The antitumor effect of sodium morrhuate (SM ) and cis - diaminoaichloroplatinum (CDDP ) for gastric cancer cell SGC - 7901 in vitro is observed with MTT assay. Human gastric cancer cells line SGC - 7901 were inoculated subcutaneously in the naked mice , so xenografts were established. The tumor of the mice in the combination groups were cross injected with SM 50mg . kg~(-1) . time~(-1) , 50?l and CDDP ( 3mg ? kg~(-1)? time~(-), 50?l)in varying order at an interval of 30 minutes. In each mouse the tumors were injected every third davs for a total of three injections. lt was found SM had strong antitumor effect on gastric can- cer cells in vitro experiment. The antitumor effect of local injection in combination groups were much stronger than that in any other single drug groups. The histopathological section observation showed that all tumors cells were necrotic in combination groups, There could be seen fibrous histocysts around the . necrotic tumor tissue in observation groups , and necrotic vascular endotheliocytes. The experiment showed that combined injection with SM and CDDP had a synergic antitumor effect.

Objective:To observe the clinical efficacy of acupuncture combining with temperature sensitive point moxibustion therapy on laryngeal cough with yin-deficiency of lung-kidney. Methods:Divided 80 patients into test group and control group. Each included 40 people. The test group adopted the therapy by acupuncture kaiyin 1,Tianrong point,combining with temperature sensitive point (this point ofen appears in Yongquan Point Branch) moxibustion therapy,and the control group took the Western medicine conventional treatment. Then calculated the total effective rates of the two groups and analyzed statistically. Results:At the end of treatment,the total effective rate of test group was 95%,and the control group's was 85%,the difference between total effective rate was signifi cant difference (P

Objective: To study the role of vascular endothelial growth factor (VEGF) during angiogenesis in oral squamous cell carcinoma (SCC).Methods:Surgical specimens from 45 patients with oral SCC were examined for VEGF expression and microvessel density(MVD) by immunocytochemical staining. Anti-VEGF polyclonal antibody was used to determine VEGF expression and anti-CD 34 monoclonal antibody was used to determine MVD. Results:①VEGF protein was mainly found in cytoplasm of cancer cells and in few endothelial cells. Stained microvessels were mainly distributed in the regions of the connective tissue near the cancer nest;②there was a close positive correlation between MVD and VEGF expression;③SCC with high MVD or high VEGF level was mostly found with metastasis in cervical lymph node (P

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.

To investigate the dynamic homing process and characteristics of macrophages in different organs of immune-mediated aplastic anemia (AA) model mice. Macrophages in donor lymph nodes were sorted by magnetic beads and labeled with PKH67. After modeling according to the preparation method of the AA model, peripheral blood rountine analysis, bone marrow biopsy and HE staining results were analyzed to verify the modeling effect. On days 4, 8, and 12 of modeling, the bone marrow, spleen, and lymph node mononuclear cells were collected, and dynamic changes of PKH67-labeled macrophages in donor mice were analyzed by flow cytometry. In this study, dynamic changes in PKH67-labeled macrophages in the pathogenesis of AA model mice were explored. Macrophages in donor mice homed to the lymph nodes, expanding and differentiating in the lymph nodes, and finally transported to the bone marrow and spleen. Through proteomics mass spectrometry analysis, the related immune inflammatory response pathway of macrophages involved in the activation of the AA bone marrow microenvironment was preliminarily revealed, which provides a basis for the pathological macrophages involved in the pathogenesis of AA model mice.

Objective:To evaluate the efficacy of ultrasound-guided bilateral erector spinae plane (ESP) block for pediatric patients undergoing Nuss procedure with general anesthesia.Methods:Thirty-two American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients, aged 4-15 yr, undergoing Nuss procedure with general anesthesia, were divided into ESP block plus general anesthesia group (group EG, n=16) and general anesthesia group (group G, n=16) using a random number table method.Bilateral ESP blocks were performed after anesthesia induction in group EG.Intravenous analgesia was performed postoperatively in both groups.Face Legs Activity Cry Consolability (FLACC) scores both at rest and during coughing were recorded on admission to postanesthesia care unit (PACU) and at 1, 6, 12, 24 and 48 h after operation.When FLACC score at rest >4, ibuprofen 5.0-7.5 mg/kg was taken orally according to the requirements of pediatric patients and their parents.The intraoperative amount of propofol and remifentanil used and postoperative requirement for fentanyl and ibuprofen were recorded.The development of postoperative hypoxia (SpO 2 <90%), nausea and vomiting, and urinary retention was recorded.The development of puncture site infection, hematoma and subcutaneous emphysema was recorded.The tracheal extubation time, duration in PACU and length of postoperative hospital stay were also recorded. Results:Compared with group G, FLACC scores at rest within 12 h after operation and during coughing within 6 h after operation were significantly decreased, the intraoperative consumption of remifentanil was reduced, the postoperative requirement for fentanyl and ibuprofen was decreased, the incidence of hypoxemia was decreased, the tracheal extubation time and duration in PACU were shortened ( P<0.05), and no significant change was found in the consumption of propofol, incidence of nausea and vomiting, or length of postoperative hospital stay in group EG ( P>0.05). Conclusion:Ultrasound-guided bilateral ESP block can reduce the consumption of opioids in the perioperative period and enhance the analgesic efficacy with a higher safety, which is helpful in promoting short-term outcomes for pediatric patients undergoing Nuss procedure with general anesthesia.