OBJECTIVETo observe the effect of total flavones of Epimedium leptorrhizum (YYH-C) on osteoporosis in ovariectomized rats.

METHODOvariectomized female rats were randomly divided into the model group, YYH-C lower, middle and high dose (0.7, 1.4, 2.8 g x kg(-1)) groups, the positive drug Bujiale (0.15 mg x kg(-1)) group, and the sham group. The rats were orally ad-ministrated with drugs for three months. Parathyroid hormone (PTH), procollagen I N-terminal peptide (PINP), alkaline phosphatase (ALP), calcium (Ca) and phosphrous (P) in serum were detected. Femur bones and vertebrae bones of left side were collected to determined bone metrological indexes, including bone mineral density (BMD), bone Ca, and bone ash weight/dry weight percentage. Femur bones of right side were collected to for a morphological observation of bone.

RESULTCompared with the sham group, the model group showed significantly higher PTH and ALP content but obviously lower PINP and Ca content. The three YYH-C 3 groups could resist the decrease of PINP. Specifically, low and middle dose groups could remarkably inhibit the increase of PTH, and the high dose group could increase the Ca content in serum, but without significant effect on the rise in ALP. There was no significant difference in P content in serum in each group. BMD, ash weight/dry weight percentage, Ca and P content of the model group were significantly lower than those in the sham group. The high dose YYH-C group could significantly increase BMD. All of the three YYH-C groups could notably increase ash weight/dry weight percentage and Ca, P content in femur bones and vertebrae bones. YYH-C could significantly increase average thickness, area, area percentage of bony trabeculae, cortical bone area percentage of femoral shaft and the number of osteoblasts on the surface of bony trabeculae, and decrease the number of osteoclasts.

CONCLUSIONYYH-C can effectively control the bone mass loss of rats with ovariectomy-induced osteoporosis, prevent the changes in bone microstructure, and inhibit bone absorption, so as to resist high turn-over osteoporosis after ovariectomy. [Key words] total flavones of Epimedium leptorrhizum; ovariectomized rat; osteoporosis

Alkaline Phosphatase ; metabolism ; Animals ; Bone Density ; drug effects ; Calcium ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Epimedium ; chemistry ; Female ; Flavones ; administration & dosage ; Humans ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; Parathyroid Hormone ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To explore the association of Crohn's disease (CD) with T cell immunoglobulin and mucin domain 3 (Tim-3) gene polymorphisms in patients of Zhejiang Han population in China.Methods A total of 308 CD patients and 573 age-and sex-matched healthy controls were enrolled in our study.Two single nucleotide polymorphisms (SNPs) of Tim-3 (rs1036199 and rs10515746) were examined by the improved multiple ligase detection reaction technique (iMLDR).Analyses of linkage disequilibrium and haplotype were also performed by Haploview 4.2 software in all study subjects.Results In general,the allele and genotype frequencies of Tim-3 (rs1036199 and rs10515746) were not statistically different between CD patients and the controls (all P ＞0.05).According to the Montreal Classification,CD patients were divided into different subgroups.The variant allele (C) and genotype (AC + CC) of rs1036199 were more frequent in CD patients with penetrating diseases than in the controls (10.4％ vs 1.7％,P =0.002;20.8％ vs 3.5％,P =0.023).Similar conclusions were also drawn for the variant allele (A) and genotype (CA + AA) of rs10515746 in patients with penetrating diseases when compared with the controls (10.4％ vs 2.2％,P =0.000;20.8％ vs 4.2％,P =0.033,respectively).The two SNPs of Tim-3 were in strong linkage disequilibrium (D'=1.0,r2 =0.928).The haplotype (AC) formed by their wild-type alleles (A) and (C) was decreased in patients with penetrating CD compared with the controls (89.6％ vs 98.3％,P =0.000).However,the haplotype (CA) formed by their variant alleles was more frequent in patients with penetrating CD than in the controls (10.4％ vs 1.6％,P =0.000).Conclusions Tim-3 (rs1036199 and rs10515746) variations might be correlated with the enhanced risk of penetrating diseases in CD patients.Furthermore,the haplotype (AC) and (CA) formed by the two SNPs might be a protective and a risky factor for penetrating CD respectively.

OBJECTIVETo compare the sensitivity of Brown Norway rats (BN) with Guinea pigs (GP) as allergen assessment animal models.

