Sequential therapy of targeting CD19 and CD22 chimeric antigen receptor T-cell following autologous stem cell transplantation for renal diffuse large B-cell lymphoma with central nervous system recurrence: report of 1 case and review of literature / 白血病·淋巴瘤

Objective:To investigate the efficacy and safety of sequential therapy of targeting CD19 and CD22 chimeric antigen receptor T-cell (CAR-T) following autologous stem cell transplantation (ASCT) in treatment of renal diffuse large B-cell lymphoma (DLBCL) with central nervous system (CNS) recurrence.Methods:The clinical data of 1 renal DLBCL patient with CNS recurrence admitted to Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology in May 2019 were retrospectively analyzed. The patient received sequential therapy of targeting CD19 and CD22 CAR-T following ASCT. The relative indicators of the primary disease remission at 1, 3, 6, 12, 18 and 24 months after therapy were analyzed, and relevant literature was reviewed.Results:The male patient aged 23 years had CNS recurrence after 8 courses of R-CHOP chemotherapy and then he received sequential therapy of targeting CD19 and CD22 CAR-T following ASCT. During the process of treatment, this patient developed grade 1 cytokine release syndrome and his condition was well controlled after active treatment. The white blood cell and platetes were successfully implanted. CNS symptoms along with immature cells in cerebrospinal fluid disappeared completely. Liquid biopsy was used to dynamically monitor the residue disease of the patient and the duration of remission period lasted 26 months. This patient developed tuberculosis one year after treatment and recovered from anti-tuberculosis agents.Conclusions:Sequential therapy of targeting CD19 and CD22 CAR-T following ASCT provides a novel therapeutic approach for renal DLBCL with CNS recurrence. Especially for patients who are neither sensitive to conventional chemotherapy nor CAR-T therapy alone, this regimen may improve remission rate and survival.

Flavonoids from stem and leaf of Scutellaria baicalonsis Georgi inhibit PHF abnormality and regulatory mechanism of protein phosphatase in rats' brain induced by okadaic acid / 中国病理生理杂志

AIM:To investigate the effect of flavonoids from stem and leaf of Scutellaria baicalonsis Georgi(SSF)on paired helical filament(PHF)abnormality and the regulatory mechanism of protein phosphatase(PP)in rats' brain induced by okadaic acid(OA).METHODS:Male Sprague-Dawley(SD)rats were microinjected with OA(200 ng/kg)by the lateral ventricle to establish a memory impairment model.Morris water maze was used to screen the memory impairment model.The successful model rats were continuous intragastric infusion(ig)SSF for 36 days.The relative pro-tein expression of PHF,PP1,PP2A-Cα,PP2A-Cβ,PP2CA and PP2CB in the rat cerebral cortex and hippocampus were detected by Western blot.GinKgo biloba leaf flavonoids(GLF)were used as positive control drug.RESULTS:Compared with the sham-operated rats ,the relative protein expression of PHF in the cerebral cortex and hippocampus and PP 1 in cor-tex of model rats were significantly increased(P<0.01),and the protein expression of PP2A-Cα,PP2A-Cβin the cere-bral cortex and hippocampus and PP2CB in the hippocampus were decreased(P<0.05),while the relative protein expres-sion of PP2CA and PP2CB in the cortex were significantly increased(P<0.01).SSF reversed the abnormality in the pro-tein expression of PHF,PP2A-Cαand PP2A-Cβin rat cortex and hippocampus and PP1 in rat cortex induced by OA(P<0.01),which had no significant effect on the relative protein expression of PP 2CA and PP2CB.GLF also showed similar results to SSF.CONCLUSION:SSF significantly reduces the abnormal formation of PHF in rats ' brain induced by OA ,which may be related to the regulation of PP 1,PP2A-Cαand PP2A-Cβexpression,but not with PP2CA and PP2CB ex-pression.

