Phytochemical Screening and Acute oral Toxicity Study of Root Extracts of Combretum hereroense Schinz and Balanites aegyptiaca Del. Traditionally Used to Treat Female Infertility in Baringo County, Kenya

Aims: To carry out phytochemical screening and acute oral toxicity test to validate their safety and efficacy. Study Design: Standard phytochemical screening tests were used to highlight phytochemical compounds of roots of the plants. The evaluation of acute toxicity of the root extracts of the plants followed the model of Acute Toxicity Class based on OECD 423 Guideline, 2001. Place and Duration of the Study: The study was undertaken at the Department of Chemistry & Biochemistry for the extraction for samples extraction and phytochemical screening. Acute oral toxicity studies were done at the Department of Biological Sciences for acute toxicity study, University of Eldoret, Between June and September 2022. Methodology: Phytochemical screening for presence of Tannins, saponins, flavonoids, glycosides, alkaloids, anthocyanin, terpenoids, steroids, coumarins, lipids, proteins and carbohydrates were carried out. Acute oral toxicity studies were done using the fixed dose method at a dose of 2000mg/kg body weights of rats. Three groups were used: control and test groups for each of the respective plant root extracts. Signs of toxicity and/or mortality were monitored daily for 14 days. Weekly fasting body weights were also recorded. Results: The phytochemical screening results showed the presence of tannins, saponins, flavonoids, glycosides, alkaloids, anthocyanin, terpenoids, steroids, lipids, proteins and carbohydrates present in the root extract of Combretum hereroense. Tannins, saponins, flavonoids, glycosides, terpenoids, steroids, and carbohydrates were present in root extracts of Balanites aegyptiaca. Following the acute oral toxicity study, there were no abnormalities observed in physiological parameters. In addition, no deaths were recorded during the study period. The LD50 was therefore greater than 2000 mg/kg. The fasting body weights of extract treated rats increased stably compared to the control [p = .05]. Conclusion: The results showed C. hereroense and B. aegyptiaca methanol root extracts were considered safe in acute oral exposure. Long-term toxicity studies are needed for further toxicological profile elicitation of the plant, and a possible reinforcement of clinical relevance of the results of laboratory studies.

Application progress of air-liquid interface exposure technology in respiratory system toxicity induced by diesel exhaust in vitro / 环境与职业医学

Diesel exhaust (DE) is an important pollution source widely existing in the living and production environment, which is closely related to the health of the public and occupational groups. The International Agency for Research on Cancer has classified DE as a Group 1 carcinogen. Considering the negative health impacts on the respiratory system due to DE exposure in vitro, it is crucial to apply reliable test systems allowing accurate assessment of the biological effects of DE. The exposure technology of respiratory system in vitro is considered as one of the feasible measures to implement the 3R (reduce, refine, and replace) principle in animal experiments. Compared with the traditional submerged culture in vitro models, the air-liquid interface (ALI) exposure technology has the advantages including fewer influencing factors, easier exposure condition control, and shorter exposure cycle. ALI has become an important tool to study molecular events associated with physiology and pathology of respiratory system, and action modes and interactions of different cell types. Also, ALI has been increasingly widely used because it can simulate the actual processes of human respiratory system cells and/or tissues to DE exposure. This review was intended to introduce the development and advantages of ALI exposure technology, and further summarized the application progress of ALI exposure technology in studying the respiratory toxicity induced by DE exposure in vitro, so as to provide new ideas and pathways for the use of ALI exposure technology in the study of biomarkers and mechanisms of respiratory toxicity associated with DE exposure, and provide basic data to screen and promote biomarkers for exposed populations.

