Profile and outcome of receptor conversion in breast cancer metastases: a nation-wide multicenter epidemiological study.

Although receptor status including estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) of the primary breast tumors was related to the prognosis of breast cancer patients, little information is yet available on whether patient management and survival are impacted by receptor conversion in breast cancer metastases. Using data from the nation-wide multicenter clinical epidemiology study of advanced breast cancer in China (NCT03047889), we report the situation of retesting ER, PR and HER2 status for breast cancer metastases and evaluate the patient management and prognostic value of receptor conversion. In total, 3295 patients were analyzed and 1583 (48.0%) patients retesting receptor status for metastasis. Discordance in one or more receptors between the primary and the metastatic biopsy was found in 37.7% of women. Patients who remained hormone receptor (HR) positive in their metastases had similar progression-free survival of first-line and second-line treatment compared to patients with HR conversion (P > .05). In multivariate analysis, patients who showed ER conversion from negative to positive had longer disease-free survival (DFS) than patients who remained negative in their metastases (hazard ratio, 2.05; 95% confidence interval [CI], 1.45-2.90; P < .001). Patients with PR remained positive and had longer DFS than patients with PR conversion from negative to positive (hazard ratio, 0.56; 95% CI, 0.38-0.83; P = .004). Patients with PR conversion have shorter overall survival than patients with PR remained positive or negative (P = .016 and P = .041, respectively). Our findings showed that the receptors' conversions were common in metastatic breast cancer, and the conversion impacted the survival.

Molecularly imprinted polymers on a silica surface for the adsorption of tobacco-specific nitrosamines in mainstream cigarette smoke.

Tobacco-specific nitrosamines are one of the most important groups of carcinogens in tobacco products. Using adsorbents as filter additives is an effective way to reduce tobacco-specific nitrosamines in cigarette smoke. Molecularly imprinted polymers (MIPs) using nicotinamide as template were grafted on the silica gel surface to obtain MIP@SiO2 and employed as filter additives to absorb tobacco-specific nitrosamines in mainstream cigarette smoke. Four milligrams of MIP@SiO2 per cigarette was added to the interface between filter and tobacco rod to prepare a binary filter system. The mainstream smoke was collected on an industry-standard Cambridge filter pad and extracted with ammonium acetate aqueous solution before analysis. Compared to the cigarette smoke of the control group, the levels of tobacco-specific nitrosamines with silica gel and with MIP@SiO2 were both reduced, and the adsorption rates of N-nitrosonornicotine, N-nitrosoanabasine, N-nitrosoanatabine, and 4-(methylnitrosamino)-1-(3-pyridine)-1-butanone with silica gel and with MIP@SiO2 were 20.76, 15.32, 18.79, and 18.01%, and 41.33, 34.04, 37.86, and 35.53%, respectively. Furthermore the content of total particle materials in cigarette smoke with silica gel was decreased evidently but showed no observable change with MIP@SiO2 . It indicated MIP@SiO2 could selectively reduce tobacco-specific nitrosamines in the mainstream cigarette smoke with no change to the cigarette flavor.

Effect of arsenite-oxidizing bacterium B. laterosporus on arsenite toxicity and arsenic translocation in rice seedlings.

Arsenite [As (III)] oxidation can be accelerated by bacterial catalysis, but the effects of the accelerated oxidation on arsenic toxicity and translocation in rice plants are poorly understood. Herein we investigated how an arsenite-oxidizing bacterium, namely Brevibacillus laterosporus, influences As (III) toxicity and translocation in rice plants. Rice seedlings of four cultivars, namely Guangyou Ming 118 (GM), Teyou Hang II (TH), Shanyou 63 (SY) and Minghui 63 (MH), inoculated with or without the bacterium were grown hydroponically with As (III) to investigate its effects on arsenic toxicity and translocation in the plants. Percentages of As (III) oxidation in the solutions with the bacterium (100%) were all significantly higher than those without (30-72%). The addition of the bacterium significantly decreased As (III) concentrations in SY root, GM root and shoot, while increased the As (III) concentrations in the shoot of SY, MH and TH and in the root of MH. Furthermore, the As (III) concentrations in the root and shoot of SY were both the lowest among the treatments with the bacterium. On the other hand, its addition significantly alleviated the As (III) toxicity on four rice cultivars. Among the treatments amended with B. laterosporus, the bacterium showed the best remediation on SY seedlings, with respect to the subdued As (III) toxicity and decreased As (III) concentration in its roots. These results indicated that As (III) oxidation accelerated by B. laterosporus could be an effective method to alleviate As (III) toxicity on rice seedlings.

