Tong xie yao fang relieves irritable bowel syndrome in rats via mechanisms involving regulation of 5-hydroxytryptamine and substance P

AIM: To investigate whether the Chinese medicine Tong Xie Yao Fang(TXYF) improves dysfunction in an irritable bowel syndrome(IBS) rat model. METHODS: Thirty baby rats for IBS modeling were separated from mother rats(1 h per day) from days 8 to 21, and the rectum was expanded by angioplasty from days 8 to 12. Ten normal rats were used as normal controls. We examined the effects of TXYF on defection frequency, colonic transit function and smooth muscle contraction, and the expression of 5-hydroxytryptamine(5-HT) and substance P(SP) in colonic and hypothalamus tissues by Western blot and RT-PCT techniques in both normal rats and IBS model rats with characterized visceral hypersensitivity.in normal rats and 4.5 ± 1.58 in IBS model rats(P < 0.001). However, the defecation frequency was significantly decreased(3.0 ± 1.25 vs 4.5 ± 1.58, P < 0.05), while the time(in seconds) of colon transit function was significantly increased(256.88 ± 20.32 vs 93.36 ± 17.28, P < 0.001) in IBS + TXYF group rats than in IBS group rats. Increased colonic smooth muscle tension and contract frequency in IBS model rats were significantly decreased by administration of TXYF. Exogenous agonist stimulants increased spontaneous activity and elicited contractions of colon smooth muscle in IBS model rats, and all of these actions were significantly reduced by TXYF involving 5-HT and SP down-regulation. CONCLUSION: TXYF can modulate the activity of the enteric nervous system and alter 5-HT and SP activities, which may contribute to the symptoms of IBS.

Changes of 5-hydroxytryptamine and tryptophan hydroxylase expression in the ventral horn of spinal cord

Objective To investigate changes of 5-hydroxytryptamine （5-HT） and its synthesis rate-limiting enzyme tryp-tophan hydroxylase （TPH） in the ventral horn of spinal cord after exercise-induced fatigue, and to further discuss the mecha- nism of exercise-induced central fatigue at spinal level. Methods Sixteen healthy adult Wistar rats were randomly divided into 2 groups： exercise-induced fatigue group and control group. Immunohistochemical staining for 5-HT and TPH in the ventral horn were performed and analysized quantitatively. The mean optic densities of 5-HT and TPH positive fibers or terminals were measured by computerized image analyzer. Results Both 5-HT and TPH positive fibers/terminals decreased in the exercise-induced fatigue group. The immunohistochemical staining was weaker and the mean optic densities decreased obviously in the fatigue group compared with those in the control group （P 〈 0.05）. Conclusion 5-HT and TPH in the ventral horn of spinal cord might be involved in exercise-induced fatigue.

Effect of the pineal gland on 5-hydroxytryptamine and γ-aminobutyric acid secretion in the hippocampus of male rats during the summer and winter

Objective:The present study aimed to investigate the effect of seasonal variation on neurotransmitter release in the hippocampus of normal rats and rats with pineal excision.Methods:Two time points,the summer and winter solstice,which are the longest and shortest days of the year,respectively,were selected.Male Spraguee Dawley rats that underwent a sham operation without pineal excision were included as a control group.The concentrations of 5-hydroxytryptamine(5-HT)andγ-aminobutyric acid(GABA)were determined by radioimmunoassays and enzyme-linked immunosorbent assays,respectively.Results:In the winter,the 5-HT and GABA levels in normal rats exhibited a significant difference compared with those in the operation group(P<.01).A difference was also noted in GABA levels between the normal group and the sham operation group(P<.05).The concentrations of 5-HT and GABA in the hippocampal tissues of the normal group exhibited a seasonal rhythm consisting of elevation during the summer and reduction during the winter(P<.01),while the GABA levels in the sham operation group exhibited a significant difference,with elevation during the summer and reduction during the winter(P<.01).In the operation group,GABA showed the same trend(P<.01).Conclusion:The seasonal rhythm of neurotransmitter secretion by the hippocampus(5-HT and GABA)consisted of elevation during the summer and reduction during the winter.During the winter,the pineal gland exhibited a reverse regulatory effect on the secretion of 5-HT and GABA in the hippocampus,and it exhibited seasonal selectivity with regard to the regulation of 5-HT.

