This study compared the chemical profiles, component content, dry paste yield, and pharmacological effects of samples obtained from the mixed single decoctions and the combined decoction of Gegen Qinlian Decoction(GQD), aiming to provide an experimental foundation for evaluating the equivalence of the two decocting methods and the suitability of TCM formula granules in clinical application. The same decoction process was used to prepare the combined decoction and mixed single decoctions of GQD. Ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was employed to compare the chemical profiles between the two groups. High-performance liquid chromatography(HPLC) was used to compare the content of nine characteristic components between the two groups. Then, a delayed diarrhea mouse model induced by irinotecan was established to compare the pharmacological effects of the two groups on chemotherapy-induced diarrhea. The UPLC-Q-Exactive Orbitrap MS in ESI~+ and ESI~- modes identified 59 chemical components in the compound decoction and mixed single decoctions, which showed no obvious differences in component species. The content of baicalin and wogonoside was higher in the compound decoction, while that of puerarin, daidzein-8-C-apiosylglucoside, berberine, epiberberine, wogonin, glycyrrhizic acid, and daidzein was higher in the mixed single decoctions. Further statistical analysis revealed no significant difference in the content of the nine characteristic components between the compound decoction and the mixed single decoctions. The dry paste yield had no significant difference between the two groups. Compared with the model group, both compound decoction and mixed single decoctions alleviated the weight loss and reduced diarrhea index in mice. Both of them lowered the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), cyclooxygenase-2(COX-2), intercellular adhesion molecule-1(ICAM-1), interleukin-10(IL-10), malondialdehyde(MDA), and nitric oxide(NO) in the colon tissue. Furthermore, they significantly increased the levels of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD). Hematoxylin-eosin(HE) staining showed that colon tissue cells were tightly arranged with clear nuclei in both groups without obvious difference. The compound decoction and mixed single decoctions showed no significant differences in chemical component species, content of nine characteristic components, dry paste yield, or the pharmacological effects on alleviating chemotherapy-induced diarrhea. The findings provide a reference for evaluating the flexibility and superiority of combined or single decocting method in the preparation of TCM decoctions or formula granules.
Animals ; Mice ; Biological Products ; Chromatography, High Pressure Liquid ; Coleoptera ; Cyclooxygenase 2 ; Diarrhea/drug therapy* ; Antineoplastic Agents

Poria(Fu Ling) is a bulk traditional Chinese medicine(TCM)with a long history and complex varieties. The royal medical records of the Qing Dynasty include multiple medicinal materials of Fu Ling, such as Bai Fu Ling(white Poria), Chi Fu Ling(rubra Poria), and Zhu Fu Ling(Poria processed with cinnabaris). The Palace Museum preserves 6 kinds of specimens including Fu Ling Ge(dried Poria), Bai Fu Ling, Chi Fu Ling, Zhu Fu Ling, Bai Fu Shen(white Poria cum Radix Pini), and Fu Shen Mu(Poria cum Radix Pini). After trait identification and textual research, we found that Fu Ling Ge was an intact sclerotium, which was processed into Fu Ling Pi(Poriae Cutis), Bai Fu Ling and other medicinal materials in the Palace. The Fu Ling in the Qing Dynasty Pa-lace was mainly from the tribute paid of the officials in Yunnan-Guizhou region. The tribute situation was stable in the whole Qing Dynasty, and changed in the late Qing Dynasty. The cultural relics of Fu Ling in the Qing Dynasty Palace confirm with the archival documents such as the royal medical records and herbal medicine books, providing precious historical materials for understanding Fu Ling in the Qing Dynasty and a basis for the restoration of the processing of the Fu Ling in the Qing Dynasty Palace.
Animals ; Poria ; China ; Books ; Coleoptera ; Medical Records ; Wolfiporia

The present study collected data on traditional Chinese medicine(TCM) compounds effective in relieving pain from the patent database of the State Intellectual Property Office(SIPO), sorted out the TCM compounds against pain in patents, and analyzed the medication rules to provide references for the research and development of new TCM drugs against pain. The data were subjected to frequency statistics, association rules, cluster analysis, and complex network analysis by IBM SPSS Modeler 18.3 and SPSS Statistical 26.0. The results showed that among the 101 oral prescriptions included in the statistics, the top 5 drugs were Glycyrrhizae Radix et Rhizoma, Angelicae Sinensis Radix, Paeoniae Radix Alba, Chuanxiong Rhizoma, and Salviae Miltiorrhizae Radix et Rhizoma, and among the 49 external prescriptions included in the statistics, the top 5 drugs were Myrrha, Olibanum, Angelicae Dahuricae Radix, Borneolum Syntheticum, and Chuanxiong Rhizoma. Whether oral or external prescriptions, the drugs were mainly warm in nature, and bitter, pungent, and sweet in flavor. According to TCM complex network analysis, the core drugs were Glycyrrhizae Radix et Rhizoma, Angelicae Sinensis Radix, Paeoniae Radix Alba, and Chuanxiong Rhizoma in oral prescriptions, and Olibanum, Myrrha, Glycyrrhizae Radix et Rhizoma, Chuanxiong Rhizoma, and Angelicae Sinensis Radix in external prescriptions. Overall, the therapeutic principles of oral prescriptions were mainly replenishing Qi, nourishing blood, and promoting Qi and blood circulation, while those of external prescriptions were activating blood, resolving stasis, promoting Qi flow, and relieving pain on the basis of the oral prescriptions. In the future research and development of TCM compounds against pain, the prescriptions should be modified with mind-tranquilizing and depression-relieving drugs. With the modernization of TCM, the development of new pain-relieving TCM compound patents based on ancient methods and clinical experience adhering to the guidance of TCM treatment based on syndrome differentiation can meet the new demand for pain treatment in the current society and give full play to the advantages of TCM in pain treatment.
Animals ; Medicine, Chinese Traditional ; Frankincense ; Pain ; Paeonia ; Coleoptera