Bacillus thuringiensis A21, un aislado no tóxico de elevada actividad larvicida residual contra Aedes aegypti / Bacillus thuringiensis A21, a non-toxic isolate of high residual larvicidal activity against Aedes aegypti

Introducción: Los productos derivados de Bacillus thuringiensis se han empleado con éxito en el control de insectos. Sin embargo, la poca residualidad promueve la búsqueda de alternativas. En Cuba diferentes estudios informan la evaluación y selección de aislamientos de B. thuringiensis con actividad larvicida contra Aedes aegypti: el aislado A21 se destaca por su elevada actividad larvicida y ausencia de betaexotoxina. Objetivo: Evaluar la toxicidad/patogenicidad aguda oral y actividad larvicida residual del aislado A21. Métodos: La evaluación de la toxicidad/patogenicidad aguda oral del aislado A21 se estableció en el modelo biológico Rattus norvegicus. Se realizaron observaciones clínicas diarias de los animales y se evaluó el peso corporal. Se estimó la eliminación y la infectividad de B. thuringiensis mediante análisis de muestras de heces, y de fluidos y órganos, respectivamente. Para determinar la actividad larvicida residual del aislado A21 frente a larvas de Ae. aegypti se realizaron bioensayos con diferentes volúmenes de recambios de agua semanal (total, parcial, no recambio) en los recipientes. La mortalidad obtenida se calculó a las 24-72 h. Resultados: Con el aislado A21 no se detectó toxicidad/patogenicidad aguda oral en Rattus norvegicus. En el estudio de residualidad, la mortalidad larvaria a las 72 h se mantuvo elevada (80-100 por ciento) hasta la semana 24. A partir de la semana 25 la mortalidad larvaria disminuyó (p < 0,05). Conclusiones: Se evidencia la baja toxicidad y la elevada actividad larvicida residual del aislado A21 contra Ae. aegypti y lo convierten en un candidato promisorio para el desarrollo de biolarvicidas. Estos productos biológicos podrían contribuir a mejorar las estrategias de control del vector existentes en Cuba(AU)

ABSTRACT Introduction: Bacillus thuringiensis-based products have been successfully used for insect control. However, their low residuality promotes the search for alternatives. In Cuba, different studies have informed about the evaluation and selection of B. thuringiensis isolates with larvicidal activity against Aedes aegypti: isolate A21 highlights for its high larvicidal activity and absence of beta-exotoxins. Objective: To evaluate the acute oral toxicity/pathogenicity and the residual larvicidal activity of isolate A21. Methods: The evaluation of the acute oral toxicity/pathogenicity of isolate A21 was established in the animal model Rattus norvegicus. Daily clinical observations of the animals were carried out, and their body weight was evaluated. The elimination and infectivity of B. thuringiensis were estimated by analyzing feces, and fluids and organs samples, respectively. To determine the residual larvicidal activity of isolate A21 to Ae. aegypti larvae, bioassays were conducted with different volumes of weekly water changes (total, partial, no change) in the containers. Mortality was calculated at 24h-72h. Results: No acute oral toxicity/pathogenicity was detected with isolate A21 in Rattus norvegicus. In the study of residuality, at 72h larval mortality remained high (80 percent-100%) until week 24, decreasing from week 25 (p < 0.05). Conclusions: It is evidenced the low toxicity and high residual larvicidal activity of isolate A21 against Ae. aegypti, which makes it a promising candidate for the development of biolarvicides. These biological products could contribute to the improvements of the existing vector control strategies in Cuba(AU)

Biological properties of Schinus terebinthifolia Raddi essential oil

Abstract Schinus terebinthifolia Raddi green fruits essential oil (EO) was evaluated regarding its phytochemical profile, antimicrobial and cytotoxic activities, and toxicity. Gas chromatography with mass spectrometry was applied to identify its constituents, thereafter the minimum inhibitory concentration, minimum bactericidal and fungicidal concentrations, and its antibiofilm activity were evaluated. The EO cytotoxicity was assessed in tumor and non-tumor human cells, and in vivo toxicity was evaluated in a Galleria mellonella model. The major constituents of S. terebinthifolia EO were alpha-phellandrene and beta-phellandrene. The EO had a weak activity against all strains of Candida albicans (MIC 1000µg/mL) and had no activity against non-albicans strains, bacteria, and C. albicans biofilm. Cytostatic activity against all tumor cell lines was shown. Additionally, cell viability remained at EO concentrations up to 62.5 µg/mL. At 16 mg/mL, 50% hemolysis was observed, and it had low toxicity in vivo. Overall, the S. terebinthifolia EO was characterized by low antimicrobial and antibiofilm activities, with no evidence of toxicity to human tumor and non-tumor cells