5-Hydroxytryptamine:a potential therapeutic target in amyotrophic lateral sclerosis

Previous studies have indicated that the pathogenesis of amyotrophic lateral sclerosis(ALS) is closely linked to 5-hydroxytryptamine(5-HT).To investigate this further,we administered 5-HT receptor antagonists to SOD1*G93A transgenic(ALS mouse model) and wide-type mice.This involved intraperitoneal injections of either granisetron,piboserod,or ritanserin,which inhibit the 5-HT3,5-HT4,and 5-HT2 receptors,respectively.The transgenic mice were found to have fewer5-HT-positive cells in the spinal cord compared with wide-type mice.We found that the administration of granisetron reduced the body weight of the transgenic mice,while piboserod and ritanserin worsened the motor functioning,as assessed using a hanging wire test.However,none of the 5-HT receptor antagonists affected the disease progression.We analyzed the distribution and/or expression of TAR DNA binding protein 43(TDP-43) and superoxide dismutase 1 G93A(SOD1-G93A),which fo rm abnormal aggregates in ALS.We found that the expression of these proteins increased following the administration of all three 5-HT receptor antagonists.In addition,the disease-related mislocalization of TD P-43 to the cytoplasm increased markedly for all three drugs.In ce rtain anatomical regions,the 5-HT receptor antagonists also led to a marked increase in the number of astrocytes and microglia and a decrease in the number of neurons.These results indicate that 5-HT deficiency may play a role in the pathogenesis of amyotrophic lateral sclerosis by inducing the abnormal expression and/or distribution of TDP-43 and SOD1-G93A and by activating glial cells.5-HT co uld therefore be a potential therapeutic target for amyotrophic lateral sclerosis.

Orbitofrontal cortex action of 5-hydroxytryptamine and its receptor in an acute forced swimming stress-induced depression model

BACKGROUND： The orbitofrontal cortex （OFC） is a brain region closely associated with emotion. 5-hydroxytryptamine （5-HT） has been shown to be involved in human depression. OBJECTIVE： To investigate OFC actions and mechanisms of 5-HT and 5-HT1A receptor （5-HT1AR） in stress-induced depression.DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at Laboratory of Neurobiology, College of Life Science, Shaanxi Normal University between May 2006 and March 2008. MATERIALS： 5-HT, p-chlorophenylalanine （PCPA, an inhibitor to tryptophan hydroxylase） and spiperone （5-HT1AR antagonist） were provided by Sigma, USA; rabbit anti-rat 5-HT1AR antibody was provided by Tianjin Haoyang Biological Manufacture. METHODS： A total of 40 male Sprague Dawley rats, aged 3 months, were randomly divided into five groups： control, model, 5-HT, spiperone ＋ 5-HT, and PCPA, with 8 rats in each group. Except for control group, rats in the other four groups were used to establish depression models by forced swimming for 15 minutes. At 30 minutes before forced swimming test, 0.5 pL of 5-HT （12.5 pg/pL）, PCPA （20 pg/pL）, spiperone （1.3 pg/pL） ＋ 5-HT （12.5 pg/pL, 10 minutes later）, and saline were respectively injected into the OFC of 5-HT, PCPA, spiperone ＋ 5-HT, and model groups, respectively. The control group received a saline microinjection into the OFC.MAIN OUTCOME MEASURES： Forced swimming and open field tests were employed to measure animal behaviors, and immunohistochemistry was used to analyze 5-HT1AR expression in the OFC, cingulate cortex, and piriform cortex. RESULTS： （1） Compared with the model group, 5-HT microinjection into the OFC prominently reduced immobility time in the forced swimming test and rearing in open field test （P 〈 0.05）; locomotion and grooming in open field test were increased, although there was no significance （P 〉 0.05）. Furthermore, following PCPA microinjection into the OFC （PCPA ＋ forced swimming stress）, immobility time in forced swimming test increased dramatically （P〈 0.01）, locomotion and rearing in open field test declined （P〈 0.05 and P 〈 0.01, respectively）. Compared with the 5-HT group, 5-HT1AR antagonist （spiperone ＋ 5-HT ＋ forced swimming stress） increased immobility time in forced swimming test （P 〈 0.01）, but decreased locomotion, rearing, and grooming in open field test. （2） Forced swimming stress markedly elevated 5-HT1AR expression in OFC, cingulate cortex, and piriform cortex （P〈 0.05 or P〈 0.01）. CONCLUSION： 5-HT improved stress-induced depression, and 5-HT anti-depression effects are primarily achieved via 5-HT1AR. Stress-induced up regulation of 5-HT1AR expression might be a compensatory mechanism for decreased 5-HT expression.

