High levels of mitochondrial dynamics, autophagy, and apoptosis contribute to stable testicular status in hibernating Daurian ground squirrels.

Daurian ground squirrels (Spermophilus dauricus) experience various stress states during winter hibernation, but the impact on testicular function remains unclear. This study focused on the effects of changes in testicular autophagy, apoptosis, and mitochondrial homeostasis signaling pathways at various stages on the testes of Daurian ground squirrels. Results indicated that: (1) During winter hibernation, there was a significant increase in seminiferous tubule diameter and seminiferous epithelium thickness compared to summer. Spermatogonia number and testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels were higher during inter-bout arousal, suggesting that the testes remained stable during hibernation. (2) An increased number of mitochondria with intact morphology were observed during hibernation, indicating that mitochondrial homeostasis may contribute to testicular stability. (3) DNA fragmentation was evident in the testes during the hibernation and inter-bout arousal stages, with the highest level of caspase3 enzyme activity detected during inter-bout arousal, together with elevated levels of Bax/Bcl-2 and Lc3 II/Lc3 I, indicating an up-regulation of apoptosis and autophagy signaling pathways during hibernation. (4) The abundance of DRP1, MFF, OPA1, and MFN2 proteins was increased, suggesting an up-regulation of mitochondrial dynamics-related pathways. Overall, testicular autophagy, apoptosis, and mitochondrial homeostasis-related signaling pathways were notably active in the extreme winter environment. The well-maintained mitochondrial morphology may favor the production of reproductive hormones and support stable testicular morphology.

Idiomarina rhizosphaerae sp. nov. isolated from rhizosphere soil of Kalidium cuspidatum, and reclassification of Idiomarina andamanensis as Pseudidiomarina andamanensis comb. nov., and Idiomarina mangrovi as Pseudidiomarina mangrovi comb. nov.

A Gram-stain-negative, strictly aerobic, gliding motile, non-spore forming, rod-shaped indole-3-acetic acid producing bacterial strain, designated M1R2S28T, was isolated from the rhizosphere soil of Kalidium cuspidatum, in Tumd Right Banner, Inner Mongolia, China. Strain M1R2S28T grew at pH 5.0-10.0 (optimum 8.0), 10-45 °C (optimum 37 °C), in the presence of 0-20% (w/v) NaCl (optimum 5%). The phylogenetic trees based on the 16S rRNA gene sequences and the core-genome both revealed that strain M1R2S28T clustered tightly with Idiomarina loihiensis L2-TRT, and shared 99.3, 99.2, 98.7, and < 98.7% of the 16S rRNA gene sequence similarities with I. loihiensis L2-TRT, I. abyssalis KMM 227 T, I. ramblicola R22T, and all the other current type strains. The strain contained ubiquinone-8 (Q-8) as the sole respiratory quinone. Its major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminolipid, an unidentified phospholipid, and three unidentified lipids. Its major fatty acids were iso-C15:0, iso-C17:0, and iso-C17:1 ω9c. The genomic DNA G + C content was 46.8%. The average nucleotide identity based on BLAST (ANIb) and the digital DNA-DNA hybridization (dDDH) values of strain M1R2S28T to I. loihiensis L2-TRT, I. ramblicola R22T, and I. abyssalis KMM 227 T were 93.0, 82.9, and 81.8%, and 51.2, 26.0, and 25.0%, respectively. The phylogenetic and physiological results allowed the discrimination of strain M1R2S28T from its phylogenetic relatives. Idiomarina rhizosphaerae sp. nov. is, therefore, proposed with strain M1R2S28T (= CGMCC 1.19026 T = KCTC 92133 T) as the type strain. Additionally, based on the phylogenomic and genomic analysis results, Idiomarina andamanensis and Idiomarina mangrovi were transferred into genus Pseudidiomarina and be named Pseudidiomarina andamanensis comb. nov. and Pseudidiomarina mangrovi comb. nov., respectively.

Sinomicrobium kalidii sp. nov., an indole-3-acetic acid-producing endophyte from a shoot of halophyte Kalidium cuspidatum.

