Objective:To study the effects of brushite coating with strontium on the osteoblast-related factor expression of MC3T3-E1 cells.Methods:Brushite coating on tinanium surface was prepared by electrochemical deposition and the extract of the coating with stronium at 0.1%,0.5% and 1% were prepared respectively.MC3T3-E1 cells were cultured with the extracts,cell prolifera-tion,ALP activity,bFGF and VEGF mRNA were examined by MTT assay,ALP staining and RT-PCR respectively.Results:Stronti-um containing extracts promoted the proliferation,increased the ALP activity and mRNA expression of bFGF and VEGF(P <0.05)of MC3T3 cells.Conclusion:Brushite extract containing strontium may promote osteogenic function of MC3T3-E1 cells.

OBJECTIVETo investigate the application value of chemiluminescence method (CMIA) detection of Treponema pallidum (TP) specific antibodies in the blood test.

METHODSOver the same period the de novo enzyme linked immunosorbent assay (ELISA) and Abbott chemical luminescence method were used to detect the specific antibody of syphilis in a total of 66298 samples; TP-ELISA negative and TP-CMIA positive unpaid blood donation blood samples for syphilis specific antibody were detected and confirmed by Western blot.

RESULTSBlood samples from 66298 blood donors were detected by TP-ELISA, the positive samples was 250 and the positive rate was 0.38%. The positive samples of TP-CMIA was 297, the positive rate was 0.45%, the difference was statistically significant (P<0.05). The blood samples of 47 unpaid blood donors were confirmed by TP-Western blot method, as a result, 32 samples were positive, 15 were negative, and result detected by TP-ELISA method was negative.

CONCLUSIONTP-CMIA sensitivity is higher than that of TP-ELISA, and possesses higher sensitivity and specificity, and quick detection, simple operation, easy automation, suggesting greater application value in blood detection of Treponema pallidum.

OBJECTIVETo investigate the blood test patterns for blood donors after nucleic acid detection in blood center.

METHODSThe collected blood samples after voluntary blood donors first were detected by conventional ELISA, then 31981 negative samples were detected via HBV/HCV/HIV combined nucleic acid test of 6 mixed samples(22716 cases) or single samples(9265 cases) by means of Roche cobas s201 instrument. The combined detection method as follows: the blood samples were assayed by conventional nucleic acid test of 6 mixed samples, at same time, 6 mixed samples were treated with polyethylene glycol precipitation method to concentrate the virus, then the nucleic acid test of blood samples was performed; the single detection method as follows: firstly the conventional nucleic acid test of single sample was performed, then the positive reactive samples after re-examination were 6-fold diluted to simulate the nucleic acid test of 6-mixed samples. The positive rate of positive samples detected by combined nucleic acid test, positive samples detected by nucleic acid test of mixed virus concentration and positive samples detected by single nucleic acid test was statistically analyzed. In addition, for HBVpersons the serological test yet should be performed.

RESULTSIn 22 716 samples detected by nucleic acid test of 6 mixed samples (MP-6-NAT) , 9 cases were HBV(0.40‰, 9/22716); at same time, the detection of same samples by nucleic acid test of mixed sample virus concentration showed 29 cases of HBV(1.28‰, 29/22716). In 9265 samples detected by single nucleic acid test(ID-NAT) 12 cases showed HBV(1.30‰, 12/9265), meanwhile the detection of these 12 samples with HBVby 6-fold dilution for virus concentration found only 4 samples with HBV. In serological qualified samples, ID-NAT unqualified rate was 1.28‰, which was higher than that of MP-6-NAT(0.4‰) (χ=8.11, P<0.05); but there was no statistical difference between unqualified rate of ID-NAT and MP-6-NAT(1.3‰ vs 1.28‰)(χ=0.00, P>0.05). In 41 samples with HBsAgHBV DNAdetected by ELISA, 36 samples were confirmed to be occult HBV infective(OBI) by HBsAb, HBcAb test of ELISA; out of these 41 samples, 33 samples showed HBcAb(91.66% of OBI), 5 might be HBV "window period" infective, moreover the HCV RNA and HIV RNA positive samples were not found.

