OBJECTIVES: Ozone is widely applied to treat allergic skin diseases such as eczema, atopic dermatitis, and contact dermatitis. However, the specific mechanism remains unclear. This study aims to investigate the effects of ozonated oil on treating 2,4-dinitrochlorobenzene (DNCB)-induced allergic contact dermatitis (ACD) and the underling mechanisms. METHODS: Besides the blank control (Ctrl) group, all other mice were treated with DNCB to establish an ACD-like mouse model and were randomized into following groups: a model group, a basal oil group, an ozonated oil group, a FcεRI-overexpressed plasmid (FcεRI-OE) group, and a FcεRI empty plasmid (FcεRI-NC) group. The basal oil group and the ozonated oil group were treated with basal oil and ozonated oil, respectively. The FcεRI-OE group and the FcεRI-NC group were intradermally injected 25 µg FcεRI overexpression plasmid and 25 µg FcεRI empty plasmid when treating with ozonated oil, respectively. We recorded skin lesions daily and used reflectance confocal microscope (RCM) to evaluate thickness and inflammatory changes of skin lesions. Hematoxylin-eosin (HE) staining, real-time PCR, RNA-sequencing (RNA-seq), and immunohistochemistry were performed to detct and analyze the skin lesions. RESULTS: Ozonated oil significantly alleviated DNCB-induced ACD-like dermatitis and reduced the expressions of IFN-γ, IL-17A, IL-1β, TNF-α, and other related inflammatory factors (all P<0.05). RNA-seq analysis revealed that ozonated oil significantly inhibited the activation of the DNCB-induced FcεRI/Syk signaling pathway, confirmed by real-time PCR and immunohistochemistry (all P<0.05). Compared with the ozonated oil group and the FcεRI-NC group, the mRNA expression levels of IFN-γ, IL-17A, IL-1β, IL-6, TNF-α, and other inflammatory genes in the FcεRI-OE group were significantly increased (all P<0.05), and the mRNA and protein expression levels of FcεRI and Syk were significantly elevated in the FcεRI-OE group as well (all P<0.05). CONCLUSIONS Ozonated oil significantly improves ACD-like dermatitis and alleviated DNCB-induced ACD-like dermatitis via inhibiting the FcεRI/Syk signaling pathway.
Animals ; Mice ; Dinitrochlorobenzene/metabolism* ; Skin/metabolism* ; Cytokines/metabolism* ; Interleukin-17/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Dermatitis, Allergic Contact/pathology* ; Dermatitis, Atopic/chemically induced* ; Signal Transduction ; RNA, Messenger/metabolism* ; Mice, Inbred BALB C

Objective: To study the effects of Lycium barbarum polysaccharides ( LBP ) on blood indexes and liver tissue morphology in rats with intrahepatic cholestasis. Methods: Sprague-Dawley rats were randomly divided into the control group, the model group, and LBP low, medium and high dose group. The rats in the model group and LBP dose groups were given 60 mg/kg alpha-naphthylisothiocyanate ( ANIT ) by gavage every three days of the experiment, and the rats in the control group were given salad oil instead of ANIT. From the third day, the rats in each dose group were given 40, 150 and 600 mg/kg LBP, and the rats in the model group were given distilled water. After four weeks, the blood and urine indexes were measured, and the morphological changes of liver tissue were observed. Results: From the third day of the experiment, the activity of rats in the model group and LBP dose groups decreased, and the color of urine changed to dark yellow. There was no abnormality in the group. In the model group, the levels of serum total bilirubin, direct bilirubin, total bile acid ( TBA ), alkaline phosphatase ( ALP ), γ-glutamyltransferase(γ-GGT), cholesterol, alanine aminotransferase ( ALT ), aspartate aminotransferase ( AST ), white blood cell ( WBC ), percentage of granulocyte, urinary bilirubin, urinary bile acid, liver mass and liver to body ratio were higher than those in the control group, while red blood cell and percentage of lymphocyte were lower than those in the control group ( all P<0.05 ). Pathological changes of liver tissue were observed. The levels of serum TBA, ALP, γ-GGT, ALT, AST, WBC and liver to body ratio in LBP high dose group were lower than those in the model group ( all P<0.05 ). The infiltration of inflammatory cells, proliferation and expansion of bile duct, degeneration and necrosis of liver cells were alleviated. Conclusions LBP can improve the blood indexes and pathological changes of liver tissue in rats with intrahepatic cholestasis at the dosage of 600 mg/kg. Inhibition of inflammatory response and reduction of oxidative stress injury may be the mechanism for alleviating cholestatic liver injury.

