Effect of autologous serum after amniotic membrane transplantation for persistent corneal ulcers

AIM:To investigate the effect of adding autologous serum eye drops to the postoperative regime after amniotic membrane transplantation for severe persistent corneal ulcers.METHODS:Forty eyes of 40 patients with persistent corneal ulcers were randomly assigned to artificial tears(sodium hyaluronate 0.2%,ATs group,n=20)or autologous serum eye drops(ASEDs,n=20)following treatment with amniotic membrane transplantation.Digital slit lamp images were acquired from all patients before and 30d post treatment.The area with fibrovascular tissue was calculated using Image J.Central corneal sensitivity was assessed by Cochet-Bonnet aesthesiometry before and one month after treatment.Scar tissue transparency was assessed with a novel optical densitometry.RESULTS:Mean age of patients was 61.65±16.47y and 57.3±19.11y in the ATs group and ASEDs group,respectively.Twenty-two male and 18 female patients were included in the study.The improvement in visual acuity was significantly greater in the ASEDs group(0.14±0.04)than the ATs(0.08±0.04;P=0.00046).Cochet-Bonnet aesthesiometry improved significantly after treatment with a similar rate between groups.There were no statistically significant differences in the area of postoperative fibrovascular tissue between the two groups(P=0.082).The success rate in the two groups was similar.The difference in densitometry between the ATs and ASEDs group was statistically significant(P=0.042)with greater reduction from baseline in the ASEDS group.CONCLUSION:Autologous serum eye drops can lead to better visual acuity,more stable results and improved densitometry and should be considered in the postoperative care following amniotic membrane transplantation.

Conservative management of spinal pathology with autologous conditioned serum: A systematic review of the literature

BACKGROUND Chronic inflammatory pain is associated with increased expression of interleukin(IL)-1,an inflammatory cytokine,and activity on its receptor(IL-1R).In response,the body produces IL-1R antagonist(IL-1Ra)to reduce this signaling.Autologous conditioned serum(ACS)is the only biologic therapy for spinal pathologies that enhances the action of endogenous IL-1Ra reserves to improve symptoms.This systematic review investigates the effectiveness of ACS in treating pain and dis-ability caused by spinal pathologies.AIM To evaluate the use of ACS as a conservative management option for spinal path-ology.METHODS A systematic review of PubMed/Medline was performed to identify studies inve-stigating administration of ACS for treatment of any spinal pathology.RESULTS Six articles were included,comprising 684 patients treated with epidural(n=133)or transforaminal(n=551)ACS injections.Patients had an average age of 54.0 years with slight female predominance(53.2%).The lumbar spine was most com-monly treated,with 567 patients(82.9%)receiving injections for lumbar radicu-lopathy(n=67),degenerative disc disease(DDD)(n=372),or spinal stenosis(n=128);cervical injections were performed in 109 patients(15.9%).Mean(SD)follow-up was 21.7(4.8)weeks from first ACS injection.All studies investigating mecha-nical lumbar and lumbar or cervical radicular pain reported significant pain re-duction at final follow-up compared to baseline.ACS achieved comparable or su-perior results to lumbar epidural steroid injections.Adverse events were reported in 21 patients(3.1%),with no serious adverse events.CONCLUSION ACS injection is a safe and effective intervention for pain reduction in many spinal pathologies,including cervical and lumbar radiculopathies.

Effect of human autologous serum and fetal bovine serum on human corneal epithelial cell viability,migration and proliferation in vitro

AIM： To analyze the concentration-dependent effects of autologous serum （AS） and fetal bovine serum （FBS） on human corneal epithelial cell （HCEC） viability, migration and proliferation. METHODS： AS was prepared from 13 patients with non- healing epithelial defects Dulbecco＇s modified eagle medium/ Ham＇s F12 （DMEM/F12） with 5% FBS, 0.5% dimethyl sulphoxide （DMSO）, 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media： DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS. HCEC viability was analyzed using cell proliferation kit XTI＇, migration using a wound healing assay, proliferation by the cell proliferation enzyme-linked immunosorbent assay （ELISA） BrdU kit. Statistical analysis was performed using the generalized linear model, the values at 30% AS or 30% FBS were used as the baselines. RESULTS： HCEC viability was the highest at 30% AS or 15% FBS and the lowest at 10% AS or 30% FBS application. HCEC migration was the quickest through 30% AS or 30% FBS and the slowest through 5% AS or 5% FBS concentrations. Proliferation was the most increased through 15% AS or 5% FBS and the least increased through 30% AS or 30% FBS concentrations. HCEC viability at 10% and 15% AS was significantly worse （P=0.001, P=0.023） compared to baseline and significantly better at 15% FBS （P=0.003） concentrations. HCEC migration was significantly worse （P〈0.007） and HCEC proliferation significantly better （P〈0.001） in all concentration groups compared to baseline. CONCLUSION： For the best viability of HCEC 30% AS or 15% FBS, for HCEC migration 30% AS or 30% FBS, for proliferation 15% AS or 5% FBS should be used. Therefore, we suggest the use of 30% AS in clinical practice.

