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 investigate the interleukin-17 (IL-17) levels changes in both synovial fluid and venous plasma of patients with primary knee osteoarthritis (OA) after intra-articular injection of platelet-rich plasma (PRP). Methods Between January 2015 and January 2016, 30 patients with primary knee OA were treated by intra-articular injection of PRP once a week for 3 weeks (trial group). Thirty healthy individuals were recruited into the study as control. There was no significant difference in gender, age, and body mass index between 2 groups (P>0.05). Visual analogue scale (VAS) score and Knee Society Score (KSS) were used to evaluate pain level and function of the knee for patients with OA. The IL-17 levels in both venous plasma and synovial fluid were measured before injection and at 1, 3, 6, and 12 months after injection in trial group and the IL-17 levels in venous plasma were measured in control group. The levels were determined using ELISA method. Results There was no knee joint swelling, fever, local infection, or other uncomfortable symptoms for all patients in process of PRP injection. All patients were followed up 13.5 months on average (range, 12-15 months). In trial group, the VAS scores at different time points after injection were significantly lower than that before injection (P<0.05). And the KSS scores at different time points after injection were significantly higher than that before injection (P<0.05). There was no significant difference in VAS and KSS scores between different time points after injection (P>0.05). The IL-17 levels in venous plasma before and after injection in trial group were significantly higher than that in control group (P<0.05). The IL-17 levels in venous plasma at each time point after injection were significantly lower than that before injection (P<0.05). There was no significant difference in IL-17 levels in both venous plasma and synovial fluid between different time points after injection (P>0.05). Conclusion Intra-articular injection of PRP can significantly release the pain symptoms, improve joint function, and reduce IL-17 levels in both synovial fluid and venous plasma of the patients with knee OA, but IL-17 levels can not reduce to normal level.
Perioperative monitoring of blood coagulation is critical to better understand causes of hemorrhage, to guide hemostatic therapies, and to predict the risk of bleeding. Point-of-care (POC) coagulation monitoring devices assessing the viscoelastic properties of whole blood may overcome several limitations of routine coagulation tests in the perioperative setting. The advantage of these techniques is that they have the potential to measure the clotting process, starting with fibrin formation and continue through to clot retraction and fibrinolysis at the bedside, with minimal delays. Furthermore, the coagulation status of patients is assessed in whole blood, allowing the plasmatic coagulation system to interact with platelets and red blood cells, and thereby providing useful additional information on platelet function. Viscoelastic POC coagulation devices are increasingly being used in clinical practice, especially in the management of patients undergoing cardiac and liver surgery, assessment of hypo-and hypercoagulable states, guiding pro- and anticoagulant therapies, monitoring of antiplatelet therapy and procoagulant therapy. To ensure optimal accuracy and performance, standardized procedures for blood sampling and handling, strict quality controls and trained personnel are required.
ObjectiveTo identify and observe the pathogenic genes and clinical phenotypes of a family with a special platelet phenotype, Hermansky-Pudlak syndrome type 6 (HSP6). MethodsA retrospective clinical study. In November 2019, one proband and three family members from six HSP families who visited Henan Eye Hospital were included in the study. The child's medical history and family history were inquired in detail. The proband and all family members underwent best corrected visual acuity (BCVA), fundus color photography, frequency-domain optical coherence tomography, and general physical examination. The proband underwent platelet transmission electron microscopy (PTEM) and colonoscopy. Peripheral venous blood was collected from the proband, her parents and younger brother, and genomic DNA was extracted. Whole exome sequencing (WES) was used to screen pathogenic genes and their loci. Bioinformatics analysis determines the pathogenicity of gene variation sites. Fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to verify the related variations. ResultsThe proband (Ⅱ-1) was a 7-year-old female. The BCVA in both eyes was 0.1, who exhibited mild horizontal nystagmus and iris depigmentation. Fundus examination revealed obvious depigmentation and an underdeveloped fovea centralis. At the age of 7, the patient underwent colonoscopy due to acute gastrointestinal bleeding. A polyp approximately 5 mm in size was found on the floor of the sigmoid colon, with erosion and mucosal leukoplakia on its surface. PTEM showed that the number of platelet dense granules was normal, but the nuclei were small or exhibited low compactness. The skin on both lower legs showed pigmentation. The clinical phenotypes of the proband’s parents (Ⅰ-1, Ⅰ-2) and younger brother (Ⅱ-2) showed no obvious abnormalities. WES revealed that the proband carried compound heterozygous variants in exon 1 of the HPS6 gene: c.60_64dup (p.L22fs) (M1) and c.1147_1148del (p.L383fs) (M2). The mother carried the M1 variant, while the father and younger brother carried the M2 variant. Bioinformatics analysis predicted that both variants were pathogenic. RT-qPCR results showed that, compared with the relative expression level of HPS6wt mRNA, the relative expression levels of HPS6L22fs and HPS6L383fs mRNA were significantly decreased (t = 3.549, 4.560; P＜0.05). Western blot analysis demonstrated that the HPS6L383fs protein was truncated, whereas the HPS6L22fs protein was not detected. ConclusionsThis family is a special HPS6 with a normal number of dense platelet granules. The compound heterozygous variations of M1 and M2 in the HPS6 gene are pathogenic genes in this family.
