Reducing myocardial infarction by combination of irisin and Dendrobium nobile Lindl through inhibiting nod-like receptor protein-3-related pyroptosis and activating PINK1/Parkin-mitophagy during aging.

Aging, a crucial risk factor for ischemic heart disease, has negative impacts on cardioprotective mechanisms. As such, there is still an unmet requirement to explore potential therapies for improving the outcomes of myocardial ischemia/reperfusion (IR) injury in elderly subjects. Here, we aimed to confirm the cardioprotective function of irisin/Dendrobium nobile Lindl (DNL) combination therapy against myocardial IR injury in aged rats, with a focus on the involvement of pyroptosis and mitophagy. Male aged Wistar rats (22-24 months old, 400-450 g; n = 54) underwent myocardial IR or sham surgery. Before IR operation, rats were pretreated with irisin (0.5 mg/kg, intraperitoneally) and/or DNL (80 mg/kg, orally) for 1 or 4 weeks, respectively, at corresponding groups. Cardiac function, lactate dehydrogenase (LDH) and cardiac-specific isoform of troponin-I (cTn-I) levels, the expression of proteins involved in pyroptosis (nod-like receptor protein-3 (NLRP3), apoptosis-associated speck-like protein, c-caspase-1, and GSDMD-N) and mitophagy (PINK1 and Parkin), and pro-inflammatory cytokines levels were evaluated after 24 h of reperfusion. Irisin/DNL combined therapy significantly restored cardiac function and decreased LDH and cTn-I levels. It also downregulated pyroptosis-related proteins, upregulated PINK1 and Parkin, and decreased pro-inflammatory cytokines secretion. Pretreatment with Mdivi-1, as mitophagy inhibitor, abolished the cardioprotective action of dual therapy. This study revealed the cardioprotective effects of irisin/DNL combination therapy against IR-induced myocardial injury in aged rats, and also showed that the mechanism might be associated with suppression of NLRP3-related pyroptosis through enhancing the activity of the PINK1/Parkin mitophagy. This combination therapy is worthy of further detailed studies due to its potential to alleviate myocardial IR injury upon aging.

Durability Study of Thermal Transfer Printed Textile Electrodes for Wearable Electronic Applications.

Textile-based electronics hold great promise because they can endow wearable devices with soft and comfortable characteristics. However, the inherent porosity and fluffiness of fabrics result in high surface roughness, which presents great challenges in the manufacture of high-performance fabric electrodes. In this work, we propose a thermal transfer printing method to address the above challenges, in which electrodes or circuits of silver flake/thermoplastic polyurethane (TPU) composites are prefabricated on a release film by coating and laser engraving and then laminated by hot-pressing to a variety of fabrics and textiles. This universal and scalable production technique enables fabric electrodes to be made without compromising the original wearability, washability, and stretchability of textiles. The prepared fabric electrodes exhibit high conductivity (5.48 × 104 S/cm), high adhesion (≥1750 N/m), good abrasion/washing resistance, high patterning resolution (∼40 µm), and good electromechanical performance up to 50% strain. To demonstrate the potential applications, we developed textile-based radio frequency identification (RFID) tags for remote identification and a large-sized heater for wearable thermotherapy. More importantly, the solvent-free thermal transfer printing technology developed in this paper enables people to DIY interesting flexible electronics on clothes with daily tools, which can promote the commercial application of smart textile-based electronics.

Comparison of femoral nerve block and acupuncture analgesia for acute preoperative pain in elderly patients with femoral neck fracture: a retrospective study.

