Inhibition of DNA demethylation enhances plant tolerance to cadmium toxicity by improving iron nutrition.

Although the alteration of DNA methylation due to abiotic stresses, such as exposure to the toxic metal cadmium (Cd), has been often observed in plants, little is known about whether such epigenetic changes are linked to the ability of plants to adapt to stress. Herein, we report a close linkage between DNA methylation and the adaptational responses in Arabidopsis plants under Cd stress. Exposure to Cd significantly inhibited the expression of three DNA demethylase genes ROS1/DML2/DML3 (RDD) and elevated DNA methylation at the genome-wide level in Col-0 roots. Furthermore, the profile of DNA methylation in Cd-exposed Col-0 roots was similar to that in the roots of rdd triple mutants, which lack RDD, indicating that Cd-induced DNA methylation is associated with the inhibition of RDD. Interestingly, the elevation in DNA methylation in rdd conferred a higher tolerance against Cd stress and improved cellular Fe nutrition in the root tissues. In addition, lowering the Fe supply abolished improved Cd tolerance due to the lack of RDD in rdd. Together, these data suggest that the inhibition of RDD-mediated DNA demethylation in the roots by Cd would in turn enhance plant tolerance to Cd stress by improving Fe nutrition through a feedback mechanism.

Automated detection and quantification of COVID-19 pneumonia: CT imaging analysis by a deep learning-based software.

BACKGROUND: The novel coronavirus disease 2019 (COVID-19) is an emerging worldwide threat to public health. While chest computed tomography (CT) plays an indispensable role in its diagnosis, the quantification and localization of lesions cannot be accurately assessed manually. We employed deep learning-based software to aid in detection, localization and quantification of COVID-19 pneumonia. METHODS: A total of 2460 RT-PCR tested SARS-CoV-2-positive patients (1250 men and 1210 women; mean age, 57.7 ± 14.0 years (age range, 11-93 years) were retrospectively identified from Huoshenshan Hospital in Wuhan from February 11 to March 16, 2020. Basic clinical characteristics were reviewed. The uAI Intelligent Assistant Analysis System was used to assess the CT scans. RESULTS: CT scans of 2215 patients (90%) showed multiple lesions of which 36 (1%) and 50 patients (2%) had left and right lung infections, respectively (> 50% of each affected lung's volume), while 27 (1%) had total lung infection (> 50% of the total volume of both lungs). Overall, 298 (12%), 778 (32%) and 1300 (53%) patients exhibited pure ground glass opacities (GGOs), GGOs with sub-solid lesions and GGOs with both sub-solid and solid lesions, respectively. Moreover, 2305 (94%) and 71 (3%) patients presented primarily with GGOs and sub-solid lesions, respectively. Elderly patients (≥ 60 years) were more likely to exhibit sub-solid lesions. The generalized linear mixed model showed that the dorsal segment of the right lower lobe was the favoured site of COVID-19 pneumonia. CONCLUSION: Chest CT combined with analysis by the uAI Intelligent Assistant Analysis System can accurately evaluate pneumonia in COVID-19 patients.

[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.

[The role of cellular retinol-binding protein-1 on pulmonary structural remodeling after rat myocardial infarction].

OBJECTIVES: To observe the expression of cellular retinol-binding protein-1 (CRBP-1) in pulmonary tissues after rat myocardial infarction (MI) and uncover the role of CRBP-1 on the pulmonary structural remodeling. METHODS: MI was produced in male Wistar rats by left coronary ligation. Rats were sacrificed to obtain the lung at the 3(rd), 6(th), 15(th), 30(th), and 45(th) day after operation. After weighted, the rat lungs were fixed in 4% formalin and embedded in paraffin. Sections were cut and stained with hematoxylin and eosin (HE), Masson's trichrome (MT), rabbit anti-CRBP-1 antibody. RESULTS: Thirty-nine Wistar rats survived and developed MI. Pulmonary tissue sections with HE and MT staining showed a remarkable lung structural remodeling. The content of pulmonary tissue collagen at the 30(rd) was higher than in the sham group (8.4% ± 3.6 vs 4.5% ± 2.6, P < 0.001). CRBP-1 expression was detected on the alveolar septa at the 3(rd) day after operation, and peaked at the 15(th) day (43.8 ± 7.4). Then the CRBP-1 expression decreased, and arrived at the level of the 3(rd) day. CONCLUSIONS: We demonstrate that CRBP-1 is expressed temporal on the alveolar septa after rat MI. It indicates a potential relationship between CRBP-1 and lung structure remodeling process after MI.

