To study the toxicokinetics of bakuchiol, hepatic and renal toxicity in rats after single oral administration of Psoraleae Fructus and combined with Glycyrrhizae Radix et Rhizoma, in order to provide scientific evidences for clinical safe medication use. A total of 35 SD rats were randomly divided into seven groups: vehicle (distilled water) control group, Glycyrrhizae Radix et Rhizoma group, positive control (aristolochic acid A) group, Psoraleae Fructus (40 g x kg(-1)) group( both male and female rats), Psoraleae Fructus and Glycyrrhizae Radix et Rhizoma (40 +20) g x kg(-1) group (both male and female rats). HPLC-UV method was used to determine the concentration of bakuchiol in rat plasma at different time points after single oral administration. Plasma alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), plasma creatinine (Cr), N-acetyl-β-D-glucosaminidase (NAG) and kidney injury molecule 1 (Kim-1) were measured after administration for 24 h. The main toxicokinetics parameters of bakuchiol in rats exert significantly gender difference. When Psoraleae Fructus combination with Glycyrrhizae Radix et Rhizoma, the total area under the plasma concentration-time curve( AUC), C(max), and plasma clearance (CL) of bakuchiol were increased, respectively; CL, half-life (t½) were decreased, and T(max) were prolonged. The biochemical indicators (including ALT, AST, BUN, Cr and KIM-1 level) in different dose of Psoraleae Fructus groups, were found no statistically significant difference when compared with vehicle control group. The level of NAG in both Psoraleae Fructus and compatibility with Glycyrrhizae Radix et Rhizoma groups were significant increased (P < 0.05). There are obvious effects on toxicokinetics of bakuchiol in rats when Psoraleae Fructus combined with Glycyrrhizae Radix et Rhizoma. Renal toxicity induced by Psoraleae Fructus at high dose was observed after single oral administration and no liver damage in rats was found.
Administration, Oral ; Animals ; Female ; Glycyrrhiza ; toxicity ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Phenols ; pharmacokinetics ; toxicity ; Psoralea ; toxicity ; Rats ; Rats, Sprague-Dawley ; Rhizome ; toxicity ; Toxicokinetics

A biosurfactant-producing strain(S_6)was isolated from oil-containing wastewater in oxidation ditch and identified as Pseudomonas aeruginosa based on physiological and biochemical experiments and 16S rDNA sequence analysis.Infrared spectrum analysis revealed that S_6 produced glucolipid in the process of metabolism.It was observed that S_6 decreased the surface tension of water from 72 mN/m to 33.9 mN/m with the critical micelle concentration(CMC)of 50mg/L.The measurement of oil displacement and surface tension demonstrated that the fermented liquid had stable surface activity at varying range of salinity,pH,amount of dissolved oxygen.The optimal culture condition was obtained through orthogonal experiment:glucose 10g/L,urea 5g/L,KH_2PO_4 1g/L,liquor of microelement 2mL,pH 8.0,water 1000mL;and the biosurfactant production under optimal culture condition was 0.173g/L.

Objective To evaluate the value of low-density lipoprotein-cholesterol (LDL-C) and non-high-density lipoprotein-cholesterol (non-HDL-C) in differential diagnosis of familial hypertriglyceridemia (FHTG) and familial combined hyperlipidemia (FCHL).Methods We recruited 9 FHTG pedigrees (94 subjects) and 24 FCHL pedigrees (94 subjects) and then divided them into affected groups and non-affected groups according to lipid abnormality.Another 10 normal control pedigrees (57 subjects) served as controls.We compared the routine lipid levels such as triglyceride (TAG),total cholesterol (TC),HDL-C and LDL-C and non-HDL-C between the groups.After stratification based on TAG level,we observed the relationship between LDL-C and non-HDL-C.Last we confirmed and analyzed the cut-off value of differential diagnosis between FHTG and FCHL with receiver operating characteristic (ROC) curve.Results The levels of TAG,TC,and non-HDL-C were significantly higher in the affected group of FHTG than in the non-affected group of FHTG and the normal group (P＜0.01 or P＜0.05).The levels of TAG,TC,HDL-C,LDL-C and non-tHDL-C wcrc significantly higher in the affected group of FCHL than in the non-affected group of FCHL and the normal group (P＜0.01 or P＜0.05).The levels of TAG were significantly higher (P＜0.01) while TC,HDL-C,LDL-C and non-HDL-C levels were significantly lower (P＜ 0.01 or P＜0.05) in the affected group of FHTG than in the affected group of FCHL.The association between LDL-C and non-HDL-C was positive both in FHTG and FCHL,but the relationship became weaker as TAG level increased.The cut-off value of LDL-C and non-HDL-C was 3.575 mmol/L and 4.525 mmol/L,respectively.Conclusion In addition to the routinely used lipid indexes,non-HDL-C may be a new index for differential diagnosis of FHTG and FCHL,and may be superior to LDL-C in this regard.

