Transcriptomic Profile Identifies Hippocampal Sgk1 as the Key Mediator of Ovarian Estrogenic Regulation on Spatial Learning and Memory and Aß Accumulation.

Previous studies have shown that ovarian estrogens are involved in the occurrence and pathology of Alzheimer's disease (AD) through regulation on hippocampal synaptic plasticity and spatial memory; however, the underlying mechanisms have not yet been elucidated at the genomic scale. In this study, we established the postmenopausal estrogen-deficient model by ovariectomy (OVX). Then, we used high-throughput Affymetrix Clariom transcriptomics and found 143 differentially expressed genes in the hippocampus of OVX mice with the absolute fold change ≥ 1.5 and P < 0.05. GO analysis showed that the highest enrichment was seen in long-term memory. Combined with the response to steroid hormone enrichment and GeneMANIA network prediction, the serum and glucocorticoid-regulated kinase 1 gene (Sgk1) was found to be the most potent candidate for ovarian estrogenic regulation. Sgk1 overexpression viral vectors (oSgk1) were then constructed and injected into the hippocampus of OVX mice. Morris water maze test revealed that the impaired spatial learning and memory induced by OVX was rescued by Sgk1 overexpression. Additionally, the altered expression of synaptic proteins and actin remodeling proteins and changes in CA1 spine density and synapse density induced by OVX were also significantly reversed by oSgk1. Moreover, the OVX-induced increase in Aß-producing BACE1 and Aß and the decrease in insulin degrading enzyme were significantly reversed by oSgk1. The above results show that multiple pathways and genes are involved in ovarian estrogenic regulation of the function of the hippocampus, among which Sgk1 may be a novel potent target against estrogen-sensitive hippocampal dysfunctions, such as Aß-initiated AD.

LH upregulates connexin 43 expression in granulosa cells by activating the Wnt/ß-catenin signalling pathway.

Connexin (Cx) 43 is the most widely expressed gap junction protein in follicle granulosa cells and plays an important role in follicle development and growth. The aims of this study were to investigate the effects of LH on the expression of Cx43 and key proteins in the downstream Wnt-ß/catenin signalling pathway and to explore the mechanism underlying the regulation of Cx43 expression in granulosa cells. Primary culture granulosa cells were obtained from 21-day-old Sprague-Dawley rats, and were treated with different concentrations of LH (150, 300 and 600 IU L-1). Cx43 expression in granulosa cells was detected using immunofluorescence. Western blotting was used to detect the expression of Cx43, ß-catenin and Axin2 proteins (Axin2 is a protein that in humans is encoded by the AXIN2 gene, which presumably plays an important role in the regulation of the stability of ß-catenin in the Wnt signaling pathway) in granulosa cells with and without FH535 treatment (a Wnt/ß-catenin signalling pathway inhibitor). Cx43 expression was detected in the cytoplasm and cell membrane of granulosa cells. Treatment with a high concentration of LH (300 IU L-1) increased the expression of ß-catenin and Axin2, as well as that of Cx43. FH535 treatment reduced the LH-induced increases in Cx43, ß-catenin and Axin2. These results indicate that LH upregulates Cx43 expression in granular cells by activating the Wnt/ß-catenin signalling pathway.

Exogenous luteinizing hormone promotes ovarian survival and function during cryopreservation and transplantation.

Cryopreservation and transplantation of the ovarian tissue is an alternative method by which malignant tumor survivors can recover fertility. Previously, it was reported that follicle stimulating hormone (FSH) promoted the survival and functioning of the ovarian tissue after in vitro cultivation. In this study, the expression of the luteinizing hormone receptor (LHR) was observed on the granule cell membrane after luteinizing hormone (LH) (0.3 IU/mL) was supplied as an exogenous hormone into the cultivation medium during ovarian vitrification in the postnatal period (PND) (1, 7, 14, 21, 28, 42, and 56 days PND). The expression of vascular endothelial growth factor (VEGF) and Connexins (Cx), and the recovery of ovarian functions were then assessed in mice models. The results showed that LH increased the production of normal follicles, and upregulated the expression of VEGF, Cx37, and Cx43 in vitrified ovaries. LH administration also shortened the recovery time of the estrus cycle in mice models. Additionally, no difference was observed in the rate of pregnancy and size of the first litter between the experimental and control groups. In conclusion, LH could promote the survival and functioning of the ovaries by upregulating the expression of VEGF, Cx43, and Cx37 during ovarian cryopreservation and transplantation.

