Activation of RSK2 upregulates SOX8 to promote methotrexate resistance in gestational trophoblastic neoplasia.

Resistance to chemotherapy is frequently driven by aberrantly activated kinases in cancer. Herein, we characterized the global phosphoproteomic alterations associated with methotrexate (MTX) resistance in gestational trophoblastic neoplastic (GTN) cells. A total of 1111 phosphosites on 713 proteins were significantly changed, with highly elevated Ribosomal S6 Kinase 2 (RSK2) phosphorylation (pS227) observed in MTX-resistant GTN cells. Activation of RSK2 promoted cell proliferation and survival after MTX treatment in GTN cell models. Interestingly, RSK2 might play an important role in the regulation of reactive oxygen species (ROS) homeostasis, as manipulation of RSK2 activation affected ROS accumulation and SOX8 expression in GTN cells. In addition, overexpression of SOX8 partly rescued cell proliferation and survival in RSK2-depleted MTX-resistant GTN cells, suggesting that SOX8 might serve as a downstream effector of RSK2 to promote MTX resistance in GTN cells. Highly activated RSK2/SOX8 signaling was observed in MTX-resistant GTN specimens. Further, the RSK2 inhibitor BIX02565 effectively reduced SOX8 expression, induced ROS accumulation, and enhanced MTX-induced cytotoxicity in vitro and in vivo. Collectively, our findings suggested that RSK2 activation could promote MTX resistance via upregulating SOX8 and attenuating MTX-induced ROS in GTN cells, which may help to develop experimental therapeutics to treat MTX-resistant GTN.

Fibroblast growth factor 21 enhances angiogenesis and wound healing of human brain microvascular endothelial cells by activating PPARγ.

Angiogenesis of brain microvascular endothelial cells (BMECs) is required in the functional restoration of brain injury, such as traumatic brain injury (TBI) and ischemic stroke. Fibroblast growth factor 21 (FGF21) is an angiogenic molecule that functions through the formation of the FGF21/FGFR1/ß-klotho complex but does not cause carcinogenic events. The current study was to determine whether recombinant human FGF21 (rhFGF21) could promote angiogenesis and scratch wound healing of human brain microvascular endothelial cells (HBMECs) and the possible underlying mechanism. rhFGF21 promoted angiogenesis and migration of HBMECs. The FGFR1 inhibitor PD173074 was applied to demonstrate that rhFGF21 functions through the formation of FGF21/FGFR1/ß-klotho complexes. In addition, the specific PPARγ inhibitor GW9662 and PPARγ activator rosiglitazone were applied to determine that the role of rhFGF21 in increasing angiogenesis is through the PPARγ pathway. In addition, we revealed that the effect of rhFGF21 acts partially through upregulating eNOS expression. In conclusion, our study provides novel evidence that rhFGF21 can enhance the angiogenesis and migration of HBMECs through the formation of the FGF21/FGFR1/ß-klotho complex via PPARγ activation and eNOS upregulation, indicating that FGF21 is a potential therapeutic angiogenic agent for the treatment of human brain injury.

An endoplasmic reticulum protein, Nogo-B, facilitates alcoholic liver disease through regulation of kupffer cell polarization.

Nogo-B (Reticulon 4B) is an endoplasmic reticulum (ER) resident protein that regulates ER structure and function. Because ER stress is known to induce M2 macrophage polarization, we examined whether Nogo-B regulates M1/M2 polarization of Kupffer cells and alters the pathogenesis of alcoholic liver disease (ALD). M1 and M2 phenotypes were assessed in relation to Nogo-B expression and disease severity in liver specimens from ALD patients (NCT01875211). Liver specimens from wild-type (WT) and Nogo-B knockout (KO) mice fed a control or Lieber-DeCarli ethanol liquid diet (5% ethanol) for 6 weeks were analyzed for liver injury and steatosis. Kupffer cells isolated from WT and Nogo-B KO mice were assessed for M1 and M2 activation. A significant positive correlation was observed between Nogo-B positive Kupffer cells and disease severity in ALD patients (n = 30, r = 0.66, P = 0.048). Furthermore, Nogo-B-positive Kupffer cells were correlated with M1 activation (inducible nitric oxide synthase) (r = 0.50, P = 0.05) and negatively with markers of M2 status (CD163) (r = -0.48, P = 0.07) in these patients. WT mice exhibited significantly increased liver injury (P < 0.05) and higher hepatic triglyceride levels (P < 0.01) compared with Nogo-B KO mice in response to chronic ethanol feeding. Nogo-B in Kupffer cells promoted M1 polarization, whereas absence of Nogo-B increased ER stress and M2 polarization in Kupffer cells. CONCLUSION: Nogo-B is permissive of M1 polarization of Kupffer cells, thereby accentuating liver injury in ALD in humans and mice. Nogo-B in Kupffer cells may represent a new therapeutic target for ALD. (Hepatology 2017;65:1720-1734).

