Research Advances in Pharmacological Action and Clinical Application of Periplaneta americana

Researches on pharmacological action and drug development of Periplane- ta americana have demonstrated that it has wide medicinal value, low side effects and rich resources, having good application prospects in medicine field. This article reviewed the researches on pharmacological action and clinical application of Periplaneta americana in order to provide references for the further research and devel- opment of Periplaneta americana medicines.

一亚热带蜚蠊(Periplaneta japanna)两地理种群间相异的光周期反应模式(英文)

为弄清光周期对一种亚热带蜚蠊(Periplaneta japanna)若虫发育的控制机理,检测了光周期对冲绳(26°N)和八丈岛(33°N)两个地理种群若虫发育的影响。结果表明,光周期显著影响两地理种群的若虫发育,然而,冲绳和八丈岛种群间显示不同的光周期反应模式。冲绳种群在长日条件下(LD16∶8h)若虫发育显著快于中间日长(LD14∶10h)和短日条件下(LD12∶12h)的若虫发育,孵化后60d或120d将若虫自LD16∶8h向LD12∶12h转移,若虫发育受到抑制,而自LD12∶12h向LD16∶8h转移,则促进若虫发育。八丈岛种群的若虫在短日条件下发育最快,其次是中间日长,长日条件下若虫发育最慢,孵化后60d或120d将若虫自LD16∶8h向LD12∶12h转移抑制若虫发育,反方向转移的若虫快速发育。两地理种群间不同的光周期反应模式可以认为是趋异进化的结果。

美洲大蠊(Periplaneta americana L.)抗菌肽杀菌作用初步观察

报道了美洲大蠊(PeriplanetaamericanaL.)抗菌肽对大肠杆菌(EscherichiacoliK88)的作用过程.电镜观察表明,美洲大蠊抗菌肽首先使细菌的外层及细胞质膜损伤,形成开口,导致内容物外泄而死亡,最后菌体崩解成碎片.

Periplaneta americana extract used in patients with systemic inflammatory response syndrome

BACKGROUND:Periplaneta americana extract is recognized to have a positive effect on gastrointestinal mucosa.This study aimed to investigate the effects of periplaneta americana extract on immune function,nutrition status and gastrointestinal complications of early enteral nutrition patients with systemic inflammatory response syndrome(SIRS).METHODS:Patients with SIRS were randomly divided into two groups:treatment and control groups.All patients in the two groups received conventional therapy including enteral nutrition,but periplaneta americana extract,an additional Chinese medicine,was given to the patients in the treatment group.At the beginning of treatment(0 day)and 1,3,and 7 days after treatment,the levels of immunoglobulin(Ig A),total lymphocyte count(TLC),total protein(TP)and prealbumin(PA)were respectively tested in patients'venous blood.The incidences of bloating,diarrhea,aspiration pneumonia and high blood sugar at 7 days after treatment were recorded.The mortality of the patients in 28 days was recorded.RESULTS:At 3 and 7 days after treatment,the levels of Ig A and TLC in the treatment group were higher than those in the control group(P<0.05).At 7 days after treatment,the levels of TP and PA in the treatment group were higher than those in the control group(P<0.05).The incidences of bloating and diarrhea in the treatment group were lower than those in the control group,the differences were significant(P<0.05).The mortality of treatment group was lower than that of the control group(P>0.05).CONCLUSION:Periplaneta americana extract could reduce gastrointestinal complications and improve immune function and nutritional status in patients with systemic inflammatory response syndrome.

