Objective To investigate the effects of continuous venovenous hemodiafiltration (CVVHDF) on plasma lipopolysaccharide (LPS),interleukin(IL) 1β,IL-4,high mobility group box 1 (HMGB-1) in multiple organ dysfunction syndrome (MODS) dogs,to explore the therapeutic mechnism of CVVHDF.Methods Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to establish MODS model,then they were randomly divided into 2 groups:CVVHDF group (n=6) and MODS group (n=6).After endotoxin injection completed,the CVVHDF group received CVVHDF treatment for 24 h; MODS group did not receive CVVHDF treatment.The LPS,IL-1β,IL-4,HMGB-1 levels in plasma and ultrafiltrate were measured at different time points(before operation,before intravenous infusion of endotoxin,0 h,3 h,6 h,9 h,12 h,18 h,24 h after intravenous infusion completion of endotoxin),then the sieving coefficient (SC) was calculated.Results Compared with MODS group,the LPS,IL-1β,IL-4,HMGB-1 levels in plasma were significantly decreased in the CVVHDF group at 9 h,12 h,18 h,24 h (P ＜ 0.05).Some concentration of IL-1β,IL-4 and HMGB-1 was detected in the ultrafiltrate,the SC for IL-1β,IL-4 and HMGB-1 was 0.60±0.12,0.83±0.11,0.70±0.09 respectively,and LPS could not be detected in the ultrafiltrate,its SC was 0.After treatment,the damage level of lung and renal had been significantly reduced in CVVHDF group.Conclusion CVVHDF can effectively reduce the levels of pro-inflammatory mediator and anti-inflammatory cytokine related to MODS,cut off the vicious circle of cytokine network and reduce the damage level of organism in the treatment of MODS.

Objective:To investigate the adjusting role of Chinese herb purpura No.1 on the immune function of henoch-schonlein purpura(HSP).Methods:100 HSP children were divided into Chinese herb group and western medicine group with 1∶1,observing the effects of two groups and compare T lymphocyte subgroups such as CD3,CD4,CD8,CD4/CD8 and the level of IgG,IgA,IgM of two groups between before-treatment and after-treatment.Results:The effect and the adjusting role on the HSP children′s immune function of Chinese herb group was better than that of western medicine group while the recurrence rate of Chinese herb group was lower than that of western medicine group.Conclusion:Chinese herb can adjust the immune function and condense the symptoms′ duration time of HSP children. It also can decrease the recurrence rate of HSP children.

0 05); The protein expression of erbB2 in carcinomas of palate and floor of mouth were obviously higher than the expression in lip cancer and gingival carcinoma( P 0 05).Conclusion The amplification and overexpression of c erbB2 and p53 play important roles in the formation and development of OSCC, the overexpression of c erbB2 protein has more important significance in tumorigenesis of carcinomas of palate and floor of mouth; Not only transcriptional regulation but also translational modulation exists on the expression of c erbB2 and p53. [

Objective Plasma phospholipid metabolism analysis was employed to investigate plasma metabolic profile of acute renal graft rejection in order to find potential disease biomarkers and reveal its pathophysiological changes.Methods Selected 33 patients which did renal transplant in our hospital during 2009 to 2010,named preoperative group.14 patients were diagnosed as acute graft rejection,and 19 patients were non-acute graft rejection.Then use ultra performance liquid chromatography (UPLC) coupled with quadrupole time of flight mass spectrometry (qTOF-MS) to establish the metabolic fingerprint in those patients.The date was preprocessed with software Markerlynx,and analyzed with partial least squares discriminant analysis (PLS-DA),and data from conventional laboratory techniques were used for assessing renal function.Results In the plasma of renal transplant patients,seventeen biomarkers were discovered:creatinine,four sphingomyelins (SM),nine phosphatidylcholines (PC),three lysophosphatidylcholine (LPC).While the creatinine was increased,the others were reduced obviously.Conclutions UPLC-qTOF-MS based metabonomics can analyze the plasma phospholipid metabolism in patients with renal transplantation,reveal the law of phospholipids metabolic changes in the process of acute rejection.The levels of plasma phosphatidylcholine's esterlysis and fatty acids' β-oxidation are increased.The immunosuppressive therapy can reduce the activity of phospholipase,inhibit the level of plasma phosphatidylcholine's esterlysis and fatty acids' β-oxidation,which restraines the development of acute rejection after renal transplantation.Therefore it may be a promising new technology in diagnosing acute renal graft rejection after renal transplantation.

