Transcriptomic Analysis for Diurnal Temperature Differences Reveals Gene-Regulation-Network Response to Accumulation of Bioactive Ingredients of Protocorm-like Bodies in Dendrobium officinale.

Dendrobium officinale Kimura et Migo (D. officinale) is one of the most important traditional Chinese medicinal herbs, celebrated for its abundant bioactive ingredients. This study demonstrated that the diurnal temperature difference (DIF) (T1: 13/13 °C, T2: 25/13 °C, and T3: 25/25 °C) was more favorable for high chlorophyll, increased polysaccharide, and total flavonoid contents compared to constant temperature treatments in D. officinale PLBs. The transcriptome analysis revealed 4251, 4404, and 4536 differentially expressed genes (DEGs) in three different comparisons (A: 25/13 °C vs. 13/13 °C, B: 13/13 °C vs. 25/25 °C, and C: 25/13 °C vs. 25/25 °C, respectively). The corresponding up-/down-regulated DEGs were 1562/2689, 2825/1579, and 2310/2226, respectively. GO and KEGG enrichment analyses of DEGs showed that the pathways of biosynthesis of secondary metabolites, carotenoid biosynthesis, and flavonoid biosynthesis were enriched in the top 20; further analysis of the sugar- and flavonol-metabolism pathways in D. officinale PLBs revealed that the DIF led to a differential gene expression in the enzymes linked to sugar metabolism, as well as to flavonol metabolism. Certain key metabolic genes related to ingredient accumulation were identified, including those involved in polysaccharide metabolism (SUS, SUT, HKL1, HGL, AMY1, and SS3) and flavonol (UGT73C and UGT73D) metabolism. Therefore, these findings indicated that these genes may play an important role in the regulatory network of the DIF in the functional metabolites of D. officinale PLBs. In a MapMan annotation of abiotic stress pathways, the DEGs with significant changes in their expression levels were mainly concentrated in the heat-stress pathways, including heat-shock proteins (HSPs) and heat-shock transcription factors (HSFs). In particular, the expression levels of HSP18.2, HSP70, and HSF1 were significantly increased under DIF treatment, which suggested that HSF1, HSP70 and HSP18.2 may respond to the DIF. In addition, they can be used as candidate genes to study the effect of the DIF on the PLBs of D. officinale. The results of our qPCR analysis are consistent with those of the transcriptome-expression analysis, indicating the reliability of the sequencing. The results of this study revealed the transcriptome mechanism of the DIF on the accumulation of the functional metabolic components of D. officinale. Furthermore, they also provide an important theoretical basis for improving the quality of D. officinale via the DIF in production.

Ferroptosis: a new mechanism of traditional Chinese medicine for cancer treatment.

Ferroptosis, distinct from apoptosis, is a novel cellular death pathway characterized by the build-up of lipid peroxidation and reactive oxygen species (ROS) derived from lipids within cells. Recent studies demonstrated the efficacy of ferroptosis inducers in targeting malignant cells, thereby establishing a promising avenue for combating cancer. Traditional Chinese medicine (TCM) has a long history of use and is widely used in cancer treatment. TCM takes a holistic approach, viewing the patient as a system and utilizing herbal formulas to address complex diseases such as cancer. Recent TCM studies have elucidated the molecular mechanisms underlying ferroptosis induction during cancer treatment. These studies have identified numerous plant metabolites and derivatives that target multiple pathways and molecular targets. TCM can induce ferroptosis in tumor cells through various regulatory mechanisms, such as amino acid, iron, and lipid metabolism pathways, which may provide novel therapeutic strategies for apoptosis-resistant cancer treatment. TCM also influence anticancer immunotherapy via ferroptosis. This review comprehensively elucidates the molecular mechanisms underlying ferroptosis, highlights the pivotal regulatory genes involved in orchestrating this process, evaluates the advancements made in TCM research pertaining to ferroptosis, and provides theoretical insights into the induction of ferroptosis in tumors using botanical drugs.

Clinical effect of stem cell transplantation combined with 308-nm excimer laser therapy for 56 cases of vitiligo.

