Andrographis paniculata as Promising Novel Protective Therapy of Oxidative Stress in Indomethacin-Induced Gastric Ulcer in Rats

This research investigated the effect of Andrographis paniculata (AP) on oxidative stress following indomethacin-induced gastric ulcer in rats. A total of 20 male albino Wistar rats (150-180g) used for this study were grouped into four (n=5): 1, Negative Control; 2, Positive Control and 3, test group treated with normal chow, 20mg/kg indomethacin, 20 mg/kg indomethacin plus omeprazole at 20mg/kg and 20mg/kg indomethacin plus AP at 16.7 mg/kg respectively. After treatment period, estimation of oxidative stress parameters was carried out on the animals. The LD50 of aqueous extract of AP was 50mg/kg bw. Body weight change was significantly reduced in omeprazole treated group compared to all other groups while extract treated group had significantly increased body weight change. There was a significant increase in malondialdehyde (MDA) level of ulcer untreated group compared to other groups. The two treated groups had significantly reduced MDA compared to ulcer untreated group. There was a significant decrease in the levels of GPx and SOD of ulcer untreated group compared to control. Meanwhile, these were significantly increased in extract and omeprazole treated groups compared to ulcer untreated group. Catalase was significantly increased in all three groups when compared to control but its level was significantly increased in extract treated group compared to ulcer untreated and omeprazole treated groups. From this study, AP has proved to protect against oxidative stress implicated in the pathogenesis of ulcer. If this result is applicable to humans, further research and use of AP in ameliorating debilitating consequences of peptic ulcer should be encouraged.

A Report on the Distribution of "Lethal Dose/Pharmaceutical Product Strength” in High-Risk Drugs / 医薬品情報学

Objective: This study assessed the distribution of "lethal dose/pharmaceutical product strength" in high-risk drugs.Methods: In 707 pharmaceutical products (312 ingredients) that had been defined as high-risk drugs in Japan, we collected acute toxicity information from these products on single dose toxicity studies conducted in mice, including median lethal dose (LD50) and approximate lethal dose (aLD). The LD50 and aLD were then divided by the strength (quantity of active ingredients) of the pharmaceutical product, after which the LD50or aLD values having an inequality sign was excluded.Results: We collected data on the acute lethal dose of 707 products (312 ingredients) from high-risk drugs. Data with an inequality sign, which was 143 of 495 products (28.9%) in tablets and capsules, then 43 of 212 items (20.3%) in injections, were excluded from the analysis. As observed, median (Q1, Q3) of "LD50/pharmaceutical product strength" and "aLD/pharmaceutical product strength" for tablets or capsules was 36.8 tablet/kg (11.5 tablet/kg, 144 tablet/kg) and 16.7 tablet/kg (6.9 tablet/kg, 65 tablet/kg), respectively. However, median (Q1, Q3) of "LD50/pharmaceutical product strength" and "aLD/pharmaceutical product strength" for injections were 1.3 bottle/kg (0.6 bottle/kg, 4.7 bottle/kg) and 0.8 bottle/kg (0.4 bottle/kg, 15 bottle/kg), respectively. In both cases, injections were distributed at a lower value than oral products.Conclusion: From this study, the distribution of "lethal dose/pharmaceutical product strength" in high-risk drugs was clarified. This information will therefore help pharmacists assess risks associated with individual pharmaceutical products.

Review on the In Vitro Cytotoxicity Assessment in Accordance to the International Organization for Standardization (ISO)

@#Cytotoxicity is a predominant biological evaluation applied to search for a suitable and non-toxic bioactive compound and to determine the biocompatibility of medical devices-related human body. The broad usage of cytotoxicity tests leads to a robust establishment of cytotoxicity assays with high sensitivity and prompt results. In vitro assays are always prioritized over in vivo due to the reproducible data, reduce numbers of animal used and easily accessible material. Compounds concentration that execute 50% of cell population is determined by calculating the IC50. According to ISO10993, cytotoxicity tests must be performed to determine the biocompatibility of medical devices that has contact with human body. This is crucial to ensure the safety of research and its clinical use. Under the recommendation of ISO10995-Part 5, three categories of tests have been documented; extract elution, direct contact and indirect contact test. Each category plays significant role depending on the nature of experiment and sample used.

