Objective: To optimize the water extracting and refining procession of Sijunzi Decoction Granules (SDG). Methods: The orthogonal test method was used to study the four factors including the soak time, amount of water added, extraction time and frequency by taking the dry extract yield of the medicinal materials and contents of ginsenoside Rg1, Re, Rb1, glycyrrhizic acid, and liquiritin as indicators to optimize the water extracting process of SDG. To optimize the alcohol precipitation process of SDG, the factors including the concentration of the medicinal materials, alcohol content, and time of alcohol precipitation were investigated. The extracts before and after alcohol precipitation were compared by intervening spleen deficiency syndrome pharmacodynamics experiment. Results: The best water extracting procession of SDG was soaked for 60 min with 10 times of water, decocted three times, 30 min for each time. The optimal water extraction by alcohol sedimentation process was to concentrate the filtrate of water extraction to 1 mL, which was equivalent to 0.5 g of the original medicinal materials, with 80% alcohol content and 12 h alcohol precipitation time. Compared with the model group, the activity of salivary amylase and the conent of serum gastrin in the water extracting group and water extracting by alcohol sedimentation group of SDG were significantly increased and could improve the absorption function of xylose in small intestine of rats with spleen deficiency syndrome (P < 0.05); Compared with the water extracting group of SDG, there was a significant difference in D-xylose excretion rate in the water extracting by alcohol sedimentation group of SDG (P < 0.05), but there was no significant difference in activity of salivary amylase and content of serum gastrin (P > 0.05). Conclusion: The alcohol precipitation process is not suitable for refining of SDG. So the water extracting process is finally selected as the best extraction process of SDG.

BackgroundHigh myopia is one of leading causes of blindness,so far the pathogenesis remains unclear.Two single-nucleotide polymorphisms (SNPs) of rs6885224 and rs12716080 in CTNND2 gene were recently found to be associated with high myopia in Singaporean Chinese.But whether these SNPs are related with the pathogenesis of high myopia in Han Chinese is worth studying,Objective This study was to investigate the relationship between the genetic variations of the CTNND2 gene and high myopia in Han Chinese.MethodsA case-controlled association study was designed.Nine hundred and thirty-three individuals with high myopia and 1227age- and gender-matched normal subjects were included in this study.The 5 ml of periphery blood was obtained from all subjects for the extraction of genomic DNA.The target DNA was amplified using PCR and purified by the SNaPshot method.Four SNPs rs12716080,rs917012,rs6885224 and rs16901340 in the CTNND2 gene were genotyped.This study was approved by the Ethic Committee of Sichuan Provincial People Hospital.Written informed consent was obtained from each subject before his/her enrollment.Results The frequencies of the genotypes rs6885224,rs12716080,rs917012,rs16901340 SNPs were in Hardy-Weinberg equilibrium (HWE) ( P=0.181,0.085,0.732,0.313,0.264,0.663,0.084,0.196).There were no significant differences in genotypes frequency distribution ( in turn P =0.654,0.406,0.828,0.403 ) and allele frequency distribution of the CTNND2 gene ( in turn P =0.377,0.209,0.743,0.198) between the high myopia group and normal control group.The haplotypes (TA and GA)frequencies of rs12716080 and rs917012 in the high myopia group were significantly different from those of the normal control group(TA:0.784 vs.0.719;GA:0.087 vs.0.136) (x2 =6.115,P=0.013 ;x2 =6.634,P=0.010),but those of GG were similar between the high myopia group and normal control group ( 0.123 vs.0.143,x2 =0.889,P =0.346). ConclusionsThe SNPs rs12716080,rs917012,rs6885224 and rs16901340 in CTNND2 gene were not responsible for high myopia,however,the haplotypes of rs12716080 and rs917012 are susceptible for high myopia in Han Chinese.

To evaluate the value of left ventricular diastolic function in type 2 diabetes mellitus (DM) using dual-gate Doppler and relative factors, we included 50 non-obesity and hypertension-free DM patients into the controlled group in the study along with 48 age-and-gender-matched healthy volunteer subjects. The peak early diastolic velocity (E), peak later diastolic velocity (A), deceleration time (DT), anterior mitral annulus diastolic peak velocity (e') , isovolumic relaxation time (IVRT), E/A, E/e', Tei index and T(E-e), were measured with dual-gate Doppler. 20 subjects were randomly selected for repetitive analysis. Study showed statistical difference in E/A, DT, e', E/e', IVRT, Tei index and T(E-e), between the two groups (P < 0.05). Linear regression analysis showed positive correlation between T(E-e), and IVRT, course of DM patients and T(E-e), (Beta = 0.295, P = 0.020), and HbA1c control level and T(E-e), (Beta = 0.399, P = 0.010). Repeated analysis showed good reproducibility for both within and between groups. Dual-gate Doppler has clinical value in evaluating left ventricular diastolic function in type 2 diabetes mellitus patients. The course of type 2 diabetes mellitus patients and HbA(1C) control level were both closely related with left ventricular diastolic function.
Case-Control Studies ; Diabetes Mellitus, Type 2 ; physiopathology ; Diastole ; Echocardiography, Doppler ; Humans ; Hypertension ; Mitral Valve ; Regression Analysis ; Reproducibility of Results ; Ventricular Dysfunction, Left ; Ventricular Function, Left

