The GluN2B-Containing NMDA Receptor Alleviates Neuronal Apoptosis in Neonatal Hypoxic-Ischemic Encephalopathy by Activating PI3K-Akt-CREB Signaling Pathwa.

Neonatal hypoxic-ischemic encephalopathy (HIE) is a disease caused by insufficient blood supply in the brain in newborns during the perinatal period. Severe HIE leads to patient death, and patients with mild HIE are at increased risk of cognitive deficits and behavioral abnormalities. The NMDA receptor is an important excitatory receptor in the central nervous system, and in adult hypoxic-ischemic injury both subtypes of the NMDA receptor play important but distinct roles. The GluN2A-containing NMDA receptor (GluN2A-NMDAR) could activate neuronal protective signaling pathway, while the GluN2B-NMDAR subtype is coupled to the apoptosis-inducing signaling pathway and leads to neuronal death. However, the expression level of GluN2B is higher in newborns than in adults, while the expression of GluN2A is lower. Therefore, it is not clear whether the roles of different NMDA receptor subtypes in HIE are consistent with those in adults. We investigated this issue in this study and found that in HIE, GluN2B plays a protective role by mediating the protective pathway through binding with PSD95, which is quite different to that in adults. The results of this study provided new theoretical support for the clinical treatment of neonatal hypoxic ischemia.

Pancreatic Fat is not significantly correlated with ß-cell Dysfunction in Patients with new-onset Type 2 Diabetes Mellitus using quantitative Computed Tomography.

Objective: Type 2 diabetes mellitus (T2DM) is a chronic condition resulting from insulin resistance and insufficient ß-cell secretion, leading to improper glycaemic regulation. Previous studies have found that excessive fat deposits in organs such as the liver and muscle can cause insulin resistance through lipotoxicity that affects ß-cell function. The relationships between fat deposits in pancreatic tissue, the function of ß-cells, the method of visceral fat evaluation and T2DM have been sought by researchers. This study aims to elucidate the role of pancreatic fat deposits in the development of T2DM using quantitative computed tomography (QCT), especially their effects on islet ß-cell function. Methods: We examined 106 subjects at the onset of T2DM who had undergone abdominal QCT. Estimated pancreatic fat and liver fat were quantified using QCT and calculated. We analysed the correlations with Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) scores and other oral glucose tolerance test-derived parameters that reflect islet function. Furthermore, correlations of estimated pancreatic fat and liver fat with the area under the curve for insulin (AUCINS) and HOMA-IR were assessed with partial correlation analysis and demonstrated by scatter plots. Results: Associations were found between estimated liver fat and HOMA-IR, AUCINS, the modified ß-cell function index (MBCI) and Homeostatic Model Assessment ß (HOMA-ß). However, no significant differences existed between estimated pancreas fat and those parameters. Similarly, after adjustment for sex, age and body mass index, only estimated liver fat was correlated with HOMA-IR and AUCINS. Conclusions: This study suggests no significant correlation between pancreatic fat deposition and ß-cell dysfunction in the early stages of T2DM using QCT as a screening tool. The deposits of fat in the pancreas and the resulting lipotoxicity may play an important role in the late stage of islet cell function dysfunction as the course of T2DM progresses.

[A multicenter survey of the accessibility of essential medicines for children in China].

Objective: To investigate the availability, prices and affordability of essential medicines in pediatric population across China, in the hope of improving rational use of medicines. Methods: A multicenter cross-sectional survey of medicine prices, availability and affordability was conducted in 17 provinces, municipalities and autonomous region across east, south-central part, west and north of China. Data on 42 medicines used in pediatric population, both original and generic, were collected in 55 public hospitals from May 26 to June 2, 2017. Availability was expressed as the percentage of hospitals with stock of the target medicine on the day of data collection,and median price ratio (MPR) was the ratio of price upon investigation to international reference. Based on national minimum daily wage, affordability represents the number of working days needed to earn the expense which covers a standard course using the target medicine. Statistical software SPSS 13.0 was applied for descriptive analysis of availability, MPR and affordability. Results: Mean Availability of original and generic medicine was 33% and 32%, with median MPR being 5.43 and 1.55. Among the 19 medicines with price information for both original and generic product, the median MPR was 7.73 and 2.04 respectively. Regarding the five medicines used to treat four common pediatric diseases (pneumonia,peptic ulcer, congenital hypothyroidism, refractory nephrotic syndrome), the affordability was 0.63 (0.16-6.17) d for generic medicine, and 1.03 (0.16-11.53) d for its original counterpart. Conclusions: The availability to both original and generic products of the 42 medicines used in pediatric population was low in China. The prices of generic medicines seem to be lower and affordability higher than those of original medicines. There is an urgent need to improve the availability and affordability of pediatric medicines.