[Clinical features and prognosis in 104 colorectal cancer patients with bone metastases].

OBJECTIVE: To investigate the clinical features and prognosis of bone metastases in colorectal cancer patients. METHODS: The clinical data of 104 cases of colorectal cancer with bone metastasis were collected and retrospectively analyzed. RESULTS: Among all the 104 patients included, 45 (43.3%) patients had multiple bone metastases, and 59 (56.7%) patients had single bone metastasis. Pelvis (46.1%) was the most common site, followed by thoracic vertebrae (41.3%), lumbar vertebrae (40.4%), sacral vertebrae (29.8%) and ribs (29.8%). One hundred and two patients (98.1%) were complicated with other organ metastases. The median time from colorectal cancer diagnosis to bone metastasis was 16 months, and the median time from bone metastasis to first skeletal-related events (SREs) was 1 month. The most common skeletal-related events (SREs) were the need for radiotherapy (44.2%), severe bone pain (15.4%) and pathologic fracture (9.6%). The median survival time of patients with bone metastases was 10.0 months, and 8.5 months for patients with SREs. ECOG score, systemic chemotherapy and bisphosphonate therapy were prognostic factors by univariate analysis (all P < 0.05). ECOG score and systemic chemotherapy were independent prognostic factors by Cox multivariate analysis. CONCLUSIONS: Bone metastasis in colorectal cancer patients has a poor prognosis and the use of chemotherapy and bisphosphonates may have a benefit for their survival.

[The analysis and identification of fritillaria cirrhosa by Raman spectra].

Laser Raman spectrum technology was used to distinguish fritillaria cirrhosa from its adulterants rapidly and accurately. The study is based on that different traditional Chinese medicine contains different chemical compositions, and the differences could be displayed in Raman spectra. The Raman spectra of fritillaria cirrhosas shows that several characteristic strong peaks could be found at 442, 480, 863, 941, 1 083, 1 129, 1 342, 1 463 and 2 910 cm(-1), and a few obvious peaks appear at 111, 302, 360, 409, 527, 579, 618, 718, 767, 1 052, 1 083, 1 207 and 1 261 cm(-1). According to the Raman spectra, a Raman fingerprint of fritillaria cirrhosa was set up, which could be used to distinguish its adulterants. This analysis could be used in identifying fritillaria cirrhosa rapidly, accurately and nondestructively.

Perception of recurrent risk versus objective measured risk of ischemic stroke in first-ever stroke patients from a rural area in China: A cross-sectional study.

OBJECTIVE: Risk perception is critical to the formation of individual health prevention behaviors. A long-term accurate perception of stroke recurrent risks is imperative for stroke secondary prevention. This study aims to explore the level of recurrence risk perceptions and the influential factors of inaccuracy between perceived and objective risk in first-ever ischemic stroke patients from a rural area. METHODS: From May to November 2020, 284 first-ever ischemic stroke patients were conveniently recruited in a rural area of Henan Province, China. Perceived risk was measured based on self-reported using a numerical rating scale, whereas the objective risk was measured by the Essen Stroke Risk Score. Patients' perceived risk was compared with their objective risk and categorized as "Accurate," "Underestimated," and "Overestimated." The influencing factors of inaccuracy were further evaluated using multivariate regression analyses. RESULTS: 46% of the participants underestimated their stroke risk, while 15.9% overestimated their risks. Patients who were younger (≤65 years), didn't worry about recurrent stroke, and had a low actual recurrent risk were more likely to underestimate their recurrent risk. Patients who were employed, had lower independence, and had greater anxiety were more likely to overestimate their recurrent risk. CONCLUSIONS: The majority of participants were unable to accurately perceive their own risk of stroke recurrence. Patients' age, working status, worry about recurrent stroke, actual recurrent risk, level of dependence, and anxiety played a role in perception inaccuracy. PRACTICE IMPLICATIONS: The findings could help healthcare providers gain a better understanding of the level and accuracy of recurrence risk perceptions among first-ever stroke patients in the rural area. Future counseling on the perceived risk of stroke recurrence and individual objective risk assessment could be conducted to help patients better understand their risk of recurrence. Individualized risk communication and multidisciplinary teamwork can be developed to improve the accuracy of recurrence risk perceptions and health behaviors.

