A novel peptide hydrogel of metal ion clusters for accelerating bone defect regeneration.

In the absence of adequate treatment, effective bone regeneration remains a great challenge. Exploring hydrogels with properties of excellent bioactivity, stability, non-immunogenicity, and commercialization is an important step to develop hydrogel-based bone regeneration materials. In this study, we engineered a self-assembled chelating peptide hydrogel loaded with an osteogenic metal ion cluster extracted from the processed pyritum decoction, including Fe2+, Cu2+, Zn2+, Mn2+, Mg2+, and Ca2+ ions, named processed pyritum hydrogel (PPH). We demonstrated that as a reservoir of beneficial metal ion clusters in bone regeneration, PPH has been shown to regulate a variety of genes in the process of bone regeneration. These genes are mainly involved in extracellular matrix synthesis, cell adhesion and migration, cytokine expression, antimicrobial and inflammation. Therefore, PPH accelerated the progress of various bone healing stages, and shortened the bone healing cycle by 4 weeks. Our investigation outcomes showed that the engineered metal ion cluster hydrogel is a novel, simple, and commercializable bone-regenerating hydrogel with potential clinical use.

Development of liquiritigenin-phospholipid complex with the enhanced oral bioavailability.

In the present study, liquiritigenin-phospholipid complex (LPC) was developed and evaluated to increase the oral bioavailability of liquiritigenin. A single-factor test methodology was applied to optimize the formulation and process for preparing LPC. The effects of solvent, drug concentration, reaction time, temperature and drug-to-phospholipid ratio on encapsulation efficiency were investigated. LPCs were characterized by UV-visible spectroscopy, differential scanning calorimetry (DSC), fourier transform infrared spectroscopy (FTIR), and powder X-ray diffractometry (PXRD). The apparent solubility and n-octanol/water partition coefficient were tested. The pharmacokinetic characteristics and bioavailability of the LPC were investigated after oral administration in rats in comparison with liquiritigenin alone. An LPC was successfully prepared. The optimum level of various parameters for liquiritigenin-phospholipid complex was obtained at the drug concentration of 8 mg·mL-1, reaction time for 15 min, reaction temperature of 30 ℃, a ratio of 1∶4.5 (W/W) drug-to-phospholipid and anhydrous ethanol as reaction solvent. Compared to liquiritigenin, the AUC0-t of the LPC was increased by 239%. The liquiritigenin-phospholipid complex significantly increase the lipid solubility and bioavailability of liquiritigenin, suggesting that it is an effective formulation for further development and clinical applications.

Screening for Potential Active Components of Fangji Huangqi Tang on the Treatment of Nephrotic Syndrome by Using Integrated Metabolomics Based on "Correlations Between Chemical and Metabolic Profiles".

As for traditional Chinese medicine (TCM) prescription, what puzzled researchers most was how to select proper chemical markers to represent the whole pharmacological action system. In this paper, an integrated metabolomic method was presented for a systematic discovery of potential active components in Fangji Huangqi Tang (FHT), a well-known TCM prescription for nephrotic syndrome treatment, based on "correlations between chemical and metabolic profiles." Firstly, a metabolomics study was carried out to select representative biomarkers of nephrotic syndrome. Then, after drug administration, the dynamic process of serum composition was investigated by the ultra-high performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry (UHPLC-ESI-Q-TOF-MS) technique to detect the prototypes and related metabolites of relative components from FHT. Pearson correlation analysis was finally used to find out the correlations between the endogenous metabolic spectrums and the chemical serum spectrums. As a result, 17 biomarkers for nephrotic syndrome indication were identified, and the main metabolic pathways of their concern included linoleic acid metabolism; cyanoamino acid metabolism; alpha-linolenic acid metabolism; glycine, serine, and threonine metabolism; arachidonic acid metabolism; and glycerophospholipid metabolism. Meanwhile, active components in FHT for nephrotic syndrome treatment were screened out, including (+)-tetrandrine demethylation, fenfangjine G hydrogenation, tetrandrine, N-methylfangchinoline, tetrandrine demethylation, fangchinoline, glycyrrhetic acid, astragaloside II alcohol dehydration, atractylenolide III demethylation + hydrogenation, atractylenolide III demethylation + hydrogenation, and licoricone-N-acetylcysteine conjugation. This study demonstrated a promising way to elucidate the active chemical material basis of TCM prescription.

