[Single non-blood-related umbilical cord blood transplantation using a reduced-intensity conditioning regimen for the treatment of severe aplastic anemia].

Objective: To evaluated the clinical efficacy of a reduced-intensity preconditioning regimen for single non-blood-related umbilical cord blood transplantation (sUCBT) in the treatment of severe aplastic anemia (SAA) . Methods: The clinical data of 63 patients with SAA who underwent sUCBT from January 2021 to July 2023 at the Department of Hematology of the First Affiliated Hospital of USTC were retrospectively analyzed. Fifty-two patients received total body irradiation/total bone marrow irradiation (TMI) combined with fludarabine or a cyclophosphamide- conditioning regimen (non-rATG group) , while 11 patients received rabbit anti-human thymocyte immunoglobulin (rATG) combined with TMI, fludarabine, or the cyclophosphamide-conditioning regimen (rATG group) . All patients received cyclosporine A and mycophenolate mofetil for graft-versus-host disease (GVHD) prophylaxis. Complications post-transplantation and long-term survival were compared between the two groups. Results: The baseline parameters were balanced between the two groups (P>0.05) . In the rATG group, all patients achieved stem cell engraftment, and in the non-rATG group, five patients had primary graft failure. There was no significant difference in the cumulative incidence of neutrophil engraftment at 42 days after transplantation or platelet engraftment at 60 days between the two groups. The incidence of grade Ⅱ-Ⅳ acute GVHD in the rATG group was significantly lower than in the non-rATG group (10.0% vs. 46.2% , P=0.032) , and the differences in the cumulative incidences of grade Ⅲ/Ⅳ acute GVHD and 1-year chronic GVHD were not statistically significant (P=0.367 and P=0.053, respectively) . There were no significant differences in the incidences of pre-engraftment syndrome, bacterial bloodstream infections, cytomegalovirus viremia, or hemorrhagic cystitis between the two groups (P>0.05 for all) . The median follow-up time for surviving patients was 536 (61-993) days, and the 1-year transplantation related mortality (TRM) of all patients after transplantation was 13.0% (95% CI 6.7% -24.3% ) . Among the patients in the non-rATG and rATG groups, 15.5% (95% CI 8.1% -28.6% ) and 0% (P=0.189) , respectively, had mutations. The 1-year overall survival (OS) rate of all patients after transplantation was 87.0% (95% CI 75.7% -93.3% ) . The 1-year OS rates in the rATG group and non-rATG group after transplantation were 100% and 84.5% , respectively (95% CI 71.4% -91.9% ) (P=0.198) . Conclusion: The preliminary results of sUCBT with a low-dose irradiation-based reduced-intensity conditioning regimen with fludarabine/cyclophosphamide for the treatment of patients with SAA showed good efficacy. Early application of low-dose rATG can reduce the incidence of acute GVHD after transplantation without increasing the risk of implantation failure or infection.

[High expression of secretogranin II increases oxaliplatin resistance of colorectal cancer cells].

OBJECTIVE: To investigate the expression of secretogranin II (SCG2) in colorectal cancer (CRC) tissues and its impact on oxaliplatin resistance of CRC cells. METHODS: We performed immunohistochemistry to detect the expression level of SCG2 on a tissue microarray containing 96 CRC and 84 adjacent tissues and analyzed the association of SCG2 expression with the clinical features of the CRC patients. SCG2 expression was also measured in DLD1 cells treated with oxaliplatin using immunoblotting and RT-qPCR analyses. The effects of SCG2 expression on oxaliplatin sensitivity and cell viability were evaluated in a DLD1 cell model of SCG2 knockout established using CRISPR-cas9 technique, and the expressions of apoptosis-related proteins were detected using Western blotting and RT-qPCR. We further examined SCG2 expression levels in an oxaliplatin-resistant DLD1 cell line and its parental DLD1 cells. RESULTS: SCG2 expression was significantly increased in CRC tissues as compared with the adjacent tissues (1.932±0.816 vs 1), and the tumor tissues in advanced stages showed higher SCG2 expression levels. In DLD1 cells, treatment with oxaliplatin significantly increased SCG2 expression, and SCG2 knockout obviously increased oxaliplatin sensitivity of the cells and enhanced the expressions of apoptosis-related proteins. Compared with the parental cells, oxaliplatin-resistant DLD1 cells showed a significant increase of SCG2 expression by 3.901±0.471 folds. CONCLUSION: SCG2 may serve as a risk gene in CRC, and its high expression increases oxaliplatin resistance of CRC cells.

[Prognostic value of perioperative change of neutrophil elastase and myeloperoxidase in coronary circulation on perioperative myocardial injury and clinical outcome of patients underwent surgical valve replacement].