OBJECTIVE: To probe the clinical effect of stem cell transplantation in combination with 308-nm excimer laser therapy for vitiligo and to analyze its value in clinical application. METHODS: A total of 56 patients with stable non-segmental vitiligo in different parts who were not cured by other therapies visiting our hospital from March 2019 to December 2021 were enrolled as study subjects. They were treated by stem cell transplantation combined with 308-nm excimer laser therapy. The treatment efficacy was observed and analyzed. RESULTS: Among the 56 patients, 38 (67.85%) and 49 (87.5%) patients were cured at 6 and 12 months after treatment, respectively. CONCLUSION: Stem cell transplantation combined with 308-nm excimer laser therapy for vitiligo achieves significant efficacy, with the cure rate far superior to that of other therapies for vitiligo. The therapy is worthy of popularization in the clinic.

Effects of intensive speech treatment on Mandarin speakers with Parkinson's Disease: A review.

BACKGROUND: Benefits of intensive speech treatment have been documented for a range of speech signs in English speakers with Parkinson's Disease (PD). However, the answer to a critical question that whether the same treatment benefits speech variables including intelligibility in Mandarin speakers is still unclear. In order to develop a targeted speech treatment for Mandarin speakers with PD, we reviewed the efficacy of intensive speech treatment to improve vocal loudness and functional communication and discuss possible explanations for efficacy on Mandarin speakers with PD. METHODS: Literatures about intensive speech treatment for Mandarin speakers with PD were retrieved from PubMed, Web of Science, Embase, China National Knowledge Infrastructure (CNKI), Wanfang and Weipu Database for Chinese Technical Periodicals (VIP) Database. Search strategy was (voice therapy OR speech therapy OR voice treatment OR speech treatment OR voice training OR speech training OR voice rehabilitation OR speech rehabilitation OR Lee Silverman voice treatment OR intensive speech treatment) and (Parkinson's disease) and (Mandarin speakers OR Chinese OR Chinese people). RESULTS: Five randomized controlled trials were selected and possible explanations for efficacy on individuals with PD are discussed. Further research directions are suggested. CONCLUSION: The existing evidence from treatment efficacy studies of intensive speech treatment provides support for improving vocal loudness, speech intelligibility, pitch and rate in Mandarin speakers with PD. Our future research will continue to work to conduct a large sample multicenter randomized controlled trial to provide high quality evidence and understand the basic mechanisms accompanying treatment-related change.

Safety and efficacy of wrist-ankle acupuncture in treating catheter-related bladder discomfort after transurethral resection of the prostate: a double-blind randomized clinical trial.

Background: Benign prostatic hyperplasia (BPH) is an age-related condition and its prevalence has increased as China's population ages. Transurethral resection of the prostate (TURP) remains the gold standard for treating moderate to severe BPH. Routine placement of a urinary catheter after TURP is often associated with catheter-related bladder discomfort (CRBD). The development of CRBD is related to an increased synthesis of prostaglandin (PG), and wrist-ankle acupuncture (WAA) can inhibit the expression of PG at the site of inflammation, thus alleviating CRBD symptoms. Here we evaluated the efficacy of WAA in alleviating CRBD in patients undergoing TURP. Methods: A total of 46 patients who underwent elective TURP in Hebei Provincial Hospital of Traditional Chinese Medicine from June 2022 to July 2022 were randomly divided into two groups according to the complete randomization method. The WAA group (n=23) and the control group (n=23). The WAA group received WAA, and the needles were retained for 24 h. The control group was treated with sham needles that did not penetrate the skin, and the needles were also retained for 24 h. At T1 (0 h after entering the ward), T2 (0.5 h after entering the ward), T3 (6 h after entering the ward), and T4 (24 h after entering the ward), CRBD severity score, visual analogue scale (VAS) and vital signs monitor were used for assessment. Accidents were recorded in the case report form. Graded data using Wlicoxon signed rank sum test, repeated measures using repeated measures analysis of variance. Results: A total of 46 patients participated in this study, and 44 patients completed the experiment. At T2, T3, and T4, the severity of CRBD in the WAA group was significantly lower than that in the control group (all P<0.05), and the VAS pain score was significantly lower in the WAA group than in the control group (all P<0.05). In contrast, the vital signs, including mean arterial pressure (MAP), heart rate (HR), and blood oxygen saturation, showed no statistical significance (all P>0.05). No accident occurred in both groups. Conclusions: WAA can effectively relieve CRBD symptoms after TURP. WAA deserves further research and assessment for clinical practice. Trial Registration: Chinese Clinical Trial Registry identifier: ChiCTR2200061525..

Platycodon grandiflorus polysaccharides deeply participate in the anti-chronic bronchitis effects of platycodon grandiflorus decoction, a representative of "the lung and intestine are related".

