This study was purposed to investigate the feasibility of cryopreservation platelets under -80 degrees C after stored for 3 days at normal temperature and its clinical use. The platelet count, aggregation functions, adhesive ability, hypotonic shock reaction and CD62p expression of platelets preserved at normal temperature, cryopreserved on same day of collection and after storage for 3 days were detected, and possibility of using platelets cryopreserved after storage for 3 days in clinic was evaluated by comparison of contactable clinical cases. The results indicated that no significant differences in platelet count, hypotonic shock reaction and adhesive ability were found within 3 days of storage (p > 0.05); but differences in aggregation function and CD62p expression were significant (p < 0.05). As compared with platelets cryopreserved on same day of collection, the detected parameters such as platelet count, aggregation function, adhesive ability, hypotonic shock reaction, expression at storage for 3 days did not show significant differences (p > 0.015; CCI value in clinically use of them also did not show significant difference too (p > 0.05). It is concluded that the platelets after storage for 3 days can be cryopreserved, the efficiency of them in clinical use is not significant difference from platelets cryopreserved on same day of collection.
Adult ; Blood Platelets ; Blood Preservation ; methods ; Cryopreservation ; methods ; Humans ; Platelet Aggregation ; Platelet Count ; Young Adult

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

OBJECTIVE: To study the association between superior mesenteric artery hemodynamic indexes and scores of lower gastrointestinal symptoms rating scales(LGSRS) in patients with type 2 diabetic mellitus. METHODS: Totally 142 inpatients with type 2 diabetes with average age of 58.76±12.32 yrs were enrolled, who were treated from August 2016 to March 2018. The history, gender, age,course and BMI were recorded, and fasting blood glucose(FBG), glycosylated hemoglobin(HbA1c), 2-hour postprandial blood glucose(PBG), total cholesterol(TC), triglyceride(TG), urine ACR and LGSRS were determined. Ultrasonic scanning of mesenteric artery was performed for hemodynamic indexes, including artery inner diameter(ID), peak systolic velocity(PSV), end-diastolic velocity(EDV), and resistance index(RI)at starting part,first level branch, and second level branch from root of the superior mesenteric artery(SMA).Patients were divided into 2 groups according to their LGSRS, 74 patients with LGSRS≥6 were in positive group, and 68 patients with LGSRS<6 were in control group. RESULTS: 1. There were no significant differences between the 2 groups in gender, BMI, HbA1C, FBG, PBG,TC or TG(P>0.05), but the age and DD were significantly higher in positive group than in control group(P<0.05). 2. There were three patients(4.41%) who had atheroma or plaques in SMA in control group, while five patients in positive group(6.76%)(P>0.05). 3. There were no significant difference between positive group and control group in ID at starting part and first level branch of SMA, while ID at second level branch was significantly increased in positive group compared with control group [(3.83±0.85)mm vs.(3.53±0.90)mm, P<0.05)].4. RI at first(0.816±0.059 vs 0.842±0.063,P<0.05) and second level branch(0.813±0.076 vs 0.845±0.073, P<0.05) and PSV at first level branch[(110.89±46.89)cm/s vs(95.72±36.59)cm/s,P<0.05] were significantly high in positive group; there were no difference in other hemodynamic indexes between the groups. 5.Adjusted by age,DD,glycemic and lipidemic profile,Logistic regression showed that ID at first(RR=2.092,95%CI 1.080-4.050,P=0.029) and second level branch(RR=0.491,95%CI 0.252-0.955,P=0.36) and EDV at second level branch(RR=0.897,95%CI 0.824-0.976,P=0.012) were independent factors influencing LGSRS(P<0.05). CONCLUSION: Ultrosonic hemodynamic abnormalities in the superior mesenteric artery might be important factor in development of lower gastrointestinal tract symptoms in patients with type 2 diabetes.

Purpose To investigate the expression of the protein of CD151 and MT1-MMP in adenocarcinoma of esophagogastric junc-tion tissues and to explore their relationship with the invasiveness and metastasis of the tumor. Methods CD151 and MT1-MMP pro-tein were detected by immunohistochemical staining respectively in 15 cases of paraneoplastic normal gastric mucosa tissue, 40 cases dysplasia and 172 cases of adenocarcinoma of esophagogastric junction tissues. Results All of the expression level of CD151 and MT1-MMP protein were increasing according to the order of normal-dysplsia-carcinoma. The expression rate of CD151 was 6. 67%, 20. 0% and 78. 3%, while the rate of MT1-MMP with 13. 3%, 22. 5% and 79. 1% in the normal, dysplasia and carcinoma subgroup. The expression rate of each protein were significant difference among the three goroups (P<0. 05). In the adenocarcinoma of esopha-gogastric junction tissues, the expression of them in the subgroup of poorly-differentiated with serosa invasion and lymph nodes metasta-sis was significantly higher than the other subgroup of well-differentiated with non serosa invasion or lymph nodes metastasis ( P <0. 05 ) . There was a positive correlation between the expression of CD151 and MT1-MMP protein in adenocarcinoma of esophagogastric junction tissues ( P<0. 05 ) . Conclusions CD151 and MT1-MMP assuredly and highly express in the adenocarcinoma of esopha-gogastric junction tissues and they have close relationships, which could involved in the invasiveness and metastasis synergistically in this tumor.