IL-3 and CTLA4 gene polymorphisms may influence the tacrolimus dose requirement in Chinese kidney transplant recipients.

The highly variable pharmacokinetics and narrow therapeutic window of tacrolimus (TAC) has hampered its clinical use. Genetic polymorphisms may contribute to the variable response, but the evidence is not compelling, and the explanation is unclear. In this study we attempted to find previously unknown genetic factors that may influence the TAC dose requirements. The association of 105 pathway-related single nucleotide polymorphisms (SNPs) with TAC dose-adjusted concentrations (C0/D) was examined at 7, 30 and 90 d post-operation in 382 Chinese kidney transplant recipients. In CYP3A5 non-expressers, the patients carrying the IL-3 rs181781 AA genotype showed a significantly higher TAC logC0/D than those with the AG genotype at 30 and 90 d post-operation (AA vs AG, 2.21±0.06 vs 2.01±0.03, P=0.004; and 2.17±0.06 vs 2.03±0.03, P=0.033, respectively), and than those with the GG genotype at 30 d (AA vs GG, 2.21±0.06 vs 2.04±0.03, P =0.011). At 30 d, the TAC logC0/D in the grouped AG+GG genotypes of CTLA4 rs4553808 was significantly lower than that in the AA genotype (P =0.041) in CYP3A5 expressers, but it was higher (P=0.008) in the non-expressers. We further validated the influence of CYP3A5 rs776746, CYP3A4 rs2242480 and rs4646437 on the TAC C0/D; other candidate SNPs were not associated with the differences in TAC C0/D. In conclusion, genetic polymorphisms in the immune genes IL-3 rs181781 and CTLA4 rs4553808 may influence the TAC C0/D. They may, together with CYP3A5 rs776746, CYP3A4 rs2242480 and rs4646437, contribute to the variation in TAC dose requirements. When conducting individualized therapy with tacrolimus, these genetic factors should be taken into account.

[Risk factors for the formation of aberrant artery collaterals in the uterus with scar in pregnancy womenunderwent cesarean].

OBJECTIVE: To evaluate the prevalence and risk factors for the formation of aberrant artery collaterals in the uterus during uterine artery embolization (UAE).
 METHODS: The data of 144 women with scar in the uterus due to cesarean were retrospectively analyzed. They underwent UAE in the period of 2009-2014 and were divided into two groups according to a standard with or without the aberrant artery collaterals in the uterus. The risk factors were analyzed.
 RESULTS: Aberrant artery collaterals were found in thirty-four patients. According to multiple logistic regression analysis, the presence of placenta previa (RR=78.556, 95% CI: 2.869-2 150.651, P=0.010), pelvic inflammatory disease (RR=6.633, 95% CI: 1.595-27.592, P=0.009), pregnancy complications (RR=7.264, 95% CI: 1.622-32.531, P=0.010), abortions (RR=18.381, 95% CI: 1.683-200.752, P=0.017) and uterine fibroids or adenomyosis (RR=12.580, 95% CI: 1.004-157.550, P=0.050) were the factors for the presence of aberrant artery collaterals.
 CONCLUSION: Aberrant artery collaterals were more frequent in patients with pelvic inflammatory disease, pregnancy complications, abortions and uterine fibroids or adenomyosis.

[Diagnosis and treatment of vascular complications of external iliac arteries after kidney transplantation: a report of 6 cases].

