Gliomas are the most common primary intracranial neoplasms among all brain malignancies, and the microtubule affinity regulating kinases (MARKs) have become potential drug targets for glioma. Here, we report a novel dual small-molecule inhibitor of MARK3 and MARK4, designated as PCC0208017. PCC0208017 strongly inhibited kinase activity against MARK3 and MARK4, and strongly reduced proliferation in three glioma cell lines. This compound attenuated glioma cell migration, glioma cell invasion, and angiogenesis. Molecular mechanism studies revealed that PCC0208017 decreased the phosphorylation of Tau, disrupted microtubule dynamics, and induced a G2/M phase cell cycle arrest. In an glioma model, PCC0208017 showed robust anti-tumor activity, blood-brain barrier permeability, and a good oral pharmacokinetic profile. Molecular docking studies showed that PCC0208017 exhibited high binding affinity to MARK3 and MARK4. Taken together, our study describes for the first time that PCC0208017, a novel MARK3/MARK4 inhibitor, might be a promising lead compound for treatment of glioma.

Objective To investigate the effects of matrine on proliferation and apoptosis of human Tenon capsule fibroblasts (HTFs) in.vitro.Methods After treated with 0,0.3,0.6 and 0.9 g/L matrine in vitro,cell counting kit-8 (CCK-8) method was used to assay the proliferation of HTFs at 24,48 and 72 hours,Western blot and PCR were performed to evaluate the expression of apoptosis-associated factor caspase-3 on both protein and RNA levels.Results The activity of human Tenon capsule fibroblast at 48 hours and 72 hours after treated with 0.3,0.6,0.9 g/L matrine was significantly inhibited when compared with the 0 g/L matrine group,and the inhibitory effect was dose-dependent and time-dependent (F ion =1 019.51,P =0.00;Ftime =5 848.66,P =0.00;Fi ion =147.45,P=0.00).After treated with 0,0.3,0.6 and 0.9 g/L matrine,the early apoptosis rate of HTFs was (2.68±0.30)％,(5.08±0.47)％,(6.97±0.69)％ and (10.30±1.20)％,the grey value ofcaspase-3 protein was 1.00±0.13,1.90±0.19,2.50±0.30 and 2.67±0.30,the relative expression of caspase-3 mRNA was 0.98 ±0.12,2.01 ±0.34,6.15 ± 0.60 and 11.40 ± 1.12,respectively,with significant differences among them (F =55.74,66.01,154.50;all at P＜0.01),the early apoptosis rate of HTFs,the grey value of caspase-3 protein and the relative expression of caspase-3 mRNA were all increased significantly as the concentration of matrine increased,with significant differences between any two groups (all at P＜0.05).Conclusions Matrine can inhibit the proliferation of HTFs and induce the apoptosis of HTFs in a time-and dose-depended manner.