[Problems and solutions in modern research of traditional Chinese herbal pieces processing technology].

Chinese medicine processing is the main feature that distinguishes traditional Chinese medicine from natural medicine and plant medicine, and is the main feature in clinical medication of traditional Chinese medicine. The research of Chinese medicine processing technology is an important link to realize standardization and standardization of Chinese herbal pieces, with urgent need to attract high attention. At present, there are still many problems in the research of processing technology of Chinese herbal pieces, mainly including inconsistent processing technology, large differences in process technology parameters, and unstable production technology of Chinese herbal pieces, resulting in uncontrollable quality of Chinese herbal pieces and affecting the clinical efficacy of Chinese medicine. This paper focused on the establishment of a unified standard processing technology, and put forward the countermeasures for the processing technology of Chinese medicine based on a comprehensive analysis of the current situations of the processing technology of Chinese herbal pieces, with significance for guiding the establishment of a standardized processing technology of Chinese medicine.

Effect of piperine on the bioavailability and pharmacokinetics of rosmarinic acid in rat plasma using UPLC-MS/MS.

1. The purpose of the present study was to investigate the effect of piperine (PP) on the pharmacokinetics of rosmarinic acid (RA) in rat plasma and to determine whether PP could enhance the oral bioavailability of RA via inhibition of its glucuronidation. 2. The pharmacokinetic profiles of RA between oral administration of RA (50 mg/kg) alone and in combination with different oral dose PP (20, 40, 60, and 80 mg/kg) to rats were investigated via a validated UPLC/MS/MS method. 3. The AUC and Cmax of RA were significantly increased in combination with different dose PP dose dependently, especially in the presence of 60 and 80 mg/kg PP (p < 0.01). The relative bioavailability of RA in the presence of 20, 40, 60, and 80 mg/kg PP was 1.24-, 1.32-, 2.02-, and 2.26-folds higher, respectively, compared with the control group given RA alone. Compared with RA, the pharmacokinetic modulations of RA glucuronide were even more apparent, and the glucuronidation of RA was remarkedly inhibited. 4. This study demonstrated that PP significantly improved the in vivo bioavailability of RA partly attributing to the inhibition of gut and hepatic metabolism enzymes of RA.

UHPLC-MS/MS method for the simultaneous quantitation of five anthraquinones and gallic acid in rat plasma after oral administration of prepared rhubarb decoction and its application to a pharmacokinetic study in normal and acute blood stasis rats.

Prepared rhubarb, as one of the main processed products of rhubarb, has a good effect on promoting blood circulation. In this paper we describe a rapid, sensitive, and selective ultra-fast liquid chromatography with tandem mass spectrometry method for simultaneous quantification of five anthraquinones (rhein, aloe-emodin, chrysophanol, emodin, and physcion) and gallic acid in plasma. Chromatographic separation was performed on an Extend C18 column at the temperature of 30°C using a mobile phase that consisted of 0.1% aqueous formic acid and acetonitrile. Satisfactory linearity, precision, accuracy, extraction recovery, and matrix effect have been achieved. Then, the validated method was successfully applied to a comparative pharmacokinetic study. The results might be helpful for guiding clinical application of prepared rhubarb in the future.

Combination of Morroniside and Diosgenin Prevents High Glucose-Induced Cardiomyocytes Apoptosis.