Platycodon grandiflorus (Jacq.) A. DC. (PG) root is one of the most commonly used medicine-food materials for respiratory discomfort in Asia, usually in the form of a decoction or leaching solution. As everyone knows, both of decoction and leaching solution is a polyphase dispersion system, containing low-molecular-weight water-soluble active ingredients and hydrophilic macromolecules. This study aimed to discuss the synergistic effect of Platycodon grandiflorus polysaccharide (PGP) and platycodin D (PD) in PG decoction against chronic bronchitis (CB) and the mechanism underlying. A series of PGP, PD, and PGD + PD suspensions were administrated to CB model rats, on the levels of whole animal and in situ intestinal segment with or without mesenteric lymphatic vessels ligation. It exhibited that PGP exhibited synergistic effects with PD, on improving the histopathological abnormity, mucus secretion excess, and immunological imbalance in lung of CB model rat, closely associated with its modulations on the mucosal immunity status in small intestine. The polysaccharide macromolecules in PG decoction or leaching solution should be responsible for the modulation of pulmonary immune state, possibly through the common mucosal immune between small intestine and lung. These results might be a new perspective that illustrates the classical theory of "the lung and intestine are related" in traditional Chinese medicine.

Coptis chinensis polysaccharides dynamically influence the paracellular absorption pathway in the small intestine by modulating the intestinal mucosal immunity microenvironment.

BACKGROUND: Traditional Chinese Medicine decoctions (TCMDs) can be used to prepare outstanding pharmaceutical preparations by the patient themselves. Small molecular active ingredients and macromolecular polysaccharides are inevitably co-existed in TCMDs. Different from the pharmacological synergies among small molecules, the macromolecular polysaccharides in TCMDs might contribute to disease treatment in several ways, although it is frequently overlooked. HYPOTHESIS/PURPOSE: This study proposes that the oral bioavailability of the water-insoluble alkaloids of Coptis chinensis Franch. (Ranunculaceae) (C. chinensis) decoction may be attributed to the co-existing C. chinensis polysaccharides (CCPs) dynamically influencing the small intestine microenvironment and regulating the modulation of the paracellular absorption pathway. METHODS: First, the effects of CCPs on the oral bioavailability of the main active ingredient of C. chinensis, berberine, were evaluated in vivo. Next, a series of in situ experimental models of intestinal perfusion and models of isolated jejunal mucosa, Caco-2 cell monolayer membranes, and microfold-like cells were established to assess the correlation among CCPs, intestinal mucosal immunity, and paracellular absorption in the small intestine. RESULTS: It was observed that CCPs could be endocytosed by the microfold cells on the surface of Peyer's patches, allowing CCPs to activate the lymphocytes, modulate the balance of Th1/Th2, control the secretion of immune effectors IFN-γ and IL-4, and finally regulate the tight junctions in the intestinal epithelial cells. This was a dynamic process with the movement of CCPs in the gastrointestinal tract that altered the flora distribution and functioning of the TLR/NF-κB signal pathway in the small intestine. CONCLUSION: The dynamical regulation of CCP on the immune microenvironment of small intestine is responsible for its promotion on the health controlling effects of C. chinensis in traditional dosage forms of decoction.

[CiteSpace knowledge mapping of traditional Chinese medicine in treatment of premature ovarian failure].

This study aimed to investigate the research trend of traditional Chinese medicine(TCM) against premature ovarian fai-lure(POF) from 1989 to 2021 by bibliometrics and explore the research status, research hotspots, and advances in international co-operation, knowledge structure, and active topics.The research articles on POF published from database inception to December 28, 2021, were retrieved from Web of Science and China National Knowledge Infrastructure(CNKI) and visually analyzed for countries, journals, authors, institutions, and keywords by CiteSpace 5.8.R3.A total of 1 468 articles were included, including 217 in English and 1 251 in Chinese.Since 1989, there has been an overall upward trend in the number of articles, with China serving as the main contributor.The core authors of Chinese articles are from a cooperative team represented by FENG Yi-xuan, REN Yu-lan, LING Le-le, and TENG Xiu-xiang.BETTERLE C is the author with the highest number of published articles in this international research field.The articles are mainly published by TCM journals and universities, and Human Reproduction accounts for the highest proportion of publications in the international research(11 articles, 5.07%).In the retrieved research articles, the research contents mainly focus on the treatment methods, research methods, and mechanism of action of TCM in the treatment of POF, where "Zuogui Pills" "gene" "cell" "model" "expression", etc.are the current research hotspots. "Acupuncture" "data mining" "systematic review" "oxidative stress" "activation" may be the potential topics in the follow-up research development.Future development should focus on the scientific interpretation and analysis of the theory and practice of TCM by modern scientific and technological methods.The research on informatization, digitization, and knowledge of TCM theory and practice is pivotal to promoting the internationalization and modernization of TCM, which can help researchers explore new directions for future research and identify new perspectives for potential collaboration in the field.