[Role of mimecan in development of atherosclerosis induced by increased blood pressure variability].

OBJECTIVE: To examine the changes of mimecan protein expression in development of atherosclerosis induced by sinoaortic denervation, and to explore the effects of mimecan knock down on the proliferation and migration of vascular smooth muscle cells.
 Methods: The animals were randomly divided into a sham group and a model group (n=8 in each group). The rat model of blood pressure variability was established by sinoaortic denervation, and the hemodynamic indexes were recorded 20 weeks after the surgery to confirm the success of the model. The thoracic aorta was excised and stained with immunohistochemistry to observe the pathological changes of smooth muscle tissues and the changes of mimecan expression. The mice vascular smooth muscle cells were isolated, and which were treated with mimecan siRNA to knock down the mimecan expression. The cell proliferation was observed by 5-ethynyl-2'-deoxyuridine (Edu) in corporation test and the changes of cell migration was observed by wound healing test.
 Results: Twenty weeks after sinoaortic denervation, the blood pressure variability in the model group was significantly increased compared with that in the sham group, suggesting the model was successfully established. In addition, the increased blood pressure variability in the model group promoted the proliferation and migration of the vascular smooth muscle cells in thoracic aorta, while the expression of mimecan protein was significantly decreased. In in vitro assays, the knock down of mimecan in mice vascular smooth muscle cells could promote the cell proliferation and migration.
 Conclusion: Mimecan plays a protective role in the development of sinoaortic denervation induced atherosclerosis through amechanism involving suppression of the proliferation and migration of vascular smooth muscle cells.

Antinociceptive effects of gentiopicroside on neuropathic pain induced by chronic constriction injury in mice: a behavioral and electrophysiological study.

Gentiopicroside (Gent) is promising as an important protective secoiridoid compound against pain. The present study was designed to investigate whether administration of Gent would alleviate the expression of nociceptive behaviors and whether it would cause the relevant electrophysiological changes in a chronic constriction injury (CCI) model of neuropathic pain in mice. Gent was administered from the seventh day after surgery for 8 consecutive days. Behavioral parameters and sciatic functional index were assessed immediately before surgery and on days 7, 8, 10, 12, and 14 post-CCI, and electrophysiological activities of sciatic nerve were recorded immediately after the behavioral test on the last day. The present study has shown that administration of Gent (at a dose of 50 and 100 mg/kg) increased behavioral parameters from day 8 compared with the CCI-NS group. Electrophysiological data indicated that CCI caused a significant reduction in nerve conduction velocities in the sciatic nerves and the amplitudes of compound action potential, while Gent at a dose of 50 or 100 mg/kg caused a significant recovery of electrophysiological changes induced by CCI. Our data indicated that Gent has antinociceptive effects on neuropathic pain induced by CCI.

Oxysophocarpine Ameliorates Carrageenan-induced Inflammatory Pain via Inhibiting Expressions of Prostaglandin E2 and Cytokines in Mice.

