Alkaline "Nanoswords" Coordinate Ferroptosis-like Bacterial Death for Antibiosis and Osseointegration.

Ferroptosis is an iron-dependent cell death and is associated with cancer therapy. Can it play a role in resistance of postoperative infection of implants, especially with an extracellular supplement of Fe ions in a non-cytotoxic dose? To answer this, "nanoswords" of Fe-doped titanite are fabricated on a Ti implant surface to resist bacterial invasion by a synergistic action of ferroptosis-like bacteria killing, proton disturbance, and physical puncture. The related antibiosis mechanism is explored by atomic force microscopy and genome sequencing. The nanoswords induce an increased local pH value, which not only weakens the proton motive force, reducing adenosine triphosphate synthesis of Staphylococcus aureus, but also decreases the membrane modulus, making the nanoswords distort and even puncture a bacterial membrane easily. Simultaneously, more Fe ions are taken by bacteria due to increased bacterial membrane permeability, resulting in ferroptosis-like death of bacteria, and this is demonstrated by intracellular iron enrichment, lipid peroxidation, and glutathione depletion. Interestingly, a microenvironment constructed by these nanoswords improves osteoblast behavior in vitro and bone regeneration in vivo. Overall, the nanoswords can induce ferroptosis-like bacterial death without cytotoxicity and have great promise in applications with clinical implants for outstanding antibiosis and biointegration performance.

Identification of JUN as determinant of osteoarthritis and its inhibition by the Chinese herbal formulae Zhuanggu Huoxue Tang.

Synovitis is an essential feature of Osteoarthritis (OA). Increasing evidence demonstrates that synovitis plays a critical role in OA's symptoms and structural progression. However, there is no effective drug for preventing and treating synovitis. Some Chinese herbal formulae have been found to treat clinical OA effectively, however, their mode of action is still unclear. This study investigated the Chinese herbal formulae Zhuanggu Huoxue Tang (ZHT) underlying mechanisms for treating osteoarthritis. Transcriptome data and a network pharmacology analysis were used to investigate the biochemical pathways affected by ZHT during OA treatment with in vitro verification. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were undertaken. The interaction network of the ZHT active constituent targets was determined using Cytoscape 3.7.1 software. Molecular docking of key pathogenic proteins and components of ZHT was performed in silico to confirm the compounds' pharmaceutical activities. The results establish that JUN is a target pathway in the pathogenesis of osteoarthritis. Miltirone, one of the active ingredients of ZHT, demonstrated a suitable binding activity with JUN. Miltirone alleviates the catabolic gene expression induced by IL-1ß and IL-6 in synovial fibroblasts (FLS), validating the use of Miltirone as a therapeutic drug for osteoarthritis.

Jujuboside A inhibits oxidative stress damage and enhances immunomodulatory capacity of human umbilical cord mesenchymal stem cells through up-regulating IDO expression.

Impaired immunomodulatory capacity and oxidative stress are the key factors limiting the effectiveness of mesenchymal stem cell transplantation therapy. The present study was aimed to investigate the effects of jujuboside A (JuA) on the protective effect and immunomodulatory capacity of human umbilical cord mesenchymal stem cells (hUC-MSCs). Hydrogen peroxide was used to establish an oxidative damage model of hUC-MSCs, while PBMCs isolated from rats were used to evaluate the effect of JuA pre-treatment on the immunomodulatory capacity of hUC-MSCs. Furthermore, Hoechst 33258 staining, lactate dehydrogenase test, measurement of malondialdehyde, Western blot, high-performance liquid chromatography; and flow cytometry were performed. Our results indicated that JuA (25 µmol·L-1) promoted the proliferation of hUC-MSCs, but did not affect the differentiating capability of these cells. JuA pre-treatment inhibited apoptosis, prevented oxidative damage, and up-regulated the protein expression of nuclear factor-erythroid factor 2-related factor 2 and heme oxygenase 1 in hUC-MSCs in which oxidative stress was induced with H2O2. In addition, JuA pre-treatment enhanced the inhibitory effect of hUC-MSCs against abnormally activated PBMCs, which was related to stimulation of the expression and activity of indoleamine 2,3-dioxygenase. In conclusion, our results demonstrate that JuA pre-treatment can enhance the survival and immunomodulatory ability through pathways related to oxidative stress, providing a new option for the improvement of hUC-MSCs in the clinical setting.