Objective: To investigate the clinical value of observing perioperative changes of myeloperoxidase (MPO) and neutrophil elastase (NE) in coronary artery circulation in patients underwent valve replacement surgery. Methods: This perspective cohort study was performed in patients who underwent valvular surgery in Nanjing Drum Tower Hospital and Fuwai Hospital from June 2021 to June 2022. Patients were divided into perioperative myocardial injury group and age-, sex- and type of cardiac procedure-matched non-perioperative myocardial injury control group in the ratio of 1∶1. Perioperative myocardial injury was defined as cardiac troponin T (cTnT)>0.8 µg/L on the first postoperative day (POD), and the cTnT level on the second POD increased by more than 10% compared with the cTnT level on the first POD. During the operation, blood samples were collected from the coronary sinus before clamping ascending aorta, and within 5 minutes after de-clamping ascending aorta. Then, the levels of MPO and NE on coronary sinus were continuously measured. The death, severe ventricular arrhythmia, pneumonia, re-intubation, repeat cardiac surgery, extracorporeal membrane oxygenation (ECMO), intra-aortic balloon pump (IABP), continuous renal replacement therapy (CRRT), mechanical ventilation time and the duration of intensive care unit (ICU) were recorded. The levels of MPO and NE and the incidence of clinical outcomes were compared between the myocardial injury group and the control group. The independent risk factors of myocardial injury were analyzed by multivariate logistic regression. Results: A total of 130 patients were enrolled, aged (60.6±7.6) years old, with 59 males (45.4%). There were 65 patients in the myocardial injury group and 65 patients in the control group. During hospitalization, there was no death, ECMO, IABP and CRRT cases in both groups. Compared with the control group, the incidence of severe ventricular arrhythmia (13.8%(9/65) vs. 3.1%(2/65), P=0.03), pneumonia (20.0%(13/65) vs. 3.1%(2/65), P=0.03), re-intubation (6.2%(4/65) vs. 0, P=0.04) was significantly higher in myocardial injury group. The mechanical ventilation time (16.8(10.7, 101.7) h vs. 7.5(4.7, 15.1) h, P<0.01), and the duration of ICU (3.7(2.7, 18.9) vs. 2.7(1.8, 6.9)d, P<0.01) were significantly longer in myocardial injury group compared with the control group. There was no significant difference in the levels of MPO and NE in coronary sinus blood between the two groups before aortic clamping (all P>0.05). However, MPO ((551.3±124.2) µg/L vs. (447.2±135.9) µg/L, P<0.01) and NE ((417.0±83.1)µg/L vs. (341.0±68.3)µg/L, P<0.01) after 5 min aortic de-clamping were significantly higher in myocardial injury group than in the control group. Multivariate logistic regression analysis showed that the levels of NE (OR=1.02, 95%CI: 1.01-1.02, P<0.01), MPO (OR=1.00, 95%CI: 1.00-1.01, P=0.02) and mechanical ventilation time (OR=1.03, 95%CI: 1.01-1.06, P=0.02) were independent risk factors of myocardial injury in patients after surgical valvular replacement. Conclusion: Perioperative myocardial injury is related poor clinical outcomes, perioperative NE and MPO in coronary artery circulation are independent risk factors of perioperative myocardial injury in patients undergoing valve replacement surgery.

Use of peripheral blood eosinophils to guide post-operative glucocorticoid therapy in patients with chronic rhinosinusitis with nasal polyps: a randomised, controlled trial.

OBJECTIVE: This study aimed to explore the utility of the eosinophil percentage in peripheral blood for guiding post-operative glucocorticoid therapy in patients with chronic rhinosinusitis with nasal polyps. METHODS: Forty-four patients with chronic rhinosinusitis with nasal polyps underwent functional endoscopic sinus surgery and were randomly divided into two groups. Patients in the standard treatment group used oral and nasal spray glucocorticoids. In the biomarker treatment group, patients with peripheral blood eosinophil percentage values less than 3.05 per cent did not receive glucocorticoid treatment, whereas patients with values 3.05 per cent or above were part of the standard treatment group. Visual Analogue Scale, Sino-Nasal Outcome Test-22 scores, endoscopic Lund-Kennedy scores, eosinophils, interleukin-5 and eosinophil cationic protein in peripheral blood, and nasal secretions were measured. RESULTS: After functional endoscopic sinus surgery, the Visual Analogue Scale, Sino-Nasal Outcome Test-22 and Lund-Kennedy scores were significantly reduced in both groups; there were no significant differences in those indicators between the groups during the three follow-up visits. CONCLUSION: Peripheral blood eosinophil percentage offers a potential biomarker to guide post-operative glucocorticoid therapy in patients with chronic rhinosinusitis with nasal polyps.

[Outcomes and prognosis of radical surgery in patients with stageâ b2 and â ¡a2 cervical squamous cell carcinoma].

