HS-SPME coupled with GC-MS for elucidating differences between the volatile components in wild and cultivated Atractylodes chinensis.

INTRODUCTION: Atractylodes chinensis is a Chinese herb that is used in traditional medicine; it contains volatile components that have enormous potential for pharmaceutical, food, and cosmetic applications. The destruction of wild resources demands significant improvement in the quality of artificial cultivation of Atractylodes chinensis. However, little is known about the compositional differences in the volatile substances derived from the wild and cultivated varieties of Atractylodes chinensis. OBJECTIVES: We aimed to evaluate the specific components of Atractylodes chinensis and analyse the similarities and differences between the volatile components and metabolic pathways in the wild and cultivated varieties. MATERIAL AND METHODS: Metabolomic analysis using gas chromatography-mass spectrometry (GC-MS) was employed following the extraction of volatile components from Atractylodes chinensis using headspace solid-phase microextraction (HS-SPME). RESULTS: A total of 167 volatile metabolites were extracted, and 137 substances were matched with NIST and Wiley databases. Among them, 76 compounds exhibited significant differences between the two sources; these mainly included terpenes, aromatics, and esters. KEGG enrichment analysis indicated that the differential metabolites were primarily involved in the biosynthesis of secondary metabolites, terpene biosynthesis, and limonene and pinene degradation; all these pathways have geranyl diphosphate (GDP) as the common link. CONCLUSION: The total content of volatile substances extracted from wild Atractylodes chinensis was 2.5 times higher than that from the cultured variety; however, each source had different dominant metabolites. This study underscores the necessity for protecting wild Atractylodes chinensis resources, while enhancing the quality of cultivated Atractylodes chinensis.

Discovery of enzymes to biotransform ginsenoside Rd into ginsenosides F2 and CK using metagenomics and genomic mining.

Ginsenosides are the main active components of ginseng, including many types and different contents. Among them, minor ginsenosides have better biological functions and pharmacological activities than those of the major ginsenosides. However, minor ginsenosides cannot be obtained in large quantities, but by means of enzymatic transformation technology, some major ginsenosides can be de-glycosylated at a specific position to generate minor ginsenosides. In this study, we report two glycosidase genes associated with the conversion of ginsenoside Rd to ginsenosides F2 or CK. SWMU-CK-1 was identified among the total genes extracted from the feces of plum deer by local Blast screening for putative ginsenoside conversion function, which could cause the conversion of ginsenoside Rd → F2 → CK. The other gene was found in the Bifidobacterium breve 689b SGAir 0764 chromosome genome, which might have the same function as the ß-glucosidase gene testified by the gene matching, named SWMU-F2-2, and can achieve the Rd → F2 transformation. This study reports two genes that enable achieving the biotransformation of rare ginsenosides, while it provides a new insight and a promising approach to explore new genes and develop new functions of existing genes.

Antioxidant and anti-inflammatory activity of constituents isolated from Dendrobium nobile (Lindl.).

Dendrobium nobile (Lindl.) have long been used as herbal tea and a traditional herbal medicine to treat Alzheimer's disease (AD). In the current study, nineteen compounds (1-19), including two new vitamin E homologues (1-2), one new sesquiterpene (6), and two new dendrobines (7, 8), were isolated and identified from stems of Dendrobium nobile. Their structures were elucidated on the basis of NMR, 13C NMR calculation, and DP4+ probability analyses. The absolute configurations of new compounds were determined by electronic circular dichroism (ECD) data analysis. Antioxidant, anti-inflammatory, and cytotoxic activities of isolated compounds were evaluated. Among them, compound 2 demonstrated significant antioxidant activity compared with ascorbic acid (VC), while compounds 2 and 4 also exhibited an equal effect to positive control cisplatin. This study on the biological activity of the new vitamin E homologues from Dendrobium nobile may indicate its potential application in the pharmaceutical and food industries.