To better understand the effects of endophytic bacteria on halophytes, a bacteria that produced indole-3-acetic acid and 1-aminocyclopropane-1-carboxylic acid deaminase, designated HD2P242T, was isolated from a shoot of Kalidium cuspidatum collected in Tumd Right Banner, Inner Mongolia, PR China. The cells of strain HD2P242T were Gram-stain-negative, strictly aerobic, motile by gliding, non-spore-forming and rod-shaped. Strain HD2P242T grew at pH 6.0-9.0 (optimum, pH 7.0) and 10-45 °C (optimum 37 °C), in the presence of 0-8 % (w/v) NaCl (optimum, 4 %). The strain was positive for oxidase and catalase. The phylogenetic trees based on the 16S rRNA gene sequences and the whole genome sequences both showed that strain HD2P242T clustered with Sinomicrobium pectinilyticum 5DNS001T and S. oceani SCSIO 03483T, and had 95.6, 94.3 and <94.3 % 16S rRNA gene similarities to S. pectinilyticum 5DNS001T, S. oceani SCSIO 03483T and all the other current type strains. Strain HD2P242T contained menaquinone 6 as its sole respiratory quinone. Its major polar lipids were phosphatidylethanolamine, two unidentified aminolipids, two unidentified phospholipids and an unidentified lipid. The major fatty acids were iso-C17 : 0, iso-C16 : 0 3-OH, anteiso-C17 : 0 and summed feature 6 (C19 : 1 ω9c and/or C19 : 1 ω11c). The genome consisted of a 5 364 211 bp circular chromosome, with a G+C content of 45.1 mol%, predicting 4391 coding sequence genes, 47 tRNA genes and two rRNA operons. The average nucleotide identity based on blast and the digital DNA-DNA hybridization values of strain HD2P242T with S. oceani SCSIO 03483T and S. pectinilyticum 5DNS001T were 73.8 and 77.0%, and 22.3 and 22.2%, respectively. The comparative genome analysis showed that the pan-genomes of strain HD2P242T and three Sinomicrobium type strains possessed 4236 clusters, whereas the core genome possessed 2162 clusters, which accounted for 52.3 % of all the clusters. The genomic analysis revealed that all four Sinomicrobium members could utilize d-glucose by the glycolysis-gluconeogenesis pathway or the pentose phosphate pathway. The tricarboxylic acid cycle was utilized as a metabolic centre. The phylogenetic, physiological and phenotypic characteristics allowed the discrimination of strain HD2P242T from its phylogenetic relatives. Therefore, Sinomicrobium kalidii sp. nov. is proposed, and the type strain is HD2P242T (=CGMCC 1.19025T=KCTC 92136T).

Simultaneous determination of 16 important biologically active phytohormones in Dendrobium huoshanese by pressurized capillary electrochromatography.

A rapid pressurized capillary electrochromatography (pCEC) method has been successfully developed for the simultaneous determination of 16 phytohormones in Dendrobium huoshanense. Effects of wavelength, mobile phase, the flow rate, pH value, concentration of buffer and applied voltage were investigated, respectively. The results showed that the 16 phytohormones could be baseline-separated rapidly in less than 21 min on a reversed-phase EP-100-20/45-3-C18 capillary column (total length of 45 cm, effective length of 20 cm, diameter of 100 µm, ODS packing inside for 3 µm) with ACN/5.0 mM ammonium acetate (containing 0.05% formic acid, pH = 3) as the mobile phase using gradient elution mode as follows: 0.1-10.0 min 40%ACN,10-15.0 min 70%ACN, 15.0-20 min 80% ACN, 20-21.0 min 80% ACN at a flow rate of 0.12 mL/min, applied voltage of -5 kV and a UV detection wavelength of 210 nm. The method validation howed that the established method is precise and stability, and the RSDs of intra- and inter-day precision based retention time and peak area were all below 5%. Employed the established method, in our experimental conditions, total 6 endogenous hormones including IAA, IBA, NAA, GA, ABA, t-Z were detected in D. huoshense. However, a relative larger amount of exogenous hormone 2,4-D (25.3 ~ 4.2 µg/kg) and 6-BA (79.5 ~ 35.4 µg/kg) were detected in 1 ~ 4 year old cultivated D. huoshense, suggesting there were still a certain amount of exogenous hormone residue in tissue-cultured D. huoshanese though they had been transplanted to field cultivation from the test-tube plantlets for several years.