Platelet-rich plasma increases transforming growth factor-beta1 expression at graft-host interface following autologous osteochondral transplantation in a rabbit model

AIM: To explore the effect of platelet-rich plasma on protein expression patterns of transforming growth factor-beta1(TGF-β1) in cartilage following autologous osteochondral transplantation(AOT) in a rabbit knee cartilage defect model.METHODS: Twelve New Zealand white rabbits received bilateral AOT. In each rabbit, one knee was randomized to receive an autologous platelet rich plasma(PRP) injection and the contralateral knee received saline injection. Rabbits were euthanized at 3, 6 and 12 wk post-operatively. Articular cartilage sections were stained with TGF-β1 antibody. Histological regions of interest(ROI)(left, right and center of the autologous grafts interfaces) were evaluated using Meta Morph. Percentage of chondrocytes positive for TGF-β1 was then assessed.RESULTS: Percentage of chondrocytes positive for TGF-β1 was higher in PRP treated knees for selected ROIs(left; P = 0.03, center; P = 0.05) compared to control and was also higher in the PRP group at each post-operative time point(P = 6.6 × 10^(-4), 3.1 × 10^(-4) and 7.3 × 10^(-3) for 3, 6 and 12 wk, respectively). TGF-β1 expression was higher in chondrocytes of PRP-treated knees(36% ± 29% vs 15% ± 18%)(P = 1.8 × 10^(-6)) overall for each post-operative time point and ROI. CONCLUSION: Articular cartilage of rabbits treated with AOT and PRP exhibit increased TGF-β1 expression compared to those treated with AOT and saline. Our findings suggest that adjunctive PRP may increase TGF-β1 expression, which may play a role in the chondrogenic effect of PRP in vivo.

Keratinocyte growth factor-2 and autologous serum potentiate the regenerative effect of mesenchymal stem cells in cornea damage in rats

AIM:To investigate the healing process after severe corneal epithelial damage in rats treated with mesenchymal stem cells(MSCs)cultured with or without keratinocyte growth factor(KGF-2)and autologous serum(AS)on amniotic membrane(AM).Many patients are blind and devastated by severe ocular surface diseases due to limbal stem cell deficiency.Bone marrow-derived MSCs are potential sources for cellbased tissue engineering to repair or replace the corneal tissue,having the potential to differentiate to epithelial cells.METHODS:The study included 5 groups each including 10 female'Sprague Dawley'rats in addition to20 male rats used as bone marrow donors.Group I rats received AM+MSCs,Group II rats AM+MSCs cultured with KGF-2,Group III rats AM+MSCs cultured with KGF-2+AS,Group IV rats only AM and Group V rats,none.AS was derived from blood drawn from male rats and bone marrow was obtained from the femur and tibia bones of the same animals.Therapeutic effect was evaluated with clinical,histopathological and immunohistochemical assessment.MSC engraftment was demonstrated via detection of donor genotype(Y+)in the recipient tissue(X)with polymerase chain reaction.RESULTS:Corneal healing was significantly better in Groups I-III rats treated with MSC transplantation compared to Group IV and Group V rats with supportive treatment only.The best results were obtained in Group III rats with 90%transparency,70%lack of neovascularization,and 100%epithelium damage limited to less than 1/4 of cornea.CONCLUSION:We suggest that culture of MSCs with KGF-2 and AS on AM is effective in corneal repair in case of irreversible damage to limbal stem cells.