CONCLUSIONTo avoid the missdiagnosis of donors with low level of virus, the nucleic acid test must be carried out after virus concentration of mixed samples when the blood test pattern of donors is nucleic acid test of mixed samples, otherwise the single nucleic acid test must be performed to obtain more high detected rate of virus nucleic acid. The HBcAb serologic test and physical examination of donors before blood donation must be enhanced on basis of serological test of HBsAg; for high risk people, the persuading no blood donation is simplest pattern.

OBJECTIVETo explore the key technique for preparation of the frozen platelet and efficacy of its clinical application.

METHODSThe influences of the donators' peripheral platelet count, starting time of freeze, injection rate and evenness of the freeze-protective agent, storage mode, re-melting temperature and the capacity of water-bath etc. on the quality of the frozen platelets were analyzed retrospectively in 3 257 samples of frozen platelets before platelet pheresis. Then, the platelet counts were examined in 150 cases transfused with frozen platelets at the time-points of 1, 24, 48 and 72 hrs after transfusion, 90 cases suffered from the obstetrical bleeding were transfused with 200 parts of the re-melting frozen platelets, and then the peripheral blood platelet count, platelet increasing index(CCI), bleeding time and blood clot retraction rate etc. were observed for determining the clinical efficiency of the frozen platelets.

RESULTSThe floccule in the re-melting frozen platelets from the donators with (175-250)×10(9)/L platelets were decreased significantly(P<0.01). The quality of frozen platelets was influenced by the following factors, such as injection of DMSO at a too fast and heterogeneous rate, blood bags stored in a multilamminar space, and re-melting in a water-bath of small capacity etc. The routine storage for 0 and 3 days did not influence the quality of the frozen platelets. The recovery rate of one year-freezing platelets all was higher than 80%. The effects of the frozen platelets transfused into the patients with obstetrical bleeding displayed good haemostatic results, and the blood transfusion reaction did not occur. However, the frozen platelets immediately were exhausted and displayed their function, but the counting after 48 hrs could not display a good effect of raising platelet number.

CONCLUSIONSThe peripheral platelet count before platelet pheresis, the injection rate and evenness of the protective agent, the number of stratum for blood bags and the capacity of re-melting water-bath etc. all are the key factors influencing the quality of the frozen platelets. The frozen platelets prepared in this study shows a good efficacy of clinical application.

Blood Platelets ; Blood Preservation ; Blood Transfusion ; Freezing ; Hemostasis ; Humans ; Platelet Count ; Platelet Transfusion ; Plateletpheresis ; Transfusion Reaction

The purpose of this study was to detect the distribution of Treg and Th17 cells in bone marrow and to investigate the relationship of Treg/Th17 imbalance with the pathogenesis and progression of multiple myeloma (MM). The Bone marrow was collected from 37 MM patients and 12 healthy volunteers, the ratio of Treg and Th17 cells was detected by flow cytometry. The expression of Treg and Th17 cells simultaneously was examined in peripheral blood of 19 MM patients with same method. The results indicated that the frequency of Treg cells was higher in MM patients than that in control group (P < 0.05), there was a trend of increasing of Treg cell number in the ISS stage from I+II to III (P < 0.05). Furthermore, in the patients with MM, the Treg cell number in bone marrow was higher than that in peripheral blood (P < 0.05). Th17 cell rate was not statistically different between MM patients and control group (P > 0.05), and at different ISS stage (P > 0.05). Th17 cell number between bone marrow and peripheral blood was not significantly different (P > 0.05).The ratio of Treg/Th17 in patients with MM was higher than that in control group (P < 0.05), and increased gradually from ISS stage I+II to stage III (P < 0.05). It is concluded that the Treg/Th17 immune imbalance is presenced in bone marrow of patients with MM, this imbalance may promote the progression of MM.
Bone Marrow ; immunology ; Cell Count ; Disease Progression ; Flow Cytometry ; Humans ; Multiple Myeloma ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

OBJECTIVETo investigate the changes of thrombospondin 1(TSP1) level and von Willebrand factor cleaving protease(ADAMTS13) activity in the patients with hematologic malignancies before and after treatment and to evaluate their clinical significance.

