Enriched environment elevates expression of growth associated protein-43 in the substantia nigra of SAMP8 mice

An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43（GAP-43） is closely associated with neurite outgrowth and axon regeneration during neural development. We speculate that an enriched environment can reduce damage to dopaminergic neurons by affecting the expression of GAP-43. This study is designed to test this hypothesis. Three-month-old female senescence-accelerated mouse prone 8（SAMP8） mice were housed for 3 months in an enriched environment or a standard environment. These mice were then subcutaneously injected in the abdomen with 14 mg/kg MPTP four times at 2-hour intervals. Morris water maze testing demonstrated that learning and memory abilities were better in the enriched environment group than in the standard environment group. Reverse-transcription polymerase chain reaction, immunohistochemistry and western blot assays showed that m RNA and protein levels of GAP-43 in the substantia nigra were higher after MPTP application in the enriched environment group compared with the standard environment group. These findings indicate that an enriched environment can increase GAP-43 expression in SAMP8 mice. The upregulation of GAP-43 may be a mechanism by which an enriched environment protects against MPTP-induced neuronal damage.

Pathological changes in the retina and growth associated protein-43 expression following treatment of intracanalicular optic nerve injury via optic canal decompression,dexamethasone or their combination

BACKGROUND： The main clinical treatments for optic nerve injury are optic canal decompression and systemic administration of hormones, but both treatments have disadvantages. OBJECTIVE： To observe the pathological changes in the retina and growth associated protein-43 （GAP-43） expression, to compare the treatment of optic canal decompression, hormones, and their combination with the intracanalicular optic nerve injury.DESIGN, TIME AND SETTING： A randomized, controlled animal study was performed at the Department of Anatomy, Weifang Medical University, China, from September 2007 to November 2008.MATERIALS： Dexamethasone （Shandong Huaxin Pharmaceutical, China） and rabbit anti-GAP-43 polyclonal antibody （Boster, China） were used.METHODS： All 36 healthy adult rabbits were randomly assigned to control group （n = 4）, simple injury group （n = 20）, and treatment group （n = 12）. Intracanalicular optic nerve injury models were established using the metal cylinder free-fall impact method. The control group was left intact. The treatment group （four rabbits in each subgroup） was treated by optic nerve decompression, dexamethasone treatment （1 mg/kg daily via two intravenous infusions, 1/5 total dose reduction every 3 days, for 14 days）, and simultaneously giving surgery and hormone treatment.MAIN OUTCOME MEASURES： Pathological changes in the retina were determined using hematoxylin-eosin staining. GAP-43 expression was detected using immunohistochemistry in the retina.RESULTS： Retina injury induced obvious pathological changes in the retina. With prolonged time after optic nerve injury, the number of retinal ganglion cells was gradually decreased, and reached the minimum on day 14 （P〈0.01）. All three treatments increased the number of retinal ganglion cells （P〈0.01）, but surgery ＋ hormone treatment was most effective. No GAP-43 cells were present in the normal retinal, but they appeared 3 days after injury, peaked 7 days after injury, and then began to decline.CONCLUSION： Intracanalicular optic nerve injury induced obvious pathological changes in the retina, including increased GAP-43 expression. Optic canal decompression and hormones improved nerve repair after injury, and their combination produced better outcomes.

Effects of continuous peripheral nerve block by tetrodotoxin on growth associated protein-43 expression during neuropathic pain development