Objective: To learn the effects of Dendrobium officinale flowers on testivular tissue morphology and sperm quality in parent and offspring rats,so as to provide reference for safety evaluation of Dendrobium officinale flowers. Methods: The 40 SD rats was randomly divided into the low-,middle-,high-dose and the control group,given 2.0,4.0,6.4 and 0 g/kgbw Dendrobium officinale flowers,respectively. After three months,the body weight,mass and organ/body coefficients of testis and epididymis of parent （P） and offspring （F1,F2） rats were measured;the number,activity and deformity of sperms were observed under microscope;the changes of testis and epididymis were observed by hematoxylin eosin staining. Results: There were no significantly statistical differences in the body weight,mass and organ/body coefficients of testis and epididymis,sperm quantity,sperm motility rate among four groups of P、F1、F2 male rats （P>0.05）. There were no significantly statistical differences in sperm malformation rate between the high-dose group and the control group （P>0.05）. There was no significant change in testis and epididymis of P,F1 and F2 male rats. Conclusion Dendrobium officinale flowers did not show obviously adverse effects on testivular tissue morphology and sperm quality in parent and offsping rats.

Objective: To learn the toxicity of Dendrobium officinale flowers to pregnant rats ( P, F1 ) and offspring rats ( F1, F2 ) before birth, so as to provide toxicological evidence for the safety assessment. Methods : The rats were divided into four groups with 20 female rats and 10 male each. The rats in three dose groups were fed with Dendrobium officinale flowers at the dose of 2.0, 4.0, 6.4g/kgbw. After two generation, the F1a and F2a rats were fed with basal diet; F1b and F2b rats were fed with Dendrobium officinale flowers. The body weights and total weight gains during the gestation, the conception rates, the pregnancy rates, the birth weights and survival rates of offspring rats were examined. Results: There were no statistically significant differences in the body weights and total weight gains during the gestation, the conception rates, and the pregnancy rates in pregnant rats ( P, F1 ) among the four groups ( P>0.05 ). There were also no statistically significant differences in the survival rates and live birth rates in offspring rats (F1, F2) between the dose groups and the control group ( P>0.05 ). Conclusions Dendrobium officinale flowers did not show obviously adverse effects on pregnant rats ( P, F1 ) and offspring rats ( F1, F2 ) before birth.

Objective: To explore the cause and type of pigmentation in the livers and kidneys of rats caused by a compound Chinese medicine preparation. Methods: The experiment consist of low, medium, and high dose groups and a control group, the Sprague-Dawley rats in these groups were orally given 1.0, 2.0 and 4.0 g/kgbw of a compound Chinese medicine preparation of gardenia and distilled water for 30 days, respectively. The body weight, diet, hematology and histopathology of the rats in each group were observed for changes in pigment metabolism. Results: In the first and second weeks of the experiment, the rats in the low, medium, high dose groups and the control group showed no abnormal symptoms or signs. From the third weekend, the urine of the rats in the high dose group turned thick yellow and green, and the stool color became light. During the experiment, no rats died. There were statistically significant differences in body weights and weight gains among these groups ( P<0.05 ) . There were statistically significant differences in total food utilization, direct bilirubin, r-glutamyl transferase, alkaline phosphatase, and liver/body ratios among these groups ( P<0.05 ). Gross examination revealed that the livers and kidneys of rats in the high dose group were dull and green. Microscopic examination revealed changes in dark pigment particles in the livers and kidneys of rats in the high dose group. Histochemical staining confirmed that pigments in the livers and kidneys were bile pigments. Conclusions At a dosage of 4.0 g/kgbw, a compound Chinese medicine preparation of gardenia can lead to bile pigment deposition in the livers and kidneys of rats due to cholestasis.

Objective: To evaluate the toxicological safety of a compound Chinese medicine preparation of gardenia. Methods: Eighty healthy SD rats with half males and half females were randomly divided into four groups. The low-,moderate- and high-dose group were given 1.00 g/kgbw,2.00 g/kgbw and 4.00 g/kgbw of the preparation,while the control group was given distilled water,by gavage for 30 days. The changes of diet,weight,hematological parameters and major organs of rats were observed,and the histopathological examination of the main organs was performed. Results: The rats in the high-dose group reduced activities and their urine turned dark yellow or green,while the other rats showed no abnormality. No rats died during the experimental period. Compared with the control group,the weight,the total weight gain,the food utilization rate,the fasted weight of the rats in the high-dose group and the hemoglobin content of the female rats in the high-dose group were significantly decreased（P<0.05）;the ratio of liver to body weight,the ratio of kidney to body weight,the serum creatinine levels of the rats in the high-dose group and the serum urea nitrogen levels of the male rats in the high-dose group were significantly higher（P<0.05）. The livers and kidneys of the rats in high-dose group turned different degrees of dark green;the hepatic pigmentation,hepatocyte vacuolar degeneration,bile duct hyperplasia accompanied with inflammatory cell infiltration,renal pigmentation,renal tubular epithelial cellular swelling,and renal interstitial inflammatory cell infiltration were observed. Conclusion This preparation at a dose of 4.0 g/kgbw has hepatotoxicity and nephrotoxicity to rats. A dose of 2.0 g/kgbw has no harmful effect but less than 100 times of the possible human intake,the safety is not guaranteed.