Association of von Willebrand factor (vWF) expression with lymph node metastasis and hemodynamics in papillary thyroid carcinoma.

OBJECTIVE: The purpose of this study was to analyze the relationship between von Willebrand factor (vWF) expression and lymph node metastasis or hemodynamics parameters in PTC. This work will provide a novel biomarker for the diagnosis of papillary thyroid carcinoma (PTC). PATIENTS AND METHODS: A total of 156 PTC patients were divided into metastatic and non-metastatic groups based on the presence or absence of lymph node metastasis. The Adler blood flow grading, color doppler flow imaging (CDFI), and blood flow index (PSV, PI, RI, AT) were measured and analyzed between the two groups. The expression of vWF was examined by immunocytochemical assay and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). The function of vWF was investigated by methyl thiazolyl tetrazolium (MTT) and the transwell assays. RESULTS: Both metastatic and non-metastatic groups with the major Adler grades as 0-1 had abundant blood flows. There was a significant difference in the rate of lymph node metastasis between Adler 2-3 and Adler 0-1. Moreover, the expression of vWF was found to be associated with lymph node metastasis or Adler blood flow grade in PTC. Significant differences in peak systolic velocity (PSV), systolic acceleration time (AT), and resistance index (RI) were detected in metastatic and non-metastatic groups. In addition, the upregulation of vWF was positively correlated with PSV, RI, and PI in PTC. Functionally, the knockdown of vWF inhibited the development of PTC by suppressing cell proliferation, migration, and invasion. CONCLUSIONS: Abnormal expression of vWF is closely related to lymph node metastasis and hemodynamics parameters in PTC patients. Furthermore, vWF plays an oncogene role in PTC progression.

Effectiveness of laser-assisted treatments for medication-related osteonecrosis of the jaw: a systematic review.

The feasibility of laser-assisted treatments of medication-related osteonecrosis of the jaw (MRONJ) remains poorly understood, so we have therefore systematically evaluated their effectiveness. We made a comprehensive search of MEDLINE, Pubmed, and Embase to find randomised controlled trials, case-control studies, and prospective cohort studies that assessed them. We assessed the eligible studies in duplicate, and if possible conducted a meta-analysis. Ten studies with a low to high risk of bias met the inclusion criteria. We found that a comparison of pain scores before and after using visible and infrared GaAs laser in the low-level laser treatment based on the Numerical Pain Rating Scale (mean difference 4.28; 95% CI 3.62 to 4.93; p<0.00001), showed that there were significant differences in the amount of pain. The effectiveness of other laser-assisted treatments on the reduction of pain - for example, Er:YAG laser surgical treatment, and laser-assisted treatment plus platelet-rich plasma, and the effect of other techniques on wound healing of laser-assisted treatments, are uncertain. We found that the results of the studies that were deemed to be high-to-low quality and to have high-to-low statistical power suggested that there may be considerable clinical improvement in MRONJ by using laser-assisted treatment; we cautiously consider that low-level laser treatment may manage pain and symptoms in these patients. More randomised studies of good quality and with a low risk of bias are needed to test whether laser-assisted treatment should be a routine part of management of patients with MRONJ.

[The progress and prospect of ophthalmological research on cerebral visual impairment in children].