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.

Phloem iron remodels root development in response to ammonium as the major nitrogen source.

Plants use nitrate and ammonium as major nitrogen (N) sources, each affecting root development through different mechanisms. However, the exact signaling pathways involved in root development are poorly understood. Here, we show that, in Arabidopsis thaliana, either disruption of the cell wall-localized ferroxidase LPR2 or a decrease in iron supplementation efficiently alleviates the growth inhibition of primary roots in response to NH4+ as the N source. Further study revealed that, compared with nitrate, ammonium led to excess iron accumulation in the apoplast of phloem in an LPR2-dependent manner. Such an aberrant iron accumulation subsequently causes massive callose deposition in the phloem from a resulting burst of reactive oxygen species, which impairs the function of the phloem. Therefore, ammonium attenuates primary root development by insufficiently allocating sucrose to the growth zone. Our results link phloem iron to root morphology in response to environmental cues.

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.

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.

Roles of ezrin in the growth and invasiveness of esophageal squamous carcinoma cells.

Ezrin, which crosslinks the cytoskeleton and plasma membrane, is involved in the growth and metastatic potential of cancer cells. Ezrin expression in esophageal squamous cell carcinoma (ESCC) was described recently, but its roles and the underlying mechanism(s) remain unclear. In our study, we first showed that ezrin in ESCC cell is expressed in the nucleus as well as in the cytoplasm and plasma membrane. Then, by using RNAi, we revealed that interference of ezrin expression suppressed the growth, adhesion and invasiveness of ESCC cells. Tumorigenesis experiments revealed that ezrin may directly regulate tumor formation in vivo. To explore the molecular mechanisms through which ezrin contributes to the proliferation and invasiveness of ESCC cells, we used cDNA microarrays to analyze ezrin knockdown cells and the control cells; of 39,000 genes examined, 297 were differentially expressed upon ezrin knockdown, including some proliferation- and invasiveness-related genes such as ATF3, CTGF and CYR61. Furthermore, pathway analysis showed that ezrin knockdown led to decreased activation of the TGF-beta and MAPK pathways, and ezrin-mediated cell invasiveness alteration was dependent on the activation of these pathways. Finally, immunohistochemical staining on 80 ESCC specimens and 50 normal esophageal mucosae revealed that the expression levels of 3 altered genes involved in the regulation of cell proliferation and tumor metastasis, including CTGF, CYR61 and ATF3, were altered in ESCCs, and their expression pattern correlated with ezrin expression. Taken together, we propose that ezrin might function in the growth and invasiveness of ESCC cells through the MAPK and TGF-beta pathways.

[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.

[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.

[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.

[Effect of Eletroacupuncture Intervention on Insulin Resistance, Lipid Metabolic Disorder and Expression of Hepatic SREBP-1 c and Fatty Acid Synthase Proteins in Rats with Hyperlipidemia].