Neuroprotective effects of madecassoside in early stage of Parkinson's disease induced by MPTP in rats.

In this study, we investigated the neuroprotective effects of madecassoside, isolated from the Chinese medicinal herb Centella asiatica, in the rat model of early phase of parkinsonism. During intragastric administrations of madecassoside for 7 days, the rats were injected with MPTP on the 7th day. And for the following 14 days, madecassoside were also administered. On the 14th day, the behavioral tests were assessed after 1h of administration. And then, the rats were sacrificed, substantia nigra and striatum were dissected. The content of DA, MDA, GSH, and Bcl-2/Bax gene expression levels and BDNF protein level was determined. Treatment with madecassoside was found to improve locomotor dysfunction and to protect dopaminergic neuron by antagonizing MPTP induced neurotoxicity. Madecassoside significantly attenuated the MPTP-induced reduction of dopamine in the striatum. The MDA contents were significantly decreased while the GSH levels, Bcl-2/Bax ratio and protein expression of BDNF were significantly increased in madecassoside treated groups. These results indicated that madecassoside was effective in recovering MPTP-induced early signs of parkinsonism via its neuroprotective effects including reversing the depletion of DA, antioxidant activity, increasing ratio of Bcl-2/Bax, increasing protein expression of BDNF.

Magnolol treatment reversed the glial pathology in an unpredictable chronic mild stress-induced rat model of depression.

Growing evidence indicates that glia atrophy contributes to the pathophysiology and the pathogenesis of major depressive disorder. Magnolol is the main constituent identified in the bark of Magnolia officinalis, which has been used for the treatment of mental disorders, including depression, in Asian countries. In this study, we investigated the antidepressant-like effect and the possible mechanisms of magnolol in rats subjected to unpredictable chronic mild stress (UCMS). The ameliorative effect of magnolol on depression symptoms was investigated through behavior tests, including sucrose preference test, open-field test and forced-swimming test. In addition, the levels of glial fibrillary acidic protein (GFAP), an astrocyte marker, in the hippocampus and prefrontal cortex were determined by immunohistochemistry, Western blot, and reverse transcription-polymerase chain reaction (RT-PCR). Exposure to UCMS resulted in a decrease of behavioral activity, whereas magnolol (20, 40 mg/kg) and fluoxetine (20mg/kg) administration significantly reversed the depressive-like behaviors (P<0.05).Moreover, treatment with magnolol effectively increased GFAP mRNA and protein levels in UCMS rats. These results confirmed the antidepressant-like effect of magnolol, which maybe primarily mediated by reversing the glial atrophy in the UCMS rat brain.

Antidepressant-like effect of magnolol on BDNF up-regulation and serotonergic system activity in unpredictable chronic mild stress treated rats.

Magnolol is the main constituent identified in the barks of Magnolia officinalis, which has been used for the treatment of mental disorders including depression in China. In this study, we investigated the antidepressant-like effect of magnolol, and its possible mechanisms in rats subjected to unpredictable chronic mild stress (UCMS). High performance liquid chromatography with electrochemical detection (HPLC-ECD) and immunohistochemical staining analysis were applied to explore the mechanisms underlying the antidepressant-like effect of magnolol. Magnolol (20, 40 mg/kg) significantly reversed UCMS-induced reduction in sucrose consumption and deficiency in locomotor activity. In addition, it was observed that administration of magnolol (20, 40 mg/kg) restored brain-derived neurotrophic factor (BDNF) expression, and normalized the serotonergic system changes in the UCMS-treated rats. These results confirmed the antidepressant-like effect of magnolol, which might be based primarily on its ability to increase the BDNF expression and enhance the activity of the serotonergic system in rat brains.

Asiaticoside: attenuation of neurotoxicity induced by MPTP in a rat model of Parkinsonism via maintaining redox balance and up-regulating the ratio of Bcl-2/Bax.

