Advances in drug delivery strategies against Helicobacter pylori / 药学学报

italic>Helicobacter pylori (H. pylori) can cause a variety of digestive tract diseases, the serious may develop into gastric cancer. Nowadays, H. pylori infection rate exceeds 50%, and its eradication rate is declining due to the continuous increase of drug resistance, leading to the occurrence of plenty of stubborn infections, which seriously threaten human health. At present, it is difficult to achieve satisfactory curative effect by increasing the types of antibiotics combination or increasing their dose. In this review, the clinical treatments of H. pylori were introduced. Proceed from the characteristics and pathological background of H. pylori infection that makes H. pylori difficult to eradicate, the research advances of drug delivery strategies for improving H. pylori eradication rate were reviewed, such as strategies that could increase drug concentration in stomach (e.g. drug delivery systems with gastric acid-stabilized ability), increase drug concentration in H. pylori colonization sites (e.g. drug delivery systems with gastric retention or H. pylori targeted abilities), overcome H. pylori resistance (metal nanoparticles, anti-biofilm delivery systems), enhance host immune response (vaccine preparation) and so on. Novel drug delivery systems, such as cell membrane coating technology and phage therapy, are comparatively rare in the field of anti-H. pylori, but have broad application prospects. This review would provide reference for the development and application of therapeutic strategies to improve H. pylori eradication rate.

Quality Value Transmitting of Substance Benchmark of Xiebaisan / 中国实验方剂学杂志

Objective:Based on fingerprint， index component content and dry extract yield， a quality evaluation method for substance benchmark of Xiebaisan was established to study the key quality attributes， to explore the quantitative transfer relationship between decoction pieces and substance benchmark， and to preliminarily formulate the quality standard of substance benchmark of Xiebaisan. Method:The substance benchmark of Xiebaisan was prepared according to the records of ancient formulas， fingerprints of 15 batches of decoction pieces and substance benchmarks were collected by high performance liquid chromatography （HPLC） and the index components were determined with the mobile phase of acetonitrile-0.05% phosphoric acid solution for gradient elution. The dry extract yield， fingerprint similarity and transfer rate of index components were combined to study the quantity value transmitting. Result:Ten characteristic peaks were identified in fingerprint of the substance benchmark and two characteristic peaks from stir-fried Mori Cortex， four characteristic peaks from baked Lycii Cortex， four characteristic peaks from Glycyrrhizae Radix et Rhizoma Praeparata cum Melle. Mulberroside A， liquiritin and glycyrrhizic acid were used as index components for the determination， the contents of mulberroside A， liquiritin and glycyrrhizic acid in substance benchmark of Xiebaisan were 2.69%-4.26%， 0.09%-0.17% and 0.09%-0.16%， and their transfer rates were （31.37±4.14）%， （36.12±4.03）% and （12.25±0.88）%， respectively. The similarity of fingerprint of substance benchmarks was good， the fingerprint similarities of 14 batches of substance benchmarks and control fingerprint were >0.9. The dry extract yield of substance benchmark of Xiebaisan ranged from 8.09% to 11.29%. Conclusion:The established quality evaluation method of substance benchmark of Xiebaisan is scientific and reasonable， and the transfer process of decoction pieces to substance benchmarks is stable and controllable. The preliminary quality standard of the substance benchmark can provide basis and reference for the development of modern preparations of Xiebaisan in the future.

HPLC fingerprint of famous traditional formula Sanpian Decoction and quality value transmitting of Chuanxiong Rhizoma / 中国中药杂志

Famous traditional formula Sanpian Decoction(SPD)comes from Dialectical Records of Chen Shiduo of the Qing Dynasty,and ranks among 100 classic prescriptions of Classic Famous Traditional Formula catalogue(the First Batch). SPD was prepared according to Management Standards for Traditional Chinese Medicine Decoction Room in Medical Institutions. According to the polarity of different components in SPD,two HPLC fingerprints were established, in which six herbs, namely Chuanxiong Rhizoma, Paeoniae Randix Alba, Sinapis Semen, Glycyrrhizae Radix et Rhizoma, Pruni Semen, Angelicae Dahuricae Radix,are all reflected in the fingerprints; The dry extract rate, transfer rate and similarities of fingerprints were used as indicators to study the relationship between the quality value transmitting of medicinal herbs-decoction pieces-whole decoction of Chuanxiong Rhizoma. Experiment result shows that,the transfer rate of ferulic acid from medicinal herbs to decoction pieces is between 72.00% and 108.36%; the transfer rate of ferulic acid from decoction pieces to SPD is between 31.76% and 64.09%; the dry extract rate of the whole decoction is between 14.69% and 20.16%;The similarity range of fingerprint 1 of 15 batches of SPD is between 0.971 and 0.998, and the similarity range of fingerprint 2 is between 0.980 and 0.996. The established fingerprint has rich information,and the established quality evaluation method is suitable for the quality control of medicinal herbs-decoction pieces-whole decoction of Chuanxiong Rhizoma, which can provide a certain reference for developing the quality control evaluation method for formulated granules, famous formulae and other terminal products derived from traditional Chinese medicine decoction.