Preparation of Highly Water-soluble Platinum Antitumor Drugs and Evaluation of Antitumor Efficacy on Tumor Cell Lines / 中国药学杂志

OBJECTIVE: To design and synthesize a new kind of highly water-soluble platinum antitumor compounds, and then evaluate their cytotoxicity in order to confirm their antitumor efficacy. METHODS: Diamide-diiodide platinum was firstly synthesized from potassium chloroplatinate, which was then reacted with Ag2SO4 to obtain intermediate . Using disodium 2-amino-alkyl malonate or N-substituted amino alkyl malonate as the intermediate Ⅱ, the two intermediates reacted at 1∶1 molar ratio to obtain the target compound III in the presence of acid. RESULTS: A new class of platinum compounds were synthesized, which had much better water solubility than that of the existing three-generation platinum compounds. Their antitumor efficacy was confirmed against a variety of tumor cell lines which was higher than that of carboplatin. IIIg was similar to cisplatin in antitumor efficacy on some tumor cell lines. Some target compounds were effective against cisplatin-resistant cell lines. CONCLUSION: Currently in the clinical trial, the target compound IIIg is a new platinum-base antitumor candidate, which exhibits good water solubility and antitumor efficacy in vitro, and the LD50 based on mice shows its lower toxicity than that of cisplatin and carboplatin in vivo.

Acute and Sub-acute Toxicity Assessment of Euphorbia lateriflora (Schum and Thonn) in Wistar Albino Rats

Aims: The toxicity of ethanol whole plant extract of Euphorbia lateriflora was assessed in Albino Wistar rats. Methodology: The LD50 was at single dose of 5000 mg/kg body weight, the sub-acute dosage of the extract was administered orally at 250 and 500 mg/kg b.w.t twice daily for 7 days and the effect of the extract on liver, kidney, and haematological parameters was assessed and recorded during these periods. Results: The result of the oral acute toxicity study at single high dose of 5000 mg/kg/bwt shows that the LD50 of the extract is greater than 5000 mg/kg/bwt. After 7 days of oral administration, 500 mg/kg/bwt of the extract caused a significant (p<0.05) decrease in the packed cell volume. At 500 mg/kg/bwt, the extract caused a significant (p<0.05) increase in ALP, total protein and albumin and decrease in serum electrolytes (Na+, k+ and Cl-). Histopathological analysis revealed the expansion of fibrous spaces in the liver and thickening of the glomerular basement of the kidney in the group fed with 500 mg/kg/b.w.t of extracts. Conclusion: In conclusion, the dose and time-dependent selective organ toxicity effect of this extract suggested that the extract might be relatively unsafe for consumption at especially high concentrations.

Pathogenicity of a Strain of Streptococcus suis Type 2 to Mice and Swine Isolated from Jingzhou City