METHODBN rats and GP were randomly assigned to 1 control group, 2 Bovine serum albumin group (BSA), respectively. Animals in BSA groups of BN rats and GPs were sensitized by intraperitoneal injection of 0.6% BSA 1 ml on day 1, 3, 5, respectively, and irritated by intravenous injection of 2.4% BSA 1 ml on day 7 and day 14 after the last sensitization, while the same volume of normal saline was given to control group on each time point mentioned above. The allergic reactions were scored within 1 h after each irritation treatment, and the sera of both BN rats and GPs were collected to detect IgE concentration by using ELISA. The sera were also applied for passive cutaneous anaphylaxis test (PCA test) in SD rats.

RESULTNo obvious allergic reactions were observed in BSA group of GPs after each irritation treat, however, the score of allergic response in BSA group of BN rats was evidently higher than that in control group after first irritation. PCA test by using sera from BSA group of BN rats after both irritations showed the strong positive result characterized as large amount of subcutaneous effusions of Evans blue in SD rats, however, the sera from BSA group of GP were negative in PCA test. Serum IgE concentration did not increase after each irritation in BSA group of both BN rats and GP.

CONCLUSIONBN rats were more sensitive than GPs on initiative systemic anaphylaxis test and passive cutaneous anaphylaxis test. Meanwhile, BN rats has an advantage in experimental treatment compared with Guinea pigs.

Allergens ; administration & dosage ; toxicity ; Anaphylaxis ; chemically induced ; Animals ; Guinea Pigs ; Hypersensitivity ; etiology ; Male ; Models, Animal ; Rats ; Rats, Inbred BN ; Rats, Sprague-Dawley

The industry of germ-free animals has been a hot spot in research along with the rapid development of studies on the relationship between microbiota and host diseases. Because it is pathogen?free, and the high degree of simi?larity in anatomy, physiology, pathogenesis to humans, germ?free pig is considered a clinical relevant model to be widely used in life science research. Based on the current state of research of germ?free pig cultivation at home and abroad and the experimental studies carried out in our laboratory as well, this article gives a simple discussion on germ?free technique of domestic pigs.

OBJECTIVETo investigate the mercury cumulation following single dose or long-term use of Cinnabar to rats.

METHODThe Cinnabar which was used in the study contains 98% insoluble mercuric sulfide (HgS) and 21.5 mg x kg(-1) soluble mercuric compounds. Two separate experiments were performed: (1) Tweenty-eight fasting SD rats were orally given a single dose of Cinnabar at the dose of 0.8 g x kg(-1) and the other four rats were given ultra-filtrated water served as control group. Blood, livers, kidneys and brains of four rats were taken out at 0.5, 1, 2, 4, 8, 16, 36 h respectively after treatment. Mercury quantity of each organ or blood sample was measured. (2) Forty SD rats were randomly divided into four groups: control group and Cinnabar 0.1, 0.4, 0.8 g x kg(-1) groups, each group containing 5 females and 5 males. The rats were intra-gastrically treated with Cinnabar once a day for successively 90 days, while the control group was given ultra-filtrated water. Mercury contents in blood, livers, kidneys and brain of each rat were measured at 16 h of fasting after last dosing.

RESULTMercury contents of blood, liver, kidney and brain increased slightly after single dosing of Cinnabar at dose of 0.8 g x kg(-1), with the order from high to low liver > blood > brain > kidney. Whereas 90-day oral treatment of Cinnabar led to significant cumulation of mercury in organs but not in blood. Kidney' s cumulation of mercury was much higher than any other tested organs and blood. Brain's mercury cumulation was also very high. The contents of mercury in kidney and brain of 0.8 g x kg(-1) group (total intake of soluble mercury within 90 days was 1 548 microg x kg(-1)) were respectively 71.2 and 27.4 times higher than control group. Even though in the lowest dose 0.1 g x kg(-1) group (total intake of soluble mercury 194 microg? kg(-1)), the mercury cumulation folds in kidney and brain were 16.77 and 20.43 respectively. However, liver got lower mercury cumulation than kidney and brain, which led to only 2 folds mercury cumulation at dose of 0.8 g x kg(-1). Our previous study showed that 90-day administration of Cinnabar at the dose > or = 0.1 g x kg(-1) (total intake of soluble mercury 194 microg x kg(-1)) could cause pathological changes in kidney and liver, indicating both were the toxicity targets for Cinnabar. Those manifested that liver could be more sensitive than kidney to mercury. Though brain got 20 times mercury cumulation after 90 day treatment, the animals showed no abnormal signs in general behavior and brain histomorphology,which indicated that rat brain was not sensitive to mercury.

CONCLUSIONSoluble mercury in Cinnabar can be absorbed causing high cumulated in some organs, such as kidney and brain after long-term use of Cinnabar. Liver had also mercury cumulation, but was much lower than kidney. Total intake of soluble mercury for > or = 194 microg x kg(-1) within 90 days could cause toxicosis by mercury cumulation.