Three main metabolites from Wolfiporia cocos (F. A. Wolf) Ryvarden & Gilb regulate the gut microbiota in mice: A comparative study using microbiome-metabolomics.

Wolfiporia cocos (F. A. Wolf) Ryvarden & Gilb, also known as Poria cocos is an ancient edible and medicinal mushroom that has been valued for thousands of years for its tranquilizing, diuretic, and spleen-enhancing properties. Because of the mushroom's complex composition, its pharmacological effects have not been fully clarified. Therefore, to expand our knowledge of these effects from a pharmacological perspective and exploit potential medicinal value of fungal mushroom, we extracted three main metabolites from P. cocos, including water-soluble polysaccharides (PCX), water-insoluble polysaccharides (PCY), and triterpenoid saponins (PCZ) for intragastric injection into mice. These injections were made to explore the component's effects on the mice's gut microbiota and their metabolomics. The microbiota analysis showed that PCY had the strongest effect on regulating gut microbiota through altering its composition and increasing the number of Lactobacillus (p < 0.01). A total of 1,828 metabolites were detected using metabolomics methods, and the results showed that the three main active metabolites of P. cocos significantly changed the content of short-chain peptides in intestinal metabolites. In conclusion, our study further investigated the pharmacological functions of P. cocos, and revealed the differing effects of its three main metabolites on gut microbiota. The results suggested that PCY is a prominent prebiotic, and provided us with new insights into the potential development of fungal polysaccharides in Chinese traditional medicine.

Characterization and prebiotic properties of pectin polysaccharide from Clausena lansium (Lour.) Skeels fruit.

Pectins have proven to be advantageous for human health as they regulate beneficial microbial communities and enhance immunity. The fruit of Clausena lansium (Lour.) Skeels (Wampee), also referred to as "treasure in fruit", is rich in pectin polysaccharides. In this study, a homogalacturonan-type pectin (CCP2) with a molecular weight of 8.9 × 104 Da and degree of esterification of 42.86% was isolated from Wampee fruit. The gut microbiota regulation and phagocytosis-enhancing properties of CCP2 were examined in vivo and in vitro, respectively. Oral administration of CCP2 dramatically decreased the abundance of Bacteroidetes and increased the abundance of Firmicutes in intestinal bacteria in mice. The content of short-chain fatty acids in the feces also significantly improved. Moreover, CCP2 exhibited excellent phagocytosis-enhancing activities on RAW 264.7 macrophages. These results suggested that CCP2 could be a potential gut microbiota regulator and phagocytosis-enhancer, which could be used in food products to promote health through beneficial manipulation of gut microbiota.

Metagenomic Analysis of Antibiotic Resistance Genes in Untreated Wastewater From Three Different Hospitals.

Controlling antibiotic resistance genes (ARGs) is a worldwide intervention to ensure global health. Hospital wastewater is the main pollution source of antibiotic-resistant bacteria and ARGs in the environment. Expanding our knowledge on the bacterial composition of hospital wastewater could help us to control infections in hospitals and decrease pathogen release into the environment. In this study, a high-throughput sequencing-based metagenomic approach was applied to investigate the community composition of bacteria and ARGs in untreated wastewater from three different types of hospitals [the general hospital, traditional Chinese medicine (TCM) hospital, and stomatology hospital]. In total, 130 phyla and 2,554 genera were identified from the microbiota of the wastewaters, with significantly different bacterial community compositions among the three hospitals. Total ARG analysis using the Antibiotic Resistance Genes Database (ARDB) and Comprehensive Antibiotic Resistance Database (CARD) revealed that the microbiota in the wastewaters from the three hospitals harbored different types and percentage of ARGs, and their composition was specific to the hospital type based on the correlation analysis between species and ARG abundance, some ARGs contributed to different bacterial genera with various relationships in different hospitals. In summary, our findings demonstrated a widespread occurrence of ARGs and ARG-harboring microbiota in untreated wastewaters of different hospitals, suggesting that protection measures should be applied to prevent human infections. Concurrently, hospital wastewater should be treated more specifically for the removal of pathogens before its discharge into the urban sewage system.