Periplaneta Americana Extract Ameliorates LPS-induced Acute Lung Injury Via Reducing Inflammation and Oxidative Stress

Objective Acute lung injury(ALI)is an acute clinical syndrome characterized by uncontrolled inflammation response,which causes high mortality and poor prognosis.The present study determined the protective effect and underlying mechanism of Periplaneta americana extract(PAE)against lipopolysaccharide(LPS)-induced ALI.Methods The viability of MH-S cells was measured by MTT.ALI was induced in BALB/c mice by intranasal administration of LPS(5 mg/kg),and the pathological changes,oxidative stress,myeloperoxidase activity,lactate dehydrogenase activity,inflammatory cytokine expression,edema formation,and signal pathway activation in lung tissues and bronchoalveolar lavage fluid(BALF)were examined by H&E staining,MDA,SOD and CAT assays,MPO assay,ELISA,wet/dry analysis,immunofluorescence staining and Western blotting,respectively.Results The results revealed that PAE obviously inhibited the release of proinflammatory TNF-α,IL-6 and IL-1βby suppressing the activation of MAPK/Akt/NF-κB signaling pathways in LPS-treated MH-S cells.Furthermore,PAE suppressed the neutrophil infiltration,permeability increase,pathological changes,cellular damage and death,pro-inflammatory cytokines expression,and oxidative stress upregulation,which was associated with its blockage of the MAPK/Akt/NF-κB pathway in lung tissues of ALI mice.Conclusion PAE may serve as a potential agent for ALI treatment due to its anti-inflammatory and anti-oxidative properties,which correlate to the blockage of the MAPK/NF-κB and AKT signaling pathways.

Comparison between Decolorization Effects of Activated Carbon and Macroporous Resin on Periplaneta americana L. Skimmed Cream

[Objective] The aim of this study was to investigate the decolorization ef- fects of activated carbon and macroporous resin on Periplaneta americana L. skimmed cream and compare the advantages and disadvantages of the two decolorization technologies. [Method] Periplaneta americana L. skimmed cream was decolored with activated carbon and macroporous resin, and freeze-dried to collect solid decolorization products. By investigating the yield, decolorization rate, protein retention rate and decolorization operation process, the advantages and disadvantages of the two decolorization technologies were compared. [Result] Both activated carbon and macroporous resin can be used for decolorization. To be specific, macroporous resin-decolorization is superior in the yield and protein retention rate, while activated carbon-decolorization is superior in decolorization rate and decolorization operation process. [Conclusion] Macroporous resin-decolorization can be used if protein is the main ingredient required in the experiment, while activated carbon-decolorization can be used if protein is not the main ingredient required.

Exploration on Separation of Periplaneta Americana Small Peptides with Tricine-SDS-PAGE System

[ Objective] This study aimed to explore a Tricine-SDS-PAGE method for separation and purification of Periplaneta Americana small peptides. [ Method] By comparing the separation effects of three types of gels with different ratios on P. Americana small peptides, an appropriate Tricine-SDS-PAGE method for separation of small peptides was established. [ Result] Standard low molecular weight Marker was preliminarily separated to a certain extent with polyacrylamide gel electraphoresis, while the sample mixture was not separated due to the different levels of aggregation in separating gel, spacer gel and stacking gel. [Condusion] More appropriate polyacrylamide gel electrephoresis separation method or other better separation methods were to be further explored for separating peptide mixture with smaller molecular weight.

Separation of Antitumor Active Plypeptides from Periplaneta Americana by Ultra Filtration Technology

[Objective] The paper was to establish ultrafiltration membrane technology for separation of antitumor active plypeptides from Periplaneta Americana. [Method] Hollow fiber ultrafiltration membrane（3 kDa）was used to separate antitumor active plypeptides from P. Americana. Through single factor test, the impact of rotation speed, concentration ratio, pH and operation time on separation effort was studied, and the separation technology conditions were further optimized through orthogonal test. [Result] The best technology conditions for separation of antitumor active plypeptides from P. Americana were as follows： rotation speed 4 500 r/min, concentration ratio 30%, pH 11, and the rotation speed was the major influencing factor. [Conclusion] The study laid foundation for industrialization of ultrafiltration membrane separation of antitumor active plypeptides from P. Americana.

