Metabolomics study of the effect of Danggui Buxue Tang on rats with chronic fatigue syndrome.

Danggui Buxue Tang (DBT), a traditional Chinese medicine (TCM) formula for 'invigorating qi and enriching blood', has been reported to produce a good effect on chronic fatigue syndrome (CFS). However, the related mechanism remains largely undetermined. This study devised a metabolomics approach with GC-MS combined with pattern recognition to estimate the extent to which DBT alleviated CFS induced by food restriction and force swimming in rats. After 4 weeks of treatment, the endurance capability of rats was significantly better, and the motionless time was significantly shorter in the DBT group than in the CFS model group. Moreover, the activities of superoxide dismutase and glutathione peroxidase increased, whereas the levels of malondialdehyde, interleukin 6, and tumor necrosis factor alpha decreased in the DBT treatment group. Fifteen significantly changed metabolites were observed in the serum of rats with CFS, which was reversed markedly by DBT treatment. Metabolic pathway analysis showed that DBT could possibly alleviate CFS in rats by regulating the biosynthesis of phenylalanine, tyrosine, and tryptophan and the metabolism of glycine, serine, threonine, glycerolipid, glyoxylate, dicarboxylate, and tyrosine. It was observed that the metabolism of glycine, serine, and threonine was most closely related to the improvement in CFS by DBT treatment. This study showed that DBT could improve CFS effectively, and metabolomics was a powerful means to gain insights into the TCM formulas against CFS.

Amidoxime functionalized chitosan for uranium sequestration in vivo.

Amidoxime functionalized chitosan (AC) was recommended as a chelator for uranium sequestration in vivo in this study, and the structure-activity relationship was also explored. Compared with ZnNa3-DTPA, which was a commercial uranium mobilization drug, AC exhibited excellent biocompatibility and uranium removal efficiency, whether by injection or orally, which could reduce the amounts of uranium deposited in kidneys and femurs by up to 43.6% and 32.3%. In particular, ACs still possessed the ability to mobilize uranium in vivo even if administration was delayed for 72 h.

A New Surgical Method to Reconstruct the Fingertip.

BACKGROUND: In our previous study, we used cross finger fascial flap combined with split-thickness toe nail bed graft to reconstruct large area defect of the nail bed with distal phalanx exposure. Fingertip avulsions are common injuries to the upper extremity and cause a great deal of distress for patients. Replantation, amputation and flap coverage are considered. However, all these methods have their limitations. METHODS: Five patients with fingertip avulsions were enrolled. We used the methods mentioned above combined with the reversed pedicled island flap to covered the defects of both the dorsal and volar sides and reconstruct the fingertip. RESULTS: All the 5 patients were followed for at least 3 months. The blood supply of the flaps and nail bed is good. All the patients were satisfied with their new fingertips. CONCLUSIONS: The method we introduced in this study is proved to be an effective surgical method for fingertip avulsions.

Metabonomic analysis of serum reveals antifatigue effects of Yi Guan Jian on fatigue mice using gas chromatography coupled to mass spectrometry.

Yi Guan Jian (YGJ), one of the most commonly used traditional Chinese medicines, has been reported to possess significant antifatigue effects. However, the mechanisms underlying its antifatigue effects remain largely unresolved. In this study, a metabonomics approach, involving gas chromatography coupled to mass spectrometry and a multivariate statistical technique, was developed to estimate the extent to which YGJ alleviated the exhausting swimming-induced fatigue of mice. High-dose treatment with YGJ significantly extended the swimming time of fatigued mice. Significant alterations of metabolites involving amino acids, organic acids and carbohydrates were observed in the serum of fatigued mice, which were reversed by YGJ treatment while biochemical indexes returned to normal. These metabolic changes suggest that the antifatigue effect of YGJ is associated with the impairement of amino acid, organic acids and carbohydrates. It also appears that YGJ can induce significant metabolic alterations independent of the exhausting swimming-induced metabolic changes. The significantly altered metabolites induced by YGJ intervention include l-2-amino-acetoacetate, taurine, fumaric acid, malic acid, oxoadipic acid and l-aspartate, all of which are associated with antifatigue properties. This suggests that YGJ exerts chemopreventive effects via antifatigue mechanisms.