AlM:To explore the effective method for the treatment of complicated ocular trauma accompanied with cyclodialysis.METHODS: Sixty - eight cases of complexity ocular trauma with cyclodialysis in different degrees were undergone vitrectomy and ( or ) combined with ciliary body reduction.RESULTS:All patients were followed up for 10 ~36mo (mean 17. 0±5. 7mo). The postoperative visual acuity was better than that of preoperation (P<0. 05). Compared with preoperative, intraocular pressure was significantly increased ( P < 0. 05 ). Successful rate of ciliary body restoration was 91%. CONCLUSlON: For the complicated ocular trauma accompaniedwith cyclodialysis, vitrectomy and ( or ) ciliary body reduction is an effective treatment method.

To investigate the effect of Jiawei Foshou San and its various combined administration on hepatic P450 enzyme activity and hepatocyte morphology in rats. Rats were orally administered with drugs for four weeks and then sacrificed to prepare liver microsomes. The liver microsomes were incubated with the cocktail method; The metabolites were determined with the rapid liquid chromatography with tandem mass spectrometry (LC-MS/MS) to investigate the hepatocyte P450 enzyme activity. In addition, the hepatic pathological changes were observed by using the hematoxylin and eosin (HE) staining. Compared with the control group, the enzyme activity of CYP1A2 and CYP3A4 in the Jiawei Foshou san group showed a significant rise (P < 0.05); the enzyme activity of CYP1A2, CYP2C9, CYP2D6, CYP2E1 and CYP3A4 in the ferulic acid + ligustrazine group and the ligustrazine + tetrahydropalmatine group showed a significant rise (P < 0.05) ; the enzyme activity of CYP1A2, CYP2D6 and CYP2E1 in the ligustrazine group showed a significant rise (P < 0.05); the enzyme activity of CYP3A4 in the ferulic acid group showed a significant reduction (P < 0.05). After the administration with various drugs, the hepatocyte morphologies in the ferulic acid group and the ligustrazine group were normal. The pathological changes were observed in the tetrahydropalmatine group, such as unclear boundary of hepatic lobules, disordered hepatic cell arrangement, blurred edge, anisokaryosis and infiltration of inflammatory cells. The ferulic acid + tetrahydropalmatine group, the ligustrazine + tetrahydropalmatine group and the Jiawei Foshou San group also showed inflammatory infiltration, but with less pathological changes, particularly the Jiawei Foshou San group. The study result shows that Jiawei Foshou San can induce the enzyme activity of CYP1A2 and CYP3A4, and ligustrazine may be the effective substance for inducing CYP1A2. Its combination with ferulic acid and ligustrazine can significantly reduce the liver toxicity of tetrahydropalmatine.
Animals ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; enzymology ; Microsomes, Liver ; drug effects ; enzymology ; Rats ; Rats, Sprague-Dawley

To evaluate the clinical efficacy of recombinant human Interleukin 11 in the treatment of pre-aplastic anemia, six patients with pre-aplastic anemia were injected with rhIL-11 of 6 million units once a day during 7-14 days. Blood platelet counts were taken on day 8, 15, 30 and 60 after the treatment, and bone marrow examination was performed on day 15 as compared with those before treatment. The results showed that platelet counts in 3 out of 6 patients increased remarkably (50%), one of the six increased moderately (16.7%), another case of the six increased slightly (16.7%), platelet in one out of six did not significantly increase (16.7%), the total efficacy rate is 83.3%, the amount of megakaryocyte in bone marrow of all six patients increased, the side effect of the rhIL-11 treatment was light. In conclusion, the efficacy of recombinant human Interleukin-11 in the treatment of thrombocytopenia patients with pre-aplastic anemia is satisfactory. As the number of the cases is too small to conclude, further exploration needs accumulation of more applications.
Adult ; Aged ; Anemia, Aplastic ; drug therapy ; pathology ; Chronic Disease ; Female ; Humans ; Interleukin-11 ; genetics ; therapeutic use ; Male ; Middle Aged ; Platelet Count ; Recombinant Proteins ; therapeutic use ; Thrombocytopenia ; blood ; drug therapy ; Treatment Outcome

OBJECTIVETo study the mechanism of uPA improving sperm capacitation by investigating the effect of uPA on the mitochondrial function of mouse capacitated-sperm in vitro.

METHODSMitochondrial function of mouse capacitated-spermatozoa was evaluated through the assessment of mitochondrial membrane potential using JC-1 performed by flow cytometer and fluorescent microscope respectively. The experiment and the control groups were designed according to the presence or absence of uPA, each divided into 5 subgroups based on the different time of uPA treatment (or BWW in the control groups) at 0, 5, 15, 30 and 60 min respectively.