Synthesis of a new type of carbon dots by pyrolysis of human hair using motor oil as a dispersant and their biological effects / 中草药

Objective: New type of nano-carbon dots were found after pyrolysis of human hair using motor oil as a dispersant and the biological effect of these carbon dots was evaluated by animal experiments. Methods: High-temperature pyrolysis was used to carbonize human hair and motor oil, and the carbonized products were extracted, filtered, and dialyzed to obtain a new type of water-soluble substance, carbon dots, named JYRF-CDs. JYRF-CDs were characterized using transmission electron microscopy (TEM) and high-resolution TEM, as well as ultraviolet-visible, fourier transform infrared, fluorescence spectroscopy and X-ray photoelectron spectroscopy (XPS). CCK-8 toxicity test using RAW264.7 cells was used to evaluate the safety of JYRF-CDs and the biological effects of the JYRF-CDs were evaluated by mouse ear swelling experiments and mouse acetate writhing experiments. Results: These JYRF-CDs were nearly spherical and well separated from each other, with a size distribution range of 1.8-3.6 nm, the CDs had a lattice spacing of 0.219 7 nm. The results of cytotoxicity experiments showed that JYRF-CDs had low toxicity, and the results of animal experiments showed that JYRF-CDs had good anti-inflammatory and analgesic effects. Conclusion: In this study, new type of carbon dots, JYRF-CDs, were discovered after pyrolyzing human hair with motor oil as a dispersant for the first time. Taking JYRF-CDs as a breakthrough, the material base of carbonization products after pyrolysis of human hair by high-temperature pyrolysis using motor oil as a dispersant was more clearly explained, providing a new method for the research of nano compounds.

Teratogenic effects of aqueous extract of Ficus glomerata leaf during embryonic development in zebrafish (Danio rerio)

Over the past few decades, many reports were published in scientific journals describing medicinal properties of Ficusglomerata (FG). However, its effects on embryonic development and its safety characteristics have not been studied.The purpose of this investigation was to determine lethal concentration 50 (LC50) and study the effect of aqueousextract of FG leaf (AEFG) on developmental abnormalities in zebrafish embryos. LC50 value of AEFG was calculatedby using probit analysis. Effect on percentage hatchability, heartbeat rate, total body length, and developmentalmorphological abnormalities, i.e., delayed growth, abnormal movement, tail detachment, abnormal head-trunkangle, scoliosis/flexure, and yolk sac edema were recorded. AEFG revealed LC50 of 239.88 ppm. The result showeda significant reduction in percent hatchability (p < 0.05), heartbeat rate (p < 0.001), total body length (p < 0.001),and developmental morphological abnormalities in the embryos treated with AEFG. This research can be used inconsidering the safety of an AEFG extract for their use during pre-conception or early pregnancy period.

Comparison of the Quality of Generic and Original Preparations of Thioctic Acid Injection and Study on Its Acute Toxicity Test in Mice / 中国药房

