Bladder calculi is a type of urinary calculi,which is mostly due to geographical environ-ments,dietary habits,ethnic genetics and diseases of the urinary tract.It is necessary to establish or improve animal models of bladder calculi in order to research the mechanism and develop drugs for the disease.There are 4 types of construction methods for bladder calculi models,which are physical induction,chemical induction,microbial induction and gene knockout models.This review classifies and summarizes the construction methods,characteristics,advantages and disadvantages of rodent ani-mal models for bladder calculi,focusing on the construction processes,cycles and effectiveness.Differ-ent models were compared and problems with the application of present models in drug research and development were pointed out in order to provide reference for the non-clinical effectiveness and safety studies of drugs and devices for the prevention and treatment of bladder calculi.

OBJECTIVE To study the changes in inflammatory factors caused by prostatitis.METHODS SD rats were castrated under sterile conditions.E2 0.25 mg· kg-1+ T 0.25,0.5 and 1.0 mg· kg-1 were given sc for 30 d,respectively.Serum samples were taken and levels of E2,T and dihydrotestosterone (DHT) were detected by ELISA.Pathological changes of prostate tissue were observed by HE staining.The expressions of tumor necrosis factor-alpha (TNF-α),cyclooxygenase-2 (COX-2) and macrophage inflammatory protein 1alpha (MIP-1α) in prostate were detected by immunohistochemistry.RESLULTS ELISA detection showed that E2 levels were significantly increased [(80±7) ng· L-1,P＜0.01] in E2 0.25 mg· kg-1 group and that T levels were significantly decreased [111 ±6 vs (111 ±5) nmol· L-1,P＜0.05]in E20.25 mg ·kg-1 and E2+T 0.25 mg·kg-1 groups compared with the sham-operated group.E2 was significantly increased [(80±7) ng· L-1,P＜0.01] in E20.25 mg· kg-1 groups compared with the castrated control.The sham and castrated control showed normal glandular epithelium without leukocyte infiltration.In E2 0.25 mg·kg-1 group,extensive infiltration of inflammatory cells was found in the glandular lumens,suggesting the occurrence of chronic prostatitis.In each E2+T groups,fewer inflammatory cells were noted in the stroma around glands.The expressions of TNF-m COX-2 and MIP-1α in sham group were negative or low,while those of castrated control and E2 0.25 mg· kg-1 groups were high,especially in E2 0.25 mg· kg-1 group.The expressions of TNF-α,COX-2 and MIP-1α in each E2+T group were significantly decreased.CONCLUSION Testosterone can inhibit prostatitis and the expression of inflammatory factors,such as TNF-α,COX-2 and MIP-1 α,in castrated SD rats initiated by estrogen.

OBJECTIVE To investigate the proliferation effect of di(2-ethylhexyl) phthalate (DEHP) on prostate in aged rats at the environmental exposure dose and the possible mechanism.METHODS Thirty-two male Sprague-Dawley rats,aged 1.5 years,were randomly divided into 4 groups (8 rats per group) and treated with DEHP (30,90 and 270 μg· kg-1,ig) and vehicle once daily respectively for 4 weeks.All the animals were anesthetized with pentobarbital sodium and sacrificed on the day subsequent to the last treatment.① Abdominal aortic blood samples were collected,and serum estradiol (E2),testosterone (T) and prolactin (PRL) levels were assayed by ELISA.② The prostate tissues were dissected and categorized into different lobes,weighed and measured.The prostate relative mass was calculated.③ The morphological changes were detected by HE staining and prostate epithelial height was analyzed with microscopic image analysis software.RESULTS Compared with vehicle control group,the prostate relative mass,dorsolateral prostate mass,and dorsolateral prostate index in DEHP 270 μg· kg-1 group were significantly higher (P＜0.05).The height of the ventral prostate epithelium in DEHP 30,90 and 270 μg· kg-1 groups was increased significantly (P＜0.01),so was the height of dorsal prostate epithelium in DEHP 270 μg· kg-1 group (P＜0.01).There were no significant changes in levels of E2,PRL or T in DEHP 30,90 and 270 μg· kg-1 groups,but the ratios of E2/T in DEHP 30 and 270 μμg· kg-1 groups were increased significantly (P＜0.05).CONCLUSION Low-dose DEHP could promote the proliferation of prostatic hyperplasia in the aged rats,which might be associated with the relative levels of endogenous hormone.