BACKGROUND： Peripheral nerve injury may lead to neuropathic pain and cause a markedly increase expression of growth associated protein-43 （GAP-43） in the spinal cord and dorsal root ganglion, local anesthetics blocking electrical impulse propagation of nerve fibers may also affect the expression of GAP-43 in the spinal cord and dorsal root ganglion. OBJECTIVE： To determine the effects of continuous peripheral nerve block by tetrodotoxin before and after nerve injury on GAP-43 expression in the dorsal root ganglion during the development of neuropathic pain. DESIGN： A randomized controlled animal experiment. SETTINGS： Department of Anesthesiology, the Second Hospital of Xiamen City; Department of Anesthesiology, the Second Affiliated Hospital of Shantou University Medical College. MATERIALS： Thirty-five Spragne Dawley （SD） rats, weighing 200 - 250 g, were randomly divided into four groups： control group （n =5）, simple sciatic nerve transection group （n =10）, peripheral nerve block before and after sciatic nerve transection groups （n =10）. All the sciatic nerve transection groups were divided into two subgroups according to the different postoperative survival periods： 3 and 7 days （n =5） respectively. Mouse anti-GAP-43 monoclonal antibody （Sigma Co., Ltd.）, supervision TM anti-mouse reagent （HRP, Changdao antibody diagnosis reagent Co., Ltd., Shanghai）, and HMIAS-100 image analysis system （Qianping Image Engineering Company, Tongji Medical University） were employed in this study. METHODS： This experiment was carried out in the Department of Surgery and Pathological Laboratory, the Second Affiliated Hospital of Shantou University Medical College from April 2005 to April 2006. ①The animals were anesthetized and the right sciatic nerve was exposed and transected at 1 cm distal to sciatic notch. ② Tetrodotoxin 10 μg/kg was injected percutaneously between the greater trochanter and the posterior superior iliac spine of fight hind limb to block the sciatic nerve proximally at 1 hour before or 4 hours after nerve injury respectively, the injection was repeated in all the rats every 12 hours.③ At 3 or 7 days after nerve injury, immunohistochemistry and image analysis were used to evaluate the expression of GAP-43 in the dorsal root ganglions of L5 to the transected sciatic nerve, and quantitative analysis was also performed. ④ Statistical analysis was performed using one way analysis of variance followed by t test. MAIN OUTCOME MEASURE： Expression of GAP-43 in the fight dorsal root ganglions of L5. RESULTS： All the 35 SD rats were involved in the final analysis of results. In normal rats, there were very low expressions of GAP-43 in the dorsal root ganglions. In simple sciatic nerve transection rats 3 and 7 days after sciatic nerve transection, the average absorbance value of GAP-43 immunopositive neurons were significantly different from that in normal rats （t =8.806, 6.771, P 〈 0.01）. Whereas 3 and 7 days after sciatic nerve transection in rats with peripheral nerve block before and after nerve injury, the average absorbance value of GAP-43 immunopositive neurons were not significantly different from that in normal rats （P 〉 0.05）. CONCLUSION： Local anesthetic continuous peripheral nerve block before or after nerve injury can suppress nerve injury induced high expression of GAP-43 during the development of neuropathic pain.

Neurofilament 200 expression in a rat model of complete spinal cord injury following growth-associated protein-43 treatment

BACKGROUND： Growth-associated protein-43 （GAP-43） expression in the nervous system has been demonstrated to promote neural regeneration, neuronal growth and development, as well as synaptic reconstruction. Neurofilament 200 （NF200） expression could reflect degree of injury and repair in injured spinal axons. OBJECTIVE： To observe NF200 expression changes in a rat model of complete spinal cord injury following GAP-43 treatment and to explore the effects of GAP-43 following spinal cord injury. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Laboratory of Histology and Embryology of Kunming Medical University between March 2007 and October 2008. MATERIALS： GAP-43 and GAP-43 antibody were provided by Beijing Boao Biology, China; mouse anti-rat NF200 antibody was purchased from Chemicon, USA. METHODS： Female, 8-week-old, Sprague Dawley rats were randomly assigned into three groups following complete spinal cord injury, with 20 animals in each group： GAP-43 antibody, GAP-43, and model groups. In addition, each group was subdivided into four subgroups according to sampling time after modeling, Le., 3-, 5-, 9-, and 15-day groups, with 5 rats in each group. GAP-43 antibody or GAP-43 was injected into injury sites of the spinal cord, 5 μg/0.2 mL, respectively, twice daily for three consecutive days, followed by three additional days of injection, once daily. The model group did not receive any treatment following injury. MAIN OUTCOME MEASURES： NF200 expression in the damaged spinal area at different stages was detected by immunohistochemistry; lower limb motion function following injury was evaluated using the Basso, Beattie and Bresnahan （BBB） locomotor rating scale. RESULTS： NF200 expression was significantly reduced in the GAP-43 antibody group, compared with GAP-43 and model groups, at 3 and 5 days after spinal cord injury （P 〈 0.05）. In addition, the model group expressed significantly less NF200 than the GAP-43 group （P 〈 0.05）. BBB scores from the GAP-43 antibody and model groups were remarkably less than the GAP-43 group （P 〈 0.05）. At 9 and 15 days of injury after drug withdrawal, NF200 expression was increased in the GAP-43 antibody group, and NF200 expression and BBB scores in the GAP-43 antibody and GAP-43 groups were significantly greater than in the model group （P 〈 0.05）. In particular, the GAP-43 group exhibited greater BBB scores than the GAP-43 antibody group at day 9 （P 〈 0.05）. CONCLUSION： GAP-43 promoted NF200 expression and recovery of lower limb function. Early administration of GAP-43 antibody produced reversible nerve inhibition, which was rapidly restored following withdrawal.