Objective: To evaluate the survival, complications and prognostic factors in patients with stageⅠb2 and Ⅱa2 cervical squamous cell carcinoma treated by primarily radical surgery with or without postoperative adjuvant therapy. Methods: The clinical and pathological data of patients with stageⅠb2 and Ⅱa2 cervical squamous cell carcinoma treated in the Cancer Hospital of the University of Chinese Academy of Sciences from January 2015 to January 2018 were retrospectively analyzed. All patients underwent Querleu-Morrow classification (Q-M classification) C2 radical surgery, including extensive hysterectomy+pelvic lymphadenectomy with or without adjuvant therapy based on postoperative risk factors. Survival rate was calculated by Kaplan-Meier method and survival curve was drawn. Univariate analysis was performed by using the log-rank test to analyze the clinicopathological factors related to the prognosis of patients. Multivariate analysis was performed by using Cox regression method to analyze independent risk factors affecting survival prognosis. Results: (1) The median age of 643 patients with cervical squamous cell carcinoma was 50 years old (45-58 years old). Clinical stage: 260 cases (40.4%, 260/643) of stage Ⅰb2, 383 cases (59.6%, 383/643) of stage Ⅱa2. (2) Among 643 cases underwent Q-M classification C2 surgery, 574 cases (89.3%, 574/643) of them received adjuvant therapy and 184 cases (28.6%, 184/643) of them had grade 3-4 complications after treatment, including 134 cases (20.8%, 134/643) early complications and 66 cases (10.3%, 66/643) late complications. The incidence of grade 3-4 complications in 574 patients received postoperative adjuvant therapy was 30.1% (173/574), which was significantly different from that in 69 patients who received surgery alone (15.9%, 11/69; χ²=6.08, P=0.014). (3) All 643 cases were followed up, and the median follow-up time was 40 months (3-76 months). During the follow-up period, 117 cases (18.2%, 117/643) recurred, including 45 cases (7.0%, 45/643) of local recurrence, 54 cases (8.4%, 54/643) of distant metastasis, and 18 cases (2.8%, 18/643) of local recurrence and distant metastasis. The 5-year progression-free survival (PFS) and 5-year overall survival (OS) rates of patients with stage Ⅰb2 and Ⅱa2 cervical squamous cell carcinoma were 79.9% and 85.5%, respectively. Univariate analysis showed that pelvic lymph node metastasis, para-aortic lymph node metastasis, deep stromal infiltration, and lymph-vascular space invasion were significantly associated with 5-year PFS in patients with stage Ⅰb2 and Ⅱa2 cervical squamous cell carcinoma (all P<0.05). The maximum diameter of tumor, pelvic lymph node metastasis and para-aortic lymph node metastasis were significantly associated with the 5-year OS of cervical squamous cell carcinoma in stages Ⅰb2 and Ⅱa2 (all P<0.05). Multivariate analysis showed that pelvic lymph node metastasis and para-aortic lymph node metastasis were independent factors affecting 5-year PFS and 5-year OS in patients with stage Ⅰb2 and Ⅱa2 cervical squamous cell carcinoma (all P<0.01). Conclusion: Radical surgery is a feasible and effective primary treatment for stagesⅠb2 and Ⅱa2 cervical squamous cell carcinoma, with a high 5-year survival rate and an acceptable complication rate.

[Technical summary and experiences of urinary reconstruction in mouse kidney transplantation].

The present paper summarizes the methods and technical key points for urinary reconstruction in mouse kidney transplantation. Both bladder patch and ureter implantation techniques are feasible options for urinary reconstruction in mouse kidney transplantation. The dominant complication in bladder patch technique is ischemic necrosis of patch and distal ureter and is associated with donor mouse strains and surgical skills. The most common complication in ureter implantation technique is urine leakage and is related to recipient mouse gender and technical skills. The key technical points for bladder patch method are estimation of blood supply for bladder patch and properly trimming and suturing bladder walls. The key points for ureter implantation method are properly trimming and fixation of the ureter. Comparing with the bladder patch, the ureter implantation technique is faster, its complications are easier to be repaired by secondary surgery, but the risk of urine retention is higher at late time. The timeliness of surgical intervention is a key factor for successfully repairing the early urinary complications.

Osimertinib and anti-HER3 combination therapy engages immune dependent tumor toxicity via STING activation in trans.