With the rapid development of global perinatal medicine and neonatal medicine, the survival rate of high-risk neonates (premature, and those who suffer neurologic lesion during or shortly after birth, et al.) improved significantly, however the incidence of cerebral visual impairment (CVI) also rose, which has become the most common cause of visual impairment in children in developed countries. Studies found that visual abnormalities of patients with CVI can be various. Since children's cognition and motor development are inseparable from visual functions, children with CVI are usually characterized by abnormalities in sensory perception, cognition and even movement, other than visual impairment itself. Due to the characteristics of CVI, such as early onset, complex etiology, difficulty in diagnosis and treatment, and involvement with ophthalmology, pediatrics, rehabilitation medicine, genetic epidemiology and other multi-disciplinary content, current domestic research on CVI is limited. From the perspective of ophthalmologist, this paper reviews the progress of ophthalmology diagnosis and treatment in children with CVI in recent years, aiming to have better early recognition and individualized intervention, so as to help pediatrician and rehabilitation physicians to improve survival skills for CVI children and their quality of life. (Chin J Ophthalmol, 2019, 55:469-474).

Cytological mechanism of astragaloside IV in promoting repair of bone defects.

To explore the possible cytological mechanism underlying the role of Astragaloside IV in promoting the repair of bone defects, osteoblasts were cultured in vitro and identified using inverted phase contrast microscopy, alkaline phosphatase (ALP) staining and alizarin red staining.

[Facial symmetry after conservative treatment of unilateral condylar fracture in children: a three-dimensional study].

Objective: To investigate the influence of condylar fractures on the growth of condylars after conservative treatments by three-dimensional measurement. Methods: Twenty three children with unilateral condylar fracture followed-up for at least 1 year in Shanghai Ninth People's Hospital were included from June 2016 to March 2017. CT scans were performed for each patient. Three-dimensional virtual skulls were reconstructed and measuring points were defined with Simplant 11.04 software. The depth of glenoid fossa, height of articular eminence, width, height and depth of condylar, deviation of pogonion were measured. Statistical analyses were conducted to assess difference between the bifid group and the normal group. Results: No difference was observed in the vertical position of crest of the articular eminence between two groups (P=0.110). The vertical position of roof of the glenoid fossa in bifid side was significantly superior than the one in normal side (P=0.010). Bifid side had a shorter (P=0.002) and wider(P=0.002) condylar than normal side did. No difference was observed in the depth of condylar between two groups (P=0.071). The average deviation of pogonion was (0.69±1.75) mm (P=0.072) from sagittal plan. Conclusions: This short-termed study indicates that children's growth potential of condylar is under average after conservative treatments. Because the bone hyperosteogeny of glenoid fossa compensates the hypotrophy of condylar, no deviation of pogonion is detected.

[Prognostic influence of preoperative Nutritional Risk Screening -2002 (NRS-2002) score for patients with thoracic esophageal squamous cell carcinoma receiving surgery].

Objective: To apply Nutritional Risk Screening-2002(NRS-2002) to perform nutritional status score for the patients with thoracic esophageal squamous cell carcinoma (ESCC) receiving surgery, and to explore the prognostic impact of long-term survival. Methods: A total of 117 patients who were diagnosed with ESCC from 2010 to 2012 were retrospectively analyzed. They recieved standard curative esophagectomy in the Yangzhou University Affiliated Taixing People's Hospital. The nutritional status and risk score for recruited patients were assessed according to the standard of NRS-2002 tool prior to surgery, and these patients were grouped for further analysis according to the median values of NRS-2002 score. Finally, the relationship between NRS-2002 score and prognosis was analyzed. Results: Patients were classified into two groups, with 45 in the NRS-2002<2.0 group, and 72 cases in the NRS-2002≥2.0, respectively. In the NRS-2002<2.0 group, the 1-, 3-, and 5-year progression-free survival (PFS) rates were 75.6%, 44.4% and 40.0% separately, while in the NRS-2002≥2.0 group, the PFS rates were 61.1%, 6.9% and 4.2% respectively, and the differences were statistically significant (P<0.001). Correspondingly, in the NRS-2002< 2.0 group, the 1-, 3-, and 5-year overall survival (OS) rates were 97.8%, 66.7% and 57.8% separately, while in the NRS-2002≥2.0 group, the OS rates were 91.7%, 33.3% and 16.7% respectively, and the differences were also statistically significant (P<0.001). Univariate analysis showed that N stage, TNM stage and NRS-2002 score were closely related to PFS and OS (P<0.05), and T stage was only associated with OS in patients with thoracic esophageal squamous cell carcinoma (P<0.05). Furthermore, multivariate Cox regression analysis showed that N stage (RR=1.640, 95% CI 1.049-2.565, P=0.030) and NRS-2002 (RR=3.154, 95% CI 1.946-5.113, P<0.001) were independent prognostic factors for PFS in patients with ESCC after surgery. Additionally, pathological differentiation (RR=1.556, 95% CI 1.004-2.440, P=0.041), N stage (RR=1.624, 95% CI 1.017-2.593, P=0.042) and NRS-2002 (RR=3.906, 95% CI2.245-6.795, P<0.001) were independent prognostic factors for OS in ESCC patients following surgery. Conclusion: Preoperative nutritional risk screening NRS-2002 score is an independent prognostic factor in patients with ESCC receiving surgery and could be used as a tool for primary screening for nutritional risk.