Objective To observe the members' dynamic change of renin-angiotensin system (RAS) and angiotensin converting enzyme 2(ACE2)-Ang(1-7)-Mas axis during the continuous venovenous hemodiafiltration (CVVHDF) treating the dogs with multiple organ dysfunction syndrome (MODS),and to investigate the efficacy mechanism on cardiac function.Methods Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to establish MODS model,then they were randomly divided into 2 groups:CVVHDF group (n=8) and MODS group (n=6).After endotoxin injection completion,the CVVHDF group received CVVHDF for 12 h,MODS group didn't.Radioimmunoassay,euzymelinked mmunosorbent assay (ELISA) were used to detect the content of renin,Ang Ⅰ,Ang Ⅱ,Ang (1-7).Real-time PCR was used to detect the expression of renin mRNA and ACE2 mRNA.Western blotting was used to detect the protein content of renin,Ang Ⅱ,ACE2 and Ang (1-7).Results (1) Organ function:Compared with the MODS group,the vital signs,heart,lung and renal function were significantly ameliorated in the CVVHDF group,the difference had statistical significance (P＜ 0.05).(2) RAS changes:To detect index from the level of organs,genetic and molecular in arterial tissue,the results displayed that the content of renin,Ang Ⅰ,Ang Ⅱ,the expression of renin mRNA,the protein content of renin,Ang Ⅱ in CVVHDF group were lower than that in MODS group,the difference had statistical significance (P ＜ 0.01).(3)ACE2-Ang(1-7)-Mas axis's changes:Using the same methods above to detect corresponding indicators,the results displayed that the content of ACE2,Ang(1-7),the expression of ACE2 mRNA and the protein content of ACE2,Ang(1-7) in CVVHDF group were significantly improved than that in MODS group,the difference had statistical significance (P ＜ 0.01).Conclusions In the process of CVVHDF treating MODS,the ACE2 -Ang(1-7)-Mas axis plays the role which antagonizes the RAS system,and to protect the cardiac function.This research may be important for MODS' clinical rescue.

Objective:To investigate the preoperative diagnostic value of 99Tc m-methoxyisobutylisonitrile (MIBI) planar imaging and SPECT/CT imaging for primary hyperparathyroidism (PHPT), and analyze the relevant factors affecting the imaging results. Methods:From June 2016 to September 2019, a total of 62 patients (15 males, 47 females, age range: 27-80 years) confirmed as PHPT by postsurgical pathology in Affiliated Hospital of Qingdao University were retrospectively enrolled. The diagnostic efficacies of 99Tc m-MIBI planar imaging and SPECT/CT imaging were compared using χ2 test. The differences of preoperative serum parathyroid hormone (PTH), Ca and the maximum diameter of lesion between the positive and negative groups of planar imaging were analyzed using independent-sample t test and Mann-Whitney U test. The region of interest (ROI) method was applied to calculate the uptake ratio of lesions to normal tissues at the early phase (T/Ne) and delayed phase (T/Nd) in positive cases of planar imaging. Pearson or Spearman correlation analysis was used to evaluate the correlation of T/Ne, T/Nd with preoperative serum PTH, Ca and the maximum diameter of lesion. The receiver operating characteristic (ROC) curves of preoperative serum PTH, Ca and positive planar imaging were drawn and the cut-off values were obtained. Results:The sensitivity of planar imaging and SPECT/CT imaging was 69.35%(43/62) and 87.10%(54/62) respectively ( χ2=5.729, P=0.017). The preoperative serum PTH, Ca levels and the maximum diameter of lesion in patients with positive planar imaging (253.32(107.00, 331.70) ng/L, 2.78(2.51, 2.87) mmol/L, (2.01±0.88) mm) were higher than those with negative planar imaging ((111.86±44.29) ng/L, (2.59±0.21) mmol/L, (1.42±0.55) mm; z values: -2.802, -1.978, t=3.300, all P<0.05). T/Ne was positively correlated with preoperative serum PTH ( rs=0.511, P<0.001) and the maximum diameter of lesion ( r=0.381, P=0.012), and T/Nd was positively correlated with preoperative serum PTH ( rs=0.538, P<0.001), Ca ( rs=0.348, P=0.022) and the maximum diameter of lesion ( r=0.463, P=0.002). The area under the ROC curve between preoperative serum PTH, Ca and planar imaging was 0.725 and 0.646, respectively. Preoperative serum PTH had a better predictive value with the optimal cut-off value of 150.4 ng/L. Conclusions:Preoperative serum PTH, Ca and the maximum diameter of lesion are positively correlated with 99Tc m-MIBI uptake in PHPT patients with positive planar imaging results. When preoperative serum PTH is lower than 150.4 ng/L, planar imaging is prone to false negative. SPECT/CT imaging has a significant value in preoperative diagnosis and the combination of PTH and CT can improve the positive rate.