[Intervention mechanism of moxa-cone moxibustion at Shu- and Mu-acupoints on the expression of RORγt/Foxp3 in asthma mice: an analysis based on PI3K signaling pathway].

OBJECTIVE: To explore the effect of moxa-cone moxibustion at "Feishu" (BL13) and "Zhongfu" (LU1) on the contents of related inflammatory factors in serum and bronchoalveolar lavage fluid (BALF), and the expression of phosphatidy-linositol-3-kinase (PI3K), retinoic acid related orphan receptor γt (RORγt) and fork/wing helix transcription factor 3 (Foxp3) in lung tissue in asthma mice. METHODS: Sixty male Balb/c mice were divided into normal, model, LY294002 (an inhibitor of PI3K, LY), electroacupuncture (EA) and moxa-cone moxibustion (moxibustion) groups (n=12 in each group). The asthma model was established by intraperitoneal injection of ovalbumin sensitization. The mice in the LY group were injected with LY 294002 (7.5 mg/kg) via the tail vein. EA or moxa-cone moxibustion was applied at BL13 and LU1 in the EA or moxibustion group once daily for 2 weeks. The levels of immunoglobulin E (IgE), interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and BALF were detected by enzyme-linked immunosorbent assay. The expressions of PI3K, RORγt and Foxp3 in lung tissue were detected by real-time fluorescent quantitative polymerase chain reaction (QRT PCR) and immunohistochemistry. RESULTS: Compared with the normal group, the levels of IgE and IL-4 in serum and BALF, the expressions of PI3K and RORγt mRNA and protein in lung tissue were significantly increased (P<0.01), while the levels of IFN-γ, and the expressions of Foxp3 mRNA and protein were significantly reduced (P<0.01) in the model group. Compared with the model group, the levels of IgE and IL-4 in serum and BALF, the expressions of PI3K and RORγt mRNA and protein in lung tissue were significantly decreased (P<0.01, P<0.05), and the levels of IFN-γ in serum and BALF, the expressions of Foxp3 mRNA and protein in lung tissue were significantly increased (P<0.01) in the LY, EA and moxibustion groups. CONCLUSION: Moxa cone moxibustion at Shu- and Mu-acupoints of the lung meridian can reduce airway inflammatory reaction and control asthma attacks in asthma mice, which is closely related to its effects in regulating the expressions of RORγt and Foxp3 through PI3K signaling pathway.

The chain mediating role of social support and stigma in the relationship between mindfulness and psychological distress among Chinese lung cancer patients.

PURPOSE: Psychological distress greatly impaired the psychological and physical well-being of lung cancer patients. Identification of protective and risk factors is a prerequisite of developing effective psychological treatment protocol. The study aims to determine the relationship of mindfulness and psychological distress and further clarify the mechanism of mindfulness against psychological distress through perceived stigma and social support among Chinese lung cancer patients. METHOD: A cross-sectional survey study involving 441 valid Chinese lung cancer patients was conducted from September 2018 to August 2019. After all validated questionnaires that measured psychological distress, level of mindfulness, social support, and perceived stigma were returned by patients, we firstly performed correlation analysis to assess the associations between mindfulness, social support, perceived stigma, and psychological distress. Then structural equation modelling analysis was conducted to further clarify the mediating effects of perceived stigma and social support on the relationship between mindfulness and psychological distress. RESULTS: According to our hypothesis and further modification, our revised model adequately fits to data. Mindfulness (ß = - 0.107, p = 0.008) and social support (ß = - 0.513, p < 0.001) had a direct effect on psychological distress. Meanwhile, mindfulness had a direct effect on perceived stigma (ß = - 0.185, p < 0.001), and perceived stigma had a direct effect on social support (ß = - 0.373, p < 0.001). Furthermore, mindfulness had also the indirect effect on psychological distress through the chain mediating role of stigma and social support among lung cancer patients. CONCLUSIONS: Mindfulness has direct negative effect on psychological distress and has also indirectly negative psychological distress through impacting social support and perceived stigma.