Objective To study the influence of Rhizoma typhonii extract on the growth of glioma SHG-44 cells cultured invitro,and to explore the mechanism of the inhibitory effect of Rhizoma typhonii extract on the growth of glioma cells.Methods The SHG-44 cells were cultured and divided into blank control group and 8, 40, 200, 1 000μg·L-1 Rhizoma typhonii extract groups.The inhibitory effect of Rhizoma typhonii extract on the growth of glioma SHG-44 cells was measured by MTT assay. The secretion levels of Bax and caspase-3 proteins were examined by ELISA assay. The expression level of caspase-3 protein was examined by Western blotting method. Results Compared with blank control group, the inhibitory rates of the growth of SHG-44 cells in 200, 1 000μg·L-1 Rhizoma typhonii extract groups at 24 h,and 8,40,200,and 1 000μg·L-1 Rhizoma typhonii extract groups at 48 h were significantly increased (P<0.05 or P<0.01).The secretion levels of Bax and caspase-3 proteins in 40,200,and 1 000μg· L-1 Rhizoma typhonii extract groups at 48 h were increased compared with blank control group (P<0.05 or P<0.01 ). Compared with blank control group, the expression levels of caspase-3 protein in different doses of Rhizoma typhonii extract groups were increased significantly (P<0.01). Conclusion Rhizoma Typhonii extract can inhibit the growth of cells through up-regulating the expression of Bax protein,increasing the expression level of caspase-3 protein and activating apoptosis pathway.

Atherosclerotic cardiovascular diseases(ASCVD)are the major causes of morbidity and mortality worldwide. Previous randomized controlled trials confirm that statin therapy can effectively reduce the level of low density lipoprotein cholesterol(LDL-C),all-cause and cardiovascular disease mortality in patients with and without ASCVD.However,there is no widespread use of lipid-lowering therapy to achieve the benefit in high risk patients with ASCVD and patients without ASCVD. Therefore, it is necessary to further elaborate the clinical benefits of statins and their combined use for lipid regulating therapy and increasing the beneficiaries.

AIMTo study the feasibility of long-term potentiation(LTP) recording in the CA1 area of the rat in vivo with electrodes-binding technique.

METHODSAnesthetizing Wistar rats with urethane and fixing the animal on the stereotaxic device for acute surgery; implanting cannula into lateral cerebral ventricle; inserting self-made bound stimulating/recording electrodes into hippocampal CA1 area; recording basal field excitatory postsynaptic potential (fEPSP) and tetanus-induced long term potentiation (LTP).

RESULTSfEPSPs were reliably induced by using the stimulating/recording electrodes-binding technique, and the appearance rate of fEPSP was nearly 100%; basal fEPSP recording was very stable, lasting for long time enough to finish all experiment; high frequency stimulation (HFS) successfully induced LTP, which maintained more than three hours, the inductivity is about 67%; paired-pulse facilitation (PPF) recording was also stable; intracerebroventricular (i c v) injection of amyloid beta suppressed HFSinduced LTP evidently.

CONCLUSIONThe electrodes-binding technique for recording hippocampal LTP in vivo is quite simple and convenient. The experimental resource can be saved, and the rates of fEPSP appearance and LTP induction are kept high. Therefore, it is promising for this technique to be one electrophysiological auxiliary method in the research of learning and memory.

