Effects of carbamazepine on BDNF expression in trigeminal ganglia and serum in rats with trigeminal neuralgia / 中南大学学报(医学版)

Objective:Trigeminal neuralgia(TN)is a severe chronic neuropathic pain that mainly affects the distribution area of the trigeminal nerve with limited treating efficacy.There are numerous treatments for TN,but currently the main clinical approach is to suppress pain by carbamazepine(CBZ).Brain-derived neurotrophic factor(BDNF)is closely related to chronic pain.This study aims to determine the effects of CBZ treatment on BDNF expression in both the trigeminal ganglion(TG)and serum of TN via a chronic constriction injury of the infraorbital nerve(ION-CCI)rat model. Methods:The ION-CCI models were established in male Sprague-Dawley rats and were randomly divided into a sham group,a TN group,a TN+low-dose CBZ treatment group(TN+20 mg/kg CBZ group),a TN+medium-dose CBZ treatment group(TN+40 mg/kg CBZ group),and a TN+high-dose CBZ treatment group(TN+80 mg/kg CBZ group).The mechanical pain threshold in each group of rats was measured regularly before and after surgery.The expressions of BDNF and tyrosine kinase receptor B(TrkB)mRNA in TGs of rats in different groups were determined by real-time PCR,and the expression of BDNF protein on neurons in TGs was observed by immunofluorescence.Western Blotting was used to detect the protein expression of BDNF,TrkB,extracellular regulated protein kinases(ERK),and phospho-extracellular regulated protein kinases(p-ERK)in TGs of rats in different groups.The expression of BDNF in the serum of rats in different groups was detected by enzyme-linked immunosorbent assay(ELISA). Results:The results of mechanical pain sensitivity showed that there was no significant difference in the mechanical pain threshold in the right facial sensory area of the experimental rats in each group before surgery(all P>0.05).From the 3rd day after operation,the mechanical pain threshold of rats in the TN group was significantly lower than that in the sham group(all P<0.01),and the mechanical pain threshold of rats in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 CBZ mg/kg group was higher than that in the TN group(all P<0.05).The BDNF and TrkB mRNA and protein expressions in TGs of rats in the TN group were higher than those in the sham group(all P<0.05),and those in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were lower than the TN group(all P<0.05).The p-ERK levels in TG of rats in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were significantly decreased compared with the TN group(all P<0.05).The BDNF and neuron-specific nuclear protein(NeuN)were mainly co-expressed in neuron of TGs in the TN group and they were significantly higher than those in the sham group(all P<0.05).The co-labeled expressions of BDNF and NeuN in TGs of the TN+ 80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were lower than those in the TN group(all P<0.05).The results of ELISA showed that the level of BDNF in the serum of the TN group was significantly higher than that in the sham group(P<0.05).The levels of BDNF in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were lower than those in the TN group(all P<0.05).Spearman correlation analysis showed that the BDNF level in serum was negatively correlated with mechanical pain threshold(r=-0.650,P<0.01). Conclusion:CBZ treatment can inhibit the expression of BDNF and TrkB in the TGs of TN rats,reduce the level of BDNF in serum of TN rats and the phosphorylation of ERK signaling pathway,so as to inhibit TN.The serum level of BDNF can be considered as an indicator for the diagnosis and prognosis of TN.

Wenxiao Powder Alleviates Depression by Promoting Neurogenesis via BDNF/TrkB/ERK/CREB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo decipher the mechanism of Wenxiao powder in alleviating corticosterone-induced depression-like behaviors in mice. MethodMale ICR mice were randomized into normal， model， paroxetine （20 mg·kg-1）， and low- and high-dose （3.27， 6.54 g·kg-1， respectively） Wenxiao powder groups. The mice in normal and model groups received equal volume of saline. Other groups except the normal group were injected with corticosterone subcutaneously 0.5 h after gavage to induce depression. Mice were tested for depression-like behaviors after drug administration. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the corticosterone content in the serum. Nissl staining was performed to observe the damage of hippocampal neurons. Immunofluorescence staining was employed to observe the expression of double cortin （DCX） in the dentate gyrus （DG） of the hippocampus. Western blot was employed to determine the expression of proteins in the brain-derived neurotrophic factor （BDNF）/tyrosine kinase receptor B （TrkB）/extracellular signal-regulated kinase （ERK）/cAMP-response element-binding protein （CREB） pathway in the hippocampus. ResultCompared with the normal group， the model group showed decreased sucrose preference rate， increased immobility time in the tail suspension test （P<0.01）， and reduced residence time in the central area of the open field and the total movement distance （P<0.05， P<0.01）. In addition， the modeling elevated the corticosterone level in the serum （P<0.01）， decreased the volume and intensified the nuclear staining of hippocampal neurons in the DG area， reduced the expression of DCX in the DG area， and down-regulated the protein levels of BDNF， phosphorylated （p）-TrkB， p-ERK， and p-CREB in the hippocampus （P<0.05， P<0.01）. Compared with the model group， low-dose Wenxiao powder improved the mouse behavivors in the sucrose preference， open field， and tail suspension tests （P<0.05， P<0.01）， and high-dose Wenxiao powder improved the behaviors in the sucrose preference and open field tests （P<0.05， P<0.01）. In addition， Wenxiao powder lowered the serum corticosterone level （P<0.01） and recovered the structure and morphology of neurons with obvious nuclei and presence of Nissl bodies in the DG area of the hippocampus. Moreover， Wenxiao powder at both doses promoted the expression of DCX in the DG area， and high-dose Wenxiao powder up-regulated the protein levels of BDNF， p-TrkB， p-ERK， and p-CREB in the hippocampus （P<0.05， P<0.01）. ConclusionWenxiao powder can alleviate corticosterone-induced depression-like behaviors and promote neurogenesis in mice possibly by activating the BDNF/TrkB/ERK/CREB signaling pathway.