Peripheral nerve regeneration through nerve conduits evokes differential expression of growth-associated protein-43 in the spinal cord

Growth-associated protein 43 plays a key role in neurite outgrowth through cytoskeleton remodeling.We have previously demonstrated that structural damage of peripheral nerves induces growth-associated protein 43 upregulation to promote growth cone formation.Conversely,the limited regenerative capacity of the central nervous system due to an inhibitory environment prevents major changes in neurite outgrowth and should be presumably associated with low levels of growth-associated protein 43 expression.However,central alterations due to peripheral nerve damage have never been assessed using the growthassociated protein 43 marker.In this study,we used the tubulization technique to repair 1 cm-long nerve gaps in the rat nerve injury/repair model and detected growth-associated protein 43 expression in the peripheral and central nervous systems.First,histological analysis of the regeneration process confirmed an active regeneration process of the nerve gaps through the conduit from 10 days onwards.The growth-associated protein 43 expression profile varied across regions and follow-up times,from a localized expression to an abundant and consistent expression throughout the regeneration tissue,confirming the presence of an active nerve regeneration process.Second,spinal cord changes were also histologically assessed,and no apparent changes in the structural and cellular organization were observed using routine staining methods.Surprisingly,remarkable differences and local changes appeared in growth-associated protein 43 expression at the spinal cord level,in particular at 20 days post-repair and beyond.Growth-associated protein 43 protein was first localized in the gracile fasciculus and was homogeneously distributed in the left posterior cord.These findings differed from the growth-associated protein 43 pattern observed in the healthy control,which did not express growth-associated protein 43 at these levels.Our results revealed a differential expression in growth-associated protein 43 protein not only in the regenerating nerve tissue but also in the spinal cord after peripheral nerve transection.These findings open the possibility of using this marker to monitor changes in the central nervous system after peripheral nerve injury.

Effects of Continuous Sciatic Nerve Block by Tetrodotoxin on Growth Associated Protein-43 Expression in Dorsal Root Ganglions of Normal and Sciatic Nerve Injury Rats

Growth associated protein-43 （GAP-43） is considered to be one of the most useful molecular markers for the neural development, nerve regeneration, and neuroplasticity. In most mature neurons, the expression of GAP-43 is at very low or negative level; its expression is triggered in response to the interruption of axonal transport. The purpose of this study was to examine whether continuous sciatic nerve block by tetrodotoxin （TTX） affects GAP-43 expression in the dorsal root ganglion （DRG） of normal and sciatic nerve injury rats.

Correlations of the expression of Cx43,SCFFBXW7,p-cyclin E1(Ser73),p-cyclin E1(Thr77)and p-cyclin E1(Thr395)in colon cancer tissues

BACKGROUND Previous cellular studies have demonstrated that elevated expression of Cx43 promotes the degradation of cyclin E1 and inhibits cell proliferation through ubiquitination.Conversely,reduced expression results in a loss of this capacity to facilitate cyclin E degradation.The ubiquitination and degradation of cyclin E1 may be associated with phosphorylation at specific sites on the protein,with Cx43 potentially enhancing this process by facilitating the phosphorylation of these critical residues.AIM To investigate the correlation between expression of Cx43,SKP1/Cullin1/F-box(SCF)FBXW7,p-cyclin E1(ser73,thr77,thr395)and clinicopathological indexes in colon cancer.METHODS Expression levels of Cx43,SCFFBXW7,p-cyclin E1(ser73,thr77,thr395)in 38 clinical colon cancer samples were detected by immunohistochemistry and were analyzed by statistical methods to discuss their correlations.RESULTS Positive rate of Cx43,SCFFBXW7,p-cyclin E1(Ser73),p-cyclin E1(Thr77)and p-cyclin E1(Thr395)in detected samples were 76.32%,76.32%,65.79%,5.26%and 55.26%respectively.Positive expressions of these proteins were not related to the tissue type,degree of tissue differentiation or lymph node metastasis.Cx43 and SCFFBXW7(r=0.749),p-cyclin E1(Ser73)(r=0.667)and p-cyclin E1(Thr395)(r=0.457),SCFFBXW7 and p-cyclin E1(Ser73)(r=0.703)and p-cyclin E1(Thr395)(0.415)were correlated in colon cancer(P<0.05),and expressions of the above proteins were positively correlated in colon cancer.CONCLUSION Cx43 may facilitate the phosphorylation of cyclin E1 at the Ser73 and Thr195 sites through its interaction with SCFFBXW7,thereby influencing the ubiquitination and degradation of cyclin E1.