Variability in carbon uptake and (re)cycling in Antarctic cryptoendolithic microbial ecosystems demonstrated through radiocarbon analysis of organic biomarkers.

Cryptoendolithic lichens and cyanobacteria living in porous sandstone in the high-elevation McMurdo Dry Valleys are purported to be among the slowest growing organisms on Earth with cycles of death and regrowth on the order of 103 -104  years. Here, organic biomarker and radiocarbon analysis were used to better constrain ages and carbon sources of cryptoendoliths in University Valley (UV; 1,800 m.a.s.l) and neighboring Farnell Valley (FV; 1,700 m.a.s.l). Δ14 C was measured for membrane component phospholipid fatty acids (PLFA) and glycolipid fatty acids, as well as for total organic carbon (TOC). PLFA concentrations indicated viable cells comprised a minor (<0.5%) component of TOC. TOC Δ14 C values ranged from -272‰ to -185‰ equivalent to calibrated ages of 1,100-2,550 years old. These ages may be the result of fractional preservation of biogenic carbon and/or sudden large-scale community death and extended period(s) of inactivity prior to slow recolonization and incorporation of 14 C-depleted fossil material. PLFA Δ14 C values were generally more modern than the corresponding TOC and varied widely between sites; the FV PLFA Δ14 C value (+40‰) was consistent with modern atmospheric CO2 , while UV values ranged from -199‰ to -79‰ (calibrated ages of 1,665-610 years). The observed variability in PLFA Δ14 C depletions is hypothesized to reflect variations in the extent of fixation of modern atmospheric CO2 and the preservation and recycling of older organic carbon by the community in various stages of sandstone recolonization. PLFA profiles and microbial community compositions as determined by molecular genetic characterizations and microscopy differed between the two valleys (e.g., predominance of biomarker 18:2 [>50%] in FV compared to UV), representing microbial communities that may reflect distinct stages of sandstone recolonization and/or environmental conditions. It is thus proposed that Dry Valley cryptoendolithic microbial communities are faster growing than previously estimated.

Functional roles of exosomes in cardiovascular disorders: a systematic review.

Cardiovascular diseases are major causes of people death associated with high mortality and disability. Exosomes are nanosized extracellular vesicles containing protein, lipid, transcription factors, mRNAs, non-coding RNA (ncRNA) and nucleic acid contents, which are critical players of intercellular communication via long-range signals or cell-to-cell contact. The emergence of exosomes provides favorable strategies for the diagnosis and treatment of cardiovascular diseases. Exosomes-based molecular mechanisms are important for developing novel therapeutic approaches for cardiovascular events. In this review, we will (1) provide insights into the detrimental and beneficial effects of exosomes on cardiovascular physiology, (2) summarize the underlying biological mechanisms of the exosome in cardiovascular events, (3) investigate the therapeutic value of exosomes for cardiovascular disorders.

Chymotrypsin with sialendoscopy-assisted surgery for the treatment of chronic obstructive parotitis.