Objective To investigate whether mild hypothermia can promote neurogenesis in the dentate gyrus of hippocampus and cognitive function recovery after traumatic brain injury ( TBI) through inhibiting apoptosis of hippocampal neurons. Methods A total of 66 healthy adult Sprague-Dawley rats were randomly divided into sham group, TBI group and TBI+hypothermia group, with 22 rats in each group. The rat TBI model was established using the fluid percussion device. The rats in TBI +hypothermia group received 4-hour hypothermia therapy immediately after injury, with the target temperature of 33. 5℃. Bromodeoxyuridine (BrdU) was injected into the rats' abdominal cavity to label the mitotic cells. The test of Morris water maze was used to evaluate the rats' spatial learning and memory capabilities. Immunofluorescence staining was used to observe the expression levels of BrdU, doublecortin (DCX), neuron specific nuclear protein (NeuN), cysteinyl aspartate specific proteinase 3 (caspase-3) and cleaved caspase-3 expressions in dentate gyrus of hippocampus at 7 days and 28 days after injury. Expressions apoptosis-related proteins including the factor associated suicide ( FAS )/factor associated suicide ligand (FASL), B-cell lymphoma-2 (Bcl-2), caspase-3 and cleaved caspase-3 expressions were detected by Western blot assay. Results The water maze tests at 28 days after injury showed that compared with TBI group, the escape latency in TBI+hypothermia group was significantly shorter [(24. 2 ± 5. 9)s:(18 ± 4. 1)s], and both the time in the target quadrant and the number of platform crossing were increasedsignificantly[(24.9±6.5)s:(31.7±5.2)s; (1.9±0.8) times:(3.5±1.2)times](P<0. 05). Compared with the sham group, in TBI group and TBI+hypothermia group, the BrdU+ new-born cells in the dentate gyrus of hippocampus were significantly increased at 7 days after injury [(9. 4 ± 4. 1):(33. 4 ± 3. 8);(9. 4 ± 4. 1):(45. 8 ± 5. 6)], the BrdU+ /DCX+ new-born neurons were increased at 7 days after injury [(2. 0 ± 0. 6):(9. 6 ± 1. 6);(2. 0 ± 0. 6):(19. 2 ± 3. 7)], and the BrdU+ /NeuN+mature neurons were increased at 28 days after injury [(2. 6 ± 1. 0) :(17. 2 ± 3. 9); (2. 6 ± 1. 0) :(33. 6 ± 9. 1)] (P<0. 01). TBI group showed more obvious increase than the TBI+hypothermia group (P<0. 01). Moreover, compared with 7 days after injury, the number of BrdU+ cells at 28 days after injury was further increased in TBI +hypothermia group but decreased in TBI group [(45. 8 ± 5. 6) :(58. 8 ± 9. 2);(33. 4 ± 3. 8):(22. 0 ± 3. 5)](P<0. 05 or <0. 01). Compared with the sham group, the caspase-3 +NeuN+ and caspase-3 +NeuN+ apoptotic neurons were significantly increased at 7 days after injury in TBI group [(2. 0 ± 0. 9):(11. 6 ± 2. 6); (2. 6 ± 1. 0):(10. 2 ± 2. 9)] (P<0. 05). Compared with the TBI group, the cleaved caspase-3 +NeuN+ apoptotic neurons were decreased in TBI+hypothermia group [(6. 6 ± 2. 0):(11. 6 ± 2. 6)](P<0. 05). Furthermore, compared with the TBI group, mild hypothermia might down-regulate the expression of FAS, FASL, cleaved caspase-3 and caspase-3 and up-regulate the expression of Bcl-2 in the hippocampus [(1. 54 ± 0. 15) :(1. 14 ± 0. 12);(1. 06 ± 0. 04):(0. 80 ± 0. 09); (0. 84 ± 0. 03):(0. 62 ± 0. 08); (0. 93 ± 0. 06):(0. 86 ± 0. 09);(0. 71 ± 0. 01):(1. 58 ± 0. 18)](P<0. 05). Conclusions Mild hypothermia might inhibit apoptosis of hippocampal neurons through cleaved caspase-3, FAS/FASL and Bcl-2 pathways, thus improving the neurogenesis and maturation of neurons in the dentate gyrus of hippocampus and facilitating cognitive function recovery in rats. It indicates that the function of hypothermia in anti-apoptosis and neurogenesis and maturity of hippocampal neurons may have a potential role in predicting the prognosis of TBI patients.

The aim of this study is to investigate the effect of long non-coding RNA PLUTO on β cell function and its underlying mechanism. Islet cells of db/db, ob/ob and HFD mice were isolated and extracted by enzymatic method to detect the expression of PLUTO in islet cells of different obesity model mice. Glucolipid toxicity was used to stimulate islet primary cells and Min6 cells, and the expression of PLUTO was detected to elucidate the reasons of PLUTO down-regulation induced by obesity factors. RT-qPCR and ELISA were performed to analyze the function of PLUTO on β cells; Western blot and rescue experiments were carried out to elucidate the regulatory mechanism of β cells by PLUTO. The results showed that the expression levels of PLUTO were significantly decreased in obesity model mice compared with control mice; over-expression PLUTO in primary islets and Min6 cells could improve insulin biosynthesis and secretion; Western blot analysis showed that PLUTO promoted insulin secretion and synthesis by up-regulating the transcription and translation of Pdx1, which is the adjacent gene of PLUTO.