Effect of Recipes for Tonifying Kidney and Replenishing Qi on Learning and Memory-related Genes in Aged Rats / 中国实验方剂学杂志

Objective:To observe the effects of recipes for tonifying kidney and replenishing Qi, Zuoguiwan (ZG) and Yiqi Congming Tang(YQ) on memory capacity, expressions of learning and memory-related genes expression, and explore the changes in relevant epigenetic modification enzymes. Method:SD male rats with natural aging (24 months old) were used as animal models and randomly divided into aged control group, aged ZG group(12.12 g·kg-1), aged YQ group(10.18 g·kg-1), aged compound group(11.15 g·kg-1) and aged antagonist RU38486 group(5×10-3g·kg-1). Another 5 months old male SD rats were included as the young control group. Morris water maze method was used to observe the spatial learning and memory ability of the rats. The co-localizations of histone deacetylase 2 (HDAC2) and methyl CpG binding protein 2 (MeCP2) in hippocampus of rats in each group were observed by laser confocal microscope. The changes in expressions of glucocorticoid receptor (GR), synapsin1(Syn-1), HDAC2, and histone acetyltransferase 1(HAT1) proteins in hippocampus of each group were detected by Western blot, and mRNA expression of HDAC2 was detected by Real-time fluorescence quantitative polymerase Chain reaction (Real-time PCR). At the same time, the effects of ZG, YQ and compound decoction in alleviating the above-mentioned abnormal changes were observed. Result:Compared with the young control group(control group), the latency of the aged control group was significantly prolonged (PPPPPPPConclusion:ZG group, YQ group, and compound group can improve the spatial learning and memory abilities of aged rats by increasing the expression of learning-memory-associated protein GR and epigenetic modification enzyme HAT1, and reducing the expression of HDAC2 and the co-localization of HDAC2 protein and MeCP2 in the nucleus.

An in vitro method for preliminarily screening competing agents to enhance drug permeation in cyclodextrin inclusion complexes / 药学学报

Cyclodextrin can increase the solubility of poorly soluble drugs, but also decrease the permeability of poorly soluble drugs in inclusion complexes simultaneously, which partially or completely counteracts the contribution of improvement in solubility to the oral absorption of poorly soluble drugs. If a competing agent is added to the system to compete binding sites of cyclodextrins with drugs, drug permeability can be improved by increasing the concentration of free drugs in the inclusion complex system. In this paper, a rapid in vitro screening method for competing agents of cyclodextrin inclusion complex is proposed based on the principle that good drug permeability is in accord with good cell uptake. The equilibrium constants between drugs and hydroxypropyl-beta-cyclodextrin (HPCD) were determined by phase equilibrium solubility method. Cinnarizine (CN) with a high equilibrium constant was selected as a competing agent, coumarin 6 (C6) and 9-octadecyl berberine (BD) with smaller equilibrium constants were selected as model drugs. Both changes of solubility and uptake by Caco-2 and A549 cells of C6 and BD were investigated different concentrations of CN to the HPCD solution of C6 and BD. The results showed that the uptake of C6 and BD increased in a CN concentration-dependent manner, and the solubility of C6 and BD in HPCD solution decreased with the prolongation of equilibrium time. It might be due to increased free drug concentrations that resulted from the competition of CN for drug binding sites with HPCD. In our study, in vitro cell uptake method was firstly used to validate the ability of CN as a competing agent to increase drug permeability (cell uptake). This method can be used for preliminarily screening of competing agents for drug-cyclodextrin inclusion complexes.