77Se-13C based dipolar correlation experiments to map selenium sites in microcrystalline proteins.

Sulfur-containing sites in proteins are of great importance for both protein structure and function, including enzymatic catalysis, signaling pathways, and recognition of ligands and protein partners. Selenium-77 is an NMR active spin-1/2 nucleus that shares many physiochemical properties with sulfur and can be readily introduced into proteins at sulfur sites without significant perturbations to the protein structure. The sulfur-containing amino acid methionine is commonly found at protein-protein or protein-ligand binding sites. Its selenium-containing counterpart, selenomethionine, has a broad chemical shift dispersion useful for NMR-based studies of complex systems. Methods such as (1H)-77Se-13C double cross polarization or {77Se}-13C REDOR could be valuable to map the local environment around selenium sites in proteins but have not been demonstrated to date. In this work, we explore these dipolar transfer mechanisms for structural characterization of the GB1 V39SeM variant of the model protein GB1 and demonstrate that 77Se-13C based correlations can be used to map the local environment around selenium sites in proteins. We have found that the general detection limit is ~ 5 Å, but longer range distances up to ~ 7 Å can be observed as well. This study establishes a framework for the future characterization of selenium sites at protein-protein or protein-ligand binding interfaces.

Clinical and pathological features and risk factors for primary breast cancer patients.

BACKGROUND: Breast cancer is the most common malignancy in women all around the world. According to the latest statistics in 2018, there were more than 2.08 million new breast cancer cases all around the world and more than 620000 deaths; the proportion of breast cancer deaths in women with cancer is 15%. By studying age, clinicopathological characteristics and molecular classification, age at menarche, age at birth, number of births, number of miscarriages, lactation time, surgical history of benign breast lesions, history of gynecological diseases, and other factors, we retrospectively summarized and compared the disease history of patients with primary breast cancer and patients with benign thyroid tumors admitted to our hospital in the past 10 years to explore the clinicopathological characteristics and risk factors for primary breast cancer. AIM: To investigate the clinical and pathological features and risk factors for primary breast cancer treated at our center in order to provide a reference for the prevention and treatment of breast cancer in the Zhuhai-Macao region. METHODS: Through a retrospective case-control study, 149 patients with primary breast cancer diagnosed and treated at Zhuhai Hospital of Guangdong Provincial Hospital of Traditional Chinese Medicine from January 2013 to March 2020 were included as a case group, and 165 patients with benign breast tumors diagnosed and treated from January 2019 to March 2020 were included as a control group. The data collected included age, age at menarche, age at first birth, number of births, number of miscarriages, lactation time, history of surgery for benign breast lesions, history of familial malignant tumors, history of gynecological diseases, history of thyroid diseases, and the tumor characteristics of the patients in the case group including pathological diagnosis, pathological type, tumor size, lymph node metastasis, distant metastasis, stage, and molecular classification, among others. In the case group, the chi-square test was used to analyze the clinical and pathological features of patients in three age groups (< 40, 40-59, and ≥ 60 years). A multifactor logistic regression analysis was used to analyze correlations between the two groups. RESULTS: Among 149 patients with primary breast cancer, the average age was 48.20 ± 12.06 years, and the proportion of patients at 40-59 years old was the highest, accounting for 61.8% of cases. The molecular type was mainly luminal B type, accounting for 69.2% of cases, and at the time of diagnosis, the tumor stage was mainly stage I/II, accounting for 62.4% of cases. There were no statistically significant differences in the distributions of tumor location, pathological type, tumor size, lymph node metastasis, stage, or molecular classification among the three age groups (< 40, 40-59, and ≥ 60 years) (P ≥ 0.05). The differences in the distribution of distant metastasis among the three age groups (< 40, 40-59, and ≥ 60 years) were statistically significant (P < 0.01). The differences in lactation time, history of familial malignant tumors, history of gynecological diseases, and history of thyroid diseases between the two groups were not statistically significant (P ≥ 0.05). The differences in age at disease diagnosis, age at menarche, and history of surgery for benign breast lesions were statistically significant (P < 0.01). The difference in age at first birth was also statistically significant (P < 0.05). CONCLUSION: The highest incidence of breast cancer in the Zhuhai-Macao region is present among women aged 40-59 years. There is a larger proportion of stage I/II patients, and the luminal B type is the most common molecular subtype. Distant metastasis occurs mainly in the ≥ 60-year-old group at the first diagnosis; increased age, late age at menarche, and late age at first birth may be risk factors for primary breast cancer, and a history of surgery for benign breast lesions may be a protective factor for primary breast cancer.