Oxysophocarpine is an alkaloid extracted from Sophora alopecuroides. We investigated the analgesic effect of oxysophocarpine on carrageenan-induced inflammatory pain in mice, in order to explore its possible mechanisms. Mouse ear swelling tests and carrageenan-induced paw edema tests were used to investigate the effects of oxysophocarpine on inflammatory pain in mice. Morphological changes on inflamed paw sections were measured by hematoxylin-eosin staining. The mRNA and protein expression of extracellular signal-regulated kinase, phosphorylation of extracellular signal-regulated kinase 1/2, cyclooxygenase-2, tumor necrosis factor α, interleukin-1 beta, interleukin-6 and prostaglandin E2 were investigated by real-time quantitative polymerase chain reaction, immunohistochemistry, western-blot and enzyme-linked immunosorbent assay. In our results, oxysophocarpine shows a significant anti-inflammatory effect in the mouse ear swelling test. Oxysophocarpine also significantly reduced the paw edema volume and improved mechanical allodynia threshold value on carrageenan-induced inflammatory pain, as well as relieved paw tissues inflammatory damage and reduced the numbers of neutrophils in mice. Oxysophocarpine significantly suppressed over-expression of cyclooxygenase-2, tumor necrosis factor α, interleukin-1 beta, interleukin-6 and prostaglandin E2, and inhibited the over-phosphorylation of extracellular signal-regulated kinase 1/2. Based on these findings we propose that oxysophocarpine attenuates inflammatory pain by suppressing the levels of phosphorylation of extracellular signal-regulated kinase 1/2, cyclooxygenase-2, prostaglandin E2, tumor necrosis factor α, interleukin-1 beta and interleukin-6.

Aloperine attenuated neuropathic pain induced by chronic constriction injury via anti-oxidation activity and suppression of the nuclear factor kappa B pathway.

OBJECTIVE: To investigate whether aloperine (ALO) has antinociceptive effects on neuropathic pain induced by chronic constriction injury, whether ALO reduces ROS against neuropathic pain, and what are the mechanisms involved in ALO attenuated neuropathic pain. METHODS: Mechanical and cold allodynia, thermal and mechanical hyperalgesia and spinal thermal hyperalgesia were estimated by behavior methods such as Von Frey filaments, cold-plate, radiant heat, paw pressure and tail immersion on one day before surgery and days 7, 8, 10, 12 and 14 after surgery, respectively. In addition, T-AOC, GSH-PX, T-AOC and MDA in the spinal cord (L4/5) were measured to evaluate anti-oxidation activity of ALO on neuropathic pain. Expressions of NF-κB and pro-inflammatory cytokines (TNF-α, IL-6, IL-1ß) in the spinal cord (L4/5) were analyzed by using Western blot. RESULTS: Administration of ALO (80mg/kg and 40mg/kg, i.p.) significantly increased paw withdrawal threshold, paw pressure, paw withdrawal latencies, tail-curling latencies, T-AOC, GSH-PX and T-SOD concentration, reduced the numbers of paw lifts and MDA concentration compared to CCI group. ALO attenuated CCI induced up-regulation of expressions of NF-κB, TNF-α, IL-6, IL-1ß at the dose of 80mg/kg (i.p.). Pregabalin produced similar effects serving as positive control at the dose of 10mg/kg (i.p.). CONCLUSION: ALO has antinociceptive effects on neuropathic pain induced by CCI. The antinociceptive effects of ALO against neuropathic pain is related to reduction of ROS, via suppression of NF-κB pathway.

Relaxative effect of core licorice aqueous extract on mouse isolated uterine horns.

