TAFRO syndrome with fatigue and abdominal distension as the first symptom: A case report.

TAFRO syndrome is a peculiar and rare type of multi-centric Castleman's disease which contained a series of symptoms such as thrombocytopenia (T), anasarca (A), fever (F), reticulin fibrosis (R), and organomegaly (O). Here we provide a case of TAFRO syndrome with the manifestation of fatigue, abdominal distension, and low fever at primary diagnosis, characterized by multiple lymphadenopathy of superficial mediastinal and retroperitoneal lymph nodes, and it was finally confirmed by lymph node biopsy. The patient recovered speedy after receiving CHOP chemotherapy. In this case report, the patient has a history of dust-exposure and hepatitis B virus infection, which may be potentially related to the disease. In addition, this case suggests the importance of pathological biopsy of complete lymph nodes in diagnostic process.

Variation in spatial distance between the lumbar interlaminar window and intervertebral disc space during flexion-extension.

PURPOSE: Knowledge of interlaminar space is important for undertaking percutaneous endoscopic discectomy via an interlaminar approach (PED-IL). However, dynamic changes in the lumbar interlaminar space and the spatial relationship between the interlaminar space and intervertebral disc space (IDS) are not clear. The aim of this study was to anatomically clarify the changes in interlaminar space height (ILH) and variation in distance between the two spaces during flexion-extension of the lumbar spine in vitro. METHODS: First, we used a validated custom-made loading equipment to obtain neutral, flexion, and extension 3D models of eight lumbar specimens through 3D reconstruction software. Changes in ILH (ILH, IL-yH, IL-zH) and distances between the horizontal plane passing through the lowest edge of the lamina of the superior lumbar vertebrae and the horizontal plane passing through the lowest position of the trailing edge of the same-level IDS (DpLID) at L3/4, L4/5 and L5/S1 were examined on 3D lumbar models. RESULTS: We found that ILH was greater at L4/5 than at L3/4 and L5/S1 in the neutral position, but the difference was not significant. In the flexion position, ILH was significantly more than that in neutral and extension positions at L3/4, L4/5, and L5/S1. There were significantly more DpLID changes from neutral to flexion than that from neutral to extension at all levels (L3/4, L4/5, L5/S1). CONCLUSION: These findings demonstrated level-specific changes in ILH and DpLID during flexion-extension. The data may provide a better understanding of the spatial relationship between lumbar interlaminar space and IDS, and aid the development of segment-specific treatment for PED-IL.

Haploidentical Peripheral Stem Cell Transplantation for Young Patients with Severe Aplastic Anemia Using Post-Transplantation Cyclophosphamide and Methotrexate.

Severe aplastic anemia (SAA) is a serious bone marrow failure disorder that is often cured with hematopoietic stem cell transplantation (HSCT). The absence of a matched related donor is common, however, and thus novel approaches are needed to safely expand the donor pool to include alternative donors, especially haploidentical related donors, for patients with SAA. This study aimed to explore a novel approach to HSCT for patients with SAA without an available HLA-identical sibling or a matched unrelated donor, termed haploidentical peripheral blood stem cell transplantation (haplo-PBSCT), using a conditioning regimen comprising cyclophosphamide, busulfan, and fludarabine (CBF) and a graft-versus-host disease (GVHD) prophylaxis regimen with post-transplantation cyclophosphamide (PTCy), low-dose methotrexate (LD-MTX), and calcineurin inhibitors. This prospectively designed nonrandomized study included 29 patients with SAA who underwent haplo-PBSCT between November 2017 and May 2020. The median patient age was 17 years (range, 14 to 30 years), and the median time to neutrophil recovery was 13 days (range, 13 to 15 days). There was 1 primary graft failure (GF) in the group receiving PTCy at a dose of 50 mg/kg and no GFs in the group receiving PTCy at a dose of 100 mg/kg. The median duration of follow-up was 736 days (95% confidence interval, 512 to 879 days). The estimated 1-year overall survival and disease-free survival were 91.7 ± 5.7% and 89.7 ± 5.7%, respectively. Only 1 of the 27 patients developed grade II acute GVHD. Four patients developed limited and mild chronic GVHD, involving only the skin or/and oral mucosa. Haplo-PBSCT following CBF and followed by PTCy and LD-MTX represents a novel approach for safely expanding the donor pool to include alternative donors for young patients with SAA.

