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【Objective】 To analyze the efficacy of ABO-matched platelet transfusions and ABO-mismatched platelet transfusions in patients with hematonosis and to explore the effect of circulating immune complexes (CIC) on the efficacy. 【Methods】 A total of 1 510 platelet transfusions involving 757 patients in our hospital from January 2013 to June 2018 were retrospectively analyzed. The patients were divided into ABO-matched group and ABO-mismatched group. The 12-hour percent platelet recovery (PPR) was used to evaluate the effect of platelet transfusion between the groups. TEG was used to evaluate the efficacy of the transfusions, and CIC value was measured before and after platelet transfusion. The effect of A-B/CIC (or AB-O/CIC) on platelet function was tested. 【Results】 1)The results showed that platelet transfusion was effective(PPR>30%) in both ABO-matched group[PPR=(66.5±52.8)%] and ABO-mismatched group[PPR=(47.7%±51.6)%], and there was no increase in the report of hemolytic transfusion reaction of ABO-mismatched group. The efficacy of ABO-matched platelet transfusions was significantly better than that of ABO-mismatched group(P 0.05. 2) In the experiment of simulating platelet transfusion in patients, no difference in MA value of TEG was noticed between ABO-mismatched groups and ABO-matched groups (all P>0.05). 3) There was no difference in CIC value before and after platelet transfusions (P>0.05) in the ABO-matched group, while CIC value decreased significantly in all ABO-mismatched groups (all P < 0.05). 4) The MA values (mm)of AB, A and O blood group platelets mixed with A-B/CIC and AB-O/CIC were 36.1 vs 31.1, 37.8 vs 35.0 and 43.1 vs 45.7, with the MA value (mm) in control group at 49.2 vs 49.5, respectively. 【Conclusion】 Platelet transfusion was effective in both ABO-matched group and ABO-mismatched group, and the efficacy of ABO-matched group was significantly better compared with the ABO-mismatched group. There was no increase in the safety risk of ABO-mismatched platelet transfusion with major mismatches/minor matches. CIC can inhibit the function of platelets and combine more with ABO-matched platelets than with ABO-mismatched platelets, therefore, CCI is an important influencing factor on the efficacy of platelet transfusions.
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BACKGROUND:The Wnt/β-catenin signaling pathway is one of the most important signaling pathways in stem cel regulation, which is involved in regulation of cel proliferation and differentiation. OBJECTIVE:To investigate the expression of Wnt/β-catenin main signaling molecule in inflammatory bowel tissues treated with bone marrow mesenchymal stem cel transplantation. METHODS:2,4,6-Trinitrobenzene sulfonic acid was used for establishing inflammatory bowel diseases rat models. Bone marrow mesenchymal stem cels labeled with green fluorescent protein were transplanted into rat modelsviatail vein. Normal saline was injected as control. The expression of Wnt/β-catenin signaling molecule was detected in the large intestine tissue of inflammatory bowel disease rat models by quantitative RT-PCR at 14 and 28 days after transplantation. RESULTS AND CONCLUSION:Real-time quantitative PCR results showed that the expression of Wnt3a andβ-catenin in the inflammatory bowel tissue increased significantly (P 0.05). The expressions of Wnt3a, β-catenin and c-myc in the transplantation group were significantly lower than those in the control group after transplantation (P <0.05). These findings indicate that the Wnt/β-catenin signaling pathway plays important roles in inflammatory bowel disease and repair after bone marrow mesenchymal stem cel transplantation, while this pathway may promote stem cels differentiating into intestinal epithelium, promote recovery from inflammatory bowel disease, repair inflammatory area, and restore intestinal tissue homeostasis.
