Response of soil stoichiometric characteristics to climatic factors in temperate steppe of Longzhong region, China.

We explored the relationship between climate factors (mean annual precipitation and mean annual temperature) and the contents and stoichiometry of soil carbon (C), nitrogen (N), and phosphorus (P) at different soil depths (0-5, 5-10, 10-20, 20-30, 30-50, 50-70, and 70-100 cm) temperate steppe of Longzhong. The results showed with the increases of soil depth, soil C, N contents, C:P, and N:P gradually decreased from 21.88 g·kg-1, 1.84 g·kg-1, 33.6 and 3.1 to 7.67 g·kg-1, 0.59 g·kg-1, 12.5 and 1.0, respectively. Soil C:N showed an increasing trend from 12.2 to 13.9, while soil P content remained stable with an average of 0.61 g·kg-1. Soil C, N, C:P, and N:P were significantly positively correlated with mean annual precipitation and negatively correlated with mean annual temperature. Soil P content and C:N were not correlated with mean annual precipita-tion and mean annual temperature. With the increases of soil depth, the total explanatory power of the changes in soil C, N and P contents by mean annual precipitation and mean annual temperature decreased and then increased, and that in soil C:P, N:P and C:N did not change significantly. The changes of soil C, N and P contents on the temperature steppe were mainly influenced by mean annual precipitation. The effects and relative contributions of mean annual precipitation and mean annual temperature on the variations of soil nutrient contents and stoichiometry of C, N and P differed at different soil depths.

The impact of lactating Hu sheep's dietary protein levels on lactation performance, progeny growth and rumen development.

This study aimed to investigate whether lactating Hu sheep's dietary protein levels could generate dynamic effects on the performance of their offspring. Twelve ewes with similar parity were fed iso-energy diets which contained different protein levels (P1: 9.82%, P2: 10.99%) (n = 6), and the corresponding offspring were divided into SP1 and SP2 (n = 12). At 60 days, half of the lambs were harvested for further study: the carcass weight (p = 0.043) and dressing percentage (p = 0.004) in the SP2 group were significantly higher than SP1. The acetic acid (p = 0.007), propionic acid (p = 0.003), butyric acid (p < 0.001) and volatile fatty acids (p < 0.001) in rumen fluid of SP2 were significantly lower than SP1. The expression of MCT2 (p = 0.024), ACSS1 (p = 0.039) and NHE3 (p = 0.006) in the rumen of SP2 was lower than SP1, while the HMGCS1 (p = 0.026), HMGCR (p = 0.024) and Na+/K+-ATPase (p = 0.020) was higher than SP1. The three dominant phyla in the rumen are Bacteroidetes, Proteobacteria and Firmicutes. The membrane transport, amino acid metabolism and carbohydrate metabolism of SP1 were relatively enhanced, the replication and repair function of SP2 was relatively enhanced. To sum up, the increase of dietary protein level significantly increased the carcass weight and dressing percentage of offspring and had significant effects on rumen volatile fatty acids, acetic acid activation and cholesterol synthesis related genes. HIGHLIGHTSIn the early feeding period, the difference in ADG of lambs was mainly caused by the sucking effect.The increase in dietary protein level of ewes significantly increased the carcass weight and dressing percentage of offspring.The dietary protein level of ewes significantly affected the volatile fatty acids (VFAs) and genes related to acetic acid activation and cholesterol synthesis in the rumen of their offspring.The membrane transport, amino acid metabolism and carbohydrate metabolism of the offspring of ewes fed with a low protein diet were relatively enhanced.The replication and repair function of the offspring of ewes fed with a high protein diet was relatively strengthened.

Aroma effects of key volatile compounds in Keemun black tea at different grades: HS-SPME-GC-MS, sensory evaluation, and chemometrics.