OBJECTIVE: To explore the characteristics of external iliac artery vascular complications after renal transplantation and the diagnosis and treatment. METHODS: We reviewed the clinical data of 6 patients with of external iliac artery vascular complications after renal transplantation from more than 2000 renal transplantation patients in the Transplantation Center of the Third Xiangya Hospital of Central South University from 2001 to 2013, and analyzed the clinical characteristics, diagnosis and treatment. RESULTS: The renal allograft was removed in 5 of the 6 patients due to repeated external iliac artery hemorrhage: 2 patients were replaced the external iliac artery with reversed autogenous great saphenous vein, 2 patients underwent the bilateral femoral artery bypass surgery, and 1 was repaired the external iliac artery directly. The other 1 was resected the renal allograft and the involved external iliac arteries due to fungal mass in the external iliac artery. Among the 6 patients, except 1 patient died after the surgery of the repair of the external iliac artery, the other 5 are all alive. CONCLUSION: Vascular replacement and artery bypass are effective methods for patients with external iliac artery vascular complications after kidney transplantation.

[Gene polymorphisms of CYP3A5 and MDR-1 in Hans renal transplant recipients in Hunan Province].

OBJECTIVE: To identify the polymorphisms of cytochrome P450 3A5 gene (CYP3A5) and multidrug resistance gene 1 (MDR-1) and their distributions in Hans renal transplant recipients in Hunan province, we analyzed the difference of the gene polymorphisms and distributions between Hunan province and 11 other provinces of China. METHODS: We collected 598 Hans renal transplant recipients who had operation or follow-up examination in 3rd Xiangya Hospital from Hunan province. We examined the gene polymorphisms of CYP3A5 and MDR-1 and compared their distributions with the data from 11 other provinces of China by chi-square test. RESULTS: There were CYP3A5*1/*1 genotype in 58 cases (9.7%), CYP3A5*1/*3 genotype in 251 cases (42.0%), CYP3A5*3/*3 genotype in 289 cases (48.3%); MDR-1 3435CC genotype in 238 cases (39.8%), MDR-1 3435CT genotype in 263 cases (44.0%), MDR-1 3435TT genotype in 97 cases (16.2%). Frequency of CYP3A5*1/*1 and *1/*3 genotypes of Hunan province was higher than the that from the 11 other provinces of China and the frequency of mutator *3 was lower. Frequency of MDR-1 3435CC and 3435CT genotypes of Hunan province was higher and the frequency of mutator T was lower than that from the 11 other provinces of China. CONCLUSIONS: There were significant difference in gene polymorphisms and distributions of CYP3A5 and MDR-1 between Hunan province and the 11 other provinces of China. It may be a guideline for us to use calcineurin inhibitor drugs in the early stage after renal transplantation.

Hepatic veins anatomy and piggy-back liver transplantation.

BACKGROUND: The piggy-back caval anastomosis technique is widely used in orthotopic liver transplantation although it carries an increased risk of complications, including outflow obstruction and Budd-Chiari syndrome. The aim of this study is to clarify the anatomy and variations of hepatic veins (HVs) draining into the inferior vena cava (IVC), and to classify the surgical techniques of piggy-back liver transplantation (PBLT) based on the anatomy of HVs which can reduce the occurrence of complications. METHODS: PBLT was performed in 248 consecutive cases at our hospital from January 2004 to August 2011. The anatomy of recipients' HVs was determined when removing the native diseased livers. Both anatomy of HVs and short HVs draining into the IVC were recorded. These data were collected and analyzed. RESULTS: We classified anatomic variations of HVs in the 248 livers into five types according to the way of drainage into the IVC: type I (trunk type of left and middle HVs), 142 (57.3%) patients; type II (trunk type of right and middle HVs), 54 (21.8%); type III (trunk type of left, middle and right HVs), 14 (5.6%); type IV (non-trunk type of left, middle and right HVs), of which, type IVa, 16 (6.5%), in the same horizontal plane; type IVb, 18 (7.3%), in different horizontal planes; and type V (segment type), 4 (1.6%). The patients whose HVs anatomy belonged to types I, II and III underwent classical piggy-back liver transplantation. Type IVa patients had classical PBLT via HV venoplasty prior to piggy-back anastomosis, while type IVb patients and type V patients could only have modified PBLT. CONCLUSION: This study demonstrates that HVs can be classified according to the anatomy of their drainage into the IVC and we can use this classification to choose the best operative approach to PBLT.