OBJECTIVE： To compare the quality of original preparations of Thioctic acid injection and generic preparations from 2 domestic manufacturers， discuss the acute toxicity test of mice and to investigate the project of consistency evaluation methods. METHODS： According to the quality standard that stated in Chinese Pharmacopeia， physicochemical properties （characters， pH， osmotic pressure， etc.， contents and related substances of samples of Thioctic acid injection as well as LD50 in acute toxicity test （n＝10）， and mortality of mice after administration of injection solution （n＝30） were compared among 3 manufacturers. RESULTS： The physicochemical properties as and related substances of the original drug and 2 generic drugs were all in line with the quality standard； the contents of 3 samples ranged 95％-105％. The acute toxicity test results showed that the LD50 values of 2 generic drugs （LD50： 247.911 mg/kg， 95％ confidence interval： 222.209-277.999 mg/kg；LD50： 215.291 mg/kg， 95％ confidence interval： 196.637-235.053 mg/kg） were smaller than that of original drug （LD50： 266.534 mg/kg， 95％ confidence interval： 250.597-283.418 mg/kg）， but there was no statistical difference （P＞0.05）. The results of 3 repeated experiments showed that there was statistical significance in the number of animal death caused by the 2 generic drugs （26， 28） was more than that of the original drug （19） （all P＜0.05）， when injection solution was injected into mice in a single dose. After administration of the original drug， mice showed excitatory reactions such as movement and squeal， while 2 generic drugs showed inhibitory reactions. CONCLUSIONS： 2 generic drugs of Thioctic acid injection and the original drug all conform to the relevant regulations of Chinese Pharmacopoeia in terms of preparation quality standards， but the results of acute toxicity test are quite different， so it is difficult to prove the consistency between the 2 generic drugs and the original drug. Therefore， acute toxicity test is necessary for the consistency evaluation of injections.

Efeitos tóxicos de compostos de vanádio sobre os parâmetros biológicos de embriões e adultos de zebrafish (Danio rerio) / Toxic effects of vanadium compounds on biological parameters of embryos and adults of zebrafish (Danio rerio)

Foram avaliados os efeitos tóxicos do metavanadato de sódio (MV), pentóxido de vanádio (PV) e sulfato de oxovanádio (SV), potenciais fármacos antidiabéticos, em embriões e adultos de zebrafish (Danio rerio). Os embriões foram expostos a concentrações de 10-1000µg/mL para avaliação da CL50 96h e seus efeitos teratogênicos. Os adultos foram expostos a 10 e 20µg/mL dos mesmos compostos para se avaliarem alterações comportamentais relacionadas à exposição química e à mortalidade. A CL50 96h foi de 22,48, 53,62 e 74,14µg/mL para MV, SV e PV, respectivamente. Houve 100% de mortalidade nas concentrações de 400-1000µg/mL dos três compostos. Os efeitos teratogênicos mais observados (P<0,05) nos embriões foram edemas de pericárdio e saco vitelínico. Foram constatados, nos animais adultos expostos aos compostos de vanádio, maior batimento opercular e congestão nos arcos branquiais. A exibição dos comportamentos Flutuar e Descansar nos adultos expostos foi significativa (P<0,05), como também a exibição do comportamento Respiração Aérea. Pode-se concluir que a exposição química aos compostos de vanádio causou efeitos tóxicos em embriões e adultos de zebrafish com alta mortalidade. Diante disso, o seu uso como potencial fármaco antidiabético deve ser mais bem estudado em razão do efeito tóxico dessas substâncias.(AU)

The toxic effects of sodium metavanadate (MV), vanadium pentoxide (PV) and oxovanadium sulfate (SV), potential antidiabetic drug, on embryos and adults of zebrafish (Danio rerio) were evaluated. Embryos were exposed to concentrations of 10-1000µg/mL for evaluation of 96-h LC50 and their teratogenic effects. Adults were exposed to 10 and 20µg/mL of the same compounds to evaluate behavioral changes related to chemical exposure and mortality. The 96-h LC50 were 22.48, 53.62, and 74.14µg/mL for MV, SV, and PV, respectively. Mortality of 100% was observed at the concentrations of 400-1000µg/mL of the three compounds. The teratogenic effects most observed (P<0.05) were pericardial and yolk sac edemas. Adult animals exposed to the vanadium compounds had higher opercular beats and congestion in the gill arches. The exhibition of behaviors Floating and Resting in the exposed adults was significant (P<0.05), as well as the Air breathing behavior. Chemical exposure to vanadium compounds caused toxic effects in embryos and adults of zebrafish with high mortality. In conclusion, its use as a potential antidiabetic drug should be better studied due to the toxic effect.(AU)

Expression, purification and characterization of diphtheria toxin mutant CRM197 in Eschrichia coli / 生物工程学报