Objective To compare the difference of the changes of platelet counts after splenectomy between the patients with splenic rupture and patients with cirrhosis and portal hypertension, and to analyze the possible reasons and clinical significance. Methods The platelet count of 47 splenic rupture patients and 36 cirrhosis patients who had been carried out splenectomy from July 2008 to December 2009 in our hospital were counted, and the differences in platelet count and it’s change tendency of two groups were compared. Results In the splenic rupture group，the platelet count of all 47 patients increased abnormally after operation, the maxlmum value of platelet count among 300×109/L-600×109/L in 6 cases，600×109/L-900×109/L in 21 cases，and above 900×109/L in 20 cases. In the cirrhosis group，the maxlmum value of platelet count after operation was above 300×109/L in 26 cases，100×109/L-300×109/L in 8 cases，and below 100×109/L in 2 cases. The difference of maxlmum value of platelet count in the two groups had statistic significance（P=0.00）. Compared with the cirrhosis group, the platelet count increased more significant and decreased more slow in splenic rupture group（P＜0.05）.The abnormal days and rising range of platelet count were higher in patient with Child A than Child B and C（P=0.006，P=0.002）. Conclusions The change of platelet count after operation in splenic rupture group was obviously different from cirrhosis group because of the difference of the liver function and body situation of patients. To patients with splenic rupture or cirrhosis, appropriate treatment based on the platelet count and liver function could obtain good therapeutic effect.
Objective Platelet-rich plasma (PRP) can enhance the chondrocyte prol iferation and repair of cartilage defects. To explore the safety and efficacy of intra-knee-articular injection of PRP to treat knee articular cartilage degeneration by comparing with injecting sodium hyaluronate (SH). Methods Thirty consecutive patients (30 knees) with knee articular cartilage degeneration were selected between January 2010 and June 2010. According to different injections, 30 patients wererandomly divided into PRP group (test group, n=15) and SH group (control group, n=15). There was no significant difference in gender, age, body mass index, and Kellgren-Lawrence grade between 2 groups (P gt; 0.05). Test group received 3.5 mL of PRP intra-knee-articular injections while control group received 2 mL of SH during the same time period. Both treatments were administered in series of 3 intra-knee-articular injections at 3-week intervals. Then, adverse reactions were recorded. International Knee Documentation Committee (IKDC) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and Lequesne index were used for evaluation of treatment results. Results The patients of 2 groups were followed up 6 months. There were significant differences in IKDC score, WOMAC score, and Lequesne index between pre- and post-injection in 2 groups (P lt; 0.05); no significant difference was found between different time points (3, 4, and 6 months) in test group (P gt; 0.05), while significant differences were found between the postoperative 6th month and the postoperative 3rd and 4th months in control group (P lt; 0.05). There was no significant difference in IKDC score, WOMAC score, and Lequesne index between 2 groups within 4 months (P gt; 0.05), but the effectiveness of test group was significantly better than that of control group at 6 months after injection (P lt; 0.05). Adverse reactions occurred in 12 patients (31 injections) of test group and in 12 patients (30 injections) of control group. No significant difference in onset time, termination time, and duration of adverse reactions were found between 2 groups (P gt; 0.05). Conclusion Intra-knee-articular injection of PRP to treat knee articular cartilage degeneration is safe, which can alleviate symptoms of pain and swell ing and improve the qual ity of l ife of patients; however, further data of large samples and long-term follow-up are needed to confirm the safety and effectiveness.
Objective To investigate the expressions of platelet derived growth factor (PDGF) and survivin in hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) and to evaluate the relationships among the expressions of PDGF, survivin and the proliferation of cancer cells. Methods The expression of PDGF mRNA in 16 cases in HCC with PVTT group was observed by in situ hybridization and the results were compared with that in HCC tissue group. The expressions of PDGF and survivin protein in 36 cases in HCC with PVTT group were detected with immunohistochemistry and it was found that there was a correlation between them. Flow cytometry (FCM) was used to measure the proliferation of cancer cells and it was also used to analyze the relations among PDGF, survivin and the proliferation of cancer cells. Results The expression level of PDGF mRNA in HCC with PVTT group was significantly higher than that in HCC tissue group (P＜0.01). There was a positive correlation between the expressions of PDGF and survivin protein in HCC with PVTT (P＜0.01). The degree of proliferation of cancer cells in PDGF and survivin protein positive group was significantly higher than that in negative group (P＜0.01). Conclusion PDGF and survivin gene over-expressed in HCC with PVTT group and there is a positive correlation between the expressions of PDGF and survivin protein. The proliferation of cancer cells increases as the expressions of PDGF and survivin increase.