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<p><b>OBJECTIVE</b>To explore the influence of exogenous putrescine on the function of liver and apoptosis of liver cells in rats.</p><p><b>METHODS</b>Ninety healthy clean SD rats were divided into control group (C, n = 10, intraperitoneally injected with 2 mL normal saline), low dosage putrescine group (LP, n = 40), and high dosage putrescine group (HP, n = 40) according to the random number table. Rats in the latter two groups were intraperitoneally injected with approximately 2 mL putrescine (2.5 or 5.0 g/L) with the dosage of 25 or 50 µg/g. Ten rats from group C at post injection hour (PIH) 24 and 10 rats from each of the latter two groups at PIH 24, 48, 72, 96 were sacrificed. Heart blood was obtained for determination of serum contents of ALT and AST. Liver was harvested for gross observation and histomorphological observation with HE staining. Apoptosis was shown with in situ end labeling, and apoptosis index (AI) was calculated. Data among the three groups and those at different time points within one group were processed with one-way analysis of variance or Welch test; LSD or Dunnett's T3 test was used for paired comparison; factorial design analysis of variance of two factors was applied for data between group LP and group HP.</p><p><b>RESULTS</b>(1) No obvious abnormality was observed at gross observation of liver of rats in each group. Liver tissue of rats in group C was normal. Light edema was observed occasionally in liver of rats in groups LP and HP, but necrotic cells were not seen. (2) Content of ALT at PIH 24, 48, 96 and content of AST at PIH 72 and 96 in group LP were respectively (38 ± 10), (45 ± 6), (34 ± 4), (207 ± 18), (196 ± 19) U/L, and content of ALT at PIH 72 and 96 and content of AST at PIH 24, 72, 96 in group HP were respectively (38 ± 6), (48 ± 5), (213 ± 43), (209 ± 40), (230 ± 29) U/L. They were significantly higher than those of rats in group C [(29 ± 5), (163 ± 42) U/L, with P values all below 0.01]. There were statistically significant differences between group LP and group HP in the content of ALT at PIH 48, 72, 96 and content of AST at PIH 96 (with P values all below 0.05). Compared with that at PIH 24 of each group, content of ALT of rats in group LP at PIH 48 and that of rats in group HP at PIH 96, as well as content of AST of rats in group LP at PIH 48, 72, 96 and that of rats in group HP at PIH 48 were significantly increased or decreased (with P values all below 0.05). Factorial analysis showed that the differences due to different concentration of putrescine on content of AST were statistically significant (F = 12.21, P = 0.001), but not on content of ALT (F = 0.01, P = 0.974) between group LP and group HP. (3) AI values of rats in group LP at PIH 24, 48, 72 were respectively (5.69 ± 0.38)%, (13.80 ± 1.66)%, (11.56 ± 1.74)%, and AI values of rats in group HP at PIH 72 and 96 were respectively (10.29 ± 1.43)%, (15.29 ± 1.41)%. They were all obviously higher than AI value of control group at PIH 24 [(3.50 ± 0.30)%, with P values all below 0.01]. There were statistically significant differences between group LP and group HP in AI value at PIH 24, 48, 96 (with P values all below 0.05). Compared with that at PIH 24 of each group, AI value of rats in groups LP and HP at PIH 48, 72, 96 were significantly increased or decreased (with P values all below 0.05). Factorial analysis showed that the differences in the influence of concentration of putrescine and stimulation time on AI value were statistically significant (with F values respectively 22.95 and 130.44, P values all below 0.01).</p><p><b>CONCLUSIONS</b>Intraperitoneal injection of exogenous putrescine in the dosage of 25 or 50 µg/g could lead to certain degree of functional damage of liver and apoptosis of liver cells of rat. The higher the dosage and the longer the stimulation time, the more obvious the damage and apoptosis would be.</p>
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Animals , Rats , Alanine Transaminase , Blood , Apoptosis , Hepatocytes , Cell Biology , Liver , Cell Biology , Pathology , Putrescine , Toxicity , Rats, Sprague-DawleyABSTRACT
BACKGROUND:Previous studies have verified that mesenchymal stem cells could be transplanted into inflammatory bowel mucosa to repair inflammatory bowel tissue. OBJECTIVE:To observe the differential gene expression in large intestine before and after mesenchymal stem celltransplantation in repair of inflammatory bowel tissue of rats using microarray technology, and to primarily discover the main genes during mesenchymal stem celltransplantation, differentiation, and reparation in inflammatory colorectal tissue region. METHODS:Healthy Sprague-Dawley rats were randomly divided into two groups. Experimental rat models of inflammatory bowel disease were established using trinitrobenzene sulfonic acid via enema. At 24 hours after model establishment, green fluorescent protein-labeled mesenchymal stem cells were infused via the caudal vein. The control group was treated with physiological saline by enema, instead of trinitrobenzene sulfonic acid. At 28 days, large intestine was obtained from the experimental group and control group. Differential y expressed genes were screened in the experimental and control groups using microarray technique. RESULTS AND CONCLUSION:The microarray analysis results showed that there were 388 differential genes in the control and experimental groups (P2), in which 191 were up-expressed, and 197 were down-expressed. Al of these genes were mainly involved in inflammatory reaction, immune reaction and celldifferentiation. In the top 10 up-regulation and down-regulation differential genes (total y 20 genes), 3 genes were involved in inflammation, 3 genes were involved in immune reaction, and 2 genes were related to stem celldifferentiation. In the 388 genes, 33 were related to signaling pathways (P<0.05), 6 related to inflammation, 8 related to immunity, and 5 related to stem celldifferentiation. Results suggested that the main genes involved in mesenchymal stem cells in repair of inflammatory bowel tissue were primarily screened using gene expression microarray technique.