OBJECTIVE: This study was to compare the efficacy of femoral nerve block (FNB) and acupuncture for acute preoperative pain in patients with femoral neck fracture (FNF). METHODS: From June 2017 to June 2019, 130 patients with FNF were included in this study. Sixty-six patients received FNB treatment (FNB group) and sixty-four patients received acupuncture treatment (Acupuncture group). The clinical information, visual analog scale (VAS) scores, nursing quality scores, sleep quality scores, delirium numbers, and perioperative complications were collected and compared between the 2 groups. RESULTS: The resting VAS score and the exercise VAS score decreased after FNB or acupuncture in both groups. Thirty minutes after analgesia, the resting VAS scores in the FNB group and the acupuncture group were 27.3±8.0 and 27.9±7.8, respectively (P=0.67); while exercise VAS scores were 60.2±10.4 and 59.5±9.8, respectively (P=0.73). In addition, there was no statistical difference in the VAS score between the two groups on day 1 and day 2 after admission. There was no statistical difference in nursing quality, sleep rhythm disorder, sleep quality, or times of mental disorder between the two groups. CONCLUSION: FNB analgesia and acupuncture analgesia are safe and effective for the control of acute preoperative pain in senile patients with femoral neck fracture. Both methods have good analgesic effects, which can improve nursing and sleep quality, and reduce the incidence of delirium. As a traditional Chinese medicine method, acupuncture analgesia can effectively manage the acute preoperative pain in senile femoral neck fracture patients.

CO2 Laser Moxibustion for Knee Osteoarthritis: Study Protocol for A Multicenter, Double-blind, Randomized Controlled Trial.

BACKGROUND: Knee osteoarthritis (OA) is a major cause of disability among the older adults. Few treatments are safe and effective. Moxibustion is commonly used in treating knee OA in Chinese medicine (CM). CO2 Laser moxibustion device is a substitute for traditional moxibustion, which mimics the effects of traditional moxibustion. More data are needed to support its application in knee OA. OBJECTIVE: ObjectiveThe trial aims to assess the effect and safety of CO2 laser moxibustion in patients with knee osteoarthritis compared with a sham control. METHODS: This is a protocol for a multicenter, randomized, double-blind, placebo-controlled trial. A total of 392 participants were recruited and assigned to the CO2 laser moxibustion group and sham laser moxibustion group with a 1:1 ratio at 6 outpatient clinics in Shanghai, China. Participants in both groups received treatment at the affected knee(s) at the acupuncture point Dubi (ST 35) and an Ashi point. There were 3 sessions per week for 4 weeks, and an additional 20-week follow-up. Primary outcomes were changes in the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain scores at week 4. Secondary outcomes were WOMAC function score, stiffness score and overall score, VAS pain, Short-Form heath survey (SF-36), and patients' global assessment. The serum levels of cytokines involved in progress of knee OA were explored. Safety was assessed during the whole trial. Masking effectiveness was assessed by both participants and treatment providers.This is a protocol for a multicenter, randomized, double-blind, placebo-controlled trial. A total of 392 participants were recruited and assigned to the CO2 laser moxibustion group and sham laser moxibustion group with a 1:1 ratio at 6 outpatient clinics in Shanghai, China. Participants in both groups received treatment at the affected knee(s) at the acupuncture point Dubi (ST 35) and an Ashi point. There were 3 sessions per week for 4 weeks, and an additional 20-week follow-up. Primary outcomes were changes in the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain scores at week 4. Secondary outcomes were WOMAC function score, stiffness score and overall score, VAS pain, Short-Form heath survey (SF-36), and patients' global assessment. The serum levels of cytokines involved in progress of knee OA were explored. Safety was assessed during the whole trial. Masking effectiveness was assessed by both participants and treatment providers. DISCUSSION: CO2 laser moxibustion device, designed as a substitute for CM moxibustion, is easy to use and control with no choking smoke and smell, and is a plausible method for double-blind research. This study would provide rigorous evidence for the effect and safety of CO2 laser moxibustion in treating knee OA (Trial registration No.: ISRCTN15030019).

miR-541 potentiates the response of human hepatocellular carcinoma to sorafenib treatment by inhibiting autophagy.

