Objective To evaluate the enhancement of chemosensitivity of celecoxib, a specific cyclooxygenase-2 (COX-2) inhibitor, on leukemia HL-60 cell line in vitro, and explore the possible mechanisms. Methods MTT assay was used to assess the cytostatic efficacy of doxorubicin in the absent or present of different doses of celecoxib on HL-60 cell. The apoptosis of HL-60 cells was measured by flow cytometry (FCM). Gene expressions of Survivin was examined by reverse transcription-polymerase chain reaction (RT-PCR). Protein of survivin was detected by Western blotting. Results Celecoxib could increase the cytostatic efficacy of doxorubicin on HL-60 cells. HL-60 cells were treated with increasing doses of doxorubicin in absence or presence of celecoxib (5 μmol/L, 10 μmol/L), IC50 were 0.48 μg/ml, 0.25 μg/ml and 0.16 μg/ml, respectively. Doxorubicin combined with low dose of celecoxib could induce the down-regulation of mRNA and protein of Survivin. Apoptosis rate of HL-60 cells treated with both 0.10 μg/ml doxorubicin and celecoxib(5 μmol/L, 10 μmol/L) were (13.07±1.66) % and (22.36±1.84) %, respectively, while it was (5.72±1.25) % in HL-60 cells treated with 0.10 μg/ml doxorubicin alone, with significant difference (P＜0.01).Conclusion Celecoxib could enhance the chemosensitivity of doxorubicin on leukemia HL-60 cell, which involves in increasing the apoptosis of HL-60 cells by down-regulation expression of Survivin.

Objective To evaluate clinical treatment method and efficacy of newly diagnosed acute promyelocytic leukemia (APL),and analyze the relevant factors about the long-term survival.Methods The clinical data of 48 patients with newly diagnosed APL were analyzed retrospectively.All of them used alltrans retinoic acid (ATRA) combined with anthracycline as induction remission therapy.After induction remission,ATRA combined with chemotherapy was used as consolidation therapy,and ATRA,arsenic trioxide and conventional chemotherapy alternated as maintenance therapy.Short-term efficacy was analyzed.Patients were followed up,and the rates of overall survival (OS) and disease-free survival (DFS) were analyzed.Long-term efficacy was analyzed by COX proportional hazards regression models univariate analysis.Results The complete remission (CR) rate was 87.5％(42/48) in all 48 patients with APL.The time from treatment beginning to CR was (30.7 ± 4.6) d.Age was the only factor affecting the rate of CR.The rates of 1-year,3-year and 5-year OS were (87.5 ± 4.8)％,(85.4 ± 5.1)％ and (78.3 ± 6.7)％ in 48 patients with APL.The rates of 1-year,3-year and 5-year DFS were (97.6 ±2.4)％,(93.9 ±4.2)％ and (89.5 ± 5.9)％ in 42 patients with CR.COX proportional hazards regression model univariate analysis result showed that the patient' s age,gender,lactate dehydrogenase,diffuse intravascular clotting,risk stratification and bone marrow abnormalities promyelocyte ratio had no correlation with the rate of DFS (P ＞0.05).Conclusions ATRA combined with anthracycline as induction remission therapy,after induction remission ATRA combined with chemotherapy as consolidation therapy,and ATRA,arsenic trioxide and conventional chemotherapy alternated as maintenance therapy can get a higher rate of CR and long-term survival in patients with newly diagnosed APL.It is worthy of clinical application.

A novel method for analysis of three active components curcumin, demethoxycurcumin and bisdemethoxycurcumin in Curcuma longa L. was developed by HPLC coupled with electrochemical detection. Three curcuminoids were well separated on a C18 column and detected with high sensitivity. A mobile phase containing acetonitrile and 10 mM Na2HPO4-H3PO4 (pH 5.0) (50:50, v/v) was used. Good linearity was obtained in the range of 0.208-41.6, 0.197-39.4, and 0.227-114μM for curcumin, demethoxycurcumin and bisdemethoxycurcumin respectively. The limit of detection reached up to 10 ? 8 M, which was lower than that by UV detection. The relative standard deviations (RSDs) ranged from 1.06%to 1.88%for intra-day precision and from 4.30%to 5.79%for inter-day precision, respectively. The proposed method has been applied in real herb sample and recoveries ranging from 86.3%to 111%were obtained.

