Endophytic fungus Penicillium steckii DF33 promoted tanshinones biosynthesis in Salvia miltiorrhiza by regulating the expression of CYP450 genes.

Salvia miltiorrhiza, a prominent traditional Chinese medicinal resource, has been extensively employed in the management of cardiovascular and cerebrovascular ailments. Ensuring the consistency of S. miltiorrhiza raw materials revolves around the imperative task of maintaining stable tanshinones content and composition. An effective approach in this regard involves the utilization of endophytic fungi as inducers. Within this context, our study spotlights an endophytic fungus, Penicillium steckii DF33, isolated from the roots of S. miltiorrhiza. Remarkably, this fungus has demonstrated a significant capacity to boost the biosynthesis and accumulation of tanshinones. The primary objective of this investigation is to elucidate the underlying regulatory mechanism by which DF33 enhances and regulates the biosynthesis and accumulation of tanshinones. This is achieved through its influence on the differential expression of crucial CYP450 genes within the S. miltiorrhiza hairy roots system. The results revealed that the DF33 elicitor not only promotes the growth of hairy roots but also enhances the accumulation of tanshinones. Notably, the content of cryptotanshinone was reached 1.6452 ± 0.0925 mg g-1, a fourfold increase compared to the control group. Our qRT-PCR results further demonstrate that the DF33 elicitor significantly up-regulates the expression of most key enzyme genes (GGPPS, CPS1, KSL1, CYP76AH1, CYP76AH3, CYP76AK1, CYP71D411) involved in the tanshinone biosynthesis pathway. This effect is particularly pronounced in certain critical CYP450 genes and Tanshinone ⅡA synthase (SmTⅡAS), with their expression levels peaking at 7 days or 14 days, respectively. In summary, endophytic P. steckii DF33 primarily enhances tanshinone biosynthesis by elevating the expression levels of pivotal enzyme genes associated with the modification and transformation stages within the tanshinone biosynthesis pathway. These findings underscore the potential of employing plant probiotics, specifically endophytic and root-associated microbes, to facilitate the biosynthesis and transformation of vital constituents in medicinal plants, and this approach holds promise for enhancing the quality of traditional Chinese medicinal materials.

Aucubin alleviates methotrexate-induced enteritis in rats by inducing autophagy.

One of the toxic side effects of methotrexate (MTX) is enteritis. Aucubin, an iridoid glycoside derived from traditional medicinal herbs, has been proven to have anti-inflammation, anti-apoptosis and anti-oxidation properties. This work explored the effect and mechanism of aucubin in treating MTX-induced enteritis in a rat model. Two doses of aucubin (5 and 10 mg/kg) were adopted for the assessment of its pharmacological activity. We observed that in rats with MTX-induced enteritis, the body weight and small intestinal weight decreased. The intestine barrier was injured, as reflected by pathological examinations and an increase in D-lactate and diamine oxidase concentration in serum. Intestinal inflammation was shown by the observation of macrophages in the intestine and the concentrations of inflammatory cytokines tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum. The NLR family pyrin domain containing 3 (NLRP3) inflammasome was shown to be activated by the enhancement of NLRP3, cleaved-caspase 1, IL-18 and IL-1ß. Moreover, autophagy was reflected by transmission electron microscopy as slightly induced, along with changes in autophagy-related markers microtubule-associated protein 1 light chain 3 (LC3) and Beclin1. Remarkably, aucubin treatment attenuated the MTX-induced disease activity index increase, intestinal damage, inflammatory response and NLRP3 inflammasome activation, but provoked autophagy. Rapamycin, an autophagy activator, showed similar therapeutic effects to aucubin on MTX-induced enteritis. However, 3-methyladenine, an autophagy inhibitor, reversed the protective effects of aucubin. These findings prompted the hypothesis that aucubin alleviates MTX-induced enteritis by aggravating autophagy. This study might provide evidence for further investigation on the therapeutic role of aucubin in MTX-resulted enteritis.

Effect of Limosilactobacillus reuteri ZJF036 on Growth Performance and Gut Microbiota in Juvenile Beagle Dogs.