Administration, Oral ; Animals ; Brain ; metabolism ; Female ; Kidney ; metabolism ; Liver ; metabolism ; Male ; Mercury ; pharmacokinetics ; Mercury Compounds ; administration & dosage ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution

OBJECTIVETo investigate the substance basis and the mechanism of pseudoanaphylactoid reactions (PR) induced by Shuanghuanglian injection (SHLI).

METHOD(1)The study of PR and the substance basis of PR of SHLI: ICR mice were divided into different test groups, the mice were intravenously injected with solutions of different concentration of SHLI, baicalin, forsythin, caffeotannic acid, positive control Compound 48/80 and normal sodium. All test substances were mixed with 0.4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after SHLI injection. (2) The study of mechanisms: Mice were pretreated with an oral administration of Astemizol, intraperitoneal injection of cyclophosphamide 75 mg x kg(-1) or Compound 48/80 4 mg x kg(-1), then mice were intravenously injected with SHLI. At last, vascular permeability of the ears in pretreated groups was compared with SHLI treatment alone group.

RESULTSHLI of 300 mg x kg(-1) and 600 mg x kg(-1) caused obvious vascular hyperpermeability, but baicalin, forsythin and caffeotannic didn't cause vascular hyperpermeability in the ears. The Astemizol can decrease the degree of SHLI-induced vascular hyperpermeability of the ears in the mice. After intraperitoneal injected with cyclophosphamide, there was a slight decrease in the degree of SHLI-induced vascular hyperpermeability, but there was no marked changes in the degree of the SHLI-induced vascular hyperpermeability after the mice were pretreated with Compound 48/80.

CONCLUSIONSHLI in clinic equivalent dose can cause vascular hyperpermeability. Baicalin, forsythin and caffeotannic may not result in the PR of SHLI. The mechanism of the PR maybe relate to that SHLI stimulates histamine release, the activation of leucocyte maybe take part in the SHLI-induced PR, too. Antihistamine drug can prevent the genesis of PR which induced by SHLI.

Anaphylaxis ; chemically induced ; pathology ; physiopathology ; Animals ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; chemistry ; Injections ; Mice

OBJECTIVETo investigate the characteristics, sensitizin and the mechanism of pseudo allergic reaction induced by Yuxingcao injection.

METHODBeagle dogs were randomly assigned to control group, 0.5% tween 80 group, Yuxingcao injection without tween 80 group, Yuxingcao injection included 0.5% tween 80 group. The animals in control group were intravenously injected with saline. The other group were intravenously injected with the corresponding test substances. Observe pseudo anaphylaxis of Beagle dogs within 30 min after administration. Blood pressure and respiration rate of Beagle dogs were measured before and after injection drugs 10 min and 30 min respectively. The pseudo allergic reactions were scored at same time points, and the sera of animals were collected to determine the HIS, CH50 and C5b-9 concentration using ELISA.

RESULTThe scores of allergic reaction in 0.5% tween 80 group and Yuxingcao injection included 0.5% tween 80 group was evidently higher than that in control group in 2-5 min after administration. Animals of above two groups showed the symptoms of red swelling on ear part, pruritus, throwing the head, nausea, lapping the tongue, dysphoria and bradykinesia. Some of them had behaved with repose, urination, defecation, cyanosis, the frequency of breathes accelerating and blood pressure decreasing. The rate of pseudo allergic reactions was 100%. Serum CH50 concentration of 0.5% tween 80 group decreased 10 min after injection, while C5b-9 concentration increased. No obvious differences were observed 30 min after injection. There was no significant difference in HIS concentration between control group and treatment groups.

CONCLUSIONThe pseudo allergic reactions appeared after intravenous 0.5% tween 80 and Yuxingcao injection when mixed with tween 80. Furthermore, Yuxingcao injection without tween 80 did not induce pseudo allergic reactions. It was suggested that the pseudo allergic reactions of Yu Xing Cao Injection was related to the cosolvent tween 80. The pseudo allergic reactions of tween 80 may relate to the activation of complement.

Animals ; Disease Models, Animal ; Dogs ; Drug Hypersensitivity ; etiology ; immunology ; Drug Interactions ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Humans ; Male ; Polysorbates ; administration & dosage ; adverse effects ; Random Allocation

OBJECTIVETo establish a model of gastric precancerous lesion by using Aristolochic manshuriensis which contains aristolochic acids.