Improving oral absorption of BCS Ⅱ drugs by increasing solubility: frequently overlooked permeability / 药学学报

BCS Ⅱ drugs are characterized by low solubility and high permeability. Improving their solubility is considered an important approach to improve its oral absorption. Recent strategies to increase the solubility of poorly-soluble drugs may unexpectedly result in greatly depressed permeability, ultimately leading to failure in improving oral absorption. Based on the mathematics of membrane permeability coefficient of a drug, the membrane/aqueous partition coefficient is dependent on the drug's solubility in the gastrointestinal milieu, suggesting a unique interplay between the solubility and permeability of the drug, and treating the one irrespectively of the other may be insufficient. When we focus on the increase of drug solubility and overlook the efficacy of drug permeability, the positive effect of increased solubility to drug oral absorption might be traded off by depressed permeability. To provide rational formulary designs, by optimizing excipients and evaluation, this review summarizes solubility- permeability interplay for different types of solubilizing techniques, such as cyclodextrin, surfactants-based vehicle, cosolvent, amorphous solid dispersions, other infectors such as P-gp transporters and new techniques for simultaneous evaluation of drug solubility and permeability.

The curative effect and related influencing factors of hyperbaric oxygen combined with dexamethasone in the treatment of sudden deafness / 天津医药

Objective To analyze the curative effect and related influencing factors of hyperbaric oxygen combined with dexamethasone in the treatment of sudden deafness (SD). Methods The clinical data of 200 patients with SD, who were treated with hyperbaric oxygen combined with dexamethasone in our hospital from January 2014 to December 2016, were retrospectively analyzed.According to the curative effect,patients were divided into the effective group(n=182)and the ineffective group(n=18).Data of gender,age,time from onset to treatment,ears,hearing loss,the type of audiometric curve, dizziness, tinnitus, complications (hypertension or diabetes), plasma viscosity and serum C reactive protein (CRP) were compared between the two groups.Logistic regression analysis was used to screen risk factors influencing the curative effect of SD.Results The total response rate was 91.00%(182/200),and the ineffective rate was 9.00%(18/200).In the effective group patients aged > 50 years old, the time from onset to treatment was > 7 d, and the hearing loss was ≥ 60 dB. The proportions of high-frequency or stone-deaf audiometric curves, combined with dizziness, hypertension, diabetes, plasma viscosity ≥ 2 mPa·s and serum CRP level ≥ 20 μmol/L were significantly lower in the effective group than those in the ineffective group(P<0.05).Logistic regression analysis showed that age(>50 years old),time from onset to treatment(>7 d), hearing loss(≥60 dB), high-frequency or stone-deaf audiometric curves, combined with dizziness, hypertension, diabetes, plasma viscosity(≥2 mPa·s)and plasma CRP(≥20 μmol/L)were high risk factors for the curative effect of SD(P<0.05). Conclusion Hyperbaric oxygen combined with dexamethasone is an effective treatment for SD,but there are many factors affecting the curative effect. It is necessary to consider the clinical and pathological characteristics of patients in clinical treatment.

Bacterial biofilms: characteristics and combat strategies / 药学学报

Eighty percent of bacterial infections are related to the formation of bacterial biofilm. Compared with planktonic bacteria, bacterial biofilm is 10-1 000 times more resistant to antibiotics, which is the main cause of current bacterial drug resistance. A comprehensive understanding of the characteristics and resistance mechanisms of bacteria biofilm will help us treat the stubborn infections caused by the bacterial biofilm better and solve the problem of bacterial drug resistance. In this review, the composition and quorum sensing of bacterial biofilm, two major patterns of biofilm formation and drug resistance mechanisms were presented. Furthermore, representative compounds with anti-biofilm activity and compounds synergistic with antibiotics in anti-biofilm actions were introduced. Nano drug delivery strategies used for anti-biofilm in recent years as well as a novel drug delivery system-molecularly imprinted polymer was also introduced.