The present study aimed to explore the relationship between the grade and the characteristic aroma in Keemun black tea (KBT). Headspace solid-phase microextraction (HS-SPME), gas chromatography-mass spectrometry (GC-MS), sensory evaluation, and chemometrics were employed to determine the changes in the flavor evolution of KBT at grade. The results showed that a total of 110 volatile components were identified. Linalool and linalool oxide were dominant. The orthogonal partial least squares discriminant analysis (OPLS-DA) combined with relative odor activity value (rOAV > 0.1) revealed that 11 volatile components were the key volatile compounds of KBT, such as benzeneacetaldehyde (rOAV: 3.43-5.96) and methyl salicylate (rOAV: 2.15 - 2.50). Furthermore, the partial least squares (PLS) model indicated that geraniol, linalool, and methyl salicylate benefited from the reservation of floral flavor of Keemun aroma characteristic of KBT. The findings presented in this thesis add to our understanding of KBT at different grades.

Isolation, identification, and quantification of triterpene saponins in the fresh fruits of Panax notoginseng.

This study is to develop a method for isolation, identification, and quantitative determination of dammarane-type triterpene saponins in the Panax notoginseng fruits (PNF). The saponins were isolated by a serious of chromatographic methods, and their structures were elucidated on the basis of spectroscopic evidence and comparison with those of literature reports. Quantitative assay was performed on an ultra-performance liquid chromatography-UV (UPLC-UV) method. As a result, 22 saponins were isolated from the extract of PNF, among them, compound 1 was a new saponin, named as malonylgypenoside IX, compounds 3-10, and 14-18 were isolated from the PNF for the first time. As to quantitative analysis, the calibration curves showed good linearity (r > 0.998) within the concentration range, and the method validation provided good reproducibility and sensitivity for the quantification of eight major saponins with precision and accuracy of less than 3.0%.

New malonylginsenosides from the fresh fruits of Panax notoginseng.

Four new malonylginsenosides, malonylnotoginsenoside Fe (1), malonylnotoginsenoside Ra1 (2), malonylgypenoside LXXV (3), and malonylginsenoside Mc (4), together with two known analogues, malonylfloralginsenoside Rc1 (5) and malonylginsenoside Rc (6), were isolated from the fresh fruits of Panax notoginseng. Their structures were determined by MS and NMR experiments. The anti-proliferative activities of the malonylginsenosides (1-6) against SH-SY5Y human neuroblastoma cell line were evaluated using the MTT assay.

Clinical Observation of Effects of Modified Xiangsha Liujunzi Tang Application on Adverse Reaction of Moderately/Highly Emetogenic Chemotherapy for Gastrointestinal Tumors / 中国实验方剂学杂志

Objective:To investigate the effects of external application of modified Xiangsha Liujunzi Tang in improving gastrointestinal reaction，immune function， and quality of life in patients with gastrointestinal tumors after moderately/highly emetogenic chemotherapy. Method:A total of 140 inpatients （from January 2018 to May 2021） with gastrointestinal malignancies diagnosed by the Oncology Department of Integrated Traditional Chinese and Western Medicine of China-Japan Friendship Hospital and treated with moderately/highly emetogenic chemotherapy were randomly divided into an experimental group （<italic>n</italic>=70） and a control group （<italic>n</italic>=70） according to the Good Clinical Practice （GCP）. Participants were given routine antiemetic treatment once 20 minutes before chemotherapy （tropisetron 5 mg + dexamethasone 5 mg or methylprednisolone 40 mg）. On this basis， the patients in the experimental group were treated with external application of modified Xiangsha Liujunzi Tang for 1 week on the second day after chemotherapy （the selected points were Zhongwan， Neiguan and Zusanli， 6 hours a day， and the application was changed every day）， and the patients in the control group were applied with comfort patch at the same time. The gastrointestinal reaction grade， subset concentration of lymphocytes， and Karnofsky score in two groups before and one week after chemotherapy were recorded， and all data were analyzed by SPSS 19.0 statistical software. Result:The grades of chemotherapy-related gastrointestinal reaction in two groups after treatment decreased as compared with that before treatment （<italic>P</italic><0.05）. The experimental group was superior to the control group after treatment in terms of grade decrease （<italic>P</italic><0.05）. The total response rate of the experimental group was 81.43% （57/70）， higher than 62.86% （44/70） in the control group （<italic>χ</italic>²=9.73， <italic>P</italic><0.05）. Stratified analysis was performed on the experimental group. Compared with the conditions before treatment， the grade of gastrointestinal reaction in patients with esophageal cancer and colorectal cancer of the experimental group decreased after treatment （<italic>P</italic><0.05）， and that in patients of different genders decreased after treatment （<italic>P</italic><0.05）. The results of immune function showed that there was no significant difference in the concentrations of CD3⁺， CD4⁺， CD8⁺， and NK cell subsets of the control group before and after treatment， while the concentrations of CD3⁺， CD4⁺， and NK cell subsets of the experimental group were higher than those before treatment and even superior to those in the control group （<italic>P</italic><0.05）. The Karnofsky score of quality of life in the experimental group was higher than that in the control group， with quality of life improved （<italic>P</italic><0.05）. Conclusion:External application of modified Xiangsha Liujunzi can improve the gastrointestinal reaction，immune function， and quality of life of patients with gastrointestinal tumors after moderately/highly emetogenic chemotherapy.