[Delayed graft function after DCD kidney transplantation: risk factors for and impact on transplantation].

OBJECTIVE: To evaluate the risk factors of delayed graft function (DGF) and its impact on renal transplantation from donation after cardiac death (DCD). METHODS: We conducted a retrospective study consisting of 48 subjects who underwent a DCD kidney transplantation from February 2010 to March 2012. We classified the recipients into two groups: an IGF (immediate graft function) group (n=30) and a DGF group (n=18), and analyzed the risk factors of DGF and its impact on transplantation. RESULTS: DGF occurred in 18 of the 48 (37.5%) kidneys from DCD donors, and the occurrence of DGF did not adversely influence the survival of patients (P=0.098) and graft (P=0.447). In the univariate analysis, the preoperative dialysis time of recipients (P<0.001), HLA mismatch site (P<0.001), the cause of brain death (P=0.011), BMI (P<0.001), preoperative serum creatinine of donors (P=0.0001), norepinephrine used in donors (P<0.001), warm ischema time (WIT) (P<0.001), cold ischema time (CIT) (P<0.001) showed significant differences. In the multivariate analysis, cerebral hemorrhage as the cause of brain death (P=0.022, OR=39.652), preoperative serum creatinine of donors≥177 µmol/L (P=0.008, OR=57.148) and the preoperative dialysis time of recipients≥12 months (P=0.060, OR=15.060) were independent risk factors for DGF development. CONCLUSION: The independent risk factors for DGF are the cause of brain death, the terminal creatinine level, and the preoperative dialysis time.

[Clinical analysis of 48 cases of kidney transplantation from cardiac death donors].

OBJECTIVE: To evaluate the recovery of patients with end-stage renal disease (ESRD) receiving kidney transplant from cardiac death donors, and to assess graft survival in China from this type of donor. METHODS: A total of 48 cases of patients with ESRD have received the kidneys from cardiac death donors in our hospital between February 2010 and March 2012. We retrospectively analyzed data on the preoperative and postoperative serum creatinine concentrations, on the survival of recipients and allografts with a view to investigating prognoses after this type of kidney transplant. RESULTS: Primary non-function (PNF) did not occur in any of the 48 recipients. Delayed graft function (DGF) occurred in 18 of 48 (37.5%) of kidneys from cardiac death donors, but the occurrence of DGF did not adversely influence patient's survival (P=0.098) or graft survival (P=0.447). Seven of 48 (14.6%) recipients lost their graft. Over a median follow-up period of 8 months (range 0.5-23 months), 39 of 41(95.1%) recipients' graft function had fully recovered. The actuarial graft and patient's survival rates at 1, 3, 6 and 12 months after transplantation were 95.7%, 93.0%, 90.0%, 87.5%, and 100%, 94.9%, 90%, 87.5%, respectively. CONCLUSION: As the legislation of donation after brain death (DBD) has not been ratified in China, the use of kidneys from cardiac death donors might be an effective way to increase the number of kidneys available for transplantation here. Our experience indicates good short- and mid-term outcomes with transplants from cardiac death donors.

Marijuana smoking and asthma: a protocol for a meta-analysis.

INTRODUCTION: Recent studies have raised the concern on the risk of asthma in marijuana smokers; however, the results remain controversial and warrant further investigation. With a growing number of marijuana smokers, examining the association between marijuana smoking and asthma and quantifying such association through meta-analysis have important implications for public health and clinical decision-making. In view of this, the present protocol aims to detail a comprehensive plan of meta-analysis on the association aforementioned. The findings are expected to strengthen the current knowledge base pertaining to the potential adverse effects of marijuana smoking on pulmonary health and to facilitate the development of prevention strategies for asthma. METHODS AND ANALYSIS: The MEDLINE/PubMed, Web of Science and EMBASE databases will be searched systematically from inception to 1 September 2021 to retrieve the relevant observational studies focusing on the association between marijuana smoking and asthma. Both unadjusted and adjusted effect sizes, such as OR, relative risk, HR and the corresponding 95% CIs will be extracted for pooled analyses. Heterogeneity and publication bias across the included studies will be examined. The Newcastle-Ottawa Quality Scale will be used to assess the quality and risk of bias. Statistical software Review Manager V.5.3 and Stata V.11.0 will be used for statistical analyses. ETHICS AND DISSEMINATION: Since no private and confidential patient data will be included in the reporting, approval from an ethics committee is not required. The results will be published in a peer-reviewed journal or disseminated in the relevant conferences. The study raises no ethical issue. OSF REGISTRATION NUMBER: 10.17605/OSF.IO/UPTXC.