Active constituent of Polygala tenuifolia attenuates cognitive deficits by rescuing hippocampal neurogenesis in APP/PS1 transgenic mice.

BACKGROUND: Alzheimer's disease (AD) is the most common dementia worldwide, and there is still no satisfactory drug or therapeutic strategy. Polygala tenuifolia is a traditional Chinese medicine with multiple neuroprotective effects. In present study, we investigated the effects of three active constituents [3,6'-disinapoyl sucrose (DISS), onjisaponin B (OB) and tenuifolin (TEN)] of Polygala tenuifolia (PT) on the proliferation and differentiation of neural stem cells (NSCs) to identify the potential active constituent of PT promoting hippocampal neurogenesis. METHODS: NSCs were isolated from hippocampi of newborn C57BL/6 mice, and transfected with mutant amyloid precursor protein (APP) gene to establish an AD cell model (APP-NSCs). 3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays were performed, and the proliferation and differentiation of NSCs were assessed by neurosphere formation assay, 5-bromo-2'-deoxyuridine (BrdU) incorporation assay and immunofluorescence (IF) staining analysis. APP/PS1 transgenic mice were administrated with the potential active constituent DISS for 4 weeks. Morris water maze (MWM), Nissl staining assay and IF staining assays were carried out to evaluate the cognitive function, neural damages and hippocampal neurogenesis, respectively. RESULTS: DISS exerted the optimal ability to strengthen APP-NSCs proliferation and neuronal differentiation, followed by OB and TEN. Furthermore, DISS treatment for 4 weeks strikingly rescued the cognitive deficits, neuronal injures, and neurogenesis disorder in adult APP/PS1 transgenic mice. CONCLUSIONS: Our findings demonstrated that DISS is the constituent of PT that triggers the most potent increase of hippocampal neurogenesis in our mouse model of AD.

Noncanonical mono(ADP-ribosyl)ation of zinc finger SZF proteins counteracts ubiquitination for protein homeostasis in plant immunity.

Protein ADP-ribosylation is a reversible post-translational modification that transfers ADP-ribose from NAD+ onto acceptor proteins. Poly(ADP-ribosyl)ation (PARylation), catalyzed by poly(ADP-ribose) polymerases (PARPs) and poly(ADP-ribose) glycohydrolases (PARGs), which remove the modification, regulates diverse cellular processes. However, the chemistry and physiological functions of mono(ADP-ribosyl)ation (MARylation) remain elusive. Here, we report that Arabidopsis zinc finger proteins SZF1 and SZF2, key regulators of immune gene expression, are MARylated by the noncanonical ADP-ribosyltransferase SRO2. Immune elicitation promotes MARylation of SZF1/SZF2 via dissociation from PARG1, which has an unconventional activity in hydrolyzing both poly(ADP-ribose) and mono(ADP-ribose) from acceptor proteins. MARylation antagonizes polyubiquitination of SZF1 mediated by the SH3 domain-containing proteins SH3P1/SH3P2, thereby stabilizing SZF1 proteins. Our study uncovers a noncanonical ADP-ribosyltransferase mediating MARylation of immune regulators and underpins the molecular mechanism of maintaining protein homeostasis by the counter-regulation of ADP-ribosylation and polyubiquitination to ensure proper immune responses.

Icarisid II promotes proliferation and neuronal differentiation of neural stem cells via activating Wnt/ß-catenin signaling pathway.

Adult neurogenesis plays a vital role in maintaining cognitive functions in mammals and human beings. Mobilization of hippocampal neurogenesis has been regarded as a promising therapeutic approach to restore injured neurons in neurodegenerative diseases including Alzheimer's disease (AD). Icarisid II (ICS II), an active ingredient derived from Epimedii Folium, has been reported to exhibit multiple neuroprotective effects. In the present study, we investigated the effects of ICS II on the proliferation and differentiation of neural stem cells (NSCs) and amyloid precusor protein (APP)-overexpressing NSCs (APP-NSCs) in vitro. Our results demonstrated that ICS II dose-dependently suppressed apoptosis and elevated viability of APP-NSCs. ICS II (1 µM) potently promoted proliferation and neuronal differentiation of NSCs and APP-NSCs. ICS II (1 µM) significantly upregulated Wnt-3a expression, increased the phosphorylation of glycogen synthase kinase-3ß and enhanced the nuclear transfer of ß-catenin. Moreover, ICS II also promoted astrocytes to secrete Wnt-3a, which positively modulates Wnt/ß-catenin signaling pathway. These findings demonstrate that ICS II promotes NSCs proliferation and neuronal differentiation partly by activating the Wnt/ß-catenin signaling pathway.