Physio-psycho-social interaction mechanism in dyadic health of young and middle-aged stroke survivors and their spousal caregivers: a longitudinal observational study protocol.

INTRODUCTION: In recent years, stroke has become more common among young people. Stroke not only has a profound impact on patients' health but also incurs stress and health threats to their caregivers, especially spousal caregivers. Moreover, the health of stroke survivors and their caregivers is interdependent. To our knowledge, no study has explored dyadic health of young and middle-aged stroke survivors and their spousal caregivers from physiological, psychological and social perspectives. Therefore, this proposed study aims to explore the mechanism of how physiological, psychological and social factors affect dyadic health of young and middle-aged stroke survivors and their spousal caregivers. The findings of this study will provide implications for developing interventions to improve dyadic health of this growing population. METHODS AND ANALYSES: We will collect data from 57 dyads of young and middle-aged stroke survivors and their spousal caregivers during hospitalisation and at 1, 3, 6, 9 and 12 months after discharge. Questionnaires will be used to collect participants' demographic information, stress, depression, anxiety, benefit finding, social support, mutuality and quality of life. The following physiological reactions will be collected at baseline, including interleukin 6, tumour necrosis factor-alpha and salivary cortisol. ETHICS AND DISSEMINATION: The study was approved by the ethics review committee of life sciences of Zhengzhou University (No. ZZUIRB2020-53). Prior to being enrolled in the study, participants will be given full and detailed information about the possible risks involved, the informed consent process, confidentiality, the study procedure and secure data storage. Participants will be guaranteed that they can withdraw from the study at any time without providing a reason or leading to any consequences. Both oral and written informed consent will be obtained from all participants. The findings of this proposed study will be disseminated through peer-reviewed journals and academic conferences.

[HPLC determination of the related substances in erdosteine].

An HPLC method was established for the determination of the related substance in erdosteine. Waters ODS-SunFire (250 mm x 4.6 mm ID, 5 microm) column was used, the mobile phase was composed of methanol-acetonitrile-0.01 mol x L(-1) citric acid (20:4:76, the pH value was adjusted by triethylamine to 2.5). The flow rate was 1 mL x min(-1). The detection wavelength was 254 nm. The related substances in the sample of erdosteine taken were calculated by self control with or without the response factor of impurity relative to that of erdosteine. Under the chromatographic condition developed, the impurities in erdosteine were isolated well. The detection limit was 0.2 microg x mL(-1) (signal/noise = 3) by principal component calculated. The method can be adopted to control the related substances in erdosteine.

Thrombolysis after cardiopulmonary resuscitation in myocardial infarction with abdominal pain as the first presentation: A case report.