OBJECTIVE: To observe the effect of electroacupuncture(EA) of "Fenglong" (ST 40) and "Sanyinjiao" (SP 6) on lipid metabolic disorder, insulin resistance (IR) and expression of sterol regulatory element blinding protein-1 (SREBP-1) c and fatty acid synthase (FAS) proteins in the liver tissue in hyperlipidemia rats with 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 group). The IR model was established by feeding the rat with high-fat diet. Rats of the medication group were treated by intragastric administration of pioglitazone (10 mL/kg). For rats of the EA group, EA (2 Hz/100 Hz，1 mA) was applied to bilateral ST 40 and SP 6, once daily for 14 days. The insulin sensitivity index (ISI) was assessed by calculating 60－120 min glucose infusion rate (GIR 60－120) with euglycemic hyperinsulinemic clamp in reference to Kraegen's and colleagues' methods. Fasting blood samples (10 mL) were collected and analyzed for fasting blood glucose (FBG) using enzyme method, serum fasting insulin(FINS) using ELISA, free fatty acid(FFA) using spectrophotometry, and total triglyceride(TG) and total cholesterol(TC) employing glycerine phosphate oxidase peroxidase (GPO-PAP) assay, low density lipoprotein(LDL), high density lipoprotein(HDL) levels using combined filiter paper activity and lipase activity methods, respectively. The IR level was assessed by calculating homeostatic model assessment of insulin resistance (HOMA-IR) using the formula (FBG×FINS)/22.5. The expression levels of SREBP-1 c and FAS proteins in the liver tissue were detected by Western blot. RESULTS: Following modeling, the GIR 60－120 and serum HDL were significantly decreased(P<0.01), and the HOMA-IR, serum FBG, FINS, FFA, TG, TC and LDL, and the expression levels of hepatic SREBP-1 c and FAS proteins were significantly increased in comparison with the blank control group(P<0.01). After the intervention, the decreased GIR 60－120 and serum HDL levels were considerably up-regulated (P<0.01), and the increased FBG, FINS, FFA, TG, TC and LDL, and hepatic SREBP-1 c and FAS protein levels were notably down-regulated in both EA and medication groups relevant to the model group (P<0.05, P<0.01). No significant differences were found between the EA and medication groups in all the indexes mentioned above (P>0.05). CONCLUSION: EA intervention is able to improve the disorder of lipid metabolism of IR rats, which may be associated with its effects in reducing the expression of SREBP-1 c and FAS proteins and in lowering the synthesis of fatty acid.

Sulfide alleviates cadmium toxicity in Arabidopsis plants by altering the chemical form and the subcellular distribution of cadmium.

Several sulfur compounds are thought to play important roles in the plant tolerance to cadmium (Cd), but the role of inorganic sulfide in Cd tolerance remains largely unknown. In this study, we found that Cd exposure increased the accumulation of soluble sulfide in Arabidopsis plants. When exogenous sulfide, in the form of NaHS, was foliarly applied, Cd-induced growth inhibition and oxidative stress were alleviated. In addition, although the foliar application of sulfide did not affect the total Cd levels, it significantly decreased the soluble Cd fractions in plants. Furthermore, foliar applications of sulfide decreased Cd distribution in the cytoplasm and organelles, but increased Cd retention in the cell wall, which is a less sensitive compartment. These results suggest that the Cd-induced accumulation of soluble sulfide alleviates Cd toxicity in plants by inactivating Cd and sequestering it into the cell wall.

[Acupoints Selection Exploring for Acupuncture Treatment of Insulin Resistance Based on Data Mining].

OBJECTIVE: To analyze the characteristics and rules of acupoint selection for acupuncture treatment of insulin resistance. METHODS: Data collections were conducted by searching references on acupuncture treatment of insulin resistance in PubMed, CNKI, VIP data base from 1991 to 2016, and acupuncture prescription data base for acupuncture treatment of insulin resistance was established. Data mining was applied to analyze the characteristics and rules of acupoint selection. RESULTS: A total of 64 papers were recruited, and 73 acupoints were selected in these papers. It was found that the acupoints as Zusanli (ST 36), Sanyinjiao (SP 6), Fenglong (ST 40) and Taichong (LR 3) were used with highest frequencies. All acupoints selected distributed in 13 meridians, especially Foot Yangming Stomach Meridian, Foot Taiyin Bladder Meridian, and Conception Vessel with a total frequency of 58.07%. The special acupoints including five-shu points, eight confluent points and back-shu points, accounted for 56.71%. CONCLUSIONS: This study excavated the regular acupoint selection and acupoints compatibility for acupuncture treatment in patients with insulin resistance, giving evidence based confirming and direction for acupuncture clinical treatment.

[Episodic central nervous system symptoms with reversible white matter involvement in Chinese patients with X-linked Charcot-Marie-Tooth disease and literatures review].