Animals ; Electric Stimulation ; methods ; Electrodes ; Excitatory Postsynaptic Potentials ; physiology ; Feasibility Studies ; Hippocampus ; physiology ; Long-Term Potentiation ; physiology ; Male ; Rats ; Rats, Wistar

Objective To observe the effects of five flavonoids include rutin(RU),dihydromyricetin(DMY),hesperetin(HP),daidzein(DA)and hydroxysaffor yellow A(HYSA)on myocardiocyte apoptosis induced by H2O2 and to explore their relationships with Keshan disease and the possible mechanism.Methods Primary cultured cardiocytes of neonatal rats were randomly divided into control group,model group,and flavonoids preincubation group.The cardiocyte apoptosis was examined by fluorescent staining,the rates of apoptosis were detected by flow cytometry,the expression of Bcl-2 family proteins associated with apoptosis were observed:by Western blot.Results Compared with model group[(24.33±6.51)%],RU[(13.95±3.80)%],DA[(11.82±3.50)%],HYSA[(12.33±3.78)%]could decreased the rate of apoptosis(P＜0.05).The five flavonoids could up-regulate Bcl-2 expression,down-regulate Bax expression,and increase the Bcl-2/Bax ratio[RU(0.989±0.094),DMY(0.931±0.280),HP(0.980±0.095),DA(1.049±0.092),HYSA(1.031±0.039),vs model(0.490±0.046),the difference had statistical significances(P＜0.05)],but the Bcl-xl did not significantly changed(P＞0.05).Conclusions RU,DMY,HP,DA and HYSA have antiapoptotic effects on cardiomyocyte via regulating Bcl-2 and Bax,which gives us a hint in prevention and treatment of Keshan disease.

Objective To evaluate the rat model of type 2 diabetes mellitus ( T2DM) associated with depres-sion by body weight, fasting blood glucose, manifestations, and open field test.Methods The T2DM rat model was induced by high fat diet and low dose of STZ injection, and in addition, the T2DM rats were made restraint stress for 21 days.32 Wistar rats were randomly divided into three groups: normal group ( group N) , T2DM group ( group T)and T2 DM with depression group ( group T +D) , with 8 rats in each group.After the model was established, to measure the body weight, fasting blood glucose at day 0, 7, 14, and 21, and observe the gross manifestations, drink-ing and diet, feces, urine, and mental state, and test the rat depression by open field test.Results After establish-ment of the T+D rat model, the rats in group T+D showed some symptoms, including messy dark and gloomy hair, slow movement, increasing drinking, diet, feces and urine and mental fatigue.At day 0, 7, 14, and 21, compared with the group N, the body weight of the group T and group T+D were decreased, showing a significant difference ( P<0.05;P<0.01).At day 0, 7, 14, and 21, compared with the group N, and the fasting blood glucose in the groups T and T+D were increased, with a significant difference ( P<0.01 ) .After 21 days of restraint stress, the fasting blood glucose in the group T+D was significantly higher than that in the group T ( P<0.01 ) .Compared with the group N, the total movement distance in 5 minutes in the group T+D was reduced, but without a significant differ-ence (1532.6 ±126.8 cm vs.940.5 ±208.3 cm, P>0.05).Compared with the group N, the movement speed in 5 minutes in the group T was significantly slower than that in the group T , with a significant difference ( 5.1 ±0.4 cm/s vs.2.9 ±0.6 cm/s, P<0.05 ) , and even more slower than that in the group T +D, with a significant differ-ence (5.1 ±0.4 cm/s vs.2.4 ±0.5 cm/s, P<0.01) .Conclusions A rat model of type 2 diabetes mellitus as-sociated with depression has been successfully established by high fat diet and injection of low dose streptozotocin in combination with restraint stress for 21 days.This rat model is useful for further relevant studies.