Potential Risk of Transmission of Escherichia coli and Salmonella spp. Infections by the Musca domestica Fly and the Periplaneta americana Cockroach in the City of Cotonou (South Benin)

Background: The occurrence of bacterial infections sometimes involves synanthropic flies and cockroaches, as mechanical vectors of pathogenic microorganisms. The aim of this study was to identify the different species of flies and cockroaches that cohabit with humans in the city of Cotonou and determine the contribution of Musca domestica and Periplaneta americana in the transmission of bacteria responsible for infections in humans. Methods: Capture sessions during the day for flies and at night for cockroaches were carried out in 4 arrondissements of the city of Cotonou. The insects collected were transported to the Center of Research Entomological of Cotonou for identification, then sent to the Section Hygiene of Water and Food for microbiological analysis, enumeration and testing for human pathogenic bacteria. Results: A total of 351 flies and 26 cockroaches were collected at the sites, including two synanthropic species belonging to two families each. Musca domestica and Periplaneta americana were the most abundant species found after identification in the city of Cotonou. Thermo-tolerant coliforms and Escherichia coli were found on all specimens at varying concentrations. Salmonella spp. bacteria were identified in the fly population from the Dantokpa dump. Conclusion: Flies and cockroaches can transmit microorganisms to humans. The presence of Salmonella spp. among the specimens revealed that infections frequently caused by contaminated food or water are also transmitted to humans by flies. The presence of these germs on flies and cockroaches represents a potential risk of mechanical transmission to humans. It is, therefore, essential to continue investigations in order to assess the species of bacteria propagated, improve control strategies against these troublesome insects and adopt better hygiene conditions for better living.

黑胸大蠊(Periplaneta fuliginosa)病毒的分离及某些特性

从黑胸大蠊(Periplaneta fuliginosa)自然罹病的虫尸中分离得到一株非包涵体病毒。将病毒悬液均匀拌入无菌饲料并供食154～169日龄黑胸大蠊健康若虫时,能使其感染、发病,死亡率可达98％以上。在电子显微镜下观察时,病毒为球形二十面体颗粒,直径约23nm。病毒悬液具有典型核蛋白紫外吸收光谱。病毒用DNase和RNase处理并经吖啶橙染色、二苯胺和苔黑酚试验及甲醛反应证明:该病毒含有单链DNA。以上特性与细小病毒科的特征有点类似。

Periplaneta americana extract promotes infectious diabetic ulcers wound healing by downregulation of LINC01133/SLAMF9

Wound healing in diabetic ulcers remains a significant clinical challenge,primarily due to bacterial infection and impaired angiogenesis.Periplaneta americana extract(PAE)has been widely used to treat diabetic wounds,yet its underlying mechanisms are not fully understood.This study aimed to elucidate these mechanisms by analyzing long non-coding RNA(lncRNA)expressions in the wound tissues from diabetic anal fistula patients treated with or without PAE,using high-throughput sequencing.Peripheral blood monocytes from patients were differentiated into M0 macrophages with human macrophage colony-stimulating factor(hMCSF)and subsequently polarized into M1 macrophages with lipopolysaccharide.The results indicated that LINC01133 and SLAMF9 were downregulated in wound tissues of patients treated with PAE.Furthermore,PAE suppressed M1 macrophage polarization and enhanced human umbilical vein endothelial cell(HUVEC)proliferation,migration,and angiogenesis.These effects were diminished when LINC01133 or SLAMF9 were overexpressed.Mechanistically,LINC01133 was shown to upregulate SLAMF9 through interaction with ELAVL1.Overexpression of SLAMF9 reversed the effects of LINC01133 silencing on macrophage polarization and HUVEC functions.In conclusion,PAE facilitates the healing of infected diabetic ulcers by downregulating the LINC01133/SLAMF9 pathway.

