[Effects of serum containing Shenrong Pill on oxidative stress damage in mouse Leydig cells].

OBJECTIVE: To explore the mechanism of Shenrong pills in improving oligoasthenospermia by inhibiting oxidative stress-induced Leydig cell apoptosis in mouse testis. METHODS: The oxidative stress model of mouse Leydig cells (TM3) was induced by 3% hydrogen peroxide (H2O2) of 600 µmol/L, and then TM3 cells were treated with 7.5%, 10% and 12.5% serum containing Shenrong pills, respectively. TM3 cells were divided into normal control group, model group, and low, medium and high dose groups of Shenrong pills containing serum. The cell viability of TM3 after treatment was detected by CCK-8 method, reactive oxygen species (ROS) were detected by DCFH probe method, and SOD-1, CAT, GSH-px, MDA and LPO in cell lysates were detected by ELISA method. The changes of apoptosis-related proteins Bcl-2, Bax in cell lysates were detected by Western Blot. RESULTS: After H2O2 treatment, compared with normal control group, cell viability was significantly decreased (P< 0.01), MDA and LPO contents were significantly increased (P<0.01), SOD-1, CAT and GSH-px contents were significantly decreased (P<0.01). Reactive oxygen species (ROS) were significantly increased, the relative expression ratio of Bcl-2 protein was significantly decreased (P<0.01, P<0.05), and the relative expression of Bax protein was increased. After the administration of Shenrong pills containing serum, the above indexes were reversed to varying degrees. CONCLUSION: Shenrong pills can resist oxidative stress, inhibit the apoptosis of TM3 cells in mice, maintain high levels of testosterone required for spermatogenic cells, and improve the sperm quality of mice with oligonasthenospermia.

[Medication experience of TCM Master Academician Wang Qi in the treatment of premature ejaculation: Data mining and analysis].

Objective: To analyze the academic thought, medication experience and prescription rules of Academician Wang Qi in the treatment of premature ejaculation (PE) using the TCM inheritance support platform (V2.5). METHODS: We collected and sorted out the medical records on the treatment of PE from Academician Wang Qi's Clinic. We established a database of medical records on the TCM inheritance support platform, analyzed the drugs and prescriptions in the database and explored new prescriptions using "statistical reports" and "data analysis" systems on the platform. RESULTS: A total of 91 effective prescriptions were recorded, involving 148 TCM drugs, with Phellodendron, Amomum Villosum, Polygala Tenuifolia, Tuckahoe, Lodestone, Oyster, Acanthopanax Senticosus, Uncaria, Tribulus, and Keel as the top 10 with the highest frequency of use, which were featured mainly by "warm" and "cold" concerning the four natures, "sweet", "bitter" and "pungent" relating to the five flavors, and acting on "kidney meridian", "liver meridian" and "heart meridian" in terms of the meridian tropisms. In addition, 5 new prescriptions were obtained through unsupervised entropy hierarchical clustering. CONCLUSIONS: In the treatment of PE, Academician Wang Qi employs tranquilizing the mind and consolidating the kidney (An Zhi Gu Shen) as the primary strategy, taking into account the three organs of heart, liver and kidneys, focusing on the phase of calming the mind or regulating the liver or clearing the kidney or controlling fire, and adding or reducing drugs according to different conditions and syndromes, which conforms to his diagnosis and treatment mode of "body differentiation－disease differentiation－syndrome differentiation". The analysis of the potential new prescriptions also accords with Academician Wang Qi's rules of medication, which can provide some ideas for the clinical treatment of and scientific researches on premature ejaculation in the future.

[Academician Wang Qi's medication rules for oligoasthenozoospermia: An analysis based on the Traditional Chinese Medicine Inheritance Auxiliary Platform].

OBJECTIVE: To analyze the medication rules for oligoasthenozoospermia (OAZ) observed by Wang Qi, an academician, master of Traditional Chinese Medicine (TCM) and initiator of andrology in TCM. METHODS: We collected the outpatient cases of OAZ treated by Wang Qi and established a database of clinical medical records using the TCM Inheritance Auxiliary Platform. Employing the integrated rule-based system for analysis of the software, we modified the mutual information method, complex system entropy clustering analysis and other data mining methods, and summarized the medication rules Wang Qi followed in the treatment of OAZ. RESULTS: A total of 134 prescriptions made by Wang Qi for the treatment of OAZ were collected, involving 110 TCM drugs, which are mainly neutral and warm in nature and taste sweet and mostly act through the liver and kidney meridians. The core formula ingredients of the prescriptions included Morinda officinalis, Cuscuta chinensis, Lycium barbarum, Mulberry, Angelica sinensis, Astragalus mongholicus and Fish Maw, and most frequently Morinda officinalis, Cuscuta chinensis, Lycium barbarum and Mulberry. CONCLUSIONS: Wang Qi holds that kidney deficiency, dampness-heat, blood stasis and toxin are the main pathogenic factors for OAZ. The basic treatment of OAZ is to invigorate the kidney and replenish the essence, and meanwhile activate blood circulation, dissipate stasis and eliminate dampness-heat.