Objective To observe the effect of mild hypothermia on myocardial β-adrenergic receptor (β-AR) signal pathway after cardiopulmonary resuscitation (CPR) in pigs with cardiac arrest (CA) and explore the mechanism of myocardial protection. Methods Healthy male Landraces were collected for reproducing the CA-CPR model (after 8-minute untreated ventricular fibrillation, CPR was implemented). The animals were divided into two groups according to random number table (n = 8). In the mild hypothermia group, the blood temperature of the animals was induced to 33 ℃ and maintained for 6 hours within 20 minutes after return of spontaneous circulation (ROSC) by using a hypothermia therapeutic apparatus. In the control group, the body temperature of the animals was maintained at (38.0±0.5)℃ with cold and warm blankets. The heart rate (HR), mean arterial pressure (MAP), the maximum rate of increase or decrease in left rentricular pressure (+dp/dt max)were measured during the course of the experiment. The cardiac output (CO) was measured by heat dilution methods before CA (baseline), and 0.5, 1, 3, 6 hours after ROSC respectively, the venous blood was collected to detect the concentration of cTnI. Left ventricular ejection fraction (LVEF) was measured with cardiac ultrasound before CA and 6 hours after ROSC. Animals were sacrificed at 6 hours after ROSC and the myocardial tissue was harvested quickly, the mRNA expression of β1-AR in myocardium was detected by reverse transcription-polymerase chain reaction (RT-PCR), the contents of adenylate cyclase (AC) and cyclic adenosine monophosphate (cAMP) were detected by enzyme linked immunosorbent assay (ELISA), the protein content of G protein-coupled receptor kinase 2 (GRK2) was detected by Western Blot. Results After successful resuscitation, the HR of both groups were significantly higher than the baseline values, CO, ±dp/dt max were significantly decreased, MAP were not significantly changed, serum cTnI levels were significantly increased. Compared with the control group, HR at 0.5, 1, 3 hours after ROSC were significantly decreased in mild hypothermia group (bpm: 142.80±12.83 vs. 176.88±15.14, 115.80±11.48 vs. 147.88±18.53, 112.60±7.40 vs. 138.50±12.02, all 1 < 0.01), CO was significantly increased at 1 hours and 3 hours after ROSC (L/min: 3.97±0.40 vs. 3.02±0.32, 4.00±0.11 vs. 3.11±0.59, both 1 < 0.01), +dp/dt max at 3 hours and 6 hours was also significantly increased after ROSC [+dp/dt max (mmHg/s): 3 402.5±612.7 vs. 2 130.0±450.6, 3 857.5±510.4 vs. 2 562.5±633.9; -dp/dt max (mmHg/s): 2 935.0±753.2 vs. 1 732.5±513.6, 3 520.0±563.6 vs. 2 510.0±554.3, all 1 < 0.05], the cTnI was significantly decreased at 3 hours and 6 hours afher ROSC (μg/L: 1.39±0.40 vs. 3.24±0.78, 1.46±0.35 vs. 3.78±0.93, both 1 < 0.01). The left at 6 hours after ROSC in both groups was decreased as compared with that before CA. The LVEF in the mild hypothermia group was higher than that in the control group (0.52±0.04 vs. 0.40±0.05, 1 < 0.05). The mRNA expression of β1-AR, and concentrations of AC and cAMP in hypothermia group were significantly higher than those in control group [β1-AR mRNA (2-ΔΔCT): 1.18±0.39 vs. 0.55±0.17, AC (ng/L):197.0±10.5 vs. 162.0±6.3, cAMP (nmol/L): 1 310.58±48.82 vs. 891.25±64.95, all 1 < 0.05], GRK2 was lower than that in the control group (GRK2/GAPDH: 0.45±0.05 vs. 0.80±0.08, 1 < 0.05). Conclusion Mild hypothermia can reduce the degree of cardiac function injury after CPR, and its mechanism may be related to the reduction of impaired myocardial β-AR signaling after CPR.

