Immunostimulatory gene therapy combined with checkpoint blockade reshapes tumor microenvironment and enhances ovarian cancer immunotherapy

Immune evasion has made ovarian cancer notorious for its refractory features, making the development of immunotherapy highly appealing to ovarian cancer treatment. The immune-stimulating cytokine IL-12 exhibits excellent antitumor activities. However, IL-12 can induce IFN-γ release and subsequently upregulate PDL-1 expression on tumor cells. Therefore, the tumor-targeting folate-modified delivery system F-DPC is constructed for concurrent delivery of IL-12 encoding gene and small molecular PDL-1 inhibitor (iPDL-1) to reduce immune escape and boost anti-tumor immunity. The physicochemical characteristics, gene transfection efficiency of the F-DPC nanoparticles in ovarian cancer cells are analyzed. The immune-modulation effects of combination therapy on different immune cells are also studied. Results show that compared with non-folate-modified vector, folate-modified F-DPC can improve the targeting of ovarian cancer and enhance the transfection efficiency of pIL-12. The underlying anti-tumor mechanisms include the regulation of T cells proliferation and activation, NK activation, macrophage polarization and DC maturation. The F-DPC/pIL-12/iPDL-1 complexes have shown outstanding antitumor effects and low toxicity in peritoneal model of ovarian cancer in mice. Taken together, our work provides new insights into ovarian cancer immunotherapy. Novel F-DPC/pIL-12/iPDL-1 complexes are revealed to exert prominent anti-tumor effect by modulating tumor immune microenvironment and preventing immune escape and might be a promising treatment option for ovarian cancer treatment.

Suture guided gonioscopy-assisted transluminal trabeculotomy combined with phacoemulsification in the treatment of primary open angle glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the clinical efficacy of suture-guided gonioscopy-assisted transluminal trabeculotomy(GATT)combined with phacoemulsification in the treatment of primary open angle glaucoma(POAG).METHODS: A total of 84 patients(84 eyes)with POAG and cataract who underwent surgery in our hospital from January 2021 to July 2022 were selected and randomly divided into two groups. There were 43 cases(43 eyes)in the combined group who underwent suture-guided GATT combined with phacoemulsification, and 41 cases(41 eyes)in the simple group who underwent suture-guided GATT. The two groups were followed up for 3mo to compare the surgical success rate, intraocular pressure, topical intraocular pressure-lowering drugs, visual acuity and postoperative complications.RESULTS: There was no significant difference in overall success rate between the combination and simple groups at 3 mo after surgery(88% vs. 85%, P>0.05). The intraocular pressure levels and topical intraocular pressure-lowering drugs at 1wk, 1 and 3mo after surgery in the two groups were significantly lower than those before surgery(all P<0.05), but there was no significant difference between the two groups(all P>0.05). The visual acuity at 1wk, 1 and 3mo after surgery of patients in the combined group was significantly better than that in the simple group(P<0.01). During the follow-up period, the incidence of anterior chamber hemorrhage and transient hypertension in the combined group was significantly lower than that in the simple group(P<0.05).CONCLUSION: Both suture-guided GATT combined with cataract phacoemulsification and suture-guided GATT are effective treatment for POAG, however, suture-guided GATT combined with phacoemulsification has a lower incidence of anterior chamber hemorrhage and transient ocular hypertension.

Expression and Role of Integrin β-like 1 in Hepatocellular Carcinoma / 肿瘤防治研究

Objective To investigate the expression of ITGBL1 in hepatocellular carcinoma (HCC) and its role in promoting the proliferation, migration, and invasion of HCC cell lines. Methods RT-qPCR and immunohistochemistry were used in investigating ITGBL1 expression in 12 pairs of fresh HCC and adjacent normal liver tissue samples and 160 paraffin HCC specimens. The relationships of ITGBL1 expression level with clinicopathological parameters and prognosis were analyzed. HUH7 cell line with the stable overexpression of ITGBL1 and LM3 cell line with stable downregulated expression of ITGBL1 were constructed. The effects of ITGBL1 on the proliferation, migration, and invasion of HCC cells were examined by CCK8 assay, wound-healing assay, and Transwell invasion assay. Results The expression levels of ITGBL1 gene and protein in tumor tissues were higher than those in surrounding liver tissues. The high expression of ITGBL1 was correlated with serum AFP level, tumor capsular invasion, vascular invasion, tumor differentiation, and clinical stage (P < 0.05). The results of Kaplan-Meier analysis showed that patients with high expression of ITGBL1 had shorter DFS. In vitro, increased ITGBL1 expression promoted HCC cell proliferation, migration, and invasion, whereas ITGBL1 knockdown inhibited these processes. Conclusion ITGBL1 expression is highly expressed in HCC tissues, and ITGBL1 can increase the proliferation, migration, and invasion of HCC cells. However, the mechanism needs further study.