[Anti-inflammation effect of Jinlingzi San in rat metabonomics based on 1H-NMR and LC-MS technology].

To further explore the regulatory effect of Jinlingzi San on in vivo inflammatory mechanism during inflammatory treatment, this study adopted 1H-NMR and LC-MS technology to analyze differences in in vivo metabolites of carrageen-induce rat foot swelling model. Besides, biomarkers related to inflammation models of Jinlingzi San in SD rats were discovered to speculate the regulatory mechanism of Jinlingzi San in resisting carrageen-induce inflammation. Through the analysis of detection spectrum, we found 18 biomarkers of metabolites(citrate, pyruvate, malic acid, succinate, glutamate, lysine, tartrate, 2-oxobutyric acid, glycine, guanosine, 9-cis-retinoic acid, triphosphate, inosine 5'-diphosphate, inosine diphosphate, tripolyphosphate, inorganic triphosphate, glycerophosphocholine, 21-deoxycortisol). Relevant pathway analysis results were TCA cycle, pyruvate metabolism, glycine, serine and threonine metabolism, and dicarboxylic acid metabolism. From the metabolic network, we can see that the anti-inflammatory effect of Jinlingzi San can regulate citric acid, succinic acid and glycine content to resist oxygen free radical and reduce body damage by ROS, so as to down-regulate inflammatory factors generated from body tissues and resist inflammation.

[Subacute toxicity metabonomics of Jinlingzi powder based on LC-MS].

To further understand the metabolic characteristics of Jinlingzi powder toxicity effect in rats and explore the effect of Jinlingzi powder on unknown biological pathways in the treatment process. In this experiment, the effect of three doses of Jinlingzi powder decoction on rat liver and kidney was investigated to explore the characteristics and rules of Jinlingzi powder on in vivo metabonomic changes in rats. First, urine and serum samples of the rats were used for LC-MS analysis. Under the XCMS online analysis, 44 differential substances were found in the identification of metabolites. Finally, Metpa was used for metabolic pathways enrichment and analysis, and five related metabolic pathways were obtained: steroid hormone biosynthesis, tryptophan metabolism, pentose and glucuronate interconversions, ascorbate and aldarate metabolism, as well as glutathione metabolism. Metabolic network diagram showed that the toxicity-related pathways were mainly associated with lysine metabolism in living organisms, glucuronic acid conversion, and hormone metabolism, especially the metabolism imbalance of lysine and glutathione would result in the disorder of energy metabolism or oxidative stress regulation, and thus inducing the damage in rats. Subacute toxicity test results for three doses groups (low, middle and high doses) showed that, Jinlingzi powder with doses of 19.7 g•kg⁻¹ and 39.4 g•kg⁻¹ caused obvious toxic effect, indicating Jinlingzi powder could produce toxic effect in vivo in a dose-dependent manner, and cause irreversible damage to the body.

Outcome Comparison of Nerve Transfer with Different Donor Nerves in a Rat Model.

OBJECTIVE: The phrenic nerve and the contralateral seventh cervical (C7) nerve root are the most commonly used donor nerves in the treatment of total brachial plexus avulsion. The aim of this study was to determine if the phrenic nerve or the contralateral C7 nerve root yields a superior outcome for nerve transfer. METHODS: A total of 60 Sprague-Dawley rats were randomly assigned to 1 of 3 groups. In Group A the phrenic nerve was used as the donor nerve; in Group B the contralateral C7 nerve root nerve was used as the donor nerve; in Group C the nerve was directly sutured. The results of behavioral assessment, electrophysiology, histology, nerve fiber count and muscle weight at 24 weeks postoperatively were recorded. RESULTS: Group A showed a faster recovery time compared to Group B; however Group B showed a better functional recovery at the final outcome assessment compared to Group A. CONCLUSION: The contralateral C7 nerve root was better as the donor nerve for nerve transfer in the treatment of total brachial plexus avulsion.

[Nuclear magnetic resonance-based metabonomics for sedative and hypnotic effect of Banxia Houpo decoction].