Effect of moxibustion on immune function homeostasis in rats with diarrhea irritable bowel syndrome based on SCF/c-kit signaling pathway / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion on the stem cell factor (SCF)/tyrosine kinase receptor (c-kit) signaling pathway and immune function in rats with diarrhea irritable bowel syndrome (IBS-D), and to explore the mechanism of moxibustion for IBS-D.@*METHODS@#Among 52 young rats born from 6 healthy pregnant SPF rats, 12 rats were randomly selected into the normal group, and the remaining 40 rats were treated with the three-factor combination method of maternal separation, acetic acid enema and chronic restraint stress to establish the IBS-D rat model. Thirty-six rats with successful IBS-D model were randomly divided into a model group, a moxibustion group, and a medication group, 12 rats in each group. The rats in the moxibustion group were treated with suspension moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37); the rats in the medication group were treated with intragastric administration of rifaximin suspension (150 mg/kg). All the treatments were given once a day for 7 consecutive days. The body mass, loose stool rate (LSR), the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were measured before acetic acid enema (35 days old), after modeling (45 days old), and after intervention (53 days old). After intervention (53 days old), HE staining was used to observe the morphology of colon tissue, and spleen and thymus coefficients were measured; ELISA method was used to detect serum inflammatory factors (tumor necrosis factor a [TNF-a], interleukin [IL]-10, IL-8), T-lymphocyte subsets (CD+4, CD+8, CD+45), value of CD+4/CD+8 and immune globulin (IgA, IgG, IgM); real-time PCR method and Western blot method was used to detect the expression of SCF, c-kit mRNA and protein in colon tissue; immunofluorescence staining method were used to detect positive expression of SCF and c-kit.@*RESULTS@#After intervention, compared with the normal group, in the model group, the body mass and the minimum volume threshold when AWR scored 3 were decreased (P<0.01), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD+4, CD+45, CD+4/CD+8, IgA, IgG, IgM were increased (P<0.01), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were decreased (P<0.01), and the positive expression of SCF and c-kit was decreased (P<0.01). Compared with the model group, in the moxibustion group and the medication group, the body mass and the minimum volume threshold when AWR scored 3 were increased (P<0.01, P<0.05), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD+4, CD+8, CD+45, CD+4/CD+8, IgA, IgG, IgM were decreased (P<0.01, P<0.05), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were increased (P<0.01), and the positive expression of SCF and c-kit was increased (P<0.01). Compared with the medication group, in the moxibustion group, the level of serum CD+4 was decreased (P<0.05), the value of CD+4/CD+8 was increased (P<0.01), and there was no significant difference in other indexes (P>0.05). The expression of SCF and c-kit mRNA was positively correlated with the minimum volume threshold when AWR scored 3 and IL-10 (P<0.01), and negatively correlated with remaining indexes (P<0.01, P<0.05).@*CONCLUSION@#Moxibustion could reduce visceral hypersensitivity, improve symptoms of abdominal pain and diarrhea in IBS-D rats, and its mechanism may be related to up-regulation of the expression of SCF/c-kit signaling pathway and improvement of IBS-D immune function.

Effect of dexmedetomidine on hippocampal neuron apoptosis and the brain-derived neurotrophic factor/tyrosine kinase receptor B signaling pathway in epileptic rats / 中风与神经疾病杂志

@#Objective To investigate the effect of dexmedetomidine on hippocampal neuron apoptosis and the brain­derived neurotrophic factor（BDNF）/tyrosine kinase receptor B（TrκB） signaling pathway in epileptic rats. Methods A rat model of epilepsy was established by intraperitoneal injection of lithium chloride，atropine sulfate，and pilocarpine hydrochloride. After successful modeling，36 rats were randomly divided into model group，dexmedetomidine（50 μg/kg） group，and dexmedetomidine（50 μg/kg）+LM22B-10（a BDNF/TrκB pathway activator，5 mg/kg） group，with 12 rats in each group，and another 12 rats were selected as control group. After modeling ended，the rats in each group were given corresponding drug intervention once a day for 1 week. At 24 hours after the end of administration，seizure frequency and duration were recorded for each group；TUNEL staining was used to observe hippocampal neuron apoptosis；immunofluorescent staining was used to observe β­tubulin Ⅲ（Tuj1）­positive cells in brain tissue of the hippocampal CA1 region；ELISA was used to measure the levels of tumor necrosis factor­α（TNF­α） and interleukin­6（IL­6） in brain tissue of the hippocampal CA1 region；Western blot was used to measure the protein expression levels of BDNF，TrκB，B­cell lymphoma­2 associated X protein（Bax），and B cell lymphoma­2（Bcl­2） in brain tissue of the hippocampal CA1 region. Results Compared with the control group，the model group had significant increases in seizure frequency and duration，the number of apoptotic hippocampal neurons，the levels of TNF­α and IL­6 in the hippocampal CA1 region，and the protein expression levels of BDNF，TrκB，and Bax in the hippocampal CA1 region（P<0.05） and significant reductions in the number of Tuj1­positive cells and the protein expression level of Bcl-2 in the hippocampal CA1 region（P<0.05）. Compared with the model group，the dexmedetomidine group had significant reductions in seizure frequency and duration，the number of apoptotic hippocampal neurons，the levels of TNF-α and IL­6 in the hippocampal CA1 region，and the protein expression levels of BDNF，TrκB，and Bax in the hippocampal CA1 region（P<0.05） and significant increases in the number of Tuj1­positive cells and the protein expression level of Bcl­2 in the hippocampal CA1 region（P<0.05）. LM22B­10 could partially reverse the role of dexmedetomidine in improving various indicators of epileptic rats（P<0.05）. Conclusion Dexmedetomidine can reduce seizure in epileptic rats，inhibit the apoptosis of hippocampal neurons，and alleviate inflammatory response in rats，possibly by inhibiting the activation of the BDNF/TkB signal pathway.

