[Construction and Identification of Acute Myeloid Leukemia NOD/SCID Mouse Model by Tail Vein Injection of THP-1 Cells].

OBJECTIVE: To construct NOD/SCID mouse leukemia model by using THP-1 cells. METHODS: Eighteen female NOD/SCID mice aged 3 to 4 weeks were randomly divided into control group, model group A and model group B (6 in each group). Before inoculation, each mouse was intraperitoneally injected with cyclophosphamide 2 mg/(kg·d) for 2 d, and the mice in model groups were inoculated with cells within 24 h after pretreatment. The mice in model group were inoculated with THP-1 cell suspension in logarithmic growth phase by 1×107 cells/group (group A) and 5×106 cells/group (group B), the mice in the control group were injected with the same amount of normal saline in the tail vein. The general situation was observed, blood routine test and peripheral blood leukocyte classification were performed at 7, 14, 21, 28 d of inoculation before the pre-treatment, and at the time sacrifice. Before dying, tissue of mice were collected and histological examination was performed. RESULTS: Pilereation, droopiness and hypkinesia could be observed from d 7 and d 10 of inoculation cells in model group. Compared with the control group, the body weight of the mice in model group A and B decreased significantly after 21 days of inoculation (P<0.01), and the white blood cell counts increased significantly after 28 days of modeling (P<0.01). Among them, the above-mentioned presentation in inoculation of 1×107 group A was the most significant. Histopathological sections showed diffuse infiltration of leukemia cells in the spleen of the model group. The immunohistochemistry results indicated that the leukemia cells were positive for anti-human CD13, which confirmed the successful establishment of the model. CONCLUSION: After pretreatment with intraperitoneal injection of CTX in NOD/SCID mice, the injection of 1×107 or 5×106 THP-1 cells in tail vein of each mouse can successfully construct an acute myeloid leukemia animal model. The tumor formation is more much faster by injection of high concentration THP-1 cells.

Epstein-Barr virus-encoded LMP1 regulated Pim1 kinase expression promotes nasopharyngeal carcinoma cells proliferation.

BACKGROUND: Epstein-Barr virus-encoded LMP1 plays a critical role in the carcinogenesis of nasopharyngeal carcinoma (NPC), but the mechanism remains elusive. We aimed to analyze the expression and clinical pathological significance of provirus integration site for Moloney murine leukemia virus 1 (Pim1) in clinical NPC, and to elucidate the effect of LMP1 on Pim1 expression and its mechanism. METHODS: Immunohistochemical staining was used to detect the expression of Pim1 in clinical NPC tissues and control nasopharyngeal chronic inflammation (NPI) tissues, and the correlation between Pim1 and clinical parameters of NPC patients was analyzed. The LMP1 stable expression cell line CNE1-LMP1-OV was constructed through infecting the well-differentiated nasopharyngeal carcinoma cells CNE1 with LMP1 overexpressing lentivirus. Then the in vivo experiments were conducted. RESULTS: Among 89 NPC patients, 48 cases (53.93%) were positive for Pim1, while only one case was Pim1 positive in 15 NPI controls (6.67%). Pim1 expression was not correlated with gender, age, smoking status and clinical classification of NPC patients, but positively correlated with T, N and M classification. CNE1-LMP1-OV cell line was successfully established, which displayed a higher cell proliferation ability and Pim1 expression. NF-κB inhibitor PDTC, PKC inhibitor GF109203X and STAT3 inhibitor Stattic significantly attenuated LMP1-induced Pim1 expression, and while AP-1 inhibitor SR11302 showed no inhibitory effect. Interestingly, Pim1 inhibitor quercetagetin significantly inhibited the proliferation of CNE1-LMP1-OV cells. CONCLUSION: LMP1 mediates Pim1 expression through NF-κB, PKC and STAT3 signaling, which promotes the proliferation of NPC cells and participate in the clinical progression of NPC.

Modified sequential therapy vs quadruple therapy as initial therapy in patients with Helicobacter infection.

AIM: To evaluate the efficacy and safety of modified sequential therapy and to compare modified sequential therapy with standard quadruple therapy for Helicobacter pylori (H. pylori) eradication. METHODS: In total, 200 consecutive patients who were diagnosed with H. pylori-infected chronic gastritis by electronic endoscopy and rapid urease testing from December 2012 to October 2013 were enrolled in this study. The patients had not previously received H. pylori eradication treatment, and were randomized into two groups. The patients in Group A (n = 101) were treated with ilaprazole + bismuth potassium citrate + amoxicillin and clavulanate potassium + levofloxacin, and the patients in Group B (n = 99) were administered a modified sequential therapy composed of ilaprazole at 5 mg bid and amoxicillin and clavulanate potassium at 914 mg for the first five days followed by ilaprazole at 5 mg bid, furazolidone at 100 mg bid and levofloxacin at 500 mg qid for the next five days. Four to six weeks after the end of treatment, a 14C-urea breath test was performed for all the subjects to confirm the eradication of H. pylori. The intention-to-treat and per-protocol eradication rates were determined. RESULTS: A total of 190 of the 200 patients completed the study. All 200 patients were included in the intention-to-treat analysis, whereas 190 patients were included in the per-protocol analysis. In the intention-to-treat analysis, the rates of H. pylori eradication in Groups A and B were 85.15% (86/101) and 81.82% (81/99), respectively. In the per-protocol analysis, the H. pylori eradication rates in Groups A and B were 88.66% (86/97) and 87.09% (81/93), respectively. No significant difference was observed (χ(2) = 0.109, P = 0.741) in the eradication rate between Groups A and B. The rates of adverse effects observed in the groups were similar at 6.19% (6/97) for Group A and 7.53% (7/93) for Group B (P > 0.05). No mortality or major morbidities were observed in any of the patients. Symptomatic improvements in the presentation of stomachache, acid regurgitation, and burning sensation were not significantly different between the two groups. CONCLUSION: Ilaprazole-based 10-d standard quadruple therapy does not offer an incremental benefit over modified sequential therapy for the treatment of H. pylori infection, as both treatment regimens appear to be effective, safe, and well-tolerated as initial treatment options.

[Observation on gametophyte development of Cibotium barometz].

OBJECTIVE: To observe the spores germinating process of Cibotium barometz, and understand the growth principle provided for experience for indoor culturing and further research. METHOD: The spores of C. barometz were cultured both in inorganic medium and in the soil from original habitat, and the whole process of spores germination and the development of gametophytic were observed under microscope. RESULT: The spores germinated about 1-2 weeks after being sowed, and the type of germination belonged to Vittaria-type. The prothallial plates formed in 25 days after being sowed, while hairs developed after the formation of the prothallial plate. The gametophyte formed about 40 days after being sowed. But the type of mature prothalli was cordate. The antheridia formed in 60 days after inoculation, while the archegonia developed in 10 days after the formation of antheridia. CONCLUSION: Soil based indoor culturing of C. barometz spores is practical and can be used for cultivation of C. barometz.