Epimedium flavonoids protect neurons and synapses in the brain via activating NRG1/ErbB4 and BDNF/Fyn signaling pathways in a chronic cerebral hypoperfusion rat model.

Cerebral hypoperfusion is a common feature of cerebral small vascular disease (CSVD), which has been considered as one of the causes of cognitive decline in recent years. Epimedium flavonoids (EF) are the main ingredients extracted from Epimedium. The purpose of this study was to investigate the effects of EF on cognitive impairment, and the underlying mechanisms in rats with permanent occlusion of the bilateral common carotid artery (2VO). EF (50, 100, and 200 mg/kg) was intragastrically administered for 12 weeks starting 2 weeks after 2VO surgery. The results showed that EF treatment improved learning and memory impairment in 2VO rats evaluated by novel object recognition and Y-maze tests. NeuN immunohistochemical staining indicated that EF alleviated neuronal loss in the hippocampus and cerebral cortex of 2VO rats. MAP-2 immunofluorescence staining and western blotting showed that EF protected neuronal dendrites and increased the expression of cytoskeleton proteins MAP-2 and NF200 in the hippocampus of 2VO rats. Moreover, EF protected the synapse ultrastructure detected by transmission electron microscopy, and increased the expression of synaptic plasticity-related proteins, including synaptophysin, synaptotagmin-I, synapsin I, PSD-95, p-NMDA2B, and p-CaMKII-α in the hippocampus of 2VO rats. In addition, EF increased the expression of neuregulin-1 (NRG-1), p-ErbB4, brain-derived neurotrophic factor (BDNF), p-Fyn, PI3K, p-Akt, and p-CREB in the hippocampus of 2VO rats. These results suggest that EF may protect neurons and synapses by activating the NRG1/ErbB4, BDNF/Fyn, and P13 K/Akt/CREB pathways in the hippocampus and cerebral cortex, thus improving cognitive impairment induced by chronic cerebral hypoperfusion. EF may be a potential candidate drug for chronic cerebral hypoperfusion and CSVD therapy.

Epimedium flavonoids improve cognitive impairment and white matter lesions induced by chronic cerebral hypoperfusion through inhibiting the Lingo-1/Fyn/ROCK pathway and activating the BDNF/NRG1/PI3K pathway in rats.