Prognostic Threshold of Neuroendocrine Differentiation in Gastric Carcinoma: a Clinicopathological Study of 945 Cases

PURPOSE: The significance of neuroendocrine differentiation (NED) in gastric carcinoma (GC) is controversial, leading to ambiguous concepts in traditional classifications. This study aimed to determine the prognostic threshold of meaningful NED in GC and clarify its unclear features in existing classifications. MATERIALS AND METHODS: Immunohistochemical staining for synaptophysin, chromogranin A, and neural cell adhesion molecule was performed for 945 GC specimens. Survival analysis was performed using the log-rank test and univariate/multivariate models with percentages of NED (PNED) and demographic and clinicopathological parameters. RESULTS: In total, 275 (29.1%) cases were immunoreactive to at least 1 neuroendocrine (NE) marker. GC-NED was more common in the upper third of the stomach. PNED, and Borrmann's classification and tumor, lymph node, metastasis stages were independent prognostic factors. The cutoff PNED was 10%, beyond which patients had significantly worse outcomes, although the risk did not increase with higher PNED. Tumors with ≥10% NED tended to manifest as Borrmann type III lesion with mixed/diffuse morphology and poorer histological differentiation; the NE components in this population mainly grew in insulae/nests, which differed from the predominant growth pattern (glandular/acinar) in GC with <10% NED. CONCLUSIONS: GC with ≥10% NED should be classified as a distinct subtype because of its worse prognosis, and more attention should be paid to the necessity of additional therapeutics for NE components.

Anti-inflammatory Actions of Heshi Yangshen Recipe on Rat Model of Nephrotic Syndrome / 广州中医药大学学报

Objective To explore the anti-inflammatory action of Heshi Yangshen Recipe on rat model of nephrotic syndrome induced by adriamycin. Methods Rat model of nephrotic syndrome was established by tail venous injection of one-dose of adriamycin. SD rats were divided into normal group(N = 10),model group(N =11), positive control group (N = 12), and high-, middle- , low-dose Chinese medicine groups (N = 12, respectively). Two weeks after modeling,rats in the normal group and model group were given gastric gavage of same volume of normal saline, rats in positive control group were given gastric gavage of benazepril aqueous solution at a dose of 10 mg·kg-1·d-1 in the former period of 15 days and at 10 mg·kg-1·d-1 in the later period of 20 days, and rats in high-,middle- ,low-dose groups were given gastric gavage of Heshi Yangshen Recipe at a dosage of 33.26, 16.63, 8.32 g·kg-1·d-1 respectively, for 30 continuous groups. During the treatment period, the general situation of rats was observed. After the completion of medication, the levels of 24-hour urinary protein, urine total proteins, urine creatinine were measured by biochemical analyzer, serum high-sensitivity C-reactive protein (Hs-CRP) level was detected by enzyme-linked immunosorbent assay(ELISA),and the expression levels of intercellular adhesion molecule 1 (ICAM-1)and vascular cell adhesion molecule-1 (VCAM-1)in rat renal tissue of various groups were determined by immunohistochemistry. Results The increase of body mass in all of the modeled rats was significantly slowed down (P < 0.01 as compared with the normal group). Compared with the normal group, the levels of 24-hour urinary protein, serum hs-CRP, and the expression of ICAM-1 and VCAM-1 in rat renal tissue were increased in the model group (P < 0.05 or P <0.01). Compared with the model group, the level of 24-hour urinary protein in the positive control group and high-dose Chinese medicine group was decreased (P < 0.05), and the levels of serum Hs-CRP, and the expression of ICAM-1 and VCAM-1 in rat renal tissue of the positive control group and high-,middle-,low-dose Chinese medicine groups were decreased (P < 0.05 or P < 0.01). Conclusion Heshi Yangshen Recipe exerts certain anti-inflammatory action, and its mechanism may be related with the decrease of ICAM-1 and VCAM-1 expression. High dosage of Heshi Yangshen Recipe has the strongest therapeutic effect.