不同施肥处理对黑河43农艺性状、产量及品质的影响

为促进黑龙江省北部大豆产业提质增效,本研究以目前国内种植面积最大的大豆品种黑河43为研究对象,设置8组不同施肥处理:常规施肥区,尿素2 kg·666.67 m^(-2)、磷酸二铵8 kg·666.67 m^(-2)、硫酸钾5 kg·666.67 m^(-2);磷处理1区,磷酸二铵8 kg·666.67 m^(-2)、硫酸钾5 kg·666.67 m^(-2);磷处理2区,磷酸二铵10 kg·666.67 m^(-2)、硫酸钾5 kg·666.67 m^(-2);磷处理3区,磷酸二铵12 kg·666.67 m^(-2)、硫酸钾5 kg·666.67 m^(-2);无钾区,尿素2 kg·666.67 m^(-2)、磷酸二铵8 kg·666.67 m^(-2);无磷区,尿素5.13 kg·666.67 m^(-2)、硫酸钾5 kg·666.67 m^(-2);无氮区,磷酸钙8 kg·666.67 m^(-2)、硫酸钾5 kg·666.67 m^(-2);空白区。研究轮作条件下氮、磷、钾对黑河43农艺性状、品质及产量的影响。结果表明:不同肥料处理对黑河43农艺性状影响存在一定差异,其中氮肥施用与株高、底荚高显著正相关,其他性状差异均不显著;肥料处理对黑河43籽粒大小及品质的影响差异显著,其中空白CK与缺氮PK处理区百粒重、蛋白质含量均显著低于其它处理,缺氮PK处理区脂肪含量均显著高于其他处理;在合理范围内增施氮肥、磷肥可显著提高黑河43的产量,补充钾肥可显著提高黑河43抗病性进而提高产量,其中N3P3K处理区2年产量均显著高于其他处理区。缺素处理区2年产量结果一致,效应依次:NK>NP>PK;通过相关性聚类热图分析,将黑河43综合农艺性状分成2大类,第Ⅰ类为蛋白与产量构成因子:包含产量、单株粒重、单株粒数、单株荚数、节数、感病率、蛋白、底荚高、百粒重、株高,相邻性状两两相关性最高,第Ⅱ类为脂肪与密度,脂肪含量与密度显著正相关。将不同肥料处理分成2大类,第I类是缺N区,包含空白CK处理区和PK缺氮区,第Ⅱ类是含氮肥区,进一步说明氮肥是黑河43生物量形成的基础,平衡施肥对黑河43产量及品质具有重要促进作用。

抗旱丰产春小麦新品种陇春43号选育报告

为甘肃省小麦生产提供抗旱、抗病、丰产、优质小麦新品种,以促进小麦更新换代,实现小麦稳产增产,以衡7728为母本、陇春27号为父本进行了有性杂交,通过异地生态选择、采用系谱法成功选育出了抗旱丰产春小麦新品种陇春43号。2018—2019年参加甘肃省旱地春小麦区域试验,平均折合产量3074.70 kg/hm^(2),较对照品种西旱2号增产10.02%。2020年参加甘肃省旱地春小麦生产试验,平均折合产量3582.61 kg/hm^(2),较对照品种西旱2号增产9.81%。该品种具有高产稳产,优质,株型紧凑,抗旱,抗倒伏,中抗条锈和白粉病等特点,适宜在甘肃省中部旱地春麦区以及类似生态地区种植。

基于GR/ERK/CX43研究母代肾精亏虚诱发子代原发性睾丸生精功能减弱的宫内编程机制

目的明确母代肾精亏虚导致子代原发性睾丸生精功能减弱的宫内编程机制。方法制备孕鼠24只,数字随机法均分为空白组,模型组,补肾组;除空白组以外,余下各组孕期采用慢性应激复制母鼠肾精亏虚模型。酶联免疫吸附法(ELISA)法检测母鼠血清甲状腺素(T4)、糖皮质激素(GC)、胎产数评估母鼠模型。从孕0天开始,补肾组给予左归丸灌胃填补肾精;孕期20天时,比较母鼠一般情况及体重变化,雄性胎鼠睾丸体质比,ELISA法检测其血清T4、GC、促卵泡生长激素(FSH)含量,并通过苏木素-伊红染色评估睾丸生殖细胞发育状况,免疫荧光双染检测胎鼠睾丸缝隙连接蛋白43(Cx43)、SYR-盒包含蛋白9(Sox9)表达,实时荧光定量聚合酶链式反应检测胎鼠睾丸细胞外调节蛋白激酶(ERK1/2)、Cx43基因表达,蛋白免疫印迹法检测胎盘组织GR、胎鼠睾丸ERK1/2、Cx43、促卵泡生长激素受体(FSHR)蛋白表达。结果与空白组母鼠比较,模型组母鼠,产后体重减轻、胎产数减少、血清T4含量降低、血清GC含量升高(P<0.01);与模型组母鼠比较,补肾组母鼠,产后体重增加、胎产数增加、血清T4含量升高、GC降低、(P<0.01)。与空白组雄性胎鼠比较,模型组雄性胎鼠血清T4含量降低、GC、FSH含量均升高、睾丸体质比降低、支持细胞数量、精原细胞数量及曲精小管个数均降低、睾丸Sox9、Cx43蛋白降低,ERK、Cx43 mRNA表达降低、FSHR表达升高(P<0.01或P<0.05),胎盘组织GR蛋白升高(P<0.05);与模型组比较,补肾组胎鼠血清T4含量升高、GC、FSH含量均降低、睾丸体质比升高、支持细胞数量、精原细胞数量及曲精小管个数均升高、睾丸ERK、Cx43 mRNA表达升高、Sox9、Cx43蛋白升高、FSHR表达降低(P<0.01或P<0.05),胎盘组织GR蛋白降低(P<0.05)。结论母鼠孕期肾精亏虚是原发性睾丸生精功能障碍的诱因之一,其机制可能与GR/ERK/CX43宫内编程调控支持细胞数量相关。