Autopsy and statistical evidence of disturbed hemostasis progress in COVID-19: medical records from 407 patients.

BACKGROUND: The progression of coagulation in COVID-19 patients with confirmed discharge status and the combination of autopsy with complete hemostasis parameters have not been well studied. OBJECTIVE: To clarify the thrombotic phenomena and hemostasis state in COVID-19 patients based on epidemiological statistics combining autopsy and statistical analysis. METHODS: Using autopsy results from 9 patients with COVID-19 pneumonia and the medical records of 407 patients, including 39 deceased patients whose discharge status was certain, time-sequential changes in 11 relevant indices within mild, severe and critical infection throughout hospitalization according to the Chinese National Health Commission (NHC) guidelines were evaluated. Statistical tools were applied to calculate the importance of 11 indices and the correlation between those indices and the severity of COVID-19. RESULTS: At the beginning of hospitalization, platelet (PLT) counts were significantly reduced in critically ill patients compared with severely or mildly ill patients. Blood glucose (GLU), prothrombin time (PT), activated partial thromboplastin time (APTT), and D-dimer levels in critical patients were increased compared with mild and severe patients during the entire admission period. The International Society on Thrombosis and Haemostasis (ISTH) disseminated intravascular coagulation (DIC) score was also high in critical patients. In the relatively late stage of nonsurvivors, the temporal changes in PLT count, PT, and D-dimer levels were significantly different from those in survivors. A random forest model indicated that the most important feature was PT followed by D-dimer, indicating their positive associations with disease severity. Autopsy of deceased patients fulfilling diagnostic criteria for DIC revealed microthromboses in multiple organs. CONCLUSIONS: Combining autopsy data, time-sequential changes and statistical methods to explore hemostasis-relevant indices among the different severities of the disease helps guide therapy and detect prognosis in COVID-19 infection.

Ruxolitinib in treatment of severe coronavirus disease 2019 (COVID-19): A multicenter, single-blind, randomized controlled trial.

BACKGROUND: Accumulating evidence proposed Janus-associated kinase (JAK) inhibitors as therapeutic targets warranting rapid investigation. OBJECTIVE: This study evaluated the efficacy and safety of ruxolitinib, a JAK1/2 inhibitor, for coronavirus disease 2019. METHODS: We conducted a prospective, multicenter, single-blind, randomized controlled phase II trial involving patients with severe coronavirus disease 2019. RESULTS: Forty-three patients were randomly assigned (1:1) to receive ruxolitinib plus standard-of-care treatment (22 patients) or placebo based on standard-of-care treatment (21 patients). After exclusion of 2 patients (1 ineligible, 1 consent withdrawn) from the ruxolitinib group, 20 patients in the intervention group and 21 patients in the control group were included in the study. Treatment with ruxolitinib plus standard-of-care was not associated with significantly accelerated clinical improvement in severe patients with coronavirus disease 2019, although ruxolitinib recipients had a numerically faster clinical improvement. Eighteen (90%) patients from the ruxolitinib group showed computed tomography improvement at day 14 compared with 13 (61.9%) patients from the control group (P = .0495). Three patients in the control group died of respiratory failure, with 14.3% overall mortality at day 28; no patients died in the ruxolitinib group. Ruxolitinib was well tolerated with low toxicities and no new safety signals. Levels of 7 cytokines were significantly decreased in the ruxolitinib group in comparison to the control group. CONCLUSIONS: Although no statistical difference was observed, ruxolitinib recipients had a numerically faster clinical improvement. Significant chest computed tomography improvement, a faster recovery from lymphopenia, and favorable side-effect profile in the ruxolitinib group were encouraging and informative to future trials to test efficacy of ruxolitinib in a larger population.

