Biochemical analysis reveals the systematic response of motion sickness mice to ginger (Zingiber officinale) extract's amelioration effect.

ETHNOPHARMACOLOGICAL RELEVANCE: As a common medicinal and edible plant, Zingiber officinale Roscoe (ginger) is often used for the prevention of motion sickness. However, the mechanism of its anti-motion sickness remains to be elucidated. AIM OF THE STUDY: To explore novel treatment for motion sickness with less side effects, anti-motion sickness effect of ginger (Zingiber officinale) extract (GE) and the possible molecular mechanisms were investigated. MATERIALS AND METHODS: The anti-motion sickness effect of ginger was evaluated through mice animal experimental models. Components of ginger that might contribute to the anti-motion sickness effect were analyzed by LC-MS/MS. Subsequently, biochemical analysis integrated with serum metabolomic profiling were performed to reveal the systematic response of motion sickness mice to ginger extract's amelioration effect. RESULTS: Exhaustive swimming time of mice in the GE group reached 8.9 min, which was 52.2% longer than that in the model group. Motion sickness index scores and time taken traversing balance beam of mice in the GE group were decreased by 53.2% and 38.5%, respectively. LC-MS/MS analysis suggested that various active ingredients in GE, such as gingerol, ginger oil and terpenoids, might contribute to its appealing anti-motion sickness activity. Biochemical analysis revealed that GE can relieve motion sickness through reducing histamine and acetylcholine release in vestibular system, regulating fatty acid oxidation, sugar metabolism and bile acid metabolism in mice. CONCLUSION: Gavage of mice with GE can effectively relieve the symptoms of autonomic nervous system dysfunction, improve the balance and coordination ability and ameliorate the ability to complete complex work after rotation stimulation. GE has attractive potential for development and utilization as novel anti-motion sickness food or drugs.

Protective effect of coix seed seedling extract on 60 Co-γ radiation-induced oxidative stress in mice.

Exposure to ionizing radiation (IR) can cause oxidative damage to human body, leading to various diseases and even death. In this study, the potential radioprotective effect of coix seed seedling extract (CSS-E) was studied through a model of 60 Co-γ radiation-induced oxidative stress in mice. Overall radioprotective effect of CSS-E against radiation-induced damage was evaluated by biochemical analysis and histopathological analysis. The results showed that CSS-E could significantly reduce the IR-induced damage to the hematopoietic system. CSS-E-M (200 mg/kg BW) pretreatment could increase the activities of superoxide dismutase in serum, liver, and spleen increased by 31.68%, 45.10%, and 56.67%, respectively, and the glutathione peroxidase levels in serum, liver, and spleen of mice were improved by 19.17%, 41.97%, and 130.56%, respectively. Meanwhile, the glutathione levels of serum, liver, and spleen in CSS-E-M group were increased by 17.10%, 35.06%, and 40.71%, respectively. The contents of MDA in different tissues and serum could be reduced by CSS-E-M treatment to the normal level. Moreover, CSS-E could markedly reduce the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in radiation mice, among which CSS-E-M group showed maximum restoration with decreased AST and ALT levels by 20.13% and 32.76% as compared against IR group. In conclusion, these results indicated that CSS-E could be used as a potential natural radioprotectant against IR-induced damage.

Wound-healing activity of glycoproteins from white jade snail (Achatina fulica) on experimentally burned mice.

Burns are a global public health problem and the treatment of burn wounds is a major medical and economic issue. White jade snails (Achatina fulica) are now widely distributed in Asia, and they have been used to treat burns in folk medicine of China. In this study, the glycoproteins from white jade snails were investigated and their effect on burn healing was evaluated by a mouse burn model. The results showed that the snail mucus was mainly composed of proteins and polysaccharides, and it had good adhesion. The main component of snail mucus was glycoprotein from the results of DEAE Sepharose FF ion exchange chromatography. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging effect of 1 mg/mL snail mucus reached 13.77%. The wound healing rate of the snail mucus group was higher than that of the control group (p < 0.0001). Histopathological results showed that mice in the snail mucus group had a faster healing than that of the control group. The biochemical analysis was in agreement with the histopathological findings. These results suggested that glycoproteins from snail mucus showed effective wound healing activities in the skin of experimentally burned mice.

Anti-fatigue activity of hemp leaves water extract and the related biochemical changes in mice.

