[Cytomegalovirus pneumonia after allogeneic hematopoietic stem cell transplantation].

To compare the clinical features and prognosis in patients with cytomegalovirus pneumonia from other pneumonia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). A total of 118 patients with pulmonary complications after allo-HSCT from March 2016 to June 2019 were analyzed retrospectively, who were divided into cytomegalovirus (CMV) pneumonia group (n=34) and the non-CMV pneumonia group (n=84). Compared with non-CMV pneumonia group, CMV pneumonia group presented earlier median onset time (1.8 vs.6.0 months, P=0.015) after allo-HSCT, more dyspnea (41.2% vs. 19.0%, P=0.012), hypoxemia (38.2% vs. 13.1%, P=0.006), and interstitial pneumonia (82.4% vs. 23.8%,P<0.01).The incidence of CMV-viremia and serum viral load in CMV pneumonia group were significantly higher than those in non-CMV pneumonia group. Consistently, and the development of mixed infection in CMV pneumonia group was higher than that of non-CMV pneumonia group (41.2% vs. 16.7%, P=0.013). The median follow-up time was 12.8 (0.4-46.5) months. The 1-year attributable mortality in CMV pneumonia group was significantly higher than that in non-CMV pneumonia group (26.5% vs. 10.7%, P=0.004), while the 1-year overall survival rate was significantly lower than that in non-CMV pneumonia group (61.8% vs. 85.7%, P=0.001). Reduced-intensity conditioning (RIC)(P=0.036), high flow ventilation (P=0.033) and negative CMV-viremia (P=0.009) were unfavorable prognostic factors of patients with CMV pneumonia. Compared with those with non-CMV pneumonia, patients with CMV pneumonia had more characteristic clinical manifestations and imaging features. However, due to the higher incidence of mixed infections, the causes of pneumonia need to be identified by bronchoscopic alveolar lavage. In conclusion, patients with CMV pneumonia have worse clinical outcome. RIC, high flow ventilation and negative CMV-viremia are adverse prognostic factors for CMV pneumonia.

[Clinical study of 34 patients with cytomegalovirus pneumonia after allogeneic hematopoietic stem cell transplantation].

Objective: To analyze the clinical features and prognosis of cytomegalovirus pneumonia after allogeneic hematopoietic stem cell transplantation(allo-HSCT). Methods: We reviewed the clinical features and laboratory data of cytomegalovirus pneumonia patients after allogeneic peripheral blood HSCT from March 1, 2016 to June 30, 2019 at the hematology department of the Shanghai general hospital and analyze the prognostic factors. Results: Of the 411 allo-HSCT patients, 34(8.3%)developed CMV pneumonia after transplantation, including 18 men and 16 women, with a median age of 32(8-62)y. Total 14 patients had acute myeloid leukemia, 10 had acute lymphoblastic leukemia, 5 had myelodysplastic syndrome, 3 had non-Hodgkin's lymphoma, and 2 had aplastic anemia. The median onset time for CMV pneumonia was 53(36-506)d after transplantation. The main symptoms were cough(26 cases, 76.5%), fever(23 cases, 67.6%), and shortness of breath(14 cases, 41.2%). Only 17.6%(6/34)patients had expectoration, and 2 cases(5.9%)had no obvious symptoms in the early stage, but were diagnosed on routine chest CT examination. Twenty-eight(82.4%)patients showed signs of typical interstitial pneumonia, such as lobular central nodule and diffuse ground glass opacity; 6(17.6%)patients showed atypical imaging changes of patch, nodule, and consolidation. Further, 26 patients(76.5%)were positive for CMV-DNA, and the copy number was lower than that of BALF[1.70×10(7)(5.44×10(5)-4.45×10(9))copies/L vs 1.45×10(8)(1.10×10(7)-1.10×10(11))copies/L, P=0.004]. Thirteen(38.24%)patients with CMV pneumonia had mixed infection with other lower respiratory tract pathogens(10 strains of fungi, 6 strains of bacteria, and 1 of adenoviruses). The median follow-up duration was 12.8(0.4-46.5)months. The OS rate was 58.82%. Age ≥ 40 y and high flow ventilation were independent risk factors for poor prognosis in CMV pneumonia patients(P=0.049, P=0.009). Conclusion: Bronchoscopic bronchoalveolar lavage fluid detection helps in improving the accuracy of the etiological diagnosis of CMV pneumonia after allo-HSCT. Age ≥ 40 y and high flow ventilation were independent risk factors for poor prognosis in patients with CMV pneumonia.

LncRNA GAS5 induces chondrocyte apoptosis by down-regulating miR-137.