OBJECTIVE: Autophagy participates in the progression of hepatocellular carcinoma (HCC) and the resistance of HCC cells to sorafenib. We investigated the feasibility of sensitising HCC cells to sorafenib by modulating miR-541-initiated microRNA-autophagy axis. DESIGN: Gain- and loss-of-function assays were performed to evaluate the effects of miR-541 on the malignant properties and autophagy of human HCC cells. Autophagy was quantified by western blotting of LC3, transmission electron microscopy analyses and confocal microscopy scanning of mRFP-GFP-LC3 reporter construct. Luciferase reporter assays were conducted to confirm the targets of miR-541. HCC xenograft tumours were established to analyse the role of miR-541 in sorafenib-induced lethality. RESULTS: The expression of miR-541 was downregulated in human HCC tissues and was associated with malignant clinicopathologic phenotypes, recurrence and survival of patients with HCC. miR-541 inhibited the growth, metastasis and autophagy of HCC cells both in vitro and in vivo. Prediction software and luciferase reporter assays identified autophagy-related gene 2A (ATG2A) and Ras-related protein Rab-1B (RAB1B) as the direct targets of miR-541. Consistent with the effects of the miR-541 mimic, inhibition of ATG2A or RAB1B suppressed the malignant phenotypes and autophagy of HCC cells. Furthermore, siATG2A and siRAB1B partially reversed the enhancement of the malignant properties and autophagy in HCC cells mediated by the miR-541 inhibitor. More interestingly, higher miR-541 expression predicted a better response to sorafenib treatment, and the combination of miR-541 and sorafenib further suppressed the growth of HCC cells in vivo compared with the single treatment. CONCLUSIONS: Dysregulation of miR-541-ATG2A/RAB1B axis plays a critical role in patients' responses to sorafenib treatment. Manipulation of this axis might benefit survival of patients with HCC, especially in the context of the highly pursued strategies to eliminate drug resistance.

Allele-selective lowering of mutant HTT protein by HTT-LC3 linker compounds.

Accumulation of mutant proteins is a major cause of many diseases (collectively called proteopathies), and lowering the level of these proteins can be useful for treatment of these diseases. We hypothesized that compounds that interact with both the autophagosome protein microtubule-associated protein 1A/1B light chain 3 (LC3)1 and the disease-causing protein may target the latter for autophagic clearance. Mutant huntingtin protein (mHTT) contains an expanded polyglutamine (polyQ) tract and causes Huntington's disease, an incurable neurodegenerative disorder2. Here, using small-molecule-microarray-based screening, we identified four compounds that interact with both LC3 and mHTT, but not with the wild-type HTT protein. Some of these compounds targeted mHTT to autophagosomes, reduced mHTT levels in an allele-selective manner, and rescued disease-relevant phenotypes in cells and in vivo in fly and mouse models of Huntington's disease. We further show that these compounds interact with the expanded polyQ stretch and could lower the level of mutant ataxin-3 (ATXN3), another disease-causing protein with an expanded polyQ tract3. This study presents candidate compounds for lowering mHTT and potentially other disease-causing proteins with polyQ expansions, demonstrating the concept of lowering levels of disease-causing proteins using autophagosome-tethering compounds.

Proteomic Investigations of Transcription Factors Critical in Geniposide-Mediated Suppression of Alcoholic Steatosis and in Overdose-Induced Hepatotoxicity on Liver in Rats.

Alcoholic steatosis is one of the most prevalent forms of liver disease, and appropriate insight and application of anti-steatosis drugs must be considered. Geniposide, the major active constituent of the Gardenia jasminoides (Ellis) fruit, has been commonly used as a traditional herbal medicine for the treatment of liver diseases. However, its hepatoprotective effect on alcoholic steatosis has not been reported. Moreover, geniposide overdose-induced hepatotoxicity was demonstrated. Hence, its therapeutic effects and overdose-induced hepatotoxicity in rat models along with corresponding targets, especially the targets of transcription factors (TFs), were systematically investigated in this study by using a concatenated tandem array of consensus TF response elements. The results indicate that geniposide can attenuate alcoholic steatosis and liver injury by enhancing the transcriptional activities of peroxisome proliferator-activated receptor-α and hepatocyte nuclear factors 1α and 4α, while geniposide overdose perturbs other TFs. In addition, therapeutic doses and overdoses of geniposide have differentiated target TFs. This study is the first to provide a systematic insight into the difference of critical transcription factors between the actions of therapeutic doses and overdoses of geniposide, as well as much-needed attention to the important topic of alcoholic liver disease therapy.