Effect of electroacupuncture at "Zusanli" (ST 36) on duodenal mast cells, NGF and NTRK1 in rats with functional dyspepsia / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) on duodenal mast cells, nerve growth factor (NGF) and neurotrophic tyrosine kinase receptor type 1 (NTRK1), and to explore the mechanism of electroacupuncture at Zusanli (ST 36) on functional dyspepsia (FD).@*METHODS@#Sixty SPF-grade 10-day-old SD rats were randomly divided into a normal group, a model group, a ketotifen group and an EA group, 15 rats in each group. The FD model was prepared by iodoacetamide combined with rat tail clamping method in the model group, the ketotifen group and the EA group. The rats in the ketotifen group were injected intraperitoneally with ketotifen (1 mg•kg-1•d-1) for 7 days; the rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), with disperse-dense wave, frequency of 2 Hz/50 Hz and intensity of 0.5 mA, 20 min each time, once a day for 14 days. The gastric emptying rate and small intestinal propulsion rate in each group were observed; the morphology of duodenal mucosa was observed by HE staining; the toluidine blue staining was used to observe the number and degranulation of mast cells in duodenal mucosa; the protein and mRNA expressions of NGF, NTRK1 in duodenum were detected by Western blot and real-time PCR; the level of interleukin-1β (IL-1β) in duodenum was measured by ELISA.@*RESULTS@#Compared with the normal group, the gastric emptying rate and small intestinal propulsion rate in the model group were decreased (P<0.01); compared with the model group, the gastric emptying rate and small intestinal propulsion rate in the ketotifen group and the EA group were increased (P<0.01); the small intestinal propulsion rate in the EA group was higher than that in the ketotifen group (P<0.01). In the model group, local defects in duodenal mucosa were observed with a small amount of inflammatory cell infiltration; no obvious abnormality was found in duodenal mucosa of the other groups. Compared with the normal group, the mast cells of duodenal mucosa in the model group were increased significantly with significant degranulation; compared with the model group, the mast cells of duodenal mucosa in the ketotifen group and the EA group were decreased significantly, and the degranulation was not obvious. Compared with the normal group, the protein and mRNA expressions of NGF, NTRK1 as well as the level of IL-1β in duodenum in the model group were increased (P<0.01); compared with the model group, the protein and mRNA expressions of NGF, NTRK1 as well as the levels of IL-1β in duodenum in the ketotifen group and the EA group were decreased (P<0.01, P<0.05); compared with the ketotifen group, the mRNA expression of NGF, as well as the protein and mRNA expressions of NTRK1 in duodenum in the EA group were decreased (P<0.05, P<0.01).@*CONCLUSION@#EA at "Zusanli" (ST 36) could inhibit the activation of duodenal mast cells and regulate the expressions of NGF and its receptor to improve the low-grade inflammatory response of duodenum, resulting in treatment effect on FD.

Effects of overexpression of tyrosine kinase receptor 1 on malignant biological behavior of tumor-associated endothelial cells in cervical cancer cells / 安徽医科大学学报

Objective @#To investigate the effect of overexpression of tyrosine kinase receptor 1 (Tie-1) in cervical cancer cells on the malignant biological behavior of tumor-related endothelial cells (TRECs) ．@*Methods @#Immuno- histochemical method was used to detect the expression of Tie-1 in cervical cancer cells ( CCCs) and TRECs of 96 patients with cervical cancer，and to analyze the correlation between the expression of Tie-1 in TRECs and clinico- pathological features and prognosis of patients．HeLa cells overexpressing Tie-1 (Hela-Tie1OE) were constructed， and HeLa-Tie1OE was co-cultured with human umbilical vein endothelial cells ( HUVECs) by Transwell cell co- culture method to obtain cervical cancer TRECs．Western blot was used to analyze Tie-1 protein expression．The migration，invasion and tubulogenesis of TRECs were detected by cell scratch assay，Transwell invasion and migra- tion assay and tubulogenesis assay. @*Results @#The expression of Tie-1 was positively correlated with CCCs and TRECs in 96 cervical cancer patients．The positive expression rate of Tie-1 in TRECs of patients with stage(FIGO) Ⅰ B2-ⅡA，tumor diameter ≥4 cm ，cervical muscle invasion depth ≥ 1 /2 full layer ，adenocarcinoma ，medium and low differentiation cervical cancer was higher than that of patients with Ⅰ A 1-Ⅰ B 1，tumor diameter＜4 cm， cervical muscle invasion depth ＜1 /2 full layer，squamous carcinoma，and high differentiation cervical cancer，re- spectively．The differences were statistically significant (P＜0. 05) ．The expression of Tie-1 in TRECs of cervical cancer patients had no significant correlation with age，lymph node metastasis and lymphatic space invasion．The positive expression of Tie-1 in cervical cancer TRECs was negatively correlated with 5-year progression-free survival time and overall survival time (P＜0. 05) ．The Tie-1 expression of TRECs obtained by co-culture of Hela-Tie1OE and HUVECs was up-regulated ，and the invasion ，migration and tubulogenesis of TRECs cells were enhanced. @*Conclusion @#The high expression of Tie-1 in TRECs of cervical cancer is related to FIGO stage，tumor diameter， degree of differentiation，depth of cervical muscular invasion，type of cervical cancer，and poor prognosis of pa- tients．Overexpression of Tie-1 can promote TRECs invasion，migration and tubulogenesis.

