Effective constituents of essential oil from Gleditsiae Fructus Abnormalis and anti-cerebral ischemia/reperfusion injury mechanism: based on GC-MS, network pharmacology, and experimental verification / 中国中药杂志

Based on GC-MS and network pharmacology, the active constituents, potential targets, and mechanism of essential oil from Gleditsiae Fructus Abnormalis(EOGFA) against cerebral ischemia/reperfusion(I/R) injury were explored, and the effective constituents were verified by experiment. To be specific, GC-MS was used identify the constituents of the volatile oil. Secondly, the targets of the constituents and disease were predicted by network pharmacology, and the drug-constituent-target network was constructed, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the core targets. Molecular docking was performed to investigate the binding affinity between the active constituents and the targets. Finally, SD rats were used for experimental verification. The I/R injury model was established, and the neurological behavior score, infarct volume, and pathological morphology of brain tissue were measured in each group. The content of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-alpha(TNF-α) was determined by enzyme-linked immunosorbent assay(ELISA), and the protein expression of vascular endothelial growth factor(VEGF) by Western blot. A total of 22 active constituents and 17 core targets were screened out. The core targets were involved in 56 GO terms and the major KEGG pathways of TNF signaling pathway, VEGF signaling pathway, and sphingolipid signaling pathway. Molecular docking showed that the active constituents had high affinity to the targets. The results of animal experiment suggested that EOGFA can alleviate the neurological impairment, decrease the cerebral infarct volume and the content of IL-1β, IL-6 and TNF-α, and down-regulate the expression of VEGF. The experiment verified the part results of network pharmacology. This study reflects the multi-component, multi-target, and multi-pathway characteristics of EOGFA. The mechanism of its active constituents is related to TNF and VEGF pathways, which provides a new direction for in-depth research on and secondary development of Gleditsiae Fructus Abnormalis.

Regulatory mechanism of the extracellular matrix during the development of bone-tendon junction overuse injury / 中国组织工程研究

BACKGROUND: Overuse tendinopathy is a common disease with no ideal treatment because the pathogenesis of tendinopathy is still unclear. OBJECTIVE: To explore the effects of enzymes affecting collagen synthesis on the metabolism of collagen I and collagen III during load accumulation in a model of patella-patellar tendon junction overuse injury. METHODS: Thirty-two adult female New Zealand white rabbits were randomly divided into a jumping group (n=16) and a control group (n=16). In the jump group, rabbits were subjected to jump training for 2, 4, 6, and 8 weeks, 150 jumps per day, 5 days per week. No intervention was performed in the control group. Bilateral patella-patellar tendon junction samples were taken at 24 hours after 2-, 4-, 6-, and 8- week training. The time-series changes of matrix metalloproteinase 1 (MMP1), tissue inhibitor of metalloproteinase 1 (TIMP1), collagen I, and collagen III were measured by immunohistochemistry. The study was performed with an ethic approval from the Animal Ethical Committee of Beijing Sport University, China (approval No. BSU2015022). RESULTS AND CONCLUSION: Compared with the control group, the MMP1 activity was significantly lower within the first 6 weeks of training, TIMP1 activity was significantly higher at 4-8 weeks of training, and TIMP1/MMP1 was significantly higher at 4-8 weeks of training. Compared with the control group, the expression of collagen I was significantly increased in the first 6 weeks of training, but the expression of collagen III was significantly lowered after 8 weeks of training. TIMP1 activity was significantly positively correlated with collagen I expression, and MMP1 activity was significantly negatively correlated with collagen I expression. To conclude, the patella-patellar tendon junction, during jumping load accumulation, mainly responded to load stimulation by increasing collagen I expression. In the first 2 weeks of load accumulation, the expression of collagen I was mainly increased by inhibiting MMP1 activity; in the period of 4 to 6 weeks of load accumulation, the expression of collagen I was mainly increased by increasing the activity of TIMP1 and inhibiting the activity of MMP1; and after 8 weeks of load accumulation, TIMP1 could promote collagen synthesis but with inability to antagonize the degradation of collagen I and III by MMP1. Then the expression of collagen I decreased from the peak level after 6 weeks of training to the control group level, and the level of collagen III decreased to be less than the level of the control group.