Objective To establish the basic data for estimating minimum postmortem interval （PMImin） of heavily decayed and skeletonized remains by studying the development of Dermestes maculatus DeGeer （Coleoptera: Dermestidae）. Methods The developmental stages of Dermestes maculatus were observed at four constant temperatures of 20 ℃, 24 ℃, 28 ℃ and 32 ℃, and the changes in body length were also examined as the biological indicator to estimate larval day-age and instar. Results The total developmental time from egg to adult at 20 ℃, 24 ℃, 28 ℃ and 32 ℃ were （126.7±10.6） d, （69.4±8.2） d, （50.4±8.4） d and （49.6±6.5） d, respectively. The body length increased gradually, but changed irregularly as a whole. Conclusion The study provides basic data on the development and growth of Dermestes maculatus, especially on its developmental duration as a significant value for estimating PMImin of heavily decayed and skeletonized remains. Nevertheless, the change of body length is not found to be the best biological indicator for instar determination.
Animals ; Autopsy ; Coleoptera ; Larva ; Temperature

Aims: Oryctes rhinoceros beetle is one of the most damaging pests of oil palm and cause high oil palm mortality. The empty fruit bunch mulch and rotten old trunk of oil palm in the field provide the organic matter for the breeding sites and increases the number of O. rhinoceros larvae. Bacillus thuringiensis as bioinsecticide can synthesize crystal proteins toxic to the larvae. The present study was aimed to find effective B. thuringiensis isolates as biopesticide against O. rhinoceros larvae. Methodology and results: Screening process was carried out through heating of soil sample suspension at 80 °C to eliminate the non-spore formers and plated onto T3 medium. Colony morphology was observed, followed by Gram and endospore staining. The crystal protein was observed by Coomassie Brilliant Blue (CBB) staining. Bioassay test was conducted by force-feed method followed by food contamination method. The results showed isolates SBB33 and SBB35 were able to infect and caused high mortality to the O. rhinoceros larvae. Isolates SBB33 and SBB35 showed the highest mortality against 1st instar larvae (94.44% and 75% respectively) and 3rd instar larvae (64.8% and 60% respectively) compared to control treatments. The 16S rRNA gene sequencing showed SBB33 has high similarity with B. thuringiensis strain 3S2-3, while SBB35 has high similarity with B. thuringiensis strain GCU_BTi10. Protein separation of the spore-crystal mixture by SDS-PAGE showed the prominence of 66 kDa protein band that was predicted to be Cry toxins which is specific to coleopterans insect. Conclusion, significance and impact of study Bacillus thuringiensis isolates SBB33 and SBB35 have high potential as biopesticides against O. rhinoceros larvae and could be used to control major pests in oil palm plantation.
Bacillus thuringiensis--isolation & ; purification ; Coleoptera

Insect antifreeze protein (AFP) has high antifreeze activity. Antifreeze proteins can be used in cryopreservation of biological tissues and cells. We expressed an antifreeze protein from the desert beetle Microdera punctipennis in yeast and determined the function of the protein at low temperatures. Yeast expression vector, pPIC9K-Mpafp698, was constructed and transformed into Pichia pastoris GS115. The expression of MpAFP698 was induced by methanol, and identified by tricine SDS-PAGE and Western blotting. Mpafp698 gene was inserted into the genome of the host yeast strain GS115, and correctly expressed. Hardly any yeast's own protein was secreted into the media. Cryoprotective experiments showed that MpAFP698 can significantly protect mouse liver as well as other mouse organs from cold damage compared with those in the control of Bovine serum albumin (BSA) addition. Besides, the hemolysis of blood cells protected by MpAFP698 at 4 degrees C was reduced and the survival rate of SF9 cells protected by MpAFP698 after freezing and thawing was increased compared to those of the control with BSA addition. Our results showed that MpAFP698 can be expressed in yeast, which allows a convenient purification of the MpAFP protein that has the cryoprotective effect.
Animals ; Antifreeze Proteins ; biosynthesis ; Blotting, Western ; Cold Temperature ; Coleoptera ; Cryoprotective Agents ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Freezing ; Insect Proteins ; biosynthesis ; Mice ; Pichia ; metabolism ; Sf9 Cells