The influence of HBV and its antigens on the expressions of JAK-STAT signal transduction pathway molecules and antiviral proteins of IFN alpha / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the possible influence of HBV and its antigens on the expressions of JAK-STAT signal transduction pathway molecules and the antiviral proteins of IFN alpha.</p><p><b>METHODS</b>The HepG2 cells were transfected with pSM2, pHBS2-S and pHBc-EGFP plasmids which express HBV whole particles or S-antigen, Pre-S antigen and core antigens. The infectious supernatant from HepG2.2.15 cells and the pured HBV proteins which contained the S, Pre-S antigens were used to treat the HepG2 cells. Northern blot and RT-PCR were applied to analyse the expressions of the antiviral proteins MxA, 2' -5' OAS, 9-27 and the JAK-STAT signal transduction pathway molecules STAT1 in HepG2 cells responded to the IFN alpha treatment.</p><p><b>RESULTS</b>The HepG2 cells transfected with pSM2, pHBS2-S and pHBc-EGFP plasmids could express whole HBV particles and HBsAg, Pre-S antigen and HBcAg. The quantitation of expressed HBV particles and antigens increased significantly during the course of transfection. Northern blot hybridization analysis indicated that the HepG2 cells expressed IFN alpha antiviral proteins MxA, 2' -5' OAS and 9-27. When transfected with pHBV-dimer, pHBS2-S, pHBc-EGFP plasmids, the IFN/A antiviral proteins MxA, 2' -5' OAS and 9-27 in transfected cells were reduced greatly as compared to the un-transfected HepG2 cells, and the expressed antiviral proteins decreased sharply with the development of transfection time. Furthermore, the expression of IFN alpha JAK-STAT signal transduction pathway molecule STAT1 was also inhibited with the expression of HBV particles and HBV antigens in transfected HepG2 cells.</p><p><b>CONCLUSIONS</b>The HBV and its antigens influence the expressions of IFN alpha JAK-STAT signal transduction pathway molecules and antiviral proteins in the hepatocellular models in vitro. It is indicated that HBV might possess the activity to antagonise or counteract the IFN alpha antiviral action.</p>

Based on the low-density cDNA Macroarray for screening of antiviral proteins of IFNa tissues / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen the gene expression profiles of IFN-alpha antiviral proteins based on a low-density cDNA Macroarray, and to explore the relationship between the expression of antiviral protein and the HBV replication.</p><p><b>METHODS</b>The HepG2 and HepG2.2.15 cells were treated with various concentrations of IFN-alpha (0 IU/ml, 100 IU/ml, 1000 IU/ml) of IFN-alpha for 6 h, and then the low-density cDNA Macroarray was used for analysing the expression profiles of antiviral genes and screening differential expressions of antiviral proteins. Meanwhile, the HepG2 cells were transiently transfected with HBV core protein-expressed plasmid pHBc-EGFP, and the expressions of antiviral proteins were analysed by RT-PCR assay. Moreover, the HepG2.2.15 cells were also transfected with the antiviral protein-expressed plasmid pcDNA3.1-Flag-MxA. ELISA was used for analysing the secreted HBV antigens, while dot blot and Southern blot were applied for analysing the extracellular HBV DNA and intracellular replicative intermediate HBV DNA in HepG2.2.15 cells. All data were presented as mean+/-SD and analyzed using the t-test and one-way analysis of variance (ANOVA) in the experiments.</p><p><b>RESULTS</b>The Macroarray results suggested that the expression of IFN-alpha antiviral genes like 6-16, IFITM1, IFITM2, IFITM3 and RING4 in HepG2.2.15 cells were partially inhibited. More importantly, it was found, in this research, the expression of antiviral protein MxA in HepG2.2.15 cells was completely suppressed. RT-PCR analysis indicated that the expression of MxA was also significantly decreased in HepG2 cells transfected with pHBc-EGFP plasmid. Although HepG2.2.15 cells transfected with pcDNA3.1-Flag-MxA plasmid could not inhibit extracellular HBV DNA and intracellular replicative intermediate HBV DNA, the MxA exerted some antiviral activities as it effectively suppressed the secretion of HBsAg and HBeAg in HepG2.2.15 cells.</p><p><b>CONCLUSIONS</b>HBV and its antigen components probably influence the expression of antiviral proteins. IFN- resistance may be related to the down-regulation of antiviral proteins expression.</p>