Chronic obstructive parotitis (COP) is a common disease of the parotid gland. A total of 104 patients with COP were identified and randomized into a treatment group (52 cases) and a control group (52 cases). All patients underwent sialography and salivary gland scintigraphy (SGS) examinations before surgery. The patients in the treatment group received chymotrypsin combined with gentamicin via interventional sialendoscopy to irrigate the duct, and the control group received gentamicin alone. All patients were asked to record their pain on a visual analogue scale (VAS) before treatment and at 1 week, 2 weeks, 1 month, 3 months, and 6 months after surgery. The VAS score for pain intensity was decreased at 1 week post-treatment in both groups (P<0.05). Compared to the control group, the VAS score was lower in the treatment group at 1 week, 2 weeks, and 1 month post-treatment (P<0.05). The 6-month postoperative SGS results showed improved uptake and excretion in both groups (P<0.05). The treatment group exhibited higher scores for postoperative SGS excretion than the control group (P<0.05). The administration of chymotrypsin combined with gentamicin by sialendoscopy is effective for the treatment of non-stone-related COP and specifically improves the excretion function of the parotid gland.

Centrally mediated ejaculatory response via sympathetic outflow in rats: role of N-methyl-D-aspartic acid receptors in paraventricular nucleus.

Ejaculation is mediated by a spinal generator, which integrates inputs related to the sexual activity and coordinates sympathetic, parasympathetic, and motor outflow. Previous clinical studies indicate that primary premature ejaculation is related to the hyperactivity of the sympathetic nervous system. In this study, we explored the roles of N-methyl-D-aspartic acid (NMDA) receptors in paraventricular nucleus of the hypothalamus (PVN) on ejaculatory responses and its potential mechanism in the rats. We found that microinjection of 0.20 nmol NMDA into the PVN reduced the latency of intromission and facilitated ejaculation during copulation. Moreover, delayed ejaculation and intromission were observed after the rats were microinjected with NMDA receptor antagonist D (-)-2-Amino-5-phosphonopentanoic acid (AP-5). However, we discovered that microinjection of NMDA into PVN significantly increased baseline lumbar splanchnic nerve activity (LSNA), and the NMDA dose was positively correlated with the increased LSNA (r = 0.875, p = 0.04). Meanwhile, the plasma norepinephrine level in rats injected with NMDA was much higher than that in rats injected with saline (1453.4 ± 136.4 pg/mL vs. 492.3 ± 36.8 pg/mL, p < 0.01). Additionally, AP-5 reduced the baseline LSNA and abrogated the enhancing activity of NMDA in LSNA. Thus, we propose that NMDA receptors in PVN may facilitate ejaculation through enhancing the activity of sympathetic system.

[Medial anterior flange overhang related to the prosthesis selection determined by femoral sizer in total knee arthroplasty].

Objective: To evaluate the medial anterior flange overhang related to the femoral prosthesis size determined by femoral sizer in total knee arthroplasty. Methods: Sixty one cases of varus knee osteoarthritis (61 knees) were analyzed from January 2014 to January 2015, 14 males and 47 females, aged (66.8±6.8) year old. 3D models of distal femur were established by Mimics, total knee arthroplasty surgeries were simulated and the medial and lateral femoral condylar anteroposterior diameter were measured. According to the AP diameter of lateral femoral condyle (lateral condyle group) and femoral sizer (sizer group), different size of the PFC Sigma femoral prosthesis models were imported to mimics, and the coverage of medial anterior femoral flange were compared. Results: The AP diameter of lateral femoral condyle[(55.76±3.64) mm]was significantly less than the sizer group[(57.02±3.59) mm], P<0.05. The distal femur aspect ratio for male (1.2 %±0.1%) and female (1.2%±0.1%) has no statistically difference (P>0.05). Femoral prosthesis sizes, the uncoverage diameter of the anterior chamfer line, midpoint of the flange in the sizer group were significantly larger than the lateral condyle group (P<0.05). The femoral size, overhang ratio of anterior chamfer line and midpoint of the flange in male were significantly larger than females (P<0.05). Conclusion: The AP diameter determined by femoral sizer instrument had a tendency to larger prosthesis, and the risks of medial femoral anterior flange overhang increased. Medial anterior flange overhang in male was significantly less than female, but the difference had no correlation to the distal femur asepect ratio.

Superoxide anions in the paraventricular nucleus mediate cardiac sympathetic afferent reflex in insulin resistance rats.