Cornus officinalis and Dioscorea opposita are two traditional Chinese medicines widely used in China for treating diabetes mellitus and its complications, such as diabetic cardiomyopathy. Morroniside (Mor) of Cornus officinalis and diosgenin (Dio) of Dioscorea opposita formed an innovative formula named M + D. The aims of the present study were to investigate myocardial protective effect of M + D on diabetic cardiomyopathy (DCM) through the inhibition of expression levels of caspase-3 protein, and identify the advantage of M + D compared with Mor, Dio, and the positive drug metformin (Met). We detected cell viability, cell apoptosis, intracellular reactive oxygen species (ROS) levels, and the expression levels of Bcl-2, Bax, and caspase-3 protein in rat cardiomyocytes. In result, Mor, Dio, and M + D increased cell viability, inhibited cell apoptosis and decreased ROS levels. Additionally, the expression of Bax and Bcl-2 protein was modulated and the expression levels of caspase-3 protein were markedly decreased. Among the treatment groups, M + D produced the most prominent effects. In conclusion, our data showed for the first time that Mor, Dio, and M + D prevented high glucose (HG)-induced myocardial injury by reducing oxidative stress and apoptosis in rat cardiomyocytes. Among all the groups, M + D produced the strongest effect, while Mor and Dio produced weaker effects.

Neuroprotective effects of Tongmai Yizhi Decoction () against Alzheimer's disease through attenuating cyclin-dependent kinase-5 expression.

OBJECTIVES: To explore the protective effects of Tongmai Yizhi Decoction (, TYD), a Chinese herb complex prescription against the impairment of cognitive functions and memory loss in amyloid beta 1-40 (Aß1-40) peptide and ibotenic (IBO)-induced Alzheimer's disease (AD) model rats. METHODS: The in vivo model was established by injecting Aß1-40 and IBO into left hippocampal CA1 area of Sprague-Dawley (SD) rat to mimic AD. Totally 32 SD rats were divided into 4 groups, including sham operation group, AD model group, TYD group [AD rats treated with TYD at the dosage of 19.44 g/(kg•d) for 4 weeks] and huperzine A group [AD rats treated with huperzine A at the dosage of 40.5 µg/(kg•d) for 4 weeks]. Spatial learning and memory level was detected by Morris Water Maze test. Histological morphology in the hippocampus was tested by hematoxylin-eosin (HE) staining. Cyclin-dependent kinase-5 (Cdk5) protein and gene expression level were investigated by Western blot analysis and real-time quantitative polymerase chain reaction (RT-qPCR), respectively. RESULTS: Aß1-40 and IBO treatment induced longer escape latency of rats, compared with sham operation group from day 25 (P<0.01). However, TYD and huperzine A obviously shortened the escape latency from day 26 (P<0.01). Moreover, the effect of TYD was similar to huperzine A (P>0.05). Furthermore, HE staining also showed that TYD and huperzine A reversed the neuropathological changes in the hippocampus triggered by Aß1-40 and IBO. TYD and huperzine A effectively reduced the expression levels of Cdk5 protein and gene located in rat hippocampus, compared with the AD model group (P<0.01). CONCLUSION: TYD could be a promising neuroprotective agent for protecting neuron from AD injury through inhibiting Cdk5 expression.

Evaluation of the Influence of Sulfur-Fumigated Paeoniae Radix Alba on the Quality of Si Wu Tang by Chromatographic and Chemometric Analysis.

An accurate and reliable method of high-performance liquid chromatographic fingerprint combining with multi-ingredient determination was developed and validated to evaluate the influence of sulfur-fumigated Paeoniae Radix Alba on the quality and chemical constituents of Si Wu Tang. Multivariate data analysis including hierarchical cluster analysis and principal component analysis, which integrated with high-performance liquid chromatographic fingerprint and multi-ingredient determination, was employed to evaluate Si Wu Tang in a more objective and scientific way. Interestingly, in this paper, a total of 37 and 36 peaks were marked as common peaks in ten batches of Si Wu Tang containing sun-dried Paeoniae Radix Alba and ten batches of Si Wu Tang containing sulfur-fumigated Paeoniae Radix Alba, respectively, which indicated the changed fingerprint profile of Si Wu Tang when containing sulfur-fumigated herb. Furthermore, the results of simultaneous determination for multiple ingredients showed that the contents of albiflorin and paeoniflorin decreased significantly (P < 0.01) and the contents of gallic acid and Z-ligustilide decreased to some extent at the same time when Si Wu Tang contained sulfur-fumigated Paeoniae Radix Alba. Therefore, sulfur-fumigation processing may have great influence on the quality of Chinese herbal prescription.