Effect of Sinisan on BDNF/TrkB， 5-HT/5-HT1AR， and HPA axis in Depression Model Rats / 中国实验方剂学杂志

Objective:To observe the effect of Sinisan on the brain-derived neurotrophic factor （BDNF）/tyrosine kinase receptor B （TrKB）， 5-hydroxytryptamine （5-HT）/5-HT1A receptor （5-HT1AR）， and hypothalamus-pituitary-adrenal （HPA） axis in depressed rats， and explore the antidepressant mechanism of Sinisan based on BDNF/TrKB， 5-HT/5-HT1AR， and HPA axis. Method:A total of 120 male Wistar rats were randomly divided into a normal group， a model group， a fluoxetine （0.01 g·kg^-1） group， and low- （1.25 g·kg^-1）， medium- （2.5 g·kg^-1）， and high-dose （5 g·kg^-1） Sinisan groups， with 20 rats in each group. The depression model was induced by isolation combined with chronic unpredictable mild stimulation（CUMS） in rats except for those in the normal group for 21 days. Rats were then treated correspondingly once a day for 21 days by gavage. Those in the normal group and the model group received an equal volume of normal saline. During the intervention， the model rats were stimulated continuously. The depressive state of CUMS model rats was evaluated by sucrose preference test and open field test. Enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of corticotropin-releasing hormone （CRH）， adrenocorticotropic hormone （ACTH）， and corticosterone （CORT） in the plasma and BDNF and 5-HT levels in the hippocampal homogenate. The mRNA expression of hippocampal TrKB， 5-HT1AR， glucocorticoid receptor （GR）， and mineralocorticoid receptor （MR） was detected by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）. The protein expression of hippocampal TrKB， 5-HT1AR， GR， and MR was detected by Western blot. The histomorphological changes of the hippocampus were observed by hematoxylin-eosin （HE） staining. Result:Compared with the normal group， the model group showed decreased sucrose preference rate （<italic>P</italic><0.01）， reduced horizontal and vertical scores in the open field test （<italic>P</italic><0.01）， increased plasma content of CRH， ACTH， and CORT （<italic>P</italic><0.01）， declining content of BDNF and 5-HT in the hippocampus （<italic>P</italic><0.01）， dwindled mRNA and protein expression levels of TrKB， 5-HT1AR， and GR （<italic>P</italic><0.01）， elevated mRNA and protein expression of MR （<italic>P</italic><0.01）， and damaged hippocampal neurons revealed by HE staining. Compared with the model group， the groups with drug intervention showed increased sucrose preference rate （<italic>P</italic><0.01） and horizontal and vertical scores in the open field test （<italic>P</italic><0.05， <italic>P</italic><0.01）， decreased content of plasma CRH， ACTH， and CORT （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated content of hippocampal BDNF and 5-HT （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated mRNA and protein expression levels of hippocampal TrKB， 5-HT1AR， and GR （<italic>P</italic><0.05， <italic>P</italic><0.01）， reduced mRNA and protein expression levels of hippocampal MR （<italic>P</italic><0.05， <italic>P</italic><0.01）， and recovered hippocampal neurons as revealed by HE staining. Conclusion:Sinisan can exert a significant antidepressant effect by increasing hippocampal BDNF and 5-HT content， up-regulating TrKB， 5-HT1AR， and GR mRNA and protein expression， down-regulating MR mRNA and protein expression， inhibiting HPA axis hypertrophy， and enhancing the regeneration and repair of hippocampal neurons.

Importancia clínica y diagnóstica de la relación receptor de tirosin-quinasa tipo 1 en su forma soluble y el factor de crecimiento placentario / Clinical and diagnostic importance of the type 1 receptor tyrosine-kinase ratio in its soluble form and placental growth factor / Importância clínica e diagnóstica da relação receptor de tirosina-quinase tipo 1 em sua forma solúvel e o fator de crescimento placentário