AIM: Cardiac sympathetic afferent reflex (CSAR) participates in sympathetic over-excitation. Superoxide anions and angiotensin II (Ang II) mechanisms are associated with sympathetic outflow and CSAR in the paraventricular nucleus (PVN). This study was designed to investigate whether PVN superoxide anions mediate CSAR and Ang II-induced CSAR enhancement response in fructose-induced insulin resistance (IR) rats. METHODS: CSAR was evaluated with the changes of renal sympathetic nerve activity (RSNA) and mean arterial pressure (MAP) responses to the epicardial application of capsaicin (CAP) in anaesthetized rats. RESULTS: Compared with Control rats, IR rats showed that CSAR, PVN NAD(P)H oxidase activity, superoxide anions, malondialdehyde (MDA), Ang II and AT1 receptor levels were significantly increased, whereas PVN superoxide dismutase (SOD) and catalase (CAT) activities were decreased. In Control and IR rats, PVN microinjection of superoxide anions scavengers tempol, tiron and PEG-SOD (an analogue of endogenous superoxide dismutase) or inhibition of PVN NAD(P)H oxidase with apocynin caused significant reduction of CSAR, respectively, but DETC (a superoxide dismutase inhibitor) strengthened the CSAR. PVN pre-treatment with tempol abolished, whereas DETC potentiated, Ang II-induced CSAR enhancement response. Moreover, PVN pre-treatment with tempol or losartan prevented superoxide anions increase caused by Ang II in IR rats. CONCLUSION: PVN superoxide anions mediate CSAR and Ang II-induced CSAR response in IR rats. In IR state, increased NAD(P)H oxidase activity and decreased SOD and CAT activities in the PVN promote superoxide anions increase to involve in CSAR enhancement. Ang II may increase NAD(P)H oxidase activity via AT1 receptor to induce superoxide anion production.

Apelin-13 and APJ in paraventricular nucleus contribute to hypertension via sympathetic activation and vasopressin release in spontaneously hypertensive rats.

AIMS: Apelin is a specific endogenous ligand of orphan G protein-coupled receptor APJ. This study was designed to determine the roles and mechanisms of apelin-13 and APJ in paraventricular nucleus (PVN) in renal sympathetic nerve activity (RSNA), arginine vasopressin (AVP) release and mean arterial pressure (MAP) in spontaneously hypertensive rats (SHR). METHOD: Acute experiment was carried out in 13-week-old male SHR and Wistar-Kyoto rats (WKY) under anaesthesia. RSNA and MAP responses to the PVN microinjection were determined. Apelin and APJ expressions were examined with quantitative real-time PCR and Western blot. AVP and noradrenaline were determined with ELISA. Osmotic minipumps were used for chronic PVN infusion in conscious WKY. RESULTS: Apelin and APJ in the PVN were up-regulated in SHR. The PVN microinjection of apelin-13 increased, but APJ antagonist F13A decreased the RSNA, MAP, plasma noradrenaline and AVP levels in SHR. N-methyl-D-aspartate receptor (NMDAR) antagonist plus non-NMDAR antagonist abolished the apelin-13-induced sympathetic activation rather than AVP release. NMDAR antagonist or non-NMDAR antagonist alone attenuated the apelin-13-induced sympathetic activation. Chronic infusion of apelin-13 into the PVN in normotensive rats induced hypertension, increased plasma noradrenaline and AVP levels and promoted myocardial atrial natriuretic peptide and beta-myosin heavy chain mRNA expressions, two indicative markers of cardiac hypertrophy. CONCLUSION: Apelin-13 and APJ in the PVN contribute to hypertension via sympathetic activation and AVP release in SHR. The sympatho-excitatory effect of apeline-13 is mediated by both NMDAR and non-NMDAR in the PVN. Persistent activation of APJ in the PVN induces hypertension.

Morphological characteristics of the walls of thrombophlebitic saphenous vein.

INTRODUCTION: To investigate the morphological changes in the walls of thrombophlebitic saphenous veins. METHODS: Fifty-four specimens were made into slices for haematoxylin and eosin and Masson trichrome staining; ultrathin slices were also created. Slices were observed under light microscopy and electron microscopy. RESULTS: Under light microscopy, the tunica intima of venous wall in the thrombophlebitic saphenous veins group was obviously thickened and incomplete; intravascular papillary endothelial hyperplasia was observed. Hyperplasia of collagenous fibres in the tunica media was observed. Elastic fibres in the tunica externa became fewer, whereas nourishing vessels were significantly increased in number. Infiltration of many inflammatory cells was observed. Under electron microscopy, Auer bodies with high electron density and round granules could be seen in endothelial cells in the thrombophlebitic saphenous veins group. Smooth muscle cells had an irregular karyotype, with blurred cristae in some mitochondria. Myofilaments basically disappeared. CONCLUSION: Thrombus formation might aggravate re-modelling of the walls of varicose veins.

