Highly oxygenated germacrane-type sesquiterpenoids from the whole plant of Salvia cavaleriei H.Lév. and their biological activities.

The entire plant Salvia cavaleriei H.Lév. (Lamiaceae) is used as a traditional Chinese herbal medicine. Its leaves are edible, and the flowers can be soaked in water to make a health-care tea. In an effort to find natural bioactive chemical components, twelve undescribed germacrane-type sesquiterpenoids, salcavalins A-L, were isolated from the whole plant of S. cavaleriei and were identified as analogs. This is the first study to isolate highly oxygenated germacrane-type sesquiterpenoids from this plant. The structures of these undescribed compounds were elucidated by various spectroscopic methods, and their absolute configurations were confirmed by single-crystal X-ray diffraction analysis with Cu Kα radiation and electronic circular dichroism calculations. The biological activity of these undescribed compounds on the production of tumor necrosis factor-alpha in lipopolysaccharide induced NR8383 cells was evaluated, and salcavalins I and K showed anti-inflammatory activity to some extent. Salcavalins A-C, F and L were found to be neuroprotective with antiparkinsonic potential in a nematode (Caenorhabditis elegans) model. In addition, salcavalins F and I displayed marked phytotoxic activity against radish seeds at a low concentration of 50 ppm. Our findings provide scientific justification to show that bioactive sesquiterpenoids from the edible herb have anti-inflammatory in vitro, neuroprotective and phytotoxic activities.

Effect of ß-glucosidase on the aroma of liquid-fermented black tea juice as an ingredient for tea-based beverages.

The effects of ß-glucosidase on the volatile profiles and aroma stability of black tea juice were evaluated using gas-chromatography-mass spectrometry coupled with sensory analysis. During liquid fermentation of tea leaves, the addition of ß-glucosidase increased the concentration of aldehydes, strengthening the undesirable "green grassy" odour. However, the "green grassy" odour was counteracted by adding green tea extract during fermentation. At the same time, "flowery" flavour notes were enhanced, improving the overall aroma quality and strengthening the characteristic "sweet" aroma of black tea. Increased addition of ß-glucosidase released more free aroma alcohols from their glucosides. Two "fruity" and "floral" aroma components, benzyl alcohol and phenylethyl alcohol, were not significantly affected by heat treatment (95 °C water bath) and the overall aroma stability was not significantly affected by ß-glucosidase treatment. ß-Glucosidase treatment improved the aroma, colour and overall suitability of fermented black tea juice as an ingredient for tea-based beverages.

Antimicrobial diterpenoids from the leaves and twigs of Croton kongensis Gagnepain.

Croton kongensis Gagnepain. belongs to the genus Croton, the Euphorbiaceae family, mainly distributed in Hainan and southern Yunnan, China. The aim of present study was to acquire secondary metabolites of the ethanol extract obtained from the leaves and twigs of C. kongensis. Three new abietane-type diterpenoids, crokongenolides A-C (1-3), together with seven known diterpenoids (4-10), were isolated from the leaves and twigs of C. kongensis. The structures of the new compounds were determined by extensive spectroscopic methods (1D and 2D NMR, IR, and HRESIMS), and their absolute configurations were confirmed by single-crystal X-ray diffraction analysis or electronic circular dichroism (ECD) calculations. The absolute configuration of 4 was determined for the first time by single-crystal X-ray diffraction analysis with Cu-Kα irradiation. Some compounds were evaluated for their antimicrobial properties by assessing their inhibitory effects on Staphylococcus aureus, Candida albicans, and Escherichia coli. Compound 10 showed significant antimicrobial activity against S. aureus with MIC value of 1.56 µg/ml.

Chitosan Activated with Genipin: A Nontoxic Natural Carrier for Tannase Immobilization and Its Application in Enhancing Biological Activities of Tea Extract.

In this work, a non-toxic chitosan-based carrier was constructed via genipin activation and applied for the immobilization of tannase. The immobilization carriers and immobilized tannase were characterized using Fourier transform infrared spectroscopy and thermogravimetric analysis. Activation conditions (genipin concentration, activation temperature, activation pH and activation time) and immobilizations conditions (enzyme amount, immobilization time, immobilization temperature, immobilization pH, and shaking speed) were optimized. The activity and activity recovery rate of the immobilized tannase prepared using optimal activation and immobilization conditions reached 29.2 U/g and 53.6%, respectively. The immobilized tannase exhibited better environmental adaptability and stability. The immobilized tannase retained 20.1% of the initial activity after 12 cycles and retained 81.12% of residual activity after 30 days storage. The catechins composition analysis of tea extract indicated that the concentration of non-ester-type catechins, EGC and EC, were increased by 1758% and 807% after enzymatic treatment. Biological activity studies of tea extract revealed that tea extract treated with the immobilized tannase possessed higher antioxidant activity, higher inhibitory effect on α-amylase, and lower inhibitory effect on α-glucosidase. Our results demonstrate that chitosan activated with genipin could be an effective non-toxic carrier for tannase immobilization and enhancing biological activities of tea extract.

