Objective To investigate the effects of nimesulide, a selective cyclooxygenase-2 (COX-2) inhibitor, on human cholangiocarcinoma QBC939 cell line in vitro. Methods The effects of nimesulide on QBC939 cells were observed with the following techniques: the influence of nimesulide on the proliferation of QBC939 cells was determined by MTT assay; the apoptosis of QBC939 cells was viewed and measured by transmission electron microscopy and flow cytometry, respectively; the expressions of proliferation cell nuclear antigen (PCNA) and COX-2 of cholangiocarcinoma cells were detected by immunocytochemistry. Results Nimesulide inhibited the expressions of PCNA and COX-2 and the proliferation of cholangiocarcinoma QBC939 cells, whose effects intensified as the dose increased and time elongated. Flow cytometry showed that the apoptotic rates of QBC939 cells increased significantly as the dose of nimesulide increased. The typical morphologic features of apoptosis were also observed by transmission electron microscopy. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 cells in vitro by inducting cell apoptosis, which may be associated with the downregulation of COX-2 expression, and it also presents the features of dose and time dependents.

Objective To investigate the effect of a selective inhibitor of COX-2 nimesulide on growth and apoptosis of human cholangiocarcinoma cell line QBC939 in vitro. Methods MTT assay was used to determine the influence of nimesulide on the proliferation of QBC939 cells, apoptosis of QBC939 was measured by transmission electron microscopy and flow cytometry.Expression of apoptosis related genes mRNA and bcl-2 ,bax, survivin were detected by RT-PCR and immunocytochemistry. Results Nimesulide effects a dose-dependent and time-dependent growth inhibition on QBC939 cells. High concentration of nimesulide (200 ?mol/L) not only inhibits the growth of QBC939 cells but also induces apoptosis cell nuclear condensation and apoptotic bodies were seen by transmission electron microscopy. Immunocytochemistry and RT-PCR shows upregulation of bax and down regulation of bcl-2 and survivin. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 in vitro by induction of apoptosis in a dose- and time- dependent manner.

Objective To establish a rapid assay for the determination of methotrexate (MTX) in serum.Method The assay was based on the ultraviolet absorbance of methotrexate at 306 nm. The separation of the drug was done by high performance capillary electrophoresis (HPCE). The bare fused-silica capillary was 60 cm in total length, 50.5 cm in efficient length and 75?m in diameter. The voltage of 25 kV was applied. The running buffer was 75 mmol/L phosphate, pH7.4. The performance of methodology was evaluated.Result The complete separation of MTX was achieved within 10 min. The linearity of the assay was from 1.1 ?mol/L to 1100.0 ?mol/L. The minimal detection limit was 0.55 ?mol/L.The recovery of MTX was from 88.2% to 98.2%. Within-run precision was 4.2% and between-run precision was ~5.4%. Conclusion The result indicated that the method was an effective method for clinical and scientific research with advantages of rapidity, simplicity and accuracy.

OBJECTIVE:To provide identification reference for the clinical use of Cinnabaris. METHODS:TCM micro-macro-scopical identification method and microscopic identification method were used. RESULTS:The micro-macroscopical characteristics were obtained:irregular granule or sheet block,different forms,wide bright red,opuque translucent with some luster; some sam-ples showed irregular lump with big shape,red scale on surface,dulling or gray-black. The were microscopic characteristics ob-tained:different forms of irregular granule,some sheet block,wide bright red,with some luster,occasionally with yellow gran-ules. CONCLUSIONS:The method for micro-macroscopical identification and microscopic identification of Cinnabaris is simple and convenient,and it can be used for the rapid verification of Cinnabaris.

Acute myeloid leukemia ( AML) is the most common acute leukemia in adults and has the highest death rate of all leukemias. Compared with other hematologic malignancies , there was only a small increment in the 5-year relative overall survival for patients with AML in the last 40 years, despite the advancement in our understanding of AML .CD123 is an AML-associated antigen that expresses at a high level in leukemic stem cells and leukemic blasts and a low level in normal hematopoiet-ic stem cells.As an attractive surface target for AML therapies , immune-based therapies targeting CD 123 are being developed re-cently, especially chimeric antigen receptor ( CAR) T-cell-based immunotherapy .Preclinical data have demonstrated that CD 123 CAR-T cells exhibit potent antileukemic activity and various im-pacts on normal hematopoiesis .This will probably be a promis-ing treatment for patients with relapsed/refractory AML.