Neuroprotective Effects of Safflower Flavonoid Extract in 6-Hydroxydopamine-Induced Model of Parkinson's Disease May Be Related to its Anti-Inflammatory Action.

Safflower (Carthamus tinctorius. L.), a Chinese materia medica, is widely used for the treatment of cardiovascular and cerebrovascular diseases, with flavonoids being the major active components. Multiple flavonoids in safflower bind to Parkinson's disease (PD)-related protein DJ-1. Safflower flavonoid extract (SAFE) improved behavioral indicators in a 6-hydroxydopamine (6-OHDA)-induced rat model of PD; however, the underlying mechanisms remain unclear. We used a 6-OHDA-induced mouse model of PD and a primary neuron-astrocyte coculture system to determine the neuroprotective effects and mechanisms of SAFE. After three weeks of SAFE administration, behavioral indicators of PD mice were improved. SAFE regulated the levels of tyrosine hydroxylase (TH) and dopamine metabolism. It significantly inhibited the activation of astrocytes surrounding the substantia nigra and reduced Iba-1 protein level in the striatum of PD mice. SAFE reduced the plasma content of inflammatory factors and suppressed the activation of nod-like receptor protein 3 (NLRP3) inflammasome. In the coculture system, kaempferol 3-O-rutinoside and anhydrosafflor yellow B significantly improved neuronal survival, suppressed neuronal apoptosis, and reduced IL-1ß and IL-10 levels in the medium. Thus, SAFE showed a significant anti-PD effect, which is mainly associated with flavonoid anti-inflammatory activities.

Extraction, purification, and determination of the gastroprotective activity of glucomannan from Bletilla striata.

In this study, a water-soluble polysaccharide (BSP) was extracted and purified from pseudobulb of Bletilla striata. The preliminary structure and gastroprotective activity of BSP were analyzed. Results indicate that BSP is a glucomannan with a molar ratio of 7.45:2.55 (Man:Glc), and its molecular weight is approximately 1.7 × 105 Da. BSP displayed outstanding protective action against ethanol-induced GES-1 cell injury in vitro, as well as, excellent gastroprotective activity in vivo. Especially, a high-dose of BSP (100 mg/kg) could reduce the ulcer index of the gastric mucosa and increase the percentage of ulcer inhibition, which possibly caused by enhancing the antioxidant capacity and inhibiting the apoptotic pathway in gastric tissue. Interestingly, BSP exhibited a comparative gastroprotective activity to that of positive control (omeprazole). In summary, our results indicated that BSP could be considered as a potential supplement for the prevention of gastric injury.

[Screening, identification and antagonistic effect evaluation of biocontrol strains to Asarum leaf blight].

Leaf blight is the main disease of Asarum. At present, chemical treatment is main measure for disease control, and there is no report on biological control. In order to achieve the biological control of Asarum leaf blight, the biocontrol strains with antagonistic effect on Asarum leaf blight were screened. The rhizosphere bacteria of healthy Asarum plants were isolated by soil dilution method, and the isolated strains were screened by the methods of antagonistic antifungal and fermentation liquid antifungal, then the strains were identified and the control effect in vivo was determined. Abiocontrol bacterial strains S2-31 which with high antagonism to leaf blight was obtained from more than 100 isolated strains. The inhibitory rates of antagonistic antifungal and fermentation liquid antifungal reached 92.47% and 60.56%, respectively. It was identified by morphology and 16 S rDNA sequence analysis, and the strain was identified as Brevibacillus laterosporus. The results of indoor potted experiment showed that the control effect was 79.87%, 71.44% and 66.82% on the 3 rd, 5 th and 7 th day after inoculation, respectively, which indicated that S2-31 could reduce the disease index and control the development of Asarum leaf blight.

