Akt/ NF-κB pathway.

The traditional medicine-Eupolyphaga sinensis walker is a rich source of functional proteins and peptides. In this study, Eupolyphaga sinensis walker were identified to include fifteen amino acids, of which hydrophobic amino acids accounted for 46.7%. Eupolyphaga sinensis walker polypeptides (EPs) were extracted by sequentially hydrolyzing with pepsin and trypsin. EPs could effectively scavenge 1,1-diphenyl-2-picrylhydrazyl radical (DPPH·), superoxide anion radical (O2−·), and hydroxyl radical (OH-·), and reduce ferric solution in vitro. EPs also enhanced the activities of Ssuperoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPH-Px), increased the contents of hydroxyproline (HYP), and reduced the content of Malondialdehyde (MDA) in photoaged mice skin. Histological study confirmed that EPs improved UV irradiation-induced damage of skin texture and morphology. Therefore, the extracted EPs are effective antioxidants and can serve as powerful treatment for skin photoaging.

Eupolyphaga sinensis (Walker, 1868) (Figs. 4 A–K; 13 A–C; 17 A–D; 38 A–B, L; 39 A–C) Polyphaga sinensis Walker, 1868: 14, female holotype, “North China ”; Chopard 1922: 194, “ Chine. septentrionale”. Homoeogamia sinensis Saussure, 1869: 282, 2 males, “Le Nord de la Chine, Pékin ”. Syn. nov. Heterogamia sinensis: Dohrn 1888: 132; Saussure 1893: 311. Heterogamia dohrniana Saussure, 1893: 309, “Sina septentrionalis”. Polyphaga limbata Kirby, 1903: 379, new name for Homoeogamia sinensis Saussure; Kirby 1904: 172, “N. China ”; Chopard 1922: 194, “ Chine: Pékin; montagnes au nord de Pékin ”; Chopard 1929: 269, Discussion on P. limbata. Eupolyphaga sinensis: Chopard 1929: 262; Wu 1935: 29; Bey-Bienko 1950: 285; Princis 1962: 53; Woo 1987: 433; Feng, Guo & Woo 1997: 35. Eupolyphaga limbata: Wu 1935: 29, “N. China ”; Princis 1959: 149, discussion on Eupolyphaga limbata, treated E. limbata as a valid species and synonymized E. thibetana under E. limbata; Princis 1962: 54. Material examined. CHINA: Beijing: 1 male and 2 females (SWU, all preserved in 100% alcohol), Xishan Forest Park [西山森林公园], Haidian District [海淀区], 28.IV.2015, Bing-Qiang Wang collected and reared; 7 males (SWU), Haidian District, VII.2006, Tao-Li Zhang, Su-Ying Wu & Shu-Hua Qiao leg.; 1 male (SWU), Haidian District, 6.IX.1964, no collector recorded; 4 males (SWU), Haidian District, V-VII.1961, no collector recorded; 3 males (SWU), Jinshan [金山], 7,14,19. VIII.1984, Guo-Wei Wang leg.; 1 male (BJMNH), Jinshan, 17.IX.1983, no collector recorded; 3 males (SWU), Songshan Natural Reserve [松山自然保护区], Yanqing County [延庆县], 13.VII.2013, Yi-Zhou Liu leg.; 5 males (IZCAS), Mt. Baihuashan [百花山], Mentougou District [门头沟区], 27.VIII.1973, Yin-Heng Han leg.; 1 male (BJMNH), Xiaolongmen Forest Park [小龙门森林公园], Mentougou District, 24.VIII.1979, Si-Kong Liu leg.; 4 males (IZCAS), Xiaolongmen Forest Park, Mentougou District, 1100m, 20.VI.1991, Pei-Yu Yu leg.; 1 male (IZCAS), Nancun Village [南村], Tanzhesi Town [潭柘寺镇], Mentougou District, 6.VII.1972, no collector recorded; 1 male (IZCAS), Baiyunxia [白云峡], Miyun County [密云县], 20.VII.2009, Zhe Wang & Xing-Yi Li leg.; 1 male (IZCAS), Xijiao Park [西郊公园], Beijing City, 17.VI.1954, Guang Gao leg.; 1 male (IZCAS), Pinggu District [平谷区], 14.VI.1975, no collector recorded; 1 male (IZCAS), Pinggu District, 29.VI.1975, Yong-Shan Shi leg.; 1 male (IZCAS), Shahe [沙河], Changping District [昌平区], VI.1959, no collector recorded; 2 males (BJMNH), Jiuxianqiao [酒仙桥], Zhaoyang District [朝阳区], 22, 29.VIII.2001, no collector recorded; 1 male (SWU), Tiantan / The Temple of Heaven [天坛], Dongcheng District [东城区], 25.VII.1973, Si-Kong Liu leg.; 3 males (IZCAS), Beijing City, 1.IX.1972, no collector recorded; 1 male (IZCAS), “ University of Agricuture ”, 1948, no collector recorded; 1 male (SWU), Beijing Agricultural University, Beijing City, 20. IX.1957, Ji-Kun Yang leg.; 1 male (SWU), Peiping (now Beijing City), 30.VI.1947, no collector recorded; 1 nymph (SWU), Xiaolongmen Forest Park, Mentougou District, 18.VIII.2014, Yi-Zhou Liu leg.; several nymphs (BJMNH), Taoranting Park [陶然亭公园], Xicheng District [西城区], VII-IX.2003, Feng Yuan and Hong- Zheng Yang leg.; 1 nymph (SWU), Beijing, VIII.1977, Fo-Ching Woo leg.; 4 nymphs (SWU, in 100% alcohol), Beijing Botanical Garden [北京植物园], 27.VII.2016, Yi-Zhou Liu leg. Hebei: 2 males (MHBU), Guanting tree farm [官厅林场], Huailai County [怀来县], Zhangjiakou City [张家口市], 15.VIII.1999, Ai-Min Shi leg.; 1 female and 3 nymphs (MHBU), Guanting Town [官厅镇], Huailai County, Zhangjiakou City, 15.VII.1999, Ai-Min Shi leg.; 1 nymph (MHBU), Yongding River [永定河], Huailai County, Zhangjiakou City, 15.VII.2000, Xiu-Juan Yang leg.; 2 males (MHBU), Yangjiaping tree farm [杨家坪林场], Zhuolu County [涿鹿县], Zhangjiakou City, 11- 19.VII.2002, Ai-Min Shi leg.; 1 male (MHBU), Yangjiaping [杨家坪], Zhuolu County, Zhangjiakou City, 4- 12.VII.2004, students of Hebei University leg.; 2 males (MHBU), Yangjiaping, Zhuolu County, Zhangjiakou City, 14-24.VII.2005, Guo-Dong Ren and Hui-Ran Hua leg.; 1 male (MHBU), Yangjiaping, Zhuolu County, Zhangjiakou City, 10-24.VII.2005, Jing Li and Hui-Ran Hua leg.; 2 females and 6 nymphs (MHBU), Yangjiaping, Zhuolu County, Zhangjiakou City, 4-12.VII.2004, students of Hebei University leg.; 2 females and 5 nymphs (MHBU), Yangjiaping tree farm, Zhuolu County, Zhangjiakou City, 11-19.VII.2002, Ai-Min Shi et al. leg.; 1 nymphs (MHBU), Sangyuanjian tree farm [桑园涧林场], Zhuolu County, Zhangjiakou City, 28.VII.1999, Wen- Min Cui leg.; 2 males (SWU), “Chahar, Yangkiaping” (now Yangjiaping, Zhangjiakou City), 29-30.VII.1937, O. Piel leg.; 1 male (IZCAS), “ Chahar, Yangkiaping ”, 30.VII.1937, O. Piel leg.; 1 male (MHBU), Shanjiankou [山涧 口], Xiaowutaishan Mountain [小五台山], Weixian County [蔚县], Zhangjiakou City, 18-21.VII.2005, Jing Li, Feng-Yan Wang leg.; 4 males (MHBU), Jinhekou [金河口], Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 29.VII.1998, Shi-Yu Liu leg.; 2 males (MHBU), Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 11-17.VII.2002, Students of Hebei University leg.; 2 males (MHBU), Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 10.VII.2001, students of Hebei University leg.; 1 male, 1 female and 7 nymphs (MHBU), Jinhekou, Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 18-19.VI.2009, Xin-Pu Wang, Zhen-Hua Gao leg.; 1 female and 9 nymphs (MHBU), Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 5-8.VII.2001, students of Hebei University leg.; 3 females and 18 nymphs (MHBU), Jinhekou, Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 7-11.VII.2006, students of Hebei University leg.; 1 nymph (MHBU), Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 27-30.VIII.2007, Ya-Lin Li leg.; 6 nymphs (MHBU), Jinhekou, Xiaowutaishan Mountain, Weixian County, Zhangjiakou City, 10-11.VII.2005, Jing Li and Feng-Yan Wang leg.; 9 males (IZCAS), Yangjiaping, Xiaowutaishan Natural Reserve, 820m, 2-4.VIII.2011, Jian Yao leg.; 5 males (IZCAS), Yangjiaping, Xiaowutaishan Natural Reserve, 880m, 29-30.VII.2009, Jian Yao leg.; 1 male (IZCAS), Xiaowutaishan Natural Reserve, 1200m, 10.VIII.1964, Bing-Qian Li leg.; 1 male (IZCAS), Xiaowutaishan Natural Reserve, 1200m, 25.VIII.1964, Chun-Xing Wang leg.; 1 male (IZCAS), Xiaowutaishan Natural Reserve, 1200m, 20.VIII.1964, Yin-Heng Han leg.; 2 males (MHBU), Mt. Guangtoushan [光头山], Pingquan County [平泉县], Chengde City [承德市], 2.VIII.1998, Zhe Li et al. leg.; 2 males (MHBU), Mt. Guangtoushan, Pingquan County, Chengde City, 21.VII-6.VIII.1998, Zhe Li et al. leg.; 3 males (MHBU), Mt. Guangtoushan, Pingquan County, Chengde City, 21.VII-6.VIII.1998, Chao Zhang leg.; 3 males (IZCAS), Zhaoxian County [赵县], Shijiazhuang City [石家庄市], 2.VII.2009, no collector recorded; 1 male (MHBU), Nansizhang Village [南寺掌村], Jingxing County [井陉县], Shijiazhuang City, 28-29.VII.1998, Li-Xin Gao leg.; 2 males (MHBU), Nansizhang Village, Jingxing County, Shijiazhuang City, 28-29.VII.1998, Zhi-Qiang Pu leg.; 2 males (MHBU), Gaoyu Village [高峪村], Jingxing County, Shijiazhuang City, 22-28.VII.1998, Xin-Sheng Tian et al. leg.; 1 male (MHBU), Xinzhuang tree farm [辛庄林场], Jingxing County, Shijiazhuang City, 30.VIII.1998, Ming Bai leg.; 1 male (MHBU), Suobao tree farm [索堡林场], Shexian County [涉县], Handan City [邯郸市], 21.VII.1998, Li-Xin Gao leg.; 3 males and 1 nymph (MHBU), Guantao County [馆陶县], Handan City, 18.VII.2000, Zhi-Jian Wang leg.; 1 male (MHBU), Longquanguan [龙泉关], Fuping County [阜平县], Baoding City [保定市], 3- 5.VIII.1998, Ai-Min Shi et al. leg.; 7 males (MHBU), Mt. Baishishan [白石山], Laiyuan County [涞源县], Baoding City, 1-3.VIII.1999, Ai-Min Shi et al. leg.; 2 nymphs (MHBU), Mt. Baishishan, Laiyuan County, Baoding City, 11.VII.1999, Ai-Min Shi and Xiu-Juan Yang leg.; 1 male (MHBU), Mt. Baishishan, Laiyuan County, Baoding City, 22.VIII.1999, Xin-Jiang Li leg.; 2 males (MHBU), Shipuxia Gorge [十瀑峡], Laiyuan County, Baoding City, 4.VIII.1999, Ai-Min Shi et al. leg.; 1 nymph (MHBU), Hekou Township [河口乡], Shunping County, [顺平县] Baoding City, 5.IV.2004, Guo-Dong Ren leg.; 3 males (MHBU), Mt. Zushan [祖山], Qinhuangdao County [秦皇 岛市], 14.VII.1998, Xin-Jiang Li et al. leg.; 17 nymphs (MHBU), Forest Park of Xingtai City [邢台市森林公园], 18.VII.1998, Wei Wang et al. leg.; 2 females (MHBU), Mt. Mangshan [蟒山], Qian’an City [迁安市], Tangshan City [唐山市], 28.VII.2000, Ai-Min Shi leg. Henan: 1 male (SWU), Xinxiang City [新乡市], 29.VI.1974, no collector recorded; 1 male (SWU), Xinxiang City, 8.VII.1960, no collector recorded; 1 male (SWU), Xinxiang City, 25.VIII.1974, no collector recorded; 1 male (SWU), Xinxiang City, 8.IX.1974, no collector recorded; 2 males (SWU), Xinxiang City, 27.VI.1973, no collector recorded; 1 male (SWU), Xinxiang City, 27.VIII.1972, no collector recorded; 1 female and 1 nymph (MHBU), Jinglonggong Township [荆隆宫乡], Fengqiu County [封丘 县], Xinxiang City, 17.VII.2007, Hao-Yu Liu and Zhen-Hua Gao leg.; 1 male (MHBU), Baligou [八里沟], Shangbali Town [上八里镇], Huixian City [辉县], Xinxiang City, 12.VII.2002, Xiu-Juan Yang leg.; 2 nymphs (MHBU), Shangbali Town, Huixian City, Xinxiang City, 27.VII.2006, Feng-Yan Wang and Wen-Jing Huang leg.; 1 male (MHBU), Tongbai County [桐柏县], Nanyang City [南阳市], 600-800m, 16.VII.2001, Zhe Li and Ming Bai leg.; 1 male (MHBU), Tongbaishan Mountain [桐柏山], Tongbai County, Nanyang City, 25.VII.2010, Yi-Bin Ba and Zhen-Xing Zhang leg.; 2 males (MHBU), Baotianman [宝天曼], Neixiang County [内乡县], Nanyang City, 20.VIII.2008, Guo-Dong Ren and Qi-Qi Wu leg.; 1 female and 6 nymphs (MHBU), Huangshi’an [黄石庵], Xixia County [西峡县], Nanyang City, 18.VIII.2008, Guo-Dong Ren and Qi-Qi Wu leg.; 1 male (IZCAS), Xindian [辛店], Xinyang City [信阳市], 27.VIII.1969, Ding-Gang Hu leg. Inner Mongolia / Neimenggu: 5 males (IZCAS), Houyikenggou [后一坑沟], Mt. Helanshan [贺兰山], Alxa Left Banner [阿拉善左旗], 1954m, 10.VIII.2010, Mei- Ying Lin leg.; 1 male (IZCAS), Darhan Muminggan Joint Banner [达尔罕茂明安联合旗], Baotou City [包头市], 7.VIII.1971, no collector recored; 2 nymphs (MHBU), Bayan Obo [白云鄂博], Baotou City, 3.VIII.1991, Guo- Dong Ren leg.; 2 males, 3 females, 14 nymphs (MHBU), Hangali Lake [翰嘎利湖], Horqin Right Wing Middle Banner [科尔沁右翼中旗], 310m, 29.VII.2012, Yi-Bin Ba and Yi-Ping Niu leg.; 4 nymphs (MHBU), Daban Town [大板镇], Bairin Right Banner [巴林右旗], Chifeng City [赤峰市], 19.VI.1994, Guo-Dong Ren leg.; 1 nymph (MHBU), Chifeng City, 27.VI.1994, Guo-Dong Ren leg.; 1 nymph (MHBU), Shangwan Town [上湾镇], Ejin Horo Banner [伊金霍洛旗], Ordos City [鄂尔多斯市], 18.VII.2012, Yu Zhao and Ji-Bin Liang leg.; 1 female (IZCAS), Xiaochengzi Township [小城子乡], Jalaid Banner [扎赉特旗], 21.VI.1970, Zhi-Zi Chen leg. Liaoning: 1 female (MHBU), Haigang District [海港区], Dalian City [大连市], 31.VII.2003, Xiu-Juan Yang and Shan-Shan Liu leg.; 1 nymph (SWU, in 100% alcohol), Xishan Reservoir [西山水库], Dalian City, 142m, 1.VII.2016, Lu-Yu Wang leg.; 3 nymphs (SWU, in 100% alcohol), Tongnai Village [同乃村], Fuxin County [阜新县], 329m, 31.VII.2016, Lu-Yu Wang leg.; 2 males (SWU), Shicheng Island [石城岛], Zhuanghe City [庄河市], Dalian City, 22.VIII.2016, Ye-Jie Lin leg. Jilin: 1 male (IZCAS), Manjiang Town [漫江镇], Fusong County [抚松县], 21.IV.1955, no collector recoreded. Tianjin: 1 female (IZCAS), “ Jixian County [蓟县], Heibei Province (now belongs to Tianjin)”, 21.IX.1954, no collector recorded. Shanxi: 1 male (SWU), Wangjiaying Village, Yuncheng City, VIII.1975, no collector recorded; 4 nymphs (MHBU), Lanxian County [岚县], Lvliang City [吕梁市], 10.V.1995, no collector recorded; 1 nymph (MHBU), Lishi District [离石区], Lvliang City, 9.V.1995, no collector recorded. Shaanxi: 6 females, 3 nymphs (MHBU), Yanhe River [延河], Yan’an City [延安市], 6.V.1995, no collector recorded; 2 nymphs (MHBU), Huangling County [黄陵县], Yan’an City, 17.VII.1992, Guo-Dong Ren leg. Ningxia: 14 males and 3 nymphs (MHBU), Mt. Xumishan [须弥山], Guyuan City [固原市], 17.VII.2009, Xin-Pu Wang & Hong-Fan Ran leg.; 2 malesand 1 female (MHBU), Dachong Temple [大冲寺], Haiyuan County [海源县], Zhongwei City [中卫市], 25.VIII.1986, Guo-Dong Ren leg.; 3 males (MHBU), Pingluo County [平罗县], Shizuishan City [石嘴山市], 30.VI.1989. Shandong: 2 males (IZCAS), Laoshan tree farm [崂山林场], Qingdao City [青岛市], 26.IX.1957, Yin-Heng Han leg.; 1 male (IZCAS), Jinan City [济南市], 12.VII.1973, no collector recorded; 2 males (IZCAS), Qihe County [齐河县], Dezhou City [德州市], 12.VII.1969, Yong-Lin Chen leg.; 2 females (MHBU), Aishan Forest Park [艾山森林公园], Penglai City [蓬莱市], 7-8.V.2007, Feng-Yan Wang, Ji- Niang Wang & Qi-Qi Wu leg.; 1 female and 3 nymphs (MHBU), Mt. Kunyushan [昆嵛山], Muping District [牟平 区], Yantai City [烟台市], 9-11.V.2007, Feng-Yan Wang, Ji-Niang Wang & Qi-Qi Wu leg.; 6 nymphs (MHBU), Mt. Dazeshan [大泽山], Pingdu City [平度市], 5-6.V.2007, Feng-Yan Wang, Ji-Niang Wang & Qi-Qi Wu leg.; 1 female and 1 nymph (MHBU), Dawa tree farm [大洼林场], Pingyi County [平邑县], Linyi City [临沂市], 13- 15.V.2007, Feng-Yan Wang, Ji-Niang Wang & Qi-Qi Wu leg. Jiangsu: 1 female (SWU), Wujing District, VII.1975, no collector recorded. Shanghai: 1 female (SWU), 1940, labelled “CHINE, Prov. KAINGSU Shanghai ”, handwriting: “ Eupolyphaga sinensis (Wlk) ” and “1940 Det. E.R. Thinkham ”. Anhui: 1 male (SWU), “ Suxian County (now Suzhou City) [宿州市]”, 30.VIII.1973, no collector recorded; 2 males (IZCAS), Wuyang Village [吴阳村], Huoqiu County [霍邱县], Lu’an City [六安市], 11.VI.1966, no collector recorded. Chongqing: 1 male (SWU), Wulong County [武隆县], 7.VII.1989, Long-Long Yang leg. Guizhou: 1 male (SWU), Meitan County [湄潭县], Zunyi City [遵义市], VI.1974, no collector recorded. Yunnan: 1 male (SWU), Yuanjiang [元江 县], 30.VI.1986, Ping-Zhang Feng leg.; 4 males (NWAFU), Menglun Town [勐仑镇], Mengla County [勐腊县], Xishuangbanna Prefecture [西双版纳], 19.V.1991, Ying-Lun Wang and Run-Gang Tian leg. Type material examined. Holotype of Polyphaga sinensis (NHM, No. #876173), female, CHINA: red bordered round label: “ Holo-type ”; round label: “ North China ”; “Type.”; “ sinensis.”; “ Polyphaga sinensis Walker ”. Syntypus [Syntype] of Homoeogamia sinensis (GMNH), 1 male, CHINA: Beijing: red label: “ Syntypus ”; “male Pekin, Chine./ M H{unreadable}”; blue label: “ Heterog. sinens ”; yellow label: “ Heterogamia sinensis, Sauss. ”. Diagnosis. The male E. sinensis superficially resembles E. shennongensis sp. nov., but can be easily distinguished from the latter by the following characters: 1) legs and abdomen pale yellowish (Fig. 4 B), while legs and abdomen brown in E. shennongensis sp. nov. (Fig. 5 D); 2) the yellowish white margin in pronotum usually wider (Fig. 13 A–B), while narrower in E. shennongensis sp. nov. (Fig. 13 D–E); 3) styli small (Fig. 17 B), while larger in E. shennongensis sp. nov. (Fig. 18 B). This species is also historically confused with E. thibetana and E. yunnanensis in male, but it can be distinguished from the latter two by the dense and interfered maculae on tegmina (Fig. 4 A, C, I), while the maculae in E. thibetana and E. yunnanensis are isolated and much more scattered (Fig. 6 A, F, H, J, L). Redescription. Male. General: measurements (mm): body length: 17.7–23.4, overall length: 28.5–33.5, pronotum length×width: 4.7–6.2×8.1–10.2, tegmen length: 25.8–30.1. Size median to large, pale yellow, with brown maculae on tegmina (Fig. 4 A–C). Head: longer than width. Vertex black. Interocular space moderate, narrower than the distance between ocelli. Ocelli large, ocelli ridge distinct, curved, pubescent. Frons wrinkled, dark brown, lateral with two dimples which next to the antennal sockets. Antennae brown. Clypeus distinct, anteclypeus yellowish white, post-clypeus yellowish brown to brown, median with a longitudinal line divides it into two parts, sometimes the two parts are occupied by two large brown spots. Labrum small, yellowish brown, distal margin white, depressed. Maxillary palpi and labial palpi brown, with joint parts yellow (Fig. 13 B–C). Pronotum: dark brown, anterior margin widely whitish yellow, subtransparent. Surface generally with small yellowish pubescence, anterior and lateral margins with additional long brown setae. Apex of anterior margin slightly convex and truncated; lateral fore margins oblique; lateral parts straight, forming an angle with the anterior margin; hind margin slightly curved (Fig. 13 A). Tegmina and wings: exceeded the end of abdomen about 10.4–14.5mm. Tegmina light brownish yellow, hyaline, densely with brown speckles, the speckles mix together, not isolated, divide the hyaline part into many small irregular spots, the brown and hyaline speckles usually evenly distributed. Wings brownish hyaline, slightly maculated, venation distinct. Legs: whitish yellow, covering with brown setae, tibial spines reddish brown. Abdomen: pale whitish yellow. Supra-anal plate in ventral view well pubescent, apex protruded; two median sclerites elongate; cerci short (Fig. 17 A). Subgenital plate unicolored for the exposed part, hind margin setose, nearly straight; styli small, asymmetrical, left longer than the right (Fig. 17 B). Genitalia: well sclerotized. Left phallomere: anterior of L1 round, with a large beak-shaped process towards left, two hind lobes stout; L2 thick, roundly curved, the right end widened; L3 robust, short, curved; pda well developed, paa gentle protruded; L8 large. Right phallomere: small. R1M hollowed; R1L well sclerotized, thick; R3 concave; R2 generally divided into two chunks, the outer one much larger than the inner one (Fig. 17 C–D). Female. Measurements (mm): body length: 23.4–30.2, body width: 17.8–21.5. Unicolored, in dorsal view, dark brown, pubescent, anterior margin of pronotum reddish brown; in ventral view, head dark reddish brown, legs, abdomen generally reddish brown, margins of abdomen dark brown, tibial spines black. Supra-anal plate transverse type, hind margin nearly straight, median distinctly emarginated, and divided by a longitudinal line. Subgenital plate reddish brown, with median protruded, bulged (Fig. 4 D–G). Nymph. Similar to the female, reddish brown to dark brown, some individuals lighter colored. Ootheca. As Fig. 38 A–B and L. Reddish brown. Serration of keel long, strongly curved, each one’s apex reaching to the other one’s back. Respiratory canals well developed. The arrangement of longitudinal ridges compacted. Variation. Males vary in the following characters: 1) post-clypeus with two large brown spots or not; 2) the whitish yellow margin of pronotum may expand in some individuals, which only with pronotum disc brown, the rest part all whitish yellow; 3) the speckles on the tegmina can be reduced, which with anal and R areas lack brown speckles; some individuals may also have denser maculae, the tegmina strong brownish, which only with small transparent maculae. Natural History. Females and nymphs can be found inside the tree hole in the forest (Fig. 39 A), or under stones (Fig. 39 B–C). Distribution. China (Beijing, Hebei, Henan, Neimenggu, Published as part of Qiu, Lu, Che, Yang-Li & Wang, Zong-Qing, 2018, A taxonomic study of Eupolyphaga Chopard, 1929 (Blattodea: Corydiidae: Corydiinae), pp. 1-68 in Zootaxa 4506 (1) on pages 10-14, DOI: 10.11646/zootaxa.4506.1.1, http://zenodo.org/record/2606718, {"references":["Walker, F. (1868) Catalogue of the specimens of Blattariae in the collection of the British Museum. British Museum, London, 239 pp.","Chopard, L. (1922) Les Polyphaga du groupe du sinensis Walk. [Orth. Blattidae]. Bulletin de la Societe entomologique de France, 194 - 196.","Saussure, H. D. (1869) Melanges Orthopterologiques. Memoires de la Societe de physique et d'histoire naturelle de Geneve, 20, 227 - 328.","Dohrn, H. (1888) Neue und wenig bekannte Blattiden. Stettin Entomological Society, 49, 129 - 132.","Saussure, H. D. (1893) Revision de la tribu des Heterogamiens (Orthopteres de la Famille des Blattides). Revue Suisse de Zoologie, 1, 289 - 318.","Kirby, W. F. (1903) XXXVII. - Notes on Blattidae & c., with Descriptions of new Genera and Species in the Collection of the British Museum, South Kensington. - No. II. The Annals and magazine of natural history, 7, 373 - 381. https: // doi. org / 10.1080 / 00222930308678853","Kirby, W. F. (1904) A Synonymic Catalogue of Orthoptera. Fol. I. Orthoptera Euplexoptera, Cursoria, et Gressoria. (Forficulidae, Hemimeridae, Blattidae, Mantidae, Phasmidae.). British Museum, London, 501 pp.","Chopard, L. (1929) Orthoptera palaearctica critica VII. Les Polyphagiens de la faune palearctique (Orth., Blatt.). Eos, 5, 223 - 358.","Wu, C. - F. (1935) Catalogus Insectorum Sinensium, I. The Fan Memorial Institute of Biology, Peiping, 367 pp.","Bey-Bienko, G. Y. (1950) Fauna of the USSR. Insects. Blattodea, 40. Institute of Zoology, Academy of Sciences of the URSS, Moscow, 343 pp.","Princis, K. (1962) Blattariae: Suborde Polyphagoidea: Fam. Polyphagidae. In: Beier, M. (Ed.), Orthopterorum Catalogus. Pars 3. Uitgeverij Dr. W. Junk, ' s-Gravenhage, 3 - 74.","Woo, F. - C. (1987) Investigations on domiciliary cockroaches from China. Acta Entomologica Sinica, 30 (4), 430 - 438.","Feng, P. - Z., Guo, Y. - Y. & Woo, F. - C. (1997) Cockroaches of China, Species and Control. China Science & Technology Press, 206 pp.","Princis, K. (1959) Revision der Walkerschen und Kirbyschen Blattarientypen in British Museum of Natural History, London. III. Opuscula Entomolagica, 24, 125 - 149."]}