With the deterioration of environment, and the excessive collection of wild resources, the wild populations of Myla- bris phalerata Pallas are less and less, almost extincted in many traditional distribution areas. It is necessary to breed M. phalerata artificially for sustainable utilization. Food preference of adult M. phalerata is the key to its provenance screening and domestication in the artificial breeding. In this paper, the food preference of 3 geographical populations of M. phalerata was studied. The results showed that the food preferences of adult M. phalerata in different geographical populations were different. The adult M. phalerata in Wuming preferred cucumber flowers, gourd flowers and melon flowers. The adult M. phalerata in Tianlin preferred cowpea flowers. And the adult M. phalerata in Guangzhou preferred cowpea flowers and gourd flowers. Gourd flowers were the most attractive food for the adult M. phalerata of 3 geographical populations of M. phalerata.
Animals ; Averrhoa ; Breeding ; Coleoptera ; physiology ; Cucurbitaceae ; Flowers ; Food ; Food Preferences ; physiology ; Geography

OBJECTIVE: To explore the growing development and community succession of main sarcosaphagous insects on pig carcasses in summer indoor and outdoor environment in Shenzhen area and to estimate the postmortem interval (PMI). METHODS: From early May to August in 2013, in Forensic Medical Examination Center of Shenzhen Public Security Bureau, the main insect species and the decomposition process were observed in two adult pig carcasses of simulative indoor and outdoor environment. The different decomposition stages and the community succession of insects were recorded. RESULTS: The indoor and outdoor pig carcasses showed skeleton 412.5 and 325 hours after death, respectively. The main species of flies on pig carcasses were Chrysomya megacephala, Chrysomya rufifacies and Chrysomya chani. The main species of beetles were Crecphilus maxillosus, Necrobia ruficollis, Saprinus splendens and Dermestes maculatu. The dominant species of flies in the outdoor pig carcasses obviously produced the second generations due to the effect of mass rainfall, nor in the indoor pig carcasses. CONCLUSION There are regular patterns on the community succession of insects on pig carcasses in summer indoor and outdoor environment in Shenzhen area. The activity patterns of seven typical insects and their larva show important value for estimating PMI.
Animals ; Autopsy ; Cadaver ; China ; Coleoptera ; Death ; Diptera ; Environment ; Insecta/growth & development* ; Larva ; Population Dynamics ; Postmortem Changes ; Seasons ; Swine

OBJECTIVEThe aim of the present study was to screen the Metarhizium strains with high virulence against the larvae of Dorysthenes hydropicus, a serious pest of Citrus grandis.

METHODThirty six strains of Metarhiziums were isolated from the soil of C. grandis GAP base and collected from other institutions, and the pathogenicity of these strains against 1st instar larvae of D. hydropicus was detected at concentration of 1 x 10(8) conidia/g. The high violence strains against D. hydropicus were cultivated in sabouraud dextrose yeast medium at first, then transfer to rice grain. And the sporulations of these violent strains against D. hydropicus were detected.

RESULTTwenty-eight strains showed virulence against D. hydropicus by preliminary study, and 7 strains of them were collected for further study, 6 of the 7 showed high virulence, the highest cadaver rate was higher than 74%. The conidia production of strain 1 and strain 4 were 2.35 +/- 0. 25 (1 x 10(9) conidia/g), 2.21 +/- 0.27 (1 x 10(9) conidia/g), respectively, showed significantly higher than other strains.

CONCLUSIONStrain 1 and strain 4 of the 36 Metarhiziums strains showed high virulence against D. hydropicus, and the highest sporulation ability, so they have a best application prospect.

Animals ; Citrus ; parasitology ; Coleoptera ; microbiology ; Metarhizium ; growth & development ; isolation & purification ; pathogenicity ; Pest Control, Biological ; Plant Diseases ; parasitology ; prevention & control ; Soil Microbiology ; Spores, Fungal ; growth & development ; isolation & purification ; pathogenicity ; Virulence

OBJECTIVE: To document the arthropod succession pattern and to identify forensically important species in northeastern Egypt (32° 15' E and 30° 36' N) for the first time. METHODS: Carcasses were exposed in an open area for 60 days during summer season. Ambient daily temperature (maximum and minimum) and relative humidity (RH) were recorded and existing keys were used for identification of different species. RESULTS: During the period of study, the mean of maximum and minimum temperatures were 34.85 °C and 29.2 °C respectively, while the mean of RH was 53.5%. Four stages of decomposition were observed: fresh, bloat, decay and dry. The most abundant orders were found to be Diptera, Coleoptera and Hymenoptera. Arthropods were collected belonging to 4 families of Diptera: Muscidae, Fanniidae, Calliphoridae and Sarcophagidae. While there were 2 families of Coleoptera: Dermestidae and Histeridae. Monomorium species was the only Hymenoptera family in this study. CONCLUSION The present work provided a basis for further studies dealing with insect colonization of carcasses in different seasons and locations in Egypt.
Animals ; Arthropods ; classification ; physiology ; Coleoptera ; Diptera ; Egypt ; Entomology ; Feeding Behavior ; Forensic Medicine ; methods ; Hymenoptera ; Insecta ; classification ; Rabbits ; Rats ; Seasons ; Temperature