Protective effect of periplaneta americana extract on intestinal mucosal barrier function in patients with sepsis

OBJECTIVE： To investigate the effect of the periplaneta americana extract on the intestinal mucosal barrier and prognostic implications in patients with sepsis. METHODS： Sixty and six patients with sepsis were assigned randomly to treatment group （32 cases） and control group （32 cases）. The extractfrom periplaneta americana plus conventional medication for sepsis was administered to the treatment group, while the control group only received conventional treatment. The gastrointestinal function scores and acute physiology and chronic health evaluation II （APACHE 11 ） scores of all subjects were documented at baseline, at days 1, 3 and 7 after treatment respectively and their blood endotoxin was tested at the same time points as well. The incidence of death was recorded for both groups throughout the trial. RESULTS： At days 3 and 7 after treatment, gastrointestinal function score, APACHE II, and endotoxin level in treatment group wasbetter than that in con- trol group and the difference between them was significant （both P〈0~05）. Although the incidence of death in treatment group was less than that in control group, the difference between the two groups was not significant （P〉 0.05）. CONCLUSION： The extract of periplaneta ameri- cana had protective effect on intestinal mucosal barrier and could improve the condition and prog- nosis in patients with sepsis.

Molecular and functional characterization of the American cockroach, Periplaneta americana, Rab5： the first exopterygotan low molecular weight ovarian GTPase during oogenesis

The small Rab GTPases are key regulators of membrane vesicle trafficking. Ovaries of Periplaneta americana （Linnaeus） （Blattodea： Blattidae） have small molecular weight GTP/ATP-binding proteins during early and late vitellogenic periods of oogenesis. However, the identification and characterization of the detected proteins have not been yet reported. Herein, we cloned a cDNA encoding Rab5 from the American cockroach, P. americana, ovaries （PamRab5）. It comprises 796 bp, encoding a protein of 213 amino acid residues with a predicted molecular weight of 23.5 kDa. PamRab5 exists as a single-copy gene in the P. americana genome, as revealed by Southern blot analysis. An approximate 2.6 kb ovarian mRNA was transcribed especially at high levels in the previtellogenic ovaries, detected by Northern blot analysis. The muscle and head tissues also showed high levels of PamRab5 transcript. PamRab5 protein was localized, via immunofluorescence labeling, to germline-derived cells of the oocytes, very early during oocyte differentiation. Immunoblotting detected a ~25 kDa signal as a membrane-associated form revealed after application of detergent in the extraction buffer, and 23 kDa as a cytosolic form consistent with the predicted molecular weight from amino acid sequence in different tissues including ovary, muscles and head. The PamRab5 during late vitellogenic periods is required to regulate the endocytotic machinery during oogenesis in this cockroach. This is the first report on Rab5 from a hemimetabolan, and presents an inaugural step in probing the molecular premises of insect oocyte endocytotic trafficking important for oogenesis and embryonic development.

Systemic RNAi of V-ATPase subunit B causes molting defect and developmental abnormalities in Periplaneta fuliginosa

The vacuolar (H+)-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B in the smokybrown cockroach Periplaneta fuliginosa (Blattodea). We observed a high homology score of the deduced amino acid sequences between 10 species in seven orders. RNAi of the vha55 gene in R fuliginosa caused nymphal/nymphal molting defects with incomplete shedding of old cuticles, growth inhibition, as well as bent and wrinkled cuticles of thoraxes and abdominal segments. Since growth inhibition caused by vha55 RNAi did not interfere in the commencement of cockroach molting, molting timing and body growth might be controlled by independent mechanism. Our study suggested V-ATPases might be a good candidate molecule for evolutionary and developmental studies of insect molting.