Aims: The objective of this study is to identify S. suis type 2 and evaluate the virulence of ZHJ01 strain isolation, and verity the clinical and pathological outcome of a systemic infection caused by one serotype 2 when simultaneously inoculated with ZHJ01 strain. We also want to clarify the epidemiologic, clinical, microbiologic characteristics and the pathogenesis mechanism of S. suis type 2 in Hubei province, China. Study Design: Pigs suspected of being infected with S. suis in Jingzhou regions of Hubei province, China were studied. S. suis type 2 isolation was obtained from the suspicion of diseased pig. The case of S. suis type 2 was detected by the virulence factor amplification based on PCR detection and bacterial isolation, identification in the laboratory. According to the experimental infections of mice and piglets, pathogenicity of this S. suis type 2 isolation to mice and swine was monitored. This study was conducted in the key laboratory of pathogenic microbiology, College of Animal Science of Yangtze University, and Institute of Black Pigs Research, Yangtze University. Methodology: Proper serological typing can be performed using a co-agglutination test. The typical colonies purificated and cultured were inoculated with Glucose, Lactose, Raffinos, Sorbitol, D (+)-sucrose, Trehalose, 6.5%NaCl, D (-)-Salicin, Hippurate, Esculin hydrate, V-p, etc., then the test results were recorded. Detection of virulence factors were performed using PCR amplification and DNA sequencing. S.suis type 2 isolation was inoculated to mice and piglets for the virulence test, and the observation of the clinical signs and pathological changes. Results: The virulence factor of extracellular protein factor (EF) was determined from ZHJ01 strain based on PCR detection. Sequence analysis indicated that the isolate was very similar to nucleotide homology with others SS2 strains from different county or contries, and there was not much variation. LD50 of S. suis type 2 for mice was 2.5 x 107cfu. LD50 of S. suis type 2 for piglets was 3.92 x 109cfu. Conclusion: The results show that Swine S. suis type 2 has a relatively strong pathogenicity to pigs in Hubei province, China. This study can be, in part, sufficient to explain the pathogenicity for ZHJ01 strain in area of Zhijing, Jingzhou city, China, which may provide insights into the pathogenesis SS2 and more valid data to support the development of S. suis vaccine as well as the epidemiological investigation, further monitoring and effective prevention to S. suis.

Study on the expression of Toll-like receptor 4 and its effect on prognosis in fat embolism mice model / 上海交通大学学报（医学版）

Objective · To investigate the effect of Toll-like receptor 4 (TLR4) in the pathological injury in fat embolism mice model. Methods · One hundred and twenty male C57BL/6 mice were randomly divided into 10 groups. One group was set as blank control group, and others were injected separately with 1, 2…9 μL/g of allogeneic perirenal fat via tail vein, respectively. The mortality of each group was counted, median lethal dose (LD50) of fat injection in mice was calculated by Bliss method, and the fat embolism LD50 mice model was established. The TLR4 protein expression in the pulmonary tissue of surviving mice was detected by Western blotting. Sixty male C57BL/6 mice were randomly divided into the control group (the same dose of saline was given via tail vein) and the experimental groups (group 2 h, group 8 h, group 24 h and group 48 h, the LD50 fat was given via tail vein).The TLR4 protein expression at different time after fat injection was detected by Western blotting. The mortality of 20 TLR4 gene-knockout mice (TLR4-/-mice) was recorded and compared with 60 wild-type mice after LD50 fat injection. Results · The LD50 of fat embolism mice model was (3.93±0.78) μL/g.After the injection of 1-7 μL/g fat, the expressions of TLR4 protein in the pulmonary tissue of all seven groups were significantly increased, compared with the control group (all P=0.000). In the fat embolism LD50 mice model, compared with the control group, the expressions of TLR4 protein in group2 h were significantly increased (P=0.005). Then, expression level of TLR4 protein was gradually reduced after 2 h, and there was no significant difference between the control group and group 48 h. The mortality of TLR4-/- mice injected with LD50 fat was lower than that of wild-type mice (P=0.043).Conclusion · TLR4 protein involves in the pathologic process of fat embolism syndrome. The knockout of TLR4 gene can reduce the mortality of fat embolism mice. TLR4 and its correlated non-infectious inflammatory response may be an important molecular mechanism of biochemical injury in fat embolism syndrome. Blocking the activation of TLR4-mediated signaling pathway can significantly improve the prognosis, which provides new basis for the prevention, evaluation and treatment of fat embolism syndrome.