The efficacy of a new and highly virulent Metarhizium anisopliae (Hypocreales: Clavicipitaceae) strain was evaluated against Blattella germanica (L.) (Blattaria: Blattellidae) in the laboratory; this strain was obtained and purified from field-collected Eupolyphaga sinensis cadavers. The status of this fungus as a new and genetically distinct species was supported by ITS sequence comparisons. The new strain was compared with other M. anisopliae isolates and was found to be highly infectious and virulent against B. germanica. The virulence of this new strain against different instars of male and female cockroaches at five conidia concentrations (1 × 105, 1 × 106, 1 × 107, 1 × 108, and 1 × 109 conidia/ml) was evaluated in vitro, and the mortality (measured as lethal concentrations, LC50) was determined. According to the pathogenicity test, M. anisopliae isolate EB0732 produced 100% mortality of one- to three-instar nymph (LC50 = 0.37 × 105conidia/ml) and 78.33% mortality of adult female (LC50 = 1.39 × 107 conidia/ml) at 15 d post-inoculation at a concentration of 1 × 109 conidia/ml. There was an overall significant effect on mortality between the age and sex of B. germanica. A M. anisopliae susceptibility test showed that the survival rate of cockroaches after treatment with topical applications and mixed infection was lower than per os. These studies shed light on a valuable integrated pest management against the German cockroach.