New insight into foregut functions of xenobiotic detoxification in the cockroach Periplaneta americana

The physiological functions of insect foregut,especially in xenobiotic detox ification,are scarcely reported because of unimportance in appearance and insufficient molecular information.The cockroach Periplaneta americana,an entomological model organism,provides perfect material to study physiological functions of foregut tissue due to its architectuxal feature.Through Illumina sequencing of foregut tissue from P.amer icana individuals (control)or insects treated with cycloxaprid,as a novel neonicotinoid insecticide,54 193 166 clean reads were obtained and further assembled into 53 853 unigenes with an average length of 366 bp.Furthermore,the number of unigenes involved in xenobiofic detoxification was analyzed,mainly including 70 cytochrome P450s,12 glutathione S-transferases (GSTs),seven carboxylesterases (CarEs)and seven adenosine triphosphate-binding cassette (ABC)transporters.Compared to control,the expression of 22 xenobiotic detoxification unigenes was up-regulated after cycloxaprid application, mainly containing 18 P450s,one GST,two CarEs and one ABC adenosine triphosphate transporter,indicating that the oxidation-reduction was the major reactive process to cy cloxaprid application.Through quantitative real-time polymerase chitin reaction analysis, the expression of selected unigenes (six P450s,one GST and one CarE.)was up-regulated at least two-fold following cycloxaprid treatment,and was generally in agreement with transcriptome data.Compared to the previous midgut transcriptome of P.americana,it looks like the expressive abundance of the xenobiotic detoxification unigenes might be important factors to the detoxifying fimctional differences between foregut and midgut.In conclusion,insect foregut would also play important roles in the physiological processes related to xenobiotic detoxification.

美洲大蠊研粉对脊髓损伤大鼠氧化应激的影响

目的探讨美洲大蠊研粉对脊髓半横断损伤(hSCI)大鼠脊髓损伤区域氧化应激的影响。方法60只雌性SD大鼠按照随机数字表法分为对照组和观察组,各30只,两组均通过打开T10椎板行左侧脊髓半横断,制作hSCI模型。术前30 min及术后1~14 d,对照组予0.9%氯化钠溶液、观察组予美洲大蠊研粉(630 mg/kg)溶于0.9%氯化钠溶液灌胃,5 mL/只,每天1次。术后6 h及1、3、7、14 d每组每个时间点取5只大鼠,用酶联免疫吸附试验检测丙二醛(MDA)、8-羟基脱氧鸟苷(8-OHdG)含量;术后14 d每组取5只大鼠用于脊髓组织尼氏染色。结果与对照组比较,观察组在显微镜10×20倍及10×40倍视野下脊髓组织神经元数目均更多[(26.25±0.50)个比(10.84±0.58)个、(14.92±0.32)个比(6.75±0.32)个,P<0.01];观察组术后7、14 d脊髓组织MDA[(0.61±0.03)nmol/mL比(0.89±0.02)nmol/mL、(0.54±0.01)nmol/mL比(0.68±0.02)nmol/mL,P<0.01]和8-OHdG[(17.92±0.99)ng/mL比(23.37±0.77)ng/mL、(15.91±0.28)ng/mL比(20.18±0.37)ng/mL,P<0.01]含量均低于对照组;与对照组比较,观察组氧化应激高峰时间更早,MDA和8-OHdG表达的峰值提前到了术后3 d。结论美洲大蠊研粉降低了hSCI大鼠脊髓损伤区域MDA、8-OHdG的表达,使氧化应激高峰时间提前,减轻了氧化应激损伤,减少了神经元丢失。

Complete nucleotide sequence and genomic organization of Periplaneta fuliginosa densonucleosis virus

We have cloned the replicative form of the Periplaneta fuliginosa densonucleosis virus (PfDNV) genome and determined its complete sequence. The sequence has 5 454 nucleotides (nt), the genome consists of an internal unique sequence flanked by inverted terminal repeats (201 nt). The first 122 nt at the 5’end and the terminal 122 nt at the 3’end of both plus and minus strands can fold into a typical hairpin structure. The genome contains seven major open reading frames (ORFs). The plus strand has 4 ORFs occupying the 5’half of the plus strand, whereas the others span the 5’ half of the minus strand. Two potential promoters were found at map units (m.u.) 3 and 97. Computer analysis of sequence homologies with other parvoviruses suggests that the plus strand of Pf DNV encodes very likely the nonstructural proteins and the minus strand probably encodes the structural proteins.

