Chemokine platelet factor 4 accelerates peripheral nerve regeneration by regulating Schwann cell activation and axon elongation

Schwann cells in peripheral nerves react to traumatic nerve injury by attempting to grow and regenerate.Howeve r,it is unclear what factors play a role in this process.In this study,we searched a GEO database and found that expression of platelet factor 4 was markedly up-regulated after sciatic nerve injury.Platelet factor is an important molecule in cell apoptosis,diffe rentiation,survival,and proliferation.Further,polymerase chain reaction and immunohistochemical staining confirmed the change in platelet factor 4 in the sciatic nerve at different time points after injury.Enzyme-linked immunosorbent assay confirmed that platelet factor 4 was secreted by Schwann cells.We also found that silencing platelet factor 4 decreased the proliferation and migration of primary cultured Schwann cells,while exogenously applied platelet factor 4 stimulated Schwann cell prolife ration and migration and neuronal axon growth.Furthermore,knocking out platelet factor 4 inhibited the prolife ration of Schwann cells in injured rat sciatic nerve.These findings suggest that Schwann cell-secreted platelet factor 4 may facilitate peripheral nerve repair and regeneration by regulating Schwann cell activation and axon growth.Thus,platelet factor 4 may be a potential therapeutic target for traumatic peripheral nerve injury.

Platelet factor 4 induces bone loss by inhibiting the integrinα5-FAK-ERK pathway

Background:The effect of platelet factor 4(PF4)on bone marrow mesenchymal stem cells(BMMSCs)and osteoporosis is poorly understood.Therefore,this study aimed to evaluate the effects of PF4-triggered bone destruction in mice and determine the underlying mechanism.Methods:First,in vitro cell proliferation and cell cycle of BMMSCs were assessed using a CCK8 assay and flow cytometry,respectively.Osteogenic differentiation was confirmed using staining and quantification of alkaline phosphatase and Alizarin Red S.Next,an osteoporotic mouse model was established by performing bilateral ovariectomy(OVX).Furthermore,the PF4 concentrations were obtained using enzymelinked immunosorbent assay.The bone microarchitecture of the femur was evaluated using microCT and histological analyses.Finally,the key regulators of osteogenesis and pathways were investigated using quantitative real-time polymerase chain reaction and Western blotting.Results:Human PF4 widely and moderately decreased the cell proliferation and osteogenic differentiation ability of BMMSCs.Furthermore,the levels of PF4 in the serum and bone marrow were generally increased,whereas bone microarchitecture deteriorated due to OVX.Moreover,in vivo mouse PF4 supplementation triggered bone deterioration of the femur.In addition,several key regulators of osteogenesis were downregulated,and the integrinα5-focal adhesion kinase-extracellular signalregulated kinase(ITGA5-FAK-ERK)pathway was inhibited due to PF4 supplementation.Conclusions:PF4 may be attributed to OVX-i nduced bone loss triggered by the suppression of bone formation in vivo and alleviate BMMSC osteogenic differentiation by inhibiting the ITGA5-FAK-ERK pathway.

Clinical Evaluation of Platelet Rich Growth Factors (PRGFS) in Treatment of Temporomandibular Joint Disc Displacement—Study Report

Objective: The aim of the study was to evaluate the effect of platelet rich growth factors (PRGFs) in treatment of temporomandibular joint disc displacement. Materials and Methods: The study subjects included 8 females having bilateral anterior disc displacement with reduction and 1 female having bilateral anterior disc displacement without reduction with the age range between 20 - 35 years. The process of obtaining PRGFs was carried out following the Anitua Technique. Results: Clinical parameters of Interincisal distance, Lateral excursion of mandible using digital caliper in millimeters and Visual Analogue Scale (VAS 0/10) for pain intensity score were used. All of these parameters were running through the intervals of two, four, and eight weeks till the end of the follow-up period at twenty-six weeks (six months). The participated patients showed the clinical improvement in the different clinical statuses such as interincisal distance;lateral excursion of mandible and Pain Score. Conclusion: the study reported early efficacy of PRGFs after the arthrocentesis of the joint in treatment of TMJ disc displacement, and according to our results, the injection of PRGFs could be a possible alternative treatment for patients who did not respond to standard treatment.