This experiment investigated the effects of Limosilactobacillus reuteri ZJF036 on growth performance, serum biochemical parameters, and gut microbiota in beagle dogs. Sixteen 75 ± 5-day-old healthy male beagles (4.51 ± 1.37 kg) were randomly divided into two groups; the experimental group (L1) and the control group (L0), and then fed with or without a basal diet containing L. reuteri ZJF036 (109 CFU/g), respectively. The results showed that there was no significant difference in daily weight gain between the two groups (P > 0.05). However, we found that L. reuteri ZJF036 decreased Chao1 index and ACE index and increased the relative abundance of Firmicutes and Fusobacteria (P < 0.05) compared to the L0 group. In addition, we also found that the ratio of Firmicutes to Bacteroidetes was decreased in L1 group. Furthermore, the relative abundance of Lactobacillus increased, while that of Turicibacter and Blautia decreased in L1 group (P < 0.05). In conclusion, L. reuteri ZJF036 appeared to regulate the intestinal microbiota of beagle dogs. This study revealed the potential use of L. reuteri ZJBF036 as a probiotic supplement for beagle dogs.

Causal association of genetically determined circulating vitamin D metabolites and calcium with multiple sclerosis in participants of European descent.

BACKGROUND: Vitamin D is an important regulator of calcium. Mendelian randomization (MR) studies exclusively focused on the circulating total 25-hydroxyvitamin D (25(OH)D) as a biomarker of vitamin D status, and have found the causal association between 25(OH)D and the risk of multiple sclerosis (MS). However, it currently remains unclear about the causal association of the 25(OH)D subtypes including 25(OH)D3 and C3-epi-25(OH)D3, as well as calcium with the risk of MS. METHODS: We performed a two-sample MR study to evaluate the causal association of circulating total 25(OH)D, 25(OH)D3, C3-epi-25(OH)D3, and calcium with the risk of MS using large-scale genome-wide association studies (GWAS) datasets from total 25(OH)D (n = 417,580), 25(OH)D3 (n = 40,562), C3-epi-25(OH)D3 (n = 40,562), calcium (n = 305,349), and MS (14,802 MS and 26,703 controls). We selected five MR methods including inverse-variance weighted (IVW), simple median, weighted median, MR-Egger, MR-PRESSO (Mendelian Randomization Pleiotropy Residual Sum and Outlier), and contamination mixture method. RESULTS: IVW showed that the genetically increased circulating 25(OH)D level (OR = 0.81, 95% CI: 0.70-0.94, P = 4.00E-03), circulating 25(OH)D3 level (OR = 0.85, 95% CI: 0.76-0.95, P = 5.00E-03), and circulating C3-epi-25(OH)D3 level (OR = 0.85, 95% CI: 0.74-0.98, P = 2.30E-02) were causally associated with reduced risk of MS. However, IVW showed no causal association between circulating calcium level and the risk of MS with OR = 2.85, 95% CI: 0.42-19.53, P = 2.85E-01. CONCLUSIONS: Our current findings together with evidence from other MR studies support the use of vitamin D but not calcium supplementation for the prevention of MS.

Low-frequency electroacupuncture improves disordered hepatic energy metabolism in insulin-resistant Zucker diabetic fatty rats via the AMPK/mTORC1/p70S6K signaling pathway.

BACKGROUND AND AIM: Disordered hepatic energy metabolism is found in obese rats with insulin resistance (IR). There are insufficient experimental studies of electroacupuncture (EA) for IR and type 2 diabetes mellitus (T2DM). The aim of this study was to probe the effect of EA on disordered hepatic energy metabolism and the adenosine monophosphate (AMP)-activated protein kinase (AMPK)/mammalian target of rapamycin complex 1 (mTORC1)/ribosomal protein S6 kinase, 70-kDa (p70S6K) signaling pathway. METHODS: Zucker Diabetic Fatty (ZDF) rats were randomly divided into three groups: EA group receiving EA treatment; Pi group receiving pioglitazone gavage; and ZF group remaining untreated (n = 8 per group). Inbred non-insulin-resistant Zucker lean rats formed an (untreated) healthy control group (ZL, n = 8). Fasting plasma glucose (FPG), fasting insulin (FINS), C-peptide, C-reactive protein (CRP) and homeostatic model assessment of insulin resistance (HOMA-IR) indices were measured. Hematoxylin-eosin (H&E) staining was used to investigate the liver morphologically. The mitochondrial structure of hepatocytes was observed by transmission electron microscopy (TEM). Western blotting was adopted to determine protein expression of insulin receptor substrate 1 (IRS-1), mTOR, mTORC1, AMPK, tuberous sclerosis 2 (TSC2) and p70S6K, and their phosphorylation. RT-PCR was used to quantify IRS-1, mTOR, mTORC1, AMPK and p70S6K mRNA levels. RESULTS: Compared with the ZF group, FPG, FINS, C-peptide, CRP and HOMA-IR levels were significantly reduced in the EA group (p < 0.05, p < 0.01). Evaluation of histopathology showed improvement in liver appearances following EA. Phosphorylation levels of AMPK, mTOR and TSC2 decreased, and IRS-1 and p70S6K increased, in hepatocytes of the ZF group, while these negative effects appeared to be alleviated by EA. CONCLUSIONS: EA can effectively ameliorate IR and regulate energy metabolism in the ZDF rat model. AMPK/mTORC1/p70S6K and related molecules may represent a potential mechanism of action underlying these effects.