Objective The use of autologous platelet-rich gel (APG) is a relatively new technology and a promising treatment method for infections, which is currently being used by a variety of surgical specialties. The mechanism of antibacterialeffect of APG is not yet fully discovered. Subsequent evidence suggests that platelets have multi ple functional attributes inantimicrobial host defense (including the capacity to generate antimicrobial oxygen metabol ites and the antimicrobial peptides) and interact directly with microorganisms, contribute to clearance of pathogens from the blood. To investigate the bacteriostasis of APG against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa in vitro. Methods Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were obtained from whole blood of 17 healthy donors. APG was prepared by mixing PRP with bovine thrombin in a 10% calcium gluconate solution or bovine thrombin in a 10% calcium gluconate solution and apocynin (APG-APO). Antibacterial effects of APG, PRP, and APG-APO on Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were evaluated by bacteriostasis assay. Results The culture results showed apparent decrease in the number of Staphylococcus aureus for both APG and APG-APO, which was maximal at first 4 hours and lasted to 24 hours and 8 hours, respectively; showing significant difference (P lt; 0.05) when compared APG with PRP and PPP, however no significant difference at first 8 hours (P gt; 0.05) and significant difference at 12 and 24 hours (P lt; 0.05) when compared APG with APG-APO; showing significant difference at first 4 hours (P lt; 0.05), no significant difference at 6, 8, 12, and 24 hours when compared APG-APO with PRP and PPP (P gt; 0.05). The bacteriostasis rates of APG and APG-APO were 27.36%-52.97% and 18.82%-51.52% against Escherichia coli, respectively; showing no significant difference (P gt; 0.05) when compared with PRP. The bacteriostasis rates of APG and APG-APO were less than 35% against Pseudomonas aeruginosa, showing no significant difference (P gt; 0.05) when compared with PRP; the bacteriostasis rates of PRP were less than 15% against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Conclusion APG may have potential bacteriostatic effect against Staphylococcus aureus by platelet mediating. Either APG or APG-APO has no obvious bacteriostatic effect against Escherichia coli or Pseudomonas aeruginosa. PRP has no antibacterial activity against Staphylococcus aureus, Escherichia coli or Pseudomonas aeruginosa.
ObjectiveTo investigate the predictive factors of portal vein thrombosis (PVT) before and after splenectomy and gastroesophageal devascularization for liver cirrhosis with portal hypertension. MethodsSixty-one cases of liver cirrhosis with portal hypertension who underwent splenectomy and gastroesophageal devascularization were enrolled retrospectively. The patients were divided into PVT group and non-PVT group based on the presence or absence of postoperative PVT on day 7. The clinical factors related with PVT were analyzed. ResultsThere were 25 cases in the DVT group and 36 cases in the non-DVT group. The results of univariate analysis showed that the preoperative platelet (P=0.006), activated partial thromboplastin time (P=0.048), prothrombin time (P=0.028), and international normalized ratio (P=0.029), postoperative fibrin degradation product (P=0.002) and D-dimer (P=0.014) on day 1, portal venous diameter (P=0.050) had significant differences between the DVT group and non-DVT group. The results of logistic multivariate regression analysis showed that the preoperative platelet (OR=0.966, 95% CI 0.934-1.000, P=0.048) and postoperative fibrin degradation product on day 1(OR=1.055, 95% CI 1.011-1.103, P=0.017) were correlated with the PVT. The PVT might happen when preoperative platelet was less than 34.5×109/L (sensitibity 80.6%, specificity 60.0%) or postoperative fibrin degradation product on day 1 was more than 64.75 mg/L (sensitibity 48.0%, specificity 91.7%). ConclusionPreoperative platelet and postoperative fibrin degradation product on day 1 might predict PVT after splenectomy and gastroesophageal devascularization for liver cirrhosis with portal hypertension.
To introduce the current situation of platelet-rich plasma (PRP) and its appl ication in orthopedics field. Methods The latest l iterature was reviewed, concerning the preparations of PRP, physiological mechanism and the latest appl ications in orthopedics field. Results PRP represent a new biotechnology for the stimulation and acceleration of tissue heal ing and bone regeneration. To succeed, some of the challenges need to be addressed including standardization and definition of the different PRP preparations, the complete characterization of the platelet released factors and proteins. Furthermore, well-designed studies and cl inical trials are needed to evaluate the potential therapeutic impact ofPRP. Conclusion PRP provides a novel appl ication for orthopedics field.