Insomnia was a common disease, which might be correlated with γ-aminobutyric acid A (GABAA) receptor mechanism, cytokine regulatory mechanism, excitatory amino acid mechanism and hydroxytryptamine (5-HT) receptor mechanism, but the correlations between these independent mechanisms and pathological characterization were still unclear. To further explore the effect of Banxia Houpo decoction on known or unknown biological pathways during treatment of insomnia, the metabonomics method based on ¹H-NMR was developed for detecting the significant changes in metabolomics after the administration with Banxia Houpo decoction in pentobarbital sodium-induced rat sleeping experiment. Serum and urine samples were analyzed by ¹H-NMR. Principal component analysis (PCA) was carried out for endogenous small molecule metabolites in urine and serum. H-NMR spectroscopies and relevant metabolites were found and identified by Simca-p 17.0 (Umet-rics, Umea, Sweden) and Chenomx NMR Suite 7.1 (Chenomx, Inc., Edmonton, Alberta, Canada) software. The result suggests that Banxia Houpo decoction group and indiplon group had significant differences. The load diagram showed the biggest variation metabolites and intergroup significant differences among 10 metabolic substances. According to the experiment, Banxia Houpo decoction group and indiplon group can prolonge the sleeping time of pentobarbital sodium-induced sprague-dawley rats, with a synergistic effect. The significant changes of these biomarkers indicated that the Banxia Houpo decoction could aid sleep by adjusting the content of glutamine, creatine phosphate, 2-oxoglutarate, and reducing the activity of brain nerves.

End-to-side neurotization with different donor nerves for treating brachial plexus injury: an experimental study in a rat model.

INTRODUCTION: End-to-side neurotization is currently used to treat brachial plexus injury, but it is not clear which donor nerve yields the best outcome. We performed experiments to determine the optimal donor nerve. METHODS: A total of 66 male Sprague-Dawley rats were assigned to 1 of 3 groups. Group A was the control group. In Group B, the phrenic nerve was used as the donor, while the ipsilateral C7 nerve root served as the donor in Group C. The epineurial window was used in end-to-side neurorrhaphy. Behavioral observations, histology, electrophysiology, and fluorescence retrotracing were performed postoperatively. RESULTS: Fluorescence retrotracing confirmed nerve regeneration in both Groups B and C upon end-to-side neurotization. The outcome of Group B was superior to that of Group C. CONCLUSIONS: Use of the phrenic nerve as the donor nerve yielded a better outcome than use of the ipsilateral C7 nerve root.

A Modified Endoscopic Carpal Tunnel Release Surgery.

In this study, a modified version of the endoscopic carpal tunnel release surgery was introduced, which is safe and easy to handle. Moreover, the requirement for surgical instruments is low. Six patients with carpal tunnel syndrome underwent the modified procedure. No neurovascular injuries occurred in these patients. According to the one-year follow-up data, all the patients were satisfied with the outcomes. The modified endoscopic carpal tunnel release technique has been proven to be safe with satisfactory outcomes in six patients in this study.

Local ectopic implantation for salvaging an amputated thumb: an anatomical study and a case report.

Relatively few cases of the ectopic implantation of an amputated thumb have been reported. The majority of ectopic sites are far from the original anatomic site, and anastomosis of the artery cannot be avoided in the secondary replantation surgery, which increases the failure rate. We conducted an anatomic study to investigate the possibility of performing local ectopic implantation of an amputated thumb at the wrist level such that an arterial pedicled thumb could be harvested in the secondary surgery. Thirty radial arteries were dissected from the point 3 cm proximal to the radial styloid to the point at which they passed over the radial styloid. Each branch with a diameter >0.8mm was selected. The mean number of branches was 3.2 > ± 1.37, and the mean diameter was 1.09 ± 0.83 mm. The mean diameter of the ulnar digital artery in the thumb was 1.08 ± 0.15 mm at the metacarpophalangeal joint level. Following anatomic research, an amputated thumb was ectopically implanted locally and replanted during a secondary operation without anastomosis of the artery. The amputated thumb survived, and its function was partially restored. In conclusion, the branches of the radial artery that have a similar diameter to the ulnar digital artery of the thumb can always be located at the wrist level. Local ectopic implantation is feasible and can help prevent the need for anastomosis of the artery during the secondary replantation surgery.

Preparation and characterization of a novel polymorph of indiplon, Form α.