Activation of the Nuclear Erythroid 2-Related Factor 2 Antioxidant Responsive Element (Nrf2-ARE) Signaling Pathway Alleviates Acute Graft-Versus-Host Disease by Reducing Oxidative Stress and Inhibiting Infiltration of Inflammatory Cells in an Allogeneic Stem Cell Transplantation Mouse Model.

BACKGROUND Acute graft-versus-host disease (aGVHD) limits the wider application of hematopoietic stem cell transplantation (HSCT). We explored the relationship between the Nrf2-ARE signaling pathway and aGVHD and identified effective and efficient therapeutic targets for the prevention and management of aGVHD following HSCT. MATERIAL AND METHODS C57BL/6 and BALB/c mice were used to establish the aGVHD model. The bone marrow and spleen mononuclear cells were separated from the donor mice and injected into the caudal vein of recipient mice that had undergone total body irradiation (TBI, 8 Gy). Sulforaphane (SFN) was used to activate the Nrf2-ARE signaling pathway. RESULTS The long-term survival rate of the SFN group was higher than that of the control group (40% vs. 0%, p<0.05, n=10). There were worse pathological changes and a greater infiltration of inflammatory cells in the liver, small intestine, and lung tissues of the control group. Furthermore, the Nrf2, NQO1, and HO-1 mRNA and protein levels were higher in the small intestines of the SFN group than in the control group (p<0.05, n=4). CONCLUSIONS The Nrf2-ARE signaling pathway plays a vital role in preventing aGVHD in an HSCT mouse model by regulating the expression of the downstream antioxidant genes NQO1 and HO-1 and by inhibiting the local inflammatory reaction.

[Effect of cryopreservation on umbilical blood cells and its mechanism].

OBJECTIVE: To evaluate the effect of cryopreservation on clonogenic ability and apoptosis rate of mono-nuclear cells and CD34+ cells in umbilical blood (UB), and to choose the index to present the freezing injury and optimize the cryopreservation of UB. METHODS: The mono-nuclear cells (MNC) and CD34+ cells were separated from UB and frozen.After 30 days, they were thawed in warm water. Clonogenic capacity and clonogenic recovery before and after the cryopreservation was compared. We also used Annexin V-FITC-PI to investigate the apoptosis rate of the cells before and after the cryopreservation of these 2 types of cells. RESULTS: The number of colony forming unit-granulocyte/monocyte (CFU-GMs) was not changed after freezing and thawing in both MNCs and CD34+ cells, while the number of colony forming unit-granulocyte, erythrocyte, monocyte and megakaryocyte (CFU-GEMM) was obviously reduced after freezing in CD34+ cells. The 2 types of cryopreserved cells had certain degree of apoptosis before the cryopreservation. MNC-type cryopreservation increased the cells apoptosis a little, while CD34+-type cryopreservation increased more. CONCLUSION: The cells have certain degree of apoptosis before the cryopreservation. The freezing and thawing procedure does affect the early stage progenitor cells-CFU-GEMM in the CD34+- type cryopreserved cells in UB. The damage may be induced by the cell apoptosis.

[Effect of human recombinant PDCD5 protein on cell apoptosis of multiple myeloma KM3 cells induced by dexamethasone and its mechanism].

OBJECTIVE: To observe the effect of programmed cell death 5 (PDCD5) protein on the apoptosis of multiple myeloma KM3 cells induced by dexamethasone and to understand its mechanism. METHODS: The human recombinant PDCD5 (rhPDCD5) protein was added (alone of different concentrations or associated with dexamethasone) into KM3 cells. Cultured together for certain time, the cells were collected for the following experiments: (1)The effect of rhPDCD5 protein and dexamethasone on the apoptotic rate of KM3 cells was determined by flowcytometry (FCM) analysis after the cells were stained by Annexin V-FITC & PI (propidium iodide). (2)Caspase-3 activity of KM3 cells was evaluated by Western blot. (3)The expression of survivin protein in KM3 cells was detected by immunocytochemistry. RESULTS: The apoptotic rate of KM3 cells and the activity of caspase-3 increased significantly, and that treated with rhPDCD5 protein and dexamethasone was higher than that treated with rhPDCD5 protein only. The expression of survivin protein in the rhPDCD5 with dexemethas group was down-regulated, and with the concentration of rhPDCD5 and dexamethasone increasing, the changes was more obviously. CONCLUSION: PDCD5 protein can induce the apoptosis of KM3 cells, and accelerate the apoptosis of KM3 cells induced by dexamethasone. PDCD5 protein may reduce the expression of survivin protein and increase activation of caspase-3 to play its role in promoting apoptosis.