Effect of Over-Expression of KLF4 on the Proliferation and Apoptosis of K562 Cells In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effect of over-expression of KLF4 on the proliferation and apoptosis of K562 cells.</p><p><b>METHODS</b>The recombinant plasmid with over-expression of KLF4 was transfected into K562 cells by electroporation, then the obtained G418-resistant clones of K562 cells (K562/pEGFP-KLF4) were used as experimental group, and the vector control (K562/pEGFP-C1) and blank K562 cell control groups were set up at the same time. The expression of EGFP-KLF4 fusion protein in K562 cells was observed by fluorescence microscopy; the Western blot was performed to detect the level of KLF4 protein in the cells of each groups, the cell proliferation was tested by methylthiazolyl tetrazolium (MTT) assay, and the cell apoptosis was detected by flow cytometry.</p><p><b>RESULTS</b>Compared with K562/pEGFP-C1 group and blank K562 group, the level of KLF4 protein of K562/ pEGFP-KLF4 cells were significantly increased, and the over-expression ratio reached 74.07%(P<0.05). The proliferation ability of the over-expressing KLF4 K562 cells was inhibited significantly (P<0.05). However, the apoptosis of K562/pEGFP-ILK cells with over-expression of KLF4 was increased (P<0.05).</p><p><b>CONCLUSION</b>A stable over-expressing KLF4 protein K562 cell line with was constructed successfully, and the over-expression of KLF4 can inhibi proliferation and promote apoptosis of K562 cells.</p>

Differentiation Study of Chinese Medical Syndrome Typing for Diarrhea-predominant Irritable Bowel Syndrome Based on Information of Four Chinese Medical Diagnostic Methods and Brain-gut Peptides / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To establish discriminant functions of diarrhea-predominant irritable bowel syndrome (IBS-D) by studying it from quantitative diagnosis angle, hoping to reduce interference of subjective factors in diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.</p><p><b>METHODS</b>A Chinese medical clinical epidemiological survey was carried out in 439 IBS-D patients using Clinical Information Collection Table of IBS. Initial syndromes were obtained by cluster analysis. They were analyzed using step-by-step discrimination by taking information of four Chinese medical diagnostic methods and serum brain-gut peptides (BGP) as variables.</p><p><b>RESULTS</b>Clustering results were Gan stagnation Pi deficiency syndrome (GSPDS), Pi-Wei weakness syndrome (PWWS), Gan stagnation qi stasis syndrome (GSQSS), Pi-Shen yang deficiency syndrome (PSYDS), Pi-Wei damp-heat syndrome (PWDHS), cold-damp disturbing Pi syndrome (CDDPS). Of them, GSPDS was mostly often seen with effective percentage of 34. 2%, while CDDPS was the least often seen with effective percentage of 5.5%. A total of 5 discriminant functions for GSPDS, PWWS, GSQSS, PSYDS, and PWDHS were obtained by step-by-step dis- crimination method. The retrospective misjudgment rate was 4.1% (16/390), while the cross-validation misjudgment rate was 15.4% (60/390).</p><p><b>CONCLUSION</b>The establishment of discriminant functions is of value in objectively diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.</p>

Study on the Mechanism of Three Kinds Extracts of Qingxin Kaiqiao Recipe in Improving Learning and Memory Capabilities of AD Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the mechanism of three kinds extracts (saponins, volatile components, polysaccharide components) of Qingxin Kaiqiao Recipe (QKR) in improving learning and memory capabilities of Alzheimer's disease (AD) rats.</p><p><b>METHODS</b>A controlled comparison method was used. Totally 56 male SD rats were randomly divided into seven groups, i.e., the normal control group, the sham-operation group, the model group, the Aricept group, the saponin group, the benzene group, and the polysaccharide group, 8 in each group. AD rat model was established by bilateral hippocampus injection of Aβ1-40 (2 µL, 2.5 µg/µL). The next day after modeling rats in the saponin group, the benzene group, and the polysaccharide group, the saponin group, the Aricept group were intragastrically administered with saponin (at the daily dose of 9 mL/kg, 2.1 g/mL) , benzene (at the daily dose of 3.33 mL/kg, 5.7 g/mL) , polysaccharide (at the daily dose of 8.33 mL/kg, 2.28 g/mL), Aricept (at the daily dose of 1.67 mg/kg), respectively, once a day for 2 consecutive weeks from 10 am every day. Equal volume of normal saline was intragastrically administered to rats in the normal control group and the model group. Learning and memory capabilities were detected using water maze 2 weeks later. Expression levels of synaptotagmin-1 (Syt-1), interleukin-1β (IL-1β), glia fibrillary acidic protein (GFAP), and β-amyloid precursor protein (βAPP) in the cortex and hippocampus of AD rats were detected using immunohistochemistry.</p><p><b>RESULTS</b>Learning and memory capabilities could be improved by three kinds extracts of QKR. There was no statistical difference in the escape latency between the polysaccharide group and the model group (P >0. 05). The escape lacency was shortened in the rest treatment groups (P < 0.05). The escape latency was obviously prolonged in three kinds extracts of QKR groups, when compared with the Aricept group (P < 0.05, P < 0.01). Compared with the model group, times for crossing platforms were significantly increased in the saponin group and the Aricept group (P < 0.05). Compared with the Aricept group, average times for crossing platforms were significantly lessened in three kinds extracts of QKR groups (P < 0.01). Compared with the sham-operation group, expression levels of Syt-1, IL-1β, GFAP, and βAPP in the cortex and hippocampus were increased in the model group (P < 0.01). Compared with the model group, the expression of cortical Syt-1 increased in the saponin group and the benzene group; the expression of cortical IL-1β increased in the benzene group and the polysaccharide group; the expression of hippocampal GFAP increased in the three kinds extracts of QKR groups; expression levels of Syt-1, IL-1β, GFAP, and β-APP in the cortex and hippocampus decreased in the rest treatment groups (all P < 0.05, P < 0.01). Compared with the Aricept group, expression levels of Syt-1, IL-1β, GFAP, and βAPP in the cortex and hippocampus were significantly increased in three kinds extracts of QKR groups (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Three kinds extracts of QKR might play roles in anti-AD possibly by decreasing expression levels of Syt-1, IL-1β, GFAP, and βAPP in the cortex and hippocampus.</p>