Shenzao jiannao oral liquid, an herbal formula, ameliorates cognitive impairments by rescuing neuronal death and triggering endogenous neurogenesis in AD-like mice induced by a combination of Aß42 and scopolamine.

ETHNOPHARMACOLOGICAL RELEVANCE: According to the theory of traditional Chinese medicine (TCM), Alzheimer's disease (AD) is identified as "forgetfulness" or "dementia", and is mainly caused by "kidney essence deficiency" which ultimately induces "encephala reduction". Therefore, herbal formulas possessing the efficacy of nourishing kidney essence or replenishing brain marrow are commonly served as effective strategies for AD treatment. Shenzao jiannao oral liquid (SZJN), a traditional Chinese preparation approved by the China Food and Drug Administration (CFDA), is used for the treatment of insomnia and mind fatigue at present for its efficacy of nourishing kidneys. In present study, we found that SZJN could improve cognitive function of AD-like mice. AIMS OF STUDY: This study aims to investigate the effects of SJZN on ameliorating cognitive deficits of AD-like mouse model, and to illuminate the underlying mechanisms from the perspective of neuroprotection and neurogenesis. MATERIALS AND METHODS: Kunming mice (28 ± 2 g) were randomly allocated into seven groups: control, sham, model, donepezil and SZJN groups (low, middle and high). The AD mouse model was established by Aß42 combined with scopolamine. SZJN were intragastrically administrated at doses of 0.3, 1.5 and 7.5 g/kg for 28 days. Morris water maze (MWM) test was applied to determine the cognitive function. Hematoxylin eosin (HE) and Nissl staining were carried out to evaluate pathological damages in the cortex and hippocampal tissues. To explore the protective effects of SZJN on multiple pathogenic factors of AD, protein levels of Aß42, glial fibrillary acidic protein (GFAP), Bax, Bcl-2, Caspase-3, synaptophysin (SYP), brain-derived neurotrophic factor (BDNF), and neurogenesis related proteins were assessed using Immunofluorescence (IF) and western blot analysis. In vitro, the AD cell model was established by transduction of APP695swe genes into Neural stem cells (NSCs) isolated from the hippocampal tissues of neonatal C57BL/6 mice. Cell viability assay and neurosphere formation assay were carried out to verify the efficacy of SZJN on proliferation of NSCs. RESULTS: Our results demonstrated that SZJN (1.5 g/kg and 7.5 g/kg) treatment significantly ameliorated cognitive deficits of AD-like mice. SZJN (7.5 g/kg) treatment significantly retarded the pathological damages including neuronal degeneration, neuronal apoptosis, Aß peptides aggregation and reaction of astrocytes in AD-like mice. In addition, SZJN (7.5 g/kg) increased the expression of BDNF and SYP, and restored the abnormal level of MDA and SOD in the brain of AD-like mice. Furthermore, SZJN treatment for 28 days remarkably increased the proliferation of NSCs evidenced by more Nestin+ and BrdU+ cells in the hippocampal DG regions, and increased the amount of mature neurons marked by NeuN both in the cortex and hippocampal DG regions. In vitro, SZJN treatement (16, 32, 64 mg/ml) promoted the proliferation of NSCs evidenced by the increased amount and enlarged size of the neurospheres (p < 0.05). CONCLUSIONS: Our findings indicated that SZJN could ameliorate cognitive deficits by protecting neurons from death and triggering endogenous neurogenesis. Therefore, SZJN may be considered as a promising agent to restore neuronal loss and deter the deterioration in AD patients.

[Effect of Buyang Huanwu decoction (, BYHWD) on X-ray evaluation of early callus and level of serum alkaline phosphatase in elderly patients with Colles fracture after manual reduction and splint external fixation].