Study on norcantharidin-induced apoptosis in SMMC-7721 cells through mitochondrial pathways / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of norcantharidin (NCTD)-induced SMMC-7721 hepatoma cell apoptosis.</p><p><b>METHODS</b>SMMC-7721 cell growth inhibition was measured by the MTT method. Apoptosis was detected by Annexin V/propidium iodide staining. The mitochondrial membrane potential was measured by flow cytometry. Western blot analysis was used to evaluate the level of cytochrome c, caspase-3, AIF, Bcl-2 and Bax expression.</p><p><b>RESULTS</b>NCTD inhibited SMMC-7721 cell growth in a time- and dose-dependent manner. The cells treated with NCTD showed the loss of mitochondrial membrane potential. The activities of caspase-3, cytochrome c, AIF, and Bax were up-regulated after NCTD treatment at different doses. The expression of Bcl-2 was decreased after treatment with NCTD.</p><p><b>CONCLUSIONS</b>NCTD could induce SMMC-7721 cell apoptosis. The activation of the mitochondrial pathway was involved in the process of NCTD-induced SMMC-7721 cell apoptosis.</p>

Effects of low-level lead exposure on the neurobehavioral development of infants and early intervention / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the effects of low-level lead exposure on infant's neurobehavioral development and evaluate the effects of early intervention.</p><p><b>METHODS</b>The study population consisted of 276 infants whose blood lead, cadmium, iron, zinc, copper, magnesium and calcium concentrations were measured by atomic absorption spectroscopy and developmental status were assessed using the Gesell developmental Diagnosis scales (GDDS) at 6 months of age. All study subjects was divided into three groups: 58 infants in control group, 162 infants in low lead group and 56 infants in high lead group. On the basis infants of both the low and high lead groups were provided with interventional measures for 3 months, and tests for the blood lead, cadmium, iron, zinc, copper, magnesium, calcium and GDDS were repeated for all infants both 12 and 18 months of ages.</p><p><b>RESULTS</b>Infant' s developmental outcome revealed the developmental quotient was the lowest in the high lead group (86.74 +/- 9. 35), the lesser low in the low lead group (91.52 +/- 10.12) and the highest in control group (100.71 +/- 6.92). Changes in developmental quotient were detected in both the low and high lead groups with statistical significance (P < 0.05) after intervention measures adopted. However, the changes of developmental quotient were more remarkable in the low lead group and after the 18th month there was no statistical significance than control group (t = 1.721, P > 0.05) while the significant difference was found in between the high lead group and the control group (t = 23.495, P < 0.05).</p><p><b>CONCLUSION</b>Low-level lead exposure interfered infant's neurobehavioral development and early intervention might improve infant's developmental quotient.</p>

Gene expression profile related to inflammation in rat model of traumatic deep vein thrombosis / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To study the relationship between inflammation and traumatic deep vein thrombosis (TDVT).</p><p><b>METHODS</b>A rat model of deep venous thrombosis was established by directly clamping femoral vein. Based on the different biological situations of femoral vein thrombosis and observation phases, 150 SD rats were divided into 7 groups. Inflammatory cells in vein wall of each group were counted. The fold change and cluster analysis were applied to study the change of gene expression during the development of venous thrombosis. Especially, the genes related to inflammation, fibrinolysis, coagulation of endothelium were analyzed in detail.</p><p><b>RESULTS</b>The inflammation cells in femoral vein wall were mostly neutrophilic granulocytes in Groups B, C and D, while they were lymphocytes in Groups E, F and G. Compared with Groups A, B, E and G, the inflammation cell counts in Groups C, D and F were much higher (P less than 0.05). The results of fold-change analysis showed that 2 504 genes (Log 2 ratio > or = 1 or < or = 1) presented different expressions in the process of TDVT. Most of these genes'functions were not clarified so far and the genes with known functions were involved in inflammation, DNA-dependent transcription regulation, blood coagulation, fibrinolysis, etc. Among them, 23 genes related to inflammation had different expressions during TDVT. The cluster analysis showed that the expression changes of several genes, such as IL-1 alpha, IL-1 beta, IL-6, Cinc2, corresponded with the development of femoral vein thrombosis.</p><p><b>CONCLUSION</b>There is a close relationship between the genes related to inflammation and deep vein thrombosis induced by direct vascular trauma.</p>