Forsythiaside A alleviates methotrexate-induced intestinal mucositis in rats by modulating the NLRP3 signaling pathways.

Most chemotherapeutic drugs can kill the tumor cells, but also cause a vast damage to body, such as intestinal mucositis (IM). The present study was design to find out the effect of Forsythiaside A (FTA) on chemotherapeutic-induced IM in rats. Briefly, for 3 consecutive days, male Sprague-Dawley rats were treated with 7 mg / kg methotrexate (MTX) to establish IM and simultaneously administered with 40 or 80 mg / kg FTA for 7 days. Our results showed that the final body weight and daily food intake were increased, and the disease activity index was reduced in the MTX group after FTA treatment. The MTX group showed the pathological alterations like the inflammatory cells infiltration, the mucosal layer destruction, glands expansion, intestinal villi structure disorder and goblet cells reduction, while we found that 80 mg / kg FTA treatment displayed evident reversal effects. ELISA further suggested that TNF-α, IL-1ß and IL-18 levels in serum in MTX-induced rats were reduced after 80 mg / kg FTA treatment. Moreover, FTA decreased the number of leukocytes, neutrophils and lymphocytes in peripheral blood. Western blot and immunofluorescence results indicated that the expression levels of NLRP3, cleaved caspase 1, cleaved IL-1ß and CD68 positive rate were down-regulated in MTX-induced rats after 80 mg / kg FTA intervention. The findings of the current study suggested that FTA effectively inhibited MTX-induced IM in rats by attenuating the activation of the NLRP3 signaling pathways.

Endophytic fungus Cladosporium tenuissimum DF11, an efficient inducer of tanshinone biosynthesis in Salvia miltiorrhiza roots.

Salvia miltiorrhiza is a traditional medicinal plant mainly used for cardiovascular and cerebrovascular disease treatment. Tanshinones are the main bioactive constituents of S. miltiorrhiza, which mainly accumulate around its root periderm tissue. Endophytic fungi are important bioelicitors or probiotics that can promote the accumulation of secondary metabolites and sustainable cultivation of medicinal plants. Among them, endophytic Cladosporium spp., possessing a variety of biotransformation and metabolic abilities, is an ideal elicitor source. Here, we used a gnotobiotic system to investigate the effects of the endophytic fungus Cladosporium tenuissimum DF11 on tanshinone biosynthesis in S. miltiorrhiza roots. The results showed that C. tenuissimum DF11 mainly colonizes the intercellular space of the root tissues and promotes tanshinone biosynthesis and accumulation in S. miltiorrhiza roots by upregulating the expression of the genes encoding for key enzymes HMGR, DXS, DXR, GGPPS, CPS, KSL and CYP76AH1 of the tanshinone biosynthesis pathway. The expression levels of almost all genes encoding for key enzymes reached the response peak in the first or second week after DF11 colonization. Taken together, the endophytic fungus C. tenuissimum DF11 could promote secondary metabolite accumulation in S. miltiorrhiza roots. These results indicate that DF11 will be a potential biofertilizer fungus to regulate and stabilize the quality of cultivated S. miltiorrhiza medicinal materials.