Elevated Platelet Activating Factor Level in Ischemia-Related Arrhythmia and Its Electrophysiological Effect on Myocardium

Objective The mechanism through which platelet activating factor （PAF） induces cardiac electrical activity and arrhythmia is not well understood and previous studies have suggested a potential involvement of ion channels in its action. The present study was aimed to clarify the role of PAF in fatal arrhythmias following acute myocardia infarction （AMI） and the underlying mechanism. Methods （1） Blood PAF levels were measured among 72 AMI patients at the time of diagnosis with AMI and 48 h later, and their electrocardiogram （ECG） was recorded continuously. （2） Ischemia simulation and surface electrocardiogram were conducted in 20 pigs and their PAF levels were measured. （3） PAF perfusion and standard microelectrode recording were performed on guinea pig papillarymuscles. Results In both humans and pigs, elevated PAF levels were detected in AMI and simulated ischemia, respectively, and even higher PAF levels were found when fatal arrhythmias occurred. In guinea pig myocardium, PAF induced a shortening of action potential duration at 90% level of repolarization （APD 90 ）under non-ischemic conditions and a more pronounced shortening under early simulated ischemic conditions. Conclusion AMI and ischemia are associated with increased PAF levels in humans and pigs, which are further raised when fatal arrhythmia follows. The effects of PAF on the myocardium may be mediated by multiple ion channels.

Sequential expression of cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor in rat hippocampal neurons after fluid percussion injury

Traumatic brain injury causes gene expression changes in different brain regions. Occurrence and development of traumatic brain injury are closely related, involving expression of three factors, namely cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor. However, little is known about the correlation of these three factors and brain neuronal injury. In this study, primary cultured rat hippocampal neurons were subjected to fluid percussion injury according to Scott’s method, with some modifications. RT-PCR and semi-quantitative immunocytochemical staining was used to measure the expression levels of cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor. Our results found that cyclooxygenase-2 expression were firstly increased post-injury, and then decreased. Both mRNA and protein expression levels reached peaks at 8 and 12 hours post-injury, respectively. Similar sequential changes in glutamate receptor 2 were observed, with highest levels mRNA and protein expression at 8 and 12 hours post-injury respectively. On the contrary, the expressions of platelet activating factor receptor were firstly decreased post-injury, and then increased. Both mRNA and protein expression levels reached the lowest levels at 8 and 12 hours post-injury, respectively. Totally, our findings suggest that these three factors are involved in occurrence and development of hippocampal neuronal injury.

Prolonged propagation of rat neural stem cells relies on inhibiting autocrine/paracrine bone morphogenetic protein and platelet derived growth factor signals

Continuous expansion of rat neural stem cell lines has not been achieved due to proliferation arrest and spontaneous differentiation in vitro. In the current study, neural precursor cells derived from the subventricular zone of adult rats spontaneously underwent astroglial and oligodendroglial differentiation after limited propagation. This differentiation was largely induced by autocrine or paracrine bone morphogenetic protein and platelet derived growth factor signals. The results showed that, by inhibiting bone morphogenetic protein and platelet derived growth factor signals, adult rat neural precursor cells could be extensively cultured in vitro as tripotent stem cell lines. In addition to adult rat neural stem cells, we found that bone morphogenetic protein antagonists can promote the proliferation of human neural stem cells. Therefore, the present findings illustrated the role of autocrine or paracrine bone morphogenetic protein and platelet derived growth factor signaling in determining neural stem cell self-renewal and differentiation. By antagonizing both signals, the long-term propagation of rat neural stem cell lines can be achieved.

Effects of Platelet Factor 4 on Expression of Bone Marrow Heparan Sulfate in Syngenic Bone Marrow Transplantation Mice

To explore the effects of platelet factor 4(PF4) on hematopoietic reconstitution and its mechanism in syngenic bone marrow transplantation (BMT). The syngenic B MT mice models were established. 20 and 26 h before irradiation, the mice were injected 20 μg/kg PF4 or PBS twice into abdominal cavity, then the donor bone marrow nuclear cells (BMNC) were transplanted. On the 7th day, spleen clone forming units (CFU S) were counted. On the 7th, 14th and 21st day after BMT, the BMNC and megakaryoryocytes in bone marrow tissue were counted and the percentage of hematopoietic tissue and expression level of heparan sulfate in bone marrow tissue were assessed. In PF4 treated groups, the CFU S counts on the 7th day were higher than those in BMT groups after BMT. The BMNC and megakaryoryocyte counts and the percentage of hematopoietic tissue and heparan sulfate expression level were higher than those in BMT group on the 7th, 14th and 21st day after BMT ( P <0.01 or P <0.05). PF4 could accelerate hematopoietic reconstitution of syngenic bone marrow transplantation. The promotion of the heparan sulfate expression in bone marrow may be one of mechanisms of PF4.