Prostate cancer is the most common cancer among males in the United states,and the incidence has risen dramatically in recent years in China. Lymph node metastasis is a strong predictor of the metastatic potential and poor outcome of prostate cancer. Animal models of human prostate cancer lymphatic metastasis can be used to study the pathogenesis and metastatic mechanisms of prostate cancer,and evaluate the efficacy of new drugs for lymphatic metastasis of prostate cancer. This paper reviews commonly-used animal models of human prostate cancer lymphatic metastasis,including xenograft mouse models,genetically engineered mouse models,rat models and canine models,analyzes their advantages and disadvantages,presents their functions and characteristics,introduces the applications of cancer stem cells in these models and test methods of these models,and highlights the main problems to be solved.

In current research and development of new drugs,the demand for toxicological study using neonatal and juvenile animals is becoming increasingly urgent. In this paper,we discussed the characteristics,importance and necessity of nonclinical safety evaluation for pediatric drugs,considerations for research design,selection of animal species and age,route and duration of drug administration and evaluation indexes. In addition,the characteristics of nonclinical safety evaluation of new traditional Chinese materia medica used for children were analyzed. It is hoped that these studies will not only provide support and reference for nonclinical safety evaluation of pediatric drugs but help accumulate material in formulating relevant guidelines.

There is an increasing demand for neonatal and juvenile animal toxicity studies during the research and development of new drugs. In this paper,we discussed general evaluation parameters of pediatric non-clinical safety with pediatric drugs,such as growth and development and food intake,and paramenters of other organs and systems, such as the central nervous system,reproductive system, behavior evaluation in combination with our own experience. In addition,the characteristics of non-clin?ical safety evaluation of new traditional Chinese medicine materia medica used for juvenile animals were analyzed. This paper is intended reference for non-clinical safety evaluation of pediatric drugs and to gain some experience related to formulation of new guidelines.

OBJECTIVE To compare the difference of methods for active systemic anaphylactic reaction induced by breviscapine injection between ″Pharmacopoeia″ 2010 edition, an Attached ⅫG in Traditional Chinese Medicine lnjection Safety Test Application Guidelines ″ Anaphylactic Reaction Test″(thereafter referred to as the method of ″Pharmacopoeia″) and the Traditional Chinese Medicine, Natural Medicine lmmune Toxicity (Anaphylaxis, Anaphylactic Reaction of Light) Technology Guiding Principles in the 2005 Version (thereafter referred to as the method of ″Guiding Principle″) and provide reference for non-clinical safety evaluation of drugs. METHODS According to the methods of ″Pharmacopoeia″and ″Guiding Principle″, respectively, the effect of breviscapine injection on active systemic anaphylactic reaction of guinea pigs was investigated. The guinea pigs were divided into four groups, negative control group, positive control group, breviscapine injection 5 and 50 mg.kg-1 groups. ln the sensitization phase, the guinea pigs were ip administrered with breviscapine injection 0.5 mL each every other day for 3 times. The dose was 5 and 50 mg.kg-1 , respectively. For the method of ″Pharmacopoeia″, on the 14th and 21st days after the first sensitization, the guinea pigs were iv administrered with breviscapine injec-tion 1 mL. For the method of ″Guiding Principle″, the guinea pigs were provocated on the 12th day after the first sensitization. Each group was studied by observing the symptom of anaphylactic reaction and immune time. RESULTS For the method of ″Pharmacopoeia″, on the 14th day after the first sensitization, there was 1 guinea pig with sneezing and (or) the nose-scratching at different time in the 5 mg.kg-1 group. ln the 50 mg.kg-1 group, there was one or two cases of sneezing and (or) 1 case of nose-scratc-hing. The 5 and 50 mg.kg-1 dose groups conformed to the regulations. On the 21st day after the first sensitization, trembling occurred in the 5 mg.kg-1 group, with 1 or 2 guinea pigs sneezing and ( or) scratching the nose. There were 4 guinea pigs (4/ 4) with sneezing 1 and 3 times, cough once or twice, 1 scratching nose and urination at different time, and 1 guinea pigs (1/ 4) appeared 3 times consecutive sneezing and shivering in 50 mg.kg-1 group. The 5 mg.kg-1 group conformed to the regulations, while the 50 mg.kg-1 group did not. For the method of ″Guiding Principle″, the 5 mg.kg-1 group was weak positive or positive, with different degrees of symptoms of an anaphylactic reaction, including 3 guinea pigs scratched nasal symptoms. And the 50 mg.kg-1 group of anaphylactic symptoms including scratc-hing nose, sneezing, coughing and (or) urination, showed positive. CONCLUSION During the active systemic anaphylactic reaction of drugs non-clinical safety evaluation of drugs the advantage of either method should be brought into play. The method of ″ Pharmacopoeia″ may be used for preliminary screening of test samples. ln case pf suspected reactions, the method of ″Guiding Principle″ should be used for more detailed observations.