El objetivo del trabajo fue evaluar la utilidad clínica de la relación factor de crecimiento placentario/receptor de tirosin-quinasa tipo 1 soluble (sFlt-1/ PlGF) para el diagnóstico de preeclampsia (PE) en embarazadas de alto riesgo y con diagnóstico clínico de PE en un centro de salud de Córdoba, Argentina. Se procesaron 135 muestras de embarazadas: 39 con diagnóstico clínico de PE (Grupo I), 72 con riesgo de PE (Grupo II), y 24 de grupo control (Grupo III). Se utilizó una técnica automatizada de electroquimioluminiscencia (Roche). Valores <38 se consideraron sin riesgo de PE, entre 38 y 85 (< semana 34) o 38 y 110 (> semana 34) con riesgo moderado o alto riesgo dentro de las 4 semanas posteriores a la realización de dichos marcadores y >85 en embarazadas con síntomas de aparición temprana o >110 en embarazadas con síntomas de aparición tardía, PE confirmada. En el Grupo I, 33 muestras dieron relación >38 y 6 fueron menores. De 72 muestras del Grupo II 69 dieron <38 y 3 >38. Todas las muestras del Grupo III dieron relación <38. La razón de verosimilitud positiva (LR+) fue de 20,31 y la razón negativa (LR-) fue de 0,16. La relación fue >38 en la mayoría de las embarazadas con diagnóstico de PE. La determinación es útil en aquellas mujeres embarazadas que son de alto riesgo, ya sea porque tienen hipertensión o proteinuria o algún antecedente previo, en las cuales puede ser fundamental para decidir el correcto diagnóstico.

The objective of this work was to evaluate the clinical usefulness of the relation placental growth factor/soluble tyrosine-kinase type 1 receptor (sFlt-1/PlGF) for the diagnosis of PE (preeclampsia) in pregnant women at high risk and with clinical diagnosis of PE in a health center of Córdoba, Argentina. A total of 135 samples of pregnant women were processed: 39 with clinical diagnosis of PE (Group I), 72 with risk of PE (Group II), and 24 of control group (Group III). An automated electrochemiluminescence technique (Roche) was used. Ratio sFlt-1/PlGF <38 was considered without risk of PE. Between 38 and 85 (< week 34) or 38 and 110 (> week 34), with moderate risk or high risk within 4 weeks after performing these markers. To confirm diagnosis, relationships >85 in pregnant women were considered with symptoms of early onset and >110 in pregnant women with symptoms of late onset. In Group I, 33 samples reported >38 and 6 were lower. Of 72 samples from Group II, 69 gave <38 and 3, >38. All samples from Group III gave a ratio <38. The positive likelihood ratio (LR+) was 20.31 and the negative likelihood ratio (LR-), 0.16. The ratio was >38 in the majority of women already diagnosed as PE. The test is useful in those pregnant women who are at high risk of PE, either because they have hypertension or proteinuria or a previous history. In those cases it can be fundamental to decide the correct diagnosis.

O objetivo do estudo é avaliar a utilidade clínica da relação fator de crescimento placentário/receptor de tirosina-quinase tipo 1 solúvel (sFlt-1/PIGF) para o diagnóstico de pré-eclâmpsia (PE) em grávidas de alto risco e com diagnóstico clínico de PE em um centro de saúde de Córdoba, Argentina. Foram processadas 135 amostras de gestantes, sendo 39 com diagnóstico clínico de PE (Grupo I), 72 com risco de PE (Grupo II) e 24 de grupo controle (Grupo III). Foi utilizada uma técnica automatizada de eletroquimioluminescência (Roche). Valores <38 foram considerados sem risco de PE, entre 38 e 85 (

Anti-depressant Mechanism of Qing' ewan in CUMS Rats Based on Estrogen Receptor Pathway / 中国实验方剂学杂志

Objective::To study the anti-depressive effect of Qing' ewan in treating chronic unpredictable mild stress (CUMS) in rats, and the regulatory effect on estrogen receptor and estrogen receptor-related signaling pathways, in order to explore its anti-depressive mechanism. Method::The CUMS model was established. The experiment was divided into normal control group, model group, escitalopram oxalate group (positive control) and Qing' ewan groups (1.71, 5.13, 15.39 g·kg-1). After 4 weeks of modeling, rats were treated with corresponding drugs for 2 weeks. Behavioral evaluation [sucrose preference test (SPT), forced swimming test (FST), open field test (OFT)] was conducted to assess if the CUMS model was successful. Western blot was used to analyze the protein expression levels of estrogen receptor α (ERα), estrogen receptor β (ERβ), brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB). Result::Compared with the normal group, the sucrose consumption rate and the score of OFT in the model group decreased(P<0.05, P<0.01), the immobility time of FST prolonged significantly(P<0.01), and the protein expression levels of ERα, ERβ, BDNF and TrkB decreased(P<0.05, P<0.01). Compared with the model group, the behavioral performance of the treated group was improved, the sucrose consumption rate and the score of OFT increased(P<0.05, P<0.01), and the immobility time decreased(P<0.05). The protein expressions of ERα, ERβ, BDNF and TrkB in the treated group were significantly up-regulated(P<0.05, P<0.01), especially the middle-dose Qing' ewan group (5.13 g·kg-1). Conclusion::Qing' ewan can improve depression-like behavior in CUMS rats. Its mechanism may be related to the neuroprotective effect by up-regulating the expressions of ERα and ERβ and activating estrogen receptor-mediated ERβ/BDNF/TrkB pathways.