Objective: To explore the effects and possible mechanism of rapamycin (RAPA) on apoptosis of CD4⁺CD25⁺ Tregs from the mouse severe aplastic anemia (SAA) model. Methods: The BALB/c female SAA model mice were induced by interferon-gamma in combination with busulphan. The SAA model mice were intraperitoneal injection with RAPA at daily dose of 0.5 mg/kg for 5 days (the RAPA-treated group, n=15) in the SAA group (n=15) and the un-treated group (n=15) were control. Bone marrow hematopoiesis changes were observed by the patho-morphological examination of femurs. The mononuclear cells of the peripheral blood and spleen were subjected to assess the intracellular Foxp3 expression in CD4⁺CD25⁺ Tregs by flow cytometry (FCM). In addition, after being pured by immunomagnetic beads, the splenic CD4⁺CD25⁺ Tregs was subjected to assess apoptosis by FCM and the Akt and Stat3 phosphorylation by using of western blot. Results: The patho-morphological examination of femurs showed normal marrow cell proliferation in un-treated group and hypocellularity in both SAA group and RAPA-treat group, with an increase in the number of fat cells. The bone marrow hematopoietic tissue ratio in RAPA-treat group was higher than SAA group [(9.75±1.83)% vs (7.00±2.00)%, Δx=2.15% (95%CI 0.15%-5.35%), P=0.037]. In the SAA group, FCM analysis showed down-expression of Foxp3 in CD4⁺CD25⁺ Tregs compared with the un-treated group. However, after treatment with RAPA, the expression of Foxp3 in CD4⁺CD25⁺ Tregs was increased (P<0.017). Compared with the un-treated group, increased CD4⁺CD25⁺ Tregs apoptosis [(19.84±1.39)% vs (29.85±2.72)%] with increased Akt phosphorylation accompanied by increased Stat3 phosphorylation was found in SAA group (P<0.05, respectively). On the contrary, RAPA-treated group exhibited CD4⁺ CD25⁺ Tregs with a reduction in apoptosis rate [(22.39±3.71)%], Akt phosphorylation and Stat3 phosphorylation compared with the SAA group (P<0.05, respectively). Conclusion These results indicate that RAPA may increase the expression of Foxp3 by down-regulation the levels of Akt and Stat3 phosphorylation and reduce apoptosis in splenic CD4⁺CD25⁺ Tregs from the mice model of SAA.

