Objective To investigate the inhibitory effect of hydroxy safflor yellow A ( HSYA ) on angiogenesis of H22 tumor-bearing mice and it's effects on the protein expression of MMP-3 .Methods After establishing the hep-atoma model for 24 h, the mice were randomly divided into control group , sorafenib group and HSYA group , the dose HSYA group received intraperitoneal injection at different dosages (1.125 and 2.25 mg/kg).The pathologi-cal changes were examined with HE staining , immunohistochemical staining and Western blot were applied to meas-ure the expression of angiogenesis related factor ( MMP-3 ) and we also detected the microvessel density with CD 34 . Results Compared with control group , the tumor cells proliferation and the new angiogenesis in HSYA group were suppressed .The expression of MMP-3 in HSYA group was significant reduced .Especially the dose of 2.25 mg/kg HSYA group ( P<0.01 ) , and tumor MVD-CD34 was also significantly reduced ( P<0.01 ) .But the effect is not better than sorafenib group .Conclusions HSYA may inhibit angiogenesis of tumor tissue in a certain concentration range and the anti-angiogenesis effect of HSYA may be related to inhibition of the protein expression of matrix met-alloproteinase-3 .

Objective To investigate the distribution and antibiotic resistance profile of clinical isolates in the First Hospital of Qiqihar during 2015.Methods Antimicrobial susceptibility test was carried out according to a unified protocol using automated system from January 1,2015 to December 31,2015.The results were analyzed with WHONET 5.6 software according to the 2014 breakpoints of Clinical and Laboratory Standards Institute.Results A total of 5 162 clinical isolates were collected,of which 28.1％ (1 450/5 162) were gram-positive cocci and 71.9％ (3 712/5 162) were gram-negative bacilli.About 36.5％ (255/698) ofS.aureus isolates and 81.4％ (180/221) of coagulase negative Staphylococcus isolates were resistant to methicillin.No S.aureus and coagulase negative Staphylococcus isolate were found resistant to vancomycin or linezolid.Enterococcus isolates showed low resistance to vancomycin and linezolid.One strain of E.faecium was found resistant to vancomycin.ESBLs were produced in 39.9％ (298/747) ofE.coli,26.1％ (294/1 127) ofKlebsiella spp.,and 15.6％ (12/77) ofP mirabilis strains.The Enterobacteriaceae strains were less resistant to imipenem,beta-lactam/beta-lactamase inhibitor combination and amikacin.About 36.6％ (163 / 445) of A.baumannii isolates and 1.8％ (13/715) of P.aeruginosa isolates were extensively drug-resistant strains.Conclusions Antibiotic resistance poses a serious threat to clinical practice,to which more attention should be paid.Clinical microbiology lab should make more efforts to provide better support to clinical therapy.

Objective To detect the regional genomic instability of B16 cells treated with 60Co γ-rays by a green fluorescence protein (GFP)-based genomic instability reporting system.Methods Three groups were employed as non-transfection group,vector control group and transfection group.The GFP-marked reporter construct pCMV-EGFP2XhoI for regional genomic instability was successfully transfected into B16 cells using liposome.B16 cells were selected by screening of G418 with a series of concentrations and limiting dilution cultures to yield a single colony.B16 cells with the genomic instability report system were then irradiated by 60Co γ-rays at doses of 0,2 and 4 Gy.The regional genomic instability of B16 cellswas quantified by counting the number of cells with GFP expression.Results B-16 cell strain steadilyexpressing the GFP-based genomic instability reporting system was established successfully.GFP-positiveB16 cells were observed at 1 d after irradiation with 60Co γ-rays at doses of 2 and 4 Gy.Positive correlations between fluorescence intensity and dose and fluorescence intensity and time were also observed.The positive expression rate of GFP followed the increased of dose (F =36.55,36.76,P ＜ 0.05) and time (t =-3.27,-3.16,-4.26,-6.11,-7.17,P ＜ 0.05),and differences between groups were significant.The positive expression rate of GFP increased significantly at 3 d,and maximum expression was observed at 5 d(2.46 ± 0.24 and 3.82 ± 0.35).The level was tending towards stability.Spontaneous GFP expression at a ratio of 1/600 000 was observed in 0 Gy group after 2 weeks of culture.Conclusions The regional genomic instability of B16 cells induced by 60Co γ-rays can be detected using a GFP-labelled genomic instability reporter system.