A new polymorph α of indiplon was discovered, initially prepared by two methods, and further characterized by various means including single-crystal X-ray diffraction (SCXRD), powder X-ray diffraction (PXRD), variable temperature powder X-ray diffraction (VT-PXRD), differential scanning calorimetry (DSC), thermogravimetry analysis (TGA), Fourier transform Raman (FT-Raman) spectroscopy and solubility determination. The crystal structure of Form α as analyzed by SCXRD differ from the three previously reported polymorphs, Form I, II, and III. In addition, PXRD and solubility measurements could clearly distinguish between Form α and Form I. Slight differences between the two forms were also detected by FT-Raman. No differences between Form α and I were observed by DSC, which was explained by VT-PXRD results showing a solid-solid phase change from Form α to Form I during the heating process. Solubility measurements at various temperatures showed that the two polymorphs were mutually monotropic and that Form I was the relatively thermodynamically stable crystal form.

Analgesic Activity of Jin Ling Zi Powder and Its Single Herbs: A Serum Metabonomics Study.

OBJECTIVE: To compare the analgesic effect of Jin Ling Zi Powder (JLZ) and its two single herbs. METHODS: The hot plate method was used to induce pain. Totally 36 mice were randomly divided into 6 groups by a complete random design, including control, model, aspirin (ASP, 0.14 g/kg body weight), JLZ (14 g/kg body weight), Corydalis yanhusuo (YHS, 14 g/kg body weight), and Toosendan Fructus (TF, 14 g/kg body weight) groups, 6 mice in each group. The mice in the control and model groups were given the same volume of saline, daily for 2 consecutive weeks. At 30, 60, 90, and 120 min after the last administration, the pain threshold of mice in each group was measured, and the improvement rate of pain threshold was calculated. Serum endogenous metabolites were analyzed by gas chromatography-mass spectrometry (GC-MS). RESULTS: There was no statistical difference in pain threshold among groups before administration (P>0.05). After 2 weeks of administration, compared with the model group, the pain threshold in JLZ, YHS, TF and ASP groups were increased to varying degrees (P<0.05). JLZ had the best analgesic effect and was superior to YHS and TF groups. A total of 14 potential biomarkers were screened in serum data analysis and potential biomarkers levels were all reversed to different degrees after the treatment with JLZ and its single herbs. These potential biomarkers were mainly related to glyoxylate and dicarboxylate metabolism, glycine, serine and threonine metabolism, valine, leucine and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis and inositol phosphate metabolism. CONCLUSIONS: The analgesic mechanism of JLZ and YHS was mainly due to the combination of glycine and its receptor, producing post-synaptic potential, reducing the excitability of neurons, and weakening the afferent effect of painful information.

Determination and Difference Analysis of Phenolic Compounds in Smokers' Saliva and Mainstream Smoke.

To study the differences in phenolic compounds between tobacco smokers' saliva and mainstream smoke, a method was developed for the analysis of 12 phenolic compounds in saliva and mainstream smoke based on ultrahigh-performance liquid chromatography with fluorescence detection (UPLC-FLD). The contents and distributions of phenolic compounds in tobacco smokers' saliva and mainstream smoke were compared. The results were as follows: (1) Phenolic compounds were quantitatively analyzed by the internal standard method using 4-fluorophenol as an internal standard. For smokers' saliva samples, the limits of quantification (LOQs) ranged from 2.2 to 19.1 µg/L, and the recoveries were from 80.2% to 119.2% at the three spiked levels. For mainstream smoke samples, the LOQs ranged from 0.03 to 0.26 µg/cig, and the recoveries ranged from 84.9% to 107.0% at the three spiked levels. (2) The contents of phenolic compounds from 14 cigarettes in mainstream smoke and smokers' saliva were determined. In mainstream smoking, the main phenolic compounds were hydroquinone, catechol, phenol, meta- and para-Cresol, and o-methylhydroquinone. In smokers' saliva, the main phenolic compounds were phenol and meta- and para-Cresol and the contents of phenolic compounds in smokers' saliva from different cigarettes were significantly different. (3) The content distribution patterns of phenolic compounds in smokers' saliva differed from those in mainstream smoke. The predominant phenolic compound in mainstream smoke was dihydroxybenzene, while monophenols predominated in smokers' saliva. (4) The contents of phenolic compounds from five kinds of cigarettes were analyzed in the saliva of different smokers using principal component analysis, which indicated that cigarettes with different sensory effects were clearly distinguished by differences in the contents of phenolic compounds in saliva.