CONTEXT: Primary dysmenorrhea is one of the most frequent gynecological disorders in young women. Chinese herbal medicine has the advantage in terms of multi-targeting efficacy, lower toxicity, as well as lower cost. Core licorice is the hard and atropurpureus heart part in root and rootstock of Glycyrrhiza uralensis Fisch (Leguminosae), having a therapeutic effect on dysmenorrhea. OBJECTIVE: This experiment indicated the spasmolytic effect of core licorice aqueous extract (CLE) on spontaneous rhythmic contractions and spasmogen-provoked contractions of stilbestrol primed, estrogen-dominated, non-pregnant mouse isolated uterine horns and its spasmolytic mechanism. MATERIALS AND METHODS: We investigated the spasmolytic effect of CLE (0.025-0.1 mg/mL) on spontaneous contractions and potassium chloride (KCl, 40 mM), acetylcholine (ACh, 5 µg/mL), carbachol (CCh, 5 µg/mL), oxytocin (OT, 2 U/L) or bradykinin (5 ng/mL)-provoked contractions of mouse isolated uterine horns. Contractions were recorded by tension force transducers using Biolap 420F software on a PC. RESULTS: Our present study showed that graded, escalated concentrations of CLE (0.025-0.1 mg/mL) significantly inhibited the amplitude of spontaneous phasic contractions (15.03-55.10%), as well as the contractions produced by KCl (40 mM; 20.16-53.99%), ACh (5 µg/mL; 14.65-48.32%), CCh (5 µg/mL; 38.40-76.70%), OT (2 U/L; 21.53-58.49%) or bradykinin (5 ng/mL; 58.01-79.44%) of the estrogen-dominated isolated mice uterine horn preparations in a concentration-related manner. DISCUSSION AND CONCLUSION: The spasmolytic effect of CLE observed in the present study lends pharmacological support to the traditional use of core licorice in the management, control and treatment of primary dysmenorrhea.

Ovarian toxicity of e-cigarette liquids: Effects of components and high and low nicotine concentration e-cigarette liquid in vitro.

INTRODUCTION: Electronic cigarette use has become increasingly popular, with potential consequences for reproductive health. We aimed to investigate the effects of different components of e-liquid on the ovary and compare the impact of low nicotine concentration e-liquids (LN e-liquids) and high nicotine concentration e-liquids (HN e-liquids) on ovarian toxicity. METHODS: A total of 378 rat ovaries were divided into seven groups, including control (no intervention), nicotine (0.05 mg/mL), flavoring (0.25 µL/mL), propylene glycol (PG) (2.5 µL/mL), vegetable glycerin (VG) (2.0 µL/mL), LN e-liquid (0.05 mg nicotine + 0.25 µL flavoring + 2.5 µL PG + 2.0 µL VG + 0.25 µL distilled water/mL medium) and HN e-liquid groups (0.05 mg nicotine + 0.05 µL flavoring + 0.5 µL PG + 0.4 µL VG + 0.05 µL distilled water/mL medium). After three hours of in vitro culture, ovarian morphology, oxidation levels [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA)], and apoptosis levels [factor related apoptosis (Fas), Cyt-c, Caspase-9, Caspase-3] were analyzed. RESULTS: Our findings indicate that nicotine has limited impact on the ovary, while flavoring, PG, and VG all cause ovarian damage including morphological damage, disruption of oxidative balance and promotion of apoptosis, with VG having the most significant effect. Moreover, LN e-liquids may lead to more severe ovarian damage than HN e-liquids at an equal intake of total nicotine. CONCLUSIONS: Our study highlights that in e-liquid formula, nicotine has a limited effect on the ovaries, but flavoring, PG, and VG all cause damage to the ovaries, with VG the most damaging. At a consistent level of total nicotine intake, e-liquids with low nicotine concentrations cause more damage to the ovaries than those with high nicotine concentrations. These findings contribute to a better understanding of the impact of e-liquids on ovarian health and have important implications for public health policy.

Effects of HMG on revascularization and follicular survival in heterotopic autotransplants of mouse ovarian tissue.

Ovarian tissue transplantation is now considered as a procedure to preserve the fertility of young women patients undergoing cancer therapy. The present study investigated the effects and mechanism of human menopausal gonadotrophin (HMG) intervention on vascular remoulding in ovarian heterotopic autotransplantation. Ovaries of 8-week-old mice were cultured in vitro with different concentrations of HMG for 3h for measuring the expression of vascular endothelial growth factor (VEGF). The cultured ovaries were implanted under the kidney capsule and removed 24, 36, 48 h or 1 month after transplantation. Revascularization, fluid exudation and the number of surviving ovarian follicles were observed. The results showed that VEGF was increased 1.6-6.5 times in the HMG intervention groups. Revascularization appeared 24-36 h after transplantation and was earlier than that of the control. Fluid exudation increased incrementally with increasing HMG concentrations. The total number of surviving ovarian follicles was increased by 1.2-1.5 times in the HMG 0.15 IU/ml group as compared with the other groups 1 month after transplantation. It is concluded that intervention with HMG in vitro before transplantation could improve the blood supply reconstruction and survival of the autotransplanted ovarian follicles, which might be associated with increased VEGF expression.