Role of 5-hydroxytryptamine expression in cerebellar Purkinje cells in obstructive sleep apnea syndrome

In the present study, electrical stimulation to the rat insular cortex induced apnea or respiratory disturbance, reduced amplitude of genioglossal electromyogram, and decreased electromyogram integrals. In addition, arterial blood gas analysis showed arterial blood acidosis, reduced pH values, increased alkali reserve negative values, decreased peripheral blood 5-hydroxytryptamine content, and increased 5-hydroxytryptamine expression in cerebellar Purkinje cells. Following lidocaine injection to block the habenular nucleus, abnormalities in breath, genioglossal electromyogram, and blood gas values disappeared, and peripheral blood 5-hydroxytryptamine content returned to levels prior to electric stimulation. However, 5-hydroxytryptamine expression in cerebellar Purkinje cells remained high. The results suggested that 5-hydroxytryptamine expression in Purkinje cells did not correlate with ventilation function involving insular cortex and habenular nucleus.

5-hydroxytryptamine-mediated apnea caused by the habenular nucleus

5-hydroxytryptamine contributes to the control of activities of the dilator muscle in the upper respiratory tract, and is derived from the raphe nuclei, in which the habenular nucleus exerts a sustaJned inhibitory effect. In the present study, respiratory motion curve of the genJoglossus muscle and peripheral 5-hydroxytryptamine changes were observed following L-glutamate stimulation of the habenular nucleus of adult Wistar rats. Results showed that the rats had apnea and decreased plasma 5-hydroxytryptamine content after the neurons in habenular nucleus were excited. Genioglossus muscle electromyogram amplitude and integral were significantly reduced. The genioglossus myoelectric activity and respiratory motion curve were similar to obstructive sleep apnea syndrome, thus confirming that the habenular nucleus is the key nucleus involved in the pathogenesis of obstructive sleep apnea syndrome, and is the primary regulated center in the raphe nuclei. Stimulation of the habenular nucleus may suppress 5-hydroxytryptamine release and result in apnea, which is similar to obstructive sleep apnea syndrome.

Inhibiting 5-hydroxytryptamine receptor 3 alleviates pathological changes of a mouse model of Alzheimer's disease

Extracellular amyloid beta(Aβ) plaques are main pathological feature of Alzheimer’s disease.However,the specific type of neuro ns that produce Aβ peptides in the initial stage of Alzheimer’s disease are unknown.In this study,we found that 5-hydroxytryptamin receptor 3A subunit(HTR3A) was highly expressed in the brain tissue of transgenic amyloid precursor protein and presenilin-1 mice(an Alzheimer’s disease model) and patients with Alzheimer’s disease.To investigate whether HTR3A-positive interneurons are associated with the production of Aβ plaques,we performed double immunostaining and found that HTR3A-positive interneurons were clustered around Aβ plaques in the mouse model.Some amyloid precursor protein-positive or β-site amyloid precursor protein cleaving enzyme-1-positive neurites near Aβ plaques were co-localized with HTR3A interneurons.These results suggest that HTR3A-positive interneurons may partially contribute to the generation of Aβ peptides.We treated 5.0-5.5-month-old model mice with tro pisetron,a HTR3 antagonist,for 8 consecutive weeks.We found that the cognitive deficit of mice was partially reversed,Aβ plaques and neuroinflammation we re remarkably reduced,the expression of HTR3 was remarkably decreased and the calcineurin/nuclear factor of activated T-cell 4 signaling pathway was inhibited in treated model mice.These findings suggest that HTR3A interneurons partly contribute to generation of Aβ peptide at the initial stage of Alzheimer’s disease and inhibiting HTR3 partly reve rses the pathological changes of Alzheimer’s disease.