Over the past decade, immunotherapy delivered novel treatments for many cancer types. However, lung cancer still leads cancer mortality, and non-small-cell lung carcinoma patients with mutant EGFR cannot benefit from checkpoint inhibitors due to toxicity, relying only on palliative chemotherapy and the third-generation tyrosine kinase inhibitor (TKI) osimertinib. This new drug extends lifespan by 9-months vs. second-generation TKIs, but unfortunately, cancers relapse due to resistance mechanisms and the lack of antitumor immune responses. Here we explored the combination of osimertinib with anti-HER3 monoclonal antibodies and observed that the immune system contributed to eliminate tumor cells in mice and co-culture experiments using bone marrow-derived macrophages and human PBMCs. Osimertinib led to apoptosis of tumors but simultaneously, it triggered inositol-requiring-enzyme (IRE1α)-dependent HER3 upregulation, increased macrophage infiltration, and activated cGAS in cancer cells to produce cGAMP (detected by a lentivirally transduced STING activity biosensor), transactivating STING in macrophages. We sought to target osimertinib-induced HER3 upregulation with monoclonal antibodies, which engaged Fc receptor-dependent tumor elimination by macrophages, and STING agonists enhanced macrophage-mediated tumor elimination further. Thus, by engaging a tumor non-autonomous mechanism involving cGAS-STING and innate immunity, the combination of osimertinib and anti-HER3 antibodies could improve the limited therapeutic and stratification options for advanced stage lung cancer patients with mutant EGFR.

[Correlation between immune reconstitution and chronic graft-versus-host disease after unrelated cord blood transplantation and sibling peripheral blood stem cell transplantation].

Objective: To explore the relationship between the reconstitution of immune cells in patients with hematological malignancies and the occurrence of chronic graft-versus-host disease (cGVHD) after treatment with unrelated cord blood transplantation (UCBT) and sibling peripheral blood stem cell transplantation (PBSCT) . Methods: A total of 124 patients undergoing allogenic hematopoietic stem cell transplantation (allo-HSCT) in the First Affiliated Hospital of University of Science and Technology of China from March 2018 to August 2019, including 96 patients with UCBT and 28 patients with PBSCT. Peripheral blood immune cells of patients with UCBT and PBSCT were detected at 1, 3, 6, 9, and 12 months after transplantation using flow cytometry, and both UCBT and PBSCT patients were divided into cGVHD and non-cGVHD groups based on whether cGVHD occurred to explore the correlation between the immune cells reconstitution of the two types of transplantation and cGVHD. Results: ①The cumulative incidence of the moderate to severe cGVHD in the UCBT group was significantly lower than that in the PBSCT group[9.38% (95% CI 3.35%-15.02%) vs 28.57% (95% CI 9.72%-43.50%) , P=0.008]; the 2-year cumulative incidence of cGVHD and moderate to severe cGVHD in the UCBT group was lower than that in the PBSCT group[15.60% (95% CI 9.20%-23.60%) vs 32.10% (95% CI 15.80%-49.70%) , P=0.047; 10.40% (95% CI 5.30%-17.50%) vs 28.60% (95% CI 13.30%-46.00%) , P=0.014]. ②The absolute counts of CD4(+)T cells in the UCBT group were higher than those in the PBSCT group at 6, 9, and 12 months after transplantation[59.00 (36.70-89.65) ×10(7)/L vs 31.40 (18.10-44.00) ×10(7)/L, P<0.001; 71.30 (49.60-101.45) ×10(7)/L vs 41.60 (25.82-56.27) ×10(7)/L, P<0.001; 83.00 (50.17-121.55) ×10(7)/L vs 44.85 (31.62-62.10) ×10(7)/L, P<0.001]; the proportions of CD4(+)T cells in the UCBT group were always higher than those in the PBSCT group (P<0.05) . The absolute counts and proportions of B cells in the PBSCT group were higher than those in the UCBT group at the first month after transplantation[0.70 (0.30-1.70) ×10(7)/L vs 0.10 (0-0.30) ×10(7)/L, P<0.001; 0.45% (0.30%-2.20%) vs 0.20% (0.10%-0.40%) , P=0.002]; the absolute counts and proportions of B cells in the UCBT group were higher than those in the PBSCT group at 9 and 12 months after transplantation[53.80 (28.00-103.20) ×10(7)/L vs 23.35 (5.07-35.00) ×10(7)/L, P<0.001; 21.45 (11.80-30.45) % vs 9.00% (3.08%-16.73%) , P<0.001. 66.70 (36.97-98.72) ×10(7)/L vs 20.85 (7.72-39.40) ×10(7)/L, P<0.001; 22.20% (14.93%-29.68%) vs 8.75% (5.80%-18.93%) , P<0.001]. The absolute counts and proportions of regulatory B (Breg) cells in the UCBT group were higher than those in the PBSCT group at 6, 9, and 12 months after transplantation[1.23 (0.38-3.52) ×10(7)/L vs 0.05 (0-0.84) ×10(7)/L, P<0.001; 5.35% (1.90%-12.20%) vs 1.45% (0-7.78%) , P=0.002. 2.25 (1.07-6.71) ×10(7)/L vs 0.12 (0-0.77) ×10(7)/L, P<0.001; 6.25% (2.00%-12.33%) vs 0.80% (0-5.25%) , P<0.001. 3.69 (0.83-8.66) ×10(7)/L vs 0.46 (0-0.93) ×10(7)/L, P<0.001; 6.15% (1.63%-11.75%) vs 1.40% (0.18%-5.85%) , P<0.001].The absolute counts and proportions of CD3(+)T cells, CD8(+)T cells, and Treg cells in the UCBT group were not significantly different from those in the PBSCT group. ③The absolute counts of B cells in the non-cGVHD group of UCBT patients were higher than those in the moderate to severe cGVHD group at 6 and 12 months after transplantation (P=0.038, P=0.043) ; the proportions of B cells in the non-cGVHD group were higher than those in the moderate to severe cGVHD group at 6 months after transplantation (P=0.049) . The absolute counts of Breg cells in the non-cGVHD group of patients with UCBT were higher than those in the moderate to severe cGVHD group at 6, 9, and 12 months after transplantation (P=0.006, P=0.028, P=0.050) ; the proportions of Breg cells in the non-cGVHD group were higher than those in the moderate to severe cGVHD group at 9 months after transplantation (P=0.038) . ④The absolute counts and proportions of B and Breg cells in the non-cGVHD group of patients with PBSCT were not statistically different than those in the moderate to severe cGVHD group. Conclusion: In the process of immune cell reconstitution, the Breg cells in the UCBT group were higher than those in the PBSCT group, and the Breg cells in the non-cGVHD group of the two types of transplantation were always higher than those in the moderate to severe cGVHD group, indicating that Breg cells can reduce the occurrence of cGVHD, revealing the possible reason for the lower incidence of cGVHD in the UCBT group.