Objective: To study the value of unmethylated cytosine guanine dinucleotide oligodeoxynucleotide (DSP30) and IL-2 in the conventional cytogenetic (CA) detection of the chromosomal aberrations in chronic lymphocytic leukemia (CLL) . Methods: Bone marrow or peripheral blood cells of CLL patients were cultured with DSP30 plus IL-2 for 72 h, following which R-banding analysis was conducted. Fluorescence in situ hybridization (FISH) was performed in 85 patients. CA results were compared with data obtained by FISH. Results: Among 89 CLL patients, the success rate of chromosome analysis was 94.38% (84/89) . Clonal aberrations were detected in 51 patients (51/84, 60.71%) . Of them, 27 (27/51, 52.94%) were complex karyotype. Among 85 CLL patients tested by FISH, chromosomal abnormalities were detected in 74 (74/85, 87.06%) patients, of which 2 (2/74) patients were complex karyotypes, accounting for 2.70%. Of the 85 CLL patients examined by FISH, 50 had abnormal karyotype analysis, 30 had normal karyotype, 5 failed to have chromosome analysis. Among them, 25 cases showed clonal aberrations by FISH assay but normal by CA, and 4 cases were normal by FISH but displayed aberrations in chromosome analysis, and totally 78 (91.76%) cases with abnormality detected by the combination of the two methods. The frequency of 13q- abnormality detected by FISH was significantly higher than that by CA analysis (69.41%vs 16.67%, P<0.001) , while the frequency of 11q-,+12 and 17p- detected by two methods showed no significant difference (P>0.05) . The detection rate of complex abnormalities in conventional karyotype analysis was higher than that in FISH (50.98%vs 2.70%) . In addition, 11 low-risk and 9 intermediate-risk patients according to FISH results showed complex karyotype by cytogenetics, and were classified into high-risk cytogenetic subgroup. Conclusion DSP30 and IL-2 are effective in improving the detection rate of CA in CLL patients (60.71%) and CA is more effective to detect complex karyotype. However, FISH had a higher overall abnormality detection rate (87.06%) than CA, especially for 13q-. The combination of CA and FISH not only enhanced the detection rate of clonal aberrations to 91.76%, but also provided more precise prognosis stratification for CLL patients, thus to provide more information for clinical implication.

OBJECTIVETo report clinical and laboratory features of 4 cases of myeloid neoplasm with t (5;12) (q33;p13).

METHODSCytogenetic examination of bone marrow cells obtained from patients was performed by 24 h culture method. R banding technical was used for karyotype analysis. PDGFRβ gene rearrangement was detected by FISH using dual color break apart PDGFRβ probe. ETV6-PDGFRβ fusion genes were detected by multiple-reverse transcription polymerase chain reaction (RT-PCR). Direct sequencing analysis was performed on the PCR products in case 1. Immunophenotype analysis was carried out by flow cytometry. Four cases were treated with imatinib (IM) and followed up.

RESULTSThe diagnoses included 3 MPN and 1 AML-M2. The t (5;12) (q33;p13) was a primary abnormality in 3 cases of MPN and a secondary abnormality in 1 case of AML-M2. PDGFRβ gene rearrangement and ETV6-PDGFRβ fusion genes were detected by FISH and multiple-RT-PCR in 4 cases, respectively. The immunophenotypical analysis of leukemia cells showed positive for CD13, CD33 and CD34. Two cases obtained MMR after the treatment of IM, one case complete hematologic and complete cytogenetic response. ETV6-PDGFRβ was negative detected by multiple-RT-PCR after the treatment of IM, but relapsed and died soon in case 4.

CONCLUSIONSThe t (5;12) myeloid neoplasm was a subtype with unique features. The t (5;12) maybe a primary chromosome abnormality in MPN and a secondary in AML. MPN with t (5;12) could benefit from IM, but not for AML. Dual-FISH was a reliable tool for detecting PDGFRβ rearrangement.

Chromosome Banding ; Gene Rearrangement ; Hematologic Neoplasms ; genetics ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Karyotyping ; Myeloproliferative Disorders ; genetics ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-ets ; genetics ; Receptor, Platelet-Derived Growth Factor beta ; genetics ; Remission Induction ; Repressor Proteins ; genetics ; Translocation, Genetic

OBJECTIVETo report on clinical and laboratory features of myeloid neoplasms with double del(20q).