Effect of Vitamin A Supplementation on Growth Performance, Serum Biochemical Parameters, Intestinal Immunity Response and Gut Microbiota in American Mink (Neovison vison).

This experiment investigated the effect of vitamin A supplementation on growth, serum biochemical parameters, jejunum morphology and the microbial community in male growing-furring mink. Thirty healthy male mink were randomly assigned to three treatment groups, with 10 mink per group. Each mink was housed in an individual cage. The mink in the three groups were fed diets supplemented with vitamin A acetate at dosages of 0 (CON), 20,000 (LVitA) and 1,280,000 IU/kg (HVitA) of basal diet. A 7-day pretest period preceded a formal test period of 45 days. The results show that 20,000 IU/kg vitamin A increased the ADG, serum T-AOC and GSH-Px activities, villus height and villus height/crypt depth ratio (p < 0.05). The mRNA expression levels of IL-22, Occludin and ZO-1 in the jejunum of mink were significantly higher in the LVitA group than those in the CON and HVitA groups (p < 0.05). Vitamin A supplementation increased the diversity of jejunum bacteria, decreased the ratio of Firmicutes to Bacteroidetes and increased the relative abundance of Akkermansia, uncultured bacterium f Muribaculaceae, Allobaculum, Lachnospiraceae NK4A136 group, Rummeliibacillus and Parasutterella. The comparison of potential functions also showed enrichment of glycan biosynthesis and metabolism, transport and catabolism pathways in the vitamin A supplementation groups compared with the CON group. In conclusion, these results indicate that dietary vitamin A supplementation could mediate host growth by improving intestinal development, immunity and the relative abundance of the intestinal microbiota.

Effect of plasma vitamin C levels on Parkinson's disease and age at onset: a Mendelian randomization study.

BACKGROUND: Until now, epidemiological evidence regarding the association between vitamin C intake (both diet and supplements) and Parkinson's disease (PD) remains inconsistent. Hence, it is necessary to establish the causal link between vitamin C levels and PD, and further develop effective therapies or prevention. METHODS: We selected 11 newly identified plasma vitamin C genetic variants from a large-scale plasma vitamin C GWAS dataset (n = 52,018) as the effective instrumental variables, and extracted their corresponding GWAS summary statistics from PD (33,674 PD cases and 449,056 controls) and PD age at onset (AAO) (n = 28,568). We then performed a Mendelian randomization (MR) study to evaluate the causal association of plasma vitamin C levels with PD and PD AAO using inverse-variance weighted (IVW), the weighted median, MR-Egger, and MR-PRESSO test. RESULTS: We did not observe any significant association between genetically increased vitamin C levels and PD. Interestingly, we found a reduced trend of PD AAO (1.134 years) with 1 SD genetically increased vitamin C levels using IVW (beta = - 1.134, 95% CI: [- 2.515, 0.248], P = 0.108). Importantly, this trend was further successfully verified using both weighted median and MR-Egger. Each 1 SD genetically increased vitamin C levels could reduce PD AAO 1.75 and 2.592 years using weighted median (beta = - 1.750, 95% CI: [- 3.396, - 0.105], P = 0.037) and MR-Egger (beta = - 2.592, 95% CI: [- 4.623, - 0.560], P = 0.012). CONCLUSIONS: We demonstrated the causal association between genetically increased plasma vitamin C levels and reduced PD AAO in people of European descent. Randomized controlled trials are required to clarify whether diet intake or supplement, or both could reduce the AAO of PD.

Endophytic fungus Mucor circinelloides DF20 promote tanshinone biosynthesis and accumulation in Salvia miltiorrhiza root.