In vitro effects of buyang huanwu decoction and its ingredients on inhibiting the specific binding of ~3H-platelet activating factor to its receptor in rabbits

BACKGROUND： Pharmacologic action of traditional Chinese medicine compound is the comprehensive effect of various ingredients, and the interactions of various ingredients are closely correlated with the final effect. In order to reveal the compatibility mechanism of buyang huanwu decoction （BHD）＇s prescription in treating and preventing ischemic cerebrovascular disease, we need to explore the effect and relation of ingredients in prescription except for considering the effect of each ingredient on the whole prescription. OBJECTIVE： To study the effect of BHD and its ingredients in the prescription on the specific binding of 3H-platelet activating factor （PAF） to its receptor （PAFR）in rabbits in vitro, and to analyze the action of each ingredient in the prescription. DESIGN： A decomposed recipe study based on orthogonal test. SETTING： Guangzhou University of Traditional Chinese Medicine. MATERIALS： Five healthy adult New Zealand rabbits of either gender were provided by the Experimental Animal Center of Guangzhou University of Traditional Chinese medicine. The prescription herbal pieces were purchased from Foshan Kangpu Pharmaceuticals Company and Jianmin Pharmaceuticals Company, and were appraised by Professor Yanchen Xu from College of Traditional Chinese Medicine, Guangzhou University of Traditional Chinese Medicine. 3H-PAF was supplied by Amersham Co,Ltd.（Specific activity： 6.475 TBq/mmol;batch number：200402）; PAF standard by Biomol Co., Ltd.（batch number： P1318V）. METHODS： This experiment was carried out in the Laboratory of Nuclear Medicine, Guangzhou University of Traditional Chinese Medicine between September and December 2004. ①The seven influencing factors were selected： such as Shenghuangqi , Dangguiwei, Chishao, Dilong, Taoren, Honghua, Chuanxiong. Each factor was divided into two levels, selected or not selected. The tests were arranged according to L8 （27） orthogonal test table. ②The specific binding of 3H-PAF to its receptors in rabbits was measured by radioligand binding assay. The inhibitory rate of the specific binding was used as an assessing index. The inhibitory action of and on 3H-PAF to PAFR binding was analyzed and compared in vitro. The inhibitory action of each ingredient in the prescription BHD on 3H-PAF to PAFR binding was investigated and compared in vitro by direct analysis and analysis of variance of orthogonal test. MAIN OUTCOME MEASURES： Effect of 8 prescriptions for L8 （27） orthogonal test table on the specific binding inhibition rate of 3H-PAF and PAFR. RESULTS： According to results of variance analysis of orthogonal test, the inhibitory action of each ingredient in the prescription BHD on 3H-PAF to PAFR binding from the highest to the lowest was in turn Honghua, ShenghuangqL Taoren, Dilong, DangguiweL Chuanxiong, Chishao. Honghua, Shenghuangqi, Taoren, Dilong, Danguiwei were major influence factors to 3H-PAF to PAFR in rabbits （F = 187.829,144.446,59.521,5.018,4.265, P 〈 0.05- 0.01）, but Chuanxiong and Chishao had not obviously inhibitory effect. The specific binding inhibition rate of prescriptions （except Shenghuangqi ） was obviously higher than that of one of prescriptions （Shenghuangqi included）. CONCLUSION： The results of orthogonal test show that Honghua, ShenghuangqL Taoren, Dilong, Dangguiwei are major influencing factors to inhibit binding of sH-PAF to PAFR in rabbits, among which, Honghua is the strongest in ingredients of prescription BHD. The results also reveal that Shenghuangqi is able to weaken the inhibitory effect and to prevent the strong inhibitory effect of blood-activating drugs in BHD.