“Spectral toxicity” relationship of acute toxicity of Sophora alopecuroides extracted by different extraction methods to mice / 中草药

Objective: To establish the chemical fingerprint of Sophora alopecuroides extracts based on high performance liquid chromatography (HPLC), and determine the LD50 of different extracts of S. alopecuroides to analyze its “spectrum toxicity” relationship. Methods: A series of extracts were prepared by 75% ethanol reflux (ER), water decoction (WD), 75% ethanol ultrasound (EU) and water ultrasound (WU), and their fingerprints were established to determine the acute toxicity LD50 of different extracts. The relationship between chemical composition and acute toxicity LD50 of S. alopecuroides extracts were studied by means of fingerprint similarity evaluation system. Results: The LD50 of ER, WD, EU, and WU extracts were 38.397, 24.994, 18.536, and 19.957 g/kg, respectively. The ocular lesions of mice viscera were mainly manifested in liver and kidney, and the toxicity of ER extracts was the greatest. The 10 common peaks of S. alopecuroides extracts can be divided into two categories; Peaks 4 and 10, oxymatrine and sophocarpidine were negatively correlated with acute toxicity LD50. Conclusion: The spectral toxicity relationship analysis method of S. alopecuroides was constructed. The unidentified peaks 4, 10 and oxymatrine and sophocarpidine were the main chemical components of the toxicity reaction, which laid a good foundation for clinical application and scientific and rational development of S. alopecuroides.

Study on the expression of Toll-like receptor 4 and its effect on prognosis in fat embolism mice model / 上海交通大学学报（医学版）

Objective • To investigate the effect of Toll-like receptor 4 (TLR4) in the pathological injury in fat embolism mice model. Methods • One hundred and twenty male C57BL/6 mice were randomly divided into 10 groups. One group was set as blank control group, and others were injected separately with 1, 2…9 μL/g of allogeneic perirenal fat via tail vein, respectively. The mortality of each group was counted, median lethal dose (LD50) of fat injection in mice was calculated by Bliss method, and the fat embolism LD50 mice model was established. The TLR4 protein expression in the pulmonary tissue of surviving mice was detected by Western blotting. Sixty male C57BL/6 mice were randomly divided into the control group (the same dose of saline was given via tail vein) and the experimental groups (group 2 h, group 8 h, group 24 h and group 48 h, the LD50 fat was given via tail vein). The TLR4 protein expression at different time after fat injection was detected by Western blotting. The mortality of 20 TLR4 gene-knockout mice (TLR4-/- mice) was recorded and compared with 60 wild-type mice after LD50 fat injection. Results • The LD50 of fat embolism mice model was (3.93±0.78) μL/g. After the injection of 1-7 μL/g fat, the expressions of TLR4 protein in the pulmonary tissue of all seven groups were significantly increased, compared with the control group (all P=0.000). In the fat embolism LD50 mice model, compared with the control group, the expressions of TLR4 protein in group 2 h were significantly increased (P=0.005). Then, expression level of TLR4 protein was gradually reduced after 2 h, and there was no significant difference between the control group and group 48 h. The mortality of TLR4-/- mice injected with LD50 fat was lower than that of wild-type mice (P=0.043). Conclusion • TLR4 protein involves in the pathologic process of fat embolism syndrome. The knockout of TLR4 gene can reduce the mortality of fat embolism mice. TLR4 and its correlated non-infectious inflammatory response may be an important molecular mechanism of biochemical injury in fat embolism syndrome. Blocking the activation of TLR4-mediated signaling pathway can significantly improve the prognosis, which provides new basis for the prevention, evaluation and treatment of fat embolism syndrome.