Chronic cerebral hypoperfusion is a common cause of cerebral small vascular disease (CSVD). White matter (WM) lesions are the typical pathological manifestation of CSVD and contribute to cognitive decline. Epimedium flavonoids (EF) are the main component in Epimedium brevicornu Maxim., which is commonly used in traditional Chinese medicine. The purpose of this study was to investigate the effects of EF on cognitive impairment and the underlying mechanisms in a CSVD rat model induced with chronic cerebral hypoperfusion. The model was established by permanent bilateral common carotid artery occlusion (2VO) in rats. EF (50, 100, and 200 mg/kg) was intragastrically administered once a day for 12 weeks starting 2 weeks after 2VO surgery. The learning and memory capacity of the rats were measured using the Morris water maze and step-through tests. WM lesions were observed by MRI-diffusion tensor imaging, transmission electron microscopy, and LFB staining. Oligodendrocytes were detected by immunohistochemistry. Western blotting assay was used to determine the level of protein expression. The results showed that EF significantly improved learning and memory impairment, alleviated WM nerve fiber injuries and demyelination, and increased the number of mature oligodendrocytes in the corpus callosum, subcortical WM, and periventricular WM in 2VO rats. Mechanistically, EF reduced the expression of Lingo-1 and ROCK2 and increased the levels of phosphorylated (p-) Fyn, brain-derived neurotrophic factor (BDNF), TrkB, neuregulin-1 (NRG-1), p-ErbB4, PI3K p85 and p110α, p-Akt, and p-CREB in the corpus callosum of 2VO rats. These results suggest that EF may improve cognitive impairment and WM lesions induced by chronic cerebral hypoperfusion through inhibiting the Lingo-1/Fyn/ROCK pathway and activating the BDNF/TrkB, NRG-1/ErbB4, and the downstream PI3K/Akt/CREB pathways in WM. Thus, EF can be used as a potential neuroprotective agent in CSVD therapy.

Shenqi Xingnao Granules ameliorates cognitive impairments and Alzheimer's disease-like pathologies in APP/PS1 mouse model.

Objective: Alzheimer's disease (AD) is along with cognitive decline due to amyloid-ß (Aß) plaques, tau hyperphosphorylation, and neuron loss. Shenqi Xingnao Granules (SQXN), a traditional Chinese medicine, significantly ameliorated the cognitive function and daily living abilities of patients with AD. However, till date, no study has investigated the mechanism of action of SQXN on AD. The present study aimed to verify the effects of SQXN treatment on cognitive impairments and AD-like pathologies in APP/PS1 mice. Methods: Four-month-old APP/PS1 transgenic (Tg) mice were randomly divided into a model group and SQXN-treated (3.5, 7, 14 g/kg per day) groups. Learning-memory abilities were determined by Morris water maze and object recognition test. All mice were sacrificed and the brain samples were collected after 75 d. The soluble Aß contents were detected by Elisa kit; The levels of expression of NeuN, APP, phosphorylated tau and related protein were measured by Western blotting; The inflammation factors were detected by the proinflammatory panel kit. Results: Four-month-old APP/PS1 mice were administered SQXN by oral gavage for 2.5 months. Using the Morris water maze tests and Novel object recognition, we found that SQXN restored behavioral deficits in the experimental group of Tg mice when compared with the controls. SQXN also inhibited neuronal loss (NeuN marker). SQXN treatment decreased soluble Aß42 through inhibiting the expression of sAPPß and BACE-1 without regulating full-length amyloid precursor protein (FL APP). Insulin degrading enzyme (IDE), the Aß degrading enzyme, were increased by SQXN. In addition, SQXN reduced hyperphosphorylated tau protein levels and prevented excessive activation of p-GSK-3ß in the brain of APP/PS1 mice. Compared with APP/PS1 transgenic negative mice, IFN-γ, IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-12p70, KC/GRO and TNF-α were not obviously changed in the brain of 6.5-month-old APP/PS1 transgenic (Tg) mice. However, SQXN could inhibited the expression of IL-2. Conclusion: These results demonstrate that SQXN ameliorates the cognitive impairments in APP/PS1 mice. The possible mechanisms involve its inhibition of neuronal loss, soluble Aß deposition, tau hyperphosphorylation and inflammation.