Effects of qingxin kaiqiao recipe volatile oil on expressions of GFAP and caspase-3 in the cortex and hippocampus of AD rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study effects of Qingxin Kaiqiao Recipe (QKR) and its volatile oil on the expressions of Abeta(25-35) glial fibrillary acidic protein (GFAP), beta-amyloid (Abeta), beta-amyloid precursor protein (betaAPP), and Caspase-3 in the cortex and hippocampus of Alzheimer's disease (AD) rats induced by injecting Abeta(25-35) into the bilateral amygdala.</p><p><b>METHODS</b>Totally 32 male SD rats were selected. The AD rat model was establish by injecting Abeta(25-35) from bilateral amygdala. After modeling they were randomly divided into the model group, the Donepezil Hydrochloride group [Donepezil Hydrochloride Tablet (1.67 mg/kg), abbreviated as the DH group], the QKR group (QKR Decoction, 12.67 mL/kg), and the volatile oil group (3.33 mL/kg), 8 rats in each group. Another 8 rats were selected as the normal control group. Equal volume of double distilled water was administered to rats in the normal control group and the model group by gastrogavage, once daily for 2 successive weeks. The Morris water maze test was performed by the end of medication. The escape latency and times of crossing the platform in the water maze test were recorded during the 1st day to the fifth day. The expressions of GFAP, Abeta, betaAPP, and Caspase-3 in the cortex and hippocampus of the rats in each group were detected by immunohistochemical assay.</p><p><b>RESULTS</b>Compared with the model group, the escape latency from the 3rd day to the 5th day was shortened, the expressions of GFAP, Abeta, betaAPP, and Caspase-3 decreased in the cortex and hippocampus, the times of crossing the platform increased in each medication group, showing statistical difference (P < 0.01, P < 0.05). Compared with the DH group, the expressions of Abeta in the cortex and hippocampus decreased, and the betaAPP expression increased in the QKR group. The expressions of GFAP, betaAPP, and Caspase-3 in the cortex and hippocampus increased in the volatile oil group. The escape latency from the 3rd day to the 5th day was obviously prolonged, and the times of crossing the platform decreased in the volatile oil group, showing statistical difference (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>QKR could obviously improve the learning and memory capabilities of AD rats, which might be achieved through decreasing the expressions of GFAP, Abeta, betaAPP, and Caspase-3 in the cortex and hippocampus.</p>