Objective To establish a method to rapidly remove glycerol in frozen red blood cells(RBC).Methods The glycerol in frozen RBC was removed by using three-step method as following:the frozen RBC were diluted by adding salt solution with high concentration.The salt solution was removed through centrifugation.The RBC were resuspended in normal saline of corresponding volume.Three-step method could acbieve the quick removal of glycerol by controlling tbe concentration salt water and action time.The indexes of hemoglobin,free hemoglobin,glycerol residue,residual white blood cell(WBC) and hematocrit of frozen-deglycerolized RBC were compared between conventional and improved methods.Results The traditional method to remove glycerol needed 1h approximately,while the three step method only needed 15 mins approximately,greatly shortening the time and improving the efficiency.The related tests for removing glycerol RBC,except hemoglobin content was slightly lower than the "The quality requirements for whole blood and blood components (2012 edition)",the rest of them all meeted the requirements.Conclusion The three-step method has the advantages of short time and high cfficiency.However,it still needs to be further improved and optimized to make the quality of product meet the national standard.

OBJECTIVETo compare changes of hypothalamus-pituitary-adrenal axis (HPAA) in different rat models of Gan stagnation (GS), Pi deficiency (PD), Gan stagnation Pi deficiency (GSPD) syndromes, and to observe interventional effect of Chaishu Sijun Decoction (CSD, capable of soothing Gan-qi invigorating Pi) on them.

METHODSSeventy Wistar rats were divided into the normal control group (group 1), the GS group (group 2), the PD group (group 3), the GSPD group (group 4), the GS intervention group (group 5), the PD intervention group (group 6), and the GSPD intervention group (group 7) according to random digit table, 10 in each group. Rats in group 1 received no treatment. Rats in group 2 and 5 were modeled by chronic restraint method. Rats in group 3 and 6 were modeled by excess fatigue plus alimentary abstinence method. Rats in group 4 and 7 were modeled by chronic restraint, excess fatigue, and alimentary abstinence method. At the 2nd weekend of modeling, CSD at 2.86 g/kg was fed to rats in group 5, 6, and 7 by gastrogavage for 2 successive weeks. Equal volume of distilled water was given to rats in the rest 4 groups. On the 29th day, rats were killed, adrenal weight weighed, and adrenal index calculated. Levels of plasma and hypothalamus corticotropin-releasing hormone (CRH), plasma and pituitary adrenocorticotrophic hormone (ACTH), and plasma corticosterone (CORT) were determined using radioimmunity.

RESULTSCompared with group 1, adrenal index significantly decreased in group 2, 3, and 4 (P < 0.05). Of them, plasma and hypothalamus CRH, plasma CORT increased significantly in group 2 and 4 (P < 0.05). Besides, plasma and pituitary ACTH increased in group 4 (P < 0.05). Plasma and pituitary ACTH, as well as plasma CORT decreased significantly in group 3 (P < 0.05). Compared with group 2, 3, and 4, adrenal index increased significantly in group 5, 6, and 7 (P < 0.05). Compared with group 2, plasma CORT, hypothalamus CRH, and pituitary ACTH decreased significantly in group 5 (P < 0.05). Compared with group 3, plasma ACTH and CORT increased significantly in group 6 (P < 0.05). Compared with group 4, plasma CRH, ACTH, CORT, hypothalamus CRH, and pituitary ACTH decreased in group 7 (P < 0.05).

CONCLUSIONSThe function of HPA .axis was damaged to varying degrees in rats of the three models in this experiment. Hyperactivity of HPA axis existed in GS syndrome and GSPD syndrome. Impairment of feedback regulation in hypothalamus and pituitary was accompanied in GSPD syndrome. Hypofunction of HPA axis existed in PDS. CSD, capable of soothing Gan-qi invigorating'Pi, showed improvement on disarranged HPAA, but with optimal effect on GSPD syndrome. CSD had higher correlation with GSPD syndrome.

Adrenocorticotropic Hormone ; metabolism ; Animals ; Corticosterone ; Corticotropin-Releasing Hormone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypothalamo-Hypophyseal System ; metabolism ; Hypothalamus ; metabolism ; Medicine, Chinese Traditional ; Models, Animal ; Pituitary Gland ; metabolism ; Pituitary-Adrenal System ; metabolism ; Rats ; Rats, Wistar