白藜芦醇激活细胞外信号调节激酶5信号蛋白促进小鼠MC3T3-E1细胞增殖

背景:细胞外信号调节激酶5信号蛋白对生物体的存活不可或缺,白藜芦醇能通过多种途径促进成骨细胞增殖,但其是否能通过细胞外信号调节激酶5信号蛋白调控成骨细胞功能还需进一步验证。目的:探究细胞外信号调节激酶5对MC3T3-E1细胞增殖以及相关分泌蛋白的调控作用,进一步验证白藜芦醇通过激活细胞外信号调节激酶5完成上述过程。方法:小鼠MC3T3-E1前成骨细胞分别用完全培养基、XMD8-92(细胞外信号调节激酶5抑制剂)、表皮生长因子(细胞外信号调节激酶5激活剂)和白藜芦醇单独干预及XMD8-92+表皮生长因子、白藜芦醇+XMD8-92干预后,通过Western blot检测各组细胞内细胞外信号调节激酶5、磷酸化细胞外信号调节激酶5蛋白,增殖相关蛋白Cyclin D1、CDK4、PCNA,以及成骨细胞分泌蛋白骨保护素、核因子κB受体活化因子配体的表达情况,使用细胞免疫荧光染色检测各组细胞外信号调节激酶5、骨保护素和核因子κB受体活化因子配体荧光强度,使用EdU染色检测各组细胞增殖情况。白藜芦醇干预MC3T3-E1细胞的适宜浓度及时间由细胞形态学观察和CCK-8实验确定。结果与结论:①细胞外信号调节激酶5信号蛋白的激活能有效促进MC3T3-E1细胞增殖、上调骨保护素/核因子κB受体活化因子配体比值;②白藜芦醇干预MC3T3-E1细胞的适宜浓度及时间为5μmol/L,24 h;③白藜芦醇可以激活细胞外信号调节激酶5信号蛋白,进而促进成骨细胞增殖,并上调骨保护素/核因子κB受体活化因子配体比值;④研究结果表明,白藜芦醇可以通过激活细胞外信号调节激酶5信号蛋白促进MC3T3-E1细胞增殖,并通过激活细胞外信号调节激酶5信号蛋白上调骨保护素/核因子κB受体活化因子配体比值。

Synthesis and Metabolism of 5-hydroxytryptamine in Peripheral Circulatory System of Suckling Piglets

[ Objective] To find the rule of 5-hydroxytryptamine （5-HT） synthesis and metabolism in peripheral blood of suckling piglets with aging. [ Method ] All of 15 British Large White pigs at 0, 5 and 15 d of age were selected as experimental animals, and their serumal concentrations of L- tryptophan （L-Trp）, 5-HT and 5-hydroxyindoleacetic acid （5-HIAA） in peripheral circulatory system were determined synchronously by reversed- phase high-performance liquid chromatography method with UV. [ Result] The concentration of L-Trp was （16. 92 ± 1.74） iJmol/L at birth, （59.94 ±10.88） iJmol/L at 5 d of age and （70.42± 6.48） μmol/L at 15 d of age. The concentration of 5-HT was （ 12.85 ± 1.79） iJmol/L at birth, （4.81 ±2.05） ijmol/L at 5 d of age and （5.49±1.09） μmol/L at 15 d of age. While 5-HIAA, the metabolic product of 5-HT, was not detected at all the samples selected. [ Conclusion] The L-Trp concentration of suckling piglets increase significantly with aging （ P 〈0. 05）. While the concentra- tion of 5-HT and the conversion ratio of L-Trp to 5-HT are both highest at birth （P〈0.05） and keep stable at 5 and 15 d of age.

