Determination of seven ingredients of different grades Alismatis Rhizoma by QAMS method / 中国中药杂志

The present study is to establish a quantitative analysis of multi-components by single marker(QAMS) for determining contents of seven compositions in Alismatis Rhizoma, alismoxide, alisol C 23-acetate, alisol A, alismol, alisol B, alisol B 23-acetate and 11-deoxy-alisol B. Six relative correction factors(RCFs) of alismoxide, alisol C 23-acetate, alisol A, alismol, alisol B and 11-deoxy-alisol B were established in the UPLC method with alisol B 23-acetate as the internal standard, which was to calculate the mass fraction of each. The mass fraction of seven effective constituents in Alismatis Rhizoma was calculated by the external standard method(ESM) at the same time. Compared with the content results determined by the ESM and QAMS, the feasibility and accuracy of QAMS method were verified. Within the linear range, the RCFs of alismoxide, alisol C 23-acetate, alisol A, alismol, alisol B, 11-deoxy-alisol B were 0.946, 4.183, 0.915, 1.039, 0.923 and 1.244, respectively, with good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. Then, QAMS method was applied to determination of the different degree Alismatis Rhizoma from different areas. As a result, the concentrations of 7 components have differences in different areas, but no significant differences in different grades. The QAMS method is feasible and accurate for the determination of the seven chemical compositions, and which can be used for quality control of Alismatis Rhizoma.

Preparation of Rhus chinensis total phenolic acid pellets by extrusion-spheronisation method / 中国中药杂志

Extrusion-spheronisation method was used to prepare Rhus chinensis total phenolic acid pellets. The formula and preparation of R. chinensis total phenolic acid pellets were optimized. The formulas( drug loading capacity,diluent,wetting agent and anti-sticking agent) were determined by the single factor test with yield,appearance and performance as the indexes. The preparation was optimized by Box-Behnken design and response surface method,with the rate of extrusion,rate of spheronization and time of spheronization as the independent variables and the overall desirability value of yield,friability and roundness as the dependent variables. The optimal formula of pellets was as follows: drug loading capacity 28. 7%,MCC-lactose 9 ∶1,silicon dioxide as anti-sticking agent,and 60% ethanol as wetting agent. The optimal preparation was determined as follows: the rate of extrusion was 43 r·min-1,the rate of spheronization was 1 800 r·min-1,and the time of spheronization was 4 min. The absolute deviation between predicted value and estimated value under the conditions was less than 5. 0%,with a high degree of model fit. The preparation parameters obtained were accurate,reliable and reproducible. Under scanning electron microscopy( SEM),R. chinensis total phenolic acid pellets were uniform in diameter,round and smooth. The optimal formulation and process are stable and feasible for preparing R. chinensis total phenolic acid pellets.

Quality evaluation of Gualou Guizhi decoction based on chemical compositions and biological effects / 中国中药杂志

The paper was aimed to establish a quality evaluation model for Gualou Guizhi decoction based on the chemical compositions and biological effects. Ultra high performance liquid chromatograph-mass spectrometer was used to analyze and determine 24 kinds of chemical compositions in Gualou Guizhi decoction, and then, biological activity effect was quantitatively assessed in a zebrafish neuronal injury model which was induced by mycophenolate mofetil (MMF). As a result, the established method for quality evaluation of Gualou Guizhi decoction based on the chemical compositions and biological effects is feasible, stable and reliable, which can provide reference for quality control of compound Chinese medicines.

Screening of Lomefloxacin Aptamers Based on Polydopamine Nanospheres / 分析化学

Screening aptamers using nano-materials ( such as graphene oxide, gold nanoparticle, carbon nano-tube, etc. ) that can quench fluorescence and absorb single stranded DNA using hydrogen bond, π-πbond, charge transfer, and other non-covalent ways to combine with ssDNA, but without other conformational DNA, can excellently separate specific aptamers from non-specific ones. In this case, we can shorten the cycle numbers, enhance the success rate, and reduce the labour intensity of systematic evolution of ligands by exponential enrichment ( SELEX) . Especially for small molecular target, due to its difficulty in immobilization and small size, it is difficult to use traditional methods such as SPR-SELEX or affinity-SELEX for screening. In this experiment, polydopamine nanospheres ( MNPs@PDAs) were used to screen the Lomefloxacin. Also, we used magnetic separation technique to screen small molecular target rapidly. The interaction between aptamer candidates and the target could be monitored by recovery ratio of ssDNA and the whole MNPs@PDAs-SELEX process was performed through seven-round selection. As a result, we successfully obtained the aptamer named AF-3 which could recognize the lomefloxacin with high affinity (KD=(17. 57±0. 5) nmol/L). This screening method based on MNPs@PDAs makes it a promising reagent in the efficient aptamers selection of other targets.