OBJECTIVE: To explore effects of Buyang Huanwu Decoction (, BYHWD) on early callus X-ray evaluation and level of serum alkaline phosphatase in elderly patients with Colles fracture after manual reduction and splint external fixation. METHODS: From October 2016 to October 2018, 60 elderly patients with Colles fractures were treated with manual reduction and splint external fixation and were divided into experimental group and control group. There were 30 patients in control group, including 15 males and 15 females; aged from 56 to 75 years old with an average of (67.81±5.41) years old; bone mineral density was (0.82±0.24) g/cm 2; patients were performed lift shoulders, bend and extend elbow joint, stretch five fingers and make a fist at 3 days after operation, 3 times daily for 1 month, 30 min once a time. There were 30 patients in experimental group, including 13 males and 17 females; aged from 57 to 77 years old with an average of (66.02±5.16) years old; bone mineral density was (0.76±0.23) g/cm2; patients performed rehabilitation exercise as control group and combined with BYHWD, 400 ml per dose, 2 times daily, 7 days as one course, totally 4 courses. RUSS scores at 14 and 28 days after reduction between two groups were compared, serum level of alkaline phosphatase (ALP) and serum calcium concentration were observed at immediately, 14 and 28 days after reduction. RESULTS: The patients between two groups were successfully fixed without re fractures and complications occurred. The patients were followed up for 30 to 35 days with an average of (31.60±1.03) days. RUSS score in experimental group at 14 and 28 days after reduction were 4.58±0.31 and 7.07±0.36, respectively; while in control group were 3.98±0.30 and 6.15±0.35, respectively; RUSS score in experimental group was significantly higher than that of control group. Serum alkaline phosphatase concentrations in experimental group at immediately, 14 and 28 days after reduction were (90.62±12.19) mmol/L ,(105.40±11.63) mmol/L, and (160.86±35.77) mmol/L respectively; while in controlgroup were (91.27±13.52) mmol/L ,(94.60±11.10) mmol/L ,(144.17±26.27) mmol/L respectively; there was no statistically difference between two groups at immediately; and had statistically differences between two groups at 14 and 28 days after reduction. There was no significant difference in serum calcium concentration between two groups at immediately, 14 and 28 days after reduction. CONCLUSION: BYHWD for elderly patients with Colles fracture could promote early formation of callus, effectively increase concentration of serum alkaline phosphatase and promote fracture healing.

Neuroprotective effects of ginseng protein on PI3K/Akt signaling pathway in the hippocampus of D-galactose/AlCl3 inducing rats model of Alzheimer's disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Alzheimer disease (AD) is a progressive neurodegenerative disease, with progressive memory loss, cognitive deterioration, and behavioral disorders. Ginseng (Panax ginseng C.A. Meyer) is widely used in China to treat various kinds of nervous system disorders. The study aimed to explore the therapeutic effect of ginseng protein (GP) on Alzheimer's disease and its correlation with the PI3K/Akt signaling pathway to understand the mechanism underlying the neuroprotective effect of ginseng. MATERIAL AND METHODS: The AD rat model was established by intraperitoneally injecting D-galactose [60mg/(kgd)] followed by intragastrically administering AlCl3 [40mg/(kgd)] for 90 days. From day 60, the GP groups were intragastrically administered with GP 0.05 or 0.1g/kg twice daily for 30 days. The ethology of rats was tested by Morris water maze test. The content of Aß1-42 and p-tau in the hippocampus of rats was detected by enzyme-linked immunosorbent assay. The expression of mRNAs and proteins of PI3K, Akt, phosphorylated Akt (p-Akt), Bcl-2, and Bax in the hippocampus was detected by real-time quantitative reverse transcription polymerase chain reaction and Western blot assay. RESULTS: GP was found to significantly improve the memory ability of AD rats and prolong the times of crossing the platform and the percentage of residence time in the original platform quadrant of spatial probe test. GP also reduced the content of Aß1-42 and p-tau and improved the mRNA and protein expression of PI3K, p-Akt/Akt, and Bcl-2/Bax in the hippocampus. CONCLUSIONS: GP could improve the memory ability and reduce the content of Aß1-42 and p-tau in AD rats. The anti-AD effects of GP were in part mediated by PI3K/Akt signaling pathway activation.

Gypenosides pre-treatment protects the brain against cerebral ischemia and increases neural stem cells/progenitors in the subventricular zone.