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BACKGROUND:β-catenin is the most critical signaling molecule in the Wnt/β-catenin signaling pathway, which is involved in the regulation of cellproliferation, differentiation and tissue self-healing balance. OBJECTIVE:To construct a stableβ-catenin over-expression lentivirus-mediated vector and to transfect mesenchymal stem cells line for investigating its effects on proliferation and migration of mesenchymal stem cells. METHODS:Over-expression vector, PLV-EF1A-catenin-RFP, was constructed and transfected the 293T cellto infect mesenchymal stem cells, and positive cells were selected with puromycin. The up-regulated efficiency of targetingβ-catenin gene at mRNA level was detected by real-time quantitative PCR, the effect on proliferation of mesenchmal stem cellwas assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve, and the migration ability was detected by Transwel motility assay. RESULTS AND CONCLUSION:The lentiviral vector targetingβ-catenin gene was constructed successful y, and a stable mesenchymal stem cellline that up-regulatedβ-catenin was established. Real-time quantitative PCR results showed that the expression ofβ-catenin gene was efficiently up-regulated by infecting PLV-EF1A-catenin-RFP (P<0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve showed that celldoubling time was shortened after infected with pLV-EF1A-catenin-RFP (P<0.05), indicating that the over-expression of theβ-catenin gene successful y increased the proliferative capability of mesenchymal stem cells. The Transwel assay also showed similar increasing results on the migration ability (P<0.01). The lenvivirus-mediated over-expression of theβ-catenin gene can be used to increase the proliferation and migration abilities of the mesenchymal stem cells.
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Objective To construct a lentiviral vector over-expressing β-catenin gene by multisite Gateway technology and confirm its effect.Methods By using multisite Gateway clone technique,the entry clone of pDown-Ctnnb1 was constructed using BP recombination reaction.Then,LR recombination reaction was performed among pUp-EF1A,pDown-Ctnnb1,pTail-IRES/DsRed-Express2 and pLV.Des3d.P/puro to generate an expression clone of pLV.EX3d.P/puro-EF1A＞Ctnnb1 ＞IRES/DsRed-Express2.In each step,PCR and sequencing analysis were used to verify the constructions.When it was verified that plasmids were transfected into 293T cells,PT-PCR was performed to determine the mRNA level of β-catenin gene.Results Both PCR and sequencing analysis revealed that β-catenin over-expression gene was inserted into the target site and the insertion sequence was perfectly corrected.The RT-PCR results showed that the expression of β-catenin gene was significantly upregulated.Conclusion The lenvivirus-mediate β-catenin over-expression gene was successfully constructed..
