【摘要】 目的 探讨机采血小板在实验室内保存期间的生化改变及其意义。 方法 采用ROCHE cobas b 221和OLYMPUS AU400测定血小板样本的pH值、PO2、PCO2、葡萄糖（GLU）浓度、乳酸脱氢酶（LDH）释放量。 结果 保存期间pH值由7.311（第1天）降至7.116（第5天）；PO2波动于106.7～119.5 mm Hg（1 mm Hg=0.133 kPa）；PCO2波动于25.0～35.8 mm Hg；GLU浓度呈进行性下降，由18.8 mmol/L降至15.2 mmol/L；LDH浓度显著增加，由150 mmol/L（第1天）增至259 mmol/L（第5天）。 结论 血小板在保存期内，其生化参数会随着保存时间发生改变。而这些参数作为间接指标对机采血小板是否被细菌污染具有一定的指导意义。【Abstract】 Objective To observe the biochemistry changes of apheresis platelets (AP), including pH, PO2, PCO2, GLU, LDH, within the storage time and its significance. Methods The pH, PO2, PCO2, GLU and LDH of platelet samples were detected by ROCHE cobas b 221 and OLYMPUS AU400. Results During the storage time, the pH value of platelet decreased gradually from 7.311 (the first day) to 7.116 (the fifth day); the PO2 fluctuated from 106.7 mm Hg (1 mm Hg=0.133 kPa) to 119.5 mm Hg; the PCO2 fluctuated from 25.0 mm Hg to 35.8 mm Hg;the concentration of GLU decreased from 18.8 mmol/L to 15.2 mmol/L;the concentration of LDH increased observably from 150 mmol/L (the first day) to 259 mmol/L (the fifth day). Conclusion There were observably changes of biochemistry of the AP during storage time, and these changes may be the indicators of platelets contamination.
OBJECTIVE: To study the effect of platelet-rich plasma in the repair of bone defect. METHODS: Segmental bone defects of 1 cm were created in the mid-upper part of bilateral radius of 24 New Zealand white rabbits. One side was randomly chosen as the experimental side, which was filled with artificial bone with platelet-rich plasma (PRP). The other side filled with artificial bone without PRP as the control. After 2, 4, 8 and 12 weeks of implantation, the gross, radiological, histological observations, and computer graphic analysis were performed to investigate the bone healing of the defect in both sides. RESULTS: Two weeks after operation, new bone and fibrous tissue formation in both the experimental and the control sides were observed only in the areas adjacent to the cut ends of the host bone, but the amount of new tissue in the experimental side was much more than that in the control side. In the 4th and 8th weeks, the surface of the artificial bone was covered with a large amount of new bones, the artificial bone was bridged tightly with the host bone by callus in the experimental side, while new bone was limited mainly in the cut ends and was less mature in the control side. In the 12th weeks, bone defects were entirely healed in the experimental side, which were covered completely with cortical bone, while new bone formation was only observed in the ends of artificial bone and there were not continuous bone callus on the surface in the control side. CONCLUSION: Artificial bone with PRP is effective in the repair of segmental bone defects, and PRP could improve the healing of bone defect.
Objective To explore the effect of the platelet-rich plasma (PRP) on proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells (MSCs) in China goat in vitro. Methods MSCs from the bone marrow of China goat were cultured. The third passage of MSCs were treated with PRP in the PRP group (the experimental group), but the cells were cultured with only the fetal calf serum (FCS) in the FCS group (the control group). The morphology and proliferation of the cells were observed by an inverted phase contrast microscope. The effect of PRP on proliferation of MSCs was examined by the MTT assay at 2,4,6 and 8 days. Furthermore, MSCs were cultured withdexamethasone（DEX）or PRP; alkaline phosphatase (ALP) and the calcium stainingwere used to evaluate the effect of DEX or PRP on osteogenic differatiation of MSCs at 18 days. The results from the PRP group were compared with those from the FCS group. Results The time for the MSCs confluence in the PRP group was earlier than that in the FCS group when observed under the inverted phase contrast microscope. The MTT assay showed that at 2, 4, 6 and 8 days the mean absorbance values were 0.252±0.026, 0.747±0.042, 1.173±0.067, and 1.242±0.056 in the PRP group, but 0.137±0.019, 0.436±0.052, 0.939±0.036, and 1.105±0.070 in the FCS group. The mean absorbance value was significantly higher in the PRP group than in the FCS group at each observation time (P＜0.01). Compared with the FCS group, the positive-ALP cells and the calcium deposition were decreased in the PRP group; however, DEX could increase boththe number of the positiveALP cells and the calcium deposition. Conclusion The PRP can promote proliferation of the MSCs of China goats in vitro but inhibit osteogenic differentiation.