Study of Acute Toxicity Different Extraction Parts of Aconitum sinomontanum Nakai on Mice / 中国中医药信息杂志

Objective To compare the toxicity of different extraction parts of Aconitum sinomontanum Nakai;To screen out"toxic sections"of Aconitum sinomontanum Nakai; To provide references for further study on toxicity components of Aconitum sinomontanum Nakai. Methods Systematic solvent method was used to extract the 95% ethanol extracts of Aconitum sinomontanum Nakai,and six different extraction fractions(petroleum ether,chloroform, ethyl acetate, butanol, alcohol and water) were obtained. Median lethal dose (LD50) and maximum dose method were used to conduct comparative study on acute toxicity of different extraction parts of Aconitum sinomontanum Nakai. Results Chloroform, water and butanol extractions in LD50of Aconitum sinomontanum Nakai were 89.65, 1805.40 and 24 409.41 mg/kg, and 95% confidence limits were 76.39~108.41, 1521.60~2240.00 and 20 422.54~24 246.95, respectively. The maximum dose of petroleum ether, ethyl acetate and alcohol extractions were 2686.01, 3108.13 and 28 376.21 mg/kg, respectively. Conclusion The maximum toxicity is the extracted section of chloroform, and the minimal toxicity is the extracted section of ethanol.

FlaA gene deletion of LM90 SB2 inducing sheep encephalitis and the influence on virulence / 中国人兽共患病学报

The flagellin of Listeria monocytogenes is encoded by flaA gene;there is no detail report about the influence of flaA gene on the virulence of LM90 SB2.FlaA gene-deleted strain was constructed successfully with recombination technology.The influence of FlaA on LM90 SB2 virulence was evaluated by motility,BF formation,LD50,etc.Results showed that the colony morphology did not change,gene-deleted strain still had good genetic stability,but its growth was slow and the motility was decreased in the environment at 25 ℃,the morphological structure of the mutant BF was looser and incomplete,LD50 was increased from 4.27 × 106 cfu to 1.62 × 107 cfu.Results indicated that the flaA gene affected the flagellar formation of LM90 SB2 significantly,the ability of the BF formation and the virulence of the flaA deleted strain were decreased obviously.

The experimental study of diquat on the half-Lethal dose and pothological injuny of related organs in wistor rats / 中华劳动卫生职业病杂志

Objective@#To explore the acute toxicity of Diquat in mice and to calculate the median lethal dose (LD50) of Diquat to rats and observe the pathological changes of tissues and organs in rats with different concentrations of Diquat.@*Methods@#Diquat solution of 50 mg/kg was prepared freshly with 1 000 mg of Diquat and dilute the solution with water to a total of 20 ml. A total of 99 healthy adult male Wistar rats were randomly divided into part one, part two and control groups. In the first part, 36 rats were randomly divided into 4 groups: 100 mg/kg group, 200 mg/kg group, 300 mg/kg group and 400 mg/kg group, which were treated with 100 mg/kg, 200 mg/kg, 300 mg/kg and 400 mg/kg of Diquat solution by gavage, respectively. The death and symptoms of poisoning after intragastric administration were recorded, and the maximum tolerated dose and absolute lethal dose were measured. In the second part, 54 rats were randomly divided into 6 groups: 200 mg/kg group, 220 mg/kg group, 240 mg/kg group, 260 mg/kg、280 mg/kg group and 300 mg/kg group, whichwere treated with 200 mg/kg, 220 mg/kg, 240 mg/kg, 260 mg/kg, 280 mg/kg and 300 mg/kg of Diquat solution by gavage, respectively. The survival of rats in different concentration of Diquat was observed and the LD50 was calculated by Excel processing the formula of Koch's method. The control group were given equal volume water under the same experimental conditions. And moreover, the lungs, kidneys, hearts, livers, and brain tissues were collected and fixed by formaldehyde, embedded by paraffin, and sectioned for histopathological light microscopy.@*Results@#The maximum tolerated dose was 240 mg/kg and the absolute lethal dose was 300 mg/kg. The LD50 of Diquat for Rats was 280.58 mg/kg. The high-dose group had significantly more organ damage than the low-dose group after diquat poisoning.@*Conclusion@#The determination of the half-lethal dose of diquat, at the same time observed multiple organs damaged in rats after the diquat quickly poisoned. Kidneys, lungs and heart might be the main organ which was heavily damaged. With the extension of observation time, the organ damage of rats exposed to small doses gradually stabilized.