[Effect of Dry Needling Stimulation of Myofascial Trigger Point on Sample Entropy of Electromyography of Gastrocnemius Injured Site in Rats].

OBJECTIVE: To attempt to establish an objective quantitative indicator to characterize the trigger point activity, so as to evaluate the effect of dry needling on myofascial trigger point activity. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into blank control group, dry needling (needling) group, stretching exercise (stretching) group and needling plus stretching group (n＝6 per group). The chronic myofascial pain (trigger point) model was established by freedom vertical fall of a wooden striking device onto the mid-point of gastrocnemius belly of the left hind-limb to induce contusion, followed by forcing the rat to make a continuous downgrade running exercise at a speed of 16 m/min for 90 min on the next day which was conducted once a week for 8 weeks. Electromyography (EMG) of the regional myofascial injured point was monitored and recorded using an EMG recorder via electrodes. It was considered success of the model if spontaneous electrical activities appeared in the injured site. After a 4 weeks' recovery, rats of the needling group were treated by filiform needle stimulation (lifting-thrusting-rotating) of the central part of the injured gastrocnemius belly (about 10 mm deep) for 6 min, and those of the stretching group treated by holding the rat's limb to make the hip and knee joints to an angle of about 180°, and the ankle-joint about 90° for 1 min every time, 3 times altogether (with an interval of 1 min between every 2 times). The activity of the trigger point was estimated by the sample entropy of the EMG signal sequence in reference to Richman's and Moorman's methods to estimate the curative effect of both needling and exercise. RESULTS: After the modeling cycle, the mean sample entropies of EMG signals was significantly decreased in the model groups (needling group [0.034±0.010], stretching group [0.045±0.023], needling plus stretching group [0.047±0.034]) relevant to the blank control group (0.985±0.196, P<0.01). After the treatment, the mean sample entropy of EMG signals was evidently increased in both needling (0.819±0.088), stretching (0.532±0.25) and needling plus stretching (0.810±0.117) groups (P<0.01). The mean sample entropy of the needling and needling plus stretching groups were significantly higher than that of the stretching group (P<0.01), without remarkable difference between the two needling groups in the mean sample entropy (P>0.05), suggesting a better efficacy of dry needling in easing trigger point activity. CONCLUSION: Dry needling is able to relieve myofascial trigger point activity in rats, which is better than that of simple passive stretching therapy.

A novel ventricular restraint device (ASD) repetitively deliver Salvia miltiorrhiza to epicardium have good curative effects in heart failure management.

A novel ventricular restraint is the non-transplant surgical option for the management of an end-stage dilated heart failure (HF). To expand the therapeutic techniques we design a novel ventricular restraint device (ASD) which has the ability to deliver a therapeutic drug directly to the heart. We deliver a Traditional Chinese Medicine (TCM) Salvia miltiorrhiza (Danshen Zhusheye) through active hydraulic ventricular support drug delivery system (ASD) and we hypothesize that it will show better results in HF management than the restraint device and drug alone. SD rats were selected and divided into five groups (n=6), Normal, HF, HF+SM (IV), HF+ASD, HF+ASD+SM groups respectively. Post myocardial infarction (MI), electrocardiography (ECG) showed abnormal heart function in all groups and HF+ASD+SM group showed a significant therapeutic improvement with respect to other treatment HF, HF+ASD, and HF+SM (IV) groups on day 30. The mechanical functions of the heart such as heart rate, LVEDP, and LVSP were brought to normal when treated with ASD+SM and show significant (P value<0.01) compared to other groups. BNP significantly declines in HF+ASD+SM group animals compared with other treatment groups. Masson's Trichrome staining was used to study histopathology of cardiac myocytes and quantification of fibrosis was assessed. The large blue fibrotic area was observed in HF, HF+ASD, and HF+SM (IV) groups while HF+ASD+SM showed negligible fibrotic myocyte at the end of study period (30days). This study proves that novel ASD device augments the therapeutic effect of the drug and delivers Salvia miltiorrhiza to the cardiomyocytes significantly as well as provides additional support to the dilated ventricle by the heart failure.