RATIONALE: Thrombolysis after cardiopulmonary resuscitation in patients with acute ST-segment elevation myocardial infarction (STEMI) is controversial. This case report describes a successful thrombolysis after resuscitation in delayed-diagnosis STEMI. PATIENT CONCERNS: A 58-year-old man presented with acute abdominal pain as the first symptom of a subsequent STEMI diagnosis. When he returned to the clinic after having been assisted with abdominal pain relief, he suffered a sudden cardiac arrest. Cardiopulmonary resuscitation was performed immediately, and thrombolysis was carried out for his anterior STEMI. He was successfully resuscitated in a short period of time. DIAGNOSIS: The patient was diagnosed with acute and extensive anterior STEMI. The D-dimer level was normal, and pericardial effusion was ruled out. INTERVENTIONS: After successful resuscitation, the patient received half-dose alteplase thrombolytic therapy. After a few days, the patient was transferred to a general ward. Coronary angiography revealed unobstructed flow in the left anterior descending artery. OUTCOMES: The ST segment of the patient gradually declined after thrombolytic therapy, and the myocardial injury marker levels increased. A small amount of pleural fluid in the lungs and pulmonary infection were observed. With effective diuretic, anti-infective, and other treatments, the patient's condition gradually improved, the ventilator was removed, and vasoactive drugs were successfully discontinued. Coronary angiography revealed that the flow of the culprit artery was unobstructed, and a drug-coated balloon was implanted. No wall motion abnormalities were detected on echocardiography, and the patient recovered well. CONCLUSIONS: In patients with abdominal pain as the first presentation, a simple initial electrocardiogram may help reduce the risk of missed STEMI diagnosis. Thrombolysis after successful resuscitation is an effective treatment for these patients. However, the effects of thrombolysis after resuscitation remain unclear. The point of dispute lies in the effectiveness and safety of thrombolysis (primarily for bleeding). Prompt thrombolysis would lead to a better prognosis if spontaneous circulation can be restored within 10 minutes.

Expression dynamics of periodic transcripts during cancer cell cycle progression and their correlation with anticancer drug sensitivity.

BACKGROUND: The cell cycle is at the center of cellular activities and is orchestrated by complex regulatory mechanisms, among which transcriptional regulation is one of the most important components. Alternative splicing dramatically expands the regulatory network by producing transcript isoforms of genes to exquisitely control the cell cycle. However, the patterns of transcript isoform expression in the cell cycle are unclear. Therapies targeting cell cycle checkpoints are commonly used as anticancer therapies, but none of them have been designed or evaluated at the alternative splicing transcript level. The utility of these transcripts as markers of cell cycle-related drug sensitivity is still unknown, and studies on the expression patterns of cell cycle-targeting drug-related transcripts are also rare. METHODS: To explore alternative splicing patterns during cell cycle progression, we performed sequential transcriptomic assays following cell cycle synchronization in colon cancer HCT116 and breast cancer MDA-MB-231 cell lines, using flow cytometry and reference cell cycle transcripts to confirm the cell cycle phases of samples, and we developed a new algorithm to describe the periodic patterns of transcripts fluctuating during the cell cycle. Genomics of Drug Sensitivity in Cancer (GDSC) drug sensitivity datasets and Cancer Cell Line Encyclopedia (CCLE) transcript datasets were used to assess the correlation of genes and their transcript isoforms with drug sensitivity. We identified transcripts associated with typical drugs targeting cell cycle by determining correlation coefficients. Cytotoxicity assays were used to confirm the effect of ENST00000257904 against cyclin dependent kinase 4/6 (CDK4/6) inhibitors. Finally, alternative splicing transcripts associated with mitotic (M) phase arrest were analyzed using an RNA synthesis inhibition assay and transcriptome analysis. RESULTS: We established high-resolution transcriptome datasets of synchronized cell cycle samples from colon cancer HCT116 and breast cancer MDA-MB-231 cells. The results of the cell cycle assessment showed that 43,326, 41,578 and 29,244 transcripts were found to be periodically expressed in HeLa, HCT116 and MDA-MB-231 cells, respectively, among which 1280 transcripts showed this expression pattern in all three cancer cell lines. Drug sensitivity assessments showed that a large number of these transcripts displayed a higher correlation with drug sensitivity than their corresponding genes. Cell cycle-related drug screening showed that the level of the CDK4 transcript ENST00000547281 was more significantly associated with the resistance of cells to CDK4/6 inhibitors than the level of the CDK4 reference transcript ENST00000257904. The transcriptional inhibition assay following M phase arrest further confirmed the M-phase-specific expression of the splicing transcripts. Combined with the cell cycle-related drug screening, the results also showed that a set of periodic transcripts, for example, ENST00000314392 (a dolichyl-phosphate mannosyltransferase polypeptide 2 isoform transcript), was more associated with drug sensitivity than the levels of their corresponding gene transcripts. CONCLUSIONS: In summary, we identified a panel of cell cycle-related periodic transcripts and found that the levels of transcripts of drug target genes showed different values for predicting drug sensitivity, providing novel insights into alternative splicing-related drug development and evaluation.