Cornel Iridoid Glycoside Suppresses Hyperactivity Phenotype in rTg4510 Mice through Reducing Tau Pathology and Improving Synaptic Dysfunction.

rTg4510 mice are transgenic mice expressing P301L mutant tau and have been developed as an animal model of tauopathies including Alzheimer's disease (AD). Besides cognitive impairments, rTg4510 mice also show abnormal hyperactivity behavior. Cornel iridoid glycoside (CIG) is an active ingredient extracted from Cornus officinalis, a traditional Chinese herb. The purpose of the present study was to investigate the effects of CIG on the emotional disorders such as hyperactivity, and related mechanisms. The emotional hyperactivity was detected by locomotor activity test and Y maze test. Immunofluorescent and immunohistochemical analyses were conducted to measure neuron loss and phosphorylated tau. Western blotting was used to detect the expression of related proteins. The results showed that intragastric administration of CIG for 3 months decreased the hyperactivity phenotype, prevented neuronal loss, reduced tau hyperphosphorylation and aggregation in the amygdala of rTg4510 mice. Meanwhile, CIG alleviated the synaptic dysfunction by increasing the expression of N-methyl-D-aspartate receptors (NMDARs) subunits GluN1 and GluN2A and αamino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) subunits GluA1 and GluA2, and increased the level of phosphorylated Ca2+/calmodulin dependent protein kinase II α (p-CaMK IIα) in the brain of rTg4510 mice. In conclusion, CIG may have potential to treat the emotional disorders in tauopathies such as AD through reducing tau pathology and improving synaptic dysfunction.

[Effect of Scutellariae Radix on expression of inflammatory cytokine protein and gene in lung of mice with viral pneumonia caused by influenza virus FM1 infection].

Mice models of viral pneumonia were induced by pulmonary adaptive strain FM1 of influenza A virus in Asian mice.RT-PCR and immunohistochemistry were used to dynamically observe the effect of Scutellariae Radix on the protein and gene expression of inflammatory cytokine in the lungs of the model mice infected by influenza virus FM1 at different phases. The partial mechanism of Scutellariae Radix in repairing the immune inflammatory damage of target organs of pneumonia caused by influenza virus was further explored. The results showed that Scutellariae Radix reduced protein and gene expression of proinflammatory cytokines tumor necrosis factor( TNF-α),interleukin IL-1,IL-6 in lung tissues from 3 rd to 5 th day after infection,and increased protein and gene expression of IL-10 and IFN-γ in lung tissues on the 5 th day after infection. Scutellariae Radix may inhibit excessive release of pro-inflammatory cytokines and promote the expression of anti-inflammatory cytokines,thereby inhibiting the systemic inflammatory response syndrome,reducing the immunoinflammatory pathological damage of lung caused by influenza virus FM1 infection,and promoting lung repair of tissue inflammatory lesions.

[The Extract from Punica Granatum (Pomegranate) Leaves Promotes Apoptosis and Impairs Metastasis in Prostate Cancer Cells].

OBJECTIVE: To investigate the effects of pomegranate leaves extract(PLE)on proliferation,apoptosis and metastasis of prostate cancer cells. METHODS: The proliferation of TRAMP-C1,DU145,PC3 prostate cancer cells treated with different concentrations of PLE (final mass concentrations were 12.5,25,50,100, 200 µg/mL,respectively) for different time (24,48,72 h) was detected by MTT assay. Colony formation assay was performed to verify the long-term effects of PLE on the proliferation of DU145 and PC3 cells.After being treated with PLE for 48 h,Hoechst-33258 staining was used to observe the changes in the nucleus,the cell apoptotic rate was detected by flow cytometry,and wound-healing migration assay was perform to test the change of migration. RESULTS: In comparison with the control group,PLE in the range of 12.5-200 µg/mL had a certain inhibitory effect on the proliferation of TRAMP-C1,DU145 and PC3 cells ( P<0.05).In the range of 6.25-100 µg/mL,the number of colony formation of DU145 and PC3 was significantly reduced( P<0.01).After PLE treated for 48 h, the apoptotic features of nuclear fragmentation and the formation apoptotic body was observed in PC3. With the increase of concentration,the apoptotic rate increased gradually ( P<0.05),and the ability of cells to migrate to the scratch area was significantly weaker than the control group ( P<0.01). CONCLUSION: PLE has effect on proliferation,apoptosis and metastasis of prostate cancer cells.