[Effect of moxa-cone moxibustion at lung's back-shu points and front-mu points on Th17/Treg balance in mice with asthma].

OBJECTIVE: To observe the effect of moxa-cone moxibustion at lung's back-shu points and front-mu points on the expression of helper T lymphocyte 17 (Th 17)/regulatory T lymphocyte (Treg) in mice with asthma, and to explore the possible mechanism of moxa-cone moxibustion on asthma. METHODS: Sixty SPF-grade healthy male Balb/c mice were randomly divided into a normal group, a model group, an LY294002 group (LY group), an electroacupuncture (EA) group and a moxibustion group, 12 mice in each group. Asthma model was replicated by using ovalbumin (OVA) sensitization. Except the mice in the normal group, all the mice were intraperitoneally injected with sensitization solution (containing 15 µg of OVA and 30 mg of aluminum hydroxide) on the 1st day, 7th day and 14th day, 0.5 mL per mice; from the 15th day, 1% OVA solution was atomized for 20 min, once a day for 2 weeks; the mice in the normal group was treated with identical operations but with 0.9% sodium chloride solution. The mice in the LY group were treated with injection of LY294002 at tail vein on the 13th day, 14th day and 15th day. At the beginning of the 15th day, The mice in the EA group were treated with EA at "Feishu" (BL 13) and "Zhongfu" (LU 1) with disperse-dense wave, frequency of 2 Hz/20 Hz, intensity of 1 mA, 15 min each time, once a day for 2 weeks. The mice in the moxibustion group was treated with moxa-cone moxibustion at "Feishu" (BL 13) and "Zhongfu" (LU 1) from the 15th day, three moxa-cones per acupoint, once a day for 2 weeks. On the 16th day, 18th day and 22nd day, the incubation period of asthma was recorded. On the 29th day, all the samples were collected. The expressions of IL-17 and IL-10 in serum and bronchoalveolar lavage fluid (BALF) were detected by ELISA method. The pathological changes of lung tissue were observed by HE staining. The percentage of Th17, Treg and Th17/Treg ratio in spleen tissue were detected by flow cytometry method. RESULTS: Compared with the normal group, the incubation period of asthma in the model group was significantly shortened (P<0.01), while that in the LY group, the EA group and the moxibustion group was longer than that in the model group (P<0.01). After the intervention, bleeding and inflammatory exudation in the lung tissue were observed in the model group, and a large number of inflammatory cell infiltration around the bronchus and alveoli was found, and extensive consolidation appeared; the pathological changes of lung tissue in the LY group, the EA group and the moxibustion group were significantly reduced, and the structure of bronchus and alveoli was basically normal, and the inflammatory exudation and inflammatory cell infiltration were significantly reduced compared with the model group; the inflammatory reaction in the moxibustion group was slightly less than that in the LY group and the EA group. Compared with the normal group, the content of IL-17 in serum and BALF, the percentage of Th17 and Th17/Treg ratio in spleen tissue in the model group after intervention were increased (P<0.01), while the content of IL-10 in serum and BALF and the percentage of Treg in spleen tissue were decreased (P<0.01, P<0.05). Compared with the model group, the contents of IL-17 in serum and BALF, Th17 percentage and Th17/Treg ratio in spleen tissue in the LY group, the EA group and the moxibustion group were significantly reduced (P<0.01, P<0.05), while the content of IL-10 in serum and BALF and the percentage of Treg in spleen tissue were increased (P<0.01, P<0.05). The above indice had no significant difference among all the intervention groups (P>0.05). CONCLUSION: The Th17/Treg is imbalanced in asthmatic body. The moxibustion at lung's back-shu points and front-mu points can control asthma by regulating Th17/Treg imbalance and restoring immune homeostasis.