Relationship between hyperuricemia and adverse events in patients aged 40 years or older receiving rheumatic aortic valve replacement / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the prognostic value of hyperuricemia for adverse events in patients >40 years old receiving valve replacement surgery for rheumatic aortic valve disease.</p><p><b>METHDS</b>Consecutive middle-aged and elderly patients receiving aortic valve replacement surgery for rheumatic aortic valve disease between March, 2009 and July, 2013 were recruited in this study. The patients were divided into hyperuricemic group and normouricemic group based on their serum levels of uric acid, and the clinical data and adverse events within 1 year after the surgery were compared between the 2 groups.</p><p><b>RESULTS</b>A total of 632 consecutive patients were recruited, including 381 patients with hyperuricemia and 251 with normouricemia. The in-hospital mortality rate was significantly higher in hyperuricemic group than in normouricemic group (7.6% vs 2.0%, P=0.002). Serum uric acid levels were negatively correlated with eGFR (r=-0.421, P<0.001) and positively correlated with C-reactive protein level (r=0.093, P=0.025). Multivariate analysis showed that hyperuricemia was independently associated with the in-hospital mortality (OR=3.07, 95%CI: 1.13-8.37, P=0.028) and mortality at 1 year after the surgery (HR=3.14, 95%CI: 1.30-7.62, P=0.011) after adjusting for potential risk factors including age, NYHA III-IV and postoperative acute kidney injury (AKI). Kaplan-Meier analysis showed that the cumulative rate of 1-year mortality after surgery was significantly higher in patients with hyperuricemia (Log-rank=11.73, P=0.001).</p><p><b>CONCLUSION</b>Hyperuricemia is a predictor of in-hospital and one-year mortality in middle-aged and elderly patients following aortic valve replacement surgery for rheumatic aortic valve disease.</p>

Comparison of different administration routes of saikosaponin in plasma pharmacokinetics and brain pharmacokinetics / 中国中药杂志

To evaluate the optimum administration routes of saikosaponin in the treatment of epilepsy by comparing the plasma pharmacokinetics and the brain pharmacokinetics after different administration routes of saikosaponin. After receiving saikosaponin in different administration routes, the mice were sacrificed to collect the blood and brain tissues. The acetonitrile and methanol (9∶1) were used to precipitate the plasma protein. The concentration of the SSa in mice plasma and brain was determined by UPLC-MS/MS, and the pharmacokinetic parameters, bioavailability, the brain targeting coefficient (Re) and the brain drug targeting index (DTI) were calculated with Kinetica software. The relative brain Re was 142.17% by intranasal administration, with DTI of 3.06, significantly higher than those by the injections; in addition, the brain DTI was 1.25 by gavage administration. The brain drug targeting of saikosaponin by intranasal administration was higher than that by injection and gavage administration, indicating the advantages of the intranasal administration on medicine absorption into the brain.

Pharmacokinetics of bullatine A in Aconitum brachypodum total alkaloids gel in transdermal delivery / 中国中药杂志

In this study, the changes of bullatine A in plasma and skin of mice with time in microemulsion gel and ordinary gel of Aconitum brachypodum total alkaloids were compared through UPLC-MS/MS, and their pharmacokinetic parameters were also compared and analyzed, to investigate the feasibility of microemulsion agent in the transdermal drug delivery. UPLC-MS/MS method for simultaneous determination of bullatine A in plasma and skin had high sensitivity and was in line with the pharmacokinetic study requirements for transdermal drug delivery. The main pharmacokinetic parameters for microemulsion gel in the plasma were as follows: Cmax=(37.62±14.31) μg•L⁻¹, Tmax=(3.40±1.34) h, AUC0-∞=(1 027.7±260) μg•L⁻¹•h⁻¹, MRT=(34.80±12.31) h, MRTlast=(10.68±0.57) h, t1/2=(23.11±9.20) h; main pharmacokinetic parameters for ordinary gel in the blood: Cmax=(52.23±15.90) μg•L⁻¹, Tmax=(4.00±0.00) h, AUC0-∞=(728.60±280.80) μg•L⁻¹•h⁻¹, MRT=(20.69±3.98) h, MRTlast=(9.34±0.42) h, t1/2=(14.69±3.15) h. The results showed that the microemulsion gel had more stable transdermal absorption, longer duration of action and higher bioavailability than ordinary gel, indicating that the microemulsion gel had a good and stable transdermal effect. There was no significant difference in bioavailability of bullatine A in skin between microemulsion gel and ordinary gel.