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Objective To investigate the expression and significance of TLR4, TLR9 and DC-SIGN in primary and recurrent condyloma acuminatum (CA) lesions. Methods An immunohistochemical method using streptavidin-peroxidase (SP) was performed to detect the expressions and distribution of TLR4, TLR9 and DC-SIGN in tissue specimens obtained from the recurrent CA lesions of 30 patients, primary CA lesions of 30 patients, and from the foreskin of 20 normal human controls. Results The expression levels of TLR4, TLR9 and DC-SIGN in primary and recurrent CA lesions were significantly higher than those in normal control tissue (all P < 0.001), and the cells expressing TLR4, TLR9 or DC-SIGN were mainly located in the basal and spinous layer in CA lesions. There was no significant difference in the expressions of TLR4, TLR9 or DC-SIGN between primary and recurrent CA lesions (all P> 0.05). A positive correlation was found between the expression of TLR4, TLR9 and DC-SIGN in CA lesions. Conclusion The overexpression of TLR4, TLR9 and DC-SIGN probably plays an important role in the occurrence and recurrence of CA.
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Objective To investigate the effect of transplantation of allogeneic bone marrow hematopoietic cells(HCs)and mesenchymal stem cells(MSCs)on experimental colitis(EC)in rats.Methods The HCs and MSCs obtained from SD male rats were cultured and expanded in vitro.In experiment 1 and 2 groups,HCs were labeled with bromodeoxyuridine(BrdU)and MSCs were obtained using the tube wall attach technique,respectively.Seventy-two female rats were infused with trinitrobenzene sulfonic acid(TNBS)to induce EC models.After 24 hours,HC or MSC suspensions were injected into the rats in experimental 1(n=18)and 2(n=18)groups via caudal veins,respectively.Control animals were injected with isotonic saline.The whole colon was removed on day 7,14 and 21 after transplantation and examined histopathologically.BrdU labeled HCs were tested with immunohistochemical staining and MSCs were detected for sex-determining gene(sry)by PCR.Results EC models were successfully established.The HCs or MSCs grew rapidly in the culture suspension.On day 7,14 and 21 after transplantation,the BrdU immunoreactive cells were detected in the colon(6/6),and the positive expression of the sry gene was found in 1/6,2/6 and 3/6,respectively.No positive labeled cell was found in controls.There was no significant improvement in histopathological scores on the colon in two experimental groups compared with the controls.Conclusions Allogeneic HCs and MSCs may localize in the colon of EC models.The ability of localization is higher in HCs than MSCs.The transplantation of HCs and MSCs can not obviously improve histopathologically.
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Objective To explore the dynamic changes of CD34 + cells and T lymphocyte subsets and best time of harvesting peripheral blood stem cell when G-CSF was used for peripheral blood stem cell mobilization in donors and patients. Methods A total of 12 donors and 16 patients who received chemotherapy for 7 d were injected G-CSF of 300 ?g/d to mobilize peripheral blood stem cells for 5 d and flow cytometry were used to detect the changes of CD34 + cells and T lymphocyte subsets everyday for 5 d. Results ① Before G-CSF treatment, there were obvious differences in bone marrow CD34 + cells between patients and donors (P
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AIM: The effects of selenium dioxide (SeO_2) on proliferation, apoptosis, intracellular reactive oxygen species (ROS) and Ca~(2+) levels in three leukemia cell lines NB4, K562 and HL-60 were investigated. METHODS: Three leukemia cell lines were treated with 3-30 ?mol/L SeO_2. Flow cytometry was used to detect apoptosis rate, and analyze the changes of ROS and Ca~(2+) level within cells. RESULTS: SeO_2 at 10 and 30 ?mol/L inhibited proliferation in three leukemia cell lines. Treatment with 30 ?mol/L SeO_2 for 48 h induced 54.0%, 46.5%, 49.6% apoptosis in NB4, K562, and HL-60 cells, respectively, and also markedly decreased ROS and Ca~(2+) levels among three cell lines. The rate of ROS positive cells in NB4 and HL-60 decreased with the increase in SeO_2 concentrations. ROS was clearly reduced with 30 ?mol/L SeO_2 in K562. Ca~(2+) levels were tardily declined with 10, 30 ?mol/L SeO_2 in NB4 and HL-60 cells. Ca~(2+) levels were clearly reduced with 30 ?mol/L SeO_2 in K562. CONCLUSION: SeO_2 induces apoptosis in three leukemia cells. The declines of intracellular ROS and Ca~(2+) levels are involved in apoptosis induced by SeO_2.