Liujunzi Tang, a famous traditional Chinese medicine, ameliorates cigarette smoke-induced mouse model of COPD.

ETHNOPHARMACOLOGICAL RELEVANCE: Liujunzi Tang is a traditional herbal medicine widely used in East Asia and clinically applied to treat Phlegm-Heat Syndrome. The purpose of the present study was to investigate the protective effects of Liujunzi Tang on cigarette smoke-induced (CS) mouse model of chronic obstructive pulmonary disease (COPD) and explore its potential molecular mechanism. MATERIALS AND METHODS: The mice received 1h of cigarette smoke for 8 weeks. The serum levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß and IL-6 were determined by enzyme-linked immunosorbent assay (ELISA) kits. Superoxide dismutase (SOD) and malondialdehyde (MDA) were tested by biochemical methods. Histopathological alteration was observed by hematoxylin-eosin (H&E) staining. Additionally, the expressions of nuclear transcription factor-κB (NF-κBp65) and (inhibitor of NF-κB)IκB-α were determined by western blot and immunohistochemistry analysis. RESULTS: Liujunzi Tang enhanced the activities of antioxidant enzymes and attenuated the levels of lipid oxidative production, meanwhile significantly inhibited the generations of inflammatory cytokines by inhibiting the phosphorylation of IκB-α and NF-κB. CONCLUSION: Our findings indicated that Liujunzi Tang exhibited the protective effect on cigarette smoke-induced COPD mice by anti-inflammatory and anti-oxidative properties through the inhibition of NF-κB activation.

Neuroprotective Effects of a Standardized Flavonoid Extract from Safflower against a Rotenone-Induced Rat Model of Parkinson's Disease.

Parkinson's disease (PD) is a major age-related neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra par compacta (SNpc). Rotenone is a neurotoxin that is routinely used to model PD to aid in understanding the mechanisms of neuronal death. Safflower (Carthamus tinctorius. L.) has long been used to treat cerebrovascular diseases in China. This plant contains flavonoids, which have been reported to be effective in models of neurodegenerative disease. We previously reported that kaempferol derivatives from safflower could bind DJ-1, a protein associated with PD, and that a flavonoid extract from safflower exhibited neuroprotective effects in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced mouse model of PD. In this study, a standardized safflower flavonoid extract (SAFE) was isolated from safflower and found to primarily contain flavonoids. The aim of the current study was to confirm the neuroprotective effects of SAFE in rotenone-induced Parkinson rats. The results showed that SAFE treatment increased body weight and improved rearing behavior and grip strength. SAFE (35 or 70 mg/kg/day) treatment reversed the decreased protein expression of tyrosine hydroxylase, dopamine transporter and DJ-1 and increased the levels of dopamine and its metabolite. In contrast, acetylcholine levels were decreased. SAFE treatment also led to partial inhibition of PD-associated changes in extracellular space diffusion parameters. These changes were detected using a magnetic resonance imaging (MRI) tracer-based method, which provides novel information regarding neuronal loss and astrocyte activation. Thus, our results indicate that SAFE represents a potential therapeutic herbal treatment for PD.

Neuroprotective Effects of A Standardized Flavonoid Extract of Safflower Against Neurotoxin-Induced Cellular and Animal Models of Parkinson's Disease.

Safflower has long been used to treat cerebrovascular diseases in China. We previously reported that kaempferol derivatives of safflower can bind DJ-1, a protein associated with Parkinson's disease (PD), and flavonoid extract of safflower exhibited neuroprotective effects in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced mouse model of PD. In this study, a standardized safflower flavonoid extract (SAFE) was isolated from safflower and mainly contained flavonoids. Two marker compounds of SAFE, kaempferol 3-O-rutinoside and anhydrosafflor yellow B, were proven to suppress microtubule destabilization and decreased cell area, respectively. We confirmed that SAFE in dripping pill form could improve behavioural performances in a 6-hydroxydopamine (6-OHDA)-induced rat model of PD, partially via the suppression of α-synuclein overexpression or aggregation, as well as the suppression of reactive astrogliosis. Using an MRI tracer-based method, we found that 6-OHDA could change extracellular space (ECS) diffusion parameters, including a decrease in tortuosity and the rate constant of clearance and an increase in the elimination half-life of the tracer in the 6-OHDA-lesioned substantia nigra. SAFE treatment could partially inhibit the changes in ECS diffusion parameters, which might provide some information about neuronal loss and astrocyte activation. Consequently, our results indicate that SAFE is a potential therapeutic herbal product for treatment of PD.