[Expression of PDCD5 in multiple myeloma and its relation with BCL-2].

OBJECTIVE: To determine the expression of apoptosis related gene PDCD5 in multiple myeloma (MM), and to analyze the relation between PDCD5 and BCL-2. METHODS: The expressions of PDCD5 and BCL-2 protein and mRNA were determined by immunohistochemical staining method, flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR) method in bone marrow mononuclear cells. We also analyzed the relation between PDCD5 and BCL-2. RESULTS: Immunohistochemical staining showed that PDCD5 protein positive cell percentage, staining intensity index (SII) of PDCD5 protein, BCL-2 protein positive cell percentage, and SII of BCL-2 protein were (34.75 +/- 6.49)%, (281.16 +/- 75.33), (29.97 +/- 5.57)%, and (224.94 +/- 57.72) in the MM group and (52.98 +/- 5.84)%, (462.84 +/- 39.77), (5.56 +/- 1.95)%, and (27.84 +/- 9.75) in the control group (all P < 0.05). Results of FCM showed that PDCD5 protein positive percentage and mean fluorescence intensity of PDCD5 were (78.11 +/- 21.63)% and (61.73 +/- 11.04) in the MM group and (89.46 +/- 9.98)% and (353.04 +/- 123.26) in the control group (all P < 0.05). RT-PCR showed that relative expression of PDCD5 and BCL-2 mRNA were (0.33 +/ -0.07) and (0.33 +/- 0.08) in the MM group and (0.53 +/- 0.05) and (0.12 +/- 0.02) in the control group (all P < 0.05). The positive cell percentage of PDCD5 and BCL-2 protein was negative correlation (r = -0.86, P < 0.05); the expression of PDCD5 and BCL-2 mRNA was the same status (r = -0.90, P < 0.05). CONCLUSION: The expressions of PDCD5 protein and mRNA in MM patients are down-regulated, but the expressions of BCL-2 protein and mRNA are up-regulated. The mRNA and protein expression of PDCD5 and BCL-2 has negative correlation.

[Screening the effective target sequences of laryngeal carcinoma related gene LCRG1].

OBJECTIVE: To screen the effective target sequences of laryngeal carcinoma related gene LCRG1 using RNAi. METHODS: PCR site mutation method was used to reconstruct pSuper vector. Five pairs of siRNA sequences designed by siRNA software were annealed and inserted into the reconstructed pSuper vector. The reconstructed pSuper 362,398,432,789,903,and pSuper vectors were transfected into Hela cell lines and selected with the appropriate drugs to get resistant and pool cells, respectively. The colonies were identified by RT-PCR or real-time RT-PCR analysis. The silence effects were observed by cloning formation analysis. RESULTS: pSuper vector was reconstructed to restore Bgl II restriction enzyme sites using PCR mutation. The RT-PCR or real-time RT-PCR Results of pool clones showed 362, 398, and 432 pool clones all had better effects of LCRG1 gene-silence, especially 362 pool clones. The expression level of LCRG1 mRNA of selected 362 group anti-puromycin clones A2 and A5 was decreased. The Results of clone forming efficiency revealed that the cellular proliferation in A2 of 362 group was significantly higher than that of the vector and control Hela cells (P<0.05). CONCLUSION: The reconstructed pSuper vector is successfully constructed. The 362 group has better gene silence and has 2 effective 362 group anti-clones, suggesting that methodology has important values in studYing the function and molecular mechanism of LCRG1.

[3 polymorphisms of gene GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness].