Objective To investigate the protective effect and mechanism of serum containing Euonymus fortunei on the rat pancreatic islet cells. Methods Forty male SD rats were randomly divided into 5 groups (n=8 in each group), including the control group (normal rat islet cells were cultured with normal rat serum), ischemic preconditioning group (abdominal aorta was blocked first and then re-opened before the pancreas was obtained, and the pancreatic islet cells were cultured with normal rat serum), Euonymus fortunei treatment group (normal rat islet cells were cultured with rat serum containing Euonymus fortunei), Euonymus fortunei group and blank group (normal rats were administered orally with Euonymus fortunei extract or distilled water for the preparation of rat serum). Diphenylthiocarbazone (DTZ) staining was utilized to observe and calculate the quantity of islets. Acridine orange (AO)/propidium iodide (PI) staining was adopted to calculate the survival rate of islet cells. The insulin release experiment was performed to calculate the stimulation index (SI) and evaluate islet cell function. The concentration of glutathione (GSH) and nitric oxide (NO) in islet cells was detected using GSH and NO kits. The expression level of inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). Results Islet cells were observed in specifically scarlet color after DTZ staining. The quantity of islet cells did not significantly differ among different groups (all P>0.05). Along with the prolongation of culture time, the activity of islet cells in each group was gradually decreased. At 72 h after isolation and culture, compared with the control group, the survival rate of the cells was significantly higher in the Euonymus fortunei treatment group (P<0.05). The insulin release test results demonstrated that compared with the control group, the SI of the ischemic preconditioning and Euonymus fortunei treatment groups was significantly increased (both P<0.05). Compared with the control group, the GSH contents of pancreatic islet cells in the ischemic preconditioning and Euonymus fortunei treatment groups were considerably enhanced, the NO content was significantly decreased, and the expression level of iNOS mRNA was significantly down-regulated (all P<0.05). Conclusions Euonymus fortunei can increase the survival rate of islet cells and enhance the function of pancreatic islets by increasing the level of GSH, down-regulating the expression of iNOS and decreasing the NO production.

Objective To investigate the characteristic of lipoprotein(a)[Lp(a)]in different phases of chronic kidney disease (CKD ),to provide the basis for clinical prevention and treatment of CKD.Methods 200 patients with CKD in the Republic Hospital of Shifang were collected as study group,including 5 phases (every phase had 40 cases),and 100 healthy people were selected as control group.Measured the serum Lp(a)of both study and control group,analyzed the correlations between Lp(a)and different phase of CKD.All data were analyzed by SPSS version 17.0.The significant level was established at 0.05.Results CKD1 [(146.0 ±95.5)mg/L]and all CKD group [(231.5 ±133.2)mg/L]had higher level of serum Lp(a)than the control group [(115.5 ±70.2)mg/L] (Z=-2.800,P<0.05 and Z=-7.922,P<0.05).CKD3 had higher Lp(a)level than CKD2(Z=-2.069,P<0.05 ),while there were no significant differences between each of the other two groups.CKD4 -5 [(325 .0 ± 194.7)mg/L]also had higher Lp(a)level than CKD1 -3 [(182.0 ±110.5)mg/L](Z=-4.439,P<0.05). Conclusion Patients with CKD always have high level of serum Lp(a),which have been slowly increased since CKD1 ,meanwhile the level of Lp(a)may have a certain correlation with the stage of CKD development,since Lp(a) is an important promoting factor in the progress of CKD.