Posttransplant-Alloantibodies Against MICA Antigens Associated With Decreased Long-Term Allograft Survival of Kidney Transplant Recipients.

BACKGROUND: Previous evidence showed that antibodies against major histocompatibility complex class I-related chain A (MICA) could lead to antibody-mediated rejection in kidney transplantation in case where the patients had no alloantibodies against HLA. However, the effects of posttransplant anti-MICA antibodies on long-term renal allograft survival and function remained unsettled. We tested the posttransplant anti-MICA antibodies in 150 kidney transplant patients. The aim of this study was to compare the long-term graft survival and function between patients who were MICA positive and those who were negative. METHODS: The posttransplant serum samples from 150 patients receiving kidney transplantation in our center from 2012 to 2013 were tested for MICA antibodies and HLA antibodies by Luminex single antigen array technology. Graft survival and function were followed up for a mean time of 74.2 months. The research was conducted in accordance with the Helsinki Congress and the Declaration of Istanbul. RESULTS: Of the 150 patients, 38 (25.3%) were sensitized against MICA after transplantation. The anti-MICA antibodies-positive (anti-MICA+) group had a worse long-term renal allograft survival than that of anti-MICA-negative (anti-MICA-) group (P = .029), even when stratified by posttransplant HLA sensitization status or donor source. Anti-MICA antibodies also had a detrimental impact on renal allograft function, but only at 1 year posttransplantation (estimated glomerular filtration rates at 1 year: anti-MICA+ 66.6 mL/min/1.73 m2 vs anti-MICA- 78.7 mL/min/1.73 m2; P = .023). CONCLUSION: Posttransplant anti-MICA antibodies were associated with decreased long-term renal allograft survival and short-term renal allograft function.

LED Light Quality of Continuous Light before Harvest Affects Growth and AsA Metabolism of Hydroponic Lettuce Grown under Increasing Doses of Nitrogen.

To study the effects of light quality of continuous light before harvest on the growth and ascorbic acid (AsA) metabolism of lettuce (Lactuca sativa L.) grown under relative high nitrogen level, lettuce plants grown under different nitrogen levels (8, 10 and 12 mmol·L-1) were subjected to continuous light with different red: blue light ratios (2R:1B and 4R:1B) before harvest. The results showed that the shoot fresh weight of lettuce under 12 mmol·L-1 nitrogen level was significantly higher than that under other treatments. There were no significant differences in shoot dry weight, root fresh weight, root dry weight, soluble sugar content, nitrate content and AsA content in leaves among the treatments at different nitrogen levels. The content of AsA in leaves was significantly higher than that in petioles before and after continuous light. Under the same nitrogen level, the fresh weight of lettuce under continuous light quality 4R:1B was significantly higher than that under other treatments. The content of AsA in lettuce leaves increased in different degrees after continuous light before harvest. High yield and AsA content could be obtained by 72 h continuous light with red and blue light 4R:1B at 12 mmol·L-1 nitrogen level. After continuous light, the content of AsA increased significantly due to the increase of the ratio of red light and nitrogen level, which increased the activities of L-galactono-1,4-lactone dehydrogenase (GalLDH) and dehydroascorbic acid reductase (DHAR) involved in AsA synthesis and in the recycling of DHAR to AsA respectively.

Light Quality Affected the Growth and Root Organic Carbon and Autotoxin Secretions of Hydroponic Lettuce.