OBJECTIVE: To investigate the relationship among 3 polymorphisms of GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness(PTR). METHODS: The 26th exon, the 30th exon and the 21st intron of gene GP IIb in 110 individuals were amplified by polymerase chain reaction (PCR), and the PCR products were analyzed with single-strand conformation polymorphism(SSCP) and sequenced to investigate whether there was linkage among the polymorphisms of the gene. Human platelet antigen-3 (HPA-3) gene frequency was detected by Fok I enzyme in 147 patients with hematologic diseases, and was compared with that in 110 normal individuals. Forty-four patients who received apheresis platelet transfusion repeatedly were randomly divided into the HPA-3 homotype group and the control group. The antibodies of the platelet were detected after 3 times of platelet transfusion. RESULTS: There were polymorphisms of gene GP IIb in the 26th, 30th exon and the 21st intron, and the mutations were: T changed into G in 13,959 th of the 26th exon; C changed into T in 16,997 th of the 30th exon; the 9 bps deletion occurred in 11,996-12,004 th of the 21st intron. The 3 polymorphisms had synchronization in the individuals. The results of Fok I enzyme indicated that the frequency of HPA-3a was 83.6% (92/110)and 81.9%(119/147), and that of HPA-3b was 16.4%(18/110) and 19.1%(28/147) in the normal individuals and the patients respectively. There was no significant difference between the patients and normal individuals (P>0.05). After the platelet transfusion, the antibodies of all the cases of the homotype platelet transfusion were negative, but the antibodies in 2 cases of the control group were positive, and there was antibody to HPA-3a in one of the antibodies positive cases. CONCLUSION: (1)There is close linkage among the polymorphisms of gene GP IIb, which is T->G in 13 959 th of the 26th exon, C->T in 16,997 th of the 30th exon, and the 9 bps deletion in 11,996-12,004 th in the 21st intron. (2)The gene frequency of HPA-3a/3b is similar in the normal individuals and patients with hematologic diseases. (3) HPA-3 system may be one of the reasons for PTR in Chinese.

[In vitro differentiation into megakaryocytes and generation of platelets from CD34+ cells of umbilical cord blood].

OBJECTIVE: To induce hematopoietic progenitor/stem cells of umbilical cord blood to differentiate into mature megakaryocytes and platelets in vitro and to investigate the mechanism of production of platelets. METHODS: The CD34+ cells were sorted from umbilical cord blood by magnetic activated cell sorting (MACS) and then cultured in vitro with optimized medium to be differentiated into mature megakaryocytes and platelets. The cultured cells and the platelet-like particles were isolated from the culture and were checked by the fluorescence-activated cell sorter (FACS), immunohistochemistry assays, light microscope,electron microscope and platelet aggregation tests. RESULTS: The cultured megakaryocytes were detected with proplatelets and both the cultured cells and the platelet-sized particles were found to have the same structure with the normal megakaryocytes and platelets by light and electron microscope. The immunohistochemistry assays revealed the cultured cells expressed GP II b III a with a positivity of 95% which was a special antigen for platelets and megakaryocytes. Culture-derived platelet-sized particles aggregated in response to thrombin as the plasma derived-platelets did. The cultured platelets had the same positivity of CD41 as the platelets from platelet rich plasma. CONCLUSION: The hematopoietic progenitor/stem cells can be induced to differentiate into purified and mature megakaryocytes and platelets. It provides a practical way to study the mechanism of platelets production.

[Detection of anti-HCV in blood recipients with hematonosis].

OBJECTIVE: To observe hepatitis C virus (HCV) infection in blood recipients with hematonosis, and to investigate the significance of anti-HCV detection in the patients. METHODS: Anti-HCV was detected by enzyme-linked immunosorbent assay (ELISA) in 176 hematonosis patients before blood transfusion, the result of anti-HCV was compared with control (417 cases), and 95 blood recipients were followed up for 6-12 months after the transfusion. RESULTS: The positive rate of anti-HCV was 5.68% (10/176) in hematonosis patients before transfusion, higher than both in the control [0.72% (3/417), P < 0.01] and in the patients with general surgery [2.23% (21/942), P < 0.05], and no new infection case was found after the transfusion. CONCLUSION: HCV infectionin in blood recipients with hematonosis should be paid attention to, and the detection of anti-HCV is necessary for patients before blood transfusion.