In vitro recovery rate of bullatine A microdialysis probe / 中国中药杂志

To establish UPLC-MS/MS method for determination of the recovery rate of bullatine A microdialysis probe. The concentration difference method(incremental method, decrement method) was used to measure in vitro recoveries, and the effects of perfusate pH value, flow rate, concentration, and temperature on the recovery rate were investigated to explore the feasibility of microdialysis for the pharmacokinetic study of bullatine A. The method of UPLC-MS/MS showed good linear relationship within the required range; the specificity, recovery rate and precision of chromatography met the requirements of microdialysis samples. There was no significant difference in the measured recovery rate between incremental method and decrement method. Under the same conditions, in vitro recovery rate of the probe was decreased with the increase of flow rate, and was significantly increased with the increase of temperature, but was independent of bullatine A concentrations around the probe. The results showed that, microdialysis technology can be used for the pharmacokinetic study of bullatine A, and retrodialysis method (decrement method) can be used for the determination of the in vivo recovery rate of bullatine A microdialysis.

Study on pharmacokinetics of geniposide in mice administrated by xingnaojing microemulsion and mPEG2000-PLA modified xingnaojing microemulsion / 中国中药杂志

An HPLC method for the determination of geniposide concentration in mouse plasma was developed and the pharmacokinetics after intranasal administration of Xingnaojing microemulsion (XNJ-M) and mPEG2000-PLA modified Xingnaojing microemulsion (XNJ-MM) were investigated. Eighty mice were treated by XNJ-M and XNJ-MM nasally. The plasma samples were collected at different times and the drug in samples was detected by HPLC. The pharmacokinetic parameters were calculated by the software of Kinetica. The pharmacokinetic parameters of geniposide of XNJ-M were C(max) (4.36 +/- 2.69) mg x L(-1), t(max) 1 min, MRT (29.73 +/- 4.54) min, AUC (53.63 +/- 14.03) mg x L(-1) x min. The pharmacokinetic parameters of geniposide of XNJ-MM were C(max) (9.75 +/- 4.14) mg x L(-1), t(max) 1 min, MRT(22.34 +/- 2.90) min, AUC (131.87 +/- 40.13) mg x L(-1) x min. Geniposide can be absorbed into blood in a higher degree after intranasal administration with XNJ-MM compared to XNJ-M, which maybe caused by its less irritating and more absorption.

Quality characteristics of human spermatozoa with hyaluronic acid receptors / 中华男科学杂志

<p><b>OBJECTIVE</b>To analyze the quality characteristics of human spermatozoa with hyaluronic acid (HA) receptors and search for a new indicator for the assessment of sperm quality.</p><p><b>METHODS</b>Using sperm-HA binding assay with HA-coated slides, we determined the binding rate of motile sperm with HA receptors and analyzed its correlation with routine semen parameters, sperm membrane function, sperm fertilizing function and diminished/arrested sperm maturation.</p><p><b>RESULTS</b>The motile sperm with HA binding sites in the acrosomal region showed significantly higher acrosomal integrity ([95.4 +/- 3.9]%) and mitochondrial membrane potential (MMP) ([97.8 +/- 2.1]%) than those in the initial semen ([68.8 +/- 6.2]% and [72.8 +/- 7.4]%) (P < 0.01). The sperm-HA binding scores were correlated mildly with many routine semen parameters (r = 0.195-0.268, P < 0.05), positively with the acrosome reaction level after ionophore challenge (r = 0.666, P < 0.01) and normal sperm morphology (r = 0.417, P < 0.01), and negatively with sperm nucleoprotein immaturation (r = -0.266, P < 0.01), DNA fragmentation (r = -0. 308, P < 0.01) and excessive residual cytoplasm (r = -0.218, P < 0.05).</p><p><b>CONCLUSION</b>Sperm with HA receptors in the acrosomal region exhibit significant advantages in plasma membrane structure, fertilizing potential and maturation. The sperm-HA binding assay, which is based on a relationship between sperm receptors for zona pellucida and HA, is likely to become a new independent indicator for assessing the multiple qualities of spermatozoa.</p>

Phosphorylated modification of genistein and their interaction with lysozyme / 药学学报

A series of genistein's phosphates were synthesized through a simplified Atherton-Todd reaction and the structures of the phosphates were determined by electrospray ionization mass spectrometry (ESI-MS) and NMR. In case of monophosporyl derivatives, NMR spectra show that substitutions occur at the 7-position of genistein but not at its 4' and 5-position. Moreover, the non-covalent complexes of lysozyme with the four new genistein phosphates were detected by ESI-MS.