Effects of Buyang Huanwu decoction and Astragalus mongholicus on platelet activating factor receptor activity in rabbits in vitro

BACKGROUND: The pharmacological action of traditional Chinese medicine compound is the comprehensive effect of the various ingredients, and the interactions of various ingredients are closely correlated with the final effect. In order to reveal the compatibility mechanism of BHD's prescription in treating and preventing ischemic cerebrovascular disease, we needed explore the effect and relation of ingredients in the prescription. OBJECTIVE: To observe the effect of Buyang Huanwu decoction (BHD) and Astragalus mongholicus on the activity of platelet activating factor receptor (PAFR) in the platelet of rabbits in vitro, and investigate the mechanism of Astragalus mongholicus. DESIGN: A decomposed recipes study. SETTING: Guangzhou University of Traditional Chinese Medicine. MATERIALS: Five New Zealand rabbits, weighing 2-3 kg, both sexes, were used. BHD was composed of Sheng Huang Qi 120 g, Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g, Hong Hua 3 g. The prescription for activating blood circulation consisted of Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g and Hong Hua 3 g. The prescription for invigorating qi consisted of 120 g Sheng Huang Qi. The prepared herbal pieces were purchased from the traditional Chinese medicine Dispensary of Foshan Second People's Hospital, and appraised by Professor Xu from Science of Chinese Materia Medica College, Guangzhou University of Traditional Chinese Medicine. 3H-PAF was supplied by Amersham Co., Ltd. (specific activity: 6. 475 TBq/mmol; batch number: 200402); PAF standard by Biomol Co., Ltd. (batch number: P1318V). METHODS: The experiments were carried out in the Laboratory of Nuclear Medicine, Guangzhou University of Traditional Chinese Medicine from September to December 2004. ① Injections of BHD, prescriptions for activating blood circulation and invigorating qi were prepared by the decoction and alcohol sedimentation technique. Rabbit common carotid artery blood (40 mL) was drawn via intubation to prepare platelet suspension of (0.8-1.0)×1010 L-1. ② Determination of 3H-PAF and washed PAFR binding: The general combination tube (T) contained washed platelet-rich plasma (WPRP) 380 μL + 3H-PAF (0.35 nmol/L)10 μL+distilled water 5 μL; The nonspecific binding tube (P) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+cold PAF (1 μmol/L) 5 μL; The sample tube (Y) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+experimental medicine (injection of BHD, prescriptions for activating blood circulation or invigorating qi) 5 μL. The test was conducted for three times for each sample in the same way as mentioned above. The samples were shaken on the oscillator for 30 s, then bathed at 25 ℃ for 40 minutes, and the reaction was terminated with cold Tris buffer containing 0.1% BSA, multichannel cell detachment separator was used for vacuum suction to filter the separated free 3H-PAF, and the filter paper was washed with cold Tris buffer for four times, then dried in the baking oven (80 ℃) for 1 hour, and placed in xylol liquid scintillator, and the radioactivity was determined automatically by the liquid scintillation detector. The mean of the three parallel tubes was calculated. The specific binging inhibition rate was calculated: SBIR=[(T-Y)/(T-P)]×100%]. ③ Univariate analysis of variance was conducted. And for comparison of each paired groups, the q test was adopted. MAIN OUTCOME MEASURES: Effect of BHD whole prescription, prescriptions for activating blood circulation and invigorating qi on the specific binding inhibition rate of 3H-PAF and PAFR. RESULTS: BHD, prescriptions for activating blood circulation and invigorating qi were all able to inhibit the specific binding of 3H-PAF to PAFR, the specific blinding inhibition rates were (45.90±7.50)%, (97.90±1.84)% and (26.75±2.48)%, respectively, and there were significant differences between every two groups (P < 0.01). CONCLUSION: Single Astragalus mongholicus (120 g) can inhibit the specific blinding of PAFR in the platelet of the rabbit with 3H-PAF, but the combination of Astragalus mongholicus with the drugs for activating blood circulation in BHD can significantly decrease the inhibiting action of the latter on PAFR activity of the platelet, reflecting the combined mechanism of 'removing blood stasis without injuring the vital qi' in BHD.

Up and Down Expression of Androgen Receptor, Estrogen Receptor beta and Platelet Derived Growth Factor beta by Testosterone in Aortic Vascular Smooth Muscle Tissues