Significance of novel HBV expression vectors in selecting antiviral drugs in clinical therapy / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish a new method for rapidly selecting anti-hepatitis B virus drugs in clinical therapy.</p><p><b>METHODS</b>The full-length hepatitis B virus (HBV) genomes from 8 patients with chronic hepatitis B (CHB) were generated by polymerase chain reaction (PCR). All patients were resistant to lamivudine therapy. Their HBV DNA fragments were inserted into Sap I site of pHY106 eukaryotic expression vector separately. The recombinant plasmids containing 1.1 copies of HBV genome were transfected into Huh7 cell line; the levels of HBsAg, HBeAg and HBV DNA in supernatants of Huh7 cells were measured by ELISA and real-time quantitative PCR, and intracellular HBV replicative intermediates were detected by Southern blot. Antiviral effects of lamivudine and adefovir were evaluated in this vitro system.</p><p><b>RESULTS</b>The 8 recombinant plasmids containing a full-length genome of clinical HBV isolates could replicate and be expressed in Huh 7 cells. There were 6 isolates with polymerase YVDD mutations and 2 isolates with polymerase YIDD mutations. Adefovir, but not lamivudine, inhibited the HBV replication and gene expression in vitro. Furthermore, adefovir inhibited HBV replication in these CHB patients.</p><p><b>CONCLUSION</b>The method described here enables a rapid selection of anti-HBV drugs in clinical therapy and is very useful in antiviral therapy for CHB patients.</p>

Clinical and laboratory investigation of hematological malignancy patients carrying 3q21q26 rearrangement / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and laboratory characteristics of various hematopoietic malignant patients with t(3;3)(q21;q26) or inv(3) (q21q26).</p><p><b>METHODS</b>Bone marrow samples were collected at presentation, prepared by short-time unstimulated culture and R-binding, and karyotyped by conventional cytogenetical assay (CCA); megalokaryocytes were detected by Streptavidin-AKP (SAP); immunotype of the leukemia cells was tested by flow cytometric anylysis of surface antigens (FACS).</p><p><b>RESULTS</b>All of the 9 hematopoietic malignant patients with t(3;3)(q21;q26) or inv(3) (q21q26) manifested myelodysplasia and poor treatment response. One of them relapsed shortly after allogenic hemotopoietic stem cell transplantation (allo-HSCT).</p><p><b>CONCLUSION</b>Patients with 3q21q26 rearrangement can be found in various hematopoietic malignances and demonstrate an unique entity. These patients show poor treatment response and have extremely poor prognosis.</p>

Construction and Characterization of a Hepatitis B Virus Replicon / 中国病毒学

To establish a replication cellular model of hepatitis B virus (HBV) and determine its application in antiviral drug evaluation,we constructed an expression plasmid which contained 1.3 copies of the HBV genome,and measured the level of viral replication after transient transfection in Huh7 cells.We then observed the effect of antiviral drug administration.1.3 fold of the HBV(ayw) gene fragment was cloned into pCR2.1 by PCR and restriction endonuclease digestion.The recombinant plasmid was trans ient transfected into Huh7 cells,HBsAg,HBeAg and HBV DNA in supernatant of Huh7 cells were measured by ELISA and real-time PCR respectively; intracellular HBV replicative intermediates and intracellular HBV transcripts were detected by Southern blot and Northern blot respectively.The antiviral effect of adefovir,a novel anti-HBV nucleotide analogue,was evaluated in this cellular model system.The results indicated that a recombinant plasmid of HBV replicon was constructed successfully; the HBV genome carried in plasmid pHBV1.3 could efficiently replicate and be expressed in Huh 7 cells,adefovir could inhibit HBV replication in this cellular model,and the inhibition was dosage-dependent.The conclusion is HBV replicon,which can initiate viral replication efficiently in hepatoma cells,may be a useful tool in the study of HBV replication and antiviral drug.