As a traditional Chinese medicine, Salvia miltiorrhiza rhizome is mainly used to treat cardiovascular diseases. Symbiosis of endophytic fungi with their host plants, is an effectively regulatory means to promote the growth and secondary metabolism of medicinal plants. Here, an endophytic fungus Mucor circinelloides DF20 was co-cultivated with the sterile seedlings of S. miltiorrhiza, to clarify the promoting mechanism on tanshinone biosynthesis and accumulation in S. miltiorrhiza root. The assay of promoting-growth activities in vitro showed that DF20 have the ability to produce IAA and siderophores. DF20 could significantly promote the biosynthesis and accumulation of tanshinones in the root of S. miltiorrhiza, especially the content of tanshinone ⅡA, reaching 4.630 ± 0.342 mg/g after 56 days of DF20 treatment, which is 22-fold of the control group. The result also showed that the hyphae of M. circunelloides DF20 mainly colonized in the root tissue interspace of S. miltiorrhiza, and a small amount of hyphae were located inside the cells. The results of florescent real-time quantitative RT-PCR showed that DF20 colonization significantly increase the expression level of some key enzyme genes (DXS, DXR, HMGR, GGPPS) in tanshinone biosynthesis pathway, but the regulatory effect mainly occurred in the early stage of co-culture, while the expression level decreased in different degrees in the later stage. In conclusion, the endophytic fungus M. circunelloides DF20 can form an interaction relationship with its host, then to promote the biosynthesis and accumulation of tanshinones in root by upregulating the key enzyme genes expression levels of the biosynthesis pathway.

Impact of Serum Calcium Levels on Alzheimer's Disease: A Mendelian Randomization Study.

BACKGROUND: Altered calcium homeostasis is hypothesized to underlie Alzheimer's disease (AD). However, it remains unclear whether serum calcium levels are genetically associated with AD risk. OBJECTIVE: To develop effective therapies, we should establish the causal link between serum calcium levels and AD. METHODS: Here, we performed a Mendelian randomization study to investigate the causal association of increased serum calcium levels with AD risk using the genetic variants from a large-scale serum calcium genome-wide association study (GWAS) dataset (61,079 individuals of European descent) and a large-scale AD GWAS dataset (54,162 individuals including 17,008 AD cases and 37,154 controls of European descent). Here, we selected the inverse-variance weighted (IVW) as the main analysis method. Meanwhile, we selected other three sensitivity analysis methods to examine the robustness of the IVW estimate. RESULTS: IVW analysis showed that the increased serum calcium level (per 1 standard deviation (SD) increase 0.5 mg/dL) was significantly associated with a reduced AD risk (OR = 0.57, 95% CI 0.35-0.95, p = 0.031). Meanwhile, all the estimates from other sensitivity analysis methods were consistent with the IVW estimate in terms of direction and magnitude. CONCLUSION: In summary, we provided evidence that increased serum calcium levels could reduce the risk of AD. Meanwhile, randomized controlled study should be conducted to clarify whether diet calcium intake or calcium supplement, or both could reduce the risk of AD.

[Promoting tanshinone synthesis of Salvia miltiorrhiza root by a seed endophytic fungus, Phoma herbarum D603].

The interaction of endophytes and host plant is an effective mean to regulate the growth and secondary metabolism of medicinal plants. Here we want to elucidate the effects and mechanism of Phoma herbarum D603 on the root development and tanshinone synthesis in root of Salvia miltiorrhiza by endophyte-plant coculture system. The mycelium of P. herbarum D603 was colonized in the root tissue space, and formed a stable symbiotic relationship with host plant. The in vitro activities analysis showed that the concentration of IAA produced by D603 can reach(6.45±0.23) µg·mL~(-1), and this strain had some abilities of phosphorus solubilization and siderophore production activities. The coculture experiment showed that strain D603 can significantly promote the synthesis and accumulation of tanshinones in the root of S. miltiorrhiza, in which after 8 weeks of treatment with D603, the content of tanshinone Ⅱ_A in the roots reached up to(1.42±0.59) mg·g~(-1). By the qRT-PCR analysis results, we found that D603 could improve the expression levels of some key genes(DXR, DXS, GGPP, HMGR, CPS) of tanshinone biosynthesis pathway in host plant S. miltiorrhiza, but the promoting effect mainly occurred in the early stage of the interaction, and the enzyme activity level decreased in varying degrees of the later stage. In summary, seed-associated endophyte P. herbarum D603 can promote the growth and root development of S. miltiorrhiza by producing hormones, promoting nutrient absorption and siderophore production, and promote the synthesis and accumulation of tanshinones by regulating the expression level of key genes in the synthetic pathway in S. miltiorrhiza.

Changes in Intestinal Microbiota of Type 2 Diabetes in Mice in Response to Dietary Supplementation With Instant Tea or Matcha.

