Urinary tract lymphoma: a clinical analysis of 16 cases and review of literature / 白血病·淋巴瘤

Objective To analyze the clinical features, treatment outcomes and prognosis of patients with urinary tract lymphoma. Methods The clinical data of 16 patients in Tongji Hospital of Tongji University from January 2009 to April 2016 were collected and retrospectively analyzed. Results The median age of these patients was 68 years. The onset symptoms of 14 cases were related to urinary system and imaging studies of 10 cases showed masses in the urinary system. The onset regions of lymphoma included:4 cases were renal lymphoma, 5 cases were adrenal lymphoma, 5 cases were testicular lymphoma, 1 case was prostate lymphoma and 1 case was from urethral mouth. The histological type of 12 cases was diffuse large B-cell lymphoma and 10 patients were non-germinal center B cell-like (non-GCB) molecular profiling. Twelve cases belonged to Ann Arbor stages ⅢE- ⅣE, 10 cases had international prognostic index scores ≥3, and 7 cases had B symptoms. 10 patients were confirmed by surgery. Fourteen cases accepted rituximab-containing regimen chemotherapy. Five cases achieved complete response and 3 were partial response. Conclusions The clinical manifestations and imagine examination of patients with urinary tract lymphoma are lack of specificity. The clinical features are highly aggressive and most of the patients are diagnosed at advanced stage. The main histological type is diffuse large B-cell lymphoma and non-GCB molecular profiling. Treatment regimens include surgery combined with chemotherapy and radiotherapy. Earlier diagnosis and treatment may improve the survival of patients.

Treatment of Ph-adult acute lymphoblastic leukemia patients with Tongji-96 regimen / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and side effects of the consecutive chemotherapeutic protocol, Tongji-96, for adult patients with Philadelphia chromosome negative acute lymphoblastic leukemia (Ph-aALL).</p><p><b>METHODS</b>A retrospective analysis was conducted on 95 cases of Ph-aALL patients treated between January 2004 and December 2012 with Tongji-96 regimen in Tongji hospital, Shanghai.</p><p><b>RESULTS</b>Among these 95 patients, the overall complete remission (CR) rate was 92.6%, 7-year overall survival (OS) and event-free survival (EFS) rates were (39.3±5.9)% and (31.5±5.3)%, respectively, with the median survival of 28 months. Based on multivariable COX proportional hazards regression model analysis, patients with the poor karyotype and failed to achieve CR after first course induction therapy had a higher risk of mortality compared to those who had good or normal cytogenetics and achieved CR after 1 course of induction treatment [the risk ratios (RR) were 3.380 (95% CI 1.530-7.463, P=0.003) and 3.005 (95% CI 1.522-5.933, P=0.002)，respectively]. By means of Kaplan-Meier analysis and Log-rank test，patients aged less than 60 years and successively achieved CR after first induction therapy had more favorable 7-year OS and EFS rates. Patients with normal karyotype and hyperdiploidy had significantly higher 2-year OS and EFS rates compared with those with complex karyotype, t(4;11) translocation and other karyotypes.</p><p><b>CONCLUSION</b>Age (60 years as the cut-off)，treatment courses for achieving CR and cytogenetics were predictive factors for the prognosis of Ph-aALL from this retrospective study. As a comprehensive and sequential therapy protocol, Tongji-96 regimen was proved to obtain long-term survival, reduce risks for relapse and improve outcomes for part Ph-aALL patients.</p>

Silencing of survivin gene in Jeko-1 cell line with small interfering RNA / 中国实验血液学杂志

This study was aimed to investigate the impact of specific siRNA on survivin gene in transfected lymphoma cell line and provide experimental evidences for future treatment of mantle cell lymphoma. The small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into Jeko-1 that showed high survivin expression in mRNA level. The levels of survivin mRNA and protein expression were detected by quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot respectively. The apoptosis effect was examined by calculating the ratio of Annexin V-FITC/PI positive cells using flow cytometry. The inhibition of cell proliferation was assayed with CCK-8 reagent after transfection. The results showed that expression of survivin mRNA was markedly suppressed by the siRNA. The relative expression levels were 0.49 ± 0.03, 0.38 ± 0.02 and 0.17 ± 0.02 at time points of 24, 48 and 72 h respectively, compared with the control group; the inhibitive rates of cell proliferation were (31.2 ± 2.1)%, (43.3 ± 3.4)% and (52.6 ± 2.5)%; the apoptotic rates of cells were (6.3 ± 0.5)%, (13.5 ± 1.1)% and (23.6 ± 1.6)% respectively; survivin protein expression levels were gradually reduced. It is concluded that the siRNA targeting survivin down-regulates the expressions of survivin mRNA and protein evidently. The siRNA of survivin displays the potent ability to inhibit the proliferation of lymphoma cell line Jeko-1; survivin may become a potential molecular target for the therapy of lymphoma in the future.

