Objective To observe the distribution of bone marrow mesenchymal stem cells(MSCs) and the effects on expression of apoptosis relative proteins Caspase 3 and Bcl-2 after intravenous transplanted into ischemic rat brains.Methods MSCs from SD rats were cultivated and proliferated in vitro and marked with CFSE.MSCs were then intravenously transplanted into middle cerebral artery occlusion(MCAO)models of SD rats.The rats were killed at different time points to observe the distribution of MSCs under fluorescence microscoDe as well as the effects on expression of apoptosis relative proteins Caspase 3 and Bcl-2 using immunohistochemical method.Results Density of Caspase 3 in immunohistochemically positive area in transplantion group were(2.81±0.35)%,(3.98±0.67)%,(5.58±0.92)%,(3.51±0.63)%,(1.64±0.29)%in 6,12,24,72 hours and in 7 days,respectively,and decreased significantly compared with those of control group[(3.92±0.44)%,(5.23±0.30)%,(6.89±0.57)%,(4.39±0.57)%,(2.29±0.21)%],the difference being significant(t=4.37,3.34,2.60,2.32,3.90,P＜0.05 or＜0.01).The density of Bcl-2 in immunohistochemically positive area in transplantation group were(4.70±0.16)%,(5.61±0.26)%,(3.00±0.28)%respectively in 6,12 hours and in 7 days,which had improved significantly compared with those of control group[(3.28±0.27)%,(4.54±0.59)%,(2.15±0.62)%],the difference being significant(t=8.32,3.25,2.54,P＜0.05 or＜0.01).Conclusions Bone marrow MSCs can exert protective effects on brain ischemia and reperfusion injury possibly by down-regulating Caspase 3 and up-regulating Bcl-2.

China ; Heart Transplantation ; methods ; Humans

UNLABELLEDTo investigate the effects of Shizhang Cataplasm (SC) and Xuzhang Cataplasm (XC) in treating liver cirrhosis caused ascites of excessive syndrome (ES) type and deficient syndrome (DS) type respectively.

METHODSAll the 77 patients (37 of ES type and 40 of DS type) enrolled were treated by conventional treatment but with restrictive use of diuretics. SC and XC were given respectively to 26 patients of ES type and 26 of DS type additionally by umbilical sticking, they were regarded as the treated group, and those (11 of ES type and 14 of DS type) not received the cataplasm treatment were regarded as the control group. The changes of symptoms, body weight, abdominal perimeter and amount of urine before and after treatment were observed, and amount of ascites was examined with B-ultrasound to evaluate the efficacy according to comprehensive grading criteria. Also, the toxicity was observed.

RESULTSSixty-two cases completed the full course, 15 were withdrawn. As compared with the corresponding control group, body weight, abdominal perimeter and amount of ascites decreased, while amount of urine and flatus discharging increased remarkably in the treated group (P < 0.05). The comprehensive efficacy in patients of ES type was better than that in DS type (P < 0.05). The effective rate of grade I/II was 7.1% and 9.1% for patients in the control group of DS type and ES type respectively, while it was 57.2% and 69.2% in the treated group of DS and ES type respectively. Better therapeutic effect was shown in patients of ES type treated with SC.

CONCLUSIONSC and XC showed good assistant effects in treating patients with liver cirrhosis caused ascites of ES and DS type respectively.

Administration, Cutaneous ; Ascites ; diagnostic imaging ; drug therapy ; etiology ; Diagnosis, Differential ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Liver Cirrhosis ; complications ; diagnostic imaging ; drug therapy ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Ultrasonography ; Umbilicus

OBJECTIVETo investigate the release characteristics and mechanism of oridnonin self-microemulsifying drug delivery system (SMEDDS) in vitro.

METHODThe concentration of oridonin was determined by HPLC. In vitro release studies were conducted by reverse dialysis technique. The effects of release medium, agitation rate and preparations on the oridonin release were studied. The similarity factor (f2) was applied to the release profile comparisons. Model fitting was used to determine the kinetics and mechanism.

RESULTThe release media and agitation rate from 50-100 r x min(-1) had no distinctive effect on the oridonin release kinetics, which the similarity factors were greater than 50. The oridonin release profiles for oridonin SMEDDS and oridonin ethanol solution were similar. 65% of oridonin were released in 30 min for oridonin SMEDDS in pH 7.8 PBS. Oridonin SMEDDS fit the Hixson-Crowell model best.

CONCLUSIONThe release data from oridonin SMEDDS showed it release fast. The deduced release mechanism is that the surface and particle sizes of self-microemulsion in water solution are changing during the process of release and the drug penetration through membrane is a passive diffusion process.