Chemical profiling of Qixue Shuangbu Tincture by ultra-performance liquid chromatography with electrospray ionization quadrupole-time-of-flight high-definition mass spectrometry (UPLC-QTOF/MS).

The present study was designed to develop and validate a sensitive and reliable ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) method to separate and identify the chemical constituents of Qixue Shuangbu Tincture (QXSBT), a classic traditional Chinese medicine (TCM) prescription. Under the optimized UPLC and QTOF/MS conditions, 56 components in QXSBT, including chalcones, triterpenoids, protopanaxatriol, flavones and flavanones were identified and tentatively characterized within a running time of 42 min. The components were identified by comparing the retention times, accurate mass, and mass spectrometric fragmentation characteristic ions, and matching empirical molecular formula with that of the published compounds. In conclusion, the established UPLC-QTOF/MS method was reliable for a rapid identification of complicated components in the TCM prescriptions.

Elemental Analysis of Nine Herbal Drugs for Nourishing Blood by Inductively Coupled Plasma Atomic Emission Spectroscopy with Microwave Digestion.

An analytical method has been developed for the determination of traces of Zn, Mn, Na, K, Ca, Mg, Ni, Pb, Se, Fe, Cu, Al, B, Ti, Sn, Hg, and Li in nine herbal drugs for nourishing blood by ICP-AES. The data showed that nine herbal drugs for nourishing blood were abundant in the contents of major and trace elements which are healthy for human body, including the trace elements with higher contents, such as Fe, Se, Ni, Mn, and Zn, which are well known for nourishing blood. Elements (macro-elements, essential elements, non-essential elements, and toxic elements) classified in nine herbal drugs for nourishing blood were illustrated in cylindrical histograms. Principal component analysis and hierarchical cluster analysis were applied to the data matrix to evaluate analytical results. The results of hierarchical cluster analysis showed that dendrogram obtained by Ward's method of hierarchical cluster analysis for seventeen variables and nine samples mainly displayed three clusters. After optimizing the microwave digestion technology, the recoveries of seventeen elements were in the ranges from 97.89% to 103.25% and the RSDs were all lower than 3.0%. ICP-AES combined with microwave digestion is accurate and precise in determining major and trace elements in nine herbal drugs for nourishing blood.

[A new strategy for exploring herb-pair investigation with a case study on Astragali Radix-Corni Fructus in treatment of diabetic nephropathy].

As the smallest unit of traditional Chinese medicinal formula compatibility, herb-pair has the basic characteristic of traditional Chinese medicinal formula compatibility. In recent years, herb-pair investigation has attracted much attention, which is an indispensable part of modernization of traditional Chinese medicine. With the decrease of the efficiency in the discovery of new drug, how to discover new drugs from traditional Chinese medicinal herb-pair has also been a bottleneck for the research and development of drug. The authors reviewed the domestic and foreign literatures in the latest years and summarized the current situations and the existing problems of herb-pair study. Based on these investigations, the authors innovatively proposed a novel concept of "precision herb-pair". Difference from traditional Chinese medicinal herb-pair or formulae with extensive roles and unclear efficacies, "precision herb-pair" belongs to a developed new mini herb-pair formula with an exact treatment and a relatively clear composition based on a certain specific disease. In addition, the authors also proposed a new strategy of "herb-pair - screen of multiple constituents based on column separation and in vitro cell viability - fuzzy target recognition pharmacology - re-evaluation of precision herb-pair", and successfully applied it to the development of a precision herb-pair from Astragali Radix-Corni Fructus in treatment of diabetic nephropathy. This proposed new strategy is simple, easy to carry out, and has a wide application, and can offer references and thoughts for the modern investigation of herb-pair and the research and development of new drug.

[Comparisons on chemical constituents of crude and wine-processed Dipsacus asper by using UPLC-Q-TOF/MS].