[The hearing outcomes of modified canal wall down and canal wall up mastoidectomy and tympanoplasty].

Objective: To compare the effect of hearing improvement after canal wall down mastoidectomy and tympanoplasty(CWDM) and canal wall up mastoidectomy and tympanoplasty(CWUM). Methods: 88 patients who underwent CWDM or CWUM in 2014-2017 with complete follow-up data were retrospectively studied. There were 45 males and 43 females, aged 12-68 years old, and the average follow-up time was 12.3 months. Among them, 42 cases underwent CWDM and 46 cases underwent CWUM. A series of improvements were made in the clinical practice of CWDM: (1) to expand the indications properly, including limited lesions with sclerotic mastoid and narrow tympanic sinus; (2) to form disciform cavity strictly during operation in order to retain the appropriate height of facial nerve crest; (3) to avoid the cleaning of mastoid cavity in the post-operation follow up, thus the operative cavity was effectively constricted; and (4) to transfer the temporal muscle flap in pneumatic mastoid to reduce the cavity. The condition of dry ear and the size of cavity were observed after operation. The average pure tone threshold (PTA) before/after operation and threshold changes of each frequency were compared between the two groups. SPSS 19.0 software was used to analyze the data. Results: The mastoid cavity of CWDM was significantly narrowed after operation and the width of the external auditory canal was close to the CWUM group. The postoperative PTA of the CWDM and CWUM group was reduced by 11.4 dB and 10.4 dB respectively, with no significant difference (t=0.290, P=0.770). The average value of bone conduction threshold after operation was reduced by 1.8 dB and 1.9 dB respectively, with no significant difference (t=-0.076, P=0.940). The mean value of ABG after operation was shortened by 9.6 dB and 8.4 dB respectively, with no significant difference (t=-0.370, P=0.712). The threshold decrease of 1 000 Hz of CWUM was slightly better than that of CWDM, and the threshold decreases of the other frequencies of CWDM were slightly better than those of CWUM, however, the differences were not statistically significant (P>0.05). Conclusion: Through the adjustment of indications and a series of intraoperative and postoperative improvements, CWDM can achieve the same effect of postoperative hearing improvement as CWUM.

IL-6 stimulates lncRNA ZEB2-AS1 to aggravate the progression of non-small cell lung cancer through activating STAT1.