Qualitative and quantitative analysis of major constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra by HPLC-DAD-Q-TOF-MS/MS / 中国中药杂志

In order to explore the differences of chemical constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra, a qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) was developed for identification of multi-constituents and an HPLC-DAD analytical method was developed for simultaneously determining 14 major compounds (gallic acid, protocatechuic acid, paeoniflorin sulfonate, protocatechuic aldehyde, methyl gallate, oxypaeoniflorin, catechin, albiflorin, and paeoniflorin, ethyl gallate, benzoic acid, pentagaloylglucose, benzoyl-paeoniflorin, and paeonol) in Paeoniae Radix Alba and Paeoniae Radix Rubra. Q-TOF/MS qualitative analysis was performed under negative ion mode and inferred 38 components of Paeoniae Radix Alba and 30 components of Paeoniae Radix Rubra. HPLC-DAD quantitative method result showed the contents of 8 ingredients were different between Paeoniae Radix Alba and Paeoniae Radix Rubra. The results indicated that the new approach was applicable in qualitative and quantitative quality control of Paeoniae Radix Alba and Paeoniae Radix Rubra.

Research on volatile components of gualou guizhi decoction and its components in rat brain by GC-MS / 中国中药杂志

To investigate the role of volatile components in the compound and to find the substance foundation of Gualou Guizhi decoction (GLGZD) for curing extremities spasticity after stroke. The chemical compositions of essential oil, obtained by hydrodistillation from Gualou Guizhi decoction and its major constituting herbs (Trichosanthis Radix, Paeoniae Alba Radix, Cinnamomi Ramulus, Zingiberis Recens Rhizoma, Glycyrrhizae Radix, Ziziphi Jujubae Fructus) were analyzed by GC-MS to evaluate the correlativity between volatile components of GLGZD and its major constituting herbs, and volatile components after oral administration of GLGZD in the rats' brain. Volatile components of GLGZD are mainly derived from Cinnamomi Ramulus, Zingiberis Recens Rhizoma, Ziziphi Jujubae Fructus, Trichosanthis Radix. The volatile components in the brain is mostly derived from radix trichosanthis. Compared with individual herbs of GLGZD, the dissolution of the components increase or new components appear after compatibility of six herbs. Adminstrated with GLGZD, the results point out that volatile components in the brain play a neuroprotective role through passing the brain.

Preparation of self-microemulsion drug delivery system of the mixture of paeonol and borneol based on Xingbi Fang / 药学学报

The aim of this study is to prepare self-microemulsifying drug delivery system (SMEDDS) of the mixture of paeonol (Pae) and borneol (Bor). Solubility test, ternary phase diagrams and simplex lattice method were employed to screen and optimize the formulation of the mixture of Pae and Bor-loaded SMEDDS. After formed into microemulsions, the particle diameter (PD) was determined and a TEM was employed to observe the microemulsions' morphology. The contents of Pae and Bor were determined by gas chromatography. As a result, while ethyl oleate (EO) as the oil phase, cremophor EL35 (EL35) as surfactant and Transcutol HP (HP) as cosurfactant, the range of the microemulsion on the ternary phase diagram was larger than other combinations. And at a ratio of 20:45:35, the microemulsions' PD was about 34 nm and the polydispersity index (PI) was about 0.2. There were 16% of Pae, 2% of Bor, 16% of EO, 37% of EL35 and 29% of HP in the prepared SMEDDS. The preparation process of the Pae and Bor-loaded SMEDDS based on Xingbi Fang is simple and feasible. This study provides a reference for the researches on the related traditional Chinese medicine and the related components.