Antiviral Effect of Interferon-Induced Guanylate Binding Protein-1 against Coxsackie Virus and Hepatitis B Virus B3 in Vitro / 中国病毒学

Guanylate binding protein-1(GBP-1) is an interferon-induced protein. To observe its antiviral effect against Hepatitis B virus (HBV) and Coxsackie virus B3 (CVB3), we constructed an eukaryotic expression vector of human GBP-1(hGBP-1). Full-length encoding sequence of hGBP-1 was amplified by long chain RT-PCR and inserted into a pCR2.1 vector, then subcloned into a pCDNA3.1(-) vector. Recombinant hGBP-1 plasmids and pHBV1.3 carrying 1.3-fold genome of HBV were contransfected into HepG2 cells, and inhibition effect of hGBP-1 against HBV replication was observed. Hela cells transfected with recombinant hGBP-1 plasmids were challenged with CVB3, and viral yield in cultures were detected. The results indicated that recombinant eukaryotic expression plasmid of hGBP-1 was constructed successfully and the hGBP-1 gene carried in this plasmid could be efficiently expressed in HepG2 cells and Hela cells. hGBP-1 inhibit CVB3 but not HBV replication in vitro. These results demonstrate that hGBP-1 mediates an antiviral effect against CVB3 but not HBV and perhaps plays an important role in the interferon-mediated antiviral response against CVB3.

Type IV secretion system in Helicobacter pylori: a new insight into pathogenicity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.</p><p><b>DATA SOURCES</b>The data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.</p><p><b>STUDY SELECTION</b>Mainly original articles and critical reviews written by major pioneer investigators of this field were selected.</p><p><b>RESULTS</b>The research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.</p><p><b>CONCLUSIONS</b>T4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.</p>

The prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>Chromosomal karyotypes in seventy-one MDS patients' were analyzed quantitatively. Based on the number of abnormal metaphase in 20 counted metaphases, the patients were divided into three groups: no abnormal karyotypes, abnormal metaphases less than or equal to five, and that more than five. The leukemia transformation rate, death rate and survival time between these three groups were compared.</p><p><b>RESULTS</b>Forty-four cases (62.0%) had abnormal karyotypes. The incidences of abnormal karyotypes in RA, RCMD and RAEB were 76.9%, 55.8% and 75.0%, respectively, being no significant difference (P > 0.05). Among the abnormal karyotypes, complex abnormality with two or more abnormal karyotypes was most common and accounted for 47.7%. The frequencies of trisomy 8, monosomy 7 and del 20q were 18.2%, 4.5% and 4.5%, respectively. Other kinds of abnormal karyotypes totally accounted for 25%. There were 27 cases of group 1, 28 of group 2 and 16 of group 3. Eighteen cases (25.4%) transformed to acute leukemia. The incidences of leukemia transformation in group 1, 2 and 3 were 18.5%, 25% and 37.5%, and the death rates were 29.6%, 42.9% and 56.3%, respectively. The median survival times were 60, 47 and 24 months respectively.</p><p><b>CONCLUSION</b>The quantitative chromosome abnormality has prognostic value in MDS.</p>

Application of dual-color fluorescence in situ hybridization in acute myeloid leukemia with t (8; 21) / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the utilities of dual-color fluorescence in situ hybridization (FISH) in diagnosis and monitor of treatment in acute myeloid leukemia (AML) with t (8; 21).</p><p><b>METHODS</b>Seventy patients having FISH results were divided into two groups: untreated and treated group. Comparative analysis was performed between the results of conventional cytogenetic analysis (CCA) and FISH analysis, and in some of them, between FISH and reverse transcriptase polymerase chain reaction (RT-PCR) results. A successive FISH following R-banding was carried out in those with cytogenetic undetermined cases.</p><p><b>RESULTS</b>In untreated group, 30/42 cases of t (8; 21) AML were positive for AML1/ETO in FISH assay. Three cases were positive for AML/ETO by FISH although two of them lacked t (8; 21) by CCA and one negative for AML1/ETO by RT-PCR. Six cases with complex karyotype abnormalities were confirmed to be AML1/ETO positive by the successive R-banding and FISH assay, and the involved genes were clearly visualized in FISH image. In the treated group, there were 28 cases of t (8; 21) AML diagnosed. Three cases without t (8; 21) by CCA were positive by FISH. Two patients were detected relapse earlier by FISH.</p><p><b>CONCLUSION</b>The dual-color FISH technique is a much more sensitive and accurate approach to the diagnosis of t (8; 21) AML and minimal residual disease (MRD) monitoring. It can also provide precise mapping of fusion signals in complex karyotype.</p>