Aim of the study. To examine the antiproliferation and anti-invasion of Eupolyphaga sinensis Walker 70% ethanol extract (ESWE) on breast cancer and elucidate the underlying signaling mechanisms. Methods. MTT and colony formation assays were used to investigate the effect of ESWE on proliferation of breast cancer cells in vitro. The xenograft mouse tumor model was used to determine the effect of ESWE on breast cancer in vivo. To investigate the underlying molecular mechanisms, we used western blotting to analyze the expression of ERK1/2, CXCR4, matrix metalloproteinase 2 (MMP2), and MMP9 pretreated with ESWE. The stromal cell–derived factor (SDF)-1α-induced migration and invasion potential of breast cancer cells were examined by wound-healing assays and Matrigel invasion chamber assays. Results. ESWE effectively inhibited the proliferation of MDA-MB-435s and MDA-MB-231 cells and exhibited antitumor effects in an MDA-MB-231 xenograft mice model. Furthermore, ESWE suppressed the activity of ERK1/2, a key molecule of MAPK signaling. We also observed that ESWE treatment led to downregulation of CXCR4 expression as well as greatly reduced MMP2 and MMP9. ESWE affected CXCR4 expression partially through the modulation of autocrine vascular endothelial growth factor. However, suppression of CXCR4 expression was the result of downregulation of mRNA expression. Inhibition of CXCR4 expression by ESWE further correlated with the suppression of SDF-1α-induced migration and invasion in breast cancer cells. Conclusion. ESWE exerted its antiproliferation and antiinvasion by regulating MAPK signaling and related metastasis factorsand thus could be a useful therapeutic candidate for breast cancer intervention.