OBJECTIVE: Long non-coding RNA (lncRNA) participates in the pathogenesis of human knee osteoarthritis (KOA). Growth arrest specificity 5 (GAS5) is a member lncRNA, but its role in pathological regulation of KOA is still unknown. This study aims to explore the mechanism of GAS5 in KOA on chondrocyte apoptosis and other pathological processes. PATIENTS AND METHODS: The serum and cartilage tissues were collected from 35 patients with KOA and 30 patients with traumatic amputation admitted to our hospital from April 2016 to April 2020. The expressions of GAS5 and miR-137 were detected and analyzed. Chondrocytes were extracted from cartilage tissues of KOA patients, and the genes were regulated by transfection. Then, the cells were detected, including apoptosis, apoptosis-related proteins (caspase-3, Bax/Bcl-2), and proliferation. The targeting relationship between GAS5 and miR-137 was verified. RESULTS: GAS5 was up-regulated in serum and cartilage tissues of KOA patients, and down-regulation of GAS5 could inhibit the apoptosis process of chondrocytes and promote proliferation. MiR-137 was down-regulated in samples of KOA patients and was negatively regulated by GAS5. GAS5 induced apoptosis of chondrocytes and inhibited its proliferation through targeted down-regulating miR-137. CONCLUSIONS: GAS5 is up-regulated in KOA serum, cartilage tissues and cells, and can induce chondrocyte apoptosis through down-regulating miR-137.

MiR-1301 promotes adipogenic and osteogenic differentiation of BMSCs by targeting Satb2.

OBJECTIVE: Bone marrow mesenchymal stem cells (BMSCs) have the ability to differentiate into several cell lines and are critical for skeletal microenvironment and bone development. MiR-1301 is involved in multiple pathological and physiological processes. However, miR-1301's role in BMSCs adipogenic and osteogenic differentiation remains unclear. MATERIALS AND METHODS: Rat BMSCs were isolated and randomly divided into control group, miR-1301 group, and miR-1301 siRNA group followed by analysis of the expression of miR-1301, Bax, Bcl-2, UNX2, and OPN, as well as FABP4 and PPARγ2 by Real Time-PCR. Cell proliferation was assessed by MTT assay and the relationship between miR-1301 and Satb2 was evaluated by the Dual-Luciferase reporter assay. Satb2 expression was detected by Western blot. RESULTS: The pcDNA-miR-1301 plasmid was transfected into BMSCs to upregulate the expression of miR-1301, which promoted cell proliferation, decreased Bax expression, and increased Bcl-2 expression and ALP activity. In addition, it also elevated the expression of RUNX2 and OPN and decreased the expression of FABP4, PPARγ2, and Satb2. Compared with the control group, the difference was statistically significant (p<0.05); Satb2 was the target gene of miR-1301. MiR-1301 siRNA transfected into BMSCs down-regulated miR-1301 expression, inhibited cell proliferation, increased Bax expression and decreased Bcl-2 expression and ALP activity. Meanwhile, miR-1301 siRNA also reduced RUNX2 and OPN expression and increased expression of FABP4, PPARγ2 and Satb2. The difference was statistically significant compared with control group (p<0.05). CONCLUSIONS: Regulation of miR-1301 expression in BMSCs can improve BMSCs proliferation and regulate their adipogenic and osteogenic differentiation by regulating Satb2.

[Application of bronchoalveolar lavage fluid in patients with pulmonary complications after allogeneic hematopoietic stem cell transplantation].

Objective: To evaluate the diagnostic value of bronchoalveolar lavage (BAL) for pulmonary complications in patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its safety. Methods: Patients with pulmonary complications after allo-HSCT underwent BAL. Microbiological smears, culture, PCR of CMV-DNA, EBV-DNA and TB-DNA, macro genomes new generation sequencing (mNGS) techniques were performed to detect pathogens in BAL fluid (BALF) . Results: A total of 73 allo-HSCT patients with 86 times of pulmonary complications enrolled this prospective study. They underwent 132 times of BAL procedures. The clinical diagnoses of 88.4% cases were made based on BALF analysis. Of them, 67 cases (77.9%) had infectious pulmonary complications, including 29 cases (33.7%) of fungal infection, 18 cases (20.9%) of mixed infection, 11 cases (12.8%) of viral infection and 9 cases (10.5%) of bacterial infection. The other 9 cases (10.5%) of non-infectious pulmonary complications included 8 cases (9.3%) of idiopathic pneumonia syndrome (IPS) and 1 case (1.2%) of pulmonary infiltration of lymphoma. The diagnoses of the remaining 10 cases (11.6%) were not determined. The platelet counts of 33 patients were less than 50×10(9)/L before BAL. None of them developed severe bleeding complications during or after BAL. Transient fever occurred in 10 patients after BAL. Blood cultures showed staphylococcal bacteremia in them and anti-infection therapies were effective. No life-threatening complications occurred in all of the patients during or after BAL. Conclusion: BALF analysis was informative for the diagnosis of pulmonary complication and safe for patients with pulmonary complications after allo-HSCT.

[Treatment of 30 elderly patients with advanced myeloid neoplasm by allogeneic hematopoietic stem cell transplantation].

