To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.
Animals ; Benzaldehydes ; administration & dosage ; blood ; pharmacokinetics ; Benzofurans ; administration & dosage ; blood ; pharmacokinetics ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Caffeic Acids ; administration & dosage ; blood ; pharmacokinetics ; Catechols ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Diterpenes, Abietane ; administration & dosage ; blood ; pharmacokinetics ; Hydroxybenzoates ; administration & dosage ; blood ; pharmacokinetics ; Injections, Intravenous ; Lactates ; administration & dosage ; blood ; pharmacokinetics ; Phenanthrenes ; administration & dosage ; blood ; pharmacokinetics ; Plant Preparations ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Tandem Mass Spectrometry ; methods

Objective To observe the effect and adverse reactions of iron sucrose injection in treating old patients with iron deficiency anemia(IDA). Methods Thirty-two old patients with IDA were divided into injection group(17 patients)and oral group(15 patients)by random digits table. The effects of improvement on IDA and their adverse reactions were observed. Results After treatment, hemoglobin(Hb)in the injection group and oral group was(100.18±8.71)g/L and(85.80 ± 6.92)g/L(P＜0.01), and serum iron was(23.84±5.34)μmol/L and(14.63±3.29)μmol/L(P＜0.01).There were no obvious adverse reactions in injection group, whereas there were 6 cases(40%)with gastric and colon adverse reaction in oral group. Conclusions The treatment of intravenous injection of iron sucrose can be effectively used to cure old patients with IDA. It has better effect and has lower rate of adverse reaction.

Objective　Recently,it was reported the analy sis of bone free radical by using electron spin resonance (ESR),so this article aimed to determine the changes of bone free radical in fluoresis rats.Methods　To analyze the free radical level of bone in fluoresis rats fed with different dose of calcium by ESR method.Results　The free radical of bone in the group of low ca lcium level and the group low calcium+fluorides was higher than that in the cont rol group (P<0.01),but that in normal animal feeds (calcium-rich)+fluoride group was lower than that in the control group.Conclusions　Low doses of fluoride did not resulted in the increase of free radical in calcium-rich condition at short time,but at low calcium condition,fluoride could increase the level of free radical,these suggested that simple overdose of fluoride did not change the level of free radical of bone.

Objective To investigate the place reaction of support drainage tube in cholecystojejunostomy and its pathological changes.Methods Eight pigs were randomized into 2 groups.The tube-placing cholecystojeunostomy(n=2)or choledochocholedochostomy(n=2)and non-tube-placing cholecystojejunostomy(n=2)or choledochocholedochostomy(n=2)were performed respectively.All the 8 pigs were sarcrificed 3 months after the operation for pathological examination.Results Gross anatomy catheter group showed anastomotic inflammatory hyperplasia, wall hardening hyperplasia,luminal narrow contracture and stone formation.Microscopic biopsy showed bile duct mucosa in interstitial lymphocytes, plasma cells and other chronic inflammatory cell infiltration, a large number of fibrous tissue hyperplasia.However, these features were not found in pigs without placing the tube.Conclusion The tube can not prevent the stenosis.It might promote the formation of stricture since it can stimulate inflammatory hyperplasia and tissue hardening.

Animals ; Disease Progression ; Kidney Diseases ; physiopathology ; Kidney Glomerulus ; drug effects ; pathology ; Rats ; Renal Agents ; pharmacology ; Renin-Angiotensin System ; drug effects ; Sclerosis

Objective To explore the mechanism of deep brain stimulation(DBS)therapy to Parkinson's disease(PD).Methods We produced hemi-parkinsonian rat model with stereotaxically injecting 6-OHDA to right medial forebrain bundle(MFB)and stimulated ipsilateral subthalamu nucleus (STN)with platinum electrodes chronically to investigate the influence of DBS to the expression of Calbindin-28,synaptophysin and tyrosine dioxydase(TH)in Striatum by Western blot.In addition,slices of bilateral PD rats after DBS were stained to observe the expression of Calbindin-28 and synaptophysin in substantia nigra by Immunohistochemistry.Results High frequency stimulation impaired the rotational frequency 31% of unilateral PD rats triggered by apomophine;Long-term DBS increased the expression of TH in innocent striatum of unilateral PD rats 78.6%?9.5%,since the ipsilateral striatum(lesion side) was TH depleted by 6-OHDA insults;Calbindin-28 expression in ipsilateral striatum of hemi-PD rats raised up 75.4%?15.0% and long-term DBS reduced the effect by 43.0%?7.1%,meanwhile Calbindin-28 positive neurons in substantia nigra compacta in sham,PD and DBS rats were 74.5?10.2,75.7?15.6, 33.1?7.8.However,Synaptophysin expression in substantia nigra and striatum kept stable even after long- term DBS.Conclusions Consistent to the treatment to PD patients,DBS to STN alleviated the motor disorder of PD rats,the treatment might be based on regulating the expression of Calbindin-28 and TH.