Autologous peripheral blood-derived orthobiologics:Different types and their effectiveness in managing knee osteoarthritis

Knees are the most commonly impacted weight-bearing joints in osteoarthritis(OA),affecting millions of people worldwide.With increasing life spans and obesity rates,the incidence of knee OA will further increase,leading to a significant increase in the economic burden.Conventional treatment modalities utilized to manage knee OA have limitations.Over the last decade,the role of various autologous peripheral blood-derived orthobiologics(APBOs)for the treatment of knee OA has been extensively investigated.This editorial provided an overview and focused on defining and shedding light on the current state of evidence based on the most recent published clinical studies concerning the use of APBO for the management of knee OA.While numerous studies have demonstrated promising results for these preparations,a notable gap exists in the comparative analysis of these diverse formulations.This absence of head-to-head studies poses a considerable challenge for physicians/surgeons in determining the optimal preparation for managing knee OA and achieving sustained longterm results.Thus,more adequately powered,multicenter,prospective,doubleblind,randomized controlled trials with longer follow-ups are needed to establish the long-term efficacy and to aid physicians/surgeons in determining the optimal APBO for the management of knee OA.

Randomized controlled trial on the efficacy and safety of autologous serum eye drops in dry eye syndrome

BACKGROUND Autologous serum eye drops(ASEDs),a novel treatment derived from blood serum,have emerged as a groundbreaking solution for managing dry eye syndrome(DES).These drops have shown significant promise in relieving the distressing symptoms of DES.This study aimed to evaluate the safety and effectiveness of ASEDs compared to traditional treatments,which often prove inadequate or result in unwanted side effects,particularly in individuals with moderate-to-severe DES.AIM To evaluate whether ASEDs are safer and more effective than conventional artificial tears in the treatment of moderate-to-severe DES.METHODS This multi-centered randomized controlled trial included 240 patients with moderate-to-severe DES from three ophthalmology clinics in China.They were randomly assigned to receive either ASEDs or artificial tears for 12 wk.The primary outcome was the change in the ocular surface disease index(OSDI)score,with secondary outcomes including tear break-up time(TBUT),Schirmer I test,corneal fluorescein staining(CFS),and conjunctival impression cytology(CIC).Statistics analysis was performed using an analysis of covariance with adjustments made for baseline values.RESULTS Our findings revealed that both ASEDs and artificial tears significantly improved the OSDI score,TBUT,Schirmer I test,CFS,and CIC from baseline to week 12.The ASEDs group showed significantly greater improvement in all these measures than the artificial tears group(all P values<0.05).The average difference in the OSDI score between the two cohorts was-10.3(95%confidence interval:-13.6 to-7.0),indicating a substantial improvement in the ASEDs group.The occurrence of adverse events was comparable between cohorts,with no reports of severe adverse events.CONCLUSION ASEDs are more effective and safer than artificial tears for mitigating symptoms of moderate-to-severe DES.ASEDs could be an alternative/supplementary therapy for patients with DES less responsive to traditional treatments.

Autologous serum can induce mesenchymal stem cells into hepatocyte-like cells

Objective： To investigate whether the rabbit serum after radiofrequency ablation to liver tumor can induce mesenchymal stem cells （MSCs） differentiating into hepatocyte-like cells in order to find a new source and culture process for repairing liver injury. Methods： A tumor piece of 1 mm× 1mm×1 mm was transplanted into a tunnel at right liver of rabbits. The model of liver tumor was established after 2-3 weeks. The serum was collected from rabbits 72 h after being subjected to radiofrequency ablation of the liver tumor. Mesenchymal stem cells were isolated from rabbit bone marrow and cultured in DMEM containing autologous rabbit serum. Three kinds of media （L-DMEM） were tested respectively： ① containing 10% fetal calf serum （FCS）; ② containing 30% rabbit autologous serum after radiofrequency ablation of the liver tumor （ASRF）; ③ containing 30% rabbit autologous serum （AS）. MSCs were cultured on 12-well plates until passage 2 and examined under the light and electron microscopy at indicted intervals. The expression of albumin and CKl8 was detected using immunofluorescence to identify the characteristics of differentiated cells. Results： MSCs performed differently in the presence of fetal calf serum, rabbit autologous serum and rabbit autologous serum after radiofrequency ablation of the liver tumor. Induced by the serum after radiofrequency ablation to liver tumor for 7 d, the spindle-shaped MSCs turned into round shaped and resembled hepatocyte-like cells. The reactions were not found in MSCs cultured in FCS and AS groups. After induction for 14 d, slender microvilli, cell-cell junction structure and cholangiole emerged, and the differentiated cells expressed albumin and CKl 8. All those could not been observed in 10% FCS and 30% autologous serum groups. Conclusion： Mesenchymal stem cells differentiate into hepatocyte-like cells in the serum after radiofrequency ablation of liver tumor, providing us a potential cell source and culture process for clinical application in liver injury repairing.