Unmet care needs of community-dwelling stroke survivors: a systematic review of quantitative studies.

OBJECTIVES: Understanding the unmet needs of community-dwelling stroke survivors is essential for further intervention. This systematic review was performed to summarise their unmet needs from a quantitative viewpoint. DESIGN: Systematic review using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. DATA SOURCES: A comprehensive search of six databases was conducted from inception to February 2020: PubMed, EMBASE, CINAHL, PsycINFO, SCOPUS and CBM. The methodological quality of the studies was assessed. Unmet needs were categorised, and a pooled analysis of the main outcomes was conducted. ELIGIBILITY CRITERIA FOR SELECTING STUDIES: We included quantitative studies focused on the unmet needs of stroke survivors who live at homes rather than in any other institutionalised organisation. RESULTS: In total, 32 of 2660 studies were included, and 1980 unmet needs were identified. The prevalence of patients with unmet needs ranged from 15.08% to 97.59%, with a median of 67.20%; the median number of unmet needs per patient ranged from 2 to 8 (0-31). The prevalence of unmet needs was high at 6 months post-stroke (62.14%) and 2 years post-stroke (81.37%). After categorisation, the main concerns among these patients were revealed to be information support, physical function and mental health; a few studies reported unmet needs related to leisure exercise, return to work and so on. Additionally, differences in the measurement tools used across studies affect what unmet needs participants report. CONCLUSIONS: Sufficient, accurate, individualised and dynamic information support is a priority among community-dwelling stroke survivors. Physical function and mental health are also the most significant concerns for re-achieving social participation. It is essential to design and disseminate standard, effective and time-saving tools to assess unmet needs. TRIAL REGISTRATION NUMBER: CRD42018112181.

Comparisons of Treatment for HER2-Positive Breast Cancer between Chinese and International Practice: A Nationwide Multicenter Epidemiological Study from China.

OBJECTIVE: To better understand the status of medical treatment for human epidermal growth factor receptor 2 (HER2)-positive breast cancer and the differences between the Chinese and the international clinical practice. METHODS: This was a retrospective, nationwide, multicenter, epidemiological study of advanced breast cancer patients from China. Between January 01, 2012, and December 31, 2014, a total of 3649 patients, covering 7 geographic regions and 21 institutions, participated in this series of studies. HER2-positive breast cancer was selected among the group and adopted into this study. In comparison, we summarized the demographics and clinical characteristics of HER2-positive breast cancer from the Surveillance, Epidemiology, and End Results (SEER) database. RESULTS: A total of 918 patients diagnosed as HER2-positive breast cancer patients were included. The median age at diagnosis was 46 years (ranging, 23 to 78) with a single-peak incidence. The proportions of stages II-IV at diagnosis and distance metastasis in viscera were more than half of the participants. In comparison, the prevalence of estrogen or progesterone receptor-positive expression and luminalB subtype was relatively lower than that of the United States. The receipt of chemotherapy was fairly higher, while the usage of targeted therapy was seriously insufficient. Tumor size was in significantly positive associations with the duration of targeted therapy (Kendall's correlation coefficient = 0.3, P < 0.0001), while no prohibitive variables among clinical characteristics were detected. CONCLUSION: Our study suggested that HER2-positive breast cancer patients were characterized as a younger trend, a lower prevalence of hormonal receptor (HR)-positive expression, and less accessible to anti-HER2 targeted therapy with insufficient duration over the past few years in China. Concerted efforts should be exerted for promising survival benefits in the future. The trial registration number is https://clinicaltrials.gov/ct2/show/NCT03047889.

Diversity and Bioactivity of Endophytes From Angelica sinensis in China.