Background Intracorneal macrophages play a critical role in corneal neovascularization (CNV)by secreting relative chemokines.But macrophages are characteristic by heterogeneity which has different biologic functions under different induction or stimulation from microenvironment.Objective This study was to detect the effects of chemokine (C-X3-C motif) ligand 1 (CX3CL1) and chemokine (C-C motif) ligand 2 (CCL2) on macrophages in vitro.Methods CNV was induced by corneal alkali burn in the left eyes of 20 male BALB/c mice aged 7-8 weeks.The CNV was evaluated under the slit lamp microscope 4 days after alkali burn,and then the corneal sections were prepared after mice were sacrificed.The expressions of CCR2 and CX3CR1 in the corneal specimens were detected by histo-fluorescence staining.Human peripheral blood mononuclear cells were separated using density gradient centrifugation and incubated in RPMI-1640 medium containing 10％ fetal bovine seruml(FBS) with 30 μg/L granulocyte-macrophage colony-stimulating factor (GM-CSF).The cells were divided into CD68 +CCR2 group and CD68+CX3CR1 group,and the percentage of the CX3CR1 and CCR2 expressions in the infiltrated macrophages of corneal specimens and human monocyte-derived macrophages was assayed by flow cytometry.The cultured cells were stimulated using human recombinant CX3CL1 and CCL2 proteins,and real-time PCR was used to detect the relative expressions of angiogenesis-related factors in macrophages.Results CNV was found in corneas 4 days after alkali burn and the CNV onsets from corneal limbus to central zone observed by a slit lamp.CCR2 and CX3CR1 were expressed in the F4/80-positive macrophages in alikali burned corneas.The macrophages grew for two weeks and appeared more dead cells in without GM-CSF group,but in GM-CSF induced group,the number of macrophages was increased.The percentage of CX3CR1-positive cells was 75％ and that of CCR2-positive cells was 45％.Real-time PCR showed that expression level of vascular endothelial growth factor (VEGF) mRNA increased and that ADAMTS-1 mRNA or TSP-1 mRNA decreased on macrophages after CCL2 stimulation,with significant differences in the 150 mg/L CCL2 group compared with the control group (t =-5.09,P =0.03 ; t =3.01,P =0.04 ; t =4.27,P =0.02).However,the VEGF mRNA expression decreased and ADAMTS-1 mRNA and TSP-1 mRNA increased after CX3CL1 stimulation,showing significant differences between the 150 mg/L CX3CL1 group and the control group (t=6.35,P=0.O2;t=-2.92,P=0.04; t=-3.81,P=0.03).Conclusions These results suggest that the macrophages have high heterogeneity.CCL2-and CX3CL1-expressing macrophages can regulate the expressions of angiogenesis-related factors.Macrophage chemokine signal may be a good target for treatment of neovascular ocular disease.

Background and purpose:Differentiation of tumor tissue is an important factor on determining the prognosis of gastric cancer. This study aimed to investigate the expression levels and clinical signiifcance of gender determining region Y-box 2 (SOX2) gene and octamer binding factor 4 (OCT4) gene in gastric cancer tissues varying different differentiation degrees. Methods: Sixty cases with gastric cancer were recruited in this study. The gastric cancer tissues and corresponding normal mucosa of the 60 cases were obtained. The mRNA and protein level of SOX2, OCT4 gene are evaluated by the quantitative real-time PCR (qRT-PCR), Western blot and immunohistochemistry, respectively. The relationship between the expression levels of SOX2, OCT4 gene and clinical pathological parameters were also analyzed in this study. Results:The expression of SOX2 in both mRNA and protein levels had no signiifcant difference between the well-differentiated gastric cancer tissues and normal gastric mucosa (mRNA levels:t=0.1033, P>0.05;protein levels:t=0.116, P>0.05). However, both the mRNA and protein expression of SOX2 in patients with well-differentiated gastric cancer tissues were signiifcant higher than not only in patients with moderately differentiated gastric carcinoma (mRNA levels: t=12.48, P<0.05; protein levels: t=22.78, P<0.05) but also in patients with than poorly differentiated gastric carcinoma (mRNA levels:t=17.56, P<0.05;protein levels:t=30.00, P<0.05). In contrast to SOX2, both the mRNA and protein expression of OCT4 in patients with well-differentiated gastric cancer tissues were signiifcant lower than not only in patients with moderately differentiated gastric carcinoma (mRNA levels:t=13.23, P<0.05; protein levels: t=25.56, P<0.05) but also in patients with poorly differentiated gastric carcinoma (mRNA levels: t=12.10, P<0.05; protein levels: t=69.48, P<0.05). There was no significance of OCT4 mRNA and protein expression between the well-differentiated gastric cancer tissues and normal gastric mucosa (mRNA levels:t=2.436, P>0.05;protein levels:t=1.064, P>0.05). Immunohistochemical study demonstrated that the positive rate of SOX2 in patients with well-differentiated gastric cancer tissues (10/21) were higher than in patients with not only moderately differentiated gastric carcinoma (7/20) but also poorly differentiated gastric carcinoma (2/19, P<0.05), while the positive rate of OCT4 in cases with well-differentiated gastric cancer tissues (2/21) were lower than in cases with not only moderately differentiated gastric carcinoma (6/20) but also the poorly differentiated gastric carcinoma (12/19, P<0.05). There was no correlation between the expression of SOX2, OCT4 in gastric cancer and gender or age (P>0.05). Nevertheless, the expression of SOX2, OCT4 were positive or negative correlated with the pathological staging, the degree of inifltration and lymph node metastasis (P<0.05). Conclusion:Decreased SOX2 expression and increased expression level of OCT4 can promote the formation, development and invasion of gastric cancer and they may become biomarkers or the diagnosis, treatment and prognosis evaluation in gastric carcinoma.