Objective: To investigate the response to ¹³¹I therapy and to explore the influence factors in postoperative differentiated thyroid carcinoma (DTC) patients with negative preablative stimulated thyroglobulin (psTg) and iodine-positive lymph node after the first radioablation. Methods: From May 2016 to October 2018, 108 DTC patients (28 males, 80 females, age: (45.7±10.4) years) with negative psTg who underwent ¹³¹I treatment for the first time in the Affiliated Hospital of Qingdao University were retrospectively enrolled. All patients had iodine-positive lymph nodes, which were showed by SPECT/CT imaging 5-6 d after ¹³¹I treatment. The treatment response was evaluated at 6-24 month after ¹³¹I treatment. Patients were divided into excellent response (ER) group and non-excellent response (non-ER) group according to the response to the first ¹³¹I treatment. Independent-sample t test, χ² test and Mann-Whitney U test were used to analyze differences of factors (e.g. age, gender, extraglandular infiltration) between the 2 groups, and then multivariate logistic regression was performed to find the influence factors for treatment response. The receiver operating characteristic (ROC) curve was employed to evaluate the predictive value of the iodine-positive lymph node size in ¹³¹I treatment response. Results: A total of 82 patients (75.93%, 82/108) achieved ER, and 26 (24.07%, 26/108) was non-ER patients. There were significant differences in age (t=-2.540, P=0.016), extraglandular infiltration (χ²=5.764, P=0.016), T stage (χ²=19.857, P<0.001), N stage (χ²=14.145, P=0.001), risk stratification of recurrence (χ²=11.487, P=0.003), ultrasound results before ¹³¹I treatment (χ²=44.819, P<0.001), dose of the first ¹³¹I treatment (U=780.0, P=0.018), size (long diameter) of iodine-positive lymph node (U=184.0, P<0.001), and psTg level (U=776.0, P=0.037) between ER and non-ER groups. Multivariate logistic regression showed that age, size of iodine-positive lymph node and ultrasound results before ¹³¹I treatment were closely related to ¹³¹I treatment response. The odds ratio (OR) values (95% CI) were 1.123 (1.025-1.231), 4.275 (1.893-9.653) and 260.86 (8.123-8 376.764), respectively. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value for non-ER were 100%(26/26), 70.73%(58/82), 77.78%(84/108), 52.00%(26/50) and 100%(58/58) respectively when the cut-off value of iodine-positive lymph node size was 5.5 mm. Conclusion The response of some DTC patients with negative psTg and iodine-positive lymph node after the first radioablation were non-ER. Age, ultrasound results before ¹³¹I treatment and size of iodine-positive lymph node are sensitive indicators for predicting clinical outcome in DTC patients with negative psTg and iodine-positive lymph node after the first radioablation.

Objective:To investigate the role of macrophages (M?) in the pathogenesis of modified immune-mediated aplastic anemia (AA) mice model.Methods:Before the establishment of the F1 AA mice model by total-body irradiation combined with allogeneic lymphocyte infusion, the mice of the CLO+AA group were treated with clodronate (CLO) liposomes to remove macrophages, and those of the PBS+AA group were treated with phosphate-buffered saline (PBS) liposomes and used as control. The severity of AA was observed by bone marrow (BM) pathological examination and peripheral blood cell count. Flow cytometry (FCM) was used to detect the CD4 +/CD8 + T lymphocyte subsets in the BM and M? subsets in the BM and spleen of each group. The levels of IFN-γ, TNF-α, G-CSF, GM-CSF, EPO, and TPO in the peripheral blood were detected using enzyme-linked immunosorbent assay. Finally, the relationships between inflammatory factors and M? subsets were analyzed. Results:The BM fatty conversion of mice in the CLO+AA group was significantly alleviated compared with the PBS+AA group. Hemoglobin counts were (91.50±31.63) and (110.65±24.15) g/L, respectively, and the platelet counts were (90.85±121.90) × 10 6/L and (461.13±483.45) ×10 6/L, respectively. The differences were all statistically significant (all P<0.05) . After removing macrophages, the proportions of CD4 + and CD8 + T lymphocytes in BM of mice in the CLO+AA group decreased, but the reduction of CD8 + T cells was more significant. The proportions of CD4 + T cells and CD8 + T cells in BM of the PBS+AA group were (18.5±10.17) % and (36.23±6.40) %, respectively, and in the CLO+AA group were (7.58±8.00) % and (6.67±5.78) %, respectively. Similarly, the percentage of macrophages in the spleen and BM in the CLO+AA group was significantly reduced compared with the PBS+AA group, most of which were M1 macrophages ( P<0.05) . The levels of IFN-γ in peripheral blood of the PBS+AA and CLO+AA groups were (602.37±104.62) ng/L and (303.01±87.22) ng/L, respectively, the levels of TNF-α were (34.46±1.42) ng/L and (23.25±4.21) ng/L, respectively, the levels of GM-CSF were (9.32 ± 2.00) ng/L and (64.85±12.25) ng/L, respectively, the levels of G-CSF were (5 891.78±2 632.39) ng/L and (17 784.16±488.36) ng/L, respectively, the levels of EPO were (9 667.31±4 501.95) ng/L and (2 078.02±897.56) ng/L, respectively, and the levels of TPO were (6.36±2.09) ng/L and (11.67±2.86) ng/L, respectively (all P<0.05) . Conclusions:This study confirmed that macrophages were involved in the pathogenesis of AA, and the degree of BM damage in AA mice was improved by removing macrophages in advance. The imbalance of M1/M2 macrophages and the changes of IFN-γ and TNF-α may be important mechanisms that eventually lead to AA.