CRM197 (cross-reacting material 197), a non-toxic mutant of diphtheria toxin, has wide application potential in biopharmaceuticals. However, it is difficult to express CRM197 in bacteria other than Corynebacterium diphtheriae. Here we proposed a new alternative method to produce soluble CRM197 without label in Escherichia coli. In particular, a synthetic gene coding for CRM197, optimized for E. coli codon usage, was cloned in the pET32a (+) vector. Accordingly, the over-expression of the protein was simply induced with IPTG in E. coli BL21 (DE3). The target protein was soluble and accounted for about 40% of the total protein in the supernatant. Following an ultrasonic cytolysis step, the recombinant protein was purified by anion exchange, affinity and desalting chromatography and the purity of the final preparation reached 95%. Cytotoxicity tests showed that the IC₅₀ value of CRM197 was 2.1×10⁷ times the IC₅₀ value of diphtheria toxin, and 9.6 times the IC50 value of diphtheria toxoid, telling that the target protein is safe and non-toxic. Subsequently, we found that both the high dose (20 μg) and the low dose (2 μg) of CRM197 were equally efficient in inducing an immune response against diphtheria toxiod in mice, and the antibodies titer of mice after three immunizations with low dose could reach 1:409 600. In conclusion, our findings provide a highly efficient strategy for the rapid production and purification of unlabeled and soluble recombinant CRM197 in E. coli, with good immunogenicity and safety.

Toxicity study of three roots granule medicine / Монголын эм зүй, эм судлал

Purpose@#To study the effect of Three roots granule medicine with compound medicines ingredient of Asparagus cohinchinensis, Polygonatum odoratum and Polygonatum sibiricum on acute toxicity test and genotoxicity tests. @*Methods@#In toxicity study of Three roots granule medicine, by acute toxicity test were observed general status of animals, body weight changes, signs of poisoning and death for 14 day and determined the maximum tolerated dose, by Ames test, mouse bone marrow polychromatic erythrocyte micronucleus test and mouse sperm deformity test were determined genotoxicity effect. The data were analyzed through SPSS 19.0.@*Results and Conclusions@#In the result of toxicity study, three roots granule medicine was MTD>15g/kg, no acute toxic activity, did not induce mutagenic effect in Ames test and was negative in mouse bone marrow polychromatic erythrocyte micronucleus test and mouse sperm deformity test. Three roots granule medicine has no acute toxicity effect, no genotoxicity effect and safety. We as regard as in future can continuously study to the other pharmacology study of three roots granule medicine.

A toxicological evaluation on the safety of the Eucommia folium / 预防医学

Objective To evaluate the safety of the Eucommia folium and to provide the basis of the rational use ofEucommia folium resource.Methods According to < Technical Standards for Testing & Assessment of Health Food>,version 2003, acute safety test/micronucleus test of born marrow in mice/sperm shape abnormality test in mice/Ames test andthirty-day feeding test were all performed after water extract concentrations of Eucommia folium treatment (1 mLconcentrations equal to 1.5 g Eucommia folium) . Results On the acute toxicity test, the MTD of rats and mice were both40.0 mL/kgbw. The results of genetic toxicity test were all negative in the dosage of 10 g/kg bw(according to the dosage ofEucommia folium) .The results of thirty-day feeding test indicated that there have no significant differences in body weight,blood cytology indexes, blood biochemical indexes, organ/body ratio and pathematology examination in the rats in all thegroups (0.83, 1.67, 3.30 mL/kg) as compared to the control group(P>0.05) . Conclusion Under this experimentalcondition, no obvious abnormity and toxic reaction were observed.