Chromatography, High Pressure Liquid ; Diterpenes, Kaurane ; chemistry ; Drug Delivery Systems ; methods ; Emulsions ; chemistry ; Kinetics

OBJECTIVETo establish a chiro chromatography for studying the stereoselective metabolism of propranolol (PL) in S(9) incubates prepared from transgenic cell lines expressing human cytochrome P450.

METHODSThe concentration of each enantiomer in S(9) incubates was determined through precolumn derivatization with GITC, followed by RP-HPLC assay using S-(+)-propafenone as internal standard.

RESULTSBaseline separations among the diastereomers of S(-)-P, internal standard and R(+)-PL were achieved on Shimpack CLC C(18)ODS column, with UV detection and methanol:water:glacial acetic acid (67/33/0.05,v/v/v) as mobile phase. The assay was simple, accurate, precise and specific. The linear range was from 5 to 500 micromol/L for each enantiomer. The limit of quantitation (LOQ) for the method was 5 micromol/L for the S(-)-and R(+)-PL, respectively (n=5, RSD<10%). The analytical method afforded average recoveries of 98.7 and 98.1% for S(-)- and R(+)-PL, respectively. The reproducibility of the assay was good (RSD<10%). The time-dependent studies showed that PL had the stereoselectivity of S-(-)-isomer in metabolism via CYP2C18 and the stereoselectivity of R-(+)-isomer in metabolism via CYP2C9.

CONCLUSIONThe method allows to study of stereoselective metabolism of PL in vitro.

Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme System ; genetics ; physiology ; Humans ; Propranolol ; analysis ; metabolism ; Reproducibility of Results ; Stereoisomerism ; Transgenes

Adult ; Female ; Humans ; Male ; Medial Collateral Ligament, Knee ; injuries ; surgery ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Rupture ; Suture Anchors

OBJECTIVETo develop an HPLC method to determine vitexin-rhamnoside in plasma of Beagle dogs and study the pharmacokinetics and bioavailability of Yixintong sustained release tablets in Beagle dogs.

METHODA newly-developed HPLC method using C18 column and methanol-acetonitrile-tetrahydrogenfuran-0.5% acetic acid (1:1:19.4:78.6) as mobile phase was validated, and then was employed to determine vitexin-rhamnoside in plasma of Beagle dogs after oral administration of Yixintong sustained release tablets and general tablets. The main pharmacokinetic parameters were estimated by pharmacokinetic program 3p87. The non-compartmental pharmacokinetic parameters were also calculated on basis of the statistic moment theory.

RESULTThe pharmacokinetic profiles of Yixintong sustained release tablets and the general tablets were fitted to a one-and two-compartment open model, respectively. The T1/2, Tmax, AUC0-infinity and MRT for Yixintong sustained release tablets were 5.22 h, 4.0 h, 6,792.75 ng x h x mL(-1) and 8.4 h, respectively, compared with 8.94 h, 1.0 h, 5,880.4 ng x h x mL(-1) and 6.1 h for the general tablets. The relative bioavailability of the Yixintong sustained release tablets was 115.5% in Beagle dogs.

CONCLUSIONThe sustained-release characteristic of Yixintong sustained release tablets were confirmed by pharmacokinetic study.

Administration, Oral ; Animals ; Apigenin ; chemistry ; Biological Availability ; Dogs ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Male ; Plasma ; chemistry ; drug effects ; Tablets ; administration & dosage ; pharmacokinetics

Objective To investigate the characteristics of infecting pathogens,their changes and antimicrobial susceptibilities on CAPD related peritonitis in our peritoneal dialysis(PD) center in the past 15 years.Methods Two hundred and six CAPD related peritonitis episodes in 145 patients from 2000 to 2005 were analyzed and compared with 109 episodes from 1991 to 2000.The causative pathogens,their antimicrobial susceptibilities and outcomes on CAPD related peritonitis from the two periods were retrospectively reviewed and compared.Results Culture negative rate decreased from 60.6% in 1990 s to 47.6% in the last five years (P=0.031 ).Among culture positive peritonitis episodes,the incidence of gram positive bacteria (GPB) peritonitis increased from 25.6% to 39.8% (P=0.059).This was mainly due to a significant increase in coagulase-neagative staphylococcus peritonitis,which significantly increased from 4.7% to 26.9% (P=0.01).Gram negative bacteria (GNB) peritonitis decreased slightly (44.2% vs 34.3%,P=0.322).The incidence of Klebsiella pneumoniae peritonitis significantly decreased (14.0% vs 3.7%,P=0.023),while Pseudomonas aeruginosa and Escherichis coli peritonitis rates slightly increased (4.7% vs 9.3%,P = 0.338;7% vs 18.7%,P=0.072).The decrease of fungal peritonitis rate was not significant (30.2% vs 17.6%,P= 0.123).The comparison of clinical outcomes showed an improvement of total recovery rate from 68.8% in 1990 s to 73.9% for 2000-2005 (P=0.09).The catheter removal rate decreased from 19.2% to 14.3% (P=0.238),and the mortality from 10.1% to 5.4% (P=0.118).In both periods,fungal peritonitis had the poorest results,which all the patients either withdrew from PD or died.Conclusions Compared with that in 1990 s,the culture positive rate for CAPD related peritonitis in 2000-2005 has been greatly improved.Coagulase-negative staphylococcus is the most common causative pathogen.The mortality and catheter removal rate have been markedly reduced in the last five years.Fungal peritonitis is the most important reason for patients' dropout.