[Effect of Dendrobium officinale superfine powder on stomach Yin deficiency model mice induced by "spicy overeating"].

Dendrobium officinale is a traditional Chinese medicine for nourishing Yin and benefiting stomach. Its superfine powder has many advantages, such as good dissolution, high utilization rate, strong integrity and easy to use. However, the researches on effect of D. officinale superfine powder on stomach Yin deficiency model are still not sufficient. In this experiment, we explored the effect of D. officinale superfine powder in mice model with stomach Yin deficiency caused by "spicy overeating", and provided certain reference value for its application in gastrointestinal diseases. Male ICR mice were randomly divided into normal group, model group, Yiweitang group, omeprazole group, and D. officinale superfine powder high, medium and low dose groups. The mixture of wine and pepper liquid was given by gavage administration for 30 d, and the corresponding drug was given for 60 d while the model was conti-nued. The body weight, food intake, water intake, fecal moisture content and particle number, foot temperature of mice were measured. The levels of serum gastrin(Gas), motilin(MTL) and somatostatin(SS) were measured by ELISA. Gastric histomorpho-logy was observed by HE staining. The expression levels of nuclear factor kappa B(NF-κB) and cyclooxygenase-2(COX-2) were determined by immunohistochemistry. The expression levels of B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) in gastric tissues were detected by Western blot. The results showed that D. officinale superfine powder could increase the food intake, water intake, fecal moisture content and particle number, reduce the foot temperature, improve the pathological changes of gastric mucosa, reduce the expression of NF-κB, COX-2 protein in gastric tissues, and increase the ratio of Bax/Bcl-2. D. officinale superfine powder can "nourish Yin and benefit the stomach", improve the syndrome of stomach Yin deficiency, such as "hunger but not want to eat, dry mouth but not want to drink, hand and feet hot, constipation", and reduce the damage of gastric mucosa. The mechanism may be related to regulating the secretion of gastrointestinal hormones, inhibiting the inflammation of gastric tissues and promoting the apoptosis of abnormal cells in gastric tissues.

[Effect of Dendrobium officinale on Yin deficiency constipation induced by warm-drying medicine in mice].

Dendrobium officinale is a sacred product for nourishing Yin and has a clear "thick gastrointestinal" effect. Modern pharmacological studies had found that it could improve gastrointestinal function. This study observed the improvement effect of D. officinale on constipation model mice with Yin deficiency caused by warm-drying medicine. It provided experimental basis for the treatment of Yin deficiency constipation. The male and female ICR mice were randomly divided into normal group, model group, D. officinale high, medium and low dose groups(0.6, 0.4, 0.2 g·kg~(-1)), and phenolphthalein tablets group. The model mice of Yin deficiency constipation were established by gavage with warm-drying medicine. The overall state and body temperature of the mice were observed and recorded. The number of feces, feces weight, fecal moisture content and intestinal propulsion were measured. The morphological damage of colon tissue was observed by hematoxylin-eosin(HE) staining. The expression of inducible nitric oxide synthase(iNOS) in the colon was detected by Western blot and immunohistochemical method. The expression of iNOS mRNA in the colon was detected by Real-time fluorescence quantitative PCR, and the serum cyclic guanosine phosphate(cGMP) level was detected the enzyme-linked immunosorbent assay(ELISA). The results showed that D. candidum could reduce the body temperature of mice with Yin deficiency constipation, increase the number of feces, wet feces, dry feces and intestinal propulsion ability, reduce the expression of iNOS protein and mRNA in the colon, and reduce the content of cGMP in the serum. It showed that D. candidum could improve the symptoms of Yin deficiency constipation mice caused by warm-drying medicine, and the mechanism may be related to reducing the expression of iNOS in the colon and increasing intestinal motility.

Self-assembled aggregations in Coptidis Rhizoma decoction dynamically regulate intestinal tissue permeability through Peyer's patch-associated immunity.