Objectives To investigate the effects of testosterone enanthate (TE) on serum lip- ids and lipoproteins metabolism and the expression of androgen receptor (AR) , estrogen receptor beta ( ER - β) and platelet derived growth factor beta ( PDGFR - β) in aortic vascular smooth muscle tissues (VSMTs). Methods Forty aged male rats were ran- domly divided into 4 groups, group A (placebo group) , group B (2. 5 mg/kg intramuscular injection of TE once a week ) , group C (5.0 mg/kg intramuscular injection of TE once a week ) , group D ( 10. 0 mg/kg intramus- cular injection of TE once a week). All animals were fed freely during 16 - week treatment periods. The ex- pression of AR ,ER - βand PDGFR - β were studied by Western bolt. Results Average serum LDL - C was lower in group D than that in group A ( p < 0. 01 ). Compared with the other groups, average serum TC was also lower in group D (p <0. 05). AR expression in aortic vascular smooth muscle tissues could be regulated by TE: 99.50 ±21.74, 125.38 ±28.68 and 101.98 ± 15.42 for TE concentrations at 2.5 mg/kg, 5.0 mg/kg and 10.0 mg/kg, respectively , the expression of ER - β could be regulated by TE: 92. 34 ± 18. 68, 47. 72 ± 18.12, 82.13 ±23.50, and the expression of PDGFR - β could be regulated as well by TE: 219.70 ±45. 59, 50.16 ±9. 72, 125.36 ±15. 74(Data for AR ,ER-β and PDGFR - β protein band intensity were expressed with x ± s, with control group taken as 100 ).Conclusions This study indicates that androgens have significant effects on serum lipids and lipoprotein metabolism. Testosterone enanthate at 5. 0 mg/kg can stimulate the expression of AR, but inhibite the expres- sion of PDGFR. Testosterone enanthate at the concen- trations of 5. 0 mg/kg and 10. 0 mg/kg can inhibite the expression of ER - β.

非酒精性脂肪性肝病患者C/RL-r、APRI、FIB-4水平与肝纤维化发生的相关性

目的分析非酒精性脂肪性肝病(NAFLD)患者改良肝尾状叶/右叶比值(C/RL-r)、天门冬氨酸氨基转移酶与血小板比值指数(APRI)、基于4因子的纤维化指数(FIB-4)与肝纤维化发生的相关性。方法选择2021年2月至2022年12月在保定市第一中心医院治疗的NAFLD患者153例,根据病理学结果,将患者分为无肝纤维化组81例、肝纤维化组72例。行MRI扫描检测C/RL-r;计算APRI、FIB-4水平;分析NAFLD患者实验室指标、C/RL-r、APRI、FIB-4水平与发生肝纤维化的相关性,发生肝纤维化的独立危险因素及C/RL-r、APRI、FIB-4对NAFLD患者发生肝纤维化的预测价值。结果肝纤维化组ALT、AST、TBil、GGT、TG、C/RL-r、APRI、FIB-4水平显著高于无肝纤维化组,分别为(42.32±10.21)U/L比(36.21±7.78)U/L、(45.36±8.72)U/L比(27.45±5.40)U/L、(13.52±3.65)μmol/L比(12.24±2.16)μmol/L、(60.53±13.41)U/L比(53.69±12.44)U/L、(1.99±0.53)mmol/L比(1.05±0.33)mmol/L、(1.15±0.12)比(0.92±0.09)、(0.52±0.15)比(0.32±0.10)、(1.47±0.47)比(0.94±0.30),高密度脂蛋白胆固醇(HDL-C)水平显著低于无肝纤维化组为(1.03±0.26)mmol/L比(1.32±0.45)mmol/L,(t=4.189、15.453、2.674、3.272、13.322、13.501、4.302、8.405、4.801,均P<0.05);NAFLD患者ALT、AST、TG、C/RL-r、APRI、FIB-4与发生肝纤维化呈正相关(r=0.531、0.435、0.571、0.605、0.771、0.716,均P<0.001);ALT、AST、TG、C/RL-r、APRI、FIB-4水平高是影响NAFLD患者发生肝纤维化的独立危险因素(P<0.05);C/RL-r、APRI、FIB-4、三者联合预测NAFLD患者发生肝纤维化的曲线下面积(AUC)分别为0.767、0.830、0.754、0.936;相较于C/RL-r、APRI、FIB-4单独预测的AUC,三者联合预测的AUC更高(Z=4.495、3.999、4.677,均P<0.001)。结论发生肝纤维化的NAFLD患者C/RL-r、APRI、FIB-4水平较高,三者联合检测对NAFLD患者发生肝纤维化具有较高预测价值。