METHODSCytogenetic examination of bone marrow was performed on 13 cases of myeloid neophasms with double del(20q) after 24 hours of cell culture. R-banding was used to analyze the karyotypes. Interphase fluorescence in situ hybridization (FISH) was performed using dual-color probes for 20q11/20q12.

RESULTSDouble del(20q) was found to be the sole abnormality in 9 cases, double del(20q) and trisomy 9 was found in 1 case, trisomy del(20q) was found in 1 case, and sole del(20q) clone and double del(20q) clone were found to coexist in 2 cases. In 10 cases, interphase FISH showed one green and one red signal in cells with del(20q), which indicated deletion of both 20q11 and 20q12. Immunophenotyping of the leukemia cells showed positiveness for CD13 and/or CD33, CD117 in all 9 cases. Among these, co-expression of CD34 and/or HLA-DR was found in 6 cases, and coexpression of CD3 and CD7 was found in 1 case. Of the 13 cases, there were one AML-M6, nine MDS, one pure amegalokaryocye aplastic thrombocytopenia, one with normal morphology of bone marrow, and one undetermined due to dilution of the bone marrow by blood. Cytopenia were found in all cases. 9 of 13 cases died, and 4 survived with a median survival of 9 months.

CONCLUSIONDouble del(20q) is a rare but recurrent chromosomal abnormality derived from del(20q). It has unique clinical and laboratory features, and the prognosis is poor.

Aged ; Chromosome Banding ; methods ; Chromosome Deletion ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Neoplasms ; genetics

OBJECTIVE: To explore the clinical and laboratory characteristics of 5 patients with myeloid leukemia and t(12;22)(p13;q12). METHODS: Bone marrow cells were cultured for 24 h and analyzed by standard R-banding. Rearrangement of the MN1 gene was detected by fluorescence in situ hybridization (FISH) using dual color break-apart MN1 probes. MN1-ETV6 and ETV6-MN1 fusion genes were detected by reverse transcription polymerase chain reaction (RT-PCR). And the products were subjected to direct sequencing. RESULTS: Among the 5 patients, 2 had AML-M0, 2 had AML-M4, and 1 had CMML at the initial diagnosis. t(12;22)(p13;q12) was the primary abnormality among all patients. Rearrangements of MN1 gene were detected by FISH in all patients. MN1-ETV6 and ETV6-MN1 fusion genes were detected respectively in 4 and 3 patients. CONCLUSION t(12;22)(p13;q12) is a rare but recurrent chromosomal abnormality in myeloid leukemia, and is related to poor prognosis. allo-SCT is valuable for patients with t(12;22)(p13;q12).
Chromosome Banding ; Chromosomes, Human, Pair 12 ; Chromosomes, Human, Pair 22 ; Cytogenetics ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myeloid ; genetics ; Oncogene Proteins, Fusion ; Translocation, Genetic