Differentiation of Mesenchymal Stem Cells into Hepatocyte-Like Cells by Autologous Serum after Radiofrequency Ablation of Liver Tumor

Mesenchymal stem cells?(MSCs) have been shown to differentiate into liver cells in serum of part-resection liver, but it was hardly feasible in clinical use. Our studies revealed that MSCs could differentiate into hepatocyte-like cells in autologous serum after radiofrequency ablation (RFA) therapy of the liver tumor. Rabbits with liver tumor subsequently treated with RFA therapy. Serum was collected from those rabbits before RFA therapy and 72 hours after RFA therapy. MSCs were isolated from each rabbit’s bone marrow and cultured in DMEM medium containing the following different supplements: 30% fetal calf serum (FCS group), 30% rabbit autologous serum (AS group) or 30% autologous serum after RFA treatment of the liver tumor (ASRF group), observed by electron microscopy, flow cytometry, immunofluorescence. Seven days later, most of the spindle-shaped MSCs in the ASRF group transformed into polygon or round-shaped cells resembling hepatocytes, and the percentage in S/G2/M phase was higher than in the FCS or AS groups. Fourteen days later, slender microvilli, cell-cell junction structures and cholangiole emerged in the cells belonging to the ASRF group, the expression of albumin and CK18 was observed only in the differentiated cells from the ASRF group. These changes were not observed in the FCS group or the AS group. This study may provide a potential cell source and culture process for clinical application in liver injury treatment.

Clinical and Magnetic Resonance Image Changes of Platelet-Rich Plasma Therapy in Combination with Human Mesenchymal Stem Cells from Autologous Adipose Tissue for Knee Osteoarthritis Treatment

Objective: To evaluate the efficacy based on clinical symptom and on magnetic resonance image of platelet-rich plasma therapy in combination with mesenchymal stem cells from autologous adipose tissue for knee osteoarthritis treatment. Patients and Method: 30 patients including 26 females and 4 males;correspondingly, 60 knee joints were diagnosed with osteoarthritis with stages II - III of Kellgren and Lawrence, their mean age was 58.63 ± 11.11. All were injected with autologous platelet-rich plasma that was extracted by PRP set, APC 30 PRP PRCEDURE PRAK and autologously extracted mesenchymal stem cells from abdominal adipose tissue using the ADI-25-01 ADIPOSEPRCEDURE PRAK of USA. Results: After 12 months: the pain level according to VAS score at the right knee joint was decreased from 6.0 ± 1.28 before treatment to 1.9 ± 0.3;VAS score at the left knee joint was decreased from 6.43 ± 1.19 to 2.25 ± 0.43. Total Lequene score at right knee joint was decreased from 16.04 ± 1.57 before treatment to 4.31 ± 1.04, at left knee joint was decreased from 17.52 ± 1.74 before treatment to 5.15 ± 1.48. Total WOMAC score at right knee joint was decreased from 55.93 ± 5.56 to 10.37 ± 1.56;at left knee joint was decreased from 53.97 ± 5.57 to 10.07 ± 1.59. There were 86.77% joints with cartilage thickness change and the patellar cartilage thickness was increased from 1.56 ± 0.09 mm before treatment to 1.65 ± 0.09 mm. Conclusion: The treatment of knee osteoarthritis by platelet-rich plasma therapyin combination with mesenchymal stem cells from autologous adipose tissue is effective in reducing pain, improving patient's mobility and walking function, reforming articular cartilage thickness on magnetic resonance image.