[Most advance on chemical and biological investigations of gorgonian-octocorals].

This review presents the most recent chemical and biological investigations on one of the marine invertebrates, gorgonian octocoral. It summarizes all 432 new compounds published in 2002-2009, which consists of 46 sesquiterpenoids (including 2 new natural products, NNP), 282 diterpenoids (including 4 from Pennatulacea octocoral and one artifact), 76 steroids, and 29 alkaloids or other types (2 NNP included). In this paper, according to the structure division, the new compounds are described in combination with the taxonomy of the investigated animals, and its simultaneous bioactivity results. Novel skeletons and complex structures are paid most emphasis on its features, means of structural elucidation and the proposed biogenesis pathway. The source of all new compounds and the different diterpenoid skeleton types are all listed and analyzed, as well as the commonly used Chinese names or some proposed ones for diterpenoid skeletons. Furthermore, this papers deals with all biological test toward the gorgonian new metabolites, i.e. anti-cancer, anti-inflammatory, anti-bacterial(against Staphylococcus aureus bacteria, Mycobacterium tuberculosis etc), anti-malaria, and anti-fouling as well, in which anti-cancer activity and cytotoxicity were additionally, a discussion and prospect are proposed regarding chemical overview on gorgonian. This review, hopefully, can be useful in proving data and references for further chemical and biological research of China sea gorgonian, for the studies on chemical ecology, and for the discovery of new and bioactive compounds or the marine-derived leading targets.

Shuanghuanglian injection downregulates nuclear factor-kappa B expression in mice with viral encephalitis.

A mouse model of viral encephalitis was induced by intracranial injection of a Coxsackie virus B3 suspension. Quantitative real-time reverse transcription-PCR and western blot assay were applied to detect mRNA and protein expression of intelectin-2 and nuclear factor-kappa B in the viral encephalitis and control groups. Nuclear factor-kappa B and intelectin-2 mRNA and protein expression were significantly increased in mice with viral encephalitis. After intraperitoneal injection of Shuanghuanglian at a dose of 1.5 mg/kg for 5 successive days, intelectin-2 and nuclear factor-kappa B protein and mRNA expression were significantly decreased. To elucidate the relationship between intelectin-2 and nuclear factor-kappa B, mice with viral encephalitis were administered an intracerebral injection of 107 pfu recombinant lentivirus expressing intelectin shRNA. Both protein and mRNA levels of intelectin and nuclear factor-kappa B in brain tissue of mice were significantly decreased. Experimental findings suggest that Shuanghuanglian injection may downregulate nuclear factor-kappa B production via suppression of intelectin production, thus inhibiting inflammation associated with viral encephalitis.

[Effect of penetrative needling of scalp-acupoints on cerebral N-acetyl-aspartate and choline levels in intracerebral hemorrhage rabbits].