Cardiotoxicity and neurotoxicity features of Aconiti Radix ethanol extract and its impacts on vascular endothelial / 中草药

Objective: To investigate the features of cardiotoxicity and neurotoxicity and the influence on vascular endothelial of ethanol extract of Aconiti Radix (AR). Methods: Totally 45 Wistar rats were randomly divided into four groups, and ig given low-, middle-, and high-dose 70% ethanol extract of AR (in the raw drug dose of 2.5, 5.0, and 7.5 g/kg respectively) for 15 d. The toxic reaction and death condition were observed during administration. The electrocardiogram (ECG) change was tested after administration. Blood was collected for blood routine and blood biochemistry analysis. Brain was harvested for calculating viscera index and pathological examination. Left ventricular and hippocampus were extracted and stained with electron microscope (EM) methods. Results: At the end of treatment, all rats were sacrificed in the high-dose AR group. The mortality and hematologic parameters including RBC, HGB, HCT, ALT, UREA, and GLU were significantly increased in the middle-dose AR groups compared with control group. Arrhythmia especially ventricular arrhythmia occurred in low- and middle-dose AR group. Brain index and number of pyknotic neurons in focal area of hippocampus were significantly increased in middle-dose AR group. EM examination indicated that dissolution of neuronal inclusions, nuclear fragmentation, and coagulation necrosis of glial cells were prominent in the hippocampus tissue in middle-dose AR group. In the middle-dose AR group, the vascular endothelial cells injury and apoptosis were obviously observed in left ventricular and hippocampus separately. Conclusion: The ethanol extract of AR can lead to cardiotoxicity and neurotoxicity especially vascular endothelial injury in dose-dependent manner.

Safety Evaluation of Methoxyflavones Mixture (Ⅱ) from Kempferia parviflora：28-day Repeated Dose Oral Toxicity Test / 日本補完代替医療学会誌

Kaempferia parviflora rhizome is used folk medicines for treatment of various symptoms in Thailand since anticent times.Several types of methoxyflavones has been identified in this plant and their physiological functions have been reported.We determined that six kinds of methoxyflavones (5,7,3',4'-tetramethoxyflavone, 3,5,7,3',4'-pentamethoxyflavone, 5,7-dimethoxyflavone, 5,7,4'-trimethoxyflavone, 3,5,7-trimethoxyflavone, 3,5,7,4'-tetramethoxyflavone) were included in the 80% ethanol extract of K. parviflora rhizome.The safety of six methoxyflavones mixture was evaluated with 28-day repeated oral dose toxicity test in mice.These results indicated no significant toxicity on body weight, blood analyses, organ weight, blood biochemical analyses.

Two-Week Repeated Dose Toxicity of Atractylodis Rhizoma Alba in F344 Rats

This research is to estimate the toxicity of Atractylodis Rhizoma Alba (ARA) in F344 rats and to find a dose level for the 13 weeks toxicity study. A hot water extract of ARA (ARWE) was administered orally to F344 rats at dose levels of 0 (vehicle control), 500, 1000, 2000, 3500, and 5000 mg/kg/day for 2 weeks. Each group was composed to five male and five female F344 rats. According to the result, there were no ARWE-related adverse changes in mortality, body weights, food consumption, urinalysis, hematology, clinical chemistry, gross finding at necropsy, and organ weight examination. Salivation was observed in 3500 and 5000 mg/kg/day in male and female rats but it could not have found any relationship with ARWE administration. Based on our findings, ARWE may not cause toxicity in rats under the experimental conditions. Therefore, dose level of 5000 mg/kg/day as a highest treatment group in 13-week exposure study is recommended for further toxicity assessment.

Preparation and acute toxicity test of Bruceae Fructus oil and β-cyclodextrin polymer inclusion complexes / 中草药

Objective: To prepare Bruceae Fructus oil (BFO) β-cyclodextrin polymer (CDP) inclusion complexes (BFO-CDP-IC), and investigate its safety by acute toxicity test. Methods: BFO was extracted and CDP was prepared according to the literature, the IC of BFO with CDP (BFO-CDP) was prepared by homogenizing method and characterized by SEM, 1H-NMR, and FT-IR. The optimum preparation process was determined by single factor test and L9(34) orthogonal test. Entrapment efficiency (EE) was determined by HPLC. Acute toxicity of BFO-CDP-IC was assessed by determining the number of deaths of ICR mice over gavage treatment for 2 weeks. The commercial emulsion of BFO (BFOE) was used as a reference. Results: The extraction rate of BFO was 17.3%, the yield of CDP was 49.26%, and the degree of crosslinking was 8.47. The optimal conditions for preparation were as follows, ratio of BFO and CDP was 1:10, preparation temperature and time was 20℃ and 6 min, the amount of ether was 0.5 g. Additionally, the HPLC data showed that drug loading of complexes was 7.1%, and EE was 80%. In the acute toxicity test, the median lethal dose (LD50) of BFOE was 9.78 g/kg. In contrast, all mice treated with IC survived even at the highest dosage (15.36 g/kg). Conclusion: The prepared BFO-IC has high EE compared with commercially available BFOE, BFO-CDP polymer significantly decreased toxicity of BFO.