Light is a crucial environmental signal and photosynthetic energy for plant growth, development, and primary and secondary metabolism. To explore the effects of light quality on the growth and root exudates of hydroponic lettuce (Lactuca sativa L.), white LED (W, control) and four the mixtures of red (R) and blue (B) LED with different R/B light intensity ratios (R/B = 2, 2R1B; R/B = 3, 3R1B; R/B = 4, 4R1B; and R/B = 8, 8R1B) were designed. The results showed that the biomass of lettuce under 8R1B and W treatments was higher than that under other light quality treatments. The photosynthetic rate (Pn) under red and blue light was significantly higher than that of white light. Total root length, root surface area, and root volume were the highest under 8R1B. 4R1B treatment significant increased root activity by 68.6% compared with W. In addition, total organic carbon (TOC) content, TOC content/shoot dry weight, TOC content/root dry weight, and TOC content/root surface area were the highest under 4R1B. Moreover, 8R1B treatment reduced the concentration of benzoic acid and salicylic acid, and the secretion ability of benzoic acid and salicylic acid by per unit root surface area and accumulation by per unit shoot dry weight. In addition, 2R1B and 3R1B reduced the secretion ability of gallic acid and tannic acid by per unit root surface area and accumulation by per unit shoot dry weight. In conclusion, this study showed that the secretion of autotoxins could be reduced through the mediation of red and blue light composition of LEDs in a plant factory. In terms of autotoxin secretion reduction efficiency and yield performance of lettuce, 8R1B light regime is recommended for practical use.

Regulation of Ascorbate Accumulation and Metabolism in Lettuce by the Red:Blue Ratio of Continuous Light Using LEDs.

Ascorbate (AsA), an antioxidant that cannot be synthesized and stored by the human body, plays an essential role in the proper functioning of both plants and humans. With the goal of increasing the AsA level in lettuce, the effects of different ratios of red (R) to blue (B) light (75R:25B, 50R:50B, and 25R:75B) on AsA pool sizes as well as the transcript levels and activities of key enzymes involved in AsA metabolism were constantly monitored for 12 days under continuous light (200 µmol⋅m-2⋅s-1) from LEDs. The results showed that lettuce biomass was positively correlated with the ratio of red light, while the AsA pool size had a positive correlation with the ratio of blue light during the whole experiment. The 25R:75B treatment increased the expression of genes involved in AsA biosynthesis (GMP, GME, GGP, GPP, GLDH) and regeneration (APX, MDHAR, DHAR, and GR) on day 3 but only significantly elevated the activities of enzymes involved in AsA regeneration (APX, MDHAR, DHAR, and GR) subsequently. AsA regeneration enzymes (MDHAR, DHAR and GR) had greater correlations with the AsA level than the AsA synthesis enzyme (GLDH). Thus, it is concluded that a high ratio of blue light elevated the AsA level mainly by promoting AsA regeneration rather than biosynthesis. Taken together, altering the red:blue ratio of continuous light from high to low before harvest is recommended for lettuce cultivation to achieve both high yield and high quality.

The botanical origin and antioxidant, anti-BACE1 and antiproliferative properties of bee pollen from different regions of South Korea.

BACKGROUND: Bee pollen (BP) has been used as a traditional medicine and food diet additive due to its nutritional and biological properties. The potential biological properties of bee pollen vary greatly with the botanical and geographical origin of the pollen grains. This study was conducted to characterize the botanical origin and assess the antioxidant effects of ethanol extracts of 18 different bee pollen (EBP) samples from 16 locations in South Korea and their inhibitory activities on human ß-amyloid precursor cleavage enzyme (BACE1), acetylcholinesterase (AChE), human intestinal bacteria, and 5 cancer cell lines. METHODS: The botanical origin and classification of each BP sample was evaluated using palynological analysis by observing microscope slides. We measured the biological properties, including antioxidant capacity, inhibitory activities against human BACE1, and AChE, and antiproliferative activities toward five cancer cell lines, of the 18 EBPs. In addition, the growth inhibitory activities on four harmful intestinal bacteria, six lactic acid-producing bacteria, two nonpathogenic bacteria, and an acidulating bacterium were also assessed. RESULTS: Four samples (BP3, BP4, BP13 and BP15) were found to be monofloral and presented four dominant pollen types: Quercus palustris, Actinidia arguta, Robinia pseudoacacia, and Amygdalus persica. One sample (BP12) was found to be bifloral, and the remaining samples were considered to be heterofloral. Sixteen samples showed potent antioxidant activities with EC50 from 292.0 to 673.9 µg mL- 1. Fourteen samples presented potent inhibitory activity against human BACE1 with EC50 from 236.0 to 881.1 µg mL- 1. All samples showed antiproliferative activity toward the cancer cell lines PC-3, MCF-7, A549, NCI-H727 and AGS with IC50 from 2.7 to 14.4 mg mL- 1, 0.9 to 12.7 mg mL- 1, 5.0 to > 25 mg mL- 1, 2.7 to 17.7 mg mL- 1, and 2.4 to 8.7 mg mL- 1, respectively. In addition, total phenol and flavonoid contents had no direct correlation with antioxidant, anti-human BACE1, or antiproliferative activities. CONCLUSION: Fundamentally, Korean bee pollen-derived preparations could be considered a nutritional addition to food to prevent various diseases related to free radicals, neurodegenerative problems, and cancers. The botanical and geographical origins of pollen grains could help to establish quality control standards for bee pollen consumption and industrial production.