METHODThe SD rats were randomly divided into four groups: control and three different doses of ethanol extractive of A. manshuriensis (EEA) (corresponding to aristolochic acid I 2.5, 5.0, 10.0 mg x kg(-1)), respectively. EEA was intragastrically given to rats every other day. At the end of the 10th, 15th, 20th week, part of the rats in each group was sacrificed and the stomachs were weighed. The gastric tumor was assessed by the weight and the relative stomach weight to the body weight. The stomachs were fixed in 4% neutral formalin, and the paraffin imbedding tissues were sliced and HE stained. Histomorphology was observed under the light microscope to determine gastric hyperplasia, mucosa precancerosis (atypical hyperplasia) and gastric cancer formation.

RESULTThe rats treated with different doses of EEA for 10 weeks induced mucosa papillary, epithelioma hyperplasia. Histological observation showed mucosa precancerosis lesions characterized as atypical hyperplasia at the dose levels corresponding to aristolochic acid I 5.0 and 10.0 mg x kg(-1) treated for 10 weeks. The incidence rate of gastric precancerosis in those two groups was 100% at the 15th week. Malignant tumors were observed in most of the animals in 10.0 mg x kg(-1) group. The animals in 5.0 mg x kg(-1) group were well tolerant compared to 10.0 mg x kg(-1) group during the course of experiment, so the dose of aristolochic acid I 5.0 mg x kg(-1) and 10-15 weeks treatment were considered to be optimum to establish the model of gastric precancerosis.

CONCLUSIONA rat model of gastric precancerosis can be induced within a short duration by giving an oral administration of the ethanol extract of A. manshuriensis which contains aristolochic acids.

Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; administration & dosage ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Stomach Neoplasms ; drug therapy ; pathology

OBJECTIVETo search new effective compounds, the different hemostatic effects of Flos Sophorae, Flos Sophorae Carbonisatus and principal constituent were observed.

METHODUsing the bleeding time (BT) and the recalcification time (RT) as the specificity indicators for the hematischesis function, the hemostatic effects of the following were observed. Flos Sophorae, Flos Sophorae Carbonisatus, characteristic value extraction thing A and B (SCE A and B) and the principal constituent after orally administered in normal rats in order to analyze the new hemostatic compounds.

RESULTFlos Sophorae, and Flos Sophorae Carbonisatus can obviously reduce BT and RT in rats, in which the effect of Carbonisatus is stronger than the crude. Otherwhile, SCE A and SCE B can also obviously reduce BT and RT in rats, in which the effects of SCE B surpassed those of SCE A. Furthermore, two characteristic compounds extracted from SCE B (kaikasaponin I called compound 1 and isorhamnetin-3-O-rutinoside named called 2) and other nominated principal constituents (rutin, tannin), can obviously shorten BT and RT in rats, among which compound 2 is most superior.

CONCLUSIONThe Flos Sophorae, Flos Sophorae Carbonisatus and their character compounds can shorten! the BT and RT in rats. The compound 2 from SCE B has the most superior effect. Study showed that compound 2 should be the new hemostatic compounds after Flos Sophorae carbonized. The results also indicated that the increase of hemostatic effect after Flos Sophorae carbonized should be related with the coordination of various kinds of ingredient.

Animals ; Bleeding Time ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flowers ; chemistry ; Hemostatics ; administration & dosage ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sophora ; chemistry

OBJECTIVETo investigate the preclinical evaluation method of pseudoanaphylactoid reactions for Chinese herbal injections.

METHODBeagle dogs were divided into control group (C), 0.5% tween 80 group (T), Yuxincao injection containing 0.5% tween 80 (YT), distilled solution from Yuxincao (Y). Various groups of Beagle dogs were given 3 mL x kg(-1) of the test articles intravenously. The anaphylactoid reactions were observed immediately, while blood pressure, respiratory frequencies and heart rates were tested at 10 min and 30 min after administration.

RESULTA variety of symptoms that range from cutaneous and mucosa signs to bronchospasma and cardiovascular collapse, including angioedema at lip, conjunctiva, ear and circumoral skin, somnolence, lethargy, breathless or dyspnea, severe hypotension etc were observed in T and TY groups from immediately post-injection to at least 30 min after administration. These reactions occurred at both first injection or repeated injections at 24 weeks intervals, manifesting that it was pseudoanaphylactoid reaction mediated by non-immune mechanisms.

CONCLUSIONBeagle dogs could be used as an animal model for preclinical evaluation of pseudoanaphylactoid reactions of Chinese herbal injection with sensitivity, reproducibility, and high clinic consistency.

Anaphylaxis ; chemically induced ; etiology ; immunology ; Animals ; Dogs ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Injections, Intravenous ; Male ; Models, Animal ; Random Allocation