Electrochemical sensor for the simultaneous detection of CA72-4 and CA19-9 tumor markers using dual recognition via glycosyl imprinting and lectin-specific binding for accurate diagnosis of gastric cancer.

The simultaneous identification and detection of multiple tumor markers provide more pathological information for the accurate diagnosis of cancer. In this study, a novel glycosyl imprinted sensor for the simultaneous detection of tumor markers CA19-9 and CA72-4 was prepared by combining the specific recognition of the glycosylated imprinting technique and lectin-characteristic glycan chains. The imprinted membrane was fabricated by electropolymerization using the characteristic glycopeptide STn on the surface of CA72-4 and the characteristic glycopeptide SLea on the surface of CA19-9 as template molecules and 2-aminophenylboronic acid as the functional monomer. To further improve the recognition efficiency, the specific binding of lectins to glycosyl chains was introduced. Sambucus nigra agglutinin I (SNA I) was labeled with cysteine, and Maackia amurensis lectin II (MAL II) was labeled with ferrocene. According to the specific binding of SNA to the Neu5Acα2-6GalNAcα-glycan chain in STn and MAL to α2, 3 sialic acid in SLea, CA72-4, and CA19-9 could be determined, respectively, after recombination between the glycosyl groups and GIP. The sensor shows high sensitivity to CA72-4 and CA19-9 in the concentration range of 0.005-200.0 U/mL, and it has been successfully applied to the detection of CA72-4 and CA19-9 in serum samples. The sensor provides a simple, fast, and low-cost alternative for accurate diagnosis of gastric cancer.

Effect of e-cigarette refill liquid on follicular development and estrogen secretion in rats.

INTRODUCTION: Electronic cigarettes (e-cigarettes) have recently become popular as an alternative to conventional cigarettes. The aim of this study is to investigate the effects of e-cigarette refill liquid (e-liquid) on follicular development and estrogen secretion in rats and whether it is related to the Hippo signaling pathway, a pathway that can regulate follicle growth. METHODS: Ovaries from 21- and 35-day-old rats were divided into three groups: control (no intervention), 0.05 mg, and 0.5 mg (e-liquids containing 0.5 mg and 5 mg of nicotine/kg). The rates were cultured for three hours in vitro. At the end of culture, HE staining was performed to observe the follicle morphology and calculate the percentage of normal follicles, and the expression of Yes-associated protein (YAP, target factors of the Hippo signaling pathway) and CYP19 (aromatase, a key enzyme in estrogen synthesis) were observed by immunohistochemistry. Western blotting was performed to detect the expression levels of CYP19, YAP, phosphorylated YAP (PYAP), large tumor suppressor 2 (LATS2, factors upstream of YAP in the Hippo signaling pathway), and phosphorylated LATS2 (PLATS2). Estrogen concentrations were determined using ELISA. RESULTS: HE staining showed that the percentage of normal follicles decreased, and immunohistochemistry showed that the expression of CYP19 and YAP significantly decreased after e-liquid intervention. ELISA showed that the estrogen concentration in the ovaries decreased after e-liquid intervention. Western blot results indicated that CYP19, LATS2, and YAP expression, decreased after e-liquid intervention, but PLATS2 and PYAP expression increased. CONCLUSIONS: We found that the e-liquids may impair the development of rat ovarian follicles and reduce estrogen secretion through Hippo signaling pathway.

[Effect of oxysophoridine on electric activities and its power spectrum of reticular formation in rats].