Non-Mitogenic Fibroblast Growth Factor 1 Enhanced Angiogenesis Following Ischemic Stroke by Regulating the Sphingosine-1-Phosphate 1 Pathway.

Ischemic strokes account for about 80% of all strokes and are associated with a high risk of mortality. Angiogenesis of brain microvascular endothelial cells may contribute to functional restoration following ischemia. Fibroblast growth factor 1 (FGF1), a member of FGF superfamily, involved in embryonic development, angiogenesis, wound healing, and neuron survival. However, the mitogenic activity of FGF1 is known to contribute to several human pathologies, thereby questioning the safety of its clinical applications. Here, we explored the effects and mechanism of action of non-mitogenic FGF1 (nmFGF1) on angiogenesis in mice after ischemia stroke and an oxygen-glucose deprivation (OGD)-induced human brain microvascular endothelial cells (HBMECs) injury model. We found that intranasal administration nmFGF1 significantly promoted angiogenesis in mice after stroke, and significantly increased the formation of matrigel tube and promoted scratch migration in a dose-dependent manner in OGD-induced HBMECs in vitro. However, the co-administration of an FGF receptor 1 (FGFR1)-specific inhibitor PD173074 significantly reversed the effects of nmFGF1 in vitro, suggesting that nmFGF1 functions via FGFR1 activation. Moreover, nmFGF1 activated sphingosine-1-phosphate receptor 1 (S1PR1, S1P1) in mice after stroke in vivo. S1P1 protein antagonist VPC23019 and agonist FTY720 were used to confirm that nmFGF1 promotes angiogenesis in vitro partially through the S1P1 pathway. OGD induced downregulation of S1P1 expression. The S1P1 antagonist VPC23019 blocked the stimulatory effects of nmFGF1, whereas the S1P1 agonist FTY720 exerted effects comparable with those of nmFGF1. Furthermore, PD173074 reversed the effect of nmFGF1 on upregulating S1P1 signaling. In conclusion, nmFGF1 enhanced angiogenesis in mice following stroke and OGD-induced HBMECs through S1P1 pathway regulation mediated via FGFR1 activation. This new discovery suggests the potential therapeutic role of nmFGF1 for the treatment of ischemic strokes.

Morphological and Physiological Stress Responses of Lettuce to Different Intensities of Continuous Light.