Clinical and laboratory study of a complex translocation t (6; 21; 8) (p22; q22; q22) in two patients with acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and laboratory characteristics of a complex translocation t (6; 21; 8) (p22; q22; q22) in two patients with acute myeloid leukemia.</p><p><b>METHODS</b>Bone marrow (BM) samples were collected at presentation, prepared by short-term (24 hours) unstimulated culture and R-binding, for conventional cytogenetic assay (CCA). The complex translocation was assayed by fluorescence in situ hybridization (FISH) with a dual-color AML1/ETO-specific probe. AML1/ETO chimeric transcript was detected by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>In both cases CCA demonstrated a complex translocation, t (6; 8; 21) (p22; q22; q22), which was confirmed by interphase and metaphase FISH and AML1/ETO fusion transcript was detected by RT-PCR. Both the two patients were diagnosed as AML-M(2), but with different immunophenotype and therapeutic outcome.</p><p><b>CONCLUSION</b>The t (6; 21; 8) (p22; q22; q22) is a rare variant of complex translocation of t (8; 21) (q22; q22). More such cases are needed for elucidating its clinical features and prognosis.</p>

Burden of abnormal hematopoietic clone in patients with myelodysplastic syndromes / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>The ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor (TNF), CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed.</p><p><b>RESULTS</b>The clone burden of MDS patients was 67.4% +/- 36.2%. MDS clone burden positively correlated with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin, and erythrocytes in high clone burden (> 50%) and low clone burden ( < or = 50%) groups were 7.78% +/- 5.51% and 3.45% +/- 3.34%, 56.06 +/- 14.28 g/L and 76.40 +/- 24.44 g/L, (1.82 +/- 0.48) x 10(12)/L and (2.32 +/- 0.66) x 10(12)/L, respectively (all P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (0.274 +/- 0.719) x 10(9)/L and (0.455 +/- 0.206) x 10(9)/L, respectively (P < 0.05). CD8+ T lymphocytes of MDS patients and normal controls were (0.240 +/- 0.150) x 10(9)/L and (0.305 +/- 0.145) x 10(9)/L, respectively. The serum level of interleukin-2 of MDS patients (6.29 +/- 3.58 ng/mL) was significantly higher than normal control (3.11 +/- 1.40 ng/mL, P < 0.05). The serum level of TNF of MDS patients and normal control group were 2.42 +/- 1.79 ng/mL and 1.68 +/- 0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients (1.90 +/- 0.52) than that in low clone burden patients (0.97 +/- 0.44, P < 0.05).</p><p><b>CONCLUSION</b>The quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.</p>

Combination of interphase- and metaphase-fluorescence in situ hybridization to identify 11q23/MLL abnormalities in acute leukemia patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore a rapid, sensitive and effective method for identifying 11 q23/MLL gene rearrangements and investigate the incidence and clinical features of adult acute leukemia (AL) patients with 11 q23/MLL abnormalities.</p><p><b>METHODS</b>Bone marrow samples from 112 adult AL patients were prepared by short-term (24 hours) unstimulated culture, and karyotyped by R-banding. The abnormal signals were screened by interphase- fluorescence in situ hybridization (FISH) with dual-color break-apart 11 q23/MLL-specific probe, and the 11 q23/MLL gene rearrangements were determined by metaphase-FISH.</p><p><b>RESULTS</b>Of the 112 patients,9 (8. 0%) with 11q23/MLL translocations were revealed by FISH, among which only 4 (3. 6% ) was revealed by CCA. Three patients were reported by CCA to have del( 11) ( q23) , while by FISH assay two of them were 11 q23/MLL translocation and one was true deletion of I lq23 telomeric terminus. Furthermore by FISH assay II q23/MLL translocations were identified in one each patient with normal karyotype, with 11 q + and without overt 11 q23 abnormality. Eight patients with MLL gene amplification including polysome, homogenous staining region (hsr) and double minute chromosome (dmin) were also disclosed by FISH. AL patients with 11 q23/MLL abnormalities were frequently diagnosed as pro-B acute lymphoblastic leukemia (pro-B ALL) ,acute monocytic leukemia (AMoL) or biphenotypic acute leukemia (BAL).</p><p><b>CONCLUSION</b>FISH with dual-color break-apart I q23/MLL -specific probe is a rapid and sensitive method to detect 11 q23/MLL abnormalities, as compared with CCA. FISH also effectively discloses translocations and amplifications involving 11 q23/MLL,and should be performed in patients diagnosed as pro-B ALL,AMoL or BAL, with CCA normal karyotype.</p>