Differentially expressed genes and signalling pathways are involved in mouse osteoblast-like MC3T3-E1 cells exposed to 17-β estradiol / 国际口腔科学杂志·英文版

Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation. In this study, complementary DNA (cDNA) microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells' response to 17-β estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha (α-MEM) cell culture supplemented with 17-β estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with 10⁻⁸ mol⋅L⁻¹ 17-β estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase (ALP) activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction (RT-PCR) to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5403 differentially expressed genes, of which 1996 genes were upregulated and 3407 genes were downregulated, 1553 different functional classifications were identified by gene ontology (GO) analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta (TGF-β)-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to α-MEM supplemented with 17-β estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.

Establishment of xenotransplantation model of human CN-AML with FLT3-ITD (mut) /NPM1 (-) in NOD/SCID mice / 华中科技大学学报（医学）（英德文版）

Patients with FLT3-ITD (mut) /NPM1 (-) cytogenetically normal acute myeloid leukemia (CN-AML), as high-risk molecular group in CN-AML, are associated with a worse prognosis than other CN-AML patients. It is beneficial to generate xenotransplantation model of FLT3-ITD (mut) /NPM1 (-) CN-AML to better understand the pathogenesis and therapeutic strategies of such AML subtype. The purpose of present study was to establish the xenotransplantation model in NOD/SCID mice with FLT3-ITD (mut) /NPM1 (-) CN-AML primary cells. The FLT3-ITD (mut) /NPM1 (-) CN-AML primary cells from 3 of 7 cases were successfully transplanted into NOD/SCID mice, and human CD45 positive cells were detected in the peripheral blood, spleen and bone marrow of mice by using flow cytometry. Infiltration of human leukemia cells in various organs of mice was observed by using immunohistochemistry. Gene analysis confirmed sustained FLT3/ITD mutation without NPM1 mutation in mice. By performing serial transplantation, it was found that characteristics of the leukemia cells in secondary and tertiary generation models remained unchanged. Moreover, in vivo cytarabine administration could extend survival of NOD/SCID mice, which was consistent with clinical observation. In conclusion, we successfully established xenotransplantation model of human FLT3-ITD (mut) /NPM1 (-) CN-AML in NOD/SCID mice. The model was able to present primary disease and suitable to evaluate the curative effects of new drugs or therapy strategies.

Establishment of xenotransplantation model of human CN-AML with FLT3-ITD (mut) /NPM1 (-) in NOD/SCID mice / 华中科技大学学报（医学）（英德文版）

Patients with FLT3-ITD (mut) /NPM1 (-) cytogenetically normal acute myeloid leukemia (CN-AML), as high-risk molecular group in CN-AML, are associated with a worse prognosis than other CN-AML patients. It is beneficial to generate xenotransplantation model of FLT3-ITD (mut) /NPM1 (-) CN-AML to better understand the pathogenesis and therapeutic strategies of such AML subtype. The purpose of present study was to establish the xenotransplantation model in NOD/SCID mice with FLT3-ITD (mut) /NPM1 (-) CN-AML primary cells. The FLT3-ITD (mut) /NPM1 (-) CN-AML primary cells from 3 of 7 cases were successfully transplanted into NOD/SCID mice, and human CD45 positive cells were detected in the peripheral blood, spleen and bone marrow of mice by using flow cytometry. Infiltration of human leukemia cells in various organs of mice was observed by using immunohistochemistry. Gene analysis confirmed sustained FLT3/ITD mutation without NPM1 mutation in mice. By performing serial transplantation, it was found that characteristics of the leukemia cells in secondary and tertiary generation models remained unchanged. Moreover, in vivo cytarabine administration could extend survival of NOD/SCID mice, which was consistent with clinical observation. In conclusion, we successfully established xenotransplantation model of human FLT3-ITD (mut) /NPM1 (-) CN-AML in NOD/SCID mice. The model was able to present primary disease and suitable to evaluate the curative effects of new drugs or therapy strategies.