OBJECTIVE: To illustrate the role of interleukin 6 (IL-6) in the progression of non-small cell lung cancer (NSCLC) via activating STAT1. PATIENTS AND METHODS: The level of IL-6 mRNA in 48 paired NSCLC tissues and matched normal ones was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Kaplan-Meier curves were depicted for assessing the overall survival of NSCLC patients with high or low level of IL-6 mRNA. Subsequently, the ZEB2-AS1 level in A549 cells treated with different doses of IL-6 for different time points was determined. After A549 cells were treated with different doses of IL-6, wound closure assays were performed to assess the migration of cells. Protein levels of pSTAT1 and STAT1 in IL-6-treated A549 cells were detected by Western blot. The regulatory effect of STAT1 on IL-6-induced migration of A549 cells was also evaluated. The interaction between ZEB2-AS1 and STAT1 was explored through Chromatin Immunoprecipitation (ChIP) assay. Finally, the role of ZEB2-AS1/STAT1 axis in regulating NSCLC cells was investigated through rescue experiments. RESULTS: Our results indicated that IL-6 was upregulated in NSCLC tissues and cancer cell lines. NSCLC patients with T3-T4 or accompanied with lymphatic metastasis had a higher IL-6 abundance than those with T1-T2 or without metastatic foci. The worse prognosis was identified in NSCLC patients with high expression of IL-6 compared to those with low expression. ZEB2-AS1 showed dose-dependent and time-dependent increase in IL-6-treated A549 cells. IL-6 treatment gradually enhanced the migration ability of A549 cells in a dose-dependent manner. In IL-6-treated A549 cells, protein level of pSTAT1 was remarkably upregulated, and knockdown of STAT1 significantly reversed the promotive effect of IL-6 on migration ability of A549 cells. The results of ChIP assay verified the interaction between ZEB2-AS1 and STAT1. In addition, ZEB2-AS1 could reverse the regulatory effect of STAT1 on the migration ability of A549 cells. CONCLUSIONS: IL-6 was upregulated in NSCLC and accelerated the progression of NSCLC via activating STAT1/ ZEB2-AS1.

[The experiment study of endovascular denervation in treatment of cancer pain].

Objective: To assess the feasibility and safety of endovascular denervation (EDN) with a multi-electrode radiofrequency ablation catheter on beagles. Methods: A total of 18 beagles, (10.2±1.1) kg,of either gender,were provided by the Animal Center of Southeast University (SYXK (Su) 2016-0013). They were divided equally into three groups:the instant euthanizing group, the long-term follow-up group and the sham operation group. Beagles in the instant euthanizing group were euthanized immediately after EDN. Beagles in the long-term follow-up group were euthanized three months after EDN. Beagles in the sham operation group underwent sham operation and were euthanized three months later. Blood biochemistry was measured at baseline, and immediately, 15 days, 30 days and 90 days after the surgery. Computerized tomographic (CT) angiography was determined before the surgery and 60 days after the surgery. Digital subtraction angiography (DSA) was determined 90 days after the surgery. Histopathologic analyses were used to identify the changes of arterial wall and neuron cells. Results: Beagles in the long-term follow-up group and the sham operation group all underwent EDN successfully without accidental death. No abdominal aortic perforation and peripheral tissue necrosis were found at Necropsy. No vascular injuries were found by CTA and DSA in each group. There was no statistical difference in hematological analyses, 90 days after the surgery:white blood cell:(12.5±1.5)×10(9)/L vs (13.2±0.7)×10(9)/L, P=0.275; red blood cell:(7.0±0.6)×10(9)/L vs (6.3±0.4)×10(9)/L, P=0.089; total bilirubin:(2.9±0.4) µmol/L vs (3.0±0.6) µmol/L, P=0.681; glutamic-pyruvic transaminase:(40±11) U/L vs (37±6) U/L, P=0.168; glutamic oxalocetie transaminase:(51±11) U/L vs (48±9) U/L, P=0.221; urea nitrogen:(7.2±1.2) mmol/L vs (6.9±0.8) mmol/L, P=0.505; creatinine:(60±9) µmol/L vs (59±9) µmol/L, P=0.81; prothrombin time:(7.2±0.7) s vs (7.0±0.7) s, P=0.719. Histopathological analyses showed that there were hypercellular appearance of nerve bundle and thickened perineurium in EDN groups, while normal perineurium around nerve bundle in the sham operation group. Conclusion: EDN could be applied in beagles safely and feasibly.

Biochemical bases of growth variation during development: a study of protein turnover in pedigreed families of bivalve larvae (Crassostrea gigas).

Animal size is a highly variable trait regulated by complex interactions between biological and environmental processes. Despite the importance of understanding the mechanistic bases of growth, predicting size variation in early stages of development remains challenging. Pedigreed lines of the Pacific oyster (Crassostrea gigas) were crossed to produce contrasting growth phenotypes to analyze the metabolic bases of growth variation in larval stages. Under controlled environmental conditions, substantial growth variation of up to 430% in shell length occurred among 12 larval families. Protein was the major biochemical constituent in larvae, with an average protein-to-lipid content ratio of 2.8. On average, 86% of protein synthesized was turned over (i.e. only 14% retained as protein accreted), with a regulatory shift in depositional efficiency resulting in increased protein accretion during later larval growth. Variation in protein depositional efficiency among families did not explain the range in larval growth rates. Instead, changes in protein synthesis rates predicted 72% of growth variation. High rates of protein synthesis to support faster growth, in turn, necessitated greater allocation of the total ATP pool to protein synthesis. An ATP allocation model is presented for larvae of C. gigas that includes the major components (82%) of energy demand: protein synthesis (45%), ion pump activity (20%), shell formation (14%) and protein degradation (3%). The metabolic trade-offs between faster growth and the need for higher ATP allocation to protein synthesis could be a major determinant of fitness for larvae of different genotypes responding to the stress of environmental change.