Eupolyphaga sinensis Walker has been reported to have anticoagulation, antithrombotic, liver protective and antitumor effects. In the present study, the inhibitory effects on proliferation of A549 human non‑small cell lung cancer cells and the underlying mechanisms were examined. Firstly, three solvents, 70% ethanol, distilled water and 95% ethanol, were used to extract Eupolyphaga sinensis Walker. The MTT assay results demonstrated that the 70% ethanol extract more potently reduced the growth of A549 cells and it was therefore adopted in the subsequent experiments. Eupolyphaga sinensis Walker 70% ethanol extract significantly inhibited A549 cell migration in a time‑ and dose‑dependent manner and inhibited human umbilical vein endothelial cell proliferation, migration and tube formation. Furthermore, Eupolyphaga sinensis Walker 70% ethanol extract effectively inhibited blood vessel formation in the established tissue model for angiogenesis. In addition, Eupolyphaga sinensis Walker 70% ethanol extract was demonstrated to inhibit the autophosphorylation of KDR, and downregulate the subsequent activation of AKT and extracellular signal regulated kinase (ERK)1/2 in A549 cells. In conclusion, these findings demonstrated that the antitumor mechanism of Eupolyphaga sinensis Walker 70% ethanol extract was through inhibiting angiogenesis. It functioned by interrupting the autophosphorylation of KDR and subsequently, AKT and ERK1/2.