Mutation of isocitrate dehydrogenase 2 (IDH2) gene in Chinese AML patients and its clinical significance / 中国实验血液学杂志

This study was purpose to analyze the frequency and of isocitrate dehydrogenase 2 (IDH2) gene mutation in acute myeloid leukemia (AML) and its clinic significance. The multiplex polymerase chain reaction (PCR) and sequencing were performed to screen 192 AML patients for exon 4 of the IDH2 gene. FLT3, NPM1, CEBPA, c-kit and WT1 mutations were also included in analysis. The results showed that IDH2 mutation was found in 14 (7.29%) of 192 patients. There were 9 AML patients with R140Q mutation, 1 patient with R140W mutation, and 1 patient with R172K mutation. IDH2 aberrations significantly more were detected in French-American-British (FAB) M5 (P < 0.005) than other types. There was no statistical difference in age, sex, WBC, platelet count, bone marrow blasts count, hemoglobin as compared with IDH2 wild-type. For immunotype analysis, IDH2 mutation patients were more likely to express CD34 and CD13, less CD36. IDH2 mutation combined with FLT3/ITD mutation was found in 7 cases, with CEBPA mutation in 4 cases, with NPM1 mutation in 4 cases, with Dnmt3a mutation in 5 cases, neither with c-kit, IDH1 or WT1 mutation for no one, which revealed a significant interaction between IDH2 mutation and the FLT3/ITD positive genotype, Dnmt3a mutated, and IDH1 wild-type. IDH2 mutation was detected in 5 (8.47%) of 59 CN-AML. There was no significant difference of IDH2 mutation incidence between the normal and abnormal karyotype. The CR rate was higher in IDH2 R140 mutated patients than wild-type ones, but there was no significant in the two group. It is concluded that the rate of IDH2 mutation is 7.29% in Chinese AML patients and 7.81% in CN-AML. IDH2 mutation is significantly associated with AML-M5, FLT3/ITD, Dnmt3a, IDH1 wild-type and fusion gene wild-type, but not with age, leucocyte and platelet counts in peripheral blood, karyotype, NPM1, CEBPA, c-kit or WT1 mutation. And IDH2 R140 mutation has no impact on CR rate.

Study of immunophenotype of acute myeloid leukemia patients with NPM1 gene mutation / 白血病·淋巴瘤

Objective Toinvestigatetheimmunophenotypiccharacteristicsofacutemyeloidleukemia(AML) patients with NPM1 mutation. Methods The immunophenotype of 237 newly diagnosed AML patients were detected by flow cytometry. Real-time quantitative PCR was employed to detect the NPM1 mutation. The immunophenotype was then compared between the NPM1 mutated and wild type patients. Results The incidence of NPM1 mutation was 19.0 ％ (45/237) in all AML patients.The NPM1 mutated patients had lower expression of CD34,CD117,HLA-DR,CD15 and CD19 than the wild type patients(all P＜0.05).For AML patients with normal karyotype,the incidence of NPM1 mutation was 37.7 ％ (40/106),and the NPM1 mutated patients had lower expression of CD34,HLA-DR,CD15 and CD7 than the wild type patients(all P＜0.05).The NPM1 mutated patients with normal karyotype had lower expression of CD34 HLA-DR and CD7 in M1 subtype(all P ＜ 0.05); lower expression of HLA-DR and higher expression of CD9 in M2 subtype (all P ＜ 0.05) ; and lower expression of CD117 in M5 subtype compared with wild type patients (P ＜0.05). Conclusion The immunophenotypic characteristics of AML patients are changed by NPM1 mutation. The changes of immunophenotype varied in different FAB subtypes.