Protective mechanism of cerebrospinal fluid containing qingxin kaiqiao recipe on PC12 cell injury induced by glutamate / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao recipe on PC12 cell injury induced by glutamate (Glu), in order to provide basis for the conical application of the recipe.</p><p><b>METHOD</b>SD rats were orally administered with decoction of Qingxin Kaiqiao recipe (7.9 g x kg(-1)) for three and a half days, 2 times a day, in order to prepare cerebrospinal fluid containing Qingxin Kaiqiao recipe. PC cells were divided into the normal group, the model group, the nimodipine group, the 10% normal CSF group, the 10% medicated CSF group, the 20% normal CSF group, the 20% medicated CSF group. Except for the normal group, other groups were cultured with PC12 cells and Glu with the final concentration of 20 mmol x L(-1) to establish the nerve cell injury model. Apart from the model group and the normal group, other groups were intervened with nimodipine, normal cerebrospinal fluid, and 10% and 20% medicated CSF. RT-PCR was used to detect the expression level of Bax mRNA, Bcl-2 mRNA and Caspase-3 mRNA, and MTT method was used to detect the activity of PC12 cells.</p><p><b>RESULT</b>The activity of PC12 cells of all of medicated CSF groups was higher than that of the model group, with the decrease in the expression of Bax mRNA and Caspase-3 mRNA and the increase in the expression of Bcl-2 mRNA. They showed a significant different with the model group (P < 0.01). The 20% medicated CSF group was superior than the 10% medicated CSF group (P < 0.01).</p><p><b>CONCLUSION</b>Qingxin Kaiqiao recipe shows an apparent protective effect on PC12 cells injured by Glu.</p>

Study on protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite-induced PC12 cell injury / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite (Na2S2O4)-induced PC12 cell injury, in order to provide basis for clinical application of the prescription.</p><p><b>METHOD</b>SD rats were orally administered with water decoction of Qingxin Kaiqiao Fang (7. 9 g . kg-1) once every 12 h, for a total of 7 times, in order to prepare cerebrospinal fluid containing Qingxin Kaiqiao Fang. The neurocyte injury model was established by adding Na2S2O4 with the final concentration of 8 m mol . L-1 into PC12 cells. With nimodipine (1 x 10(7)mol . L-1 ) as the positive control group, MTT method test was adopted to detect the impact of cerebrospinal fluid containing Qingxin Kaiqiao Fang on the activity of PC12 cells. The expression of Bax, Bel-2 and Caspase-3 mRNA was detected by RT-PCR.</p><p><b>RESULT</b>The cerebrospinal fluid containing Qingxin Kaiqiao Fang groups showed a significantly higher activity in PC12 cells than the model group, with decrease in expressions of Bax mRNA and Caspase-3 mRNA and increase in expression of Bel-2 mRNA. There were significant differences compared with the model group (P< 0. 05,P <0. 01).</p><p><b>CONCLUSION</b>Qingxin Kaiqiao Fang shows a notable protective effect on Na2S2 04-induced neurocyte injury.</p>

Protective effect of qingxin kaiqiao fang containing cerebrospinal fluid on PC12 cell injury induced by Abeta25-35 / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the protective effect of Qingxin Kaiqiao Fang containing cerebrospinal fluid on PC12 cell injury induced by Abeta25-35, in order to provide basis for clinical application of the formula.</p><p><b>METHOD</b>Sprague Dawley rats were orally administration with Qingxin Kaiqiao Fang (7.9 g x kg(-1)) twice a day for 3.5 days to prepare Qingxin Kaiqiao Fang containing cerebrospinal fluid. The nerve cell injury model was established by PC12 cells and Abeta25-35 with the concentration of 10 micromol x L(-1). The expressions of Bax, Bcl-2 and Caspase-3 mRNA were detected by immunohistochemical method in the PC12 cells.</p><p><b>RESULT</b>The Qingxin Kaiqiao Fang group showed a significant higher PC12 cell activity than the model group, with decrease in Bax mRNA and Caspase-3 mRNA expressions and increase in Bcl-2 mRNA expression. There was a significant difference from the model group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Qinxin Kaiqiao Fang shows a significant protective effect on Abeta25-35-induced nerve cell injury.</p>