An integrated approach to identify critical transcription factors in the protection against hydrogen peroxide-induced oxidative stress by Danhong injection.

Oxidative stress plays a vital role in many pathological processes of the cardiovascular diseases. However, the underlying mechanism remains unclear, especially on a transcription factor (TF) level. In this study, a new method, concatenated tandem array of consensus transcription factor response elements (catTFREs), and an Illumina-based RNA-seq technology were integrated to systematically investigate the role of TFs in hydrogen peroxide (H2O2)-induced oxidative stress in cardiomyocytes; the damage was then rescued by Danhong injection (DHI), a Chinese standardized product approved for cardiovascular diseases treatment. The overall gene expression revealed cell apoptosis and DNA repair were vital for cardiomyocytes in resisting oxidative stress. By comprehensively integrating the transcription activity of TFs and their downstream target genes, an important TFs-target network were constructed and 13 TFs were identified as critical TFs in DHI-mediated protection in H2O2-induced oxidative stress. By using the integrated approach, seven TFs of these 13 TFs were also identified in melatonin-mediated protection in H2O2-induced damage. Furthermore, the transcription activity of DNA-(apurinic or apyrimidinic site) lyase (Apex1), Myocyte-specific enhancer factor 2D (Mef2d) and Pre B-cell leukemia transcription factor 3 (Pbx3) was further verified in pluripotent stem cell-derived cardiomyocytes. This research offers a new understanding of cardiomyocytes in response to H2O2-induced oxidative stress and reveals additional potential therapeutic targets. The combination of two parallel omics datasets (corresponding to the transcriptome and proteome) can reduce the noise in high-throughput data and reveal the fundamental changes of the biological process, making it suitable and reliable for investigation of critical targets in many other complicated pathological processes.

Chinese medicine for outcomes in colorectal cancer patients: A retrospective clinical study.

OBJECTIVE: To investigate the effect of Chinese medicine (CM) on survival of patients with stage II and III colorectal cancer (CRC). METHODS: A total of 295 patients who received chemotherapy were assigned to Group 1. The other 171 patients received the same chemotherapy treatment combined with the usage of CM Jianpi Jiedu Formula (, JPJD) for more than 3 months (Group 2). Patients' survival time, relapse and metastasis, and cause of death were observed. Cox proportional hazard regression models were established for the analysis of the effect of independent factors on the survival prognosis of patients with CRC. RESULTS: The survival rate of patients in Group 2 was higher than that of Group 1 (P<0.05). Compared with Group 1, the mean survival time was prolonged by 5.594 months and the median survival time was prolonged by 6 months in Group 2 (P=0.004). Cox regression analysis indicated that CM combined with chemotherapy provided signifificant protective effect, as observed with the improvements in the survival rates of CRC patients (P<0.01). CONCLUSION: CM can improve the survival rate in patients with stage II and III CRC.

Systematic investigation of transcription factors critical in the protection against cerebral ischemia by Danhong injection.

Systematic investigations of complex pathological cascades during ischemic brain injury help to elucidate novel therapeutic targets against cerebral ischemia. Although some transcription factors (TFs) involved in cerebral ischemia, systematic surveys of their changes during ischemic brain injury have not been reported. Moreover, some multi-target agents effectively protected against ischemic stroke, but their mechanisms, especially the targets of TFs, are still unclear. Therefore, a comprehensive approach by integrating network pharmacology strategy and a new concatenated tandem array of consensus transcription factor response elements method to systematically investigate the target TFs critical in the protection against cerebral ischemia by a medication was first reported, and then applied to a multi-target drug, Danhong injection (DHI). High-throughput nature and depth of coverage, as well as high quantitative accuracy of the developed approach, make it more suitable for analyzing such multi-target agents. Results indicated that pre-B-cell leukemia transcription factor 1 and cyclic AMP-dependent transcription factor 1, along with six other TFs, are putative target TFs for DHI-mediated protection against cerebral ischemia. This study provides, for the first time, a systematic investigation of the target TFs critical to DHI-mediated protection against cerebral ischemia, as well as reveals more potential therapeutic targets for ischemic stroke.