Transcriptome sequencing of sea cucumber (Apostichopus japonicus) and the identification of gene-associated markers.

Sea cucumber (Apostichopus japonicus) is an ecologically and economically important species in East and South-East Asia. This project aimed to identify large numbers of gene-associated markers and differentially expressed genes (DEGs) after lipopolysaccharides (LPS) challenge in A. japonicus using high-throughput transcriptome sequencing. A total of 162 million high-quality reads of 174 million raw reads were obtained by deep sequencing using Illumina HiSeq™ 2000 platform. Assembly of these reads generated 94 704 unigenes, with read length ranging from 200 to 16 153 bp (average length of 810 bp). A total of 36 005 were identified as coding sequences (CDSs), 32 479 of which were successfully annotated. Based on the assembly transcriptome, we identified 142 511 high-quality single nucleotide polymorphisms (SNPs). Among them, 33 775, 63 120 and 45 616 were located in sequences without predicted CDS (non-CDSs), CDSs and untranslated regions (UTRs), respectively. These putative SNPs included 82 664 transitions and 59 847 transversions. Totally, 89 375 (59.1%) were distributed in 15 473 known genes. A total of 6417 microsatellites were detected in 5970 unigenes, 3216 of which were annotated and 2481 were successfully subjected for primer design. The numbers of simple sequence repeats (SSRs) identified in non-CDSs, CDSs and UTRs were 2367, 2316 and 1734. These potential SNPs and SSRs are expected to provide abundant resources for genetic, evolutionary and ecological studies in sea cucumber. Transcriptome comparison revealed 1330, 1347 and 1291 DEGs in the coelomocytes of A. japonicus at 4 h, 24 h and 72 h after LPS challenge, respectively. Approximately 58.4% (1802) of total DEGs have been successfully annotated.

Superoxide anions involved in sympathoexcitation and pressor effects of salusin-ß in paraventricular nucleus in hypertensive rats.

AIMS: Salusin-ß in paraventricular nucleus (PVN) increases renal sympathetic nerve activity (RSNA), mean arterial pressure (MAP), heart rate (HR) and arginine vasopressin (AVP) release in hypertensive rats but not in normal rats. The present study was designed to investigate the downstream molecular mechanism of salusin-ß in the PVN in hypertension. METHOD: Renovascular hypertension was induced by two-kidney, one-clip (2K1C) in male SD rats. Acute experiments were carried out 4 weeks after 2K1C or sham operation under anaesthesia. RESULTS: MrgA1 mRNA expression and salusin-ß level in the PVN as well as plasma salusin-ß level were increased in 2K1C rats. Bilateral PVN microinjection of salusin-ß increased the RSNA, MAP and HR in 2K1C rats, which were abolished by the pre-treatment with polyethylene glycol-superoxide dismutase (PEG-SOD), the superoxide anion scavenger tempol, the NAD(P)H oxidase inhibitor apocynin or the protein kinase C (PKC) inhibitor chelerythrine chloride (CLC), but not affected by the AT1 receptor antagonist losartan, the Mas receptor antagonist A-779, the NOS inhibitor L-NAME or the GABAA and GABAB receptor antagonists gabazine+CGP-35348. Salusin-ß-induced increases in superoxide anion level and NAD(P)H oxidase activity in the PVN were abolished by the PVN pre-treatment with CLC. Salusin-ß increased AVP levels in rostral ventrolateral medulla and plasma, which were prevented by the pre-treatment with PEG-SOD, apocynin or CLC in 2K1C rats. Salusin-ß augmented the enhanced activity of PKC in the PVN in 2K1C rats. CONCLUSION: Protein kinase C-NAD(P)H oxidase-superoxide anions pathway in the PVN is involved in salusin-ß-induced sympathetic activation, pressor response and AVP release in renovascular hypertension.