非侵入性肝纤维化评分在肝硬化并食管胃底静脉曲张严重程度预测中的运用

目的 评估无创检查预测食管静脉曲张严重程度及出血的可能性。方法 收集2021年1月至2022年5月包头医学院第二附属医院86例肝病患者的临床资料,包括实验室检查、影像学检查、胃镜检查。根据胃镜检查评估食管静脉曲张严重程度。APRI、GPR、FIB-4、S-index、AAR、GAPRI对肝硬化伴食管静脉曲张出血严重程度的预测价值。结果 APRI、GPR、FIB-4、S-index、AAR、GAPRI评估肝硬化食管胃底静脉曲张出血严重程度的曲线下面积分别为0.646、0.637、0.634、0.632、0.566、0.636,特异度分别56.72%、89.74%、36.84%、73.68%,57.89%、89.47%,敏感度分别为56.72%、40.30%、91.04%、55.22%、61.19%、40.30%。食管静脉曲张轻度组中血小板、ALT、RDW分别为(114.47±50.71)×109、(34.74±15.81)U/L、(16.889±2.252),严重组为(95.79±42.27)×10^(9)、(42.52±24.97)U/L、(15.849±2.602),差异均无统计学意义(P>0.05)。轻度组APRI、FIB-4、S-index、 GAPRI、 GPR、AAR分别为0.637(0.409,0.895)、0.637(0.409,0.895)、3.562(2.166,6.155)、0.435(0.211,0.984)、43.810(26.829,64.021)、0.730(0.447,1.067)、1.555±0.623,严重组分别为0.901(0.524,1.407)、0.901(0.524,1.407)、4.5(3.335,6.991)、0.891(0.327,1.778) 61.000(29.126,144.444)、1.017(0.485,2.407)、1.482±0.720,差异无统计学意义(P>0.05)。结论 APRI、GPR、GAPRI、S-index对肝硬化伴食管静脉曲张出血严重程度有一定预测价值,其中APRI预测价值更大。

PF4对造血干/祖细胞系KG1a总粘附性、粘附分子表达及细胞骨架蛋白聚合的作用

目的研究ＰＦ４单用及与ＩＬ－３联合应用对造血干／祖细胞系ＫＧ１ａ总粘附性、粘附分子表达及细胞骨架蛋白聚合的影响。方法采用结晶紫染色、免疫标记流式细胞仪测定、ＭＴＴ、荧光分光光度等方法。结果１．单用１００ｎｇ／ｍｌ ＰＦ４作用于ＫＧ１ａ细胞时，ＫＧ１ａ细胞总粘附性增长８０％，单用２０ｎｇ／ｍｌ ＩＬ－３作用于ＫＧ１ａ细胞时，ＫＧ１ａ细胞总粘附性增长９６％，ＰＦ４与ＩＬ－３联合作用于ＫＧ１ａ细胞时，ＫＧ１ａ细胞总粘附性增长９７％。２．ＰＦ４作用于ＫＧ１ａ后，ＣＤ３１、ＣＤ４４、ＣＤ１１ａ、ＣＤ１１ｂ表达分别增加了２１％、２２％、１７％、１８％，较前均有显著性（Ｐ＜０．０５＝，而ＰＦ４对ＫＧ１ａ细胞的ＣＤ６２Ｐ、ＣＤ６２Ｅ表达无影响；ＰＦ４与ＩＬ－３联合作用于ＫＧ１ａ时，ＣＤ３１、ＣＤ４４、ＣＤ１１ａ、ＣＤ１１ｂ表达并没出现互相促进作用。３．分别用抗ＣＤ３１、ＣＤ４４、ＣＤ１１ａ、ＣＤ１１ｂ等单克隆抗体阻断ＫＧ１ａ粘附分子时，对ＰＦ４引起的ＫＧ１ａ粘附性分别下调了３９％、４３％、３４％、３８％。４．ＰＦ４、ＩＬ－３作用ＫＧ１ａ细胞时，能够引起ＫＧ１ａ细胞肌动蛋白的聚合。结论ＰＦ４可刺激早期的白血病性造血干／祖?

血小板第四因子PF4对人中性粒细胞粘附性与呼吸爆发的调节作用

目的研究血小板第四因子PF4对人中性粒细胞Np功能的影响。方法用结晶紫染色、免疫荧光标记、PKC试剂盒测定、NBT法及高香草酸HVA荧光分光光度法比较对照组与PF4作用的实验组在趋化因子刺激前后,PF4对中性粒细胞总粘附性、整合蛋白CD11b的表达、PKC水平及呼吸爆发作用产生活性氧物质reactiveoxygenspeciesROSO2·-、H2O2等的影响。结果PF4对静息中性粒细胞总粘附性有轻度的增强作用使总粘附性上调了12.24%P<0.01,而对整合蛋白CD11b的表达、呼吸爆发产生的ROSO2·-、H2O2则无影响。进一步研究发现PF4使经趋化三肽(FMLP)活化的中性粒细胞的总粘附性下调17.47%P<0.01;使经趋化三肽(FMLP)活化的中性粒细胞的整合蛋白CD11b的表达下调18.84%P<0.01;使经佛波酯(PMA)活化的中性粒细胞的呼吸爆发作用产生的O2·-、H2O2分别下调了50.29%P<0.01、226.53%P<0.05。研究同时发现PF4对静息及活化的中性粒细胞的PKC水平均无影响。结论首次证明PF4与其它经典的趋化因子不同,是中性粒细胞活化的一个负调节因子,对中性粒细胞功能主要起降调节的作用。这种降调节作用不是通过PKC信号转导途径完成的。