To explore novel sources of anti-fatigue drugs and food, the anti-fatigue activity of hemp leaves water extract (HLWE) was investigated through exhaustive swimming tests of mice. The median exhaustion swimming time of mice gavaged with HLWE reached 55.4 min, which was 156.8% and 87.8% longer than that of the control group and Rhodiola group, respectively. Then, several biochemical parameters related to fatigue were determined to explore the possible anti-fatigue reasons. The blood lactic acid concentration of mice in HLWE group was 0.76 mmol/L, which was 24.8% lower than that in the control group. Compared with the control group, the glutathione peroxidases activity of mice in HLWE group increased by 296.2%. Based on the results, HLWE exhibited outstanding anti-fatigue activity through reducing the accumulation of lactic acid and improving the activities of defense antioxidant enzymes. It shows appealing potential for development and utilization as novel anti-fatigue food or drugs.

Comparative study on the bioactive components and in vitro biological activities of three green seedlings.

To explore functional food ingredients from green seedlings, the bioactive components (phenolic compounds and γ-aminobutyric acid) and antioxidant activities (DPPH radical scavenging ability, ABTS radical scavenging ability and reducing power) of three green seedlings, including coix seed seedling (CSS), highland barely seedling (HBS) and naked oats seedling (NOS) cultivars were respectively measured and deeply compared. Results indicated that CSS showed the highest contents of the total polyphenol (183.35 mg/100 g), total flavonoid (348.68 mg/100 g), and γ-aminobutyric acid (54.17 mg/100 g). As expected, CSS also exerted the highest level of antioxidant activity, followed by HBS and NOS. Moreover, CSS possessed the potential of stimulating immune responses, including promoting proliferation and strengthening phagocytosis function of RAW264.7 cells. Taken together, all results suggested that the three green seedlings, especially CSS could be used as natural ingredients for functional food.

LGR4 overexpression is associated with clinical parameters and poor prognosis of serous ovarian cancer.

OBJECTIVE: LGR4 expression in serous ovarian cancer paraffin-embedded tissues and fresh tissues were investigated, and its expression associated with clinicopathological parameters and prognosis in serous ovarian cancer was explored. METHODS: From Dec, 2009 to Jan, 2020, 122 paraffin-embedded serous ovarian cancer patients and 41 paired paratumor tissues who were both diagnosed and operated at the memorial hospital of Sun Yat-sen University and Integrated Hospital of Traditional Chinese Medicine, Southern Medical University were selected in this research, respectively, and all of these tissues were performed by immunohistochemistry (IHC) with a polyclonal antibody for LGR4. Meanwhile, from Aug, 2013 to Mar, 2019, 15 cases of serous ovarian cancer fresh tissues and 15 cases of paratumor fresh tissues who were operated at Integrated Hospital of Traditional Chinese Medicine, Southern Medical University were performed with Quantitative Real-time PCR to detect the mRNA expression of LGR4, respectively. RESULTS: LGR4 expression was much higher both in paraffin-embedded and fresh cancer tissues than that in paratumor tissues, respectively, and its expression was associated with recurrence free survival and overall survival in serous ovarian cancer patients. Moreover, in a multivariate model LGR4 was an indeed independent predictor of poor survival in serous ovarian cancer patients. CONCLUSION: LGR4 is upregulated in serous ovarian cancer, and LGR4 is an indeed useful independent prognostic predictor in serous ovarian cancer, and it may provide important clinical value of serous ovarian cancer.

[Establishment of new evaluation standards for systemic anaphylactoid reactions using mouse model].

The detection of drug-induced anaphylactoid reactions remains a global challenge,still lacking mature and reliable animal models or test methods. Therefore,the purpose of this paper is to explore and establish the test methods and evaluation standards for anaphylactoid reactions that apply to injection drugs. Based on the anaphylactoid reaction symptoms of mice induced by intravenous injection drugs C48/40 and Tween 80,a list of systemic anaphylactoid reaction symptoms in mice was sorted out and an evaluation standard of anaphylactoid reactions symptoms was established by applying symptom intensity coefficient K( that can represent these verity of anaphylactoid reaction symptoms) and its calculation formula Accordingly,histamine,tryptase,and Ig E were selected as blood indicators of anaphylactoid reactions,so that a test method combining symptoms evaluation and blood makers detection was established.This test method could be used to evaluate the characteristics of anaphylactoid reactions: coefficient K,blood histamine levels were highly and positively correlated with C48/80 and Tween 80 dose; The log value of histamine was highly and positively correlated with K; tryptase level may rise,or remain steady,or drop,possibly associated with the characteristics of the tested object and time for blood taking; and Ig E level would drop or remain steady,but it would not rise,which can be clearly distinguished from type I allergic reactions. On this basis,tiohexol,iopromide,paclitaxel,Xuesaitong Injection,Shuanghuanglian Injection and Shengmai Injection were used to investigate the applicability. The testing results showed a high degree of consistency with the actual clinical situation. The results suggest that the method of systemic anaphylaxis test in mice has high sensitivity,specificity and good consistency with clinical practice.It is suggested to be further validated and popularized.