Objective To investigate the effect of carrimycin on the biological function of pancreatic cancer cells. Methods Pancreatic cancer cell lines MIA PaCa-2, BxPC-3, Panc-1, and PATU 8988 were treated with carrimycin at concentrations of 0 (control group), 2, 4, 8, and 16 μmol/L for 24, 48, and 72 hours. MTT assay was used to measure cell viability; EdU cell proliferation assay was used to observe the effect of carrimycin on DNA replication of pancreatic cancer cells; colony formation assay was used to observe the effect of carrimycin on the proliferation of pancreatic cancer cells; flow cytometry was used to analyze the effect of carrimycin on the cell cycle of pancreatic cancer cells; wound healing assay was used to analyze the effect of carrimycin on the migration of pancreatic cancer cells; Western blot was used to measure the expression levels of the markers such as epithelial-mesenchymal transition (EMT) and cell cycle-dependent protein kinase inhibitor 1A (P21); immunofluorescence assay were used to measure the expression levels of EMT-related markers. An analysis of variance was used for comparison between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the control group, carrimycin significantly inhibited the proliferative activity of MIA PaCa-2, BxPC-3, Panc-1, and PATU 8988 cells in a concentration- and time-dependent manner (all P < 0.01); carrimycin at concentrations of 4, 8, and 16 μmol/L significantly reduced DNA replication in MIA PaCa-2 cells ( t =2.378, 4.984, and 18.970, all P < 0.05) and BxPC-3 cells ( t =4.879, 6.089, and 9.521, all P < 0.01); after treatment with carrimycin at concentrations of 4, 8, and 16 μmol/L, colony formation ability significantly decreased with the increase in drug concentration in MIA PaCa-2 cells ( t =5.889, 11.240, and 15.840, all P < 0.001) and BxPC-3 cells ( t =6.717, 15.800, and 18.850, all P < 0.001). After treatment with carrimycin at concentrations of 4, 8, and 16 μmol/L, there was a significant increase in the proportion of cells in G1 phase in MIA PaCa-2 cells ( t =9.071, 12.280, and 19.360, all P < 0.0001) and BxPC-3 cells ( t =3.061, 4.962, and 8.868, all P < 0.05), and there was a significant reduction in the proportion of cells in S phase in MIA PaCa-2 cells ( t =2.316, 4.165, and 5.562, all P < 0.05) and BxPC-3 cells ( t =2.424, 3.264, and 5.744, all P < 0.05). Western blot further demonstrated that compared with the control group, the expression level of the cell cycle-related protein P21 gradually increased with the increase in the concentration of carrimycin in MIA PaCa-2 cells ( t =5.437, 6.453, and 8.799, all P < 0.001) and BxPC-3 cells ( t =25.130, 44.750, and 52.960, all P < 0.000 1). Wound healing assay showed that after treatment for 12, 24, and 48 hours, carrimycin at concentrations of 0, 4, 8, and 16 μmol/L significantly reduced the lateral migration of MIA PaCa-2 cells (all P < 0.05) and BxPC-3 cells (all P < 0.05). Western blot showed that compared with the control group, carrimycin treatment at concentrations of 4, 8, and 16 μmol/L significantly upregulated the expression of the epithelial marker E-cadherin in MIA PaCa-2 cells ( t =2.388, 4.899, and 5.819, all P < 0.05) and BxPC-3 cells ( t =2.533, 5.836, and 6.774, all P < 0.05) and significantly downregulated the expression of the interstitial marker Snail in MIA PaCa-2 cells ( t =12.440, 14.830, and 16.800, all P < 0.000 1) and BxPC-3 cells ( t=5.039, 5.893, and 7.725, all P < 0.01), and it also significantly downregulated the expression of the interstitial marker Vimentin in MIA PaCa-2 cells ( t =3.105, 7.752, and 11.200, all P < 0.05) and BxPC-3 cells ( t =2.555, 4.883, and 9.153, all P < 0.05). Conclusion Carrimycin can effectively inhibit the proliferation, migration, and EMT process of pancreatic cancer cells, thereby exerting an antitumor biological activity.

【Objective】 To investigate the effect of autologous blood transfusion(ABT) drainage system and simple drainage(using drainage bags) on the prognosis of patients after lumbar surgery. 【Methods】 The patients admitted to the Department of Orthopedics of our hospital from August 2018 to September 2020 who underwent posterior open lumbar internal fixation and fusion were divided into two groups according to different drainage methods adopted after surgery: 50 patients were randomly selected from the patients who received postoperative ABT system for drainage as ABT group, and 50 patients were randomly selected from the patients who received postoperative drainage by drainage bag as simple drainage bgroup(the control group). The postoperative drainage volume, actual postoperative drainage, total dominant blood loss, total autologous blood transfusion volume, as well as the postoperative anemia indexes, infection indexes and albumin levels in d1, d3 and d7 of the 2 groups were retrospectively analyzed. 【Results】 The gender, age, operation duration and operation segment of the 2 groups were comparable (P>0.05), and preoperative Hb, Hct, ALB, WBC, NE%, intraoperative blood loss, intraoperative autologous blood transfusion volume and actual postoperative drainage volume were similar (P>0.05). There were no significant differences in Hb, Hct, ALB, WBC, NE% and CRP in postoperative d1, d3 and d7, as well as in preoperative and postoperative Hb, Hct and ALB, and in postoperative and preoperative WBC, NE% and CRP (P>0.05). The postoperative drainage volume (mL) and total dominant blood loss (mL) in ABT group and the control group were 554.40±176.82 vs 337.80±102.43, and 1 048.40±282.87 vs 791.80±277.02, respectively (P<0.05). 【Conclusion】 The use of ABT drainage system after lumbar surgery increased the drainage volume compared with simple drainage, but the improvement of anemia, albumin and infection was not obvious. ABT system should not be used routinely for drainage after lumbar surgery.