Objective: To evaluate the safety of a herbal pilatory for external use. Methods : An acute eye irritation test were employed to detect the eye irritation of the herbal pilatory;a skin irritation test,a skin sensitization test and a skin phototoxicity test were employed to detect the dermal toxicity;Salmonella typhimurium reverse mutation assay（Ames test）and chromosome aberration test in CHL cells were employed to detect the effects of the pilatory on gene mutation and chromosome aberration in prokaryotic and eukaryotic cells. Results: When the eyes of rabbits exposed to the pilatory without rinse during the first 24 hours,the conjunctiva showed congestion and edema with the highest score of 2,corneal opacity was observed with the highest score of 1;however,these symptoms returned to normal within 72 hours,with the score reduced to 0. No irritation to the skin of rabits was found after exposed to the pilatory for fourteen days,no skin sensitization was introduced by Buehler test and no skin phototoxicity on guinea pigs was detected. There was no abnormal growth of reverse mutation colonies induced by the pilatory under S9 acitivation or not. There was no statistically significant rise of chromosome aberration rate in the exposed CHL cells compared to the control groups（P>0.05）. Conclusion Under the condition,the herbal pilatory showed mild and reversible irritation to eyes,while no dermal toxicity and genetic toxicity were observed. The safety of the herbal pilatory for external use is acceptable.

Objective : To evaluate the health effects of persistent organic pollutants（POPs）on body weight,food intake,internal organs,blood biochemistry,metabolic enzymes and antioxidant ingredients of rats. Methods : Sixteen healthy Sprague-Dawley（SD）rats were randomly divided into the experimental group exposed to 10 mL/kg mixture of POPs（10 ng/mL PCBs,5 ng/mL PBDEs,1 ng/mL PCDD/F）everyday for 28 days by gavage,and the control group exposed to the same volume of soybean oil in the same way. Body weight and food intake of the rats were recorded regularly;blood routine and biochemical indices were detected;liver,kidney,spleen and testicles（ovary）of the rats were weighed to calculate organ coefficients;metabolic enzymes and antioxidant ingredients were detected from livers of the rats. Results : No obviously abnormal symptoms and no deaths were found in both groups. Compared to the control group,the weekly food intake in the experimental group increased more for there was an interaction between grouping and time（P< 0.05）. The ratio of liver to body weight of male rats in the experimental group was higher than that in the control group [（3.87± 0.19）% vs.（3.53± 0.06）%,P< 0.05]. The haemoglobin and red blood cell of female rats in the experimental group were lower than those in the control group[（145.25± 6.18）g/L vs.（154.50± 4.20）g/L;（6.90± 0.14）× 1012/L vs.（7.39± 0.24）× 1012/L;both P< 0.05]. The glutathione-S-transferase（GST）of female rats in the experimental group was higher than that in the control group [（13.37± 1.05）U/mgprot vs.（9.43± 1.08）U/mgprot,P< 0.05]. The cytochrome P4501A1of rats in the experimental group was higher than that in the control group [female：（88.23± 5.81）ng/mgprot vs.（73.85± 5.86）ng/mgprot;male：（96.80± 13.32）ng/mgprot vs.（ 72.20± 2.01）ng/mgprot;both P< 0.05]. Conclusion After exposed to low dose of POPs,the cytochrome P4501A1 increased in all rats,the liver to body weight ratio increased in male rats,GST activity increased while red blood cell and haemoglobin decreased in female rats,which indicated possible body damages in rats.

Objective:To evaluate skin irritation,acute toxicity,and allergy of medical ozone oil for clinical application.Methods:In contrast to their left and right side irritation,one or more skin irritation tests were performed on the intact and damaged skins of guinea pigs.With the maximum concentration,acute skin toxicity test was applied on the intact and damaged skins of rats.Active cutaneous anaphylaxis was applied to the guinea pigs.Results:High concentration (ozone consumption:150 g/L) of medical ozone oil showed a slight irritation on the broken skin of guinea pigs,while low concentrations did not show skin irritation.Medical ozone oil had no obvious acute toxicity to rats.The medical ozone oil and base oil showed mildallergy for the skin of guinea pig.Conclusion:The irritation of medical ozone oil is related to its concentration.With appropriateconcentration and duration of treatment,medical ozone oil is safe.

Objective:To explore a new method for detecting the bactericidal effect of oiling agent in vitro,and to determine the disinfectant effecacy ofozonated camellia oil on Staphylococcus aureus.Methods:Suspension of Staphylococcus aureus was prepared and innoculated on the LB plate by plate scribing method.After culture overnight,21 bacterial monoclones with the same diameter were selected and divided into 3 groups:A negative control group,a baseoil (camellia oil) group and an ozonated camellia oil group.We used a ring to isolate the single clone and added oil inside the ring,cultured the whole plate over night,picked out each single clone (with gel) to 5 mL LB medium and cultured it for 12 h.The final concentration of the LB medium was detected by plate count method and turbidimetry.Results:According to the plate count method and turbidimetry,the bacterial concentration in the ozonated camellia oil group was lower than that in the negative control group and base oil group Conclusion:Bacterial monoclone culture method shows that ozonated camellia oil can significantly kill Staphylococcus aureus,and this method is an effective method for evaluating the bactericidal function of the oiling agent in vitro.