OBJECTIVE: To observe the effect of oxysophoridine (OSR) on the EEG and its power spectrum of reticulum formation in mesencephalon of anaesthetized rat. METHOD: Utilizing the technique of brain stereotactic apparatus, electrodes were implanted into reticulum formation of mesencephalon. Monopolar lead and computerized FFT technique were employed to record and analyse the index of EEG, power spectrum and frequency distribution in order to study the effect of oxysophoridine on the bioelectricity change of mesencephalon reticulum formation in rats. RESULT: After administrating(icy) with oxysophoridine at the dose of 2.5,5, 10 mg/rat, the EEG of mesencephalon reticulum formation mainly characterized with low amplitude and slow waves accompanied by spindle-formed sleeping waves with a significant decrease of total power of EEG (P < 0.05) while the ratio of theta, alpha waves increased in total frequency of rats (P < 0.05). CONCLUSION: Oxysophoridine possesses central inhibitory effects and its inhibitory mechanism may associate with the reduction of bioelectricity in mesencephalon reticulum formation. Mesencephalon reticulum formation may serve as one part of the structure serving as the circuit conducting the central inhibitory effect of oxysophoridine. [Key words] oxysophoridine; reticulum formation; electroencephalogram (EEG) ; rats

[Preparation of anti-GPC3 single chain antibody for targeted detection of hepatocellular carcinoma].

Glypican-3 (GPC3) is a key member of Glypican family and plays an important role in the development, angiogenesis and metastasis of hepatocellular carcinoma (HCC). Most HCC overexpresses GPC3, but GPC3 is hardly detected in normal adult liver and benign liver lesions, so it is regarded as a highly specific diagnostic marker and an ideal therapeutic target for HCC. In this study, we cloned the heavy and light chain variable region gene from the monoclonal antibody targeted to GPC3 screened in the previous stage, and linked it with a segment of flexible peptide (Linker) to obtain the single chain antibody against GPC3. The single chain antibody gene was cloned into vector for prokaryotic expression and purified to obtain high purity protein. Detection shows that the single-chain antibody produced by us has the same binding activity with the full-length antibody, and can accurately target the tumor site of Huh7 tumor-bearing model mice after coupling Cy5.5 fluorescence, suggesting that the single-chain antibody has the potential to realize multi-directional liver cancer precise surgical navigation under the guidance of a probe.

A strategy for construction of highly sensitive glycosyl imprinted electrochemical sensor based on sandwich-like multiple signal enhancement and determination of neural cell adhesion molecule.

A novel electrochemical sensor for a neural cell adhesion molecule (CD56) was constructed by glycosyl imprinting. A sandwich-like multi-signal generation strategy was first proposed in glycosyl imprinting sensors via boric acid affinity. Glycosyl-imprinted polymers were formed by electro-polymerization with poly-sialic acid (PolySia) as a template molecule and p-aminobenzeneboronic (p-ABA) acid as a functional monomer. Methods such as scanning electron microscope (SEM), Fourier transform infrared spectrum (FT-IR), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) were used to characterize the successful formation of imprinted membranes. Confirmed by both simulation calculation and experimental results, a signal-amplified effect based on macromolecules was introduced for the first time. After re-absorption, aminobenzene borate was linked to the surface of the sensor by boric acid affinity due to the rich hexadoxyl structure of the CD56-terminal chain as a signal probe. Under optimal conditions, the detection limit of the sensor is as low as 0.47 ng/L, and it can be successfully applied to the detection of CD56 in human serum.

Hippocampal overexpression of SGK1 ameliorates spatial memory, rescues Aß pathology and actin cytoskeleton polymerization in middle-aged APP/PS1 mice.