Objective To explore the relationships between active systemic allergic reaction induced by breviscapine injection and the drug dose and the sensitization time. Methods Active systemic allergic reaction in guinea pigs was used as experimental method. Forty-eight guinea pigs were divided into 6 groups according to random number table:breviscapine injection 1,5,25 and 50 mg/kg group( the breviscapine injection group 1,2,3,4),0. 9% sodium chloride injection group( the negative control group)and bovine serum albumin( BSA)control group( the positive control group). Each group comprised 8 guinea pigs. Sensitization:the guinea pigs in group 1 to 4 were given the breviscapine injection at doses of 1,5,25 and 50 mg/kg(0. 5 ml)by intraperitoneal injection every other day for three times,respectively. The the guinea pigs in the negative control group and the positive control group were given 0. 9% sodium chloride injection(0. 5 ml)and bovine serum albumin( BSA)20 mg/kg by intraperitoneal injection every Excitation:the sensitized guinea pigs in each group were divided into 2 subgroups,each subgroup comprised 4 guinea pigs. On the 14 and 21 days after the last sensitization,the guinea pigs in breviscapine 1 to 4 subgroups received 2 times of breviscapine injection intravenously(1. 0 ml),respectively. The guinea pigs in the negative and the positive control subgroups received 2 times of control articles intravenously,respectively. The symptoms of anaphylactic reaction( pilo-erection,shiver, scratching nose,sneeze,cough,vomiturition,cyanosis,dyspnea,urinary and fecal incontinence,instability of gait or tumble,convulsion or hyperspasmia,shock and death ) were observed every day during the sensitization phase. The guinea pigs′ reactions which appeared in 30 min after intravenous injections were observed and the occurrence time of allergic symptoms/signs were recorded attentively. The anaphylactic reaction was determined according to the Chinese Pharmacopoeia′s allergic reaction test. Results The guinea pigs in 6 groups did not show any allergic symptoms in the sensitization phase. Within 30 min in fourteenth days′excitation,2,4,4,4 guinea pigs developed allergic reactions in the breviscapine 1 to 4 groups,respectively. But none of them were identified as allergic reaction. The 4 guinea pigs in the positive control group were judged to have positive allergic reaction. Within 30 min of 21 days′excitation,4、3、4、4 guinea pigs developed allergic reactions in the breviscapine 1 to 4 groups,respectively. Only one guinea pig in the breviscapine 4 group was identified as allergic reaction. The 4 guinea pigs in the positive control group were all judged positive allergic reaction. The guinea pigs in the negative control group did not develop any allergic reactions during the 2 times of excitation. The occurrence time of allergic reactions in the breviscapine groups( within 20 min after excitation)on 21 days′excitation was shorter than those( within 25 min after excitation)occurred on 14 days′ excitation,but the difference was not statistically significant. Conclusions The active systemic allergic reactions induced by breviscapine injection are associated with drug dose and sensitization time. The larger dose and longer sensitization time can increase the risk of allergic reaction and shorten the latency of allergic reaction.