Anti-Disuse Osteoporosis Activity of a Complex of Calcium-Binding Peptide from Auricularia auricula Protein Hydrolysates.

Osteoporosis is a common metabolic bone disease that is often seen in bedridden patients and astronauts. Long-term bed rest and nonweight bearing tend to induce disuse osteoporosis. Calcium supplements are commonly used to help treat disuse osteoporosis along with medications, most of which are calcium carbonate based, but they have poor absorption effects. In this study, we prepared a novel Auricularia auricula peptide-calcium complex (AP-Ca) and evaluated its protective effects on disuse osteoporosis. In vitro assays showed that AP-Ca significantly increased the contents of calcium (P < 0.05) and the activity of alkaline phosphatase (AKP; P < 0.05) of osteoblasts cultured in a two-dimensional-rotating wall vessel. Meanwhile, supplementation with AP-Ca also inhibited the production of pro-inflammatory factors induced by the loss of stress, especially TNF-α (P < 0.05). In vivo, a mouse tail suspension (TS) model was established, and the results showed that AP-Ca helped to improve bone mineral density, bone mineral content, and bone organic content in TS mice and effectively alleviated the alteration of enzymes related to bone metabolism, including AKP (P < 0.05) and serum tartrate-resistant acid phosphatase (P < 0.05), to avoid more serious bone loss induced by TS. Furthermore, we found that AP-Ca downregulated the bone resorption-associated pro-inflammatory genes interleukin-1 (IL-1), tumor necrosis factor-α, and IL-6 by 59.53 ± 3.55%, 48.01 ± 5.68%, and 40.00 ± 5.89%, respectively (P < 0.05). In conclusion, AP-Ca showed potential to suppress bone loss induced by disuse and might be considered a new alternative to reduce the risk of disuse osteoporosis. PRACTICAL APPLICATION: This peptide-calcium complex supplement exhibited protective effects on the bone loss induced by disuse, which provided a new alternative for patients and astronauts to reduce the risk of disuse osteoporosis.

Extraction and characterization of pectin from Premna microphylla Turcz leaves.

In this work, response surface methodology (RSM) and microwave pretreatment were used to extract pectin from Premna microphylla Turcz leaves (PMTL). The process variables were optimized by the isovariant central composite design to improve the pectin extraction yield. The optimum conditions obtained were as follows, extraction time 2 h, temperature 90 °C, pH 2 and liquid-solid (LS) ratio 50 mL/g. The extraction yield was 18.25% under these conditions, which was close to the predicted value (17.60%). Then the pectin was characterized by gas chromatographic (GC), spectrophotometric (UV-visible Spectroscopy) and spectroscopic (Fourier transform infrared) methods. The galacturonic acid content was 82.75%, and on this basis, the other monosaccharide composition analysis illustrated that pectin from PMTL was also composed of rhamnose, arabinose, mannose, glucose and galactose in a ratio of 2.96:1.17:1.04:8.07:2.05. The pectin of PMTL had an esterification degree of 62.50% and it showed good antioxidant activity. Taken together, the pectin of PMTL could be used as potential additive in food industry.

Anti-tumor properties of anthocyanins from Lonicera caerulea 'Beilei' fruit on human hepatocellular carcinoma: In vitro and in vivo study.

In this study, the anthocyanin from Lonicera caerulea 'Beilei' fruit (ABL) was extracted and purified. The purified component (ABL-2) was then evaluated for its anti-tumor properties on human hepatoma cells (SMMC-7721) in vitro and the murine hepatoma cells (H22) in vivo. In vitro, ABL-2 not only significantly inhibited the growth of SMMC-7721 cells, but also remarkably blocked the cells' cycle in G2/M phase, inducing DNA damage and eventually leading to apoptosis. In vivo, ABL also killed tumor cells, inhibited tumor growth, and improved the survival status of H22 tumor-bearing mice. These effects were associated with an increase in the activities of antioxidase and a decrease in the level of lipid peroxidation, as evidenced by changes in SOD, GSH-Px, GSH, and MDA levels. In addition, ABL-2 also regulated the levels of immune cytokines including IL-2, IFN-γ, and TNF-α. These results revealed that ABL-2 exerts an effective anti-tumor effect by dynamically adjusting the REDOX balance and improving the immunoregulatory activity of H22 tumor-bearing mice. High performance liquid chromatography (HPLC) analysis revealed that cyanidin-3,5-diglucoside (8.16 mg/g), cyanidin-3-glucoside (387.60 mg/g), cyanidin-3-rutinoside (23.62 mg/g), and peonidin-3-glucoside (22.20 mg/g) were the main components in ABL-2, which may contribute to its anti-tumor activity.