Objective:This study aims, in addition to characterizing pathogenic T cells trafficking to bone marrow (BM) and other organs, to establish immune-mediated AA C.B10 mouse model by DsRed mouse (B6 background) lymph nodes (LN) cells infusion after a total body irradiation (TBI) .Methods:The C.B10 mice received a 5 Gy TBI and then were infused with DsRed mouse (B6 background) LN cells at 5×10 6/mouse via a tail vein injection. The severity of bone marrow failure (BMF) was observed by mononuclear cell count in bone marrow and peripheral blood cell count. On days 3, 6, 9, and 12, mice were sacrificed and collected BM, spleens, LN, or thymus to analyze the dynamic change and activation status of donor T cells in these organs by a flow cytometry. At day 12, the donor-derived T cells from BM, spleens, and LN were sorted to collect the DsRed +CD3 +CD4 + T cells and DsRed +CD3 +CD8 + T cells for RNA isolation and gene expression analyses by PCR array. Results:Relative to control animals that received 5 Gy TBI without LN cell infusion, AA mice developed severe BMF with dramatic decrease in total BM cells, hemoglobin, white blood cells, and platelets in peripheral blood on days 9 and 12 after the LN cell infusion. The frequencies of DsRed + T cells trafficking to BM, LN, and spleens increased with time. Surprisingly, although the DsRed + T cells in BM increased dramatically at a level much higher than those in the spleens and LN on day 12, there were very few DsRed + T cells in BM on days three and six, which was significantly lower than those in spleens or LN. The frequency of DsRed + T cells in thymus was the lowest during the whole process. On day 12, the DsRed +CD3 +CD4 + T cells of BM, LN, and spleens from AA mice were (91.38±2.10) %, (39.78±6.98) %, and (67.87±12.77) %, respectively. On the contrary, the DsRed +CD3 +CD8 +T cells of BM, LN, and spleens were (98.21±1.49) %, (94.06±4.20) %, and (96.29±1.23) %, respectively. We assessed the donor T cell phenotypes using the CD44 and CD62L markers and found that almost all of the DsRed +CD4 + or DsRed +CD8 + T cells in BM were effector memory T cell phenotype from day 9 to day 12. Meanwhile, transcriptome analyses showed higher expression in CD38, IFN-γ, LAG3, CSF1, SPP1, and TNFSF13B in BM DsRed +CD4 + and DsRed +CD8 + T cells. However, there was a lower expression in FOXP3 and CTLA4 in BM DsRed +CD4 + T cells than those in spleens and LN. Conclusions:The DsRed LN cells infusion to induce BMF in CB10 mice enabled to track the donor-derived pathogenic T cells. Besides previously published findings in this model, we demonstrated that donor CD4 + and CD8 + T cells primarily homed to spleens and LN, expanded and differentiated, then infiltrated in BM with a terminal effector memory phenotype. The T cells infiltrated in BM showed more activation and less immunosuppression characteristics compared to those homing to spleens and LN during the BMF development.