Changes of 5-hydroxytryptamine and dopamine contents in brain microvessels and the influence on cerebral edema at early stage of brain injury

The contents of 5-hydroxytryptamine(5-HT)and dopamine(DA) in the brain microvessels(BMVs) at the early stage of rat brain injury were measured by using high performance liquid chromatography with electrochemical detector(HPLC-ECD) and the influence of the

基于改进YOLOv5的红花目标检测算法研究

为实现农业非结构环境下采摘机器人对红花的准确识别,提出了一种基于改进YOLOv5的红花目标检测算法。将CBAM注意力机制嵌入到YOLOv5网络,提高了小尺寸目标物在高层次特征中的表现力;建立一种Alpha-IoU目标位置损失函数对原损失函数GIOU存在的梯度消失问题进行改进,提高了被遮挡红花的预测率,并通过在目标检测网络中增加分割检测模块,提高宽和高小于最低像素的小目标物检测精度,利用图像扩增数据集对改进后的YOLOv5算法进行训练,再分别与改进前后YOLOv5网络和Faster R-CNN网络在不同红花品种、不同自然光照情况、不同天气条件和不同遮挡情况下进行对比。试验结果表明:改进后的YOLOv5算法P值、R值分别为90.45%和0.90,对非结构环境下盛开期的未采摘红花mAP值达到94.48%,在不同影响因素下都可以准确识别出红花且置信度较高,可为红花采摘机器人自动化作业中的红花识别提供技术支持。

Effects of 5-hydroxytryptamine Receptor on the Reaction of the Lower Esophageal Sphincter Under the Electrical Field Stimulation

In the first and second parts of this study,5-hydroxytryptamine(5HT)receptors,including 5-HT3 and 5-HT4 with the highest expression level,were found in clasp and sling fibres of the lower esophageal sphincter(LES).Specific 5-HT3 and 5-HT4 receptor agonists can induce the contraction effect of clasp and sling fibres of the LES while specific 5-HT7 receptor agonists showed no effects.In the study of this part,the in-vitro muscle tension measurement technology and EFS methods were used to detect the effect of the selective 5-HT receptor antagonist on the clasp and sling fibres of the in-vitro LES under the electrical field stimulation(EFS),and further to ensure the effect of 5-HT receptor in the LES neuroregulatory pathway,and deeply explore the effect of 5-HT receptor in the systolic and diastolic function regulation of the LES.

过表达溶质载体家族1成员5和敲低慢病毒载体构建及稳定转染RAW264.7细胞株

背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供有力的实验工具。目的:构建小鼠SLC1A5过表达和敲低的慢病毒载体,以建立稳定转染的RAW264.7细胞株,为深入探讨SLC1A5在炎症中的作用提供实验基础。方法:根据SLC1A5基因序列设计合成引物并使用聚合酶链反应扩增该基因片段。将目的基因定向接入经Age I/Nhe I酶切的载体质粒GV492中构建重组慢病毒质粒,对阳性克隆进一步筛选后测序比对结果;pHelper1.0质粒载体、pHelper2.0质粒载体、目的质粒载体与293T细胞共同培养并转染,获得慢病毒原液进行包装和滴度测定;在此基础上,通过体外培养RAW264.7细胞,确定嘌呤霉素工作质量浓度;不同滴度的慢病毒分别与RAW264.7细胞共同培养,根据荧光强度确定转染效率;用嘌呤霉素挑选出稳定转染细胞,实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测稳定转染细胞株的SLC1A5基因和蛋白表达水平。结果与结论:(1)测序序列与目的序列一致提示重组慢病毒载体构建成功;(2)过表达SLC1A5慢病毒的滴度为1×10~9 TU/mL,敲低SLC1A5慢病毒的滴度为3×10~9 TU/mL;(3)确定RAW264.7细胞嘌呤霉素工作质量浓度为3μg/mL;(4)过表达/敲低SLC1A5慢病毒转染RAW264.7细胞的最佳条件皆为HiTransG P转染增强液且感染复数值等于50;(5)过表达SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量明显上调,而敲低SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量显著下调。结果表明,成功构建了小鼠SLC1A5过表达和敲低的慢病毒载体并获得稳定转染的RAW264.7细胞株。