Detection of isocitrate dehydrogenase 1 gene mutation in 205 AML patients and its clinical significance / 中国实验血液学杂志

This study was purposed to detect the mutation of isocitrate dehydrogenase 1 (IDH-1) gene in patients with acute myeloid leukemia (AML) and to explore its clinical significance. The genomic DNA was extracted from mononuclear cells (MNC) of bone marrow or peripheral blood in 205 adult AML patients, the exon 4 of IDH1 gene was amplified by PCR, then the sequencing and comparison were performed. The results showed that IDH1 mutation was detected in 9 (4.39%) of 205 AML patients. There were 6 cases of R132H mutation, 1 of R132L mutation, 1 of R132G mutation and 1 of R132S mutation. Significantly more IDH1 aberrations were detected in AML-M2 (P = 0.002) than other types. And the 9 patients with IDH1 mutation were characterized by low platelet count which was lower than patients with wild type IDH1 (P = 0.003). IDH1 mutation combined with FLT3/ITD mutation was found in 5 cases, c-kit mutation in 1, NPM1 mutation in 2, and IDH1 mutation with CEBPA or WT1 mutation was not found, which revealed a significant interaction between IDH1 mutation and the FLT3/ITD positive genotype or the CEBPA wild-type. IDH1 mutation were detected in 4 of 71 (5.63%) CN-AML. There was no significant difference of IDH1 mutation incidence between the normal and abnormal karyotypes. It is concluded that the rate of IDH1 mutation was 4.39% in Chinese AML patients. IDH1 mutation is significantly associated with AML-M2, lower platelet counts in peripheral blood, FLT3/ITD mutation and CEBPA wild-type, but not with age, white blood cell count in peripheral blood, karyotype, NPM1, c-kit or WT1 mutation.

Analysis of IKAROS6 expression in 73 patients with chronic myelogenous leukemia / 中国实验血液学杂志

This study was aimed to evaluate IKAROS6 expression in patients with chronic myelogenous leukemia (CML) and its clinical significance. cDNAs from 73 CML patients were amplified by PCR and sequenced for IKAROS expression to elucidate clinical characteristics in IKAROS6 positive patients. The results showed that there was no IKAROS6 gene expression in 8 healthy controls and 15 CML patients in chronic phase and accelerated phase, and 15 cases (35.71%) were IKAROS6 positive in lymphoblast crisis samples among 42 newly diagnosed CML; however, none was found in myeloblast crisis of 16 newly diagnosed CML. Among 42 lymphoblast crisis of CML, the complete remission (CR) rate of IKAROS6 expression positive patients reached 40% (6/15), which was obviously lower than that in IKAROS6 negative patients (85.19%, 23/27) (p < 0.01), IKAROS6 positive patients relapsed after CR for 15 (2 - 18) months with relapse rate 66.7% (4/6), which was higher than that in expressed wild type IKAROS gene patients (21.74%, 5/23) (p < 0.05). It is concluded that abnormal expression of IKAROS gene dominated by IKAROS6 isoforms can be detected in lymphoblast crisis samples of CML patients. Abnormal expression of IKAROS gene may be an important factor in lymphoblast crisis of CML. Therefore, detection of IKAROS gene expression may be important for target therapy and evaluation of clinical prognosis of CML patients.

Application of FICTION technique to the detection of genetic aberrations in multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the diagnostic value of FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms) technique, combining immunofluorescence and fluorescence in situ hybridization (FISH), to detect genetic aberrations in multiple myeloma (MM).</p><p><b>METHODS</b>Bone marrow samples were collected from 18 MM and 2 plasma cell leukemia (PCL) patients. Probes targeting IgH and MMSET were prepared using a Nick Translation Kit from Bacterial artificial chromosome (BAC) clones. The immunophenotyping was achieved via the CD138 tyramide signal amplification (TSA)-mediated immunofluorescence, followed by FISH with the prepared probes \[t(4;14), t(11;14), t(14;16)\] and the commercial deletion probes (13q and p53) to detect common genetic aberrations in MM.</p><p><b>RESULTS</b>All the 20 samples were assayed with the probes mentioned above, and revealed 4 cases with t(4;14), 6 with t(11;14), 1 with t(14;16), 3 with p53 deletion; and 8 with 13q deletion. The remaining 4 cases had none of the 5 aberrations.</p><p><b>CONCLUSION</b>FICTION technique facilitates the detection of genetic abnormalities of MM in situ; enhances both efficiency and sensitivity of positive detection, thus, could be used as the screening test of molecular diagnosis of MM to guide coming-up risk-adapted therapy and evaluate prognosis.</p>