To compare the quality control indexes and chemical constituents of crude and wine-processed Dipsacus asper. According to Chinese Pharmacopoeia 2015 edition, water content, total ash, acid-insoluble ash and water soluble extract of different processed products were detected. UPLC-Q-TOF/MS approach was established to compare the contents of major constituents in crude and wine-processed D. asper. Moreover, the linearity, precision, stability, repeatability and recoveries of the approach were well studied. The results of water content, total ash, acid-insoluble ash and water-soluble extract of crude and wine-processed D. asper were all in line with the requirements of the Chinese Pharmacopoeia 2015 edition. Meanwhile, 20 main chemical constituents were identified by using UPLC-Q-TOF/MS. After wine-processing, the contents of asperosaponin Ⅵ, acetylate analogues and caffeic acid were significantly increased, while the contents of other phenolic components such as dicaffeoylquinic acid were decreased significantly, which may be which may be the main reason for different clinical efficacy of crude and wine-processed D. asper.

Evaluation of the influence of salt processing on pharmacokinetics of psoralen and isopsoralen in Psoralea corylifolia L.

A sensitive, specific and rapid ultra-high-pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method has been developed to investigate pharmacokinetic properties of psoralen and isopsoralen, two compounds isolated from raw/salt-processed fruit of Psoralea corylifolia L. UHPLC-MS/MS was used with positive ion electrospray. The mobile phase was composed of acetonitrile and 0.1% formic acid aqueous solution and a gradient elution program at flow rate of 0.3 mL/min was applied. Multiple reaction monitoring mode was used for the quantification of psoralen, isopsoralen ([M + H](+) m/z 187.0 → m/z 131.0) and scoparone (m/z 207.0 → m/z 151.1). Scoparone served as an internal standard. The method was fully validated for its sensitivity, selectivity, stability, matrix effect and extraction recovery. The obtained results showed that salt-processed Buguzhi significantly promoted the absorption of psoralen and isopsoralen, and increased the bioavailability of these compounds.

Dynamic accumulation analysis on bioactive constituents of Polygonum multiflorum in different collection periods.

To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.

Optimization of ultrasound-assisted extraction of phenolic compounds from Cimicifugae rhizoma with response surface methodology.

BACKGROUND: Cimicifugae rhizoma was a Ranunculaceae herb belonging to the composite family, and the roots of C. rhizoma have been widely used in tradition Chinese medicine. MATERIALS AND METHODS: Ultrasound-assisted extraction (UAE) of phenolic compounds from C. rhizoma. Caffeic acid (CA), isoferulic acid (IA), ferulic acid (FA), and total phenols were quantified by high-performance liquid chromatography-diode array detection and ultraviolet-visible spectrophotometer. Effects of several experimental parameters, such as ultrasonic power (W), extraction temperature (°C), and ethanol concentration (%) on extraction efficiencies of phenolic compounds from C. rhizoma were evaluated. RESULTS: The results showed that the optimal UAE condition was obtained with ultrasonic power of 377.35 W, extraction temperature of 70°C, and ethanol concentration of 58.37% for total phenols, and ultrasonic power of 318.28 W, extraction temperature of 59.65°C, and ethanol concentration of 64.43% for combination of CA, IA, FA. CONCLUSIONS: The experimental values under optimal conditions were in good consistent with the predicted values, which suggested UAE is more efficient for the extraction of phenolic compounds from plant materials.

Positive skeletal effect of two ingredients of Psoralea corylifolia L. on estrogen deficiency-induced osteoporosis and the possible mechanisms of action.

Estrogen replacement therapy (ERT) is utilized as a major regime for treatment of postmenopausal osteoporosis at present. However, long-term supplement of estrogen may cause uterine hyperplasia and hypertension leading to a high risk of endometrial cancer and breast cancer. Psoralea corylifolia L. has long been used as tonic and food additives in many countries. Previous studies had found two ingredients in P. corylifolia L.: bavachin and bakuchiol exhibited osteoblastic activity. The present study was designed to investigate the protective effect of bakuchiol and bavachin on ovariectomy-induced bone loss and explore the possible mechanism. In vivo, bakuchiol and bavachin could prevented estrogen deficiency-induced bone loss in ovariectomized rats without uterotrophic activity. In vitro studies suggested that bakuchiol and bavachin induced primary human osteoblast differentiation by up-regulating the Wnt signalling pathway. This study suggests that such a bone-protective role makes them a promising and safe estrogen supplement for the ERT.