Plant seeds are not merely reproductive organs, they are also carriers of microorganism, particularly, inherent and non-invasive characteristic endophytes in host plant. Therefore, in this study, the endophytic diversity of Angelica seeds was studied and compared with endophytes isolated from healthy leaves, stems, roots, and seeds of A. sinensis using 20 different media. The metabolites of endophytic strains were evaluated with six different methods for their antioxidant activity and the paper disc diffusion method for antimicrobial activities. As a result, 226 endophytes were isolated. Compared with the biodiversity and abundance of uncultured fungi from Angelica seed, the result showed that the most frequent endophytic fungi were Alternaria sp. as seen in artificial media; moreover, compared with artificial media, the pathogenic fungi, including Fusarium sp. and Pseudallescheria sp., were not found from the Angelica seed, the results suggested it may not be inherent endophytes in plants. In addition, bacteria from seven phyla were identified by high-throughput sequencing, while five phyla of endophytic bacteria were not isolated on artificial media including Proteobacteria, Actinobacteria, Bacteroidetes, Microgenomates, and Saccharibacteria. Furthermore, the sample JH-4 mycelium displayed the best antioxidant activity, and the active constituent may be a flavonoid as determined by total phenol and flavonoid content. Moreover, YH-12-1 mycelium had strong inhibitory activity against the five tested strains and the minimum inhibitory concentration (MIC) against Pseudomonas aeruginosa and Streptococcus pneumoniae was found to be 25 µg/mL. Our results confirm that plant endophytes are rich in biodiversity and contain important resource of many uncultured microorganisms.

Ezetimibe Attenuates Oxidative Stress and Neuroinflammation via the AMPK/Nrf2/TXNIP Pathway after MCAO in Rats.

Oxidative stress and neuroinflammation play essential roles in ischemic stroke-induced brain injury. Previous studies have reported that Ezetimibe (Eze) exerts antioxidative stress and anti-inflammatory properties in hepatocytes. In the present study, we investigated the effects of Eze on oxidative stress and neuroinflammation in a rat middle cerebral artery occlusion (MCAO) model. One hundred and ninety-eight male Sprague-Dawley rats were used. Animals assigned to MCAO were given either Eze or its control. To explore the downstream signaling of Eze, the following interventions were given: AMPK inhibitor dorsomorphin and nuclear factor erythroid 2-related factor 2 (Nrf2) siRNA. Intranasal administration of Eze, 1 h post-MCAO, further increased the endogenous p-AMPK expression, reducing brain infarction, neurologic deficits, neutrophil infiltration, microglia/macrophage activation, number of dihydroethidium- (DHE-) positive cells, and malonaldehyde (MDA) levels. Specifically, treatment with Eze increased the expression of p-AMPK, Nrf2, and HO-1; Romo-1, thioredoxin-interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3), Cleaved Caspase-1, and IL-1ß were reduced. Dorsomorphin and Nrf2 siRNA reversed the protective effects of Eze. In summary, Eze decreases oxidative stress and subsequent neuroinflammation via activation of the AMPK/Nrf2/TXNIP pathway after MCAO in rats. Therefore, Eze may be a potential therapeutic approach for ischemic stroke patients.

[Effects of defoliation on the allocation of non-structural carbohydrates in roots of Fraxinus mandshurica seedlings.] / åŽ»å¶å¯¹æ°´æ›²æŸ³è‹—æœ¨æ ¹ç³»éžç»“æž„æ€§ç¢³æ°´åŒ–åˆç‰©åˆ†é çš„å½±å“.

Global climate changes would lead to outbreaks of leaf-feeding insects. Leaf loss could reduce photosynthate production, with consequences on non-structural carbohydrates (NSC) storage and allocation in trees. In this study, the responses of NSC and its compartment concentrations in tap-, coarse- and the first to fifth order fine roots of 2-year-old seedlings of Fraxinus mandshurica to defoliation (40% loss of leaf area) were measured from June to October. The results showed that NSC and its compartment concentrations in roots exhibited distinct seasonal dynamics in both control and defoliation treatments. Following defoliation, NSC concentration decreased in tap- and coarse roots by 3.8% and 30.7%, respectively, while increased in the first five order roots by 1.2%-23.5%, to which starch contributed majorly for each root compartment. Soluble sugar concentration was enhanced by defoliation in tap- and coarse roots by 7.1% and 62.3%, respectively, but decreased in the first to fifth order roots by 2.7%-42.8%. Defoliation had different influences on starch and soluble sugar, with positive effects on the ratio of soluble sugar to starch concentrations in tap- and coarse roots but negative effects on the first to fifth order roots. Overall, defoliation decreased photosynthate production in leaves, leading to the remobilization of starch in tap- and coarse roots and the transportation as soluble sugar to fine roots, as well as the following storage in these roots, which would facilitate the resistance of fine roots to the low temperature in winter.