5-羟甲基糠醛在CuPd(111)上的加氢反应机理研究

采用密度泛函理论,利用包含零点能校正的PBE-D3方法研究了5-羟甲基糠醛在双金属催化剂CuPd(111)上表面的加氢还原反应,考虑了两种反应路径,第一种反应路径是侧链上的醛基氧首先发生加氢反应生成F-CHOH中间体,然后F-CHOH中间体继续加氢生成2,5-二羟甲基呋喃;第二种反应路径是侧链上的醛基碳先加氢原还原生成F-CH_(2)O中间体,生成的F-CH_(2)O进一步加氢还原生成2,5-二羟甲基呋喃.计算了两种竞争反应路径的活化能垒和反应热,结果表明,5-羟甲基糠醛在CuPd(111)面加氢还原生成2,5-二羟甲基呋喃的最佳反应路径为:F-CHO→F-CHOH→F-CH_(2)OH.

miR-192-5p在增生性瘢痕组织及成纤维细胞中的表达及调控

背景:研究表明,miRNAs的表达与肝纤维化、肾纤维化及皮肤纤维化发生有关;并证实在痛风性关节炎中miR-192-5p与表皮调节素存在靶向调控关系。目的:探讨miR-192-5p在增生性瘢痕中的表达及调控作用,并验证miR-192-5p与表皮调节素之间存在靶向调控关系。方法:①在新疆医科大学第一附属医院收集6例增生性瘢痕组织及6例正常皮肤组织,采用qRT-PCR法检测miR-192-5p与表皮调节素mRNA表达。②组织块法获取原代增生性瘢痕成纤维细胞,传代培养至3-6代用于后续实验,实验分为3个组:阴性对照组、miR-192-5p模拟物组和miR-192-5p抑制物组,分别转染对应的序列。采用CCK-8法和EdU试剂盒检测细胞增殖活力,划痕实验检测细胞迁移能力,流式细胞术检测细胞凋亡,qRT-PCR和Western blot法检测表皮调节素、Ⅰ型胶原、Ⅲ型胶原和α-SMA的基因和蛋白表达,生物信息学网站预测miR-192-5p靶点,双荧光素酶实验验证靶向结合。结果与结论:①与正常皮肤组织及其成纤维细胞相比,miR-192-5p和表皮调节素在增生性瘢痕和增生性瘢痕成纤维细胞中均呈高表达(P<0.05或P<0.01);②过表达miR-192-5p后,与阴性对照组比较,细胞增殖能力增强(P<0.05),EdU阳性细胞率增加(P<0.01);抑制miR-192-5p后细胞活力降低(P<0.05),EdU阳性细胞率减少(P<0.05);③过表达miR-192-5p 24 h后,与阴性对照组比较,模拟物组细胞划痕间面积、细胞凋亡率减小(P<0.05),miR-192-5p抑制物组细胞划痕间面积、细胞凋亡率增加(P<0.01);④转染48 h后,miR-192-5p模拟物组表皮调节素mRNA及蛋白水平显著降低、Ⅰ型胶原、Ⅲ型胶原和α-平滑肌肌动蛋白mRNA及蛋白水平显著增高(P<0.05或P<0.01);miR-192-5p抑制物组上述4项指标呈现相反的变化(P<0.05或P<0.01);⑤Targetscan网站预测表皮调节素与miR-192-5p有潜在结合位点;⑥双荧光素酶实验显示,miR-192-5p能够与表皮调节素靶向结合。结果说明:miR-192-5p过表达可降低表皮调节素的表达,二者可能存在负向调控;提示通过调控表皮调节素可能起到抑制增生性瘢痕成纤维细胞增殖的作用。