Screening, identification and activity evaluation of pancreatic lipase inhibition in Prunella vulgaris / 中国中药杂志

Pancreatic lipase (PL) inhibitors were firstly screened from Prunella vulgaris with PL immobilized on carboxylic acid-terminated magnetic nanoparticles, then these possible inhibitors were identified by LC-MS/MS and mixed standards. Finally, their inhibitory effects and types on PL were tested by p-nitrophenol method. The results showed that four PL inhibitors were screened out from P. vulgaris and confirmed by LC-MS/MS and mixed standards. The IC₅₈ and inhibition types were as follows: caffeic acid [(252.3±3.6) mg·L⁻¹, anti-competitive inhibition], rutin [(91.2±1.6)mg·L⁻¹, competitive inhibition], hesperidin [(31.5±4.4) mg·L⁻¹, competitive inhibition] and ursolic acid [(41.3±2.2) mg·L⁻¹, competitive inhibition]. Their inhibitive types and abilities on PL were related to their molecular size, hydrophobicity and the number of hydrogen bond with PL triplet.

Cornel Iridoid Glycoside Improves Locomotor Impairment and Decreases Spinal Cord Damage in Rats.

Purpose. This study was to investigate the effects of cornel iridoid glycoside (CIG) on spinal cord injury (SCI) in rats. Methods. The thoracic cord (at T9) of rats was injured by clip compression for 30 sec. Locomotor function was assessed using the Basso, Beattie, and Bresnahan (BBB) rating scale. Neuroanatomic stereological parameters as well as Nogo-A, p75 neurotrophin receptor (p75NTR), and ROCKII expression were measured by histological processing, immunohistochemistry, and stereological analyses. The axons passing through the lesion site were detected by BDA tracing. Results. Intragastric administration of CIG (60 and 180 mg/kg) improved the locomotor impairment at 10, 17, 24, and 31 days post-injury (dpi) compared with untreated SCI model rats. CIG treatment decreased the volume of the lesion epicenter (LEp) and increased the volume of spared tissue and the number of surviving neurons in the injured spinal cord at 31 dpi. CIG promoted the growth of BDA-positive axons and their passage through the lesion site and decreased the expression of Nogo-A, p75NTR, and ROCKII both in and around the LEp. Conclusion. CIG improved the locomotor impairment, decreased tissue damage, and downregulated the myelin-associated inhibition signaling pathway in SCI rats. The results suggest that CIG may be beneficial for SCI therapy.

Cornel iridoid glycoside reduces infarct size measured by magnetic resonance imaging and improves neurological function after focal cerebral ischemia in rats.

OBJECTIVE: To investigate the effect of cornel iridoid glycoside (CIG), an ingredient extracted from traditional Chinese herb Cornus offificinalis, on neurological function and infarct size in rats as measured by magnetic resonance imaging (MRI) after ischemic stroke. METHODS: Sprague-Dawley rats were divided into three group: control (n=11), model (n=20) and CIG (n=16) groups. Rats in the model and CIG groups underwent 90-min middle cerebral artery occlusion (MCAO) followed by reperfusion. Their neurological defect was measured by using a modified neurological severity score (mNSS). T2-weighted MRI (T2-MRI) of the brain was performed in vivo from 2 to 28 days after MCAO. The infarct volume in the brain was also measured using 2,3,5-triphenyltetrazolium chloride (TTC) staining 28 days after stroke. RESULTS: CIG, 60 mg/(kg day), administered by oral gavage starting from 6 h after the onset of MCAO improved neurological function at 7, 14, 21, and 28 days post occlusion (P<0.05 orP<0.01) and decreased mortality. The infarct volumes computed from the T2-MR images were reduced in the CIG-treated group compared with the model group at 7, 14 and 28 days after MCAO (P<0.05); and the rate at which the infarct volume decreased from 2 to 28 days was higher in the CIG-treated group than that in the model group (P<0.05). The infarct volumes measured by TTC staining were also decreased 28 days after stroke (P<0.05). CONCLUSION: CIG treatment, starting from 6 h after MCAO, reduced infarct size in the brain as measured by MRI and improved neurological function 2-28 days after focal cerebral ischemia in rats, suggesting that CIG could be a clinical application in improving stroke treatment.