A novel enzyme-assisted approach for efficient extraction of Z-ligustilide from Angelica sinensis plants.

Endophytes coexist with plants, in part, due to cellulase that allow saccharification of plant cell walls. The cellulase enzymes found in naturally occurring endophytes may exhibit stronger activity and more specificity than commercially available cellulase for enzyme-assisted extraction of compounds from medicinal plant materials. In order to identify endophytes with high cellulase activity, we screened endophytes taken from different parts of Angelica sinensis using the Congo red staining method. We identified three strains with higher cellulase activity. Of the 3 strains identified, No.Lut1201 increased the yield of extracted Z-ligustilide 2 fold compared to commercially available cellulase (Ningxia Sunson) using a cellulase-assisted extraction method and traditional extraction methods. Scanning electron microscopy clearly demonstrated that the cellulase extracted from endophytes enhance cell wall polysaccharide degradation as well as Z-ligustilide extraction from Radix Angelica sinensis (RAS). The current study provides a new method and ideas of using cellulase of endophytes for improving the extraction of compounds from medicinal plants.

Expression and Purification of Delta Sleep-Inducing Peptide Fused with Protein Transduction Domain and Human Serum Albumin in Pichia pastoris.

BACKGROUND: Sleep is a natural part of every individual's life. Delta sleep-inducing peptide (DSIP) is a nonapeptide that could promote sleep through the induction of slow wave sleep. However, little is known about the pharmacological effect of DSIP on insomnia. OBJECTIVES: The main objective of this study was to analyze the pharmacological effect of DSIP on insomnia. METHODS: We designed a fusion protein containing N-terminal TAT-based transduction domain followed by human serum albumin and DSIP and designated this protein as PHD fusion protein. The PHD fusion protein were expressed in Pichia pastoris and purified. Mice were administered single subcutaneous injections three concentrations of PHD fusion protein (0.5, 1, 2 mg/kg), and the pharmacological activity of PHD fusion protein was studied using classic pentobarbitalinduced sleep test. RESULTS: We expressed the PHD fusion protein in P. pastoris; furthermore, the PHD fused protein was purified to near homogeneity by DEAE Sepharose FF, Phenyl Sepharose HP and Blue Sepharose 6 FF. Our result showed that the increase of pentobarbital-induced hypnotic effect characterized by reducing sleep latency and prolonged sleep duration was observed for increasing concentrations of PHD fusion protein (P<0.05); moreover, different dose of PHD fusion protein could induce the mice to re-sleep in a dose-dependent manner, whereas higher doses of PHD fusion protein (1.0, 2.0 mg/kg) significantly increased the rate of sleep re-onset compared with the vehicle group of mice (P<0.05). CONCLUSION: PHD fusion protein increased the hypnotic effects of pentobarbital by reducing sleep latency and prolonged sleep duration. The present study suggested PHD fusion protein could be a new drug candidate for insomnia.

Effect of X-ray irradiation on hepatocarcinoma cells and erythrocytes in salvaged blood.