RESULTS(1) Compared with that at 0 min, the mean fluorescence intensity of JC-1 within the spermatozoal body and the percentage of orange-red colored spermatozoa in the experiment group were increased significantly at 5 and 15 min respectively after uPA incubation (P < 0.05). (2) The mean fluorescence intensity of JC-1 within the spermatozoal body at 15, 30 and 60 min and the percentage of orange-red colored spermatozoa at 5 and 15 min in the group were significantly higher than those in the control group (P < 0.05).

CONCLUSIONuPA could increase the mitochondrial membrane potential of mouse capacitated-spermatozoa in vitro, and maintain it at a high level for a certain period of time. By enhancing sperm mitochondrial function, uPA may provide sufficient energy for capacitated-spermatozoa to increase their motility and change their motor pattern, which might be one of the therapeutic mechanisms of uPA on male infertility.

Animals ; Flow Cytometry ; Fluorescent Dyes ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mice, Inbred Strains ; Sperm Capacitation ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Urokinase-Type Plasminogen Activator ; pharmacology

BACKGROUNDTissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study was designed to develop a tissue engineering heart valve by using human umbilical cord blood-derived endothelial progenitor cells (EPCs) and decellularized valve scaffolds.

METHODSDecellularized valve scaffolds were prepared from fresh porcine heart valves. EPCs were isolated from fresh human umbilical cord blood by density gradient centrifugation, cultured for 3 weeks in EGM-2-MV medium, by which time the resultant cell population became endothelial in nature, as assessed by immunofluorescent staining. EPC-derived endothelial cells were seeded onto the decellularized scaffold at 3 x 10(6) cells/cm(2) and cultured under static conditions for 7 days. Proliferation of the seeded cells on the scaffolds was detected using the MTT assay. Tissue-engineered heart valves were analyzed by HE staining, immunofluorescent staining and scanning electron microscopy. The anti-thrombogenic function of the endothelium on the engineered heart valves was evaluated by platelet adhesion experiments and reverse transcription-polymerase chain reaction (RT-PCR) analysis for the expression of endothelial nitric oxide synthase (eNOS) and tissue-type plasminogen activator (t-PA).

RESULTSEPC-derived endothelial cells showed a histolytic cobblestone morphology, expressed specific markers of the endothelial cell lineage including von Willebrand factor (vWF) and CD31, bound a human endothelial cell-specific lectin, Ulex Europaeus agglutinin-1 (UEA-1), and took up Dil-labeled low density lipoprotein (Dil-Ac-LDL). After seeding on the decellularized scaffold, the cells showed excellent metabolic activity and proliferation. The cells formed confluent endothelial monolayers atop the decellularized matrix, as assessed by HE staining and immunostaining for vWF and CD31. Scanning electron microscopy demonstrated the occurrence of tight junctions between cells forming the confluent monolayer. Platelets adhesion experiments suggested that the neo-endothelium was non-thrombogenic. The expression levels of eNOS and t-PA genes in the neo-endothelium were quite similar to those in human umbilical vein endothelial cells.

CONCLUSIONSEPCs isolated from the human umbilical cord blood can differentiate into endothelial cells in vitro and form a functional endothelium atop decellularized heart valve scaffolds. Thus, EPCs may be a promising cell source for constructing tissue-engineered heart valves.

Animals ; Cell Proliferation ; Endothelial Cells ; cytology ; metabolism ; Heart Valve Prosthesis ; Heart Valves ; cytology ; metabolism ; ultrastructure ; Humans ; Immunohistochemistry ; Microscopy, Electron, Scanning ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; Platelet Aggregation ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Swine ; Tissue Engineering ; methods ; Tissue Plasminogen Activator ; genetics ; metabolism ; Umbilical Cord ; cytology

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Coal Mining ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Lung ; chemistry ; pathology ; Male ; Middle Aged ; Occupational Diseases ; metabolism ; Pneumoconiosis ; metabolism ; Tumor Suppressor Proteins ; analysis

Objective To investigate the changes of the expressions of ATP-binding cassette transporter A1(ABCA1)and the retinoid X receptor(RXR?in human carotid atherosclerotic plaques and to explore the possible mechanisms by which ABCA1 affects the formation of carotid atherosclerosis(CAS). Methods 24 carotid atherosclerotic plaque and 10 intestinal artery specimens were respectively collected to compared the expression levels of ABCA1 mRNA.RXR?mRNA and those protein,ABCA1 and RXR?gene expressions were determined by reverse transcriptase polymerase chain reaction(RT-PCR)in the specimens,meanwhile the presence of ABCAI and RXRcprotein was assessed by Western blot.Results ABCA1(0.79?0.04)and RXR?(0.73?0.04)gene expression were significantly elevated in carotid atherosclerotic plaques(P