The increasing occurrence and ineffective treatment of Alzheimer's disease (AD) has become one of the major challenges of the world. Limited studies have shown that serum- and glucocorticoid-inducible kinase 1 (SGK1) is involved in spatial memory formation and consolidation, but its role in AD-like spatial memory impairment and the related mechanisms are not clear. In this study, we first examined the age-related changes of SGK1 in the hippocampus of female APP/PS1 (AD) mice. Based on the finding and our previous finding that significant spatial memory impairment was detected in 8-month old AD mice, SGK1-overexpressing AAV (oSGK1) was constructed and injected into the hippocampus of 9-month old AD mice. One month later, the behavior alterations, Aß production and deposit as well as changes of CA1 spine density and selected actin polymerization remodeling proteins were examined. The results showed that significant decrease of SGK1 was detected in 10-month old AD mice. The spatial memory impairment, the production and deposit of Aß were reversed by oSGK1. Levels of hippocampal ADAM10 (α-secretase) and IDE (Aß degradase), actin remodeling related proteins Rictor, Rac1, Cdc42 and Profilin-1 were significantly increased after oSGK1 treatment while hippocampal BACE1 (γ-secretase) and Cofilin remained unchanged. Taken together, our findings demonstrated a pivotal role of SGK1 in the treatment of AD-related memory impairment through upregulation of non- amyloidogenic processing of APP and degradation of Aß, increase in spine plasticity related proteins, indicating increase in hippocampal SGK1 may be a potent therapeutic target against AD.

TGF­ß1 upregulates the expression of lncRNA­ATB to promote atherosclerosis.

Transforming growth factor (TGF)­ß1 is reported to be associated with the occurrence of atherosclerosis, although the mechanism remains unclear. Therefore, the present study aimed to investigate the involvement of TGF­ß1 signaling in atherosclerosis. A total of 56 patients with atherosclerosis and 44 healthy volunteers were involved in this study. Serum expression of TGF­ß1 and long non­coding RNA­ATB was detected by ELISA and quantitative polymerase chain reaction (qPCR). Receiver operating characteristic curve analysis was performed to analyze the diagnostic value of serum TGF­ß1 and lncRNA­ATB for atherosclerosis. A human umbilical vein endothelial cell (HUVEC) line overexpressing lncRNA­ATB was constructed. The effects of TGF­ß1 treatment and lncRNA­ATB overexpression on HUVEC cell proliferation and viability was detected with Cell Counting Kit­8 and MTT assays, respectively. Expression of TGF­ß1 and pro­apoptotic Caspase­3 in lncRNA­ATB­overexpressing HUVECs was detected by western blotting. In addition, the expression of lncRNA­ATB in TGF­ß1­treated HUVECs was detected by qPCR. It was demonstrated that serum TGF­ß1 and lncRNA­ATB expression was significantly higher in atherosclerosis patients, compared with controls, and could be used to effectively distinguish patients from healthy individuals. TGF­ß1 treatment and lncRNA­ATB overexpression reduced HUVEC viability and proliferation. TGF­ß1 treatment increased the expression of lncRNA­ATB in HUVECs, while lncRNA­ATB overexpression had no significant effect on TGF­ß1 expression. LncRNA­ATB silencing with small interfering RNA significantly reduced the effects of TGF­ß1 treatment on the proliferation and viability of HUVECs. Furthermore, LncRNA­ATB overexpression upregulated the expression of caspase­3 in HUVECs. Therefore, it was concluded that TGF­ß1 may have upregulated the expression of lncRNA­ATB to promote atherosclerosis, and lncRNA­ATB may serve as a potential therapeutic target for atherosclerosis. However, the mechanism remains to be further investigated.

Age-related changes in hippocampal AD pathology, actin remodeling proteins and spatial memory behavior of male APP/PS1 mice.