乌司他丁通过上调Cx43表达减轻缺氧/复氧诱导的心肌细胞铁死亡的作用机制

目的探讨乌司他丁(UTI)对缺氧/复氧(H/R)诱导的心肌细胞铁死亡的影响,并分析其保护机制。方法体外培养大鼠H9c2心肌细胞,分为对照组、H/R组、铁死亡诱导剂(Erastin)组、UTI组、UTI+Erastin组、UTI+小干扰RNA(siRNA)阴性对照(si-NC)组、UTI+连接蛋白43(Cx43)siRNA(si-Cx43)组,各组给予相应干预后,检测各组细胞活力、乳酸脱氢酶(LDH)活性、活性氧、Fe^(2+)、丙二醛、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及Cx43、谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链1(FTH1)、酰基辅酶A合成酶长链家族成员4(ACSL4)mRNA和蛋白水平。结果与对照组比较,H/R组细胞活力、SOD、CAT活性和Cx43、GPX4、FTH1mRNA及蛋白水平显著降低,LDH活性、活性氧、丙二醛、Fe^(2+)和ACSL4mRNA及蛋白水平显著升高(P<0.01)。与H/R组比较,UTI组细胞活力[(71.40±8.05)%vs(42.63±6.71)%]、SOD、CAT活性和Cx43、GPX4、FTH1mRNA及蛋白水平显著升高,LDH活性、活性氧、丙二醛、Fe^(2+)和ACSL4mRNA及蛋白水平显著降低(P<0.05)。与UTI组比较,UTI+Erastin组和UTI+si-Cx43组细胞活力、SOD、CAT活性和Cx43、GPX4、FTH1mRNA及蛋白水平显著降低,LDH活性、活性氧、丙二醛、Fe^(2+)和ACSL4mRNA及蛋白水平显著升高(P<0.05)。结论UTI可能通过上调Cx43抑制H/R诱导的心肌细胞铁死亡。

稳定转染TDP-43导致NSC34细胞自噬异常

目的 探讨稳定转染TDP-43对NSC34细胞自噬的影响。方法 体外培养稳定转染空质粒的NSC34细胞系(E细胞)和转染TDP-43的NSC34细胞系(TDP-43细胞)。应用CCK-8试剂盒测定两组细胞活力,应用EdU试剂盒测定两组细胞增值能力,应用透射显微镜观察两组细胞线粒体形态,应用免疫荧光检测两组细胞自噬相关蛋白表达水平。结果 与E组细胞相比,TDP-43组细胞活力明显下降,增殖能力下降,差异有统计学意义(P<0.05);线粒体形态异常,自噬相关蛋白表达水平升高,差异有统计学意义(P <0.05)。结论 稳定转染TDP-43可导致NSC34细胞自噬异常,线粒体形态改变,降低细胞活力和增殖能力。