Objective To study the mechanism of compound of calcium phosphate(TCP) and platelet-rich plasma(PRP) in the treatment of femoral head necrosis.Methods The left femoral heads of 48 New Zealand white rabbits were frozen by liquid nitrogen as to make themodel of femoral head necrosis.Twenty-four rabbits were randomly chosen as theexperimental group and their femoral heads were filled with TCP/PRP. The other 24 rabbits were used as the control group and their femoral heads were filled only with TCP. They were sacrificed at 2, 4,8,12 weeks after operation. The specimens were examined with X-ray and histological study.Results At 2 weeks after operation,there was no significant difference in femoral headdensity between the two groups. Four weeks after operation, femoral head density decreased in both groups, while it decreased more in the control group. At 8,12 weeks after operation, the density of the femoral heads in both groups increased, and it was higher in the experimental group. Histology examination showed thatthere was no difference between the two groups 2 weeks after operation. The head became flat at 4 weeks. Control group had more defects. At 4,8,12 weeks, more repairs were observed in the experimental group than that in the control group. The amount and maturity of osteogenesis in experimental group were much more greaterthan those in control group．Bone histomorphometry showed that the volum of thetrabecular was larger in the experimental group (36.65%±7.22%,38.29%±4.28%，39.24%±3.42%) than that of control group(P＜0.05). Conclusion TCP/PRP does not only provide osteoblasts scaffold, butalso promotes bone formation and the head repair. TCP/PRP is a good biomaterialfor the treatment of femur head necrosis.
ObjectiveTo explore the relationship of platelet-activating factors and vascular endothelial activity markers to lacunar infarction (LI).MethodsA total of 100 inpatients diagnosed with LI in Shaanxi Provincial People’s Hospital between March 2018 and February 2019 were included, and 100 matched healthy controls were collected. Basic information, clinical baseline data, laboratory examinations, cerebral MRI and treatment data were collected after admission. The platelet-activating factors (platelet membrane glycoprotein Ⅱb/Ⅲa receptor and P-selectin) and vascular endothelial activity markers [von Willebrand factor (vWF), homocysteine (HCY), and high-sensitivity C-reactive protein (hsCRP)] levels of patients with LI were detected one month and three months after onset, and those of the control group were decteted when they were selected. SPSS 25.0 software was used for statistical analysis.ResultsAt one month after onset, there was no statistically significant difference in the levels of platelet activating factors between the LI group and the control group [platelet membrane glycoprotein Ⅱb/Ⅲa receptor: (2.84±1.00)% vs. (2.59±0.96)%, P=0.065; P-selectin: (3.05±0.63)% vs. (2.98±0.59)%, P=0.419], while the differences in the levels of vascular endothelial activity markers between the two groups were statistically significant [vWF: (141.80±17.60) vs. (124.63±10.65) ng/mL, P<0.001; hsCRP: (5.53±1.37) vs. (2.17±0.55) mg/L, P<0.001; HCY: (18.76±4.07) vs. (15.81±2.63) mmol/L, P<0.001]. At three months after onset, 94 LI patients were followed up. The levels of vWF and hsCRP between the 100 patients one month after onset and the 94 patients three months after onset were statistically different [(vWF: (141.80±17.60) vs. (134.86±13.35) ng/mL, P=0.002; hsCRP: (5.53±1.37) vs. (2.63±0.55) mg/L, P<0.001], but there was no statistically significant difference between the two time points in the levels of HCY or platelet-activating factors (P>0.05).ConclusionChronic platelet activation may not play a core role in LI pathophysiology, and endothelial dysfunction may be one of the pathological mechanisms of LI.