Objective To explore the clinical phenotype, treatment and prognosis of immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome, and to improve pediatricians' knowledge of this disease. Methods Clinical data of a case of IPEX with congenital ichthyosiform skin lesions were retrospectively analyzed, and related literatures China were reviewed. Results The 2-month-11-day old boy came to our hospital due to ichthyosiform skin lesions accompanied by blood oozing in the head and feet exudatation, with severe sepsis and gastrointestinal perforation. He was died of multiple organ failure. DNA sequencing of whole-genome exon group showed a hemizygous mutation of c.1150G> A, p.A384> T in FOXP3 gene. His mother was a heterozygous mutation carrier, while his father was normal. Conclusions In addition to typical symptoms including early-onset refractory diarrhea, multiple endocrine disease and growth retardation, IPEX should be considered also in infants with ichthyosiform rash and severe infection. Gene sequencing will help diagnose the disease.

Objective To summarize the clinical characteristics,diagnosis and treatment of chronic mucocutaneous candidiasis(CMC).Methods The case diagnosed as CMC in the Department of Nephrology and Immunology of Shenzhen Children's Hospital in February 24,2014 was analyzed in terms of symptoms,signs,laboratory findings,gene tests and treatment process,and related literature was reviewed.Results The patient was a 14-year-old boy.The patient started to develop recurrent oral candida infection shortly after birth,then candida infection of skins and nails,which could be alleviated by antifungal agents,but easily relapsed.Since 4 years ago,autoimmune reactions such as autoimmune anemia,thrombocytopenia,leucopenia,proteinuria,and hypothyroidism had successively appeared,and cytopenia began to palliate after administering Glucocorticoid and Cyclosporin,but easily relapsed when the dosage was reduced.The genetic test showed the case was of signal transducer and activator of transcription 1 (STAT1) gain-of-function mutation.During the hospitalization,hemophagocytic syndrome (HPS) and stagnation of hematopoietic function successively occurred.The cytopenia did not improve and the patient suffered severe infection in spite of washing red blood cells 13 times and blood palate 3 times via infusion,together with high dosage of Dexamethasone and Cyclosporine.The peripheral blood cells and bone marrow gradually returned to normal after being treated by human granulocyte colony-stimulating factor combined with Dexamethasone and Cyclosporin.Retrieving the database in PubMed database,824 articles were found which were about CMC,and 39 of them were about the STAT1 gain-of-function mutation,including 120 cases.But there was only 1 domestic case in 2012,who was a three-year-old child,manifesting recurrent fungal infection of the skin.Conclusions STAT1 gain-of-function acquired mutation is one of the reasons that can lead to CMC.Autoimmune reactions prominently represented by cytopenia occur in a few patients with CMC.It should be alert on those who are with CMC and simultaneously with autoimmtne reaction of blood system.And gene tests facilitate the early diagnosis.

Aim To study the influence of rosuvastatin on toll-like receptor 4 ( TLR4 ) , and its downstream nu-clear transcription factor NF-kappa B p65, IκBα, in-flammation factors TNF alpha in rats with myocardial hypertrophy induced by pressure overload .Methods Myocardial hypertrophy was induced by abdominal aor-tic constriction ( AAC ) .Male Wistar rats were divided into 3 groups(n=10):① sham-operated rats(S);②AAC rats(M);③AAC+rosuvastatin(10 mg· kg -1· d-1 ) rats.From 1 week pre-opertion to 4 weeks post-operation, three groups were performed by gavage ad-ministration with equal volume of rosuvastatin and vehi-cle.The rats underwent cardiac color Doppler exami-nation.The level of ANP,TLR4, NF-κB p65,IκBα, TNF-αmRNA and protein expression in myocardium were detected by real-time PCR, Western blot, immu-nohistochemical and ELISA respectively .Results The sizes of heart and cardiomyocytes and the expression of ANP mRNA and TLR4 signaling molecules were signif-icantly increased in group M , which could be blocked by rosuvastatin .TLR4 protein was positively related to ANP, NF-κB p65 and TNF-αrespectively .Conclusion Rosuvastatin prevents cardiac hypertrophy induced by pressure overload , which is associated with its inhi-bition of TLR4, NF-κB and TNF-αin myocardium ex-pression .