Acute and 13-week subchronic toxicological evaluations of turanose in mice

BACKGROUND/OBJECTIVES: Turanose, α-D-glucosyl-(1→3)-α-D-fructose, is a sucrose isomer which naturally exists in honey. To evaluate toxicity of turanose, acute and subchronic oral toxicity studies were conducted with ICR mice. MATERIALS AND METHODS: For the acute oral toxicity study, turanose was administered as a single oral dose [10 g/kg body weight (b.w.)]. In the subchronic toxicity study, ICR mice were administered 0, 1.75, 3.5, and 7 g/kg b.w. doses of turanose daily for 13 weeks. RESULTS: No signs of acute toxicity, including abnormal behavior, adverse effect, or mortality, were observed over the 14-day study period. In addition, no changes in body weight or food consumption were observed and the median lethal dose (LD₅₀) for oral intake of turanose was determined to be greater than 10 g/kg b.w. General clinical behavior, changes in body weight and food consumption, absolute and relative organ weights, and mortality were not affected in any of the treatment group for 13 weeks. These doses also did not affect the macroscopic pathology, histology, hematology, and blood biochemical analysis of the mice examined. CONCLUSION: No toxicity was observed in the acute and 13-week subchronic oral toxicology studies that were conducted with ICR mice. Furthermore, the no-observed-adverse-effect level is greater than 7 g/kg/day for both male and female ICR mice.

Acute Toxicity Study of Solanumprocumbens Traditional Zhuang Medicine / 医药导报

Objective To evaluate the safety of Solanumprocumbens by studying its acute toxicity to mice. Methods The dosage of 100% death ( Dm) and 100% survive ( Dn) were determined. Five groups were set between the dosage of Dm and Dn in a 1:0.8 ratio, and then were intragastrically administrated once at the dosage of 250,200,160,128,102.4 g·kg-1 respectively.Toxicity and mortality of mice after intragastricly administration of Solanumprocumbens were observed for 14 days continuously. Results After four hours of administration, there were death in each group except the lowest dosage group (102.4 g·kg-1).Number of death of the groups 250,200,160 and 128 g·kg-1 were 10,8,6 and 3 respectively.LD50 of Solanumprocumbens was 153. 02 g · kg-1 , the 95% confidence interval was ( 136. 55, 171. 47 ) g · kg-1 . Conclusion Solanumprocumbens has a certain toxicity.More attention should be payed to its toxicity for clinical rational drug use.

Acute Toxicity Study of Solanumprocumbens Traditional Zhuang Medicine / 医药导报

Objective To evaluate the safety of Solanumprocumbens by studying its acute toxicity to mice. Methods The dosage of 100% death ( Dm) and 100% survive ( Dn) were determined. Five groups were set between the dosage of Dm and Dn in a 1:0.8 ratio, and then were intragastrically administrated once at the dosage of 250,200,160,128,102.4 g·kg-1 respectively.Toxicity and mortality of mice after intragastricly administration of Solanumprocumbens were observed for 14 days continuously. Results After four hours of administration, there were death in each group except the lowest dosage group (102.4 g·kg-1).Number of death of the groups 250,200,160 and 128 g·kg-1 were 10,8,6 and 3 respectively.LD50 of Solanumprocumbens was 153. 02 g · kg-1 , the 95% confidence interval was ( 136. 55, 171. 47 ) g · kg-1 . Conclusion Solanumprocumbens has a certain toxicity.More attention should be payed to its toxicity for clinical rational drug use.