Optimization of formulation matrix proportion and preparation technology of realgar floating tablets for gastric retention by uniform design and correlation analysis / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize the different components proportions of the Realgar floating tablets for gastric retention by uniform design and correlation analysis.</p><p><b>METHOD</b>With the different dosage of hydroxypropyl methyl cellulose (HPMC) as the tablets frame matrix, uniform design and correlation analysis were used to optimize the best component proportions of formula, and to measure the dissolution of the tablets in vitro.</p><p><b>RESULT</b>Dissolution of the tablets in vitro was conformed to the expectation of experiment. The drug-release mechanism was by diffusion and corrosion at the same time.</p><p><b>CONCLUSION</b>The Realgar floating tablets for gastric retention achieved the goal of design, which demand sustained release and safety.</p>

Interaction of bovine serum albumin with luteolin and apigenin / 药学学报

<p><b>AIM</b>To study the interaction mechanism of bovine serum albumin (BSA) with luteolin and apigenin.</p><p><b>METHODS</b>Fluorescence quenching method and non-radioactive energy transfer theory were used.</p><p><b>RESULTS</b>The binding constants at different temperature were determined and the quenching mechanism of them were suggested as static quenching. The transfer efficiency of energy and distance between BSA and luteolin or apigenin were investigated according to the mechanism of the Förster energy transference.</p><p><b>CONCLUSION</b>The interaction between them seems to be strong and the binding force were mainly hydrophobic force. B(3')-OH,B(4')-OH strengthened the interaction of flavonoids and BSA.</p>

Isolation and Culture of Mesenchymal Stem Cells from Human Umbilical Cord Blood in Vitro / 中国康复理论与实践

@#ObjectiveTo investigate the feasibility and optimal condition of isolation,purification and expansion of mesenchymal stem cells(MSCs) derived from human umbilical cord blood in vtro.MethodsHuman umbilical cord blood(HUCB) was collected from full term deliveries scheduled,all samples were obtained sterilely with 20 U/ml preservative free heparin.The cord mononuclear cells were isolated with lymphocyte separation medium(density 1.077 g/ml),purified and expanded with MesencultTM medium and acidic environment to produce adherent layer.The surface antigen expression of MSCs was detected with flow cytometry.ResultsThe HUCB-derived mononuclear cells,when seeded in specific medium,gave rise to adherent cells,which exhibited either an osteoclast or mesenchymal-like phenotype.After passage 3,these cells were able to be purified and expanded.6.6×105 primary MSCs reached a number of 9.9×10<>sup8 after 10 expanded passage.Flow cytometry showed that MSCs did not express antigens CD34,CD11a and CD11b,but express strongly CD29 and weakly CD71,which was identical to human bone marrow-derived MSCs.ConclusionMSCs in HUCB can be cultured and expanded in vitro,and could be a source of stem cells for experimental and clinical application.

A minimally invasive approach in the treatment of complicated anal fistula through spatium intermuscular of anal sphincter / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the optimal operative approach for the complicated anal fistula.</p><p><b>METHODS</b>One hundred and ninety-two cases with complicated anal fistula were randomly divided into minimally invasive operation group (through spatium intermuscular of anal sphincter) and fistula resection group. The operation time, bleeding time during and after operation, pain lasting time, healing time of incision, area of anal scar, anal malformation and function and post operative recurrence were observed and compared between the two groups.</p><p><b>RESULTS</b>Compared to those of fistula resection group, the operative time was (36.5+/- 15.3)min, bleeding time during and after operation (2.0+/- 0.5)d, postoperative pain lasting time (1.5+/- 0.5)d, healing time of incision (18.5+/- 5.5)d in minimally invasive operation group. All were shortened (P< 0.05), and the incidence of anal malformation (5.2%, P< 0.01) and partial anal incontinence (2.1%, P< 0.01) was lower. There was no significant difference in postoperative recurrence between the two groups.</p><p><b>CONCLUSIONS</b>The minimally invasive operation through spatium in termuscular of anal sphincter is superior to fistula resection on the management of complicated fistula.</p>