经“O”点穿刺与传统单侧入路治疗L_(5)椎体骨质疏松性压缩性骨折

背景:单侧穿刺入路经皮穿刺球囊扩张椎体后凸成形治疗骨质疏松性椎体压缩骨折优于双侧穿刺入路,由于L_(5)特殊解剖形态和位置特殊,传统单侧穿刺入路治疗L_(5)椎体骨质疏松性椎体压缩骨折适用性差,因此提出“O”点穿刺入路来解决该问题。目的:比较经横突基底部-椎弓根后上部-关节突外侧交点(“O”点)入路与传统单侧入路经皮穿刺球囊扩张椎体后凸成形治疗L_(5)椎体骨质疏松性压缩骨折的疗效差异。方法:回顾性分析西南医科大学附属医院2020年1月至2022年12月收治的54例L_(5)椎体骨质疏松性压缩骨折行经“O”点入路或传统经皮穿刺球囊扩张椎体后凸成形治疗的患者,根据手术方式分为2组,“O”点入路组(A组)29例,传统入路组(B组)25例。术前在X射线正位片测量A组“O”点与髂棘位置及穿刺角度;比较两组术前与术后第2天、末次随访的局部Cobb角、椎体前缘、中部高度及骨水泥分布情况;术前、术后第2天及末次随访采用疼痛目测类比评分、Oswestry功能障碍指数评估患者疼痛缓解情况与日常生活能力,记录手术相关并发症。结果与结论:①A组手术时间、术中透视次数均少于B组,差异有显著性意义(P<0.05);两组术中出血量及骨水泥注射量差异无显著性意义;②两组患者术后第2天及末次随访疼痛目测类比评分、Oswestry功能障碍指数较术前明显降低,且A组降低比B组更明显(P<0.05);③“O”点位于双侧髂棘最高点连线之下平均距离1.23 cm;“O”点到髂棘平均横向距离2.89 cm;④两组患者末次随访Cobb角较术前明显改善(P<0.05),但两组间比较差异无显著性意义(P>0.05);⑤术后骨水泥分布,A组分布良好占97%(28/29),B组分布良好占88%(22/25),A组明显优于B组(P<0.05);⑥提示经“O”点穿刺入路与传统入路经皮穿刺球囊扩张椎体后凸成形治疗L_(5)椎体骨质疏松性压缩骨折疗效满意,但“O”点穿刺入路经皮穿刺球囊扩张椎体后凸成形治疗骨水泥分布更满意,还可避免高髂棘对L_(5)椎体穿刺入路的影响。

Ovarian hormone modulates 5-hydroxytryptamine 3 receptors mRNA expression in rat colon with restraint stress-induced bowel dysfunction

AIM: To examine the effects of ovarian hormone on the expression of 5-hydroxytryptamine 3 receptors (5-HT3R) in rat colon of restraint stress-induced bowel dysfunction.METHODS: Twenty-four female Sprague-Dawley rats were randomly divided into three groups of 8 each: sham operation,ovariectomy (OVX) and ovariectomy with estrogen (E2) and progesterone (P) replacement therapy (OVX+E2+P). The rats were subjected to 1-h restraint stress 4 wk after operation.The changes of defecation were monitored by collection of fecal pellets. The gonadal steroids were measured in duplicate by radioimmunoassay (RIA). The expression of 5-HT3R mRNA in the colon was studied by RT-PCR.RESULTS: Compared with sham group and OVX+E2+P group, OVX group showed increase in fecal pellets and decrease in the time of vitreous pellets excretion (P<0.01).Serum levels of E2 and P were suppressed in OVX group and restored following treatment with ovarian steroids (P<0.01),and the levels of 5-HT3R mRNA in the colon of ovariectomized rats were significantly increased, the expression of 5-HT3R mRNA was significantly decreased in hormone replacement therapy group (P<0.01).CONCLUSION: Ovarian hormone plays a role in the regulation of 5-HT3R expressions in restraint stress-induced bowel dysfunction of rats. The interactions between ovarian steroids and gastrointestinal tract may have major pathophysiological implications in 5-HT-related disorders, such as irritable bowel syndrome (IBS).