Shifting Balance of Protein Synthesis and Degradation Sets a Threshold for Larval Growth Under Environmental Stress.

Exogenous environmental factors alter growth rates, yet information remains scant on the biochemical mechanisms and energy trade-offs that underlie variability in the growth of marine invertebrates. Here we study the biochemical bases for differential growth and energy utilization (as adenosine triphosphate [ATP] equivalents) during larval growth of the bivalve Crassostrea gigas exposed to increasing levels of experimental ocean acidification (control, middle, and high pCO2, corresponding to ∼400, ∼800, and ∼1100 µatm, respectively). Elevated pCO2 hindered larval ability to accrete both shell and whole-body protein content. This negative impact was not due to an inability to synthesize protein per se, because size-specific rates of protein synthesis were upregulated at both middle and high pCO2 treatments by as much as 45% relative to control pCO2. Rather, protein degradation rates increased with increasing pCO2. At control pCO2, 89% of cellular energy (ATP equivalents) utilization was accounted for by just 2 processes in larvae, with protein synthesis accounting for 66% and sodium-potassium transport accounting for 23%. The energetic demand necessitated by elevated protein synthesis rates could be accommodated either by reallocating available energy from within the existing ATP pool or by increasing the production of total ATP. The former strategy was observed at middle pCO2, while the latter strategy was observed at high pCO2. Increased pCO2 also altered sodium-potassium transport, but with minimal impact on rates of ATP utilization relative to the impact observed for protein synthesis. Quantifying the actual energy costs and trade-offs for maintaining physiological homeostasis in response to stress will help to reveal the mechanisms of resilience thresholds to environmental change.

[Implantation of the irradiation stent system intoportal vein: studies on normal beagles].

Objective: To assess the feasibility and safety of the irradiation stent system in portal vein on normal beagles. Methods: A portal vein irradiation stent system was composed of an Iodine-125 seeds-carrier and a conventional stent.Twenty beagle dogs were randomly assigned to receive treatment with a conventional stent (5 beagle dogs) or an irradiation stent system (15 beagle dogs in three groups received 11.1, 22.2, 33.3 MBq radioactivity, n=5 in each dose group). Follow-up methods included blood biochemical test, color Doppler sonographyand CT scan at 0, 15, 30, 60, and 120 days after implantation of irradiation stent system.Pathological tissues were obtained from sacrificed beagle dogs on the 120th day. Results: The portal vein irradiation stent systems and the conventional stents were successfully deployed into the targeted portal vein segment in all beagles, none was dislodged during the deployment or the follow-up period.Differences of blood biochemical indexes and portal vein flow volume measured by color Doppler sonography were not significant (P>0.05). Stent systems were morphologically intact and patent.None of the peripheral organs had hemorrhage, necrosis or perforation.Pathological tissues revealed that the systems were surrounded by fibrous tissues and a few inflammatory cells, but with no significant differences in all groups. Conclusion: It indicates that portal veinirradiation stent system is safe in all dose groups, and it is feasible to design a special irradiation stent system for each patient according to the size of the portal vein tumor thrombus.

sPD-L1 Expression is Associated with Immunosuppression and Infectious Complications in Patients with Acute Pancreatitis.

Acute pancreatitis (AP) with infectious complications has high mortality because of early-stage immunosuppression. The programmed cell death-1 (PD-1)/programmed cell death ligand 1 (PD-L1) pathway is an important host immunosuppression mechanism. Soluble PD-L1 (sPD-L1) expression regulates co-inhibitory signals in malignancies or autoimmune disorders; however, its effects in AP are unknown. Here, we evaluated whether serum sPD-L1 is involved in immune dysfunction and assessed its relationship with infectious complications in early AP. Blood samples were obtained from 56 patients with acute pancreatitis and 21 healthy individuals in this prospective study. Serum sPD-L1 levels within 48 h after AP onset were tested by enzyme-linked immunosorbent assays. Relevant immune parameters (human leucocyte antigen-DR, lymphocyte count) and inflammatory markers (C-reactive protein, white blood cell count) were analysed. sPD-L1 was significantly upregulated in patients with early AP, especially those with infectious complications, compared to healthy controls. Significant negative correlations were observed among monocyte HLA-DR expression, lymphocyte count and sPD-L1 levels in AP. Multivariate regression indicated that sPD-L1 was an independent risk factor for infectious complications in AP. The findings suggest that increased sPD-L1 expression appears to be involved in the development of immunosuppression in the early stage of AP and that sPD-L1 might be an early parameter for prediction of infectious complications in patients with AP.