血小板第四因子对脐血CD34^+细胞趋化作用的研究

目的 :研究PF4及其小肽PF417 70对新鲜脐血CD34+细胞的趋化作用及对粘附分子表达的影响。方法 :采用免疫磁珠法 (MACS)分选CD34+细胞 ,利用Transwell穿孔板测定PF4对CD34+细胞的趋化作用 ;流式细胞仪检测免疫荧光标记的粘附分子及CXCR4的表达。结果 :①PF4对脐血CD34+细胞有趋化作用 ,PF4组的趋化百分比为 15 7.43%± 5 0 .0 6 %(P <0 .0 5 ) ,PF417 70组为 187.0 2 %± 10 .6 9%(P <0 .0 5 )。②PF4作用于CD34+细胞时 ,CD49d和CXCR 4表达增加 ,对其它粘附分子CD31,CD44 ,CD11a ,CD6 2 p ,CD6 2E的表达没有影响。 结论 :PF4对脐血CD34+细胞有趋化作用 ,促进整合素CD49d及CXCR4的表达 ,PF4有助于脐血干细胞的归巢。

血小板第四因子在原核中的高效表达、分离纯化及活性测定

构建重组表达质粒pET - 32c PF4 ,并在原核中进行表达 ,以探讨其对白血病细胞系 (HEL)细胞增殖的作用。方法 :通过PCR方法从含有PF4基因的PQE - 6 0 PF4质粒中扩增PF4 ,用NcoⅠ HindⅢ双酶切 ,克隆到原核表达质粒pET - 32C中 ,使之在BL2 1表达 ,Ni-ChelatingSepharose亲和柱纯化 ,用细胞集落形成方法研究重组PF4对HEL的抑制作用。结果 :菌株筛选后在大肠杆菌中获得可溶性高效表达 ,重组PF4占菌体总蛋白的 2 2 %。肠激酶酶切除去N -端融合部分 ,获得了高纯度的重组人血小板第四因子 (rh -PF4 ) ,纯度为 95%以上。活性实验发现重组PF4可抑制HEL细胞集落的形成 ,抑制率为 4 7%。结论 :原核表达质粒pET - 32C可高效可溶性表达PF4 ,重组PF4对HEL细胞的增殖有抑制作用。

APRI、FIB-4及Fibro Touch联合检测对乙肝肝纤维化的早期预警价值

目的探讨APRI、FIB-4及Fibro Touch联合检测对乙肝肝纤维化的早期预警价值。方法回顾性分析确诊为慢性乙型肝炎的患者402例,根据肝活组织检查结果分组:无明显肝纤维化组(S_0-S_1)146例,明显肝纤维化组(S_2-S_3)162例、早期肝硬化组(S_4)94例。每例患者进行APRI、FIB-4及Fibro Touch的评估。计算上述无创指标联合检测与肝组织病理的相关性,分析无创诊断模型对乙肝患者肝纤维化(≥S_2)的早期诊断价值,同时绘制受试者工作特征(ROC)曲线,计算最佳预测值、灵敏度及特异性。结果 APRI、FIB-4及Fibro Touch 3种无创诊断方法与肝活组织检查具有良好一致性(P<0.05),其Spearman相关系数分别为0.768、0.712、0.865。根据肝组织病理结果将入组病例分为无明显肝纤维化组(S_0-S_1)与肝纤维化组(≥S_2),对无创诊断模型行多因素二元Logistic回归分析提示APRI、FIB-4及Fibro Touch对肝纤维化均有一定的预测价值(P<0.05),其OR值分别为1.996、1.563及2.180。以APRI、FIB-4、Fibro Touch作为参数拟合Logistic二元回归方程,拟合优度高(χ~2=13.689,P=0.126),得出上述指标联合检测对肝纤维化早期预警方程为logit(P)=-0.556+1.119*(APRI)+1.202*(FIB-4)+1.682*(Fibro Touch)。ROC曲线分析显示3种方法联合检测对于肝纤维化(≥S_2)的早期预警价值最大,Fibro Touch检测预警效能优于APRI、FIB-4,而APRI和FIB-4在肝纤维化的早期预警价值无显著差异。采用上述指标联合对于肝纤维化(≥S_2)的早期预警曲线下面积为0.928,其最大约登指数为0.819,最佳临界值为18.14,对应的灵敏度、特异度分别为89.6%和92.3%。结论 APRI、FIB-4及Fibro Touch联合检测对乙肝肝纤维化具有较高的早期预警价值,值得推广。