The effect of apoptosis induced by IL-21 in SUDHL-4 cell line and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis effect of diffuse large B-cell lymphoma cell line (DLBCL) SUDHL-4 induced by IL-21 and its related mechanism.</p><p><b>METHODS</b>SUDHL-4 cells were treated with IL-21 at different concentration (1000 ng/ml, 100 ng/ml, 10 ng/ml, 1 ng/ml) for 24 h, 48 h, 72 h, respectively. The inhibitory rate of cell proliferation was detected by CCK-8 assay. The cell growth curves were drawn and half inhibitory concentration (IC(50)) values were calculated. The cell apoptosis were detected by flow cytometry (FCM), the expression of the caspase-9, caspase-3, cleaved caspase-3, Bcl-2, Bcl-XL, Bid, Bax and c-myc protein in SUDHL-4 cells treated with IL-21 by western blot, the mRNA expression of Bcl-2, Bcl-XL, Bid, Bax, c-myc by Survivin gene with RT-PCR.</p><p><b>RESULTS</b>IL-21 markedly inhibited SUDHL-4 cell growth in a time- and dose-dependent manner. The 48 hIC(50) was 140.9ng/ml; The FCM showed that the apoptosis proportion of SUDHL-4 cells treated with 100 ng/ml of IL-21 apoptosis (AnnexinV-FITC(+) positive cells) gradually increased (48 h: 19.7 ± 2.3%). The protein expression of caspase-9, caspase-3, Bcl-2 and Bcl-XL decreased in a time-dependent manner. The Bax and c-myc protein markedly increased, but the Bid protein level did not change. IL-21 up regulated c-myc and Bax gene expression, however down regulated Bcl-2 and BCL-XL gene expression, but the gene expression of Bid and Survivin hadn't been changed significantly.</p><p><b>CONCLUSIONS</b>IL-21 can inhibit proliferation and induce apoptosis of SUDHL-4 cell. The mechanism may involve in endogenous mitochondrial pathway mediated by the c-myc and the Bcl-2 genes.</p>

Study of hypermethylation of SOCS gene in typical myeloproliferative disease / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical role of hypermethylation of suppressor of cytokine signaling (SOCS) on typical myeloproliferative disease (MPD) patients and its mechanism.</p><p><b>METHODS</b>Methylation specific PCR was used to detect SOCS1, 2, 3 methylation, direct DNA sequencing was performed to detect JAK2V617F mutation, real-time fluorescence quantitative PCR were applied to evaluate transcriptional activity of SOCS1, 2, 3.</p><p><b>RESULTS</b>Among 100 MPD patients, hypermethylation of SOCS1 was detected in 27 (27%), hypermethylation of SOCS2 in 9 (9%), hypermethylation of SOCS3 in 34 (34%); JAK2V617F mutation in 64 (64%). Hypermethylation of SOCS1, 3 greatly inhibited gene expression compared with unmethylated ones (P < 0.05). Presence of JAK2V617F mutation markedly down-regulated SOCS1, 3 gene mRNA expression compared with wild JAK2V617F (P < 0.05).</p><p><b>CONCLUSION</b>Hypermethylation of SOCS1, 3 and JAK2V617F mutation exist in MPD, which inhibited SOCS1, 3 gene expression. SOCS hypermethylation and JAK2V617F mutation can activate JAK-STAT signaling pathways, these observations may provide a potential therapeutic direction.</p>

The distribution and scintigraphy of 99Tcm labeled NGR-interferon-alpha2a in tumor bearing mice / 中华核医学杂志

Objective To sythesize 99Tcm labeled asparagine-glycine-arginine (NGR)- interferon (INF)-α2a and investigate its biodistribution by scintigraphy in tumor bearing mice. Methods NGR-INFα2a was labeled with 99Tcm by a two-step method. Ethylenedicysteine (EC) and MDP were used as bifunctional and transferring chelating agents. The bioactivities of 99Tcm-NGR-IFN-EC-NGR-IFN-α2a, EC-NGRIFN-α2a and NGR-IFN-α2a were compared using least significant difference t-test. The hepatoma bearing mice models were established by subcutaneous injection of MHCC97-H cells. The mice were randomly divided into eight groups and 7.4 MBq 99Tcm-NGR-IFN-α2a was injected via the tail vein. The tissue uptake of the radiolabeled compound was measured as % ID/g. The scintigraphy was performed at 0.5, 1, 2, 4,6, 8, 12 and 24 h after injection. ROI were drawn around tumor and non-tumor tissue and the radioactivity ratio of T/NT was calculated. Results Both the labeling efficiency and radiochemical purity of 99Tcm-EC-NGR-IFN-α2a were more than 90%. The radiochemical purity was 71% after 24 h in saline. The bioactivity showed no significant difference among three compounds (t = 0.416, 0. 120 and 1. 300, all P >0.05). The tracer was mainly excreted through alimentary and urinary tract within 24 h after injection. The peak values of % ID/g in kidney, liver, interstinal tract and tumor were 41.5 ± 8.0_ (at 8 h), 31.3 ± 5.0(at 6 h), 36.0 ± 7.8 (at 6 h), 43.0 ± 4.8 (at 4 h), respectively. The tracer was cleared quickly from the blood and the highest T/NT ratio was 16.5. The optimal imaging time ranged from 4 to 8 h after injection. Conclusions The sythesis of 99Tcm-NGR-IFN-α2a is applicable and it may be used as a potential tumor imaging agent.