Sanweidoukou decoction, a Chinese herbal formula, ameliorates ß-amyloid protein-induced neuronal insult via modulating MAPK/NF-κB signaling pathways: Studies in vivo and in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional Chinese medicine Sanweidoukou decoction (DK-3) was a classical formula for the treatment of nervous system diseases, recorded in the Chinese medical classic Sibu Yidian. AIM OF THE STUDY: The present study is aim to investigate the neuroprotective effects of DK-3 on ß-amyloid (Aß) protein -induced AD-like pathologies and underlying molecular mechanisms both in vitro and in vivo studies. MATERIALS AND METHODS: Hydrolysates of DK-3 were analyzed by LC-ESI-MS/MS. In vitro, MTT was utilized to examine effects of DK-3 on Aß25-35-induced cytotoxicity in PC12 cells. In vivo, male Sprague-Dawley rats were administered with Aß25-35 to induce AD-like pathologies and behavioral evaluations were conducted via Morris water maze (MWM) test. Histopathological changes were observed by Hematoxylin-eosin (HE) straining. Immunohistochemistry (IHC) was used to detect the tau hyperphosphorylation at Thr181 site. The expression levels of tau hyperphosphorylation, inflammation-related cytokines such as COX-2, iNOS, TNF-α, IL-1ß, IL-6, the phosphorylated state of various mitogen-activated protein kinase (MAPK) signaling molecules (p38 MAPK, ERK, and JNK) and activation of nuclear factor κB (NF-κB) in vitro and in vivo were assessed via Western blot. RESULTS: In vitro, DK-3 dose-dependently increased cell viability of PC12 cells induced by Aß25-35. In vivo, DK-3 improved learning and memory abilities of Aß25-35-induced AD-like rats. Moreover, DK-3 reversed hyperphosphorylation of tau and reduced the production of inflammation-related cytokines through significantly inhibited MAPK and NF-κB signaling pathways both in vitro and in vivo studies. CONCLUSION: The present study suggested that the traditional Chinese medicine DK-3 may play a role in preventing and treating AD by reducing the hyperphosphorylation of tau protein and the expressions of inflammation-related cytokines via modulating the MAPK/NF-κB signaling pathways.

[Effect of electroacupuncture on morphine-conditioned place preference and activation of glutamatergic neurons in the ventromedial prefrontal cortex in rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli"(ST36) and "Sanyinjiao"(SP6) on conditioned place preference (CPP) and activation of glutamatergic neurons in the ventromedial prefrontal cortex (VMPFC) of morphine-addiction rats, so as to explore its mechanisms underlying detoxification. METHODS: Thirty male SD rats were randomly and equally divided into control, model and EA groups. The rats with acquisition of morphine-induced CPP received intraperitoneal injection of morphine (10 mg/kg) in the morphine-paired chamber, once daily for 3 consecutive days, and those of the control group received intraperitoneal injection of the same dose of normal saline in saline-paired chamber. Thirty minutes before CPP acquisition training, EA (2 Hz/100 Hz, 0.5 to 1.0 mA) was applied to ST36 and SP6 for 20 min every day. The double-labeled neurons of Fos/vesicular glutamate transporter 2 (VGLUT2) in the VMPFC were detected by using fluorescent immunohistochemistry. The discharges of the VMPFC neurons were recorded by using a multi-channel microarray electrophysiological system, followed by performing a z-score standardized processing. The ratio of firing rate frequency of rats in the morphine-paired chamber/saline-paired chamber was calculated, and further statistical analysis was conducted on the data based on the standardized z-scores. The neuronal firing characteristic of glutamatergic neuron is low frequency and wide wave. RESULTS: Compared with the control group, the score of morphine-induced CPP and numbers of Fos, VGLUT2-positive and Fos-VGLUT2 double-labeled positive cells were significantly increased in the model group (P<0.01，P<0.05). After EA and in comparison with the model group, the morphine CPP score and numbers of Fos, VGLUT2-positive and Fos-VGLUT2 double-labeled cells were significantly reduced in the EA group (P<0.01，P<0.05). The ratio of firing rate of the VMPFC neurons in the preference chamber and the percentage of inhibitory neurons as well as the z-score were considerably lower in the EA group than in the model group (P<0.001). CONCLUSION: EA can suppress morphine-induced seeking behavior in rats, which may be related to its inhibitory effect on glutamatergic neurons in the VMPFC.