Objective: To evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for elderly patients with advanced myeloid neoplasm. Methods: From September 2014 to September 2017, 30 consecutive hospitalized 50-plus-year-old myeloid neoplasm patients were retrospectively analyzed. At the time of transplantation, 6 patients reached complete remission and the others remained no remission after treatment. The donors were identical sibling (12), matched unrelated (6) and haploidentical family member (12), respectively. 18 patients received RIC while 12 patients received MAC conditioning regiments consisted of Busulfan, cytarabine, fludarabine or clarithromycin±TBI, respectively. Results: Five patients died early in the conditioning stage, 24 patients successfully engrafted. The median time of neutrophil engraftment was 14(10-18) d, whereas platelet engraftment was 15(10-19) d. Six cases (25%) experienced aGVHD grades Ⅱ, 8 cases (32%) cGVHD, including moderate to severe cGVHD in 2 cases (8%). Seven, 7 and 5 cases developed CMV viremia, pneumonia and herpeszoster, respectively after transplantation, but no patients died of infections. The median follow-up time of the patients was 7(0.5-38) months. Twenty-one patients were still alive. The estimated 2 years OS and LFS were 62.5% (95% CI 39.2%-85.8%) and 59.2% (95% CI 26.9%-91.5%), respectively. Univariate analysis showed that HCT-CI was the only factor influencing OS. Conclusion: Allogeneic hematopoietic stem cell transplantation could improve the survival of elderly patients with myeloid neoplasm.

[Outcomes of reduced-intensity conditioning allogeneic hematopoietic stem cell transplantation for 10 patients with myelofibrosis].

Objective: To evaluate the efficacy of reduced-intensity conditioning allogeneic hematopoietic stem cell transplantation (RIC-allo-HSCT) for patients with myelofibrosis (MF). Methods: The clinical data of 10 patients with myelofibrosis (MF) who underwent RIC-allo-HSCT. Results: Of all 10 patients, 6 were male and 4 women, with a median age of 28.5 (22-54). Using fludarabine/busulfan plus total body irradiation (FB+TBI) pretreatment scheme based. Hematopoiesis reconstitution was achieved in 9 patients (90%). The median time of neutrophil and platelet engraftment was 13.5 (10-22) day and 16.5 (13-40) day, respectively. Acute GVHD occurred in 4 cases while chronic GVHD in 5 cases. The prospective OS for 3 years was (90.0±8.5)% after a median follow-up time of 17 months. Transplant related mortality was 1 case. Conclusion: RIC-HSCT with FB+TBI is a feasible and effective alternative for MF patients.

Anti-thymocyte globulin could improve the outcome of allogeneic hematopoietic stem cell transplantation in patients with highly aggressive T-cell tumors.

The early experiment result in our hospital showed that anti-thymocyte globulin (ATG) inhibited the proliferation of lymphoid tumor cells in the T-cell tumors. We used the ATG as the part of the conditioning regimen and to evaluate the long-term anti-leukemia effect, the safety and complication in the patients with highly aggressive T-cell lymphomas. Twenty-three patients were enrolled into this study. At the time of transplant, six patients reached first or subsequent complete response, three patients had a partial remission and 14 patients had relapsed or primary refractory disease. The conditioning regimen consisted of ATG, total body irradiation, toposide and cyclophosphamide. The complete remission rate after transplant was 95.7%. At a median follow-up time of 25 months, 16 (69.6%) patients are alive and free from diseases, including nine patients in refractory and progressive disease. Seven patients died after transplant, five from relapse and two from treatment-related complications. The incidence of grades II-IV acute graft-vs-host disease (GvHD) was 39.1%. The maximum cumulative incidence of chronic GvHD was 30%. The most frequent and severe conditioning-related toxicities observed in 8 out of 23 patients were grades III/IV infections during cytopenia. Thus, ATG-based conditioning is a feasible and effective alternative for patients with highly aggressive T-cell tumors.

Comparative metabolism of 7H-dibenzo[c,g]carbazole and dibenz[a,j]acridine by mouse and rat liver microsomes.

The comparative metabolism of the carcinogenic pollutants 7H-dibenzo[c,g]-carbazole (DBC) and dibenz[a,j]acridine (DBA) was investigated in vitro using 3-methylcholanthrene (3MC) induced Sprague-Dawley rat and Hsd:ICR(Br) mouse liver microsomal preparations with benzo[a]pyrene (BaP) as the positive control. Metabolites were isolated and separated by HPLC and identified by spectroscopic and co-chromatographic techniques using synthetic standards. The major metabolites of DBC were the phenols: the 5-OH-DBC, 3-OH-DBC, and 2-OH-DBC. Traces of 1-OH-DBC were also found yet no dihydrodiols were identified. The major metabolites of DBA were the 3,4-diol-DBA and 5,6-diol-DBA, 1,2-diol-DBA, DBA-5,6-oxide and 4-OH-DBA. Treatment of both mice and rats with 3MC resulted in significant (P less than or equal to 0.05) increases relative to control in the microsomal metabolism of DBA to dihydrodiol and phenol metabolites, similar to that observed for BaP. 3MC-induced rat liver microsomes significantly (P less than or equal to 0.05) increased DBC metabolism relative to control microsomes whereas DBC metabolism was not increased with 3MC-induced mouse liver microsomes. These data indicate that different enzymatic pathways are involved in the metabolic activation of DBC in the Hsd:ICR(Br) mouse and Sprague-Dawley rat.