Safety Study of Polyisobutylene Gutong Plaster by Transdermal Administration / 中国药房

OBJECTIVE:To investigate the safety of Polyisobutylene (PIB) Gutong plaster by transdermal administration. METHODS:66 rabbits were randomly divided into a normal group,a group with intact skin and a group with damaged skin. The latter two groups were respectively re-divided into PIB group,the groups of low,medium and high-dose PIB Gutong plaster and Gutong plaster group. An acute toxicity test was conducted on the rabbits,which 14 d of continuous observation was made 24 h af-ter transdermal administration. Another 60 rabbits were divided into several groups as above except for a normal group. A single pri-mary skin irritation test was conducted on them,where skin irritation reactions were recorded 6 h after a single administration based on intra-individual left/right self comparison method. 70 guinea pigs were randomized into a negative control group (vase-line),a PIB group,a positive control group(2,4-dinitrochlorobenzene),a Gutong plaster group and the groups of low,medium and high-dose PIB Gutong plaster,which were dosed for sensitization,followed by a skin sensitization test. RESULTS:No obvi-ous toxicity symptoms could be seen after administration of PIB Gutong plaster. The rabbits’intact or damaged skin had no irrita-tion response to PIB and low and medium-dose PIB Gutong plaster. PIB Gutong plaster caused no irritation response in the rabbits’ intact skin,but slight irritation in damaged skin 1 h after administration. The allergic reaction incidence of the positive control group of guinea pigs was 100% while that of any other groups was 0. CONCLUSIONS:The PIB Gutong plaster is safe for trans-dermal administration.

Antimicrobial and cytotoxic effects of phosphoric acid solution compared to other root canal irrigants

Phosphoric acid has been suggested as an irrigant due to its effectiveness in removing the smear layer. Objectives : The purpose of this study was to compare the antimicrobial and cytotoxic effects of a 37% phosphoric acid solution to other irrigants commonly used in endodontics. Material and Methods : The substances 37% phosphoric acid, 17% EDTA, 10% citric acid, 2% chlorhexidine (solution and gel), and 5.25% NaOCl were evaluated. The antimicrobial activity was tested against Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Actinomyces meyeri, Parvimonas micra, Porphyromonas gingivalis, and Prevotella nigrescens according to the agar diffusion method. The cytotoxicity of the irrigants was determined by using the MTT assay. Results : Phosphoric acid presented higher antimicrobial activity compared to the other tested irrigants. With regard to the cell viability, this solution showed results similar to those with 5.25% NaOCl and 2% chlorhexidine (gel and solution), whereas 17% EDTA and 10% citric acid showed higher cell viability compared to other irrigants. Conclusion : Phosphoric acid demonstrated higher antimicrobial activity and cytotoxicity similar to that of 5.25% NaOCl and 2% chlorhexidine (gel and solution). .