Discovery, synthesis and biological evaluation of 2-(4-(N-phenethylsulfamoyl)phenoxy)acetamides (SAPAs) as novel sphingomyelin synthase 1 inhibitors.

Sphingomyelin synthase (SMS) has been proved to be a potential drug target for the treatment of atherosclerosis. However, few SMS inhibitors have been reported. In this paper, structure-based virtual screening was performed on hSMS1. SAPA 1a was discovered as a novel SMS1 inhibitor with an IC50 value of 5.2 µM in enzymatic assay. A series of 2-(4-(N-phenethylsulfamoyl)phenoxy)acetamides (SAPAs) were synthesized and their biological activities toward SMS1 were evaluated. Among them, SAPA 1j was found to be the most potent SMS1 inhibitor with an IC50 value of 2.1 µM in in vitro assay. The molecular docking studies suggested the interaction modes of SMS1 inhibitors and PC with the active site of SMS1. Site-directed mutagenesis validated the involvement of residues Arg342 and Tyr338 in enzymatic sphingomyelin production. The discovery of SAPA derivatives as a novel class of SMS1 inhibitors would advance the development of more effective SMS1 inhibitors.

Beneficial effects of the Chinese herbal medicine Sanjie Zhentong Capsule on experimental endometriosis in rats.

AIM: To analyze the composition of the Chinese herbal medicine Sanjie Zhentong Capsule (SJZTC) and test the therapeutic efficacy of each component in a rat model of endometriosis. METHODS: A rapid resolution liquid chromatography (RRLC) method coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS) has been developed for the analysis of SJZTC. Two main ingredients, Drac(h)onis sanguis and saponin, were tested in the endometriosis model. Sixty Lewis female rats were in the estrous cycle stage when endometriosis was experimentally initiated by peritoneal implantation of endometrial tissue. Four weeks later, a second laparotomy was performed and implant volumes were measured. After that, the implanted rats were randomized into five study groups: control group (treatment with saline), anastrozole group (treatment with anastrole, 18 µg per day), loureirin A group (treated with loureirin A, 97.2 mg), ginsenoside Re group (treated with ginsenoside Re, 64.8 mg), and SJZTC groups (treated with SJZTC, 86.4 mg) administered once a day for 4 weeks via gastric gavage. After four weeks of treatment, a third laparotomy was performed, implant volumes were re-measured, and the levels of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) were tested. RESULTS: A total of 38 compounds including, both the target and unknown compounds, were rapidly predicted in the capsule extract by the developed method. Compared with the control group, the anastrozole group, loureirin A group, ginsenoside Re group, and SJZTC treated group showed smaller implant volumes, as well as lower levels of VEGF and TNF-α in the peritoneal focus (P < 0.01 for all comparisons). Furthermore, parameters in the groups treated with SJZTC, loureirin A and ginsenoside Re were significantly better than the control group and the anastrozole group. These results indicate that SJZTC and its two main components are effective in reducing the development of endometriosis.

[Effects of Shenwu capsule on learning-memory ability and cholinergic function of brain in AD-like rat model induced by chronic infusion of sodium azide by minipump].