The broad clinical acceptance of intraoperative blood salvage and its applications in cancer surgery remain controversial. Until now, a method that can safely eliminate cancer cells while preserving erythrocytes does not exist. Here, we investigated whether X-ray generated from linear accelerator irradiation at a certain dose can kill hepatocarcinoma cells while preserving erythrocytes. HepG2, SK-Hep1 or Huh7 cells were mixed into the aliquots of erythrocytes obtained from healthy volunteers. After the mixed cells were exposed to 30 Gy and 50 Gy X-rays irradiation, the viability, clonogenicity, DNA synthesis and tumorigenicity of the tumor cells were determined by the MTT assay, plate colony formation, 5-ethynyl-2'-deoxyuridine incorporation, and subcutaneous xenograft implantation into immunocompromised mice. The ATP, 2,3-DPG, free Hb, osmotic fragility, blood gas variables in erythrocytes and morphology of erythrocytes at 0 h, 12 h, 24 h, 48 h, 72 h after irradiation were analyzed. X-ray irradiation at 30 Gy effectively inhibited the viability, proliferation, and tumorigenicity of HepG2, SK-Hep1 and Huh7 cells without noticeably damaging the ability of oxygen-carrying, membrane integrity and morphology of erythrocytes. Theses results suggest that X-ray at 30 Gy irradiation might be safe to eliminate hepatocarcinoma cells while preserving erythrocytes in salvaged blood.

EphrinB-EphB Signaling Induces Hyperalgesia through ERK5/CREB Pathway in Rats.

BACKGROUND: There are numerous studies implicating that EphB receptors and ephrinB ligands play important roles in modulating the transduction of spinal nociceptive information. EphrinB-EphB signaling may contribute to hyperalgesia via various kinds of downstream molecules, the mechanisms of which have not been completely understood. OBJECTIVE: The aim of the present study was to identify whether ephrinB-EphB signaling could contribute to hyperalgesia through ERK5/CREB pathway. STUDY DESIGN: Controlled animal study. SETTING: University laboratory. METHODS: This study attempted to detect the changes of pain behaviors and the protein level of p-ERK5 and p-CREB by activating EphB receptors in the spinal cord of rats. To further confirm our hypothesis, we designed LV-siRNA for knockdown of spinal ERK5. When ERK5 was inhibited, we recorded the changes of spinal p-CREB expression and the pain behaviors of rats after activating EphB receptors. We also confirmed this conclusion in rat CCI model. Statistical analyses were performed using GraphPad Prism 5. RESULTS: Intrathecal injection of ephrinB2-Fc in rats evoked thermal hyperalgesia and mechanical allodynia, along with activation of ERK5 and CREB in the spinal cord. Knockdown of ERK5 inhibited ephrinB2-Fc-induced CREB activation and hyperalgesia. Blocking EphB receptors prevented CCI-induced neuropathic pain and spinal ERK5/CREB activation. LIMITATIONS: More underlying mechanisms that underlie the relationship between ephrinB-EphB signaling and ERK5/CREB pathway will need to be explored in future studies. CONCLUSIONS: Our study suggests that ERK5/CREB pathway plays important roles in the transduction of nociceptive information associated with ephrinB-EphB signaling. This study provides further understanding of the downstream mechanisms of ephrinB-EphB signaling and helps to explore new targets for treating pathological pain.

[Effects of elevated N availability on anatomy, morphology and mycorrhizal colonization of fine roots: A review.] / æ°®æœ‰æ•ˆæ€§å¢žåŠ å¯¹ç»†æ ¹è§£å‰–ã€å½¢æ€ç‰¹å¾å’ŒèŒæ ¹ä¾µæŸ“çš„å½±å“.

Increase of nitrogen (N) availability can greatly affect the structure and function of forest ecosystems. Fine root is critical to belowground ecological processes, thus its response to elevated N availability has become a focus of ecological researches. This article reviewed the trend and under-lying mechanism of fine root response to elevated N availability, including root anatomy, morphology and mycorrhizal colonization. We found that cross-sectional areas of root stele and xylem and the diameter, number and wall thickness of xylem vessel (tracheid) all increased, however, cortex thickness, the number of cortical cell layer, diameter of cortical cell and number of passage cell in exodermis decreased with higher N availability. The response of root anatomy was closely related to plant hormones. In addition, mycorrhiza colonization rate decreased after N fertilization, while specific root length (root length per unit root mass) generally decreased in arbuscular mycorrhizal species, while increased in ectomycorrhizal species. By contrast, the root diameter and tissue density varied widely among species under higher N status. These findings based on individual roots and species provided deeper understanding of carbon and nutrient cycles in terrestrial ecosystems. In addition, we discussed some knowledge gaps and proposed several research outlooks for guiding future researches.