Chemical constituents of kumquat essential oil analyzed by GC-MS and their acute toxicity in mice / 国际药学研究杂志

Objective To analyze the chemical constituents of kumquat essential oil(KEO)and the acute toxicity of KEO in mice. Methods The chemical constituents of KEO were analyzed by GC-MS. After preliminary test, KM mice were randomly divided into control and KEO groups with 20 mice in each group(10 male and 10 female). The doses of KEO were designed with ratio 0.758 between the 100% and 0% mortality amount as 10, 7.60, 5.70, 4.40, 3.30 and 2.50 ml/kg. The acute toxicity was evaluated by examination of the mice for 14 days after single administration of KEO. Results The principal constituents in KEO were α-pinene(0.46%), β-phellandrene(0.11%), β-myrcene(2.34%), D-limonene(95.62%), geranyl acetate(0.10%), β-copaene(0.67%), and γ-elemene (0.13%), which accounted for 99.43% of the total chemical content of KEO. The half-lethal dose(LD50)of KEO in mice by the acute toxicity test was 6.24 ml/kg with the 95% confidence limit of(5.39-7.33)ml/kg in female mice and 5.73 ml/kg with the 95% confidence limit of(4.91-6.79)ml/kg in male mice. The possible toxic target organs were liver, spleen and gastrointestinal tract. Conclusion KEO with the content of D-limonene was obtained from kumquat for the first time, and the toxicity of the KEO was low according to the toxicity classification standard of World Health Organization.

Chemical constituents of kumquat essential oil analyzed by GC-MS and their acute toxicity in mice / 国际药学研究杂志

Objective To analyze the chemical constituents of kumquat essential oil(KEO)and the acute toxicity of KEO in mice. Methods The chemical constituents of KEO were analyzed by GC-MS. After preliminary test,KM mice were randomly divided into control and KEO groups with 20 mice in each group(10 male and 10 female). The doses of KEO were designed with ratio 0.758 be?tween the 100%and 0%mortality amount as 10,7.60,5.70,4.40,3.30 and 2.50 ml/kg. The acute toxicity was evaluated by examina?tion of the mice for 14 days after single administration of KEO. Results The principal constituents in KEO wereα-pinene(0.46%),β-phellandrene(0.11%),β-myrcene(2.34%),D-limonene(95.62%),geranyl acetate(0.10%),β-copaene(0.67%),andγ-elemene (0.13%),which accounted for 99.43%of the total chemical content of KEO. The half-lethal dose(LD50)of KEO in mice by the acute toxicity test was 6.24 ml/kg with the 95%confidence limit of(5.39-7.33)ml/kg in female mice and 5.73 ml/kg with the 95%confi?dence limit of(4.91-6.79)ml/kg in male mice. The possible toxic target organs were liver,spleen and gastrointestinal tract. Conclu?sion KEO with the content of D-limonene was obtained from kumquat for the first time,and the toxicity of the KEO was low accord?ing to the toxicity classification standard of World Health Organization.

Detection and antibiotic sensitivity test of Klebsiella peneumoniae in Ailurus fulgens / 中国人兽共患病学报

To confirm the etiology of a dead case for a 6 year-old female Ailurus fulgens,one strain of the predominant bacteria from pathologic tissues(heart,liver,spleen,lung and other samples) of the dead Ailurus fulgens were examined and isolated.The isolate was named R1 and no other bacteria were isolated.The bacterial etiological examination(morphological characteristics,biochemical characteristics and 16S rDNA gene detection)of R1 showed that it was identifed as K.peneumoniae.Artificial infection to mice about R1 was also conducted in this study.R1 had strong pathogenicity to mice and the LD50 is 6.5 × 104 CFU/mL.Moreover,the clinical and pathological features of the dead mice were consistent with that of the Ailurus fulgens.To find effective therapeutic drugs of curing other Ailurus fulgens,antibiotic sensitivity test of R1 was conducted,and the results revealed that R1 was highly sensitive to cefotaxime et al,moderately sensitive to amikacin and resistant to penicillin.These data showed that K.peneumoniae was bacterial pathogen leading to death of the Ailurus fulgens and it had strong resistance to penicillins,macrolides and virginiamycin and it had broad drug resistance spectrum.However,R1 is sensitive to cephalosporins and aminoglycoside antibiotics.

Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

Background Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. Methods IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice. Results Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg). Conclusions IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.(AU)

Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens. Methods IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice. Results Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg). Conclusions IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.