Effect of hypoxia on HIF-1α and its sumoylation in Jurkat cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of hypoxia on sumoylation of HIF-1α, as well as transcriptional activity and protein stability of HIF-1α in Jurkat cells, and explore its effect and significance on modulating the transcriptional activity of down stream target gene.</p><p><b>METHODS</b>CoCl(2) was used as a chemical inducer to simulate the hypoxia environment. Real-time fluorescence quantitative PCR and western blot were used to evaluate transcriptional activity and protein stability of HIF-1α, levels of SUMO-1 and SENP1 protein, and gene transcripts of VEGF, mdr1, mdr3, Mcl-1 and survivin respectively.</p><p><b>RESULTS</b>After 4 h, 8 h, 16 h, 24 h and 72 h after hypoxia induction, the gene transcripts of HIF-1α were 0.79 ± 0.19, 2.65 ± 2.05, 4.19 ± 4.72, 2.77 ± 3.37, 0.09 ± 0.05 and 0.69 ± 0.55-fold (P > 0.01) of that of 0h in Jurkat cells, respectively, while the protein stability increased first, then decreased (P < 0.01). SENP-1 protein up-regulated first, then down-regulated, and the SUMO-1 protein changed in an opposite trend. Excepting for survivin gene, transcriptional activities of VEGF, mdr1, mdr3, and Mcl-1 were affected by the stability of HIF-1α protein.</p><p><b>CONCLUSION</b>Hypoxia induces changes in SENP-1 expression, which increases the stability of HIF-1α by decreasing the sumoylation of HIF-1α and affects biological process by regulating the transcriptional activities of VEGF, mdr1, mdr3 and Mcl-1 gene.</p>

Effect of VEGFR1 gene silencing by shRNA on proliferation, migration and apoptosis of U937 line / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of lentivirus-mediated RNA interference targeting vascular endothelial growth factor receptor 1 (VEGFR1) gene on the proliferation, migration and apoptosis of leukemic cell line U937.</p><p><b>METHODS</b>Short hairpin RNAs (shRNA) targeting VEGFR-1 was synthesized and cloned into pRNAT-U6.2 lentiviral vector. The expression vectors were transfected into 293T cell line to produce packaged lentivirus. After infected with the packaged lentivirus, the expression of VEGFR-1 gene of U937 cells at mRNA and protein level was detected by real-time PCR and Western blot. VEGF production by the cells was determined by ELISA. Cell proliferation and survival under regular culture and in the presence of cytarabine (Ara-C) was determined by CCK-8 assay. Migration assays were performed by 5 µm pore transwell inserts.</p><p><b>RESULTS</b>The lentiviral shRNA vector targeting VEGFR-1 was successfully constructed and transfected into U937 cells. The shRNA vector effectively inhibited the expression of VEGFR-1 gene in U937 cell line at mRNA and protein levels. As compared to that of the control, the proliferation rate of U937-shVEGFR-1 cells reduced; The VEGF production and migrated cell number of U937-shVEGFR-1 cells decreased dramatically. After treated with Ara-C, the inhibition rate and apoptotic rate of U937-shVEGFR-1 cells increased significantly. The number of migrated cells in the KD group under regular culture and in the presence of VEGF was markedly lower than that in the NC group and CON group. Bevacizumab could decrease the number of migrated cells in the NC group and CON group, but could not in the KD group.</p><p><b>CONCLUSIONS</b>Lentivirus-mediated RNA interference targeting VEGFR1 gene reduces the proliferation, migration of U937 cell line and enhances its sensitivity to Ara-C.</p>

Comparison of the influence of Eperythrozoon infection on human and mouse erythrocytes / 中华传染病杂志