In this study, specific dynamic changes in growth, oxidative stress, ascorbate metabolism, and chlorophyll fluorescence were monitored during 12 days in lettuce plants exposed to continuous light (CL) of different intensities: low light (LL, 100 µmol·m-2·s-1), medium light (ML, 200 µmol·m-2·s-1), and high light (HL, 300 µmol·m-2·s-1). Lettuce plants grown under CL of higher light intensity gained greater biomass, dry weight ratio, root/shoot ratio, and specific leaf FW, but not leaf area. Both the reactive oxygen species (ROS) production and the lipid peroxidation degree, measured in terms of the malondialdehyde (MDA) levels, were progressively enhanced by increasing the light intensity of CL. Overall, the pool sizes of ascorbate (AsA) and glutathione, as well as the activities of enzymes involved in AsA metabolism, had positive correlations with light intensity under CL. Ascorbate peroxidase and dehydroascorbate reductase presented the maximal and minimal responses to light intensity, respectively, among all the studied enzymes. After 6 days under CL, ML and HL intensity caused reversible photoinhibition, represented by lower values of maximum quantum efficiency (F v /F m), effective quantum yield (ΦPSII), and photochemical quenching (qP) and a higher value of non-photochemical quenching (qN). However, this photoinhibition recovered on day 12 with increasing of F v /F m, ΦPSII, and qP. Taken together, under ML and HL conditions, greater AsA level could help maintain photosynthetic efficiency by elevating excess excitation energy dissipation, though ROS accumulation and lipid peroxidation could not be prevented in the long-term. Likewise, there was no dark period under LL condition, but no photooxidative stress was observed in lettuce. Thus, it is concluded that photooxidative stress induced by CL can be attributed to excessive daily light integral instead of circadian asynchrony.

Potential Prognostic and Diagnostic Values of CDC6, CDC45, ORC6 and SNHG7 in Colorectal Cancer.

BACKGROUND: Colorectal cancer (CRC) is a common human malignancy. The aims of this study are to investigate the gene expression profile of CRC and to explore potential strategy for CRC diagnosis, therapy and prognosis. METHODS: We use affy and Limma package of Bioconductor R to do differential expression genes (DEGs) and differential expression lncRNAs (DELs) analysis from the gene datasets (GSE8671, GSE21510, GSE32323, GSE39582 and TCGA) respectively. Then, DEGs were analyzed by GO and KEGG pathway and Kaplan-Meier survival curve and Cox regression analyses were used to find aberrantly expressed genes associated with survival outcome of CRC patients. Real-time PCR assay was used to verify the aberrantly expressed genes expression in CRC samples. RESULTS: 306 up-regulation and 213 down-regulation common DEGs were found. A total of 485 DELs were identified, of which 241 up-regulated and 244 down-regulated. Then, GO and KEGG pathway analyses showed that DEGs were involved in cell cycle, mineral absorption, DNA replication, and Nitrogen metabolism. Among them, Kaplan-Meier survival curve and Cox regression analyses revealed that CDC6, CDC45, ORC6 and SNHG7 levels were significantly associated with survival outcome of CRC patients. Finally, real-time PCR assay was used to verify that the CDC6, CDC45, ORC6 and SNHG7 expression were up-regulated in 198 CRC samples compared with the expression levels in individual-matched adjacent mucosa samples. CONCLUSION: CDC6, CDC45, ORC6 and SNHG7 are implicated in CRC initiation and progression and could be explored as potential diagnosis, therapy and prognosis targets for CRC.

Poly(amine-co-ester) nanoparticles for effective Nogo-B knockdown in the liver.

Degradable poly(amine-co-ester) (PACE) terpolymers hold tremendous promise for siRNA delivery because these materials can be formulated into delivery vehicles with highly efficient siRNA encapsulation, providing effective knockdown with low toxicity. Here, we demonstrate that PACE nanoparticles (NPs) provide substantial protein knockdown in human embryonic kidney cells (HEK293) and hard-to-transfect primary human umbilical vein endothelial cells (HUVECs). After intravenous administration, NPs of solid PACE (sPACE)-synthesized with high monomer content of a hydrophobic lactone-accumulated in the liver and, to a lesser extent, in other tissues. Within the liver, a substantial fraction of sPACE NPs were phagocytosed by liver macrophages, while a smaller fraction of NPs accumulated in hepatic stellate cells and liver sinusoidal endothelial cells, suggesting that sPACE NPs could deliver siRNA to diverse cell populations within the liver. To test this hypothesis, we loaded sPACE NPs with siRNA designed to knockdown Nogo-B, a protein that has been implicated in the progression of alcoholic liver disease and liver fibrosis. These sPACE:siRNA NPs produced up to 60% Nogo-B protein suppression in the liver after systemic administration. We demonstrate that sPACE NPs can effectively deliver siRNA therapeutics to the liver to mediate protein knockdown in vivo.