Gypenosides (GPs) have been reported to have neuroprotective effects in addition to other bioactivities. The protective activity of GPs during stroke and their effects on neural stem cells (NSCs) in the ischemic brain have not been fully elucidated. Here, we test the effects of GPs during stroke and on the NSCs within the subventricular zone (SVZ) of middle cerebral artery occlusion (MCAO) rats. Our results show that pre-treatment with GPs can reduce infarct volume and improve motor function following MCAO. Pre-treatment with GPs significantly increased the number of BrdU-positive cells in the ipsilateral and contralateral SVZ of MCAO rats. The proliferating cells in both sides of the SVZ were glial fibrillary acidic protein (GFAP)/nestin-positive type B cells and doublecortin (DCX)/nestin-positive type A cells. Our data indicate that GPs have neuroprotective effects during stroke which might be mediated through the enhancement of neurogenesis within the SVZ. These findings provide new evidence for a potential therapy involving GPs for the treatment of stroke.

Ocotillol Enhanced the Antitumor Activity of Doxorubicin via p53-Dependent Apoptosis.

The use of doxorubicin (Dox) was severely constrained by dose-dependent side effects, which might be attenuated by combining a "sensitizer" to decrease its cumulative dosage. In this study, it was investigated whether ocotillol could enhance the antiproliferation activity of Dox. MTT assays and xenograft tumor model were firstly conducted to evaluate the effect of ocotillol on the antitumor activity of Dox. Flow cytometry and Hoechst staining assays were then performed to assess cell apoptosis. Western blot and real-time PCR were finally used to detect the expression of p53 and its target genes. Our results showed ocotillol to enhance Dox-induced cell death in p53 wild-type cancer cells. Compared with Dox alone, Dox with ocotillol (Dox-O) could induce much more cell apoptosis and activate p53 to a much greater degree, which in turn markedly increased expression of proapoptosis genes. The enhanced cytotoxic activity was partially blocked by pifithrin- α , which might be through attenuating the increased apoptosis. Furthermore, ocotillol significantly increased the antitumor activity of Dox in A549 xenograft tumor in nude mice. These findings indicated that ocotillol could potentiate the cytotoxic effect of Dox through p53-dependent apoptosis and suggested that coadministration of ocotillol with Dox might be a potential therapeutic strategy.

Improved sedation for dental extraction by using video eyewear in conjunction with nitrous oxide: a randomized, controlled, cross-over clinical trial.

OBJECTIVE: The purpose of this study was to investigate the effect of a portable video eyewear entertainment system used in conjunction with nitrous oxide/oxygen sedation during the removal of impacted lower third molars. STUDY DESIGN: Thirty-eight patients had their bilateral third molars removed under local anesthesia and nitrous oxide/oxygen inhalation sedation in 2 visits. On one side, video eyewear was used (group NE). On the other side, the tooth was removed without the use of video eyewear (group N). Vital signs were monitored. Overall behavior and the outcome of treatment were assessed. RESULTS: All 38 patients completed the study. The mean scores on behavior rating in group NE were significantly higher than those in group N (P < .05). The majority of patients (92.1%) preferred nitrous oxide with video eyewear. CONCLUSIONS: The use of video eyewear appeared to augment the effectiveness of nitrous oxide sedation in dental extraction patients.

[Transurethral plasmakinetic enucleation of the prostate for benign prostatic hyperplasia].