Objective To explore the feasibility and significance of detecting Cyclin D1 and bcl-2 with flow cytometry in the diagnosis of B-cell lymphoma.Methods 45 patients patients with confirmed hematologic diseases in final diagnosis were collected and recorded 45 cases in detail included in the study.25 healthy persons showing normal results in routine physical examinations and laboratory tests were also selected and matched according to the age and gender.In addition,according to the age and gender matching principle,selected routine physical examination and all tests were within the normal range in 25 healthy persons as a healthy control group.Select the pathologically confirmed Cyclin D1 or bcl-2 positive cases were selected as the positive control group,select and the negative cases as the negative control group.Four-color monoclonal antibodies directly immunofluorescence monoclonal antibody labelinged with immunofluorescence were used to analyze the surface and cytoplasmic antigens,and multi-parameter flow cytometry was used to investigate the peripheral blood Cyclin D1 and bcl-2 subsets in lymphoma patients.Results The normal values of Cyclin D1 MFI and bcl-2 were obtained from the 25 normal volunteers by analyzing the (x)±s values and 95 ％ confidence interval,thus establishing the diagnostic criteria.Compared to the healthy control (0.356±0.159,1.938±0.324),all B-cell lymphoma patients had increased expression in Cyclin D1 (1.824±0.312)and bcl-2 MFI (4.259±0.541) in their peripheral blood,results showing statistical significance (P ＜ 0.01).Patients with different subtypes of lymphoma expressed differing Cyclin D1 MFI.In patients with Hodgkin' s lymphoma,Cyclin D1 MFI (0.386±0.112) was significantly lower than that in the non-Hodgkin' s lymphoma patients (1.623±1.987) (P ＜ 0.01).Further,in non-Hodgkin' s lymphoma patients,mantle cell lymphoma was 100 ％ positive for Cyclin D1,while the remaining subtypes were negative.Patients with different subtypes of lymphoma also showed differences in bcl-2 expression.In the Hodgkin' s lymphoma,bcl-2 (2.045±0.877) was significantly lower than the non-Hodgkin' s lymphoma (4.045±0.499) (P ＜ 0.01).In non-Hodgkin' s lymphoma,T cell lymphoma expressed the lowest,while MCL and FL showed 100 ％ positivity.In patients with positive Cyclin D1 or bcl-2,the mean fluorescence intensity level of Cyclin D1 (before treatment 3.099±0.349; after treatment 1.008±0.279) or bcl-2 (before treatment 7.814± 1.030,after treatment 3.131±0.522) was significantly lowered after treatment (P ＜ 0.01).Conclusion Using the method of flow cytometry for detecting Cyclin D1 and bcl-2 in lymphoma cells is feasible,and it can be applied clinically to evaluate the treatment.

Objective To explore the neutrophil to lymphocyte ratio (NLR) and its relationship with the effect of chemotherapy and prognosis in patients with diffuse large B-cell lymphoma (DLBCL).Methods The clinicopathological characteristics and outcome of 51 patients with DLBCL diagnosed by pathological biopsy and immunohistochemistry who received CHOP or R-CHOP regimen were collected and reviewed.According to the median of NLR,the patients were divided into low NLR group (NLR≤2.32) and high NLR group (NLR＞2.32).The prognostic influence of the NLR on overall survival (OS) was studied by Kaplan-Meier method and Log-rank test.To evaluate the independent prognostic relevance of NLR,univariate and multivariate Cox regression models were applied.Results The complete response (CR) rates of the low and high NLR groups were 71.4 ％ (20/28) and 39.1％ (9/23),respectively (P =0.02).The OS in the low NLR group was significantly better than that in the high NLR group (1,2 and 3-year OSs were 96.4 ％,90.4 ％ and 72.3 ％ vs 63.9 ％,52.7 ％ and 42.2 ％,respectively,P =0.009).Univariate and multivariate Cox regression models analysis showed that NLR ＞ 2.32 was an independent prognostic factor (P =0.016).Conclusion An elevated NLR before treatment indicates the poor effect of chemotherapy and prognosis of patients.NLR is an independent prognostic factor for DLBCL.

Objective To evaluate the diagnostic significance of detecting serum-DNA concentration and clonal gene rearrangement in lymphogenous malignant patients.Methods Serum-DNA in 72 diagnosed patients were measured and analyzed by SPSS11.0.Serum and mono-nuclear cells samples were collected. IgH CDRⅢ, TCR?V9 region were amplified by PCR.Results The serum-DNA concentration of disease groups were (418?172)?g/ml,(426?192)?g/ml, (388?170)?g/ml and(400?110)?g/ml, each was higher than those of control group(77?47)?g/ml(P0.05).Conclusion The concentration of serum-DNAwere higher in lymphogenous malignant patients and tumor associated DNA could be easily detected, so they have important diagnostic value in lymphogenous malignant patients.

OBJECTIVE To investigate the etiology of acute respiratory tract infection(ARI)in Hefei area and risk factors that may influence the distribution of common pathogens.METHODS Direct immunofluorescence assay was applied to detect the respiratory syncytial virus(RSV),adenovirus(ADV),influenza virus A(FluA),influenza virus B(FluB),parainfluenza virus PIV(1,2,3)and real time fluorescent quantitation polymerase chain reaction (FQ-PCR)was applied to measuring Mycoplasma pneumoniae(MP)and Chlamydia pneumoniae(CP)in nasopharyngeal secretions and sputum specimens.RESULTS Among 530 cases included in this study,421 cases (79.43%)showed positive viral etiology.ADV accounted for 73(13.77%),FluA-68(12.83%),FluB-56 (10.56%),RSV-48(9.05%),PIVl-47(8.86%),PIV3-42(7.92%),PIV2-33(6.22%),MP-32(6.03%)and CP-22(4.15%).The detected positive rate of pathogens isolated in winter was the highest(85.07%).CONCLUSIONS Acute upper respiratory tract infection(AURTI)is common and more than 75%pathogens in Hefei area are virus in which the most commonly virus is ADV.Meanwhile atypical pathogens of infection should not be ignored.

Objective:To improve the clinical recognition of hereditary fibrinogen deficiency.Methods:The diagnosis and treatment process of a patient with acute promyelocytic leukemia (APL) complicated with hereditary fibrinogen deficiency who was admitted to the second Affiliated Hospital of Anhui Medical University in December 2018 was retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was initially diagnosed as APL, and the complete remission was obtained after dual-induction therapy of all-trans retinoid acid and arsenous acid. During the first consolidation treatment, repeated reviews of fibrinogen fluctuated between 1.0-1.5g/L, and further improving the fibrinogen gene sequencing to diagnose APL combined with hereditary fibrinogen deficiency.Conclusion:For APL patients in remission who have decreased fibrinogen for many times and patients with hereditary fibrinogen deficiency who have significantly decreased fibrinogen in a short period, bone marrow biopsy and genetic testing should be further conducted to determine the pathogenesis.