METHODSEighty-two patients with hematologic malignancies were enrolled in this study, among them 20 patients were with acute leukemia, 48 patients were with lymphoma and 14 patients were with multiple myeloma. The plasma samples of 82 patients with hematologic malignancies and 45 healthy controls were collected. The activities of ADAMTS13 were evaluated by residue collagen binding assay(R-CBA), the levels of TSP1 and vWF antigen were measured by enzyme-linked immunosorbent assay(ELISA).

RESULTSThe activity of plasma ADAMTS13 in patients with hematologic malignancies was lower than that of normal controls(P<0.05). The levels of vWF antigen and TSP1 in the patients with hematologic malignancies were higher than those in normal controls(P<0.05). After standard induction chemotherapy, the ADAMTS13 activity of the patients with hematologic malignancies at the complete remission was higher than that before therapy(P<0.05); the vWF antigen level was significantly lower than that in the patients with hematologic malignancies before therapy(P<0.05), but still higher than that in controls(P<0.05). There were 25 infection patients in 82 cases of hematologic malignancies, and the ADAMTS13 activity in the patients with newly diagnosed hematologic malignancies complicated with infection before therapy was obviously lower than that in the patients with hematologic malignancies without infection(P<0.05), the levels of vWF antigen and TSP1 were significantly lower than that in patients without infection (P<0.05). In the process of treatment, 8 patients have been speculated to suffer from thrombus, and the ADAMTS13 activity in the patients with thrombus was obviously lower than that in the patients without thrombus(P<0.05).

CONCLUSIONLow ADAMTS13 activity and high TSP1 level may participate in the progress of hematologic malignancies, the infection and thrombotic events may lead to further reduction of the ADAMTS13 activity. Assaying the level of ADAMTS13 activity in the patients with malignant tumor may be helpful to prevent the infection and thrombosis in the patients with hematologic malignancies.

OBJECTIVETo detect the plasma activity of von Willebrand factor-cleaving protease (ADAMTS13) in the patients with prothrombotic status, and explore the effect and significance of ADAMTS13 in the prothrombotic status. The correlation of ADAMTS13 with von Willebrand factor (vWF), thrombospondin 1 (TSP1), C-reactive protein etc, and blood pressure was simultaneously analyzed．

METHODSThe activity of ADAMTS13 in patient groups (atherosclerosis, diabetes, acute promyelocytic leukemia, cancer and sepsis, a total of 260 cases) and in control group 50 cases were evaluated by residue collagen binding assay（R-CBA）, the protein levels of TSP1 and vWF were measured by ELISA kits； the correlation of ADAMTS13 activity with CRP, creatinine, and blood pressure was analyzed with statistical soft ware.

RESULTSThe activity of plasma ADAMTS13 in patient group was significantly lower than that in normal control group(P<0.05). And the protein levels of TSP1 and vWF in the patients with prothrombotic status were higher than those in the normal controls(P<0.05). Analysis of the correlation showed that the ADAMTS13 activity correlated negatively with the levels of TSP1 protein, blood sugar, blood pressure, D-dimer, creatinine,and CRP levels (P<0.05), however, the ADAMTS13 activity did not significantly correlate with the levels of serum lipids, blood type, platelet number and hemoglobin level(P>0.05).