Toxicity features of high glucose on endothelial cell cycle and protection by Dan Gua-Fang in ECV-304 in high glucose medium / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the toxicity features of high glucose on the endothelial cell cycle and the influence of Dan Gua-Fang, a Chinese herbal compound prescription, on the reproductive cycle of vascular endothelial cells cultivated under a high glucose condition; to reveal the partial mechanisms of Dan Gua-Fang in the prevention and treatment of endothelial injury caused by hyperglycemia in diabetes mellitus (DM); and offer a reference for dealing with the vascular complications of DM patients with long-term high blood glucose.</p><p><b>METHODS</b>Based on the previous 3-(4,5)-dimethylthiahiazo (z-y1)-3-5-diphenytetrazoliumromide (MTT) experiment, under different medium concentrations of glucose and Dangua liquor, the endothelial cells of vein-304 (ECV-304) were divided into 6 groups as follows: standard culture group (Group A, 5.56 mmol/L glucose); 1/300 herb-standard group (Group B); high glucose culture group (Group C, 16.67 mmol/L glucose); 1/150 herb-high glucose group (Group D); 1/300 herb-high glucose group (Group E); and 1/600 herb-high glucose group (Group F). The cell cycle was assayed using flow cytometry after cells were cultivated for 36, 72 and 108 h, respectively.</p><p><b>RESULTS</b>(1) The percentage of cells in the G0/G1 phase was significantly increased in Group C compared with that in Group A (P<0.05), while the percentage of S-phase (S%) cells in Group C was significantly reduced compared with Group A (P<0.05); the latter difference was dynamically related to the length of growing time of the endothelial cells in a high glucose environment. (2) The S% cells in Group A was decreased by 30.25% (from 40.23% to 28.06%) from 36 h to 72 h, and 12.33% (from 28.06% to 24.60%) from 72 h to 108 h; while in Group C, the corresponding decreases were 23.05% and 21.87%, respectively. The difference of S% cells between the two groups reached statistical significance at 108 h (P<0.05). (3) The percentage difference of cells in the G2/M phase between Group C and Group A was statistically significant at 72 h (P<0.01). (4) 1/300 Dan Gua-Fang completely reversed the harmful effect caused by 16.67 mmol/L high glucose on the cell cycle; moreover it did not disturb the cell cycle when the cell was cultivated in a glucose concentration of 5.56 mmol/L.</p><p><b>CONCLUSIONS</b>High glucose produces an independent impact on the cell cycle. Persistent blocking of the cell cycle and its arrest at the G0/G1 phase are toxic effects of high glucose on the endothelial cell cycle. The corresponding variation of the arrest appears in the S phase. 1/300 Dan Gua-Fang completely eliminates the blockage of high glucose on the endothelial cell cycle.</p>

Anticolchicine cytotoxicity enhanced by Dan Gua-Fang, a Chinese herb prescription in ECV304 in mediums / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of anticolchicine cytotoxicity of Dan Gua-Fang, a Chinesea Chinese), a Chinese herbal compound prescription on endothelial cells of vein (ECV304) cultivated in mediums of different glucose concentrations as well as the proliferation of those cells in the same conditions, in order to reveal the value of Dan Gua-Fang in preventing and treating endothelial damage caused by hyperglycemia in diabetes mellitus.</p><p><b>METHODS</b>The research was designed as three stages. The growing state and morphological changes were observed when ECV304 were cultivated in the culture mediums, which have different glucose concentrations with or without Dan Gua-Fang and at the same time with or without colchicine.</p><p><b>RESULTS</b>(1) Dan Gua-Fang at all concentrations reduced the floating cell population of ECV304 cultivated in hyperglycemia mediums. (2) Dan Gua-Fang at all concentrations and hyperglycemia both had a function of promoting "pseudopod-like" structure formation in cultivated ECV304, but the function was not superimposed in mediums containing both hyperglycemia and Dan Gua-Fang. (3) Colchicine reduced and even vanished the "pseudopod-like" structure of the endotheliocyte apparently cultivated in mediums of hyperglycemia or with Dan Gua-Fang. The "pseudopod-like" structure of the endotheliocyte emerged quickly in Dan Gua-Fang groups after colchicine was removed, but it was not the case in hyperglycemia only without Dan Gua-Fang groups. (4) Dan Gua-Fang reduced the mortality of cells cultivated in mediums containing colchicine. The cell revived to its normal state fast after colchicine was removed.</p><p><b>CONCLUSION</b>Dan Gua-Fang has the functions of promoting the formation of cytoskeleton and fighting against colchicine cytotoxicity.</p>

Effect of aike mixture on fibroplasia and inflammatory infiltration in rats with non-bacterial prostatitis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To assess the anti-inflammatory effect of Aike Mixture (AKM) on rats with nonbacterial prostatitis (NBP).</p><p><b>METHODS</b>Rat model of NBP was established by injection of Xiaozhiling Injection. The experimental rats were randomized into 7 groups: the three AKM groups (A1, A2 and A3), treated with high, middle and low dose of AKM respectively, the 2 positive control groups (C1 and C2), treated by Bazheng Mixture (BZM) and Qianliexian Decoction (QLXD) respectively, the model control group (Cm) and the sham-operative control group (Cso). The pathological changes in rats' prostate were observed by light microscope and transmission electron microscope.</p><p><b>RESULTS</b>Significant differences in the number and structure of acini, mesenchyma, as well as the degrees of anti-fibroplasia and inflammatory cell infiltration were shown between the treated groups (A1, A2, A3, C1, C1, C2) and the untreated groups (Cm, Cso), with statistical significance (P < 0.05 or P < 0.01). However, the anti-fibroplasia and anti-inflammatory effects in A1 were better than that in the two positive control groups significantly (P < 0.05 or P < 0.01).</p><p><b>CONCLUSIONS</b>AKM is a new TCM drug functioned for dispersing Gan-qi in treating NBP. It shows a better efficacy than that of BZM and QLXD, the two Chinese herbal medicines for clearing heat with remove dampness and activating blood circulation to remove stasis, respectively. Aike Mixture; prostatitis; pathological observation</p>