Immunologic changes to autologous transfusion after operational trauma in malignant tumor patients:Neopterin and Interleukin-2

Objective:To estimate the impact of autologous transfusion on the status of perioperative immune activation inmalignant tumor patients.The Serum Neopterin and Interleukin-2(IL-2)were measured.Methods:Sixty patients undergoingelective radical resection for malignant stomach tumor were enrolled in the prospective study and assigned to the following groups:(Ⅰ)Group A received autologous transfusion;(2)Group H received allogeneic transfusion.The perioperative course(Beforeinduction of anesthesia,after operation and 5 d after operation)of Neopterin and IL-2 was compared.Results:In group A,SerumNeopterin was significantly lower than baseline alter operation and IL-2 had no significant changes.In group H,both SerumNeopterin and IL-2 were significantly lower than baseline alter operation and 5 d after operation.Compared with group A,SerumNeopterin was significantly lower than baseline alter operation and 5 d after operation and IL-2 was significantly lower thanbaseline 5 d alter operation.Conchision:Autologous transfusion decreased the perioperative immune suppression in malignantstomach tumor patients.

Study on the correlation and predictive value of serum pregnancyassociated plasma protein A, triglyceride and serum 25-hydroxyvitamin D levels with gestational diabetes mellitus

BACKGROUND Gestational diabetes mellitus(GDM)is a concern due to its rapid increase in incidence in recent years.AIM To investigate the correlation and predictive value of serum pregnancyassociated plasma protein A(PAPP-A),triglyceride(TG),and 25-hydroxyvitamin D[25-(OH)D]with GDM in early pregnancy.METHODS A total of 99 patients in early pregnancy admitted to Peking University International Hospital from November 2015 to September 2017 were included,and underwent a fasting glucose test and oral glucose tolerance test screening at 24-28 wk of pregnancy.Of these cases with GDM,51 were assigned to group A and the remaining 48 cases without GDM were enrolled in group B.Serum PAPP-A,TG and 25-(OH)D in the two groups were compared and their correlation with blood sugar was analyzed.In addition,their diagnostic value in GDM was determined using receiver operating characteristic(ROC)curve analysis.RESULTS Group A had markedly lower serum PAPP-A and 25-(OH)D levels and a significantly higher serum TG level than group B,with statistical significance(P<0.05).Furthermore,Pearson analysis identified that PAPP-A and 25-(OH)D levels were negatively correlated with fasting blood glucose(FBG)levels(r=-0.605,P<0.001),(r=-0.597,P<0.001),while TG and FBG levels were positively correlated(r=0.628,P<0.001).The sensitivity,specificity,area under the curve(AUC)and optimal cut-off value of serum PAPP-A level in the diagnosis of GDM were 72.55%,82.35%,0.861 and 16.340,respectively,while the sensitivity of TG in diagnosing GDM was 86.27%,the specificity was 66.67%,the AUC was 0.813,with an optimal cut-off value of 1.796.The corresponding sensitivity,specificity,AUC and optimal cut-off value of serum 25-(OH)D were 64.71%,70.59%,0.721 and 23.140,respectively.Moreover,multivariate logistic regression analysis revealed that FBG,vascular endothelial growth factor,Flt-1,serum PAPP-A,TG,and 25-(OH)D were related risk factors leading to GDM in patients.CONCLUSION Serum PAPP-A,TG,and 25-(OH)D levels are all correlated with blood glucose changes in GDM,and are independent factors affecting the occurrence of GDM and have certain value in the diagnosis of GDM.

Downregulation of CD4+CD25+ regulatory T cells may underlie enhanced Th1 immunity caused by immunization with activated autologous T cells

Regulatory T cells （Treg） play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti-tumor immunity. We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti-tumor immunity and upregulated Thl responses in vivo. However, the underlying molecular mechanisms are not well understood. Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated autologous T cells. We found that Foxp3 expression decreased in CD4＋CD25＋ T cells from the immunized mice. Moreover, CD4＋CD25＋Foxp3＋ Treg obtained from immunized mice exhibited diminished immunosuppression ability compared to those from naive mice. Further analysis showed that the serum of immunized mice contains a high level ofanti-CD25 antibody （about 30 ng/ml, p〈0.01 vs controls）. Consistent with a role ofanti-CD25 response in the downregulation of Treg, adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice. The triggering of anti-CD25 response in immunized mice can be explained by the fact that CD25 was induced to a high level in the ConA activated autologous T cells used for immunization. Our results demonstrate for the first time that immunization with attenuated activated autologous T cells evokes anti-CD25 antibody production, which leads to impeded CD4＋CD25＋Foxp3＋ Treg expansion and function in vivo. We suggest that dampened Treg function likely contributes to enhanced Thl response in immunized mice and is at least part of the mechanism underlying the boosted anti-tumor immunity.