Neuroprotective Effect by Wutoutang in Spinal Nerve Ligation Mice via BDNF/TrkB Signaling Pathway / 中国实验方剂学杂志

Objective::To investigate the neuroprotective effect and mechanism by Wutoutang (WTT) in the spinal nerve ligation (SNL) mice by neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrkB) signaling BDNF/tyrosine kinase receptor B (TrkB). Method::The 40 mice were randomly divided into Sham group, SNL group, WTT group(126 mg·kg-1), ANA-12+ WTT group.The L5 spinal nerve ligation model mice were established in mice, After that, WTT was administrated from the first day to the 10th day, then the consecutive 7-day hippocampal injection of ANA-12(0.05 nmol·L-1)were lasted for 7 days.The levels of brain-derived BDNF, cAMP-response element binding protein(CREB), and protein kinase B(Akt)and the change of hippocampal glutamatergic and GABAergic neurons in mice were detected by tissue immunofluorescence.E14 pregnant ICR mice were sacrificed and the hippocampus were dissected which were divided into control group, glycine group, tumo necrosis factor(TNF)-α(5 mg·L-1)+ glycine group, TNF-α+ WTT(5 mg·L-1)+ glycine group, TNF-α+ WTT+ glycine+ BDNF-siRNA group, TNF-α+ WTT+ glycine+ Akt-siRNA group, TNF-α+ WTT+ glycine+ CREB-siRNA group, the primary and secondary dendrrictes, in which the arrowheads indicate the expression od postsynapti desity protein 95(PSD95) in the shafts and arrows were tested by cellular immunofluorescence.The neurons were divided into control group, glycine group, ANA-12 group(0.5 mmol·L-1), ANA-12+ glycine group, ANA-12+ WTT group, ANA-12+ WTT+ glycine group, the morphology of hippocampal neurons were tested by cellular immunofluorescence. Result::Compared with Sham group, BDNF, Akt and CREB positive cell of SNL group decreased significantly(P<0.01), the hippocampal glutamatergic and GABAergic neurons out of balance(P<0.01). Compared with SNL group, the BDNF, Akt and CREB positive cell of WTT group increased significantly(P<0.01), Glutamine-aminobutyric acid neurons regein balance(P<0.01). Compared with WTT group, BDNF and CREB positive cell of ANA-12+ WTT group decreased significantly(P<0.05), Glutamine-aminobutyric acid neurons was disorderedd(P<0.05). Comparaed with control group, the level of PSD95 of glycine group were increase significantly(P<0.01). The number of dendritic spine density apically and basally of glycine group were increase significantly(P<0.01), but the primary and secondary dendrites of ANA-12 group, ANA-12+ glycine group, ANA-12+ WTT group, ANA-12+ WTT+ glycine group were not change.Comparaed with glycine group, the level of PSD95 of TNF-α+ glycine group were decreased significantly(P<0.01). Comparaed with TNF-α+ glycine group, the level of PSD95 of TNF-α+ WTT+ glycine group were increase significantly(P<0.01). Comparaed with TNF-α+ WTT+ glycine group, the level of PSD95 of TNF-α+ WTT+ glycine+ BDNF-siRNA group, TNF-α+ WTT+ glycine+ Akt-siRNA group, TNF-α+ WTT+ glycine+ CREB-siRNA group were decreased significantly(P<0.01). Conclusion::In vivo and in vitro studies have shown that the WTT mediated remission of the primary hippocampal glutamatergic neurons were dependent on the BDNF/TrkB pathway.

Research progress on the role of Ang/Tie axis in angiogenesis and metastasis / 药学学报

The tumor contains abundant new vessels, which are unevenly distributed, irregular, and branch-disordered. Angiopoietin (Ang) and tyrosine kinase receptor Tie mediate stable maturation of angiogenesis. Ang1 mainly plays a role in promoting vascular stabilization, and Ang2 is highly expressed in vessels, which makes the structure and function of vessels abnormal. Leaked vessels provide opportunities for invasion and metastasis of circulating tumor cells. Targeting the Ang/Tie axis to correct the abnormal state of vessels and promote its normalization, combined with chemotherapy drugs or immunotherapy, play a synergistic effect against tumors. This article summarizes the role of Ang/Tie axis in tumor angiogenesis and metastasis, and it aims to provide new ideas and strategies for clinical treatment of tumors.

New Research of Nerve Growth Factor on Fracture Healing / 中国医学科学院学报

Fracture healing has long been a major concern for orthopedists.Currently,about 10% of fracture patients still have poor fracture healing or bone nonunion.In recent years,research has found that nerve growth factor(NGF)can promote fracture healing.This article reviews the mechanism and research progress of NGF in promoting fracture healing.NGF can promote vascular regeneration and nerve growth at callus and plays an important role in the proliferation and migration of bone cells.New animal experiments and clinical trials have confirmed the role of NGF in promoting fracture healing and further revealed its possible mechanism of action.Further research on NGF can provide new ideas for promoting fracture healing,especially for treating nonunion.

Effect of Tamibarotene on the proliferation and Tyrosine kinase receptor A,N-myc expressions of neuroblastoma SH-SY5Y cells / 中华实用儿科临床杂志

Objective To study the effect of Tamibarotene on the SH-SY5Y cell proliferation inhibition ability and the mRNA and protein expressions of tyrosine kinase receptor a (TrkA) and N-myc (MYCN) in order to provide some experimental bases for the treatment of neuroblastoma.Methods The SH-SY5Y cells were treated with different concentrations of Am80 (0,10,20,40,80,160 μmol/L) for 48 h,then Cell Counting Kit-8 (CCK-8) was used to test the cell proliferation.Reverse transcription PCR(RT-PCR) and Western blot were used to test the mRNA and protein expressions of TrkA and MYCN at 48 hours.Results When the concentration was 10 μmol/L,Am80 had no significant inhibitory effect on SH-SY5Y cells [(3.51 ± 1.68)％,inhibition ratio ＜ 5 ％];but when the concentration was 20 μmol/L,it showed weak inhibition [(9.60 ± 1.97) ％,inhibition ratio ＜ 10％].The inhibition rate of SH-SY5Y cell proliferation[(57.43 ± 4.95)％] was significantly enhanced at Am80 with a concentration of 80 μmol/L.The concentrations of Am80 could effectively inhibit SH-SY5Y cell proliferation in a dose-dependent manner(P ＜0.05).The expression of TrkA increased with the increase of Am80 concentration.Am80 significantly decreased the expression of MYCN in SH-SY5Y cells(10 μmol/L:0.65 ±0.05 vs.20 μmol/L:0.36 ±0.06),and the difference was statistically significant(P ＜ 0.05).Conclusions It is suggested that Am80 can effectively inhibit SH-SY5Y cell proliferation in a concentration-dependent manner.The underlying mechanism involves increasing the expression of TrkA by down-regulation of MYCN.