Regulatory T Cells and Their Molecular Markers in Peripheral Blood of the Patients with Systemic Lupus Erythematosus / 华中科技大学学报（医学）（英德文版）

Summary: CD4+CD25+ regulatory T cells (Tregs) and the expression of their molecular markers (GITR, Foxp3) in peripheral blood of the patients with systemic lupus erythematosus (SLE) were investigated in order to reveal the pathogenesis of SLE on the cellular and molecular levels. The level of Tregs in peripheral blood was detected by flow cytometry. The expression levels of GITR and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) were assayed by reverse transcriptasepolymerase chain reaction (RT-PCR). The level of IL-6 in the plasma was measured by ELISA.Comparisons were made among 3 groups: the active SLE group, the inactive SLE group, and normal control group. The level of Tregs in the active SLE group and the inactive SLE group was significantly lower than in the normal control group (P<0.01). The level of Tregs in the active group was lower than in the inactive group with the difference being not significant (P>0.05). The level of Tregs in SLE patients was significantly negatively correlated with the disease active index in SLE (SLEDAI) (r=--0.81, P<0.01). The expression levels of GITR mRNA in PBMCs of the active SLE group and the inactive SLE group were significantly higher than in the normal control group (P<0.05), and those of Foxp3 mRNA in SLE patients of both active and inactive SLE groups were significantly lower than in the normal control group (P<0.05). There was no significant difference in the expression of GITR and Foxp3 mRNA between the active SLE group and inactive SLE group (P>0.05). The plasma levels of IL-6 in both the inactive SLE group and active SLE group were significantly higher than in the normal control group (P<0.01). The plasma level of IL-6 in the active S LE group was significantly increased as compared with that in the inactive SLE group (P<0.05), and the plasma level of IL-6 in SLE was significantly positively correlated with SLEDAI scores (r=0.58, P<0.01) and significantly negatively correlated with the ratio of CD4+CD25+ cells/CD4+ cells (r=-0.389, P<0.05). It was concluded that the levels of Tregs and Foxp3 mRNA in peripheral blood of SLE patients were decreased and the levels of GITR mRNA and plasma IL-6 were increased. The Tregs and their molecular markers GITR, Foxp3 as well as the plasma IL-6 might play an important role in the pathogenesis of SLE.

Clinical study on treatment of chronic prostatitis/chronic pelvic pain syndrome by three different TCM principles / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To assess the efficacy of various therapeutic principles of TCM in treating patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS).</p><p><b>METHODS</b>Adopting blinded controlled trial method, 218 patients with CP/CPPS were randomly assigned to 4 groups: Group A treated with Aike Decoction for smoothing Gan-qi; Group B with Bazhengsan Decoction for clearing heat and removing dampness; Group C with Qianliexianyan Decoction for promoting blood circulation to remove stasis; and Group D with placebo. The scores of NIH chronic prostatitis symptom index (NIH-CPSI), clinical symptoms, including pain, symptoms of urinary tract and quality of life (QOL), and TCM syndrome integral were estimated at the beginning, the end of the 2nd and 4th week in the study.</p><p><b>RESULTS</b>Compared with the others, Group A showed a superiority in improving NIH-CPSI, scores of various clinical symptoms and TCM syndrome integral at the 2nd week, and improving NIH-CPSI, scores of pain and QOL at the 4th week (all P < 0.05), while the improvement on urinary tract symptoms and TCM syndrome integral in Group A at the 4th week were better than those in Group B and D, but insignificantly different to those in Group C, respectively. No adverse reaction occurred in Group A and D, but it did occur in the other two groups.</p><p><b>CONCLUSION</b>TCM therapy for smoothing Gan-qi shows good efficacy with quick initiating and high safety, it is an important principle for the treatment of CP/CPPS.</p>

Aike mixture has good anti-inflammatory and analgesic effects on mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the anti-inflammatory and analgesic actions of Aike Mixture (AKM).</p><p><b>METHODS</b>A total of 100 male mice were randomly assigned into 5 groups: a normal control group, a drug control group (a hydrocortisone subgroup and an atropine subgroup), a high-dose AKM group, a mid-dose AKM group and a low-dose AKM group. Xylene was spread on the left ear of the experimental mice to induce inflammation, and 1% acetic acid solution injected into the abdominal cavity to produce pain so as to cause the body bend. Different doses of AKM were given and their actions observed.</p><p><b>RESULTS</b>AKM had obvious anti-inflammatory effect on the xylene-induced ear tumefaction and inhibited the pain-caused body bend in the AKM groups, with significant difference from the normal control (P < 0.01).</p><p><b>CONCLUSION</b>AKM has good anti-inflammatory and analgesic effects, which is of clinical significance in the treatment of chronic prostatitis.</p>