[The pilot study of type â allergic reaction in Meniere's disease patients].

Objective: To evaluate the correlation between type Ⅰ allergic reaction and pathogenesis of Meniere's disease. Methods: A total of 35 (10 male vs. 25 female) patients aged between 21-66 years diagnosed with Meniere's disease were recruited to this study, mean age of them was (47.3±13.6) years. The control group consisted of 15 inpatients (5 male vs. 10 female) with pharyngolaryngeal diseases but without otologic and rhinologic abnormity, mean age was 45.4±12.8 years. Allergic prevalence, serous total immunoglobulin E( tIgE ) levels, serous specific immunoglobulin E( sIgE ) levels and subtypes of T lymphocytes were measured and compared in patients with Meniere's disease and the control group. Severity of vertigo, tinnitus and sensation of fullness were compared between Meniere's disease patients with or without allergy. Results: Allergic prevalence were significantly different (Pearson chi-square 5.832, P<0.05) between patients with Meniere's disease and the control group(57.1% vs. 20.0%). Patients with Meniere's disease report higher level of serous tIgE compared with controls, the difference is statistically significant (Z=168.000, P<0.05). However, positive rates of sIgE of food allergens and inhalant allergens were not significantly different between patients with Meniere's disease and the control group. Scores of vertiginous severity, dizziness handicap inventory (DHI) and tinnitus handicap inventory (THI) were significantly different between Meniere's disease patients with or without allergy (P<0.05). Treg and Treg/Th17 levels (Z=26.000) were much higher in Meniere's disease patients with allergy than in the controls(P<0.05). Conclusions: Patients with Meniere's disease report higher rate of allergy than the control group. Type Ⅰ allergic reaction is thought to be one of the possible reasons that may induce endolymphatic hydrops and lead to Meniere's disease.

Celastrol, an active constituent of the TCM plant Tripterygium wilfordii Hook.f., inhibits prostate cancer bone metastasis.

BACKGROUND: Treatment failure of prostate cancer (PCa) is often due to bone metastasis. Celastrol, an active constituent of Tripterygium wilfordii roots, has shown anti-tumor effects in previous studies in accordance with its indication in traditional Chinese medicine. METHODS: Using a PC-3 cell model, in vitro assays were performed to evaluate the effects of celastrol on proliferation, migration (wound healing assay), tissues invasion (Transwell-Matrigel penetration assay) and vascular endothelial growth factor (VEGF) secretion (enzyme-linked immunosorbent assay). An intra-tibia injection mouse model was used to assess the effect of celastrol on PCa bone metastasis in vivo. RESULTS: Pretreatment with celastrol significantly reduced proliferation of PC-3 cells in a dose-dependent manner and cell migration was much slower than in controls. Significantly fewer cells penetrated the gel-membrane after celastrol administration and their skeletal invasive ability was significantly reduced in a dose-dependent manner. Correspondingly, a significant, dose-dependent decrease in VEGF secretion was observed. In the in vivo mouse model, pretreatment with celastrol (8 µmol l-1) inhibited the tumorigenicity of PC-3 cells so that almost no bone invasion occurred as compared with control injections. Histological examinations using hematoxylin and eosin staining showed that tibiae injected with celastrol pretreated PC-3 cells retained their natural bone structure. CONCLUSIONS: Celastrol may have preventive potential against PCa bone metastasis.

[Bone conduction in tympanosclerosis patients and its changes after surgery for tympanosclerosis].

Objective:To investigate the bone conduction (BC) in tympanosclerosis patients and its changes after surgery for tympanosclerosis, as well as its possible causes.Method:Retrospectively analyzed 30 cases of tympanosclerosis patients.Result:Before surgery, the average BC of the operated ear was (21.06±8.88) dBHL, the average BC of the healthy side was (16.78±8.97) dBHL, the difference had a statistical significance (t= 3.072, P< 0.01). The BC threshold, air conduction (AC) and ABG were significantly lower after operation. Preoperative BC at 2 000 Hz was the highest threshold (25.17±10.95) dBHL, and the postoperative decrease of BC thresholds at 500 Hz, 1 000 Hz and 2 000 Hz were statistically significant (P< 0.05 or P< 0.01). Tympanoplasty with mastoidectomy for BC threshold improvement was better than only tympanoplasty (P< 0.05)Conclusion:Tympanosclerosis leads to elevated BC threshold. Removal of tympanosclerosis lesions together with the removal of the lesions in antrum and mastoid and the reconstruction of sound conduction structures not only improve AC hearing and shorten ABG, but also improve postoperative BC hearing significantly.