Protective mechanisms of purified polyphenols from pinecones of Pinus koraiensis on spleen tissues in tumor-bearing S180 mice in vivo.

The previous study evaluated the antitumor activity and the underlying mechanism of the purified polyphenols from pinecones of Pinus koraiensis (PPP-40) using a tumor-bearing S180 mice model. This study was designed to evaluate the protective effects of PPP-40 on spleen tissues of S180 mice in vivo. Pretreatment with PPP-40 (150 mg per kg BW per D) could significantly inhibit tumor growth, enhance spleen index and prevent the decline of haematological parameters of S180 mice induced by the tumor microenvironment. Moreover, the treatment with PPP-40 was shown to significantly inhibit splenocyte apoptosis by TUNEL staining and flow cytometry, characterized by the inhibition of splenocyte cycle (G0/G1) arrest, increase in the percentages of splenic T lymphocytes (CD3+ T cells) and T cell subsets (CD3+CD4+ and CD3+CD8+ T cells), as well as the production of T cell-related cytokines (IL-2, IL-12, and TNF-α) in splenocytes exposed to the tumor microenvironment. These effects were associated with a decrease in oxidative stress, as evidenced by the changes in the SOD, GSH-Px, GSH and MDA levels of liver and spleen tissues of S180 mice. Furthermore, the protective effect of PPP-40 on spleen tissues was deeply analyzed by detecting apoptosis-related proteins using immunohistochemistry staining. The results indicated that the protective multi-mechanisms of action also were associated with the inhibition of apoptosis through down-regulation protein expressions of Bax, caspase-9, caspase-8 caspase-3, Fas and up-regulation of the expressions of Bcl-2. These results suggested that PPP-40 is a natural antitumor agent and possesses strong immunomodulatory activities by protecting the spleen tissues of tumor-bearing S180 mice.

Optimization of Purification, Identification and Evaluation of the in Vitro Antitumor Activity of Polyphenols from Pinus Koraiensis Pinecones.

In this study, an efficient purification method for the polyphenols of Pinus koraiensis pinecone (PPP) has been developed. AB-8 resin was verified to offer good adsorption and desorption ratio for PPP. Response surface methodology (RSM) indicated that the optimized purification parameters for PPP were 1.70 mg GAE/mL phenolic sample concentration, 22.00 mL sample volume, and 63.00% ethanol concentration. Under these conditions, the experimental purity of PPP was 27.93 ± 0.14% (n = 3), which matched well with the predicted purity of 28.17%. Next, the antiproliferative effects of PPP on seven cancer cell lines, including A375 (human skin melanoma cancer cell line), A549 (human lung cancer cell line), SH-SY5Y (human neuroblastoma cell line), LOVO (human colon cancer stem cell line), MCF-7 (human breast cancer cell line), HeLa (human cervical cancer line), and HT29 (human colon cancer line), were examined by MTT assays. The results indicated that PPP had the highest capacity for inhibiting LOVO cells growth with an EC50 value of 0.317 ± 0.0476 mg/mL. Finally, Ultra-high performance liquid chromatography- tandem mass spectrometry (UPLC-MS) was used to tentatively identify twenty-four peaks in the purified PPP, of which five representative peaks were identified as catechin, methyl quercetin, o-vanillin, luteolin and coronaric acid. Our results demonstrate that Pinus koraiensis pinecone is a readily available source of polyphenols, and the purified PPP could be a promising natural antitumor agent for applications in functional foods.

[Evaluation criteria for rat skin anaphylactoid test].

This study is to explore characteristic indexes in evaluation criteria for rat skin anaphylactoid test comparing skin blue spot OD values at the treated position and the control position in the same animal. Common contrast agents, traditional Chinese medicine injections and injections' active pharmaceutical ingredients or excipients in the existing clinical anaphylactoid reaction reports were taken as test drugs in the rat skin anaphylactoid test to define the K value: K > 2 represents positive anaphylactoid reaction, 1.2 ≤ K ≤ 2 represent doubtable anaphylactoid; K < 1.2 represents negative anaphylactoid reaction, which were taken as the criteria for evaluating anaphylactoid of tested drugs. The evaluation result and that for classic criteria were compared to study the applicability of K value. According to the comparison, K value, as the evaluation criteria in the rat skin anaphylactoid test, can more truly reflect the actual situation of skin aizen and minimize the error caused by animal individual factors. Compared with positive and negative two-level criteria for blue spot diameter, K value's positive, doubtable and negative three-level criteria are more objective and accurate. Therefore, K value can be used as the evaluation criteria in the rat skin anaphylactoid test.