Proteomic profiling uncovered the cytosolic superoxide dismutase BsSOD1 associated with plant defence in the herbal orchid Bletilla striata.

The herbal orchid Bletilla striata (Thunb.) Rchb.f. has a long cultivation history and has been widely used in medicines and cosmetics. The fungal infection leaf blight (LB) seriously threatens B. striata cultivation. Here, we systemically collected wild B. striata accessions and isolated the accessions with strong resistance against LB. We carried out proteomic profiling analysis of LB-resistant and LB-susceptible accessions, and identified a large number of differentially expressed proteins with significant gene ontology enrichment for 'oxidoreductase activity.' Of the proteins identified in the reactive oxygen species signalling pathway, the protein abundance of the Cu-Zn superoxide dismutase BsSOD1 and its gene expression level were higher in LB-resistant accessions than in LB-susceptible lines. Transient expression of the dismutase fused with yellow fluorescent protein determined that its subcellular localisation is in the cytoplasm. Our study provides new insights into the molecular markers associated with fungal infection in B. striata.

Thermostable Tannase from Aspergillus Niger and Its Application in the Enzymatic Extraction of Green Tea.

Tannase is widely used in tea beverage processing because of its ability to catalyze the hydrolysis of hydrolysable tannins or gallic acid esters and effectively improve the quality of tea extracts through enzymatic extraction. A new thermophilic tannase was cloned from Aspergillus niger FJ0118 and characterized. The tannase exhibited an optimal reaction temperature of 80 °C and retained 89.6% of the initial activity after incubation at 60 °C for 2 h. The enzymatic extraction of green tea at high temperature (70 °C) for a short time (40 min) was devised on the basis of the superior thermal stability of tannase. The enzymatic reaction significantly increased the total polyphenol content of green tea extract from 137 g·kg-1 to 291 g·kg-1. The enzymatic reaction effectively degraded the ester catechins into non-ester catechins compared with the water extraction method. Results suggested that the thermally stable tannase exhibited potential applications in the enzymatic extraction of green tea beverage.

[Brain regions responding to acupuncture stimulation of Zusanli (ST36) in healthy subjects analyzed on the basis of spontaneous brain activity].

OBJECTIVE: To determine the attributes of responses of the higher nerve center to acupuncture stimulation of Zusanli (ST36) on the basis of spontaneous brain activity, so as to explore the synchronization level of different brain rejoins after acupuncture. METHODS: All studies using fMRI to investigate the effect of acupuncture stimulation of ST36 and/or other acupoints on the human brain (at least 10 healthy subjects or patients in one group) published in journals from January of 1995 to January of 2018 were searched from databases of CNKI and PubMed by using keywords of acupuncture, electroacupuncture, and resting-state functional magnetic resonance images (rs-fMRI) or regional homogeneity (ReHo). Brain functional image data of acupuncture stimulation of ST36 and/or other acupoints were collected and analyzed with anisotropic effect size-signed differential mapping (AES-SDM) software (Meta-analysis), and those of acupuncture of simple ST36 analyzed as a subgroup. RESULTS: A total of 229 papers in Chinese and 109 in English were collected. According to our inclusive and exclusive standards, 11 papers containing 235 subjects were brought into analysis at last. Meta-analysis of brain image data of acupuncture at ST36 and/or other acupoints (comparison between pre- and post-acupuncture) revealed that the same brain regions (generality) which showed a significant increase in ReHo, are the right and left anterior cingulated gyrus, right caudate, left superior frontal gyrus, right middle frontal gyrus, and the right paracentral lobe, and those which showed a marked decrease of ReHo are the left mid-inferior occipital gyrus, left and right precentral and postcentral gyrus. The brain regions responding to acupuncture at ST36 only (specificity) are the right inferior parietal lobe, left middle inferior gyrus, right posterior lobe of cerebellum, and the left angular gyrus which displayed an increase of ReHo, and the right middle superior frontal gyrus which showed a decrease in ReHo. CONCLUSION: After acupuncturing at ST36, the relative generality and specificity of the central response in healthy subjects reflected as the location of the affected brain regions and the difference in the synchronization level of the corresponding spontaneous brain activities.