Comparative analysis of the three-dimensional structure of Periplaneta fuliginosa densovirus

The three-dimensional structure of Periplaneta fuliginosa densovirus (pfDNV) is determined at 2.3 nm reso-lution using the techniques of cryo-electron microscopy and image reconstruction. The pfDNV contains five structural proteins and 60 protein subunits arranged on a T = 1 icosa-hedral shell with a relatively smooth surface. Its recon-struction reveals its distinct capsid structure from those ob-served in CPV and GmDNV. As in GmDNV, spike-like protrusions are not present in pfDNV at the threefold axes; while two small thorn-like protrusions are identified there. However, different from CPV and GmDNV, cylindrical channels along the fivefold axes are closed in pfDNV; while a small thorn-like protrusions, which have not been reported in other parvovirus, are observed there in pfDNV although their function is yet to be investigated. The pfDNV has dim-ple-like depressions at the icosahedral twofold axes; but has no canyon-like regions encircling the fivefold axes. The ico-sahedrally well-ordered nucleic acid has also been observed in pfDNV, suggesting that the protein and nucleic acid probably form closed interaction.

一测多评法测定美洲大蠊药材5种核苷类成分

目的建立高效液相色谱(HPLC)一测多评法同时测定美洲大蠊中尿嘧啶、尿苷、次黄嘌呤、肌苷及鸟苷5种成分的含量。方法采用Agilent ZORBAX SB-Aq色谱柱(250 mm×4.6 mm,5μm);以甲醇-0.01 mol·L^(-1)磷酸二氢钾溶液为流动相,梯度洗脱;流速:1.0 mL·min^(-1);柱温:20℃;检测波长:260 nm;进样量:10μL。计算内参物尿苷与其他成分的相对校正因子(fa/b)。一测多评法测定10批美洲大蠊中5种成分含量,并与外标法比较。结果5种核苷类在各自范围内线性良好(r>0.9995),平均加样回收率97.0%~100.8%。尿嘧啶、次黄嘌呤、肌苷、鸟苷的fa/b分别是0.9080、1.0053、1.9695、1.3034。结论所建立的方法准确稳定,可为美洲大蠊药材及提取物的质量控制提供理论参考。

美洲大蠊肠道菌株WA5-1-7的初步鉴定及其抗菌活性成分的研究

目的对具有抗白念珠菌活性的美洲大蠊肠道来源菌株WA5-1-7进行形态学和分子生物学鉴定,并对其次级代谢物进行分离纯化,从中筛选具有抗菌活性的单体化合物。方法采用染色法和扫描电子显微镜观察菌株形态;利用PCR技术扩增菌株16S rRNA,通过其基因序列与NCBI数据库进行BLAST比对并采用N-J法构建系统发育树。菌株发酵后经乙酸乙酯溶剂萃取获得粗提物,粗提物经硅胶柱层析、葡聚糖凝胶柱层析、HPLC分离纯化,获得单体化合物,经核磁共振、质谱数据与文献比对鉴定化合物结构,并检测化合物的抗菌活性。结果菌株WA5-1-7初步鉴定为环圈链霉菌(Streptomyces anulatus),从粗提物中分离出10个化合物,分别为SF2738F(1)、SF2738D(2)、环(脯氨酸-亮氨酸)二肽(3)、SF2738A(collismycin A,4)、环(苯丙氨酸-脯氨酸)二肽(5)、SF2738C(6)、1H-吲哚甲醛(7)、大豆黄酮(8)、放线菌素X2(9)、放线菌素D(10),其中化合物4、9和10有较强抗菌活性。结论本研究从美洲大蠊肠道来源链霉菌次级代谢产物中分离出具有抗菌活性的单体化合物,为深入探索美洲大蠊肠道菌奠定了基础。