血清Cx43、Gal-9水平对老年急性脑梗死患者超早期静脉溶栓治疗预后的评估价值

目的探讨血清间隙连接蛋白43(Cx43)、半乳糖凝集素-9(Gal-9)水平对老年急性脑梗死(ACI)患者超早期静脉溶栓治疗预后的评估价值。方法选取2020年9月至2022年9月在该院行超早期静脉溶栓治疗的106例老年ACI患者作为研究组,另外选取同期来该院体检的100例体检健康者作为健康组。采用酶联免疫吸附试验(ELISA)检测所有研究对象的血清Cx43、Gal-9水平。治疗2周后参考美国国立卫生院卒中量表(NIHSS)评分将106例老年ACI患者分为预后良好组(81例)与预后不良组(25例)。采用受试者工作特征(ROC)曲线分析血清Cx43、Gal-9水平对老年ACI患者超早期静脉溶栓治疗预后的评估价值,采用多因素Logistic回归分析探讨老年ACI患者超早期静脉溶栓治疗预后不良的影响因素。结果研究组血清Cx43、Gal-9水平高于健康组(P<0.05);预后不良组血清Cx43、Gal-9水平高于预后良好组(P<0.05);血清Cx43、Gal-9预测老年ACI患者预后不良的曲线下面积(AUC)分别为0.721(95%CI:0.673~0.758)、0.837(95%CI:0.787~0.886),二者联合预测的AUC为0.901(95%CI:0.857~0.946)。预后不良组高血压占比高于预后良好组(P<0.05);多因素Logistic回归分析显示,高血压(OR=3.487,95%CI:1.564~7.773),血清Cx43≥106.53 pg/mL(OR=4.586,95%CI:1.982~10.611),血清Gal-9≥11.84 ng/mL(OR=4.345,95%CI:1.957~9.648)是老年ACI患者超早期静脉溶栓治疗预后不良的危险因素(P<0.05)。结论血清Cx43、Gal-9在老年ACI患者中均呈高表达,可用于评估老年ACI患者超早期静脉溶栓治疗的预后,二者联合检测的评估价值更高。

柴胡疏肝散对胃肠功能紊乱大鼠结肠肌层缝隙连接蛋白43表达的影响

目的:探讨柴胡疏肝散对胃肠功能紊乱大鼠结肠肌层缝隙连接蛋白43(CX43)表达的影响。方法:将8周龄SD大鼠100只随机分为对照(sham)组、模型(FGID)组、FGID+药物组及sham+药物组,每组25只。对于FGID组、FGID+药物组,利用腹腔注射利血平的方式建立胃肠功能紊乱大鼠模型;sham组及sham+药物组大鼠实施生理盐水腹腔注射;完成造模后,为sham+药物组及FGID+药物组大鼠实施柴胡疏肝散汤剂灌服,sham组与FGID组大鼠灌服灭菌注射用水;通过实时荧光定量聚合酶链反应、Westeron Blot及组织免疫荧光技术检测四组大鼠结肠肌层CX43 mRNA、CX43蛋白表达水平及CX43阳性表达率。结果:四组大鼠结肠肌层CX43 mRNA、CX43蛋白表达水平及CX43阳性表达率比较,差异无统计学意义(P>0.05)。结论:柴胡疏肝散能够提高胃肠功能紊乱大鼠结肠肌层CX43的表达水平,从而改善胃肠功能。

外周血单个核细胞Cx37、AIM2、Cx43对冠心病的预测价值分析

目的探讨外周血单个核细胞缝隙连接蛋白37(Cx37)、黑色素瘤缺乏因子2(AIM2)、缝隙连接蛋白43(Cx43)对冠心病的预测价值。方法选取2019年1月至2023年10月该院收治的460例因“疑似冠心病或者胸闷、胸痛待查”而入院拟择期行冠状动脉造影(CAG)的患者作为研究对象,根据CAG检查结果分为冠心病组(212例)和非冠心病(248例)。比较两组临床特征及外周血单个核细胞Cx37、AIM2、Cx43,并予以多因素Logistic回归分析冠心病发生的危险因素,采用受试者工作特征(ROC)曲线分析外周血单个核细胞Cx37、AIM2、Cx43对冠心病的预测价值。结果冠心病组年龄、体质量指数大于非冠心病组(P<0.05),心率、血清糖化血红蛋白(HbA1c)、空腹胰岛素(FINS)水平高于非冠心病组(P<0.05),吸烟、高血压、高脂血症、冠心病家族史的患者占比高于非冠心病组(P<0.05),血清高密度脂蛋白胆固醇(HDL-C)水平低于非冠心病组(P<0.05)。冠心病组外周血单个核细胞Cx37、Cx43表达水平低于非冠心病组,外周血单个核细胞AIM2表达水平高于非冠心病组(P<0.05)。多因素Logistic回归分析结果显示,吸烟,高血压,高脂血症,冠心病家族史,外周血单个核细胞Cx37、Cx43表达水平降低,外周血单个核细胞AIM2表达水平升高是冠心病发生的独立危险因素(OR=2.568、2.375、3.313、2.713、0.647、0.464、2.995,P<0.05)。ROC曲线分析结果显示,外周血单个核细胞Cx37、AIM2、Cx43及3项联合检测预测冠心病的曲线下面积(AUC)分别为0.712、0.732、0.708、0.922,灵敏度分别为66.51%、70.28%、71.70%、89.15%,特异度分别为69.76%、70.16%、69.35%、82.26%,其中联合检测的AUC最大(P<0.05)。结论外周血单个核细胞Cx37、AIM2、Cx43对冠心病发生具有一定的预测价值,其中联合检测的预测价值最高。