Brain regions responding to acupuncture stimulation of Zusanli (ST36) in healthy subjects analyzed on the basis of spontaneous brain activity / 针刺研究

OBJECTIVE: To determine the attributes of responses of the higher nerve center to acupuncture stimulation of Zusanli (ST36) on the basis of spontaneous brain activity, so as to explore the synchronization level of different brain rejoins after acupuncture. METHODS: All studies using fMRI to investigate the effect of acupuncture stimulation of ST36 and/or other acupoints on the human brain (at least 10 healthy subjects or patients in one group) published in journals from January of 1995 to January of 2018 were searched from databases of CNKI and PubMed by using keywords of acupuncture, electroacupuncture, and resting-state functional magnetic resonance images (rs-fMRI) or regional homogeneity (ReHo). Brain functional image data of acupuncture stimulation of ST36 and/or other acupoints were collected and analyzed with anisotropic effect size-signed differential mapping (AES-SDM) software (Meta-analysis), and those of acupuncture of simple ST36 analyzed as a subgroup. RESULTS: A total of 229 papers in Chinese and 109 in English were collected. According to our inclusive and exclusive standards, 11 papers containing 235 subjects were brought into analysis at last. Meta-analysis of brain image data of acupuncture at ST36 and/or other acupoints (comparison between pre- and post-acupuncture) revealed that the same brain regions (generality) which showed a significant increase in ReHo, are the right and left anterior cingulated gyrus, right caudate, left superior frontal gyrus, right middle frontal gyrus, and the right paracentral lobe, and those which showed a marked decrease of ReHo are the left mid-inferior occipital gyrus, left and right precentral and postcentral gyrus. The brain regions responding to acupuncture at ST36 only (specificity) are the right inferior parietal lobe, left middle inferior gyrus, right posterior lobe of cerebellum, and the left angular gyrus which displayed an increase of ReHo, and the right middle superior frontal gyrus which showed a decrease in ReHo. CONCLUSION: After acupuncturing at ST36, the relative generality and specificity of the central response in healthy subjects reflected as the location of the affected brain regions and the difference in the synchronization level of the corresponding spontaneous brain activities.

Eucheuma cottonii Sulfated Oligosaccharides Decrease Food Allergic Responses in Animal Models by Up-regulating Regulatory T (Treg) Cells.

In the present study, the anti-food allergy activity of Eucheuma cottonii sulfated oligosaccharide (ESO) was investigated. ESO was obtained by enzymatic degradation and purified by column chromatography. RBL-2H3 cells and BALB/c mouse model were used to test the anti-food allergy activity of ESO. The effects of ESO on the regulatory T (Treg) cells and bone marrow-derived mast cells (BMMCs) were investigated by flow cytometry. The results of in vivo assay showed that ESO decreased the levels of mast cell protease-1 and histamine and inhibited the levels of specific IgE by 77.7%. In addition, the production of interleukin (IL)-4 and IL-13 was diminished in the ESO groups compared to the non-ESO-treated group. Furthermore, ESO could up-regulate Treg cells by 22.2-97.1%. In conclusion, ESO decreased the allergy response in mice by reducing basophil degranulation, up-regulating Treg cells via Forkhead box protein 3 (Foxp3), and releasing IL-10. ESO may have preventive and therapeutic potential in allergic disease.

Investigation of sunlight-induced deterioration of aroma of pummelo (Citrus maxima) essential oil.

Deterioration of aromas of pummelo essential oil (EO) induced by sunlight was compared to those induced by heat and oxygen exposure using the techniques of sensory evaluation and GC-MS analysis. The sunlight-exposed EO was found to possess an oily off-flavor odor, which was significantly different from its counterparts induced by oxygen and heat. The strong oily note of the sunlight-exposed EO was attributed to the existence of linalool oxides and limonene oxides, as well as the lack of neral and geranial, for which UV sunlight was revealed to be the critical contributor causing the chemical reactions for the aroma changes. The results demonstrated that UV sunlight could significantly affect the aroma of the pummelo EO, providing valuable information that will benefit the production and storage of EO-based aromatic products.

[Transurethral resection of the prostate for urinary retention after seed implantation for prostate cancer: report of 2 cases].

OBJECTIVE: To investigate the effect of transurethral resection of the prostate (TURP) in the treatment of urinary retention following seed implantation for prostate cancer. METHODS: We treated 2 cases of urinary retention following seed implantation for prostate cancer by TURP, and analyzed their clinical data. RESULTS: The operations were successful in both of the cases, with mean operation time of 60 min. Postoperative nocturnal urination of the patients averaged 2 times per night as compared with 5 - 6 times per night preoperatively, and their scores on IPSS and quality of life were significantly improved after surgery. CONCLUSION: TURP affords satisfactory results in the treatment of urinary retention following seed implantation for prostate cancer patients.

8,8-Dimethyldihydroberberine with improved bioavailability and oral efficacy on obese and diabetic mouse models.