Discovery of a series of dimethoxybenzene FGFR inhibitors with 5pyrrolo2,3-pyrazine scaffold: structure-activity relationship, crystal structural characterization and study

Genomic alterations are commonly found in the signaling pathways of fibroblast growth factor receptors (FGFRs). Although there is no selective FGFR inhibitors in market, several promising inhibitors have been investigated in clinical trials, and showed encouraging efficacies in patients. By designing a hybrid between the FGFR-selectivity-enhancing motif dimethoxybenzene group and our previously identified novel scaffold, we discovered a new series of potent FGFR inhibitors, with the best one showing sub-nanomolar enzymatic activity. After several round of optimization and with the solved crystal structure, detailed structure-activity relationship was elaborated. Together with metabolic stability tests and pharmacokinetic profiling, a representative compound () was selected and tested in xenograft mouse model, and the result demonstrated that inhibitor was effective against tumors with FGFR genetic alterations, exhibiting potential for further development.

Effects of Enriched Environment on Apoptosis and Brain-derived Neurotrophic Factor /Tyrosine Kinase Receptor B Signaling Pathway in Hippocampal Neurons of Neonatal Rats with Hypoxic-ischemic Brain Damage / 中国康复理论与实践

Objective:To investigate the effects of enriched environment on hippocampal neuronal apoptosis and brain-derived neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrkB) signaling pathway in neonatal rats with hypoxic-ischemic brain damage. Methods:Forty-eight newborn Wistar rats aged seven days were randomly divided into sham operation group, model group and enriched environment group, each group was divided in to 14 days group and 28 days group, with eight in each subgroup. The model was established with the Rice method. The sham operation group and the model group did not receive any intervention, and the enriched environment group received enriched environment stimulation 24 hours after modeling. Fourteen days and 28 days after modeling, the levels of neuronal apoptosis in hippocampus were detected by TUNEL and double immunofluorescence staining; BDNF and TrkB proteins in hippocampus were detected by immunohistochemical staining. Results:Fourteen days and 28 days after modeling, the numbers of TUNEL positive cells, double immunofluorescence positive cells, BDNF and TrkB positive cells were significantly more in the model group than in the sham operation group (t > 27.214, P < 0.001), while the numbers of TUNEL positive cells, double immunofluorescence positive cells were significantly less in the enriched environment group than in the model group (t > 12.687, P < 0.001); and the number of BDNF and TrkB positive cells were significantly more in the enriched environment group than in the model group 28 days after modeling (t > 137.998, P < 0.001). Conclusion:Enriched environmental stimulation could reduce the apoptosis of hippocampal neurons, and up-regulate the expression of BDNF and TrkB proteins in the neonatal rats with hypoxic-ischemic brain damage.

Renal developmental retardation and the underlying mechanism induced by prenatal caffeine exposure in female fetal rats / 中国药理学通报

Aim To explore the effects of prenatal caf-feine exposure (PCE) on fetal renal growth retardation and corticosterone on the gene expression of metanephric mesenchyme stem cells.Methods Pregnant Wistar rats were administered with caffeine (30,120 mg ·kg-1) from gestational day 9 to 20.Female fetal kidney samples were collected for morphological observation and gene expression examination.The metanephric mesenchyme stem cells were harvested for cell culture,and renal related genes were detected after the treatment of corticosterone with different concentrations (250,500,1 000 μg · L-1) for 24 hours.Results Compared with the control group,the fetal kidneys in the PCE group displayed an enlarged Bowman's space and a shrunken glomerular tuft,accompanied with the repression of the gene expression of glial-cell-line-derived neurotrophic factor/tyrosine kinase receptor (GDNF/c-Ret) signaling pathway.The GDNF/c-Ret signaling pathway and angiotensin Ⅱ receptor type 1 (AT1R)/AT2R expression of metanephric mesenchyme stem cells also decreased in corticosterone groups.Conclusions PCE may induce dysplasia of female fetal kidneys.The potential mechanism is related to the repression of the gene expression of AT1R/AT2R and GDNF/c-Ret signaling pathway by PCE mediated by corticosterone in utero.