Serum outperforms plasma in small extracellular vesicle microRNA biomarker studies of adenocarcinoma of the esophagus

BACKGROUND Circulating microRNAs(miRNAs)are potential biomarkers for many diseases.However,they can originate from non-disease specific sources,such as blood cells,and compromise the investigations for miRNA biomarkers.While small extracellular vesicles(sEVs)have been suggested to provide a purer source of circulating miRNAs for biomarkers discovery,the most suitable blood sample for sEV miRNA biomarker studies has not been defined.AIM To compare the mi RNA profiles between matched serum and plasma s EV preparations to determine their suitability for biomarker studies.METHODS Matched serum and plasma samples were obtained from 10 healthy controls and10 patients with esophageal adenocarcinoma.s EV isolates were prepared from serum and plasma using Exo Quick TM and quantified using Nano Sight.RNA was extracted from s EV preparations with the mi RNeasy Serum/Plasma kit and profiled using the Taqman Openarray q PCR.The overall mi RNA content and theexpression of specific mi RNAs of reported vesicular and non-vesicular origins were compared between serum and plasma s EV preparations.The diagnostic performance of a previously identified multi-mi RNA biomarker panel for esophageal adenocarcinoma was also compared.RESULTS The overall mi RNA content was higher in plasma s EV preparations(480 mi RNAs)and contained 97.5%of the mi RNAs found in the serum s EV preparations(412 mi RNAs).The expression of commonly expressed mi RNAs was highly correlated(Spearman’s R=0.87,P<0.0001)between the plasma and serum s EV preparations,but was consistently higher in the plasma s EV preparations.Specific blood-cell mi RNAs(hsa-mi R-223-3 p,hsa-mi R-451 a,mi R-19 b-3 p,hsa-mi R-17-5 p,hsa-mi R-30 b-5 p,hsa-mi R-106 a-5 p,hsa-mi R-150-5 p and hsa-mi R-92 a-3 p)were expressed at 2.7 to 9.6 fold higher levels in the plasma s EV preparations compared to serum s EV preparations(P<0.05).In plasma s EV preparations,the percentage of protein-associated mi RNAs expressed at relatively higher levels(Ct 20-25)was greater than serum s EV preparations(50%vs 31%).While the percentage of vesicle-associated mi RNAs expressed at relatively higher levels was greater in the serum s EV preparations than plasma s EV preparations(70%vs 44%).A 5-mi RNA biomarker panel produced a higher cross validated accuracy for discriminating patients with esophageal adenocarcinoma from healthy controls using serum s EV preparations compared with plasma s EV preparations(AUROC 0.80 vs 0.54,P<0.05).CONCLUSION Although plasma s EV preparations contained more mi RNAs than serum s EV preparations,they also contained more mi RNAs from non-vesicle origins.Serum appears to be more suitable than plasma for s EV mi RNAs biomarkers studies.

Comparison of protein concentrations in serum versus plasma from Alzheimer’s patients

Background: There is great interest in developing blood-based biomarkers for Alzheimer’s disease (AD);however, there is no consensus as to what blood fraction is most appropriate for analyzing particular markers. The current study provides empirical evidence regarding how blood-based proteins vary depending on whether they are assayed in serum or plasma. Methods: Weanalyzed concentrations of 100 proteins in matched samples of serum and plasma from 39 Caucasian AD participants from the Texas Alzheimer’s Research and Care Consortium bymultiplex immunoassay. Results: Concentrations of 40 proteins were highly correlated (r2≥ 0.75) between plasma and serum while the remaining proteins were moderately to weakly correlated (r2< 0.75). Discussion: Whether plasma vs. serum is assayed can have a large impact on the observed concentration of some proteins, including several proteins that are of great interest to AD pathophysiology. The current findings may explain the significant discrepancies often times reported in the AD biomarker field.