Fibroscan、APRI及FIB-4用于HBV相关肝硬化的诊断价值研究

目的探讨Fibroscan、APRI及FIB-4用于乙型肝炎病毒(HBV)相关肝硬化诊断的诊断价值。方法纳入2013年3月至2016年3月就诊的HBV相关肝硬化患者129例(肝硬化组),同时设置慢性乙型肝炎患者116例作为对照(CHB组)。患者就诊时收集一般资料及常规实验室指标,根据公式计算APRI、FIB-4值。固定1名医生对患者进行Fibroscan检测,记录肝硬度值(LSM)。分析临床指标之间的相关性,并使用ROC曲线检测LSM、APRI及FIB-4用于区别诊断CHB与HBV相关肝硬化的价值。结果肝硬化组患者LSM、APRI及FBI-4显著高于CHB组,差异有统计学意义(P<0.05)。肝硬化组中,LSM与ALT、AST、APRI、FIB-4呈正相关(r值分别为0.241、0.283、0.417、0.485,P<0.05),与PLT呈负相关(r=-0.305,P<0.05);CHB组中LSM与各项指标均无相关性(P>0.05)。LSM、APRI及FIB-4诊断肝硬化的AUC分别为0.831(95%CI:0.780~0.881)、0.717(95%CI:0.651~0.782)、0.715(95%CI:0.650~0.780)。结论 Fibroscan、APRI及FIB-4均可用于区别诊断低炎症活动CHB及HBV相关肝硬化,且Fibroscan诊断效能优于APRI及FIB-4。

Effects of Xinfeng Capsule(新风胶囊) on the Expression of Platelet Derived Growth Factor in Synovium of Adjuvant Arthritis Rats

Objective： To observe the effects of Xinfeng Capsule （新风胶囊, XFC） on platelet parameters in peripheral blood and expression of platelet derived growth factor （PDGF） in synovium of adjuvant arthritis （AA） rats. Methods： A total of 40 male Sprague-Dawley （SD） rats were randomized into 5 groups： normal control （NC）, AA model control （MC）, methotrexate （MTX） treatment, Tripterygium wilfordii polycoride tablet （TPT） treatment, and XFC treatment. Excluding the NC group, the AA model was induced by intracutaneous injection of 0.1 mL Freund＇s complete adjuvant in the right hind limb. Induction of AA and the effects of drug treatments were assessed by voix pedis swelling, arthritis index （AL） body mass, and the pathological changes of joints and cartilage with a light microscopy. Platelet parameters in peripheral blood were detected with an automated hematology analyzer. PDGF in synovium was detected with immunohistochemical methods and PDGF mRNA expression in synovium was detected with reverse transcription polymerase chain reaction. Results： Compared with the NC group, the MC group had significantly increased voix pedis swelling, AI, platelet （PLT） and plateletcrit （PCT） in peripheral blood and PDGF as well as PDGF mRNA in synovium （all P〈0.01） and the joint cartilage was also highly degenerated. Compared with the MC group, the 3 treated groups had significantly decreased voix pedis swelling, AI, PLT, PCT, PDGF, and PDGF mRNA （P〈0.01）. The body mass in the XFC group was significantly higher than those in MTX and TPT groups （P〈0.05）. The levels of PLT, PCT, PDGF, and PDGF mRNA in the XFC group showed a decreasing tendency with no significant difference compared with the M＇IX and TPT groups （P〉0,05）. PDGF and PDGF mRNA of AA rats were positively correlated with voix pedis swelling, AI, PLT, and PCT （P〈0.05 or P〈0.01）. Conclusions： The expression and biosynthesis of PDGF increase in the synovium of AA rats and correlate with voix pedis swelling, AI, PLT, and PCT. XFC can decrease the levels of PDGF, PDGF mRNA, PLT, and PCT, thereby mitigating inflammation induced by platelet activation and reducing voix pedis swelling and the AI in AA rats.