Alzheimer's disease (AD) is the most common form of dementia in the elderly, characterized by amyloid-beta (Aß) plaques and tau neurofibrillary tangles (NFTs). Synaptic plasticity impairment is one of the early pathological events in AD. Transgenic APP/PS1 mice that overproduce Aß are one of the most extensively used AD animal models. Many studies have investigated the roles of NTF-related p-Tau, non-amyloidogenic ADAM10, amyloidogenic BACE1, Aß proteolytic NEP and IDE in certain ages of APP/PS1 mice as well as dendritic spine-related Rictor and Profilin-1 in normal mice, but there are few studies exploring the age-related changes of these molecules in the hippocampus of APP/PS1 mice. Furthermore, current studies regarding when memory impairment occurs in these mice are controversial. Thus, we examined the changes of these molecules in APP/PS1 and control mice using Western blot in mice 2-month-old (2 m) to 10 m of age and behavior changes using the Morris water maze from 4 m to 8 m. The results showed that in APP/PS1 mice, significant changes of hippocampal p-Tau, Aß, ADAM10, BACE1 and Rictor occurred at 6 m, NEP at 8 m, and IDE and Profilin-1 at 10 m. In control mice, changes of p-Tau, ADAM10, and BACE1 occurred at 8 m and NEP at 10 m, while IDE, Rictor and Profilin-1 remained unchanged. Importantly, the Morris water maze test revealed that spatial memory impairment was detected at 8 m but not 4 or 6 m. The above findings clearly evidence that neurochemical changes overtly precede cognitive dysfunctions in this AD model and provide novel knowledge for a better understanding of the molecular events driving AD.

GAG and collagen II attenuate glucocorticoid-induced osteoporosis by regulating NF-κB and MAPK signaling.

As a component of collagen II, glycosaminoglycan (GAG) has a relatively close relationship with bone metabolism. GAG and collagen II have been proven to promote connection of the bone trabecular structure. However, the exact mechanism remains unknown. In this study, we aimed to determine the concrete effect and the mechanism of GAG and collagen II on glucocorticoid-induced osteoporosis. We implanted prednisolone pellets subcutaneously in mice to mimic glucocorticoid-induced osteoporosis. GAG was administered intragastrically every day for 60 days. The results demonstrated a protective effect of GAG and collagen II on glucocorticoid-induced osteoporosis. Trabecular number and connection density increased after treatment with GAG and collagen II. We generated bone marrow-derived macrophages to explore the effect of GAG and collagen II on osteoclast differentiation. We collected cell protein and RNA in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator for nuclear factor-κB ligand (RANKL) and found that GAG and collagen II inhibited the NF-κB and MAPK pathways, thereby down-regulating osteoclast differentiation molecules such as matrix metallopeptidase 9 (MMP 9) and nuclear factor of activated T-cells, cytoplasmic 1 (NFATc-1). Our findings suggest that GAG and collagen II may have therapeutic potential of patients with glucocorticoid-induced osteoporosis in clinical settings.

Optimal FSH usage in revascularization of allotransplanted ovarian tissue in mice.

BACKGROUD: Ovarian transplantation is a useful method for preserving the fertility of young women with cancer who undergo radiotherapy and chemotherapy. Follicle-stimulating hormone (FSH) is use to protect transplanted ovarian tissues from ischemia injury through promoting revascularization after transplantation, but the side effect of high level FSH is ovarian overstimulation leading to substantial follicular loss. In this study, we investigated the optimal usage of FSH on revascularization in the in vitro cultured ovarian tissues before and after transplantation. RESULTS: FSH mainly exhibited an additive response in the gene and protein expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and follicle stimulating hormone receptor (FSHR) with its raised concentrations (0.15 IU/ml, 0.30 IU/ml and 0.60 IU/ml) and prolonged treatment (3 h, 6 h, 12 h, 24 h). The concentrations with 0.60 IU/ml FSH could obviously promoted the expression of VEGF, bFGF and FSHR, but under this concentration FSH could also overstimulated the ovarian tissue leading to follicular loss. With the increase of culture time, the gene and protein expression of VEGF and bFGF both were up-regulated in all of the FSH added groups, but FSHR expression decreased when culture time exceeded 12 h. So we chose 0.30 IU/ml FSH added concentration and 6 h culture time as the FSH usage condition in functional revascularization verification experiment, and found that under this condition FSH promoted 2.5 times increase of vascular density in treated group than in control group after ovarian tissues transplantation. CONCLUSION: Ovarian intervention with 0.30 IU/ml FSH for 6 h is an optimal FSH usage condition which could accelerate the revascularization in the allotransplanted ovarian tissue and can not produce ovarian overstimulation.