OBJECTIVE: To analyze the phenotype and genotype of CMTX1 patients with episodic transient reversible white matter involvement, and delineate the features of brain MRI in the episode and the possible mechanisms. METHOD: Three Chinese probands and their family members were sequenced in the coding regions of GJB1. With the other 16 reported CMTX1 patients with episodic transient reversible white matter involvement, the clinical feature of the episodic central nervous system symptoms and the genotypes were reviewed. RESULT: Missense mutations in GJB1 were identified in all 3 probands. In 19 patients with transient reversible white matter involvement, the episodes were manifested as weakness of the limbs, dysarthria, and dysphagia, without disturbance of consciousness or seizures. The episodes lasted for 13 hours (10 min-72 hours) with complete remission in all patients; There were multiple episodes in 9 patients. During the episode, brain MRI showed symmetrical high signals in T2 weighted, Flair and DWI images in periventricular white matter, with predominance in posterior region including splenium of corpus callosum. These changes in imaging were most prominent during or within 1 week after the clinical episode.Significant improvements occurred within 1 month, with complete remission within 4-6 months.No specific locations of mutant amino acids in GJB1 protein were found in these patients with episodic transient reversible white matter involvement. CONCLUSION: Episodic transient reversible white matter involvement may present in a small number of patients with CMTX1. Transient edema of oligodendrocytes due to the dysfunction of gap junction may be involved in the pathogenesis. There is no correlation between the location of the mutant amino acids in GJB1 and the occurrence of the episodes.

The histone deacetylase inhibitor trichostatin A induces cell cycle arrest and apoptosis in colorectal cancer cells via p53-dependent and -independent pathways.

Many chemotherapeutic agents induce apoptosis via a p53-dependent pathway. However, up to 50% of human cancers have p53 mutation and loss of p53 function. Histone deacetylase inhibitors (HDACIs) are emerging as a potentially important new class of anticancer agents. Here, we report that, Trichostatin A (TSA), a pan-HDAC inhibitor, could induce G2/M cell cycle arrest and apoptosis in both colorectal cancer cell lines with wild-type p53 (HT116 cells) and mutant p53 (HT29 cells), although HCT116 cells had more apoptotic cells than HT29 cells. TSA induces apoptosis in both cell lines via the mitochondrial pathway as indicated by decrease of the mitochondrial membrane potential (MMP) and activation of caspase-3. Additionally, TSA induces expression of the pro-apoptotic protein Bax and decreases the expression of the anti-apoptotic proteins Bcl-2 and Bcl-xL in both cell lines. Bax knockdown by siRNA significantly impaired TSA-induced apoptosis in both cell lines. These data suggest that TSA induces G2/M cell cycle arrest and Bax-dependent apoptosis in colorectal cancer cells (HCT116 cells and HT29 cells) by both p53-dependent and -independent mechanisms. However, cells with normal p53 function are more sensitive to TSA-induced apoptosis.

[The method and influencing factors of alkalescency protein purification from bitter melon (Momordica charantia) seeds by polyacrylicacid precipitation].

In this study, polyacrylicacid precipitation alkalescence protein from Momordica charantia L. seeds was studied, and the effect of conditions on experiment was also evaluated. Isoelectric precipitation is achieved by adjusting the pH of a protein solution and is based on that a protein's solubility is at minimum at its pI. The sample was titrated to pH 6.0 with citric acid, and 14.62% proteins were precipitated. With hydrochloric acid to pH 4.0, 32.49% proteins were precipitated. With the acetic acid to pH 6.0 and pH 4.0, 26.17% and 38.72% proteins were precipitated, respectively. In the 1 mL Bitter melon seeds extraction(pH 4.0) adjusted by acetic acid, hydrochloric acid and citric acid, the optimum dosage of PAA (1%) precipiting alkalescency protein (pl 8.65-9.30) was 100 microL, 120 microL and 100 microL, respectively. The respective extraction (1mL) was titrated to pH 5.0, pH 4.0, and pH 3.0 by acetic acid. After isoelectric precipitation, the PAA precipitation protein was performed. When concentration of PAA (1%) was 160 microL/mL, the protein decreased in the supernatant was 33.77% at pH 5.0, and 43.56% at pH 3.0. When concentration of PAA (1%) was 120 microL/mL, the protein decreased in the supernatant was 30.83% at pH 4.0. PAA-Protein complex could redissolve in alkaline conditions (pH > 9.0) and the protein most easilly redissolved when the NaCL was 3.0%. The bitter melon seeds extraction after PAA purification flowed through the Sephadex G-75 columns. The peaks I and II were obtained after 175 min and 300 min, respectively. SDS-PAGE and IEF analysis showed that the molecule weight from peaks I was 30 kD with pI 9.5, peaks II 10 kD with pI 9.3.