An insect trypsin-like enzyme with similar biochemical properties to psychrophilic analogs was purified from Chinese ground beetle (Eupolyphaga sinensis) for the first time. The purified trypsin-like enzyme was designated as E. sinensis trypsin-like enzyme (ESTL) with molecular weight of 22.8 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The maximal activity of ESTL was observed at pH 9.5, and the temperature optimum of ESTL was observed at 45C by its amidolytic effect on the substrate benzoyl-L-Arg-p-nitroanilide. The analysis of inhibitors showed that specific inhibitor of serine proteases (phenylmethanesulfonyl fluoride) and trypsin inhibitors (tosyl–lysine chloromethyl ketone, bovine pancreatic trypsin inhibitor and benzamidine) inactivated ESTL almost completely. Over the range of tested temperature of 10–30C, the catalytic efficiency (kcat/Km) of the ESTL was about five times than that of bovine trypsin. The N-terminal sequencing of ESTL revealed the following sequence: I1VGGSTTTIQ10NFPYQVSL, and the complementary DNA (cDNA) of ESTL was cloned by rapid amplification of cDNA ends. Practical Applications In recent years, increasing attention has been drawn to the psychrophilic (cold-adapted) enzymes. Psychrophilic trypsins with high activity may be interesting for several industrial applications of enzymes, such as in certain food-processing operations that require low-processing temperatures. For example, cod trypsin is already used in food production and cosmetics. In this article, a trypsin-like enzyme [Eupolyphaga sinensis trypsin-like enzyme (ESTL) ] was purified from Chinese ground beetle by a relatively simple method, and the complementary DNA (cDNA) of this enzyme was cloned by rapid amplification of cDNA ends. This enzyme was characterized for molecular and enzymatic properties, as demonstrated that it had high activity at low and moderate temperature compared with the mammalian trypsins, and maintained good thermal stability compared with psychrophilic trypsins. ESTL has potential interest in food industry.

High-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hextane-ethyl acetate-methanol-water (1.5:1:1.5:1, v/v/v/v) was applied to the isolation and purification of attractants from Chinese cockroach, Eupolyphaga sinensis Walker. Two new attractants with attractant activity towards the male insects were obtained from the extract sample in a one-step separation. Their purities were determined by HPLC. Subsequent MS, NMR and CD analyses have led to the characterization of (R)-3-ethyl-6,8-dihydroxy-7-methyl-3,4-dihydroisochromen-1-one (1) and (R)-6,8-dihydroxy-3,7-dimethyl-3,4-dihydroisochromen-1-one (2), two novel isocumarin type attractants. Based on these results, it is concluded that HSCCC is a viable separation method option for purifying insect attractants, while effectively maintaining the attracting activity of the isolates. This is the first attempt to apply counter-current chromatography technique to separate attractants from Chinese cockroach.

Background Termicin is an antimicrobial peptide with six cysteines forming three disulfide bridges that was firstly isolated from the salivary glands and hemocytes of the termite Pseudacanthotermes spiniger. In contrast to many broad-spectrum antimicrobial peptides, termicin is most active against filamentous fungi. Although more than one hundred complementary DNAs (cDNAs) encoding termicin-like peptides have been reported to date, all these termicin-like peptides were obtained from Isoptera insects. Methods The cDNA was cloned by combination of cDNA library construction kit and DNA sequencing. The polypeptide was purified by gel filtration and reversed-phase high performance liquid chromatography (RP-HPLC). Its amino acid sequence was determined by Edman degradation and mass spectrometry. Antimicrobial activity was tested against several bacterial and fungal strains. The minimum inhibitory concentration (MIC) was determined by microdilution tests. Results A novel termicin-like peptide with primary structure ACDFQQCWVTCQRQYSINFISARCNGDSCVCTFRT was purified from extracts of the cockroach Eupolyphaga sinensis (Insecta: Blattodea). The cDNA encoding Es-termicin was cloned by cDNA library screening. This cDNA encoded a 60 amino acid precursor which included a 25 amino acid signal peptide. Amino acid sequence deduced from the cDNA matched well with the result of protein Edman degradation. Susceptibility test indicated that Es-termicin showed strong ability to kill fungi with a MIC of 25 μg/mL against Candida albicans ATCC 90028. It only showed limited potency to affect the growth of Gram-positive bacteria with a MIC of 200 μg/mL against Enterococcus faecalis ATCC 29212. It was inactive against gram-negative bacteria at the highest concentration tested (400 μg/mL). Es-termicin showed high sequence similarity with termicins from many species of termites (Insecta: Isoptera). Conclusions This is the first report of a termicin-like peptide isolated from E. sinensis that belongs to the insect order Blattodea. Our results demonstrate the diversity of termicin-like peptides, as well as antimicrobial peptides in insects.

The insect of Eupolyphaga sinensis Walker has been used as traditional anti-thrombosis medicine without bleeding risk for several hundreds years in eastern countries. Our previous work has identified a bi-functional anti-thrombosis protein containing both direct-acting fibrin(ogen)olytic and plasminogen-activating activities from the insect. By proteomics and transcriptome analysis, 105 serine proteases belonging to four families were identified from the ground beetle, E. sinensis and the classification is for serine proteases of this organism. Pharmacological test indicated that 5 (eupolytin 1-5) of them have the abilities to hydrolyze fibrin(ogen) and/or activate plasminogen. The current work revealed the extreme diversity of anti-thrombosis components in E. sinensis and anti-thrombosis molecular mechanisms of the traditional medicinal insect, and provided many templates for the development of new thrombolytic agents. Especially, these proteins, which contain both plasmin- and PA (plasminogen-activating)-like activities, are excellent candidates for anti-thrombosis medicines.

Body size of insects with flexible life cycles is expected to conform to the saw-tooth model, a model in which the relationship between size and developmental time depends on length of the growing season. In species with high variability in terms of voltinism, however, more complex patterns can be expected. Few empirical studies have demonstrated the existence of such relationships, or whether climatic factors contribute to these relationships. In this study, we investigated the geographic variation in body size of the Chinese cockroach, Eupolyphaga sinensis Walker (Blattaria: Polyphagidae), which has a variable life cycle length. The sizes of adults – collected from 14 localities ranging from temperate to subtropical zones in China – were measured, using body length, body width, and pronotum width as parameters. The relationship between size, latitude, and climate factors (encompassing 10 variables) was then investigated. We found that the body size of E. sinensis varied considerably with latitude: cockroaches were larger at low and high latitudes, but smaller at intermediate latitudes. Thus, the relationship between climate and body size conformed to a saw-tooth pattern. Results indicate that two factors were significantly associated with body size clines: season length and variability in life cycle length. Our results also demonstrated that climatic factors contribute to latitudinal clines in body size, which has important ecological and evolutionary implications. It can be expected that global climate change may alter latitudinal clines in body size of E. sinensis.