Effect of qingxin kaiqiao recipe saponin on the expressions of Bax, Bcl-2, Abeta, and betaAPP in the cortex and hippocampus of Alzheimer's disease rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of Qingxin Kaiqiao Recipe (QKR) saponin on the expressions of Bax, Bcl-2, beta-amyloid (Abeta), and beta-amyloid precursor protein (betaAPP) in the cortex and hippocampus of Alzheimer's disease (AD) rats.</p><p><b>METHODS</b>Thirty-two SD rats of SPF grade were selected. Abeta 25 - 35 was injected into the bilateral amygdala to prepare the AD model. After modeling rats were randomly divided into the model group, the donepezil hydrochloride group (Donepezil Hydrochloride Tablet, 1.67 mg/kg), the QLR group (QLR Decoction, 12.67 mL/kg), and the saponin group (saponin, 6.30 mg/kg), 8 rats in each group. Another 8 rats were selected as the normal group. Rats in the normal group and the model group were given with equal volume of double distilled water by gastrogavage. The intervention was performed once daily for 2 successive weeks. The Morris water maze test was carried out by the end of medication. The escape latency and the platform crossing times were recorded during the 1 -5 days. The expressions of Bax, Bcl-2, Abeta, and betaAPP in the cortex and hippocampus were detected using immunohistochemical assay.</p><p><b>RESULTS</b>Compared with the model group, the 3rd - 5th day escape latency were all shortened in each medication group. The expressions of Bax, Abeta, and betaAPP decreased in the cortex and hippocampus. The numbers of platform crossing increased. The expression of Bcl-2 in the cortex increased. The expression of Bcl-2 in the hippocampus increased in the donepezil hydrochloride group and the QLR group with statistical difference (P<0.01, P<0.05). Compared with the donepezil hydrochloride group, the expression of betaAPP increased in the cortex and hippocampus of the saponin group. The expression of Abeta in the cortex and hippocampus decreased, the expression of Bax in the hippocampus decreased, and the expression of Bcl-2 in the cortex increased in the QLR group. The escape latency was obviously postponed at day 3 -5, the platform crossing times decreased, the expression of Bcl-2 in the cortex and hippocampus decreased in the saponin group, showing statistical difference (P<0.05, P<0.01).</p><p><b>CONCLUSION</b>QKR could significantly improve AD rats' learning, memory, and spatial capabilities, which might be achieved through decreasing the expressions of Bax, Abeta, and betaAPP in the cerebral cortex and hippocampus, and elevating the expression of Bcl-2.</p>

Effect of qingxin kaiqiao formula and saponin on learning and memory abilities and expression of apoptosis signal transducers Abeta and betaAPP in AD rat brain / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of qingxin kaiqiao formula and saponin on the learning and memory ability and the expression of the apoptosis signal transducers Abeta and betaAPP in AD rat brain.</p><p><b>METHOD</b>The comparative observation method was adopted for the animal test. Forty male SD rats were randomly divided into five groups, namely the normal group, the model group, the aricept group, the qingxin kaiqiao formula group and the saponin group, with eight rats in each group. Abeta(25-35) (10 g x L(-1)) was injected into their bilateral amygdala to establish the AD rat model. Since the next day, they were intragastrically administered with Aricept (1.67 mg x kg(-1)), Qingxin Kaiqiao decoction (12.67 mL x kg(-1)), saponin (6.30 mg x kg(-1)) and double distilled water filling for 2 weeks to observe their spatial memory ability in a Morris water maze and study the expression of Caspase-3, Abeta and betaAPP in brain tissues by immunohistochemistry.</p><p><b>RESULT</b>Each traditional Chinese medicine groups showed significant improvement in the learning and memory ability of AD rats and notable differences (P < 0.05, P < 0.01) compared with the control group. The qingxin kaiqiao formula group and the saponin group showed a decrease in the expressions of Caspase-3, Abeta and betaAPP in cerebral cortex and hippocampus area, displaying notable differences (P < 0.01, P < 0.05) compared with the control group.</p><p><b>CONCLUSION</b>qingxin kaiqiao formula and saponin can obviously improve the learning and memory ability of AD rats with by decreasing the expression of Caspase-3, Abeta and betaAPP in cortex and hippocampus.</p>

Effect of alloxan intraventricular injection on learning and memory abilities of mice / 中华神经医学杂志

Objective To investigate the effect of glucose metabolism alteration induced by alloxan intraventricular injection on learning and memory abilities of mice, and its role in the development of AD. Methods Mice were randomly divided into high-dose alloxan intraventricular injection group (n=7, 4 mg/kg) and low-dose alloxan intraventricular injection group (n=7, 1.5 mg/kg)and control group (n=7, physiological saline); intraventricular injection of alloxan, the O-GLcNAc transferase inhibitor, was performed in the high-dose and low-dose alloxan intraventricular injection groups to interfere the brain glucose metabolism. Morris water maze was used to detect the learning and memory abilities of mice. Western blotting and immunohistochemistry were used to analyze the alterations of phosphorylation and O-Glycosylation of neurofilament in mice brain induced by alloxan intraventricular injection. Results In the located navigation tests, the swimming time and distance to find the platform in the mice of alloxan administration were significantly increased as compared with those in the control group (P＜ 0.05); in space exploration experiments, compared with those in the control mice, the number of crossing the hidden platform was decreased and the initial angle of entry to water was increased in the mice of alloxan administration (P＜0.05). Western blotting and immunohistochemistry displayed that phosphorylation was obviously increased and the O-Glycosylation was significantly reduced in the cytoskletal neurofilament of the mice with alloxan administration as compared with those in the control group (P＜0.05), which was similar to the alteration of neurofilament's modification in AD brain. Conclusion The inventricular injection of alloxan could impair the learning and memory of mice, which might have a relation with the dysregulation of phosphorylation and O-Glycosylation in neurofilament caused by the impaired glucose metabolism, which is similar to the alteration of phosphorylation and O-Glycosylation in neurofilament in AD brain.