The moderating and mediating roles of self-acceptance and tolerance to others in the relationship between mindfulness and subjective well-being.

This study explored the moderating and mediating influences of self-acceptance and tolerance to others in the relationship between mindfulness and subjective well-being. In total, 301 (130 males) university students completed the Five-Facet Mindfulness Questionnaire, Index of Well-being, Self-acceptance Questionnaire, and Tolerance Scale. The results showed that the positive link between mindfulness and subjective well-being was significantly mediated by self-acceptance only. Tolerance played a moderating role. The implications of the results for relevant research and mindfulness training were discussed.

Gingerol activates noxious cold ion channel TRPA1 in gastrointestinal tract / 中国天然药物

TRPA1 channels are non-selective cation channels that could be activated by plant-derived pungent products, including gingerol, a main active constituent of ginger. Ginger could improve the digestive function; however whether ginger improves the digestive function through activating TRPA1 receptor in gastrointestinal tract has not been investigated. In the present study, gingerol was used to stimulate cell lines (RIN14B or STC-1) while depletion of extracellular calcium. TRPA1 inhibitor (rethenium red) and TRPA1 gene silencing via TRPA1-specific siRNA were also used for mechanistic studies. The intracellular calcium and secretion of serotonin or cholecystokinin were measured by fura-2/AM and ELISA. Stimulation of those cells with gingerol increased intracellular calcium levels and the serotonin or cholecystokinin secretion. The gingerol-induced intracellular calcium increase and secretion (serotonin or cholecystokinin) release were completely blocked by ruthenium red, EGTA, and TRPA1-specific siRNA. In summary, our results suggested that gingerol derived from ginger might improve the digestive function through secretion releasing from endocrine cells of the gut by inducing TRPA1-mediated calcium influx.

[Alisma versus Gliclazide in the Treatment of Primary Diabetes in Goto-Kakizaki Rats].

OBJECTIVE: To compare the roles of alisma and gliclazide in the treatment of diabetes in Goto-Kakizaki (GK) rats. METHODS: GK rats were randomly divided into alisma group, gliclazide group, and blank group, and Wistar rats were used as the normal group. After two weeks of treatment, body weight, food intake,fasting glucose, impaired glucose tolerance, and other indicators were measured. RESULTS: The body weight increased after the treatment in the normal group,blank group,and gliclazide group [(241.3 ± 7.0)g vs.(263.5 ± 11.1)g, (242.8 ± 7.1)g vs.(267.9 ± 16.8)g, (243.9 ± 12.2)g vs.(277.9 ± 9.8)g, P<0.05] but decreased in alisma group [(244.6 ± 9.2)g vs.(227.9 ± 13.7)g, P<0.05]. The food intake showed no significant change before and after administration among different groups(P>0.05). Fasting glucose was significantly lower in normal group than in control group,alisma group,and gliclazide group [(4.8 ± 0.2) mmol/L vs.(8.2 ± 1.4) mmol/L,(8.1 ± 0.6) mmol/L, (8.1 ± 0.9)mmol/L, P<0.05] one week after drug administration; it was not significantly different among blank group,alisma group,and gliclazide group before drug administration (P>0.05); however, it significantly decreased in alisma group and gliclazide group two weeks after administration [(6.9 ± 0.7) mmol/L vs.(8.1 ± 0.6) mmol/L; (5.8 ± 0.5) mmol/L vs.(8.1 ± 0.9) mmol/L, P<0.05]; compared with the blank group, the fasting glucose was significantly lower in the alisma group and gliclazide group,and it was also significantly different between these two groups [(6.9 ± 0.7) mmol/L vs.(8.8 ± 0.6) mmol/L,(5.8 ± 0.5)mmol/L vs.(8.8 ± 0.6)mmol/L, (6.9 ± 0.7) mmol/L vs.(5.8 ± 0.5)mmol/L, P<0.05]. Compared with the normal group,glucose tolerance was abnormal in blank group,alisma group,and gliclazide group;after two weeks of treatment,glucose tolerance was significantly improved in alisma group (P<0.05); compared with the pretreatment level and that in the blank group,the glucose tolerance in gliclazide group showed no significant difference (P> 0.05). CONCLUSIONS: Both alisma and gliclazide monotherapy is effective in lowering fasting blood glucose. As a single-target drug,gliclazide has stronger effecacy in lowering fasting glucose. However, alisma, as a mixture, can also control weight and improve glucose intolerance.