Ethnopharmacological relevance Eupolyphaga sinensis Walker popularly known as “preferred drug to regulate blood flow” are traditionally used in folk medicine in the treatment of ecchymoma, posttraumatic wound, hepatic fibrosis and tumor. Aim of the study To characterize chemical compositions and to evaluate the antitumor and immunomodulatory of Eupolyphaga sinensis Walker ethanol extract (ESEE) in hepatocarcinoma H22 bearing mice. Materials and methods ESEE was obtained by ethanol reflux extraction and analyzed by gas chromatography–mass spectrometry (GC–MS) after methylation. ICR mice were treated with ESEE for 14 consecutive days at doses of 31 mg/kg (low-dose), 62 mg/kg (mid-dose) and 124 mg/kg (high-dose) after H22 tumor cells were implanted. At the end of the experiments, the tumor weight of each mouse was measured. Levels of serum TNF-α and IFN-γ was assayed by ELISA. Protein expressions of Bax, Bcl-2 and caspases-3 were detected by immunohistochemistry. Results Chemical analysis revealed the presence of 6 components that account for 97.55% of fatty acids, indicating the occurrence of saturated and polyunsaturated fatty acids. Oral administration of ESEE could inhibit tumor growth, promote Th1 type cytokine productions (TNF-α and IFN-γ) and induce apoptosis of hepatocarcinoma via increase of Bax/Bcl-2 ratio and activation of caspases-3. Oral administration of ESEE in a dosage of 6.2 g/kg did not lead to toxic effects in mice. Conclusions ESEE was effective in inhibiting tumor growth in vivo and could also serve as immunoadjuvant for tumor therapy.

The responses of organisms to temperature variations may be via short term responses of the phenotype (phenotypic plasticity), or they could involve long-term evolutionary change and adaptation (via selection) to the genotype. These could involve changes to the mean size of the animal or to the thermal reaction norm. We examined the effects of various temperatures (of 22, 25, 28, and 31 °C) on development time, adult body size and preadult survivorship in three populations of the cockroach, Eupolyphaga sinensis (Walker), collected at different latitudes. We found substantial temperature-induced plasticity in development time, body size, and preadult survivorship, indicating that developmental temperatures have strong impacts on growth and life history traits of E. sinensis. Genetic differences for development time, body size, and preadult survivorship were detected among populations, and the three traits exhibited highly significant variations in the responses of different populations to various temperature conditions, indicating genetic differences among populations in terms of thermal reaction norms. We also found that two populations seem to support the beneficial acclimation hypothesis whereas the third mid-latitude population does not. The results are likely because of differences in season length and voltinism, indicating that not only temperature regime but also its interactions with generation time (and development time), voltinism, and season length are likely to have considerable effects on insect development time and body size. Overall, changes in development time, body size, and preadult survivorship in E. sinensis can all be regarded as adaptations to changing thermal regimes.

To screen stimulators from Chinese medicinal insects for mycelial growth and polysaccharides production of Ganoderma lucidum, G. lucidum was inoculated into the media with and without supplementation of medicinal insect extracts. The ethyl acetate extract of Eupolyphaga sinensis at 55 mg l(-1) lead to significant increase in both biomass and intracellular polysaccharides (IPS) concentration from 8.53 +/- 0.41 to 14.16 +/- 0.43 and 1.28 +/- 0.09 to 2.13 +/- 0.11 g l(-1), respectively. In addition, the ethyl acetate extract of Catharsius molossus at 55 mg l(-1) significantly enhanced extracellular polysaccharides (EPS) production; the EPS yield increased from 350.9 +/- 14.1 to 475.1 +/- 15.3 mg l(-1). There were no new components in the two types of polysaccharides obtained by the addition of the insect extracts.

Ethnopharmacological relevance Yaotongning (YTN) is a traditional Chinese Medicine (TCM) that contains ten component medicinal materials (CMMs) and uses Chinese rice wine as a vehicle to enhance its efficacy. YTN has been used for rheumatoid arthritis (RA) treatment in China for decades and has been reported to have anti-inflammatory and analgesic effects, as well as to strengthen the immune system. Aim of the study The present work quantitatively evaluated the in vitro effects of active fractions from the ten CMMs that make up YTN and eight additional herbs commonly used in TCM formulas for RA treatment, as well as different combinations of these active fractions, on cellular immune response; the findings were used to determine which active fractions are responsible for promoting an immune response, and to assess whether YTN is superior to other similar formulas and whether YTN can be improved by simplifying its formula from the point of its cellular immunomodulatory activity. Materials and methods Using the YTN formulation principles and some concepts in combinatorial chemistry, 27 TCM samples were designed by combining some or all of the active fractions of YTN and other eight herbs used for RA treatment. Release of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) from ANA-1 murine macrophages was measured using an enzyme-linked immunosorbent assay (ELISA). The immunoregulatory effects of the TCM samples were evaluated by comparing their half-effective concentrations (EC 50 ) for stimulating the release of these cytokines. Results Among the investigated active fractions, the flavonoids from Carthamus tinctorius (Fct), Davallia mariesii (Fdm), and Cinnamomum cassia Twig volatile oils (Vca) from the eight selected herbs effectively promoted IL-1β and IL-6 release from ANA-1 cells. Saponins from the YTN CMM Glycyrrhiza uralensis (Sgu) were the most potent promoters of IL-1β and TNF-α release. The aqueous extract of YTN CMM Eupolyphaga sinensis (Ves) strongly enhanced the release of IL-1β, IL-6, and TNF-α from ANA-1 cells. The EC 50 values for stimulating the secretion of IL-1β, IL-6, and TNF-α could be determined for only six samples. The full YTN formula and the sample containing 50% Glycyrrhiza uralensis saponins, 25% of the mixture of alkaloids, and 25% of the mixture of all flavonoids exhibited good comprehensive cellular immunomodulatory activity. The immunomodulatory activity of the complete YTN formula was better than that of the sample containing all active fractions of YTN without Chinese rice wine (the YTN vehicle). Conclusions Sgu and Ves are the primary active fractions of YTN involved in stimulating immune responses. The YTN prescription was reasonably effective at promoting cellular immune responses. Chinese rice wine, the YTN vehicle, strengthened the immunoregulatory activity of YTN. The results of this study demonstrate that the YTN recipe could be improved by reducing the number of CMMs and altering some active fractions without reducing its activity to promote cellular immune responses.

The complete mitochondrial genome of Polyphaga plancyi is a 15,547 bp circular molecule, which contains 37 typical mitochondrial genes (13 protein-coding genes, 2 rRNAs and 22 tRNAs) and a 854 bp D-loop. Its gene arrangement pattern is identical with typical other cockroaches. All protein-coding genes start with an ATG codon except COI, ND3, ND5, ND4, ND4L ND6 and ND1. TAA is the most frequent stop codon, and TAG, GCA, TA- and T- are also occurred very common. The mtDNA sequence contains 12S rRNA and 16S rRNA of rRNA. Except for tRNASer(AGY) and tRNALeu(CUN) without the dihydrouridine (DHU) arm, all tRNAs could be folded into canonical cloverleaf secondary structures. The phylogenetic trees from the ML and BI analyses based on the complete mtDNA of nine cockroaches’ species, share similar topologies and high node support values. Polyphaga plancy has close relative with Eupolyphaga sinensis.