Synthesis and evaluation of 1,2,3,4-tetrahydro-1-acridone analogues as potential dual inhibitors for amyloid-beta and tau aggregation.

Amyloid-ß (Aß) and tau protein are two crucial hallmarks in Alzheimer's disease (AD). Their aggregation forms are thought to be toxic to the neurons in the brain. A series of new 1,2,3,4-tetrahydro-1-acridone analogues were designed, synthesized, and evaluated as potential dual inhibitors for Aß and tau aggregation. In vitro studies showed that compounds 25-30 (20 µM) with N-methylation of the quinolone ring effectively inhibited Aß1-42 aggregation by 84.7%-99.5% and tau aggregation by 71.2%-101.8%. Their structure-activity relationships are discussed. In particular, 30 could permeate the blood-brain barrier, bind to Aß1-42 and tau, inhibit Aß1-42 ß-sheets formation, and prevent tau aggregation in living cells.

[Bazi Granules improves sperm quality in rats with oligoasthenozoospermia induced by multi-glycosides of tripterygium wilfordii].

OBJECTIVE: To investigate the effect of Bazi Granules on sperm quality in male rats with oligoasthenozoospermia (OAS) induced by multi-glycosides of tripterygium wilfordii (GTW). METHODS: Thirty-six SD male rats were randomly divided into six groups of equal number: normal control, OAS model control, Wuziyanzong Pills (WYP), and low-, medium- and high-dose Bazi. The OAS model was established in the rats except those of the normal control group by intragastrical delivery of GTW at 30 mg/kg/d for 40 days. From the 41st day, the animals of the normal and OAS model control groups were fed with distilled water, those of the WYP group treated by gavage with WYP at 1.02 g/kg/d, and those of the low-, medium- and high-dose Bazi groups intragastically given Bazi Granules 3 (5.27 g/kg), 6 (10.54 g/kg) and 12 (21.08 g/kg) times, respectively, that of the human-equivalent dose. Semen parameters and the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in the testis tissue were determined after 28 days of treatment. RESULTS: After treatment, the rats of the of the high-, medium- and low-dose Bazi groups, compared with the OAS model controls, showed significant increases in sperm concentration (ï¼»1 050.71 ± 203.71ï¼½, ï¼»1 370.06 ± 166.01ï¼½ and ï¼»1 302.53 ± 476.51ï¼½ vs ï¼»617.01 ± 237.08ï¼½ ×106/ml, P < 0.05), sperm motility (ï¼»0.56 ± 0.24ï¼½%, ï¼»0.73 ± 0.14ï¼½% and ï¼»0.70 ± 0.23ï¼½% vs ï¼»0.07 ± 0.05ï¼½%, P < 0.05), sperm average path velocity (ï¼»85.71 ± 30.35ï¼½, ï¼»83.83 ± 10.31ï¼½ and ï¼»75.06 ± 19.70ï¼½ vs ï¼»43.45 ± 38.74ï¼½ µm/s, P < 0.05), sperm curvilinear velocity (ï¼»101.76 ± 23.28ï¼½, ï¼»119.60 ± 21.22ï¼½ and ï¼»102.11 ± 32.89ï¼½ vs ï¼»53.63 ± 47.91ï¼½ µm/s, P < 0.05), sperm straight line velocity (ï¼»62.75 ± 7.63ï¼½, ï¼»67.80 ± 5.05ï¼½ and ï¼»64.11 ± 12.03ï¼½ vs ï¼»40.18 ± 36.86ï¼½ µm/s, P < 0.05), and the SOD level (ï¼»380.23 ± 75.07ï¼½, ï¼»349.53 ± 97.48ï¼½ and ï¼»415.07 ± 72.01ï¼½ vs ï¼»304.62 ± 27.17ï¼½ U/mg, P < 0.05), but a remarkable decrease in the MDA level (ï¼»0.33 ± 0.16ï¼½, ï¼»0.22 ± 0.05ï¼½ and ï¼»0.34 ± 0.22ï¼½ vs ï¼»0.73 ± 0.20ï¼½ nmol/mg, P < 0.05). CONCLUSIONS: Bazi Granules can significantly improve the sperm quality of OAS rats, which may be related to its abilities of repairing oxidative stress injury and enhancing oxidation resistance.