CONCLUSIONThe plasma ADAMTS13 activity is decreased in the patients with prothrombotic status, suggesting that the decreased ADAMTS13 activity may participate in the occurrence of prothrombotic status, and the dectection of plasma ADAMTS13 activity may help the diagnosis of pro-thrombotic disease.

ADAMTS13 Protein ; Factor Analysis, Statistical ; Fibrin Fibrinogen Degradation Products ; Humans ; Sepsis ; Thrombosis ; von Willebrand Factor

OBJECTIVETo investigate the clinical efficacy and immune mechanism of immunotherapy of dendritic cells (DC) and cytokine-induced killer cell (CIK) combined with chemotherapy in patients with newly diagnosed multiple myeloma (MM).

METHODStwenty-two newly diagnosed MM patients were chosen and divided into two groups, out of them,12 patients in single chemotherapy group were treated by chemotherapy only, 10 patients in combined group were treated by adoptive immunotherapy (DC-CIK) combined with chemotherapy. Using flow cytometry, the CD4 Treg cells in the peripheral blood of 22 MM patients were detected before and after treatment. And the clinical outcomes between two groups were also compared.

RESULTSAfter treatment the overall response rate(ORR) of patients in the single chemotherapy group was 50% (6/12), among them 2 cases were in complete remission (CR) (16.67%), 2 cases very good partial remission (VGPR) (16.67%), 2 cases were in partial remission (PR) (16.67%). However, the ORR of patients in immunotherapy combined with chemotherapy group was 70.% (7/10), including in 3 cases of CR (30%), 2 cases of VGPR (20%), 2 cases of PR (20%). Compared to healthy volunteers, the proportion of Treg cells in peripheral blood of two groups before treatment was significantly higher (P<0.05). In contrast, the proportion of Treg cells in the peripheral blood of above-mentioned 2 groups after treatment was reduced significantly (P<0.05). In addition, compared to chemotherapy group, the proportion of Treg cells in the combined group decreased significantly (P<0.05). The further analysis found that the proportion of Treg cells in the peripheral blood of the 2 groups was not significant changed (P>0.05) in the patients with ineffictive clinical treatment, but the proportion of Treg cells significantly decreased (P<0.05) in the patients with effective clinical treatment.

CONCLUSIONDC-CIK immunotherapy can synergize or enhance the effect of chemotherapeutics, alleviate the immune dysfunction in MM; and DC-CIK immunotherapy combined with chemotherapy can elevate the clinical efficacy in patients with newly diagnosed multiple myeloma.

Antineoplastic Agents ; Cytokine-Induced Killer Cells ; Dendritic Cells ; Flow Cytometry ; Humans ; Immunotherapy ; Multiple Myeloma ; Remission Induction ; T-Lymphocytes, Regulatory ; Treatment Outcome

OBJECTIVETo analyze in vitro the effect of mesenchymal stem cells (MSCs) on secreting cytokines by T lymphocytes and ratio of CD4⁺CD25⁺ T cells from patients with immune thrombocytopenia (ITP).

METHODSHuman bone marrow-derived MSCs were isolated by Ficoll Hypaque and cultured for proliferating to passage cells. Allogeneic T lymphocytes of health adults and ITP patients were isolated from peripheral blood by Ficoll Hypaque and nylon cotton column, and the ratio of CD4⁺CD25⁺ T cells was detected by flow cytometry. Then the different amounts of 1 × 10⁴, 5 × 10⁴, 2 × 10⁵ MSCs per well treated with mitomycin as stromal feeder layers were co-cultured with above-mentioned T lymphocytes, 5 days after cocultivation, the ratio of CD4⁺CD25⁺ T cells was detected by flow cytometry and the levels of IL-2, IFN-γ, IL-4, IL-10 were measured by enzyme- linked immune sorbent assay (ELISA).