WE43镁合金SLM成形数值模拟及试验验证

目的研究WE43镁合金激光选区熔化(SLM)成形过程、成形后变形及应力分布的变化规律,得到SLM态WE43常温拉伸力学模型。方法采用SLM方法制备了WE43镁合金悬臂梁及拉伸试样,通过对比悬臂梁局部切割翘曲试验结果与数值模拟结果,得到WE43镁合金固有应变模型,实现WE43镁合金SLM成形过程的模拟及变形预测;对SLM态WE43镁合金开展拉伸试验,使用金相显微镜及扫描电镜进行微观组织及断口形貌观察;采用Normalized Cockcroft&Latham模型对拉伸试验进行模拟,实现SLM态WE43常温拉伸过程分析。结论常温SLM态WE43的抗拉强度为313 MPa,屈服强度为236 MPa,延伸率为7.6%,试样中存在不规则孔洞缺陷;在SLM成形过程中,WE43镁合金固有应变值exx、eyy、ezz分别为−0.0025、−0.0025、−0.0115,悬臂梁最大翘曲高度为1.99 mm,模拟结果显示未切割悬臂梁最大等效应力为12.3 MPa;当NC&L断裂准则临界损伤值为0.1时,WE43常温拉伸过程模拟结果与试验结果最为接近,预测准确率为93%。

不同成形方式对WE43镁合金组织和力学性能的影响

采用激光选区熔化成形(SLM)、铸造及挤压方式制备了WE43镁合金试样,通过维氏硬度计、密度测试计、光学显微镜、扫描电镜以及拉伸试验机等设备分析了不同制备方式下WE43镁合金的宏微观组织和力学性能变化规律;设计了基于指数函数的模型对不同成形方式的WE43应力–应变曲线进行统一拟合,为WE43材料未来增材、减材以及等材工艺复合制造复杂零件打下基础。结果表明,SLM成形WE43有明显的各向异性,铸态和挤压态不明显。SLM成形WE43镁合金的强度最高,抗拉强度达到313 MPa,是铸态的183%;挤压态WE43镁合金塑性最好,伸长率达到10.2%,是铸态的232%;此外,SLM态镁合金密度只有1.731 g/cm^(3),仅为挤压态的85.7%和铸态的95.2%。在断裂特性上,SLM态和挤压态为韧性断裂,而铸造态为脆性断裂。在内部存在20μm左右孔洞形缺陷的情况下,SLM成形镁合金依然具有最高的强度,主要原因是SLM成形WE43镁合金平均晶粒尺寸仅为2.6μm,基体内存在大量的稀土相沉淀以及纳米级亚稳相。由此可知,通过进一步的后处理方法焊合SLM态镁合金内部孔洞形缺陷后,材料力学性能可以大幅提高。

缝隙连接蛋白43经典与非经典作用在疾病治疗中的潜在价值

背景:缝隙连接蛋白43在维持组织代谢稳态平衡中发挥着重要作用,传统观点认为它参与了缝隙连接过程,为细胞与细胞之间建立直接的物质信号交换通道奠定结构基础。而近年来研究对于其独特的半通道作用提出了新的看法,并发现了其亚细胞定位、自身片段等对于细胞生理活动及病理过程的重要意义。目的:综述数据库中相关文献,系统性总结缝隙连接蛋白43分子特征及在多种细胞表达的研究进展,重点阐述通道依赖性与非通道依赖性缝隙连接蛋白43的生理与病理作用,并探讨其在疾病治疗中的潜在价值。方法:分别设置“gap junction,connexin 43(Cx43),hemichannel,channel-dependent Cx43,channel-independent Cx43,extracellular vesicles(EVs),mitochondria,GJA1-20k”为英文关键词,以“缝隙连接,缝隙连接蛋白43,半通道,通道依赖性Cx43,非通道依赖性Cx43,线粒体,细胞外囊泡,GJA1-20k”为中文关键词,分别在PubMed数据库及中国知网数据库进行文献检索,最终共入选81篇文献进行综述分析。结果与结论:(1)缝隙连接蛋白43的经典作用即构成缝隙连接通道,通道依赖性缝隙连接蛋白43主要可通过直接构成缝隙连接通道参与组织器官的生理或病理过程,应充分关注其结构和功能的完整性,而黏附是缝隙连接的重要特性,与屏障障碍类疾病密切相关。(2)缝隙连接蛋白43的非经典作用即非缝隙连接通道依赖性作用,缝隙连接蛋白43六聚体目前被发现定位于质膜、线粒体内膜和细胞外囊泡表面等结构,参与炎性疾病的正向促炎机制、线粒体功能代谢和细胞外囊泡的靶向摄取等,选择性截短片段则参与全长连接蛋白43靶向转运至胞内各结构域过程,并且通过促使线粒体周围肌动蛋白聚合,调控线粒体稳态。(3)以上两种作用为开发靶向治疗药物及基于组织工程技术的治疗手段中种子细胞转化机制等问题的解决提供了新思路,但现存的一些原始研究常不能全面考虑不同形式缝隙连接蛋白43的相互作用,从而混杂地描述了其总体特征,使得研究结果产生偏差。(4)未来研究需要系统地构架不同形式存在的缝隙连接蛋白43的生理特性及在各疾病中的潜在机制,为缝隙连接蛋白43完整性机制探索及多疾病的诊断和治疗提供参考依据。