Enrichment and Characterization of a Psychrotolerant Consortium Degrading Crude Oil Alkanes Under Methanogenic Conditions.

Anaerobic alkane degradation via methanogenesis has been intensively studied under mesophilic and thermophilic conditions. While there is a paucity of information on the ability and composition of anaerobic alkane-degrading microbial communities under low temperature conditions. In this study, we investigated the ability of consortium Y15, enriched from Shengli oilfield, to degrade hydrocarbons under different temperature conditions (5-35 °C). The consortium could use hexadecane over a low temperature range (15-30 °C). No growth was detected below 10 °C and above 35 °C, indicating the presence of cold-tolerant species capable of alkane degradation. The preferential degradation of short chain n-alkanes from crude oil was observed by this consortium. The structure and dynamics of the microbial communities were examined using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting and Sanger sequencing of 16S rRNA genes. The core archaeal communities were mainly composed of aceticlastic Methanosaeta spp. Syntrophaceae-related microorganisms were always detected during consecutive transfers and dominated the bacterial communities, sharing 94-96 % sequence similarity with Smithella propionica strain LYP(T). Phylogenetic analysis of Syntrophaceae-related clones in diverse methanogenic alkane-degrading cultures revealed that most of them were clustered into three sublineages. Syntrophaceae clones retrieved from this study were mainly clustered into sublineage I, which may represent psychrotolerant, syntrophic alkane degraders. These results indicate the wide geographic distribution and ecological function of syntrophic alkane degraders.

Preparative separation and purification of four cis-trans isomers of coumaroylspermidine analogs from safflower by high-speed counter-current chromatography.

High-speed counter-current chromatography (HSCCC) was successfully applied for the first time to isolate and purify four cis-trans isomers of coumaroylspermidine analogs from Safflower. HSCCC separation was achieved with a two-phase solvent system composed of chloroform-methanol-water (1:1:1, v/v/v) with the upper phase as the mobile phase. In a single run, a total of 1.3mg of N(1), N(5), N(10)-(E)-tri-p-coumaroylspermidine (EEE), 4.4mg of N(1)(E)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (EZE), 7.2mg of N(1)(Z)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (ZZE), and 11.5mg of N(1),N(5),N(10)-(Z)-tri-p-coumaroylspermidine (ZZZ) were obtained from 100mg of crude sample. High Performance Liquid Chromatography (HPLC) analysis showed that the purities of these four components are 95.5%, 98.1%, 97.5% and 96.2%, respectively. The chemical structures were identified by ESI-MS, (1)H NMR and (13)C NMR.

[Methane-generating potential of coal samples with different maturity].

OBJECTIVE: To evaluate coal bed methane production potential and characterize the in situ microbial communities of coal bed. METHODS: Coal samples were incubated under anaerobic conditions: mimicking coal bed condition, supplementing with methanogenic hydrocarbon degrading consortium, or adding with exogenetic substrate. Methane production was observed over time using gas chromatograph, and the in situ bacterial and archaeal communities were revealed using pyrosequencing. RESULTS: Enrichment incubation revealed that 3 of total 10 coal samples microcosms produced methane; bioaugmentation and substrate addition could enhance methane production of coal sample HF. Hydrogenotrophic Methanoculleus and acetoclastic Methanosaeta dominated the archaeal community of coal sample SL, while the bacterial domain was mainly composed of Firmicutes (54.4%), Proteobacteria (30.9%), uncultured bacteria (10.8%), Caldiserica (1.5%) and Thermotogae (1.3%). CONCLUSION: The methane production potential of coal bed samples with different maturity is different; the in situ coal bed microcosms are likely involved in hydrocarbons degradation and methane production.