[A new triterpenoid fom Radix Pittospori].

To investigate chemical constituents from Radix Pittospori, chloroform extract of the roots was subjected to column chromatography with various chromatographic techniques. The structures were elucidated on the basis of physico-chemical property and spectral analysis. Two triterpenoids were identified as 22-acetyl-21-(2-acetoxy-2-methylbutanoyl)-R1-barrigenol(1) and 3alpha-hydroxyl-20-demethylisoaleuritolic-14(15)-ene-28, 30-dioic acid (2). Compound 1 is a new triterpene and compound 2 is isolated from this plant for the first time.

Transcription factor forkhead box K1 regulates miR-32 expression and enhances cell proliferation in colorectal cancer.

Increased microRNA (miR)-32 expression in colorectal cancer (CRC) tissues enhances CRC cell proliferation, migration, invasion and attenuates CRC cell apoptosis by repressing the expression of phosphatase and tensin homolog (PTEN). Forkhead box K1 (FOXK1) was identified as a potential interacting transcription factor using DNA pull-down assays and mass spectrometry. The present study aimed to elucidate the role of FOXK1 in regulating miR-32 expression in CRC. The expressions of FOXK1, miR-32, transmembrane protein 245 gene (TMEM245) and PTEN were compared between CRC and normal colonic tissues. Levels of miR-32, TMEM245, PTEN and the proliferation and apoptosis of CRC cells were studied using FOXK1-overexpression or knockdown, or by simultaneously interfering with FOXK1 and miR-32 expression. Direct FOXK1 binding to the miR-32 promoter was verified using chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. The results showed elevated FOXK1, miR-32 and TMEM245 expression, and significantly decreased PTEN expression in CRC, compared with normal colonic tissues. Correlations between the expressions of TMEM245 and miR-32, FOXK1 and miR-32, and FOXK1 and TMEM245 were positive and significant. FOXK1-knockdown led to decreased miR-32 and TMEM245 expression and increased PTEN expression, whereas FOXK1-overexpression had the opposite effect. Overexpressed FOXK1 promoted the malignancy of CRC cells in vitro by stimulating proliferation and reducing apoptosis; whereas FOXK1-depletion suppressed such malignancy and a miR-32 inhibitor partially reversed the effects of FOXK1. The results of ChIP and dual-luciferase reporter assays indicated that FOXK1 directly binds to the promoter of TMEM245/miR-32. Thus, the FOXK1-miR-32-PTEN signaling axis may play a crucial role in the pathogenesis and development of CRC.

Intercostal nerve transfer for restoration of the diaphragm muscle function after phrenic nerve transfer in total brachial plexus avulsion.

OBJECT: To determine the possibility of innervation of the diaphragm muscle using intercostal nerve after ipsilateral phrenic nerve transfer in total brachial plexus avulsion. METHODS: Bilateral phrenic nerves and the 9th intercostal nerves were observed inside the thorax. The point where the phrenic nerve entered the diaphragm muscle (point A), the point where the 9th intercostal nerve gave rise to the cutaneous branch (point B) and crossed the posterior axillary line (point C) and the point where the posterior axillary line met the insertion of the diaphragm muscle (point D) were identified. The distances between points B and C, points A and C and from points A through D to C were recorded respectively. The 9th intercostal nerve was transferred to the distal stump of the phrenic nerve in one patient after phrenic nerve transfer to avulsed brachial plexus. RESULTS: The mean distances between points B and C, points A and C and from points A through D to C were 12.20 ± 1.04 cm, 10.32 ± 1.02 cm and 16.43 ± 0.91 cm on the right side respectively, 11.78 ± 1.21 cm, 7.77 ± 0.85 cm and 11.74 ± 1.00 cm on the left side respectively. The 9th intercostal nerve was used to innervate the distal stump of the phrenic nerve in one patient after the phrenic nerve transfer to the avulsed brachial plexus. The diaphragm muscle function partially recovered one year after the operation. CONCLUSION: The 9th intercostal nerve can be transferred to the distal stump of the phrenic nerve to restore the diaphragm muscle function according to the anatomical study. The movement of the diaphragm muscle was partially restored in one clinical case.