OBJECTIVE: Gut microbiota plays a key role in metabolism and health in diabetes patients with gastrointestinal microbiota dysbiosis. Thus, regulating the ecological balance of gut microbiota may provide a pathway toward improvement for these patients. Our previous study showed that functional ingredients in tea may inhibit cornstarch digestion in vitro. METHODS: A cornstarch-tea diet was developed, and in this study we investigated the effects of such a diet on blood glucose and gut microbiota in diabetic mice. RESULTS: Diabetes resulted in significant weight loss, hyperphagia and hyperglycemia. 16S rDNA sequencing revealed that in diabetes there is significantly increased Bacteroidaceae, Helicobacteraceae, Ruminococcaceae, Enterobacteriaceae, Rikenellaceae and Saccharibacteria_genera_incertae_sedis, and significantly decreased Lactobacillaceae, Prevotellaceae, Coriobacteriaceae, Verrucomicrobiaceae and Bifidobacteriaceae. The cornstarch‒tea diet resulted in a trend toward reduced blood glucose, with particularly increased levels of Coriobacteriaceae, Lactobacillaceae, Prevotellaceae and Bifidobacteriaceae, and decreased Bacteroidaceae, Ruminococcaceae, Helicobacteraceae and Enterobacteriaceae. CONCLUSIONS: Instant tea and matcha supplementation had beneficial effects on regulation of blood glucose and gut microbiota, reversing the changes in microbiota caused by alloxan injection. The cornstarch‒tea regulation pathway is involved in bacterium group regulation rather than single-species regulation, which suggests that cornstarch combined with tea may be used as a functional food supplement for diabetes patients.

Progressive study of the effect of superfine green tea, soluble tea, and tea polyphenols on the physico-chemical and structural properties of wheat gluten in noodle system.

In this study, the improving effects of green tea powder, soluble tea, and tea polyphenols on the mixing and tensile qualities of dough and texture of tea-enriched noodles, as well as the physico-chemical and structural properties of gluten proteins were progressively investigated. Dough strength and noodle texture were significantly increased by all the three tea products. Tea polyphenols in particular presented the most effective improvement with highest dough stability, resistance, and noodle chewiness. SEM indicated that tea products all induced a more developed gluten network, and polyphenol noodle showed the most continuous and ordered structure. FT-IR and fluorescence spectrum indicated that tea polyphenols promoted an enhancement in α-helix structure and the hydrophobic interactions. Tea polyphenols induced the SH/SS interchange during processing and cooking, and enhanced the water-solids interaction in noodles. AFM results showed that polyphenols induced the polymerization of gluten protein molecular chains, with increased chain height and width.

Circulating Vitamin D Levels and Alzheimer's Disease: A Mendelian Randomization Study in the IGAP and UK Biobank.

Observational studies strongly supported the association of low levels of circulating 25-hydroxyvitamin D (25OHD) and cognitive impairment or dementia in aging populations. However, randomized controlled trials have not shown clear evidence that vitamin D supplementation could improve cognitive outcomes. In fact, some studies reported the association between vitamin D and cognitive impairment based on individuals aged 60 years and over. However, it is still unclear that whether vitamin D levels are causally associated with Alzheimer's disease (AD) risk in individuals aged 60 years and over. Here, we performed a Mendelian randomization (MR) study to investigate the causal association between vitamin D levels and AD using a large-scale vitamin D genome-wide association study (GWAS) dataset and two large-scale AD GWAS datasets from the IGAP and UK Biobank with individuals aged 60 years and over. Our results showed that genetically increased 25OHD levels were significantly associated with reduced AD risk in individuals aged 60 years and over. Hence, our findings in combination with previous literature indicate that maintaining adequate vitamin D status in older people especially aged 60 years and over, may contribute to slow down cognitive decline and forestall AD. Long-term randomized controlled trials are required to test whether vitamin D supplementation may prevent AD in older people especially those aged 60 years and may be recommended as preventive agents.

[Clinical therapeutic effect of hyperlipidemia of turbid phlegm obstruction pattern/syndrome treated with the different Jin's three-needle therapies].