OBJECTIVE: To evaluate clinical application of transurethral plasmakinetic enucleation of the prostate (PKEP) to the treatment of benign prostatic hyperplasia (BPH). METHODS: A total of 90 BPH patients, aged 59-83 (mean 71) years and with indication of surgery, underwent transurethral resection of the prostate (the TURP group, n=50) and transurethral plasmakinetic enucleation of the prostate (the PKEP group, n=40), respectively. We recorded and analyzed the preoperative prostate volume, IPSS, QOL and Qmax, operation time, intra- and post-operative bleeding and complications, postoperative continuous bladder irrigation, and IPSS, QOL and Qmax at 2 weeks and 6 months after surgery. RESULTS: The preoperative prostate volume and operation time were 58.9 g and 58.8 min in the TURP group versus 58.3 g and 93.0 min in the PKEP group. Mild transurethral resection syndrome (TURS) appeared in 2 TURP receivers, while no abnormality was found in electrocardiogram monitoring in those undergoing PKEP. Continuous bladder irrigation was necessitated in 3 and urgent incontinence of urine occurred in 4 cases of TURP, as compared with 1 and 4 cases in the PKEP group. None of the 90 patients needed blood transfusion. At 2 weeks before and after surgery and 6 months postoperatively, IPSS averaged 19.7, 11.6 and 5.1, QOL 4.6, 3.3 and 1.1, and Qmax 6.3, 13.0 and 18.1 ml/s in the TURP group versus 18.6, 8.4 and 4.9 (IPSS), 4.5, 2.7 and 1.1 (QOL) and 6.9, 14.2 and 19.0 ml/s (Qmax) in the PKEP group. There were significant differences in operation time, IPSS and QOL at 2 weeks postoperatively between the two groups, as well as in IPSS, QOL and Qmax at 6 months before and after surgery (P < 0.01). But no remarkable differences were found in preoperative prostate volume, IPSS, QOL and Qmax, 6-month postoperative IPSS and QOL, and Qmax at 2 weeks and 6 months after surgery between the two groups (P > 0.01). CONCLUSION: Transurethral PKEP is a safe, effective and thorough surgical method to be chosen for the treatment of BPH.

[Isolation and analysis of a new phytoecdysteroid from Cyanotis arachnoidea C. B. Clarke].

Cyanotis arachnoidea is a plant with plenty of phytoecdysteroid. To study the active compound in it, a new phytoecdysteroid with 5alpha-cholesta skeleton, was isolated from the whole plant of Cyanotis arachnoidea C. B. Clarke by using various chromatographic methods (alumina column chromatography, silica gel column chromatography, octadecyl silane (ODS) column chromatography, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC)). Its structure was analyzed on the basis of 1D and 2D nuclear magnetic resonances (NMR), electrospray ionization mass spectrometry (ESI-MS) methods. It is a compound with structure of 3beta,14alpha, 14alpha,20R,22R,25-hexahydroxy-5alpha-cholest-7-en-6-one, which is a rare phytoecdysteroid with 5alpha-H.

Progress on the screening and analysis of bioactive compounds in traditional chinese medicines by biological fingerprinting analysis.

Traditional Chinese medicines (TCMs), a key branch of natural medicines, play an important role in treatment of diseases because of their reliable clinical performance. Identification of their active compounds constitutes a bottleneck in the development of TCMs. Screening and analysis of active compounds is a challenge in TCMs research. This review summarizes recent progress in the development of biological fingerprinting strategies for screening and analyzing of bioactive compounds in TCMs using molecular recognition, metabolism and omics tools.The evaluated strategies including the following techniques: microdialysis/centrifugal ultrafiltration-HPLC, biochromatography, metabolic fingerprinting analysis, 2-dimensional biochromatography and omics fingerprinting analysis.

The effect of chemical sympathectomy and stress on bone remodeling in adult rats.

OBJECTIVES: Bone remodeling has recently been revealed to be under sympathetic nerve control. The role of the sympathetic nerve system is not clearly understood. The present study aim to explore the effect of chemical sympathectomy and stress on bone remodeling in adult rats. METHODS: 24 twelve-month-old Wistar rats were divided into three group (sympathectomy, stress and control). The sympathectomy and stress group rats were administered 6-hydroxydopamine (150 mg/kg each day) and saline (1 ml/kg each day) intraperitoneal respectively for one week and exposed to stress procedure for another three weeks. The stress procedure was mild, unpredictable footshock, administered for one hour once daily. Analysis of serum chemistry, microcomputed tomography, dual energy X-ray absorptiometry, biomechanical testing and bone histomorphometry were employed. RESULTS: The stress group rats showed increased bone resorption in contrast to the sympathectomy and control group rats. The serum level of calcium and phosphorus cations and norepinephrine were enhanced, the cancellous bone volume and bone mineral density were reduced, bone mechanical property such as strength, ductility and toughness were weakened, the osteoclast counts and osteoclast surfaces were increased and the bone formatin rate were decreased significantly in the stress group rats in contrast to the other two groups rats. There was no significant difference of bone remodeling between the sympathectomy group and control group rats. CONCLUSION: Our study showed stress-increased sympathetic nerve system activity enhanced bone resorption while chemical sympathectomy inhibited bone resorption under stress. We postulate sympathetic neurotransmitter and neuropepitide may play a role in regulating bone remodeling.