Because of the proposed importance of mitochondrial cytochrome C oxidase (COX) decrease in Alzheimer's disease (AD) , the protective effect of Shenwu capsule on mitochondrial deficiency model rats and its pharmacological mechanism were investigated in present study. Rats were administered with azide at 1 mg . kg-1 . h-1 subcutaneously via an Alzet minipump for 30 days. Tweny-four hours after the operation, the rats were administered intragastrically by Shenwu capsule with the dose of 0. 45, 0. 9 and 1. 8 g . kg-1 . d-1 for one month. Then learning-memory ability was determined by the watermaze test and passive avoidance tests. The activity of choline-acetyl-transfertase(ChAT) and acetylcholinesterase (AChE) in hippocampus and cortex of rats were measured by radiochemical method and hydroxylamine colorimetry separately. M-cholinergic receptor binding ability (M-binding) was assayed by radio binding. Chronic infusion of sodium azide via minipump induced learning-memory deficiency of rats. Both ChAT activity and M-binding decreased in hippocampus and cortex of model rats, however, the activity of AChE increased in hippocampus and was not affected at the cortex. As the result, the cholinergic function of the brain decreased in model rats. Shenwu capsule significantly improved learning and memory ability and the mechanism may be related with the improved cholinergic function in model brain: ChAT activity and M-binding significantly increased in Shenwu treated groups compared with model group; and the increased activity of AChE in hippocampus returned to normal. Mitochondria, especially mitochondrial cytochrome C oxidase, may play the key role in the early event of AD. Chronic, partial in vivo inhibition of mitochondrial cytochrome C oxidase in rats provides a suitable model mimicking several aspects of AD. Shenwu capsule indicate effectiveness in AD-like mitochondrial deficiency model rats, so it would be applied in the treatment of AD.

Cornel Iridoid Glycoside Attenuates Tau Hyperphosphorylation by Inhibition of PP2A Demethylation.

Aim. The aim of the present study was to investigate the effect of cornel iridoid glycoside (CIG) on tau hyperphosphorylation induced by wortmannin (WT) and GF-109203X (GFX) and the underlying mechanisms. Methods. Human neuroblastoma SK-N-SH cells were preincubated with CIG (50, 100, and 200 µg/ml, resp.) for 24 h and then exposed to 10 µM WT and 10 µM GFX for 3 h after washing out CIG. Immunohistochemistry was used to observe the microtubular cytoskeleton of the cultured cells. Western blotting was used to measure the phosphorylation level of tau protein, glycogen synthase kinase 3 ß (GSK-3 ß ), and protein phosphatase 2A (PP2A). The activity of PP2A was detected by a biochemical assay. Results. Preincubation of CIG significantly attenuated the WT/GFX-induced tau hyperphosphorylation at the sites of Thr205, Thr212, Ser214, Thr217, Ser396, and PHF-1 and improved the damage of morphology and microtubular cytoskeleton of the cells. CIG did not prevent the decrease in p-AKT-ser473 and p-GSK-3 ß -ser9 induced by WT/GFX. However, CIG significantly elevated the activity of PP2A by reducing the demethylation of PP2A catalytic subunit (PP2Ac) at Leu309 and the ratio of PME-1/LCMT in the WT/GFX-treated cells. The results suggest that CIG may be beneficial to the treatment of AD.

Determination and comparison of γ-aminobutyric acid (GABA) content in pu-erh and other types of Chinese tea.

Two previous studies have reported that pu-erh tea contains a high level of γ-aminobutyric acid (GABA), which is the major inhibitory neurotransmitter in the central nervous system and has several physiological functions. However, two other researchers have demonstrated that the GABA content of several pu-erh teas was low. Due to the high value and health benefits of GABA, analysis of mass-produced pu-erh tea is necessary to determine whether it is actually enriched with GABA. A high-performance liquid chromatography (HPLC) method was developed for the determination of GABA in tea, the results of which were verified by amino acid analysis using an Amino Acid Analyzer (AAA). A total of 114 samples of various types of Chinese tea, including 62 pu-erh teas, 13 green teas, 8 oolong teas, 8 black teas, 3 white teas, 4 GABA teas, and 16 process samples from two industrial fermentations of pu-erh tea (including the raw material and the first to seventh turnings), were analyzed using HPLC. Statistical analysis demonstrated that the GABA content in pu-erh tea was significantly lower than that in other types of tea (p < 0.05) and that the GABA content decreased during industrial fermentation of pu-erh tea (p < 0.05). This mass analysis and comparison suggested GABA was not a major bioactive constituent and resolved the disagreement GABA content in pu-erh tea. In addition, the GABA content in white tea was found to be significantly higher than that in the other types of tea (p < 0.05), leading to the possibility of producing GABA-enriched white tea.