OBJECTIVE: To compare the differences in clinical effects on hyperlipidemia of turbid phlegm obstruction pattern/syndrome treated with the different Jin's three-needle therapies so as to provide a new approach and theoretic evidence for the clinical optimal scheme of acupuncture on hyperlipidemia. METHODS: A total of 90 patients were randomly divided into education group, electroacupuncture group and catgut embedding group (30 cases in each). The routine health education was given to the education group. On the base of the treatment as the education group, electroacupuncture was applied to the electroacupuncture group for 30 min each time, twice a week; and the catgut embedding was applied to the catgut embedding group at the same acupoints as the electroacupuncture group, once a week. The treatment was given consecutively for 8 weeks in each group. Before and after treatment, the obesity indices [weight, waistline, body mass index (BMI)], blood lipid indices[serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C)] and insulin resistance indices [fasting blood glucose(FBG), fasting plasma insulin(FINS), homeostasis model assessment-estimated insulin resistance (HOMA-IR)] were observed in the three groups before and after treatment. RESULTS: After treatment, in the electroacupuncture group and the catgut embedding group, the results of the obesity indices, e.g. weight, waistline and BMI, the levels of blood lipid, e.g.TC, TG, LDL-C and the insulin resistance indices, e.g. FINS and HOMA-IR were all obviously decreased compared with those before treatment (P<0.05, P<0.01), and the level of HDL-C apparently increased than that before treatment (P<0.01). In the catgut embedding group, FBG was obviously decreased after treatment as compared with that before treatment (P<0.01). After treatment, the improvements in LDL-C and HDL-C in the catgut embedding group were superior to those in the electroacupuncture group (P<0.05). Regarding the obesity indices and insulin resistance indices, the differences were not statistically significant in comparison between the catgut embedding group and the electroacupuncture group (all P>0.05). The total effective rate was 90.0% in the catgut embedding group (27/30) and it was 83.3% (25/30) in the electroacupuncture group, either of them was better than 60.0% (18/30) in the education group , indicating the statistical significance (P<0.01, P<0.05). CONCLUSION: On the base of health education, either the catgut embedding therapy or electroacupuncture of Jin's three-needle treatment achieves the positive regulation on the abnormal lipid metabolism and insulin resistance. The hypoglycemic effect and the impro-ving effect in LDL-C and HDL-C of the catgut embedding therapy are superior to electroacupuncture.

[Progress of researches on mechanisms of acupuncture therapy for insulin resistance].

Insulin resistance (IR) is a common pathophysiological basis of many chronic diseases, such as obesity, type 2 diabetes mellitus (T2DM), nonalcoholic fatty liver disease, cerebral vascular disease, cardiovascular and neurodegenerative diseases, etc. Acupuncture therapy has been demonstrated to have a positive role in reducing IR level in clinical practice. In the present paper, we summarized development of researches on the mechanism of acupuncture therapy underlying improvement of IR in recent 10 years from 1) regulating expression of hypothalamic phosphatidylinositol-3-kinase (PI3K)-p85 and PI3K-P110 proteins in obesity rats; 2) regulating the levels of some related proteins of insulin target tissues (liver and skeletal muscle), such as insulin receptor substrate-1 (IRS)-1 and -2, glucose transporter mRNAs and proteins, sterol regulatory element-binding protein 1 (SREBP-1) and fatty acid synthetase proteins; 3) suppressing inflammatory reaction of liver tissue and down-regulating serum adiponectin in T2DM rats; 4) raising the activity of superoxide dismutase (SOD) and reducing the levels of reactive oxygen species, and malondialdehyde (MDA) contents of quadriceps femoris in spontaneous IR rats; and 5) attenuating structural injury of pancreas islet and apoptosis of pancreatic ß cells in diabetes rats. Multi-levels and various systems of the neuro-endocrine-immune networks are involved in the actions of acupuncture in the improvement of IR. In the future, more attention should be paid to the study on the acupoint specificity, suitable acupoint combinations and stimulus parameters in clinical treatment of IR related various metabolic diseases, further optimizing clinical treatment protocols.

Effect of dietary fibers on the structure and digestibility of fried potato starch: A comparison of pullulan and pectin.