Splenic autotransplantation combined with lower esophagus transaction in the treatment of hepatic cirrhosis induced portal hypertension / 中华外科杂志

<p><b>OBJECTIVE</b>To study the therapeutic effect of splenic autotransplantation combined with lower esophagus transaction anastomosis in the treatment of liver cirrhosis induced portal hypertension.</p><p><b>METHODS</b>Thirty-six patients admitted from January 2003 to December 2006 were randomly divided into splenic autotransplantation group undergoing splenic autotransplantation after splenectomy combined with lower esophagus transaction anastomosis, and splenectomy group only undergoing splenectomy combined with lower esophagus transaction anastomosis. The general conduction, splenic scanning, liver function, and the level of serum Tuftsin and IgM of each patient were observed before and after operation.</p><p><b>RESULTS</b>The levels of Tuftsin and IgM in splenic autotransplantation group were significant higher than that of splenectomy group 2 months after the operation, and the liver function showed no significant difference between these two groups. Splenic tissue was detected in the retroperitoneal space by 99mTc-DRBC 2 months after operation.</p><p><b>CONCLUSIONS</b>Splenic autotransplantation combined with lower esophagus transaction anastomosis is a safe and effective treatment strategy for patients with liver cirrhosis induced portal hypertension, and the spleen tissue transplanted into the retroperitoneal space can partially preserve the immune function.</p>

MicroRNA differential expression profile during differentiation of embryonic stem cells towards hepatocytes induced by sodium butyrate / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the expression profile of microRNAs during the course of embryonic stem cells differentiation towards hepatocytes induced by sodium butyrate.</p><p><b>METHODS</b>Total RNA was extracted from embryonic stem cells on day 0, 6, and 9 during cell differentiation, and microRNA was isolated from the total RNA. Microarray analysis of microRNA expression was performed to detect the different expression levels of microRNA among the indicated time points (day 0, 6, and 9).</p><p><b>RESULTS</b>Compared with the microRNA expression level on day 0 of cell differentiation, 17 different microRNAs exhibited higher expressions both on day 6 and day 9. Twenty-two and 27 microRNA demonstrated lower expressions on day 6 and day 9, respectively. Further analysis revealed that 15 microRNA among the above microRNAs with significant differential expression may keep close interation with histone deacetylase.</p><p><b>CONCLUSION</b>During the course of embryonic stem cells differentiation towards hepatocytes induced by sodium butyrate, histone deacetylase and its relevant microRNAs may play important roles in cell differentiation.</p>

Diagnosis and treatment of primary tumor of small intestine: a report of 58 cases / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To summarize the pathological classification, clinical symptom and experience in the diagnosis and treatment of primary tumor of small intestine.</p><p><b>METHODS</b>Data of 58 patients with primary tumor of small intestine pathologically confirmed from Oct. 1996 to Oct. 2006 were analyzed retrospectively.</p><p><b>RESULTS</b>Thirteen patient (22.4%) had primary benign tumors of small intestine and 45 patient (77.6%) had primary malignant tumors of small intestine. The major clinical signs of primary tumor of small intestine included hemorrhage(85%), abdomen pain(19%), abdomen mass and intestine obstruction(16%). Forty- eight patients (82.8%) were diagnosed by laparotomy of abdominal cavity and misdiagnosed preoperatively as other diseases.</p><p><b>CONCLUSIONS</b>Primary tumors of small intestine are difficult to be diagnosed preoperatively. CT scan, digital subtraction angiography and radionuclide imaging are helpful for the diagnosis. Laparotomy of abdominal cavity is the main choice for those patients with suspicious tumor of small intestine.</p>