Measurement of the Trace Elements Cu, Zn, Fe, and Mg and the Ultratrace Elements Cd, Co, Mn, and Pb in Limited Quantity Human Plasma and Serum Samples by Inductively Coupled Plasma-Mass Spectrometry

In public health studies limited volumes of blood are often collected and stored for future hypothesis testing. Archived samples are irreplaceable and therefore it is valuable to develop analytical techniques that require minimal sample vo-lume. This work describes the measurement of trace elements Mg, Cu, Fe, Zn and ultratrace elements Cd, Co, Mn, Pb in limited quantity (150 μL) human serum or plasma samples. Samples were digested using a hotblock and analyzed using inductively coupled plasma mass spectrometry (ICP-MS). The analytical method was evaluated using a quadrupole (Q) and sector field high resolution (SF) instrument to analyze trace elements in Seronorm? quality control serum material. The method was used to analyze 1888 blinded human plasma samples which were archived for the National Cancer Institute from the Nutrition Intervention Trial in Linxian China. The inductively coupled plasma method was capable of accurately analyzed limited quantity samples of human serum and plasma for the trace elements Mg, Cu, Fe Zn and the ultra trace elements Co, Mn and Pb. The concentration of Cd in human plasma samples was below the level of detection for 75% of the samples analyzed.

Effects of Storage Duration and Temperature of Human Plasma and Serum on Red Blood Cell Aggregation and Shape

Platelet-free plasma of human blood (sodium citrate and EDTA as an anticoagulant) and serum were stored at 4°C, room temperature (25°C) and at 37°C for 24 hours. RBC aggregation decreased after incubation of plasma and serum at 37°C for 4 hours. The RBC shape was changed at the same time: discocytes transformed to echinocytes. Storage of plasma and serum at 4°C and room temperature did not lead to significant alterations of RBC aggregation. The RBC shape did not change in influence of such plasma and serum. The most considerable decrease of RBC aggregation and change of their shapes were observed in the plasma and serum incubated at 37°C for 24 hours. Dilution of incubated plasma by fresh plasma led to consistent restoration of erythrocyte shape and their aggregation.

Indocyanine green plasma clearance rate and 99mTc-galactosyl human serum albumin single-photon emission computed tomography evaluated preoperative remnant liver

BACKGROUND Preoperative evaluation of future remnant liver reserves is important for safe hepatectomy.If the remnant is small,preoperative portal vein embolization(PVE)is useful.Liver volume analysis has been the primary method of preoperative evaluation,although functional examination may be more accurate.We have used the functional evaluation liver using the indocyanine green plasma clearance rate(KICG)and 99mTc-galactosyl human serum albumin single-photon emission computed tomography(99mTc-GSA SPECT)for safe hepatectomy.AIM To analyze the safety of our institution’s system for evaluating the remnant liver reserve.METHODS We retrospectively reviewed the records of 23 patients who underwent preoperative PVE.Two types of remnant liver KICG were defined as follows:Anatomical volume remnant KICG(a-rem-KICG),determined as the remnant liver anatomical volume rate×KICG;and functional volume remnant KICG(frem-KICG),determined as the remnant liver functional volume rate based on 99mTc-GSA SPECT×KICG.If either of the remnant liver KICGs were>0.05,a hepatectomy was performed.Perioperative factors were analyzed.We defined the marginal group as patients with a-rem-KICG of<0.05 and a f-rem-KICG of>0.05 and compared the postoperative outcomes between the marginal and not marginal(both a-rem-KICG and f-rem-KICG>0.05)groups.RESULTS All 23 patients underwent planned hepatectomies.Right hepatectomy,right trisectionectomy and left trisectionectomy were in 16,6 and 1 cases,respectively.The mean of blood loss and operative time were 576 mL and 474 min,respectively.The increased amount of frem-KICG was significantly larger than that of a-rem-KICG after PVE(0.034 vs 0.012,P=0.0273).The not marginal and marginal groups had 17(73.9%)and 6(26.1%)patients,respectively.The complications of Clavian-Dindo classification grade II or higher and post-hepatectomy liver failure were observed in six(26.1%)and one(grade A,4.3%)patient,respectively.The 90-d mortality was zero.The marginal group had no significant difference in postoperative outcomes(prothrombin time/international normalised ratio,total bilirubin,complication,post-hepatectomy liver failure,hospital stay,90-d,and mortality)compared with the not-marginal group.CONCLUSION Functional evaluation of the remnant liver enabled safe hepatectomy and may extend the indication for hepatectomy after PVE treatment.