The clinical use of the natural alkaloid berberine (BBR) as an antidiabetic reagent is limited by its poor bioavailability. Our previous work demonstrated that dihydroberberine (dhBBR) has enhanced bioavailability and in vivo efficacy compared with berberine. Here we synthesized the 8,8-dimethyldihydroberberine (Di-Me) with improved stability, and bioavailability over dhBBR. Similar to BBR and dhBBR, Di-Me inhibited mitochondria respiration, increased AMP:ATP ratio, activated AMPK and stimulated glucose uptake in L6 myotubes. In diet-induced obese (DIO) mice, Di-Me counteracted the increased adiposity, tissue triglyceride accumulation and insulin resistance, and improved glucose tolerance at a dosage of 15mg/kg. Administered to db/db mice with a dosage of 50mg/kg, Di-Me effectively reduced random fed and fasting blood glucose, improved glucose tolerance, alleviated insulin resistance and reduced plasma triglycerides, with better efficacy than dhBBR at the same dosage. Our work highlights the importance of dihydroberberine analogs as potential therapeutic reagents for type 2 diabetes treatment.

[Analysis of growth dynamics of Curcuma wenyujin].

OBJECTIVE: To study the accumulation of matter and dynamic of chemical compositions in rhizoma of Curcuma wenyujin for its cultivation. METHODS: The weight of various parts and chemical composition content of rhizoma were determined at regular time. RESULTS: The growth of Curcuma wenyujin could be divided into the seedling period, the rhizoma growing period and dry matter accumulating period. In seedling period, the aerial parts were the center of organism, the matter accumulated mainly in leaves; In rhizoma growing period, the organic center were gradually transformed from the aerial parts to the underground parts. The tendency of total chemical composition quantity in rhizome presented rises at first but gradually dropped at last. CONCLUSIONS: The growth of Curcuma wenyujin has two organic centers, the seedling period and the rhizoma vegetal and weighted period. The total chemical composition quantity in Curcuma wenyujin rhizome reaches the maximum value from the last third part of October to mid-November.

Targeting the delivery of glycan-based paclitaxel prodrugs to cancer cells via glucose transporters.

This report describes the synthesis of four novel paclitaxel based prodrugs with glycan conjugation (1-4). Glycans were conjugated using an ester or ether bond as the linker between 2'-paclitaxel and the 2'-glucose or glucuronic acid moiety. These prodrugs showed good water solubility and selective cytotoxicity against cancer cell lines, but showed reduced toxicity toward normal cell lines and cancer cell lines with low expression levels of GLUTs. The ester conjugated prodrug 1 showed the most cytotoxicity among the prodrugs examined and could be transported into cells via GLUTs. Fluorescent and confocal microscopy demonstrated that targeted cells exhibited morphological changes in tubulin and chromosomal alterations that were similar to those observed with paclitaxel treatment. Therefore, these glycan-based prodrugs may be good drug candidates for cancer therapy, and the glycan conjugation approach is an alternative method to enhance the targeted delivery of other drugs to cancer cells that overexpress GLUTs.

[A prospective study of combined chemoradiotherapy followed by surgery in the treatment of esophageal carcinoma].

OBJECTIVE: To evaluate the effect of combined chemoradiotherapy followed by surgery for patients with esophageal carcinoma. METHODS: Ninety-seven patients with stage II or III esophageal carcinoma without contraindication against operation and chemoradiotherapy, were randomly divided into two groups: combined group (Group A) 48 and control group (Group B) 49. Patients in group A were given neoadjuvant treatment consisted of chemotherapy with 5-fluorouracil and cisplatin for 2 cycles and radiotherapy of DT36 Gy/12 f/17 d. Three weeks later, operation was performed. Patients in group B were given operation alone. Survival rate was calculated with Kaplan-Meier method. Chi and Log-rank test was used to assess the difference between the two groups. RESULTS: The radical resectability of group A and group B were 85.4% and 65.3% (P = 0.018 1). The lymph node metastasis rate of the two groups were 21.7% and 45.7% (P = 0.019 4). The T stage of group A was significantly lowered (P = 0.003 6). The local and regional recurrence rate of two groups were 34.8% and 58.7% (P = 0.023 6), while there was no significant difference in operative complications between the two groups. Significant improvement in the long-term survival rate was observed in group A, especially in patients who achieved partial and complete response with high 5-year survival rate of 56.5%. CONCLUSION: Preoperative neoadjuvant chemoradiotherapy is able to reduce the tumor and tumor stage, lower the lymph node metastasis rate and local or regional recurrence rate, also it can improve radical resectability and long-term survival without increasing the operative complications.