Low frequency repetitive nerve stimulation in anti-muscle specific tyrosine kinase receptor antibody positive myasthenia gravis / 中华神经科杂志

Objective To describe the repetitive nerve stimulation ( RNS) in anti-muscle specific tyrosine kinase (anti-MuSK) receptor antibody positive myasthenia gravis (MG), and compare with anti-acetylcholine receptor ( AChR ) positive myasthenia gravis , to figure out characteristics of anti-MuSK receptor MG.Methods We analyzed clinical and RNS data of nine anti-MuSK receptor MG and 19 age-and sex-matched anti-AChR MG.RNS was performed to the abductor digiti minimi , orbicularis oculi or musculus frontalis and trapezius .Results In anti-MuSK receptor MG , abnormal RNS in facial nerve was seen in 6/9 and in trapezius was 5/9, in limbs was 0.In anti-AChR MG, abnormal RNS in facial nerve was seen in 13/19, in trapezius was 18/19 and in limbs was 7/19.Abnormal in any of three parts was 8/9 and 19/19 in anti-MuSK receptor MG and anti-AChR MG, respectively.The RNS decrementing was more obvious in facial nerve in anti-Musk receptor MG than in anti-AChR MG.Negative prostigmin test was independently associated with anti-MuSK receptor MG (OR=4.25,95% CI 2.19 -15.25, P=0.015). Conclusions Abnormal RNS in any of three parts is more pronounced in anti-AChR MG compared with anti-MuSK receptor MG.RNS decrementing in facial nerve is more obvious in anti-AChR MG.Negative prostigmin test can aid in early suspicion in anti-MuSK receptor MG.

Low frequency repetitive nerve stimulation in anti-muscle specific tyrosine kinase receptor antibody positive myasthenia gravis / 中华神经科杂志

Objective To describe the repetitive nerve stimulation ( RNS) in anti-muscle specific tyrosine kinase (anti-MuSK) receptor antibody positive myasthenia gravis (MG), and compare with anti-acetylcholine receptor ( AChR ) positive myasthenia gravis , to figure out characteristics of anti-MuSK receptor MG.Methods We analyzed clinical and RNS data of nine anti-MuSK receptor MG and 19 age-and sex-matched anti-AChR MG.RNS was performed to the abductor digiti minimi , orbicularis oculi or musculus frontalis and trapezius .Results In anti-MuSK receptor MG , abnormal RNS in facial nerve was seen in 6/9 and in trapezius was 5/9, in limbs was 0.In anti-AChR MG, abnormal RNS in facial nerve was seen in 13/19, in trapezius was 18/19 and in limbs was 7/19.Abnormal in any of three parts was 8/9 and 19/19 in anti-MuSK receptor MG and anti-AChR MG, respectively.The RNS decrementing was more obvious in facial nerve in anti-Musk receptor MG than in anti-AChR MG.Negative prostigmin test was independently associated with anti-MuSK receptor MG (OR=4.25,95% CI 2.19 -15.25, P=0.015). Conclusions Abnormal RNS in any of three parts is more pronounced in anti-AChR MG compared with anti-MuSK receptor MG.RNS decrementing in facial nerve is more obvious in anti-AChR MG.Negative prostigmin test can aid in early suspicion in anti-MuSK receptor MG.

Effect of up-regulation of miR-132 on expressions of angiopoietin-1/endothelium-specific tyrosine kinase receptor 2 in focal cerebral ischemia reperfusion injury of rats / 中华神经医学杂志

Objective To investigate the effect of up-regulation of miR-132 on expressions of angiopoietin-1 (Ang-1)/endothelium-specific tyrosine kinase receptor 2 (Tie2) in focal cerebral ischemia reperfusion injury of rats.Methods Forty adult healthy SD rats were randomly divided into sham-operated group,cerebral ischemia group,miR-132 mimic group and negative control group (n=10).The models of middle cerebral artery occlusion in the later three groups were established by using modified Longa suture method.Rats in the miR-132 mimic group and negative control group were injected miR-132 mimic 15 μg and negative control 15 μg via paracele.Rats in each group were sacrificed 24 h after ischemia,and the brain tissues were collected;the total infarct volumes were calculated by TTC staining.The mRNA expressions ofmiR-132,Ang-1 and Tie2 in ischemic cerebral cortex tissues were detected by real-time fluorescence quantitative PCR.The protein expressions of Ang-1,Tie2,CD31 and vessel endothelial growth factor (VEGF) in ischemic cerebral cortex tissues were detected by Western blotting.Results The total infarct volume in the miR-132 mimic group was (27.92±3.05) mm3,which was significantly smaller than that in the cerebral ischemia group and negative control group ([51.34±2.86] mm3 and [50.46±2.57] mm3,P＜0.05).The relative mRNA expression levels of miR-132,A ng-1,Tie2,and the protein expression levels of Ang-1,Tie2,CD31,VEGF in the ischemic cerebral cortex tissues of the miR-132 mimic group were significantly higher than those in the negative control group,cerebral ischemia group and sham-operated group (P＜0.05);and those in the negative control group and cerebral ischemia group were significantly higher than those in the sham-operated group (P＜0.05).Conclusion Up-regulation ofmiR-132 expression could improve the ischemic states of ischemic stroke in rats,which might be related to Ang-1/Tie2 increased expressions to promote angiogenesis in ischemic brain tissues.

Role of tyrosine kinase receptor RON in tumor growth and metastasis:research progress / 中国药理学与毒理学杂志

Recepteur d′origine Nantais(RON),a tyrosine kinase receptor ,is a growth factor receptor belonging to the proto-oncogene met family and has been proved to display abnormal expres?sion in many types of tumors. The RON receptor is activated by binding to the ligand macrophage stim?ulating protein,overexpression of the receptor,variants of the proto-oncogene and by point mutations of the kinase region. The downstream transduction of RON by mitogen-activated protein kinases and phosphoinositide3-kinase signaling pathways can help regulate the proliferation,apoptosis,migration and invasion of tumor cells. A better understanding of the mechanisms and related signaling pathways of RON activation in tumor progress and development will provide more information for the RON-based target therapy.