Objective To evaluate the influence of Eperythrozoon infection on human and mouse erythrocytes and to explore the pathogenesis of Eperythrozoonosis. Methods The specific gene fragment of Eperythrozoon was detected by polymerase chain reaction (PCR) from the venous blood samples of five patients infected with Eperythrozoon. The complement receptor type I (CD35) expression on erythrocytes of these five patients was determined by flow cytometry. Thereafter, the Eperythrozoons were purified from human samples and injected into mice through the tail veins. Blood smear microscopy, PCR and transmission electron microscopy were used to assure the successful infection. The hematological indicators of human and mice, such as red blood cell (RBC) count,hemoglobin (Hb) content, hematocrit and superoxide dismutase (SOD) were evaluated. All results were analyzed by t test. Results More than 80% of treated mice were confirmed to be infected with Eperythrozoon successfully. A fragment of 801 bp specific gene of Eperythrozoon was detected by PCR in samples from both infected patients and infected mice, which were not detected in samples from healthy control people or control mice. CD35 was highly expressed on the erythrocytes of infected patients, but not expressed on the erythrocytes of infected mice. Both RBC counts and Hb content dramatically decreased in infected patients and infected mice. Hematocrit and the activity of SOD also slightly decreased in infected patients and infected mice. Conclusions Eperythrozoon can spread between human and mice and destroy erythrocyte structure. Eperythrozoon can upregulate CD35 expression in human, but there is no CD35 expression in mice.

Effect of microRNA on proliferation of human tongue carcinoma Tca8113 cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of microRNA (miRNA) on proliferation of cultured human squamous cell carcinoma of tongue Tca8113 cells.</p><p><b>METHODS</b>The mimics or inhibitors of miRNA-31 or miRNA-139 were transfected into Tca8113 cells using liposome. Tca8113 cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay.</p><p><b>RESULTS</b>The absorbance (A) values of control group at 24 h, 48 h and 72 h were 0.125 +/- 0.002, 0.169 +/- 0.002 and 0.216 +/- 0.004, respectively. The mimics of miRNA-31 increased Tca8113 cell proliferation, with A values increasing to 0.136 +/- 0.001 (P < 0.001), 0.186 +/- 0.004 (P < 0.001) and 0.249 +/- 0.012 (P < 0.01), respectively. The inhibitors of miRNA-139 also increased A values to 0.148 +/- 0.002 (P < 0.001), 0.214 +/- 0.002 (P < 0.001) and 0.250 +/- 0.009 (P < 0.01), respectively. Contrast with these results, the inhibitors of miRNA-31 decreased Tca8113 cell proliferation, with A values decreasing to 0.145 +/- 0.001 and 0.155 +/- 0.011 (both of P < 0.001) at 48 h and 72 h, respectively. The mimics of miRNA-139 also decreased A to 0.135 +/- 0.001 and 0.170 +/- 0.009 (both of P < 0.001).</p><p><b>CONCLUSIONS</b>miRNA-31 and miRNA-139 play an important role in the carcinogenesis of human tongue carcinomas. It may become a new method for the treatment of tongue carcinomas by adjustment the activities of miRNA.</p>

The anti-proliferation and anti-migration dual effects of aloe-emodin on KB cells and its mechanism / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the anti-proliferation and anti-migration dual effects of aloe-emodin on KB cells and its mechanisms.</p><p><b>METHODS</b>KB cells were treated with 2.5, 5, 10, 20 and 40 micromol/L aloe-emodin. Crystal violet assay was used to determine the long-term growth inhibition of aloe-emodin on human oral cancer KB cells. Scratch wound-healing motility assay was used to measure the antimigration effect The protein kinase C alpha and c-myc expression changes in protein levels were detected by Western blotting.</p><p><b>RESULTS</b>A durable cell growth inhibitory effect of aloe-emodin on KB cells was found. Treatment of aloe-emodin resulted in the inhibition of cell migration. The protein kinase C alpha and c-myc in protein levels were decreased upon treatment with aloe-emodin compared with controls.</p><p><b>CONCLUSIONS</b>The anti-proliferation and anti-migration effects of aloe-emodin on KB cells are associated with the suppression of protein kinase C pathway.</p>

Correlation between expression of platelet membrane glycoproteins and hyperviscosity syndrome / 第二军医大学学报

Objective:To study the relationship between hyperviscosity syndrome(HVS) and platelet membrane glycoproteins(GP) sub group changes.Methods: CD62P,CD63,CD31 and CD41 were measured by flow cytometry(FCM). The hemorrheoloic parameters of HVS patients were compared with those of normal aged group and normal young group.Results:(1)The CD62P,CD63 and CD31 of aged and young HVS patients before therapy were obviously higher than those of the normal aged group and normal young group.(2)Combined measurement could improve the detecting positive rate of HVS patients up to 92% 100%, with the sensitivity being 95.3%.(3)Bamyl decreased CD62P,CD63 and CD31,which was obviously correlated with ?b and ?p. Conclusion:The alteration of platelet membrane GP positive expression may be a parameter for early diagnosis and clinical outcome observation.