Mechanism by which Rab5 promotes regeneration and functional recovery of zebrafish Mauthner axons

Rab5 is a GTPase protein that is involved in intracellular membrane trafficking. It functions by binding to various effector proteins and regulating cellular responses, including the formation of transport vesicles and their fusion with the cellular membrane. Rab5 has been reported to play an important role in the development of the zebrafish embryo;however, its role in axonal regeneration in the central nervous system remains unclear. In this study, we established a zebrafish Mauthner cell model of axonal injury using single-cell electroporation and two-photon axotomy techniques. We found that overexpression of Rab5 in single Mauthner cells promoted marked axonal regeneration and increased the number of intra-axonal transport vesicles. In contrast, treatment of zebrafish larvae with the Rab kinase inhibitor CID-1067700markedly inhibited axonal regeneration in Mauthner cells. We also found that Rab5 activated phosphatidylinositol 3-kinase(PI3K) during axonal repair of Mauthner cells and promoted the recovery of zebrafish locomotor function. Additionally, rapamycin, an inhibitor of the mechanistic target of rapamycin downstream of PI3K, markedly hindered axonal regeneration. These findings suggest that Rab5 promotes the axonal regeneration of injured zebrafish Mauthner cells by activating the PI3K signaling pathway.

酸枣仁提取物调控miR-7b-3p/5-羟色胺1A受体表达促进骨生长

背景:前期研究发现,酸枣仁提取物通过提高脑组织5-羟色胺1A受体(serotonin 1A receptor,5-HT1AR)表达,使之与5-羟色胺结合,延长小鼠慢波睡眠,促进生长激素的分泌,从而使骨生长。5-HT1AR作为一种蛋白质,其表达丰度受到miRNA的调控。作者推测酸枣仁提取物可能通过miRNA调控5-HT1AR的表达从而发挥药物作用。目的:观察酸枣仁提取物通过干预小鼠脑组织中miR-7b-3p/5-HT1AR通路对骨骼生长的影响。方法:①将昆明种小鼠分为正常对照组、用药组(灌胃酸枣仁提取物0.320 mg/g),阳性对照组(灌胃酸枣仁皂甙0.013 mg/g)和酸枣仁提取物+5-HT1AR抑制剂组(灌胃酸枣仁提取物最后3 d同时每天侧脑室注射5-HT1AR选择性抑制剂P-MPPF 8μg),25 d后观察酸枣仁提取物对小鼠骨生长、血清生长激素水平及脑组织5-HT1AR表达的影响;②基因芯片法筛选由酸枣仁提取物引起的骨生长小鼠与普通小鼠脑组织差异表达的miRNAs,并通过PCR验证和双荧光素酶报告基因实验证实筛选的miR-7b-3p与5-HT1AR的调控关系;③体外培养小鼠脑皮质细胞并鉴定,利用Western blot法观察酸枣仁提取物对脑皮质细胞中5-HT1AR表达的影响;④将昆明种小鼠分为正常对照组、用药组、miR-7b-3p inhibitor组、用药+miR-7b-3p mimics组、阳性对照组,观察各组小鼠脑组织5-HT1AR表达及5-羟色胺与5-HT1AR结合活性;⑤将SD大鼠分为正常对照组、用药组、miR-7b-3p inhibitor组、用药+miR-7b-3p mimics组、阳性对照组,观察各组大鼠慢波睡眠的变化。结果与结论:①酸枣仁提取物可以促进小鼠体长、胫骨增长,促进生长激素分泌,提高脑组织5-HT1AR含量;②基因芯片筛选出差异表达的miRNAs个数为16个,其中上调有13个,下调有3个;生物信息学预测下调miR-7b-3p可以调控5-HT1AR表达,且双荧光素酶报告基因实验证实二者有直接调控关系;③酸枣仁提取物和沉默脑皮质细胞中miR-7b-3p表达可以引起5-HT1AR高表达;沉默miR-7b-3p后小鼠脑组织5-HT1AR表达、5-羟色胺与5-HT1AR结合活性及生长激素分泌均升高;过表达miR-7b-3p后小鼠脑组织5-HT1AR表达、5-羟色胺与5-HT1AR结合活性及生长激素分泌均降低;大鼠慢波睡眠期也相应延长或缩短。结果表明,酸枣仁提取物能够降低脑组织的miR-7b-3p水平,同时增加5-HT1AR的表达量。这种机制有助于延长慢波睡眠周期,并且促进生长激素的生成,对骨骼生长有积极影响,为使用酸枣仁提取物作为促进骨骼生长的潜在手段提供了科学依据。