Effect of Evn-50 on cell growth and apoptosis in tamoxifen-resistance human breast cancer cell line MCF-7/TAM-R / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of Evn-50 extracted from Vitex negundo on human breast cancer cell line MCF-7 and MCF-7/TAM-R cells in vitro.</p><p><b>METHODS</b>MCF-7 and tamoxifen-resistant MCF-7/TAM-R cells were treated with Evn-50,tamoxifen or combination of Evn-50 and tamoxifen. Cell proliferation inhibition rates were determined by MTT assay. The apoptosis rate and the change of cell cycle were detected by PI staining flow cytometry. Protein expression of phospho-MAPK 44/42 (Thr202/Tyr204),MAPK P44/42, phospho-AKT (Ser473) and AKT were detected with Western blotting.</p><p><b>RESULTS</b>The viability of MCF-7 cells was decreased in combination group [(28.65 ±11.43)%] and Evn-50 group [(53.02 ±15.14)%] compared with TAM group (P<0.01). The cell viability of MCF-7/TAM-R in combination group [(42.11 ±14.30)%] was significantly lower than that in TAM group [(92.18 ±13.16)%] (P<0.01). The cell apoptosis rate was dependent on the time of treatment in all groups,the effects on apoptosis and G2/M phase cells were most prominent at 72 h (P<0.01). Western blotting revealed that protein levels of phosphorylated AKT and p-MAPK44/42 decreased,while the expression of total AKT and MAPK44/42 was stable. In MCF-7/TAM-R cells,the expression of phosphorylation of AKT and MAPK44/42 protein was not changed in Evn-50 or TAM alone group,but significantly inhibited in the combination group at 72 h.</p><p><b>CONCLUSION</b>Evn-50 can inhibit cell growth and induce apoptosis in MCF-7 and MCF-7/TAM-R cells,it can reverse tamoxifen-resistance of MCF-7/TAM-R cells.The mechanisms may be related to the down-regulation of phosphorylated ERK1/2 in MAPK signal pathway and phosphorylated AKT in AKT signal pathway.</p>

[Preventive effect of poria cocos on acute rejection of renal transplantation in rats].

OBJECTIVE: To study the effect of Poria cocos (Pcs) in preventing acute rejection of rats after renal transplantation and its mechanism. METHODS: Rat orthotopic renal transplantation model was performed with Wistar rat as donor and SD rat as donee. All donees were divided into 4 groups, 10 in each group, before transplantation. They were treated respectively with normal saline 5 mL x kg(-1) x d(-1) (A), Pcs 25 mg x kg(-1) x d(-1) (B), Pcs 50 mg x kg(-1) x d(-1) (C) and ciclosporin A (CsA) 5 mg x kg(-1) x d(-1) (D) by intragastric administration. The renal allograft survival time (ST) was recorded, and the serum levels of creatinine (SCr), interleukin-2 (IL-2), gamma-interferon (gamma-IFN), CD4+, CD8+ lymphocytes percentage, CD4+/CD8+ ratio, as well as the pathologic changes were observed one week after transplantation. RESULTS: ST of the renal graft in Groups C and D was significantly longer with pathologic change evidently less than those in Groups A and B (P<0.01), and the ST in Group C was shorter that in Group D (P<0.05). Changes of renal function and urine volume were identified to the pathological change of graft, the initiating time of renal dysfunction was later in Groups C and D than that in Groups A and B. Serum levels of IL-2, IFN-gamma and CD4+ percentage in Group C were significantly lower than those in Groups A and B, but higher than those in Group D respectively (P<0.05 or P<0.01), while CD8+ percentage in Group C was significantly lower than that in Group A (P<0.05), but insignificantly different to that in Groups B and D (P>0.05). CONCLUSION: Pcs shows good dosage-dependent effect in suppressing acute rejection of renal transplantation, but the effect is inferior to that of CsA.