Tumor growth and metastasis are responsible for most cancer patients' deaths. Here, we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis. Compared with proliferation, colony formation, transwell assay and transplantable tumor in nude mouse in vitro and vivo, eupolyphaga sinensis walker extract (ESWE) showed good inhibition on the SMMC-7721 cell growth and metastasis. Using genome-wide microarray analysis, we found the down-regulated growth and metastasis factors, and selected down-regulated genes were confirmed by real-time PCR. Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE. Moreover, our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ, Akt, m-TOR, Erk1/2, MEK-2, Raf and JNK-1, but also increasing cyclin D1 protein levels and decreasing amount of cyclin E, cyclin B1 and cdc2 of the cycle proteins. At the same time, ESWE reduced MMP2, MMP9 and CXCR4, PLG, NFκB and P53 activities. Overall, our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC, AKT, MAPK signaling and related metastasis signaling, having potential in cancer therapy.

Eupolyphaga sinensis Walker is not only used as a food to enhance immunity, but is used as a traditional Chinese medicine and is known as the "preferred drug to regulate blood flow". Previous studies have reported its potential biological activities including anticoagulation, antithrombotic, liver protective effect and antitumor effects. Our results indicated that E. sinensis Walker 70% ethanol extract exhibited anti-tumor effects on S180 (murine sarcoma cell line) cells implanted mice. It effectively inhibited K562 (human chronic myeloid leukemia cell line) cells proliferation and induced G(2)-M phase arrest accompanying through up-regulation of cyclin B1, cdc2 and down-regulation of cyclin D1, cyclin E1, cdc25c and p53. In addition, it inhibited EGF secretion and EGFR kinase activity. Western blotting analysis indicated that it also inhibited the phosphorylation EGFR and activation of its downstream signaling molecules AKT and ERK. These results suggested that the antitumor mechanism of E. sinensis Walker involved altering the cell cycle and inhibiting EGFR phosphorylation in the EGFR signaling pathway.

Background MicroRNAs are now recognized as key post-transcriptional regulators in animal ontogenesis and phenotypic diversity. Eupolyphaga sinensis Walker (Blattaria) is a sexually dimorphic insect, which is also an important source of material used in traditional Chinese medicine. The male E. sinensis have shorter lifecycles and go through fewer instars than the female. Furthermore, the males have forewings, while the females are totally wingless. Results We used the Illumina/Solexa deep sequencing technology to sequence small RNA libraries prepared from the fourth-instar larvae of male and female E. sinensis. 19,097,799 raw reads were yielded in total: 7,817,445 reads from the female library and 11,280,354 from the male, respectively. As a result, we identified 168 known miRNAs belonging to 55 families as well as 204 novel miRNAs. Moreover, 45 miRNAs showed significantly different expression between the female and the male fourth-instar larvae, and we validated 10 of them by Stem-loop qRT-PCR. Some of these differentially expressed miRNAs are related to metamorphosis, development and phenotypic diversity. Conclusions/Significance This is the first comprehensive description of miRNAs in E. sinensis. The results provide a useful resource for further in-depth study on molecular regulation and evolution of miRNAs. These findings not only enrich miRNAs for hemimetabolans but also lay the foundation for the study of post-transcriptional regulation on the phenomena of sexual dimorphism.

OBJECTIVE To obtain the cDNA sequence encoding fibrinolytic enzyme from Eupolyphaga sinensis and express it in prokaryotic and eukaryotic expression system. METHOD The primers were designed according to the cDNA of other animals'fibrinolytic enzyme. The cDNA sequence was cloned by RT-PCR and 3 RACE. RESULT Sequence analysis revealed that the length of the cDNA fragment was 672 bp and encoded a protein of 224 amino acid residues, the N end amino acid sequence residues was IVGG in accordance with other fibrinolytic enzyme. The cDNA sequence was expressed in E. coli, inactive protein was obtained. While expressed in Pichia pastoris, recombinant protein had fibrinolytic activity. CONCLUSION The cDNA sequence of fibrinolytic enzyme from E. sinensis Walker was cloned and expressed for the first time and it proved a good basis for further functional study of the enzyme.

We present the complete mitochondrial DNA sequence of Eupolyphaga sinensis. This closed circular molecule is 15553 bp long and consists of 37 genes that encode for 13 inner membrane proteins, 2 ribosomal RNAs and 22 transfer RNAs. The genome shares the gene order and orientation with previously known Blattaria mitochondrial genomes. All tRNAs could be folded into the typical cloverleaf secondary structure, but the tRNASer (AGN) appears to be missing the DHU arm. The A + T-rich region is 857 bp long and longer than other cockroaches. Based on the concatenated amino acid sequences of all protein coding genes of E. sinensis in conjunction with those 23 other arthropod sequences, we reconstruct the phylogenetic tree. Phylogenetic analyses shows that Blataria (including Isoptera) and the Mantodea are sister groups. Furthermore the relationship of the three basal clades of winged insects are different from the three previous hypotheses ((Ephemeroptera + Odonata) +Neoptera, Ephemeroptera + (Odonata + Neoptera), Odonata + (Ephemeroptera +Neoptera)). The Ephemeroptera (Parafronurus youi) clusters with the Plecoptera (Pteronarcys princes).

To screen stimulators for mycelial growth and polysaccharides production of Ganoderma lucidum , the fungus was inoculated into the media with and without supplementation of several fatty acids and ether extracts of Chinese medicinal insects. Palmitic acid at 1.0 g/L lead to significant increase in both biomass and extracellular polysaccharides (EPS) concentration from 7.73 plusmn 0.41 to 10.21 plusmn 0.43 g/L and 342.6 plusmn 10.7 to 401.3 plusmn 14.9 mg/L, respectively. In addition, ether extract of Eupolyphaga sinensis at 1.5 g/L also was of great advantage to biomass and EPS production. There were no new components in the EPS obtained by the addition of palmitic acid.

Generally the "Zhe-chong" (〓虫) is a Chinese crude drug of Eupolyphaga sinensis WALKER or two other kinds of insects which belong to Blattidae. But in Formosa Dytiscidae insects are used as "Zhe-chong." Once, in a certain place in China, "Zhe-chong" included both Dytistidae and Blattidae insects, but today what is implied is limited to the latter. In literature, various views are expressed on what is the original insects of crude drug "Long-shi" (竜蝨). In order to explain these doubtful points, we identified materials purchased in Hong-Kong market and proved the origin.