[Protective effect of jiedu tongluo injection on cerebral edema in rats with lesion of cerebral ischemia/reperfusion].

OBJECTIVE: To investigate the protective effects of Jiedu Tongluo injection on cerebral edema induced by focal lesion of cerebral ischemia/reperfusion, the hydrous content of brain and the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin and MMP-9 in rats. METHOD: The model of brain middle cerebral artery ischemia/reperfusion was established by the thread approach. After 24 hours of reperfusion, cerebral edema formation was determined by the hydrous content of brain. The permeability of blood brain barrier was evaluated based on the leakage of Evans blue. Enzyme-linked immunoadsordent assay (ELISA)was used to examine the expression of ICAM-1, VCAM-1, E-selectin. The expression of MMP-9 was measured by immunohistochemistry. RESULT: JDTL, in the dose of 2 mL x kg(-1) and 4 mL x kg(-1), relieved cerebral edema (P < 0.05, P < 0.01), reduced the expressions of ICAM-1, VCAM-land E-selectin and decreased MMP-9 activity (P < 0. 05, P < 0.01) in model rats. CONCLUSION: Jiedu Tongluo injection has a protective effect on rat brain from cerebral edema induced by the injury of focal cerebral ischemia/reperfusion. The mechanism is related to that Jiedu Tongluo injection can reduce the expressions of ICAM-1, VCAM-1 and E-selectin and inhibit of MMP-9 activation in rat brain.

Anti-inflammation and antioxidant effect of Cordymin, a peptide purified from the medicinal mushroom Cordyceps sinensis, in middle cerebral artery occlusion-induced focal cerebral ischemia in rats.

Cordymin is a peptide purified from the medicinal mushroom Cordyceps sinensis. The present study investigated the effects of Cordymin in prevention of focal cerebral ischemic/reperfusion (IR) injury. The right middle cerebral artery occlusion model was used in the study. The effects of Cordymin on mortality rate, neurobehavior, grip strength, glutathione content, lipid Peroxidation, glutathione peroxidase activity, glutathione reductase activity, catalase activity, Na(+)K(+)ATPase activity glutathione S transferase activity and on the regulation of C3 and C4 protein level, polymorphonuclear cells, interleukin-1ß and tumor necrosis factor-α in a rat model were studied respectively. Treatment (orally) of Cordymin significantly boosted the defense mechanism against cerebral ischemia by increasing antioxidants activity related to lesion pathogenesis. Restoration of the antioxidant homeostasis in the brain after reperfusion may have helped the brain recover from ischemic injury. Moreover, Cordymin significantly inhibited infiltration of polymorphonuclear cells and IR-induced up-regulation of the brain production of C3 protein level, interleukin-1ß and tumor necrosis factor-α. Cordymin significantly improved the outcome in rats after cerebral ischemia and reperfusion in terms of neurobehavioral function. Our findings suggest that cordymin have a neuroprotective effect in the ischemic brain, which is due to the inhibition of inflammation and increase of antioxidants activity related to lesion pathogenesis. Cordymin can be used as potential preventive agent against cerebral ischemia-reperfusion injury.

Isolation and biological activity of triglycerides of the fermented mushroom of Coprinus Comatus.

BACKGROUND: Although many physiological functions of Coprinus comatus have been reported, there has been no report on the antinociceptive activity of Coprinus comatus. Therefore, the objective of the present study is to demonstrate the production, isolation, and biological properties of triglycerides (TFC) of the fermented mushroom of Coprinus comatus. METHODS: The effects of TFC on cytokines levels, total antioxidant activity, antinociceptive effects in vivo, LD50 and tactile hyperalgesia were analyzed respectively. RESULTS: TFC treatment decreased the levels of cytokines and total antioxidant status (TAOS) and inhibited the acetic acid-induced abdominal constrictions in mice. In addition, TFC reduced CFA-induced tactile hyperalgesia in a dose-dependent manner and the LD50 of TFC was determined to be 400 mg/kg. However, TFC did not significantly inhibit the reaction time to thermal stimuli in the hot-plate test. CONCLUSIONS: TFC showed anti-inflammatory, antioxidant, peripheral antinociceptive and antihyperalgesic activity in various models of inflammatory pain. The data suggest that TFC may be a viable treatment option for inflammatory pain.