Evaluation of the interaction of bioactive compounds in Cortex Pseudolarix and Radix Stephaniae by the microdialysis probe coupled with high performance liquid chromatography-mass spectrometry.

An efficient and convenient method, biological fingerprinting chromatogram analysis is presented, which is applied to the comparison of fingerprinting chromatograms of the extracts of Chinese herbal medicines after the interaction with biological systems (cell, DNA, protein, etc.). The method was established for the purpose of screening and analysis of the multiple bioactive compounds in herbal medicines. In this work, microdialysis sampling combined with high performance liquid chromatography-mass spectrometry (HPLC-MS) was studied for binding property of MCF-7 and multidrug resistant MCF-7 cell systems. The results showed that pseudolaric acid A (PAA) and pseudolaric acid B (PAB) in the cortex of Pseudolarix kaempferi (Lamb.) Gorden can easily bind to the MCF-7 cells ranging from 0 to 16.3% (PAA) and from 0 to 35.7% (PAB), and another compound, tetrandrine (TET) from the root of Stephania tetrandrae S. Moore, showed higher binding activity with multidrug resistant MCF-7 cells ranging from 0 to 39.9%.

Comprehensive two-dimensional high-performance liquid chromatography system with immobilized liposome chromatography column and reversed-phase column for separation of complex traditional Chinese medicine Longdan Xiegan Decoction.

A comprehensive two-dimensional HPLC system with an immobilized liposome chromatography (ILC) column in conjunction with an RP column (in tandem) was developed for the screening and analysis of the membrane-permeable compounds in a traditional Chinese medicine prescription Longdan Xiegan Decoction (LXD). More than 50 components in LXD were resolved using the developed separation system. Eight flavonoids and two iridoids were identified interacting with the ILC column; a system that mimics biomembranes. The results show that the developed comprehensive two-dimensional chromatography system can be used for identifying membrane permeable natural products in complex matrixes such as extracts of traditional Chinese medicine prescriptions.

Biological fingerprinting analysis of the traditional Chinese prescription Longdan Xiegan Decoction by on/off-line comprehensive two-dimensional biochromatography.

A comprehensive two-dimensional biochromatography method using a silica-bonded human serum albumin (HSA) column and a RP-HPLC column was developed for the biological fingerprinting analysis of bioactive components in a traditional Chinese medicine (TCM) prescription, Longdan Xiegan Decoction (LXD). The biochromatography with HSA-immobilized stationary phase was applied to study the interaction of multiple components in LXD with HSA in the first dimension, and fractions of HSA column were further separated by a silica monolithic ODS column (on-line)/an ODS column (off-line) coupled with a diode array detector and an atmospheric pressure chemical ionization mass spectrometer (APCI-MS). More than 100 compounds in LXD that interacted with the immobilized HSA were separated and analyzed. Among them 19 compounds were identified based on their retention values, UV spectra, molecular weights and mass spectra. The results show that the developed comprehensive two-dimensional biochromatography system reported here is capable of being used for biological fingerprinting analysis of natural products in complex matrices such as extracts of TCMs and their prescriptions.

Affinity chromatography with immobilized DNA stationary phase for biological fingerprinting analysis of traditional Chinese medicines.

A stationary phase for high performance affinity chromatography with immobilization of DNA onto silica gel was prepared and characterized. The effect of the ionic strength, concentration of Mg2+, EDTA and CH3CN in the mobile phase on the retention of alkaloids were investigated. With this stationary phase, biological fingerprinting analysis of traditional Chinese medicines (TCMs) Coptis chinensis Franch and Rheum palmatum L. was performed with both one-dimensional (1-D) and two-dimensional (2-D) chromatography. The 1-D chromatography was performed with isocratic and gradient elution and 2-D chromatography was developed with immobilized DNA column combined with silica monolithic ODS column. It was found that 7 compounds in Coptis chinensis Franch including berberine, palmatine and jatrorrhizine, 14 compounds in Rheum palmatum L. including aloe-emodin, rhein, emodin, chrysophannol-8-O-glucophranoside and physionl-8-O-glucophranoside were active in binding to the immobilized DNA.