RESULTSAfter co-cultured with 2 × 10⁵ MSCs for 5 days, the ratio of CD4⁺CD25⁺ T cells and CD4⁺CD25⁺/CD4⁺ were significantly higher than of separate T lymphocytes in ITP patients [(4.56 ± 0.70)% vs (2.24 ± 0.81)%, (9.91 ± 1.18)% vs (4.08 ± 1.17)%, respectively] (P<0.05). To compare with separate T lymphocytes in ITP patients, the cytokine concentrations of IL-2 and IFN-γ from the culture supernatants significantly reduced from (280.47 ± 17.33) pg/ml to (97.21 ± 12.07) pg/ml and from (129.33 ± 16.34) pg/ml to (72.75 ± 7.81) pg/ml, respectively. In contrast, the cytokine concentrations of IL-4 and IL-10 increased from (16.34 ± 2.60) pg/ml to (37.98 ± 4.05) pg/ml and from (54.78 ± 5.62) pg/ml to (113.77 ± 5.68) pg/ml, respectively.

CONCLUSIONMSCs significantly inhibited the cytokine levels of IL-2 and IFN-γ secreted by Th1 cells and promoted the releases of IL-4 and IL-10 by Th2 cells in ITP , thereby regulating the balance between Th1 and Th2 reaction, as well as up-regulating the expression of CD4⁺CD25⁺ T cells in vitro,then induced the immunologic tolerance of ITP.

Adolescent ; Adult ; CD4-Positive T-Lymphocytes ; secretion ; Cells, Cultured ; Female ; Flow Cytometry ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; Middle Aged ; Thrombocytopenia ; metabolism ; Young Adult

Objective: To analyze the epidemiological characteristics of acute paraquat(PQ)poisoning in children in southwest Shandong, and the risk factors for pulmonary interstitial fibrosis. Methods: This retrospective study was performed on the clinical data of children with acute PQ poisoning admitted from January 2013 to December 2017 in 12 hospitals in southwest Shandong.All participants were divided into pulmonary interstitial fibrosis group and no pulmonary interstitial fibrosis group on the basis of the chest CT 14 days after poisoning.The epidemiological characteristics and risk factors of pulmonary interstitial fibrosis were analyzed. Results: During the study period, a total of 307 children with acute PQ poisoning were admitted to 12 hospitals, of which 61 (19.87%) were suffering from acute PQ poisoning.Forty-nine cases with complete clinical data were analyzed, including 26 male and 23 female patients poisoned by oral.The age distribution ranged from 8 months to 14 years.Poisoning mainly occured from July to September of each year.The mortality of acute PQ poisoning was 8.2%(4/49), and the incidence of pulmonary interstitial fibrosis in survival patients was 44.4%(20/45). Statistical differences (P<0.05) were found between the pulmonary interstitial fibrosis and no pulmonary interstitial fibrosis, with regard to the times of blood purification, the time from poison exposure to blood purification, the application rate of glucocorticoids, the concentration of PQ in urine, the pediatric critical illness score, the time from poison exposure to gastric lavage, the white blood count at admission, serum creatinine, arterial blood lactate, PaO₂, PaCO₂, and PaO₂/FiO₂; however, there was no significant difference in the proportion of blood purification treatment, the mode of blood purification treatment, alanine aminotransferase, aspartate aminotransferase, urea nitrogen, creatine kinase and troponin.Stepwise logistic regression analysis showed that the time from exposure to poison to gastric lavage(OR=0.683, 95%CI 0.210-2.222)and to blood purification(OR=0.0133, 95%CI 0.004-0.042), the times of blood purification(OR=2.862, 95%CI 1.450-5.648), concentration of PQ in urine(OR=1.435, 95%CI 1.085-1.898), and the use of glucocorticoids(OR=0.190, 95%CI 0.048-0.757) were the risk factors for pulmonary interstitial fibrosis(P<0.05). Conclusion Early gastric lavage and blood purification, increasing the frequence of adminitrating purification appropriately, using low-dose glucocorticoids can reduce the incidence of pulmonary interstitial fibrosis of children with acute PQ poisoning.