The granular morphology, long-range and short-range ordered structures of fried potato starch were measured in the absence and presence of the dietary fibers. The in vitro digestibility of the fried starchy samples was also quantified using the Englyst method with logarithm-of-slope (LOS) analysis. After frying, the starch granules disintegrated, their internal crystalline structure disappeared, and the quantity of double helices present decreased. As a result of these changes, the fried starch was digested rapidly. Addition of pullulan or pectin to the samples prior to frying, reduced the structural changes observed in the starch granules during frying. Consequently, the fractions of slowly digestible and resistant starch (SDS and RS) increased significantly in the presence of the dietary fibers. These effects were attributed to the ability of the dietary fibers to sequester some of the water, thereby reducing starch granule structural changes, as well as due to their ability to coat the starch granules and interfere with the starch digestion process.

Impact of amylose content on structural changes and oil absorption of fried maize starches.

The absorption of oil during frying has important implications for food quality, cost, and nutrition. Maize starches with low (WMS), intermediate (NMS), and high amylose (HAMS) contents were therefore heated in oil to mimic the frying process, and the impact of amylose content on the hierarchical structures and oil absorption of the fried starches was evaluated. Amylose affected the oil absorption by interfering with the structural evolution of the starch or by directly interacting with the lipids during frying. At low moisture level (20%), the granular state was preserved after frying and so the size and porosity of the granules played a dominant role in the oil absorption process, explaining why the highest oil absorption occurred in WMS. At 40% moisture content, NMS absorbed the most oil because of its granular morphology and lower crystallinity. At 60% moisture content, HAMS absorbed more oil than NMS, because more amylose molecules in HAMS provided more hydrophobic helical cavities available for lipids.

[Eletroacupuncture improves lipid metabolic disorder by regulating hepatic AMPK/p38 MAPK/RRARγ signaling in rats with high-fat diet-induced insulin resistance].

OBJECTIVE: To observe the effect of eletroacupuncture (EA) intervention on lipid metabolism and expression of AMP-activated kinase (AMPK), p38 mitogen-activated protein kinase (p38 MAPK) and peroxisome proliferator-activated receptor γ (PPARγ) protein in the liver in rats with insulin resistance (IR), so as to reveal its mechanisms underlying improvement of IR. METHODS: Forty male SD rats were randomly divided into blank control, model, medication, and EA groups (n＝8 in each). The IR model was established by feeding the rats with high-fat diet for 12 weeks. After successful establishment of model, the rats in the blank control group and model group were fixed in the self-made rat bag without receiving any treatment. The rats in the medication group were treated by gavage of pioglitazone (10 mL/kg). EA (2 Hz /100 Hz, 1 mA) was applied to bilateral "Fenglong"(ST40) and "Sanyinjiao"(SP6) for 20 min, once a day, for continuous 14 days for rats in the EA group. The ultrastructure of the liver tissue was observed by transmission electron microscope (TEM). Blood samples were taken from the abdominal aorta for detecting serum C-peptide (C-P), adiponectin (ADP), leptin (LEP) and resistin (RES) contents using enzyme linked immunosorbent assay (ELISA). The expression levels of AMPK, p38 MAPK and PPARγ proteins in the liver tissue were detected by Western blot. RESULTS: After modeling, the contents of serum C-P, LEP and RES, and the expression of liver p38 MAPK protein were significantly up-regulated (P<0.01, P<0.05), and the content of ADP and expression of AMPK and PPARγ significantly down-regulated in the model group compared with the blank control group (P<0.01). The increased contents of C-P, LEP and RES, and p38 MAPK protein expression and the decreased serum ADP and hepatic AMPK and PPARγ expression levels were completely reversed in both the EA and medication groups relevant to the model group (P<0.01, P<0.05). No significant differences were found between the EA and medication groups in up-regulating the levels of ADP, AMPK and PPARγ and in down-regulating the levels of C-P, LEP, RES and p38 MAPK(P>0.05). Outcomes of TEM showed that morphological structure of liver mitochondria was damaged, including a large number of lipid droplets, being blur in appearance, rupture of partial membrane, dissapearance of partial mitochondrial crests with vacuolus-like appearance and decrease of rough endoplasmic reticulum in the model group, which was relatively milder in both EA and medication groups. CONCLUSION: EA intervention is able to improve the disorder of lipid metabolism of IR rats, which may be associated with its effects in lowering the activity of fatty acid synthesis-related enzymes and regulating AMPK/p38 MAPK/PPARγ signaling to improve IR in the liver tissue.