A 14-Day Repeated Dose Toxicity of Epimedii Herba Aqueous Extract Administered by Oral Gavage in F344 Rats

The objective of this study is to characterize a toxicity of Epimedii Herba (EH) in F344 rats and to find a dose levels for the 13 weeks toxicity study. EH is well known as medicinal herb in many Asian countries for traditional medicines of antibacterial and antiviral effects, estrogenic and antiestrogenic effects, and for treatment of osteoporosis, hypotensives, fatigue, kidney disorders, and related complications. However, the indispensable and basic information of toxicological evaluation of EH extract is insufficient to support its safe use. Therefore, we conducted toxicological evaluation of this drug in compliance with OECD and MFDS guideline in this study. The extract of EH was administered orally to F344 rats at dose levels of 0, 500, 1000, 2000, 3500, and 5000 mg/kg/day for 2 weeks. Each group was composed of 5 male and female rats. In this study, there were no treatment of EH-related adverse changes in clinical observations, mortality, body weights, food consumption, urinalysis, gross finding at necropsy, and organ weight examination. Total red blood cell count, hematocrit, mean corpuscular hemoglobin concentration, total cholesterol, and phospholipid were decreased in males and females at 5000 mg/kg/day compared to the control animals. Mean corpuscular volume and reticulocyte counts were increased in males and females at 5000 mg/kg/day compared to control animals. Therefore, we recommend that dose level of 5000 mg/kg/day is a highest treatment group in 13-week EH extract exposure study for further toxicity assessment.

Safety Evaluation of Methoxyflavones Mixture (Ⅰ)from <i>Kempferia parviflora</i>; 28-day Repeated Dose Oral Toxicity Test and Mouse Micronucleus Test / 日本補完代替医療学会誌

Kaempferia parviflora rhizome is used in traditional folk medicines for the treatments of various symptoms in Thailand since ancient times. Several types of methoxyflavones were identified from that plant and the functions of some of those were reported. We determined that five kinds of methoxyflavones (5-hydroxy-3,7,3’,4’-tetramethoxyflavone, 5-hydroxy-7-methoxyflavone, 5-hydroxy-3,7-dimethoxyflavone, 5-hydroxy-3,7,4’-trimethoxyflavone, 5-hydroxy-7,4’-dimethoxyflavone) were included the following treatments of K. parviflora rhizome. The 80 %ethanol extract of that were adsorbed resin, removed 70 % ethanol elution and the rest adsorbed materials were eluted with 99.5 % ethanol. The safety of that five methoxyflavones mixture was evaluated. We performed a 28-day repeated dose of oral toxicity test and a mouse micronucleus test. The former results showed no significant toxicity on body weight, blood analyses, organ weight, blood biochemical analyses. The latter results showed negative, believed that the sample has no mutagenicity for living bodies.

Acute Intravenous Injection Toxicity Study of Escherichia coli-Derived Recombinant Human Bone Morphogenetic Protein-2 in Rat

STUDY DESIGN: Prospective in vivo toxicity study. PURPOSE: To evaluate the conducted acute toxicity study of Escherichia coli (E. coli)-derived recombinant human bone morphogenetic protein-2 (rhBMP-2) with 6-weeks old Sprague-Dawley rats. OVERVIEW OF LITERATURE: rhBMP-2 has well-known osteoinductivity and it is used as a bone graft substitute. E. coli-derived rhBMP-2 can be mass-produced with relatively low costs. E. coli-derived rhBMP-2 facilitates osteoblastic differentiation and bone formation in vitro and in vivo. However, studies regarding side effects or toxicity of E. coli-derived rhBMP-2 have not been published. Thus, we conducted the acute toxicity study of E. coli-derived rhBMP-2 on 6-weeks old Sprague-Dawley rats. METHODS: One mg of BMP-2 was diluted in 0.285 mL of glycine buffer to prepare high BMP-2 concentrations (3.5 mg/mL). Intermediate (0.9 mg/mL) or low (0.35 mg/mL) concentrations of BMP-2 solution was prepared by serial dilutions. The compound was administrated at a dose of 0, 0.7, 1.8, 7 mg/kg by single intravenous injection to five of male and female rats. After the injection, the gross general observations including changes of body weight and histopathological analysis was performed for 14 days. RESULTS: No animal was found dead during the experiment and the body weight changes were both statistically insignificant in the control and experimental groups. No abnormal sign was shown in general observations and autopsy examinations. CONCLUSIONS: Thus, the lethal dose of E. coli-derived rhBMP-2 should be higher than 7 mg/kg with a single intravenous injection.