[Preparation and SERS activity studies on substrate of nano-silver film with broad plasma absorption band].

A highly reproducible and broad plasma absorption band nano-silver film was prepared by electrostatic self-assembly technology. The size of nano-silver particles on the surface of silver film, in the scanning electron microscopy (SEM) micrograph, is in a wide distribution from 18 to 200 nm. At the same time, the ultraviolet-visible spectrophotometer was employed to detect the nano-silver film. There is a broad plasma absorption band in the UV-Vis spectrum. To test the SERS activity of the nano-silver film, crystal violet and serum of healthy person were used as probe molecules and the high quality SERS spectra were obtained.

The protective effect of cordyceps sinensis extract on cerebral ischemic injury via modulating the mitochondrial respiratory chain and inhibiting the mitochondrial apoptotic pathway.

Cerebral ischemia is a common refractory brain disease, resulting from a reduction in the blood flow to the brain. Mitochondrial dysfunction leads to ischemic stroke and brain injury. Cordyceps sinensis (CS) is an important traditional Chinese medicine, which has been linked to neuroprotection in recent studies. In this study, we investigated the role of the mitochondrial respiratory chain and the mitochondrial apoptotic pathway on the protective effect of Cordyceps sinensis extract (CSE) against cerebral ischemia injury both in vivo and in vitro. In a murine middle cerebral artery occlusion (MCAO) model, administration of CSE relieved neuronal morphological damage and attenuated the neuronal apoptosis. CSE also reduced neurobehavioral scores and oxygen free radical (OFR), while improving the levels of ATP, cytochrome c oxidase (COX), and mitochondrial complexes I-IV. Furthermore, the mRNA expression of Bax, cytochrome c (Cyt c) and caspase-3 were down-regulated. In brain microvascular endothelial cells (BMECs) exposed to oxygen and glucose deprivation (OGD), CSE prevented OGD-induced cellular apoptosis, and recovered the reduction of mitochondrial membrane potential (MMP). Moreover, CSE treatment induced an increase of Bcl-2 protein expression and a decrease of Bax, Cyt c and caspase-3 protein expression. Meanwhile, the caspase-3, -8, and -9 activities were also inhibited. The results indicate that CSE can relieve cerebral ischemia injury and exhibit protective effects via modulating the mitochondrial respiratory chain and inhibiting the mitochondrial apoptotic pathway.

Design, synthesis and evaluation of 2-arylethenyl-N-methylquinolinium derivatives as effective multifunctional agents for Alzheimer's disease treatment.

A series of 2-arylethenyl-N-methylquinolinium derivatives were designed and synthesized based on our previous research of 2-arylethenylquinoline analogues as multifunctional agents for the treatment of Alzheimer's disease (AD) (Eur. J. Med. Chem. 2015, 89, 349-361). The results of in vitro biological activity evaluation, including ß-amyloid (Aß) aggregation inhibition, cholinesterase inhibition, and antioxidant activity, showed that introduction of N-methyl in quinoline ring significantly improved the anti-AD potential of compounds. The optimal compound, compound a12, dramatically attenuated the cell death of glutamate-induced HT22 cells by preventing the generation of ROS and increasing the level of GSH. Most importantly, intragastric administration of a12•HAc was well tolerated at doses up to 2000 mg/kg and could traverse blood-brain barrier.

Discovery of Small Molecules for Up-Regulating the Translation of Antiamyloidogenic Secretase, a Disintegrin and Metalloproteinase 10 (ADAM10), by Binding to the G-Quadruplex-Forming Sequence in the 5' Untranslated Region (UTR) of Its mRNA.

Up-regulation of a disintegrin and metalloproteinase 10 (ADAM10) to prevent the formation of ß-amyloid (Aß) peptides might be a promising strategy to treat Alzheimer's disease (AD). RNA G-quadruplex motif within the 5'-UTR of the ADAM10 mRNA is an inhibitory element for ADAM10 translation. Thus, mitigation of the suppressive effect of this motif using an RNA G-quadruplex-forming G-rich sequence (QGRS) binder might be a new approach for AD therapy. Herein, a series of new methylquinolinium derivatives were synthesized and screened by surface plasmon resonance (SPR) and the dual-luciferase reporter assay. Among them, compound 24 showed selective affinity for the QGRS of ADAM10 and could strongly up-regulate the translation of it. Moreover, treatment with 24 led to a significant increase of the secretion of sAPPα, consequently decreasing the Aß40 in cellular. These results illustrate that the interaction between the RNA QGRS and a small molecule may be a new molecular strategy to modulate the translation of ADAM10.