Objective To search for the molecular figngerprint of ascaris lumbricoides in intrahepatic stones. Methods In 56 cases deoxyribonucleic acid (DNA) was exstracted from intrahepatic stones and surgically obtained bile duct tissues, stool and sercum examinations were performed. Based on the sequence and designed primers of ITS-2 of ascaris lumbricoides, polymerase chain reaction ( PCR) was performed. Results There were 12 positive results in 56 (21.4% ) cases of intrahepatic stones. Positive reaction appeared on the examinations of the stool on ascaris lumbricoides and the blood serum among 12 cases. Conclusions Ascaris lumbricoides DNA was found in human of intrahepatic stones, indicating that ascaris lumbricoides infection may play a role in the formation of intrahepatic stones.

Objective To investigate the ultrasonic imaging features of thyroid follicular adenoma for conducting the correct diagnosis and differentiation diagnosis.Methods The clinical and imaging data in 64 cases of pathologically proven thyroid follicular adenoma were analyzed on the maximal diameter of tumor,nodularity number,high and low echogenicity,peripheral halo,echo hom-ogeneity,calcifications,and so on.The misdiagnosis causes were investigated.Results The mass was mainly solid or cystic-solid mixed echo.The ultrasonic imaging features of thyroid follicular adenoma were non-peripheral halo or thin wall halo,hyperecho or isoecho,internal macrocalcifications and peripheral calcifications,homogeneous echo structure.Conclusion The ultrasonographic examination can provide the better diagnosis and differentiation diagnosis on thyroid follicular carcinoma.

Objective To study the MR images of patients with Alzheimer's disease (AD) for analysis of the volume changes ofbrainstem and deep brain nuclei,hoping to provide evidence for early clinical diagnosis.Methods Selected for this study were MRI images from the ADNI database from July 2006 to November 2010 of 31 AD patients (AD group),34 patients with mild cognitive impairment (MCI group),and 34 normal aged people (NC group).The follow-up MRI data of all the above subjects 2 years later were also collected.Software Freesufer was used to calculate and compare the volume changes ofbrainstem and deep brain nuclei in all the subjects.Results An identical trend was found concerning the MRI-based volumes of brainstem,hippocampus and deep brain nuclei (caudal nucleus and putamen) before and after follow-up between the 3 groups:AD group＜MCI group＜NC group,with statistically significant differences between groups (P＜0.05).Follow-ups revealed significant decreases in all the indicators in AD and MCI groups (P＜0.05),but no such differences in NC group (P＞0.05).Pearson relative analysis showed a positive correlation between the brainstem volume and the volume of deep brain nuclei (caudal nucleus and putamen) in AD patients (r=0.653,P=0.021;r=0.596,P=0.014).Conclusions As AD progresses,the brainstem and deep brain nuclei in AD patients continue to shrink in volume and there may be a positive correlation between them.Therefore,detection of the volume changes of the brainstem and deep brain nuclei contributes to the early diagnosis of AD.