In this study, we investigated the neuroprotective effects of asiaticoside, a triterpenoid saponin isolated from the Chinese medicinal herb Centella asiatica, in the rats model of Parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Rats were first injected with MPTP. One day after surgery, asiaticoside was administered and the behavioral tests were assessed. On 14th day, the rats were sacrificed, substantia nigra (SN) and striatum were dissected, and then dopamine (DA) and its metabolites in striatum and malonyldialdehyde (MDA) contents, reduced glutathione (GSH) level and gene expression level in SN were estimated. Treatment with asiaticoside was found to protect dopaminergic neuron by antagonizing MPTP induced neurotoxicity and to improve locomotor dysfunction. Asiaticoside significantly attenuated the MPTP-induced reduction of dopamine in the striatum. The content of MDA was significantly decreased while the GSH level was significantly increased in asiaticoside-treated groups. In addition, asiaticoside increased the Bcl-2/Bax ratio. These results indicated that asiaticoside was effective in reversing MPTP induced Parkinsonism via its neuroprotective effects including antioxidant activity, maintaining the metabolic balance of DA, and increasing ratio of Bcl-2/Bax.

Antidepressant-like effects of the saponins extracted from Chaihu-jia-longgu-muli-tang in a rat unpredictable chronic mild stress model.

Chaihu-jia-longgu-muli-tang (CLM) has been used for treating depressive disorders for thousands of years in China. In the present study, we investigated the antidepressant-like effect of the saponins extracted from CLM (SCLM) in rats subjected to unpredictable chronic mild stress (UCMS). The ameliorative effect of SCLM on symptom of depression through behavior tests including: sucrose preference test, open-field test and forced-swimming test was investigated. In addition, high performance liquid chromatography with electrochemical detection (HPLC-ECD), immunohistochemical staining analysis and RT-PCR were applied to explore the mechanisms underlying the antidepressant-like effects of SCLM. It was observed that administration of SCLM (70, 140 mg/kg) reversed the depressive-like behaviors, restored the reduction in the levels of monoamine neurotransmitters and up-regulated the expression of brain-derived neurotrophic factor (BDNF) in UCMS-treated rats. These findings confirmed the antidepressant-like effects of SCLM in UCMS model of rats.

Protective effect of Zhen-Wu-Tang (ZWT) through keeping DA stable and VMAT 2/DAT mRNA in balance in rats with striatal lesions induced by MPTP.

ETHNOPHARMACOLOGICAL RELEVANCE: Zhen-Wu-Tang (ZWT), the modified formulation of a classical Chinese prescription from "Treaties on Febrile Disease", was clinically employed to treat Parkinson's disease. AIM OF THE STUDY: To investigate the neuroprotective effect of ZWT on intra-striatum injection of MPTP-induced Parkinson's disease in rats. MATERIALS AND METHODS: The effect of ZWT on the behavioral changes (open-field test, Ladder walking, spontaneous alternation in Y maze), the dopamine transmitter systems of substantia nigra, striatum and frontal cortex of rats by HPLC-ECD, mRNA expression of tyrosine hydroxylase (TH), dopamine transporter (DAT) and vesicular monoamine transporter 2 (VMAT 2) of the above three brain regions was investigated. RESULTS: This study showed that ZWT not only ameliorated the behavior induced by the administration of MPTP in striatum, but also increased DA in the brain, prevented the decreasing of TH and balanced the ratio of VMAT 2/DAT in mRNA level. CONCLUSIONS: These results suggest that ZWT possesses neuroprotective and anti-parkinsonism properties.

Ameliorative and neuroprotective effect in MPTP model of Parkinson's disease by Zhen-Wu-Tang (ZWT), a traditional Chinese medicine.

AIM OF THE STUDY: Traditional Chinese medicine Zhen-Wu-Tang (ZWT) is a well-known PentaHerbs formula from "Treatise on Febrile Disease". This study is to elucidate its neuroprotective effect and mechanism of ameliorative effect of the syndrome of Parkinson's disease (PD). MATERIALS AND METHODS: The ameliorative effect of ZWT on symptom of PD through behavior tests including: swimming test, the tail suspension test and open-field test was investigated. The neuroprotective effect of dopaminergic neurons from the striatum and frontal cortex of brain was detected by high performance liquid chromatography with electrochemical detection (HPLC-ECD). RESULTS: This study proved that ZWT could ameliorate the typical symptom of PD and protect dopaminergic system. CONCLUSION: These results suggested that ZWT possessed protective and ameliorative properties of dopaminergic neurons.