Objective To explore the activated brain region of acute epilepsy in cat model induced by pentylenetetrazol(FFZ)with manganese enhanced-functional MR imaging(ME-fMRI),and evaluate the application of ME-fMRI on localization of the activated brain.Methods Forty cats were divided into 4 groups by random number table method as epileptic A and B groups as well as control A and B groups. Cats of epileptic groups were injected with PTZ(55 mg/kg)intramuscularly,and those of control groups were injected with the saline at same dose.The behavior change in the epileptic and control group A was observed and electroencephalogram(EEG)was also undertaken.Cats of epileptic and control group B were performed ME-fMRI,and the percentage of the enhanced signal intensity was then calculated.Results After injection with PTZ(55 mg/kg)intramuscularly,epileptic seizure was all evoked,and then EEG recording showed spike-wave and polyspike-wave complexes.The neocortex of cats of epileptic group B was diffusely phanero-enhanced on ME-fMRI.The percent enhancement of signal intensity in cortex of frontal lobe,parietal lobe and occipital lobe was(34.6?5.7)% and that in cortex of temporal lobe with(22.9? 6.5)%,whereas those of control group B with(14.9?4.5)% and(11.6?3.2)% respectively.And there was significant difference between the above different localization of the brain in the two groups (t=-10.43,-5.46 respectively,P

Objective:To observe the effects of electroacupuncture (EA) of different frequencies on the expression levels of substance P (SP) and vasoactive intestinal peptide (VIP) in the colon of rats with slow transit constipation (STC).Methods:One hundred healthy male Sprague-Dawley (SD) rats were randomly divided into a normal group,a model group,a low-frequency EA group,a high-frequency EA group and a variable-frequency EA group,with 20 rats in each group.The rats in the normal group were fed with normal diet.The rats in the other groups were fed with phenethylpiperidine in the diet at a dose of 8 mg/(kg·bw) per day,for 120 d,to establish the STC model.Rats in the normal group and the model group did not receive any treatment;rats in the low-frequency EA group were treated with 2 Hz continuous wave EA,rats in the high-frequency EA group were treated with 100 Hz continuous wave EA,and rats in the variable-frequency EA group were treated with 2 Hz/100 Hz sparse-dense EA.The current intensity of the EA was determined by the slight vibration of the rat limbs without painful screaming.The intervention was performed once a day,15 min/time for continuous 15 d.After treatment,the intestinal transit function and the expression levels of SP and VIP in the colon of the rats in each group were determined.Results:After treatment,the defecation duration of the first dark stool in the model group was significantly longer than that in the normal group (P＜0.05);the defecation durations of the first dark stool in the low-frequency EA group,high-frequency EA group and variable-frequency EA group were significantly shorter than the duration in the model group (all P＜0.05);compared with the low-frequency EA group,the first dark stool defecation duration of rats in the variable-frequency EA group was significantly shorter (P＜0.05);compared with the normal group,the SP and VIP expression levels in the colon of the model group were significantly decreased (both P＜0.01);the SP and VIP expression levels in the colon of the low-frequency EA group,the high-frequency EA group and the variable-frequency EA group were significantly higher than those in the model group (all P＜0.05);compared with the high-frequency EA group,the SP expression levels in the colon in the low-frequency EA group and the variable-frequency EA group were significantly increased (both P＜0.05);compared with the low-frequency EA group,the VIP expression levels in the colon in the high-frequency EA group and the variable-frequency EA group were significantly increased (both P＜0.05).Conclusion:EA improves the intestinal function of STC model rats by regulating the expression levels of SP and VIP in rat colon.The EA stimulation with 100 Hz continuous wave,2 Hz/100 Hz sparse-dense wave shows a better improvement in the colonic transit function in STC rats,followed by 2 Hz continuous wave.

Objective To study the achievements and safety of Transcatheter arterial chemoembolization (TACE) associated Portal Vein Chemo-therapy (PVC) per-drug delivery system (DDS) program in preventing the recurrence of hepatic cell cancer (HCC) and Portal Vein Tumor Thrombus (PVTT).Methods 97 cases with HCC and PVTT were treated from Januay 2009 to January 2011.Patients with tumor or tumor thrombus were resected on all the cases and randomly divided into 3 groups.TACE,PVC per-DDS TACE and PVC per-DDS were given to group A,group B,and group C,respectively.Patients in the 3 groups were followed and compared on the Disease Free Survivals (DFS) and the accumulative survival rates,at 6 months,1 year and 2 years after the operation.Results After the surgery was completed in June,the 1-year,2-year,3-year survival rates and cummulative survival rate in group C was higher than in group A or group.Significant differences did no appeare in June but did show in 1 year after the surgery (P＞0.05) as well as in both 2 and 3 years,after the surgery (P＜0.01).Conclusion Patients with HCC and PVTT,the TACE chemotherapy in association with PVC per-DDS could increase both the DFSs and accumulative survival rates,when compared to the either single TACE or PVC per-DDS,after the tumor or tumor thrombus were resected.

OBJECTIVETo establish rhesus haploidentical hematopoietic stem cell transplantation model by nonmyeloablative conditioning, and examine the effects of mesenchymal stem cells (MSC) in haploidentical transplantation.

METHODSThe recipient haploidentical rhesus monkeys were conditioned with a nonmyeloablative regimen consisted of fludarabine, cyclophosphamide, 200 cGy total body irradiation, and rabbit anti-human thymocyte globulin. Cyclosporine A, mycophenolate mofetil and anti CD25 antibody were used for graft versus host disease (GVHD) prophylaxis. Rhesus monkeys in one group were given hematopoietic stem cell (HSC) only, while in the other group HSC combined with MSC. The differences in hematopoiesis recovery, chimerism level, and GVHD between the two groups were evaluated.

RESULTSStable chimerism could be achieved in recipient monkeys. Hematopoiesis recovery was mainly related with chimerism level. MSC seemed capable of facilitating HSC engraftment, as there were more mixed chimerism and less GVHD occurrence in the HSC combined with MSC recipient group.

CONCLUSIONA rhesus haploidentical hematopoietic stem cell transplantation model is successfully established by nonmyeloablative conditioning. MSC was of great benefit to haploidentical transplantation.

Animals ; Chimerism ; Graft vs Host Disease ; prevention & control ; Haploidy ; Hematopoietic Stem Cell Transplantation ; Macaca mulatta ; genetics ; surgery ; Mesenchymal Stem Cell Transplantation ; Models, Animal ; Transplantation Conditioning