Objective To explore the relationships between active systemic allergic reaction induced by breviscapine injection and the drug dose and the sensitization time. Methods Active systemic allergic reaction in guinea pigs was used as experimental method. Forty-eight guinea pigs were divided into 6 groups according to random number table:breviscapine injection 1,5,25 and 50 mg/kg group( the breviscapine injection group 1,2,3,4),0. 9% sodium chloride injection group( the negative control group)and bovine serum albumin( BSA)control group( the positive control group). Each group comprised 8 guinea pigs. Sensitization:the guinea pigs in group 1 to 4 were given the breviscapine injection at doses of 1,5,25 and 50 mg/kg(0. 5 ml)by intraperitoneal injection every other day for three times,respectively. The the guinea pigs in the negative control group and the positive control group were given 0. 9% sodium chloride injection(0. 5 ml)and bovine serum albumin( BSA)20 mg/kg by intraperitoneal injection every Excitation:the sensitized guinea pigs in each group were divided into 2 subgroups,each subgroup comprised 4 guinea pigs. On the 14 and 21 days after the last sensitization,the guinea pigs in breviscapine 1 to 4 subgroups received 2 times of breviscapine injection intravenously(1. 0 ml),respectively. The guinea pigs in the negative and the positive control subgroups received 2 times of control articles intravenously,respectively. The symptoms of anaphylactic reaction( pilo-erection,shiver, scratching nose,sneeze,cough,vomiturition,cyanosis,dyspnea,urinary and fecal incontinence,instability of gait or tumble,convulsion or hyperspasmia,shock and death ) were observed every day during the sensitization phase. The guinea pigs′ reactions which appeared in 30 min after intravenous injections were observed and the occurrence time of allergic symptoms/signs were recorded attentively. The anaphylactic reaction was determined according to the Chinese Pharmacopoeia′s allergic reaction test. Results The guinea pigs in 6 groups did not show any allergic symptoms in the sensitization phase. Within 30 min in fourteenth days′excitation,2,4,4,4 guinea pigs developed allergic reactions in the breviscapine 1 to 4 groups,respectively. But none of them were identified as allergic reaction. The 4 guinea pigs in the positive control group were judged to have positive allergic reaction. Within 30 min of 21 days′excitation,4、3、4、4 guinea pigs developed allergic reactions in the breviscapine 1 to 4 groups,respectively. Only one guinea pig in the breviscapine 4 group was identified as allergic reaction. The 4 guinea pigs in the positive control group were all judged positive allergic reaction. The guinea pigs in the negative control group did not develop any allergic reactions during the 2 times of excitation. The occurrence time of allergic reactions in the breviscapine groups( within 20 min after excitation)on 21 days′excitation was shorter than those( within 25 min after excitation)occurred on 14 days′ excitation,but the difference was not statistically significant. Conclusions The active systemic allergic reactions induced by breviscapine injection are associated with drug dose and sensitization time. The larger dose and longer sensitization time can increase the risk of allergic reaction and shorten the latency of allergic reaction.

Elements, including experimental personnel, facili-ties, equipment, apparatus, technology and skills, are essential to carrying out pre-clinical reproductive toxicity research. As to a study , authenticity , normativity , scientificity and innovativeness guarantee the success. These four elements are independent of each other,but mutually supported. Authenticity and scientificity are the footstone and soul, respectively, for drug non-clinical re-productive toxicity research. Normativity guarantees authenticity and scientificity, and innovativeness relies on authenticity. Nor-mativity and scientificity ensure the reliability and dynamics of experimental results which perhaps is an effective way accessing innovation.