Study of influence of sacubitril/valsartan on plasma NE, AngⅡ, ALD and serum sCD40L, sICAM-1, sFas, sFasL and cTnI, MMP-9 levels in patients with heart failure

Objective:To study the influence of sacubitril/valsartan on plasma NE, AngⅡ, ALD and serum sCD40L, sICAM-1, sFas, sFasL and cTnI, MMP-9 levels in patients with heart failure. Methods: To choose 42 cases of chronic heart failure in our hospital form August 2017 to December 2017 as the study group, and 42 cases of chronic heart failure form January 2017 to June 2017 as the control group. The patients in the control group were treated with 80 mg valsartan on the basis of general treatment, 1 times a day, and the patients in the study group were added with 50mg sacubitril/valsartan on the basis of general treatment, 2 times a day. To compare the plasma NE, AngⅡ, ALD and serum sCD40L, sICAM-1, sFas, sFasL and cTnI, MMP-9 levels before and after treatment in the two groups.Results:Before treatment, there was no statistical difference between the levels of plasma NE, AngⅡ, ALD and serum sCD40L, sICAM-1, sFas, sFasL and cTnI, MMP-9 in the two groups;(1) After treatment, the levels of plasma NE, AngⅡ, ALD in the control group compared with those before treatment in the seme group, there were significant differences, and there were significant differences between the two groups;(2) After treatment, the levels of serum sCD40L, sICAM-1, sFas, sFasL in the control group compared with those before treatment in the same group, there were significant differences, and there were significant differences between the two groups;(3) After treatment, the levels of serum cTnI, MMP-9 in the control group compared with those before treatment in the seme group, there were significant differences, and there were significant differences between the two groups.Conclusion: The application of sacubitril/valsartan to patients with heart failure on the basis of general treatment, not only could significantly improve the levels of plasma NE, AngⅡ, ALD, and also could significantly improve the levels of serum sCD40L, sICAM-1, sFas, sFasL and cTnI, MMP-9, the curative effect was more significant, it was worthy for clinical research and application.

MicroRNAs as potential biomarkers for diagnosis of post-traumatic stress disorder

Post-traumatic stress disorder is a mental disorder caused by exposure to severe traumatic life events.Currently,there are no validated biomarkers or laboratory tests that can distinguish between trauma survivors with and without post-traumatic stress disorder.In addition,the heterogeneity of clinical presentations of post-traumatic stress disorder and the overlap of symptoms with other conditions can lead to misdiagnosis and inappropriate treatment.Evidence suggests that this condition is a multisystem disorder that affects many biological systems,raising the possibility that peripheral markers of disease may be used to diagnose post-traumatic stress disorder.We performed a PubMed search for microRNAs(miRNAs)in post-traumatic stress disorder(PTSD)that could serve as diagnostic biomarkers and found 18 original research articles on studies performed with human patients and published January 2012 to December 2023.These included four studies with whole blood,seven with peripheral blood mononuclear cells,four with plasma extracellular vesicles/exosomes,and one with serum exosomes.One of these studies had also used whole plasma.Two studies were excluded as they did not involve microRNA biomarkers.Most of the studies had collected samples from adult male Veterans who had returned from deployment and been exposed to combat,and only two were from recently traumatized adult subjects.In measuring miRNA expression levels,many of the studies had used microarray miRNA analysis,miRNA Seq analysis,or NanoString panels.Only six studies had used real time polymerase chain reaction assay to determine/validate miRNA expression in PTSD subjects compared to controls.The miRNAs that were found/validated in these studies may be considered as potential candidate biomarkers for PTSD and include miR-3130-5p in whole blood;miR-193a-5p,-7113-5p,-125a,-181c,and-671-5p in peripheral blood mononuclear cells;miR-10b-5p,-203a-3p,-4488,-502-3p,-874-3p,-5100,and-7641 in plasma extracellular vesicles/exosomes;and miR-18a-3p and-7-1-5p in blood plasma.Several important limitations identified in the studies need to be taken into account in future studies.Further studies are warranted with war veterans and recently traumatized children,adolescents,and adults having PTSD and use of animal models subjected to various stressors and the effects of suppressing or overexpressing specific microRNAs.