OBJECTIVE: To observe the effect of scalp-acupuncture on N-acetyl-aspartate (NAA) and choline (Cho) leveIs in the brain tissue in rabbits with intracerebral hemorrhage (ICH) so as to explore its underlying mechanisms in resisting neural lesion. METHODS: A total of 30 Newzealand rabbits were randomly divided into normal, model and acupuncture groups (n=10). ICH model was duplicated by injection of the rabbit-own blood (200 microL) into the bilateral Basal Nuclei (A: 2 mm; R, L; 5 mm; H: 10 mm). A filiform needle was inserted into "Baihui" (GV 20) to "Taiyang" (EX-HN 5) on the hemorrhage side, manipulated for 1 min, and then retained for 30 min (manipulated once again every 10 min). The treatment was conducted once daily for 7 days. Contents of NAA, Cho and creatine (Cr) in the intracerebral peri-hematoma tissues were detected by using 1H-Magnetic Resonance Spectroscopy (MRS). RESULTS: Compared with the normal group, NAA/Cr ratios at the time points of 2 h, 3 d and 7 d after ICH in the model group were decreased markedly (P < 0.05), while Cho/Cr ratios at the same 3 time-points after ICH in the model group were increased markedly (P < 0.05). In comparison with the model group, NAA/Cr ratio on day 3 after ICH in the acupuncture group was obviously higher (P < 0.05), while Cho/Cr ratios on day 3 and 7 were significantly down-regulated in the acupuncture group (P < 0.05). No significant changes were found in NAA/Cr ratios at 2 h and on day 7 and in Cho/Cr ratio at 2 h after ICH between the acupuncture group and the model group. CONCLUSION: Penetrative acupuncture of GV 20-EX-HN 5 is effective in suppressing ICH-induced increase of Cho/Cr ratio in ICH rabbits, which may contribute to its function in accelerating neural repair process.

[Sterols of marine bryozoan Bugula neritina from the South China Sea].

OBJECTIVE: To study the sterol constituents of Bugula neritina from the South China Sea. METHODS: The alcohol extract of Bugula neritina was purified by silica gel column and Sephadex LH-20 column chromatography, and structures of the isolated compounds were identified by spectroscopic analysis and comparison with those of literatures. RESULTS: Ten sterols were isolated and identified from the petroleum ether fraction of alcohol extract of B. neritina L.:Cholest-4-en-3-one(I); cholesterol(II);3beta,5alpha,9alpha-trihydroxy-(22E,24R)-ergosta-7,22-dien-6-one(III); 24-methyl-5alpha-cholesta-7,22-diene-3beta,5,6beta-triol(IV); 3beta-Hydroxy-7-methoxy-Cholesta-5-en(V);5alpha, 8alpha-epidioxy-(22E,24R)-ergosta-6, 22-dien-3beta-ol(VI); 3beta-hydroxycholest-5-en-7-one (VII); and 6beta-hydroxy-cholest-4-en-3-one(VIII); Cholesta-5-ene-3beta,7beta-diol(IX); Cholesta-5,22(E)-dien-3beta,7alpha-diol(X). CONCLUSION: Compounds IV-X were isolated from Bugula neritina for the first time.

[Effect of Shuanghuanglian injection on cerebral nuclear factor-kappaB expression in mice with viral encephalitis].

OBJECTIVE: To observe the effect of Shuanghuanglian injection on cerebral expression of nuclear factor-kappaB (NF-kappaB) in mice with viral encephalitis. METHODS: The mice with experimental viral encephalitis received treatment with Shuanghuanglian injection at the dose of 0.2, 1.5, and 5 for 5, 10 or 20 consecutive days. The total RNA of the brain tissue was extracted to analyze the protein and mRNA expression of NF-kappaB using Western blotting and RT-PCR, respectively. RESULTS: Compared with the control group, the mice with experimental viral encephalitis showed significantly increased protein and mRNA expressions of NF-kappaB (P<0.01). Treatment with Shuanghuanglian injection at the doses of 0.2 and 1.5 mg/kg significantly lowered NF-kappaB protein and mRNA expressions in the brain of mice with viral encephalitis (P<0.05), and the effect was even more obvious at the dose of 5 mg/kg (P<0.01). CONCLUSION: Shuanghuanglian injection can reduce the expression of NF-kappaB in the brain of mice with viral encephalitis in a dose- and time-dependent manner.