连接蛋白43半通道介导NLRP3炎症小体激活在脑缺血中的作用

缝隙连接蛋白在脑缺血后的神经炎症扩散中发挥重要作用。连接蛋白43(connexin 43,Cx43)作为中枢神经系统中主要的连接蛋白,通常以寡聚形式形成六聚体的半通道,与相邻细胞上的半通道对接,形成缝隙连接通道。在正常生理条件下,细胞表面的半通道开放维持在正常生理水平;然而,在脑缺血的过程中,Cx43半通道的过度开放导致了大量的离子(Na^(+)、Cl^(−)、Ca^(2+)、K^(+))、谷氨酸、天冬氨酸和三磷酸腺苷(ATP)等物质的释放,引起相邻细胞功能紊乱,从而加重神经细胞的损伤。此外,Cx43半通道的开放还诱导炎症因子的释放,这与脑缺血后NLRP3炎症小体的激活密切相关。因此,通过调控Cx43半通道能够缓解脑缺血后神经炎症,进而减轻脑缺血损伤。本文重点综述了Cx43半通道蛋白与NLRP3炎症小体激活的关系,以及其在脑缺血中的作用,旨在为脑缺血的治疗提供新的思路和方法。

Beclin-1联合Cx43对甲状腺癌的诊断价值及与甲状腺细针穿刺诊断准确率对比

目的 探讨自噬相关基因Beclin-1联合细胞缝隙连接蛋白43(Cx43)对甲状腺癌的诊断价值,并分析两者联合与甲状腺细针穿刺诊断对甲状腺癌准确率差异。方法 选取医院2020年10月至2023年10月收治的80例甲状腺癌患者作为观察组,另取同期收治的80例甲状腺结节患者作为对照组,取所有患者术后病理组织进行免疫组化染色,qRT-PCR仪进行检测与扩增,采用2-△△α法计算Beclin-1、Cx43的相对表达量,并将两组患者Beclin-1、Cx43水平进行对比。采用Spearman相关分析法分析Beclin-1、Cx43和甲状腺癌的相关性,并使用受试者工作特征(ROC)曲线分析Beclin-1联合Cx43诊断甲状腺癌的效果,并对比两者联合与甲状腺细针穿刺对甲状腺癌的诊断价值。结果 经免疫组化结果显示,两组细胞质内均检测到Beclin-1、Cx43,且观察组的Beclin-1、Cx43相对表达量比对照组更少(P<0.05),而Beclin-1、Cx43阳性率也同样小于对照组(P<0.05);Spearman相关分析结果显示,Beclin-1、Cx43和甲状腺癌呈现负相关(P<0.05);Beclin-1对甲状腺癌的诊断曲线下面积为0.822,最佳阈值为13.52;Cx43诊断甲状腺癌的曲线下面积为0.875,最佳阈值为18.67,两者联合的曲线下面积为0.932;Beclin-1联合Cx43对甲状腺癌的诊断灵敏度与特异度明显高于单一指标(P<0.05);Beclin-1联合Cx43对甲状腺癌的诊断准确率为93.75%,甲状腺细针穿刺对甲状腺癌的诊断准确率为98.75%,两者对比差异无统计学意义(P>0.05)。结论Beclin-1联合Cx43对甲状腺癌的诊断灵敏度与特异度较高,且两者联合与甲状腺细针穿刺诊断甲状腺癌准确率对比并无显著差异,可考虑将Beclin-1、Cx43两者作为甲状腺癌诊断及预后判断的重要标志物。