Objective To investigate the morphological changes of the brainstem in patients with Alzheimer's disease (AD) and their relationship with hippocampal morphological changes.Methods Sixty AD patients (AD group) and sixty age-and gender-matched normal elderly (normal control group) were selected from the Alzheimer's Disease Neuroimaging Initiative (ADNI) database.The hippocampus and the brainstem of each subject were segmented and their normalized volumes were calculated.According to the hippocampal volume standard value (Z-score),AD patients were divided into two subgroups (hippocampal atrophy group (n=51) and hippocampal spared group (n=9)).A voxel-based morphology (VBM) study was also performed to investigate the morphological differences of the brainstem between the normal control group and the AD group,as well as between the AD subgroups.Results Compared with the normal control group,the brainstem volume in the AD group decreased significantly (16 741.31±1 739.11 vs 15 609.67±1 451.60,t=3.870,P=0.001).In AD subgroups,the volume of the brainstem in the hippocampal atrophy group was significantly smaller than that in the hippocampal spared group (16 556.30 ± 1 514.86 vs 15 442.62 ± 1 389.05,t=2.189,P=0.033).Pearson correlation analysis showed that Mini-Mental State Examination scores were positively correlated with the hippocampal and the brainstem volumes (r=0.590,P＜0.01;r=0.234,P＜0.05),and there was a positive correlation between the hippocampal and the brainstem volume changes in patients with AD (r=0.315,P=0.014).VBM results showed that both the bilateral midbrain and the bilateral pons in the AD group had significant atrophy compared with the normal control group (P＜0.05).In the AD subgroups,the bilateral midbrain and the left pons in the hippocampal atrophy group were significantly atrophied compared with the hippocampal spared group (P＜0.05).Conclusion The brainstem showed morphological changes in patients with AD,and the morphological changes of the brainstem in AD patients with different degrees of hippocampal atrophy were different,indicating the morphological changes of the hippocampus and the brainstem may have an interrelated relationship.

Objective To investigate the accuracy of sequential organ failure assessment (SOFA) scoring in emergency physicians in Beijing. Methods Emergency physicians from 8 hospitals in Beijing in January 2018 were demanded to complete a SOFA questionnaire which was developed on ''wenjuanxing'' website and submit via cell phone. All participants were divided into urban center group (UC group) and no-urban center group (NUC group) based on the hospital's location. The accuracy rate of components and total score of SOFA along with the mistakes were evaluated, and the results of the two groups were compared. Results ① The questionnaire was sent to 217 emergency physicians of the 8 hospitals, and 197 qualified questionnaires were received with 109 of NUC group and 88 of UC group, respectively, the total response rate was 90.8%. Compared with those from NUC group, UC physicians had older ages [years:37 (32, 42) vs. 34 (29, 40), Z = -2.554, P = 0.011] and higher education level [postgraduate degree 76.1% (67/88) vs. 40.4% (44/109), χ2= 25.327, P < 0.001], and more of them experienced SOFA scoring [62.5% (55/88) vs. 45.9% (50/109), χ2= 5.409, P = 0.020]. Other baseline characteristics such as gender, working years, professional title and training experience were not different between the two groups. ② The accuracy rate of total SOFA score was 62.4% (123/197) in the whole cohort, and UC group was lower than that of NUC group, but the difference was not significant [56.8% (50/88) vs. 67.0% (73/109), χ2= 2.141, P = 0.143]. While comparing the accuracy of individual variable/system of SOFA, the accuracy rate of norepinephrine of UC group was much higher than NUC group [80.7% (71/88) vs. 66.1% (72/109), χ2= 5.235, P = 0.022], but the accuracy of Glasgow coma scale (GCS) was much lower in NUC group [38.6% (27/70) vs. 81.6% (71/87), χ2= 30.629, P < 0.001]. Other variables of SOFA were not different between the two groups. ③Based upon the results of all submitted questionnaires, 566 mistakes were identified. It was indicated that the mistakes per capital was 2.9 in the whole cohort and in the two groups. The first type mistakes which caused by carelessness (including calculating error, filling error, choosing error) were 233 times. The calculating error in norepinephrine from NUC physicians was higher than the UC group [33.9% (37/109) vs. 19.3% (17/88), χ2= 5.235, P =0.022], there was no significant difference in any other first type mistakes between the two groups. The total second type mistakes caused by misunderstanding of SOFA (including using wrong variables, not using the worst value within 24 hours, and incorrect GCS score) were 333 times in the whole cohort. GCS error [61.8% (42/88) vs. 16.9% (14/109), χ2=32.292, P<0.001], and using urine output per hour instead of urine output per 24 hours [15.9% (14/88) vs. 4.6% (5/109), χ2= 7.162, P = 0.007] were much higher in UC group than NUC group. Conclusions The total accuracy of SOFA scoring in the investigated emergency physicians of 8 hospitals in Beijing was not good. Mistakes causing by carelessness or misunderstanding of score rules were similar. It is necessary to apply strict training in SOFA scoring.