Objective: To investigate the dynamic regulation of self-assembled aggregations (SAA) in Coptidis Rhizoma decoction on the permeability of intestinal tissue and the mechanism underlying. Methods: The effects of SAA on berberine (Ber) absorption were respectively analyzed in an in situ intestinal perfusion model and in an Ussing Chamber jejunum model with or without Peyer's patches (PPs). The expression levels of ZO-1, Occludin and Claudin-1 were detected by immunofluorescence to evaluate the tight junction (TJ) between intestinal epithelium cells. The expression levels of T-box-containing protein expressed in T cells, signal transducers and activators of tranion-6, retinoic acid receptor-related orphan receptor γt and forkhead box P3 in PPs were detected by the reverse transcription-polymerase chain reaction and the secretions of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-ß (TGF-ß) in PPs were evaluated by immunohistochemistry, to reflect the differentiation of T lymphocyte in PPs to helper T (Th) cell 1, Th2, Th17 and regulatory T (Treg) cell. To confirm the correlation between SAA in Coptidis Rhizoma decoction, PPs-associated immunity and intestinal epithelium permeability, SAA were administrated on an Ussing Chamber jejunum model with immunosuppressed PPs and evaluated its influences on intestinal tissue permeability and TJ proteins expression. Results: SAA in Coptidis Rhizoma decoction could dose-dependently promote Ber absorption in jejunum segment, with the participation of PPs. The dose-dependent and dynamical regulations of SAA on permeability of intestinal tissue and TJ proteins expression level between intestinal epithelium cells occurred along with the dynamically changed T lymphocyte differentiation and immune effectors secretion in PPs. The administration of SAA on immunosuppressed PPs exhibited dose-dependent PPs activation, inducing dynamic promotion on intestinal tissue permeability and inhibition on TJ proteins expression. Conclusion: SAA can improve the Ber absorption in small intestine, through the PPs-associated immunity induced dynamic regulation on intestinal tissue permeability and TJ proteins expression. These findings might enlighten the research of traditional Chinese medicine decoction.

Coptis chinensis Franch polysaccharides provide a dynamically regulation on intestinal microenvironment, based on the intestinal flora and mucosal immunity.

ETHNOPHARMACOLOGICAL RELEVANCE: Coptis chinensis Franch is one of the most widely used traditional Chinese herbs in China and was firstly recorded in "Shennong's Classic of Materia Medica" in the Han Dynasty. The medical records in past thousands years have fully confirmed the clinical efficacies of Coptis chinensis Franch against intestinal diseases. The polysaccharides in herbal medicines can be digested by the flora and uptaken by the Peyer's patches (PPs) in intestine. It can be reasonably presumed that the polysaccharides in Coptis chinensis Franch (CCP) should be one of the critical element in the regulation of intestinal microenvironment. AIM OF THE STUDY: This study intended to explore the dynamic regulation of CCP on intestinal microenvironment from the perspective of the intestinal mucosal immunity and the intestinal flora, in order to provide a new research perspective for the pharmacological mechanism of Coptis chinensis Franch. MATERIALS AND METHODS: The absorption and distribution of CCP in intestinal tissues were observed after the perfusion of FITC labeled CCP. The influences of CCP on intestinal flora were evaluated by the 16sRNA gene illumina-miseq sequencing after gavage. The regulations of CCP on intestinal mucosal immunity were evaluated by the immunohistochemical analysis of the interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-ß (TGF-ß) secretion in PPs and intestinal epithelial tissue. RESULTS: With the self-aggregation into particles morphology, CCP can be up-taken by PPs and promote the IFN-γ, IL-4, IL-17 and TGF-ß secretion in PPs in a dose-dependent manner. The CCP can also be utilized by the intestinal flora and dynamically regulate the diversity, composition and distribution of the intestinal flora. The temporal regulations of CCP on IFN-γ, IL-4, IL-17 and TGF-ß secretions in intestinal epithelial tissues are consistent with the variation tendency of intestinal flora. CONCLUSION: CCP can provide effective, dynamical and dose-dependent regulations on intestinal microenvironment, not only the intestinal flora but also the PPs and intestinal epithelium related immune response. These may be involved in the multiple biological activities of Coptis chinensis Franch.

Role of particle aggregates in herbal medicine decoction showing they are not useless: considering Coptis chinensis decoction as an example.