OBJECTIVETo observe the effects of pravastatin on platelet-derived nitric oxide system in hypercholesterolemia (HC) and atherosclerosis (AS) in rabbits, and the relationship between these changes and atherosclerosis courses.

METHODSThirty male New Zealand white rabbits were randomly divided into three groups, 12 in group A, 12 in group B, and 6 in group C. All of them were fed daily with cholesterol-rich food during the first 12 weeks. In addition, in group A, pravastatin (10 mg) was orally administered daily. At the end of the 12th week, 6 in group A and B were killed randomly and their aortas were removed and the pathologic changes were observed. In the following 12 weeks, food enriched with cholesterol was substituted with normal food in all three groups. Pravastatin treatment was continued or started in the remaining members of group A and group B, but not in group C. At the end 24th week, all rabbits were killed and their aortas were examined for the fatty-streaks or atherosclerotic plaques. The expressions of endothelial NOS (eNOS) mRNA and inducible NOS (iNOS ) mRNA, NOS activity, NO production and the level of the serum lipids were measured at 0, 6th, 12th, 18th and 24th week.

RESULTSThe expression levels of platelet-derived NOS mRNA, eNOS mRNA ratio in group A had no difference at above time points, while in group B were reduced significantly at 6th week and 12th week compared with at 0 week (P <0.01), and increased at 18th week and 24th week compared with 12th week (P <0.05). The expression levels of eNOS mRNA in group C were reduced at 6th, 12th and 18th, 24th week compared with 0 week (P <0.05 and P <0.01, respectively), and were reduced in groups B and C compared with group A at 6th ,12th week (P < 0.05) and increased in group A and B compared with group C at 18th, 24th week (P <0.01). The expression levels of iNOS/mRNA among the three groups had no difference. Pathologic finding of the arteries: AS was not found in group A from the 12th to 24th week. While in group B, there were a lot of fatty-streaks on the entire intima of all large arteries at the 12th week. There were also fatty-streaks in the ascending aorta, but were improved at the 24th week. In group C, there were marked plaques in the entire aorta at the 24th week.

CONCLUSIONSThe expressions of platelet-derived eNOS mRNA, NOS activity, NO production are decreased in HC or AS rabbits. Pravastatin can up-regulate expressions of platelet-derived eNOS mRNA, NOS activity, leading to preventing or improving the pathological courses of AS.

Animals ; Atherosclerosis ; blood ; pathology ; Blood Platelets ; metabolism ; Disease Models, Animal ; Male ; Nitric Oxide ; blood ; genetics ; Nitric Oxide Synthase ; blood ; genetics ; Pravastatin ; pharmacology ; RNA, Messenger ; genetics ; Rabbits

This study was aimed to clarify whether valproic acid (VPA) induces apoptosis of leukemia HL-60 cell line and its possible mechanism. The effect of different concentrations and treatment time of VPA on HL-60 cell proliferation was assayed by cytotoxicity test (CCK-8 method) and fluorescence microscopy, and flow cytometry was used to detect cell apoptosis. The expressions of telomerase subunit h-tert mRNA and apoptosis-related protein as well as caspase-3 activity were detected by real time-quantitative PCR, Western blot and ELISA respectively. The results indicated that VPA inhibited proliferation of HL-60 cells and induced cell apoptosis in a dose dependent manner (r = -0.87). The expressions of anti-apoptotic protein BCL-2 and h-tert mRNA were significantly decreased while the pro-apoptotic protein BAX and caspase-3 activity increased after treatment with VPA. The apoptosis rate of HL-60 cell was negatively correlated with expression of h-tert mRNA. It is concluded that VPA can inhibit leukemia HL-60 cell proliferation and induce apoptosis. The VPA displays anti-leukemia activity possibly through reducing h-tert mRNA and BCL-2 protein expression, increasing BAX expression and activity of caspase-3.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Telomerase ; metabolism ; Valproic Acid ; pharmacology ; bcl-2-Associated X Protein ; metabolism