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 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 efficacy of dendritic cells and cytokine-induced killer cells (DC-CIK) combined with chemotherapy for treating newly diagnosed patients with multiple myeloma (MM) and their effect on cellular immune functions of CD4(+) CD25(+) Treg cells in peripheral blood after adoptive immunotherapy.

METHODSFouty two patients with MM were randomly divided into two groups: chemotherapy group and combined therapy group; 20 patients in chemotherapy group were treated by chemotherapy only, 22 patients in combined therapy group were treated by adoptive immunotherapy (DC-CIK) combined with chemotherapy, and the clinical outcomes of patients and the levels of CD4(+) CD25(+) Treg cells in peripheral blood between 2 groups were compared.

RESULTSAfter treating for 3 weeks, the quality of life, clinical index and survival of patients in combined therapy group were better than those of patients in chemotherapy group (P < 0.05); the ratios of CD4(+) CD25(+)/CD4(+) and CD4(+) CD25(+) FoxP3(+)/CD4(+) CD25(+) of patients in combined therapy group were obviously lower than those of patients in chemotherapy group (P < 0.05).

CONCLUSIONThe immunotherapy of DC-CIK can strengthen the activities of CD4(+) CD25(+) Treg cells, which combined with chemotherapy can be an effective and promising effects for treatment of patients with MM.

Cell- and Tissue-Based Therapy ; Cytokine-Induced Killer Cells ; cytology ; Dendritic Cells ; cytology ; Humans ; Immunotherapy, Adoptive ; Multiple Myeloma ; drug therapy ; therapy ; T-Lymphocytes, Regulatory ; cytology ; Treatment Outcome

OBJECTIVETo evaluate the treatment value of adoptive immunotherapy (dendritic cells and cytokine-induced killer cells, DC-CIK) combined with chemotherapy on patients with multiple myeloma (MM) and its effect on secreting function of T lymphocytes in MM patients.

METHODSA total of 36 patients with MM were randomly divided into two groups, among them 28 patients in chemotherapy group were treated by chemotherapy only, 28 patients in combined therapy group were treated by adoptive immunotherapy (DC-CIK) combined with chemotherapy, and the clinical outcomes and the levels of IL-2, IFN-γ, IL-4, IL-10 secreted by T lymphocytes between two groups were compared.

RESULTSAfter treatment, the quality of life, clinical index and survival in combined therapy group were better than those in chemotherapy group (P <0.05); the levels of IL-2 and IFN-γ in combined therapy group was higher than these in chemotherapy group (P <0.05), and the levels of IL-4 and IL-10 in combined therapy group were lower than those in chemotherapy group (P <0.05).

CONCLUSIONDC-CIK combined with chemotherapy can be an effective and promising treatment for patients with MM, and it maybe strengthen the anti-tumor action of bodies by regulating the balance between Th1 and Th2 reaction.

Cytokine-Induced Killer Cells ; Dendritic Cells ; Humans ; Immunotherapy ; Immunotherapy, Adoptive ; Interleukin-10 ; Interleukin-2 ; Interleukin-4 ; Multiple Myeloma ; Quality of Life ; T-Lymphocytes

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

OBJECTIVE: To explore the effect of miR-144 to the biological behavior of multiple myeloma cells and its mechanism. METHODS: RT-PCR was used to detect the expression of miR-144 in multiple myeloma cells and plasma of MM patients. MTT assay was used to detect the proliferation and cloning ability of myeloma cells transfected by miR-144. Flow cytometry was used to detect the cell cycle distribution of myeloma cells with over-expression of miR-144. Apoptosis of myeloma cells with over-expression of miR-144 was detected by TUNEL assay. Transwell cell invasion and migration assay was used to detect the invasion and migration ability of myeloma cells with overexpressing on miR-144.Western blot analysis was used to detect the protein expression levels of MMP-9 and MMP-2 in myeloma cells with over expression of miR-144, as well as the expression levels of proteins related to Wnt/β-catenin signaling pathway. RESULTS: The expression level of miR-144 in MM cell lines and blood of MM patients was significantly lower than that in control group (P＜0.05). The proliferation, invasion and migration of myeloma cells with over-expression of miR-144 were significantly decreased (P＜0.05), and the apoptosis level was increased (P＜0.05). The expression levels of MMP-9, MMP-2, Wnt/β-catenin signaling pathway in myeloma cells with over-expression of miR-144 were significantly lower than those in control group (P＜0.05). CONCLUSION MiR-144 can inhibit the proliferation, migration and invasion of multiple myeloma cells and induce cell apoptosis. The specific mechanism may be related with the activity of inhibiting Wnt/β-catenin signaling pathway.
Apoptosis ; Biological Products ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; Multiple Myeloma ; Wnt Signaling Pathway ; Wnt4 Protein ; beta Catenin