A large number of plants and fungi are homologs of medicine and food, and are used in the form of decoctions for complementary foods, dietary cures, and disease therapy in traditional medicine. Besides the widespread concern around the physiological and pharmacological actions of the active ingredients, the phase change in decoction and its influences on the active ingredients' absorption should not be ignored. Lots of particle aggregates are generated during the decoction of herbal medicine and then end up being taken together with the active ingredients. The question arises, "Is the absorption of active ingredients associated with the particle aggregates in decoction?" The present study takes the Coptis chinensis decoction (CCD), the particle aggregates in CCD (CCD-Ps), and the water-insoluble active ingredient of Berberine (Ber) as typical examples to investigate the effects of particle aggregates in herbal medicine decoction on the active ingredient absorption in the intestine and the underlying mechanisms. The CCD-Ps are mainly composed of polysaccharide, with commonly features of a hundred-nanometers size and negatively charged. A series of Coptis chinensis polysaccharide (CCP) composed particle aggregates (CCP-Ps) were self-assembled to mimic the CCD-Ps. In situ single-pass intestinal perfusion experiments exhibited that, both the CCD-Ps and CCP-Ps exhibited charge-dependent promotion on Ber absorption in the intestine, through regulating the tight junctions (TJs) between intestinal epithelia cells. Caco-2 cell monolayer model experiments revealed that the particle aggregates not only promoted paracellular Ber transport through TJs regulation but also improved the transcellular Ber transport through active transport and endocytosis. The present study provides a novel viewpoint to explain the scientific implications of herbal medicine decoction, in which the particles aggregated in decoction are not useless but rather act as an effective and important enhancer for adsorption of the active ingredients through multiple mechanisms.

Imperative and effective reversion of synovial hyperplasia and cartilage destruction in rheumatoid arthritis through multiple synergistic effects of O2 and Ca2.

Abnormal synovial hyperplasia and cartilage destruction in a joint cavity are the key causes affecting the pain and disability in rheumatoid arthritis (RA) and, unfortunately, there exists no effective treatment for them. This investigation reports an effective reversion of the above pathological characteristics in RA owing to the use of a prolonged O2/Ca2+-supporting phototherapy hydrogel. The performed in vitro and in vivo experiments exhibit that the prolonged O2-supporting not only promotes the direct cell-killing effects of singlet oxygen, but also persistently blocks the pathological feedback between the abnormal proliferation of fibroblast-like synoviocyte and the local oxygen depletion. Furthermore, the Ca2+, which is the other decomposition product of the O2 donor, induces mitochondrial Ca2+ overload and endoplasmic reticulum Ca2+ disorder and triggers Ca2+-associated apoptosis and immunogenic cell death. In addition to these multiple synergistic effects on synovial hyperplasia, the prolonged Ca2+ support can also induce the regeneration of cartilage in RA affected joints. The present study may thus provide an effective therapeutic strategy for the prevention and reversion of joint lesions and the accompanying arthralgia and deformity in RA.

77Se NMR Probes the Protein Environment of Selenomethionine.

Sulfur is critical for the correct structure and proper function of proteins. Yet, lacking a sensitive enough isotope, nuclear magnetic resonance (NMR) experiments are unable to deliver for sulfur in proteins the usual wealth of chemical, dynamic, and structural information. This limitation can be circumvented by substituting sulfur with selenium, which has similar physicochemical properties and minimal impact on protein structures but possesses an NMR compatible isotope (77Se). Here we exploit the sensitivity of 77Se NMR to the nucleus' chemical milieu and use selenomethionine as a probe for its proteinaceous environment. However, such selenium NMR spectra of proteins currently resist a reliable interpretation because systematic connections between variations of system variables and changes in 77Se NMR parameters are still lacking. To start narrowing this knowledge gap, we report here on a biological 77Se magnetic resonance data bank based on a systematically designed library of GB1 variants in which a single selenomethionine was introduced at different locations within the protein. We recorded the resulting isotropic 77Se chemical shifts and relaxation times for six GB1 variants by solution-state 77Se NMR. For four of the GB1 variants we were also able to determine the chemical shift anisotropy tensor of SeM by solid-state 77Se NMR. To enable interpretation of the NMR data, the structures of five of the GB1 variants were solved by X-ray crystallography to a resolution of 1.2 Å, allowing us to unambiguously determine the conformation of the selenomethionine. Finally, we combine our solution- and solid-state NMR data with the structural information to arrive at general insights regarding the execution and interpretation of 77Se NMR experiments that exploit selenomethionine to probe proteins.

[Effect of Yunkang Oral Liquid on preventing LPS-induced abortion and regulating immune tolerance in mice].