Simultaneous determination of serum concentrations of amoxicillin and clavulanate potassium in human blood using high-performance liquid chromatography / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a chromatography-based method for simultaneous analysis of the concentrations of amoxicillin and clavulanate potassium in human blood.</p><p><b>METHODS</b>With paracetamol as the internal control, human plasma samples, after treatment with methanol for protein sedimentation and centrifugation, were loaded for analysis with high-performance liquid chromatography (HPLC). HPLC analysis was carried out using a C18 column (5 µm, 4.6 mm×150 mm) with the mobile phase of acetonitrile-PBS (0.05 mol/L) of 10:90 (pH 2.3), UV detection wavelength of 220 nm, flow rate of 1.0 ml/min, and column temperature of 25 degrees celsius;.</p><p><b>RESULTS</b>The retention time of acetaminophen for potassium clavulanate, amoxicillin sodium and the internal control was 5.3, 7.2, and 8.5 min, respectively, and no interference by the endogenous impurities in the plasma samples was found. Amoxicillin sodium showed a good linearity within the concentration range of 0.52-4.16 µg/ml (r(2)=0.9996), and potassium clavulanate had a good linearity within the range of 0.266-2.14 µg/ml (r(2)=0.9998). The minimum detectable concentrations of amoxicillin sodium and potassium clavulanate were 0.065 µg/ml and 0.066 µg/ml, respectively. The relative recoveries of amoxicillin sodium were 95.9%-96.5% (n=5), and those of clavulanate potassium were 92.5%-98.8% (n=5); the intra- and inter-day RSD of amoxicillin sodium was 1.84%-6.4% and 2.1%-7.8%, as compared to that of potassium clavulanate of 3.57%-8.6% and 1.8%-9.1%, respectively.</p><p><b>CONCLUSION</b>This method is simple, accurate, sensitive, specific and reproducible for analyzing the concentrations of amoxicillin and clavulanate potassium simultaneously in human plasma.</p>

Eag-1 channel blocking inhibits the proliferation of glioma cells / 中华神经医学杂志

Objective To evaluate the influence of Eag-1 channel blocking on bioactivity of glioma cells in vitro. Methods Different small interfering RNAs (siRNAs) targeting for Eag-1 channel were designed and transfected to the U87 cells, and the blocking effects of those siRNAs were further confirmed on mRNA and protein levels by RT-PCR and Western blotting. The 50 nmol/l siRNAs (siRNA1 and siRNA2) and quinidine (5, 10, 20, 30 and 40 mmol/l) were used to block the activity of Eag-1 channel, respectively; and blank control group was also established. The proliferation of U87 cells 24, 48 and 72 h after the treatments was detected by MTT method; the changes of generation cycle,apoptosis ratio and intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Results High mRNA and protein levels of Eag-1 channel on glioma cell line U87 were confirmed in the blank control group, however, siRNA1 and siRNA2 transfection groups showed significantly lower mRNA and protein levels of Eag-1 channel on glioma cell line U87. MTT method indicated that, 24, 48 and 72 h after the treatments, the proliferation of U87 cells in the siRNA1 and siRNA2 transfection groups, and quinidine treatment groups (10, 20, 30 and 40 mmol/l) was significantly inhibited as compared with that in the blank control group (P＜0.05). The IC50 value of quinidine is33.7mmol/l. As compared with the blank control group, 50 nmol/L siRNA1 and siRNA2 transfection groups, and 33.7 mmol/l quinidine treatment group enjoyed a significantly increased cell percentage at G1 stage, cell apoptosis ratio and intracellular ROS level (P＜0.05). Conclusion Eag-1 channel blocking can obviously inhibit the proliferation of glioma cells, increase the cell percentage at G1 stage and intracellular ROS level, and induce apoptosis of glioma cells.