目的 探讨脂多糖（LPS）、白细胞介素-6（IL-6）和血小板活化因子（PAF）与重症胸腹创伤后凝血功能紊乱发生的相关性及可能的致病机理。方法 收集2009年1月至2011年12月期间在中国人民解放军第二五三医院急诊科就诊、创伤指数≥17分且除外合并颅脑损伤及在急诊科内死亡的胸腹创伤患者62例，在予以抢救、治疗的同时抽血检查血小板计数（PLT）、血浆D-二聚体（D-D）、部分活化凝血酶原时间（APTT）、凝血酶原时间（PT）、LPS、IL-6和PAF，并对其结果进行相关性分析。结果 本组患者就诊时检测的PLT为（157.73±78.11）×109/L， D-D为（1 023.88±208.72） U/L，APTT为（46.95±17.85） s，PT为（19.44±6.95） s，TT为（58.27±12.44）s，除PLT降低外，其余4项指标均升高或延长； LPS为（322.85±104.54） U/L，IL-6为（285.51±81.46） ng/mL，PAF为 （14 714.70±4 427.95） ng/L， 三者均升高； PLT与LPS、IL-6和PAF之间呈负相关关系（P＜0.001），而D-D、APTT、PT和TT与LPS、IL-6和PAF之间均呈正相关关系（P＜0.001）。结论 LPS、IL-6及PAF可能参与了重症胸腹创伤后凝血功能障碍的发生；重症胸腹损伤后出现的微循环障碍及内毒素血症是凝血功能障碍发生的重要机理。针对LPS、IL-6和PAF进行早期干预，有可能改善重症胸腹创伤患者的凝血功能障碍。
ObjectiveTo explore the clinical manifestations, pathogenesis, prognosis and the therapeutic modalities of the patients with solid tumors combined with remarkable hematologic abnormalities. MethodsGastric cancer in a paitnet was diagnosed by endoscopy as well as pathological biopsy and myeloproliferative neoplasms (MPNs) were excluded through bone marrow examinations. Therefore, the primary and metastatic malignancies were excised. ResultsAlthough surgical operation improved the hematologic changes, the high tumor load caused the poor prognosis. ConclusionThe solid tumors may present with hematologic manifestations which is similar to the symptoms of MPNs; it belongs to the para-neoplastic phenomenon and may be an independent poor prognostic factors.
Objective To study the effects of platelet-rich plasma(PRP) on the proliferation and osteogenetic activity of the marrow mesenchymal stem cells(MSCs) cultured in vitro to elucidate the cellular and molecularmechanism by which PRP accelerates bone repair.Methods The human MSCs were cultured in vitro and randomly divided into the experimental group(n=9) and control group(n=9). In the experimental group, the MSCs were interfused with PRP(10 μl/ml culture media). The proliferation ability of the cells was tested by flow cytometry and MTT, the osteogenetic activity by alkaline phosphatase(ALP) measuring and tetracycline fluorometry, and cbfal mRNA expression by reverse transcriptPCR.Results PRP could stimulate the MSCs proliferation. The flow cytometry assay showed that the MSCs proportion of S period of the experimental group significantly increased 14.5±0.4 in comparison with that of the control group 7.2±0.5 (P＜0.01) after 24 hours. MTT value showed that MSCs proliferatedto platform period earlier in the experimental group than in the control group. There was a significant increase in ALP activity of the experimental group 7.79±1.98,9.51±2.31and 14.03±3.02 when compared with that of the control group 2.06±0.77,2.84±0.82 and 2.58±0.84 after 3, 6 and 9 days(P＜0.05). The number of mineral nodes increased. Reverse transcript-PCR showed that the expression of cbfal mRNA were elevated gradually at 2,4 and 8 hours after interfused with PRP.Conclusion The effect of PRP on accelerating bone repair is related to its effects on stimulating the proliferation of MSCs, increasing the cbfal expression and promoting the osteogenetic activity.