To investigate the effect of Yunkang Oral Liquid on preventing lipopolysaccharide(LPS)-induced abortion and regulating immune tolerance in mice,sixty normal ICR mice were randomly divided into normal group,model group,Yunkang Oral Liquid high,middle and low dose groups and progesterone group.Abortion model was established by tail vain injection of LPS(0.1µg/mouse)on the 7th day of pregnancy.Since the first day of pregnancy,the same volume of distilled water,Yunkang Oral Liquid at the dose of 36,18 and 9 m L·kg~(-1)·d~(-1),and progesterone at the dose of 0.038 g·kg~(-1)·d~(-1)were given in corresponding groups.The mice were sacrificed at the 9th day of pregnancy,and the embryo loss of each group was calculated.The levels of Th1 type cytokines(TNF-α,IFN-γ)and Th2 type cytokines(IL-4,IL-10)in uterus homogenate were detected by ELISA.HE staining was performed to examine the histopathological changes in the decidua.The expression levels of CD14,F4/80 in macrophages of uterus were detected by immunohistochemistry.Western blot was used to investigate the protein expression of TLR4,MyD88 and NF-κB in uterine decidua.In our study,all Yunkang Oral Liquid groups could significantly reduce the embryo absorption rate of mice,while high dose group can significantly increase the levels of IL-10 and IL-4;both medium and high dose groups can significantly decrease TNF-α,and IFN-γlevelsin the uterus of model mice,reduce the protein expression of NF-κB,MyD88 and TLR4 in uterine decidua tissue.Various treatment groups could reduce the counts of F4/80,CD14 macrophages and decrease expression area in uterine tissue.Our results showed that Yunkang Oral Liquid could prevent LPS-induced abortion,and the mechanism may be associated with inhibiting the TLR4/MyD88/NF-κB signaling pathway and regulating the balance of Th1/Th2 immune factors,which could improve the endometrial receptivity of mice,and promote the development of decidua and implantation of embryo.

Growth performance, innate immune responses and disease resistance of fingerling blunt snout bream, Megalobrama amblycephala adapted to different berberine-dietary feeding modes.

A 8-week feeding trial was conducted to evaluate the effect of different berberine-dietary feeding modes on growth, non-specific immune responses and disease resistance of blunt snout bream, Megalobrama amblycephala. Fish (average initial weight 4.70 ± 0.02 g) were fed two fat levels (5% and 10%) diets in three berberine-feeding modes (supplementing 50 mg/kg berberine continuously, two-week or four-week intervals) with four replicates, respectively. Then, fish were challenged by Aeromonas hydrophila and mortality was recorded for the next 96 h after feeding trial. The results showed that different feeding modes of berberine significantly influenced growth, innate immunity and antioxidant capability of fish. Fish fed normal diet with 50 mg/kg berberine at two-week interval mode reflected remarkably (P < 0.05) high weight gain (WG). Plasma TC and TG contents were significantly (P < 0.05) decreased. The lysozyme (LYZ) activities, complement component 3 (C3) and complement component 4 (C4) concentrations were significantly (P < 0.05) increased. Fish not only exhibited relatively low hepatopancreas malondialdehyde (MDA) and lipid peroxide (LPO) contents, but also significantly (P < 0.05) improved superoxide dismutase (SOD) and catalase (CAT) activities. Fish mortality after challenged by Aeromonas hydrophila was decreased. Same results were also presented in fish fed high-fat diet with 50 mg/kg berberine at two-week, four-week intervals or continuous feeding modes. Based on fish healthy improvement and feeding cost saving, blunt snout bream fed normal diet with 50 mg/kg berberine at two-week interval or fed high-fat diet with berberine at two-week or four-week intervals were optimal feeding mode, respectively.

Structural characterization and antioxidant activities of polysaccharides extracted from the pulp of Elaeagnus angustifolia L.

In this study, two polysaccharides (Elaeagnus angustifolia L. polysaccharide-1 (PEA-1) and PEA-2) were prepared from Elaeagnus angustifolia L. Then, the preliminary structure and antioxidant activities of all the samples were investigated. The results showed that the average molecular weights for PEA-1 and PEA-2 were 9113 and 5020 Da, respectively. And, PEA-1 was mainly composed of rhamnose, xylose, mannose, glucose, and galactose, respectively. The components of PEA-2 were rhamnose, mannose, glucose, and galactose, respectively. Moreover, the Antioxidant assays demonstrated that PEA-1 possessed of strong free radicals scavenging activity and hydroxyl radicals scavenging activities, suggesting that PEA-1 could potentially be used as natural antioxidant.