Downregulation of microRNA-126 is inversely correlated with insulin receptor substrate-1 protein expression in colorectal cancer and is associated with advanced stages of disease.

Colorectal cancer (CRC) is a common human malignant tumor, and the fourth most common cause of cancer-associated mortality in China. However, the pathogenesis of CRC is not yet fully understood. The present study aimed to investigate the expression and clinical significance of microRNA (miR)-126 and insulin receptor substrate-1 (IRS-1), as well as the role of miR-126 in the prognosis of patients with CRC. A total of 86 colorectal tissue specimens, including 40 CRC and adjacent normal tissue, 26 colorectal adenoma tissue and 20 normal colorectal tissue samples, were collected for the present study. Reverse transcription-quantitative PCR analysis was performed to determine miR-126 and IRS-1 mRNA expression levels, while western blotting and immunohistochemistry (IHC) analyses were performed to determine IRS-1 protein expression levels. The correlation between miR-126 and IRS-1 expression, as well as the association between altered miR-126 and IRS-1 expression levels and clinicopathological characteristics, and the overall survival time of patients with CRC were assessed. The results demonstrated that miR-126 expression was significantly downregulated, while IRS-1 protein expression was upregulated in CRC tissues compared with that in adjacent normal tissues, colorectal adenoma tissues and normal colorectal tissues, respectively. IHC analysis exhibited strong positive staining of IRS-1 protein in CRC tissues, while absent or weak staining of IRS-1 protein was detected in adjacent normal tissues, colorectal adenoma tissues and normal colorectal tissues. miR-126 expression was inversely correlated with IRS-1 protein expression in CRC tissues (r=-0.420; P<0.05). Furthermore, downregulated miR-126 expression was associated with advanced clinicopathological characteristics of the disease and a shorter overall survival time in patients with CRC. Taken together, the results of the present study suggest that miR-126 downregulation may be a candidate molecular marker predictive of poor prognosis of patients with CRC.

The Effect of Potential on Surface Characteristic and Corrosion Resistance of Anodic Oxide Film Formed on Commercial Pure Titanium at the Potentiodynamic-Aging Mode.

Anodic oxidation treatment of commercially pure titanium was carried out at the voltages of 10, 30, 50 V in 0.5 M H2SO4 solution at the potentiodynamic-aging mode so as to obtain the effects of the anodic potential on the surface characteristic and corrosion resistance of the anodic oxide film. The influences of potential on the surface morphology, the roughness, the crystalline behavior, the chemical composition and the corrosion resistance of the anodic oxide films were investigated by using scanning electron microscopy (SEM), atomic force microscope (AFM), Raman spectrum, X-ray diffractometry (XRD), X-ray photoelectron spectroscopy (XPS), potentiodynamic polarization curves and electrode impedance spectroscopy (EIS). The results show that increasing anodic potential at the potentiodynamic-aging mode can significantly enhance thickness, flatness, crystallization, chemical stability, and corrosion resistance of anodic oxide film.

End-to-side neurotization with the phrenic nerve in restoring the function of toe extension: an experimental study in a rat model.

The phrenic nerve being transferred to the posterior division of the lower trunk with end-to-end neurorrhaphy is reported to be effective in restoring the function of digit extension in literature. However, the phrenic nerve is extremely important in respiration. We designed an animal experiment to discover whether the phrenic nerve being transferred to the posterior division of the lower trunk with end-to-side neurotization was feasible and provided the theoretical basis. A sum of 36 Sprague-Dawley rats was randomly assigned to one of two groups. In Group A, the phrenic nerve was transferred to the posterior division of the lower trunk with end-to-side neurotization. In Group B, the posterior division of the lower trunk was directly sutured. The results of behavioral assessment, electrophysiology, histology and nerve fiber count and muscle weight at 12 weeks postoperatively were recorded. In Group A, none of the rats experienced tachypnea. The motion of slight toe extension was observed. The results of electrophysiology, histology and nerve fiber count and muscle weight in Group A were not as well as those of Group B, but gradually improved with time. The phrenic nerve being transferred to the posterior division of lower trunk with end-to-side neurotization can partially restore the function of toe extension in a rat model. Whether the function of digit extension can be restored by the phrenic nerve with end-to-side neurotization in humans still needs more practice in clinic.