[Effects of Wine-processing on Rhei Radix et Rhizoma on Upper-energizer Disease and Effects on Activities of Energy Metabolism Enzymes in Liver].

OBJECTIVE: To compare the effects of crude and wine-processed Rhei Radix et Rhizoma on upper-energizer disease and hepatic energy metabolism in mice. METHODS: The streptococcal pneumonia rats model and acetic acid burning mouth ulcers rats model were established and randomly divided into three groups: model group, crude Rhei Radix et Rhizoma group and wine-processed Rhei Radix et Rhizoma group. The pathologic changes were observed after the rats had been administrated with water extracts of crude and wine-processed Rhei Radix et Rhizoma respectively. The normal ICR mice were randomly divided into three groups: control group, crude Rhei Radix et Rhizoma group and wine-processed Rhei Radix et Rhizoma group. The influence of water extracts of crude and wine-processed Rhei Radix et Rhizoma on the activities of Na+, K-ATPase, Ca2+ -ATPase and succinic dehydrogenase(SDH) in the mice were compared. RESULTS: Compared with the crude one,the wine-processed Rhei Radix et Rhizoma significantly decreased the inflammation scores (P <0. 05), and promoted the tissue repair of acetic acid burning mouth ulcers rats model. The wine-processed one could also obviously reduce and normalize the level of leucocyte and neutrophilic granulocyte, lower the TNF-α level (P <0. 05), and relieve inflammatory exudation of the lung tissue. The inhibitory effects of wine-processed Rhei Radix et Rhizoma on the activities of SDH, Ca2+-ATPase and Na+, K + -ATPase were weaker than those of the crude one (P > 0. 05). CONCLUSION: After having been processed with wine, the efficacy of Rhei Radix et Rhizoma on upper-energizer disease is enhanced, and the inhibition on the activity of energy metabolism enzyme in liver tends to be weakened.

[Research status and prospect of primary processing of traditional Chinese medicinal materials].

The primary processing is important links and closely related to the quality of traditional Chinese medicinal materials, and is not only cleaning of remove the non-officinal parts, drying for termination the physiological status of organisms, but also retaining the most active substances, decreasing the toxic components, and promoting the transformation among chemical ingredients through primary processing. So the traditional primary processing endows with characters, quality, specifications and properties of traditional Chinese medicine, and embodies some important science truth. The traditional primary processing method and technology systems are derived from the long-term practices and experiences, which are distinctive, colorful, diverse, and scientific, which are helpful to development and utilization of traditional Chinese medicine resources. This paper systemically expounds the research status of the Chinese medicine processing method, summarizes the problems in the primary processing of traditional Chinese medicinal materials research, and prospects its bright future.

[Effect of emodin in attenuating endoplasmic reticulum stress of pancreatic acinar AR42J cells].

OBJECTIVE: To explore the effect of emodin on endoplasmic reticulum (ER) stress of pancreatic acinar AR42J cells. METHOD: Rat pancreatic acinar AR42J cells were cultured in 6-well plates, and divided into the normal control group, the model group (with the final concentration at 1 x 10(-7) mol · L(-1) for cerulean and lipopolysaccharide at 10 mg · L(-1)) and the emodin group (10, 20, 40 µmol · L(-1)). Cells in each group were cultured in three multiple pores for 24 h, and their supernate was removed after cell attachment. The normal control group was added with haploids, the model group was added with the modeling liquid for haploids, and the treatment groups were added with different concentrations of emodin at 15-20 min before the modeling liquid. The cells were continuously cultured for 3 h under 37 °C and 5% CO2. Their intracellular protease and lipase expressions were detected with kits. The cellular morphology was observed under optical microscope. The level of calcium in endoplasmic reticulum was measured under laser confocal microscopy. Western blot assay were used to determine the protein expression of ER-related signaling molecules. RESULT: Emodin could significantly inhibit levels of amylase, lipase and intracellular calcium and ER. CONCLUSION: Emodin could reduce pancreatic acinar cell injury induced by the combination of cerulean and lipopolysaccharide. Its action mechanism is correlated with the inhibition of intracellular calcium overload and ER stress.