Objective To study the effect of fragile histidine triad (FHIT) gene on pancreatic adenocarcinoma cells growth and tumorigenicity and explore the mechanism of FHIT gene in suppressing the deve lopment of pancreatic adenocarcinoma. Methods By the method of liposome transfection, pRC/CMV FHIT plasmid was transfected into 1990 cell lines which lose all of FHIT gene. Integration and expression of exogenous FHIT gene were confirmed by RT PCR and Western blot technique. 1990 pFHIT cell growth was observed in regular culture medium and tumorigenicity in nude mice. Its DNA was analyzed by electrophoresis. Results The growth of the cells transfected with FHIT gene (named as 1990 pFHIT cells) was suppressed significantly, and the tumorigenicity of the 1990 pFHIT cells was dramatically inhibited in nude mice as compared with that of the parental 1990 cells. Significantly increased apoptosis in 1990 pFHIT was found. Conclusions The growth and tumorigenicity of pancreatic adenocarcinoma cell can be inhibited by transduced exogenous FHIT gene. It's spectulated that FHIT suppress the development of pancreatic adenocarcinoma by the path of apoptosis.

Objective To estimate whether the ligation of bilateral internal carotid artery in combination with one vertebral artery can lead to chronic cerebral hypoperfusion. Methods The sham operation, 2?VO and 3?VO rat models were subjected to the matching operation. Four weeks after operation,the cortical blood flow was determined. The learning and memory abilities were measured with Morris water maze test eight weeks later,then the rats were sacrificed to observe the morphological change of hippocampal CA1 region. Results Compared with the sham operation group((47±8.797)ml·min-1·100 g-1),the cerebral blood flow of 2?VO((24.30±8.999)ml ·min-1·100 g-1) and 3?VO((9.870±2.208)ml·min-1·100 g-1) were significantly decreased (P<0.01). Compared with the sham operation group((8.33±4.88)s),escape latencies of Morris water maze of 2?VO group ((14.78±7.84)s) and 3?VO group((14.86±7.96)s) in the fifth days also presented significantly increased (P<0.01),but rare difference between the two groups. Compared with the sham operation group[ (37.20±9.21) s, (10.01.±2.91)times],the target quadrant swimming time and crossing times of 2?VO group((20.13±5.80)s, (6.60±3.19)times) and 3?VO group((20.05±5.76)s,(6.55±2.59)times) in the fifth days also presented signifi?cantly decreased (P<0.01). There were distinct pathomorphology changes in hippocampal CA1 region of the two groups. Conclusion The ligation of bilateral internal carotid artery in combination with one vertebral artery can lead to chronic cerebral hypoperfusion,and can make the similar ethology representation with the 2?VO models.

10.3969/j.issn.1008-9691.2013.03.005

Objective To investigate the effect of the overexpression of NeuroD1 on mediating transdifferentiation of spinal reactive astrocytes into neurons. Methods Spinal cord astrocytes were cultured from the SD rat, and reactive astrocytes were prepared by scratches treatment. Cells were divided into blank groups (NV group), control virus group (GFP group) and NeuroD1 virus group (NeuroD1 group). At 7 d after scratches treatment, GFP and NeuroD1 groups were infected with retroviruses carrying the GFP gene and and GFP gene plus NeuroD1 gene, respectively,whereas NV group was not infected with the virus. Twenty-four hours late, the culture medium were replaced by neuron conditioned medi?um. Cell morphology was examined at 1, 2, 3, 5, 7 and 14 d. DCX positive and NeuN positive cells were detected at 7 d and 14 d after infection by using immunofluorescence staining method, respectively. Results After replacement with the neuron conditioned medium, the nucleus was obviously plump, the cytoplasm was thin and neurites was reduced and ex?tended. Compared with the NeuroD1 group, neurites of NV group and GFP group were shorter with many branches and the nucleus was smaller. At 7 d after infection, cell morphology of NV group and GFP group gradually recovered, but cell morphology of NeuroD1 group did not. Compared with NV group and GFP group, NeuroD1 group had more DCX(9.84 ± 2.06%)and NeuN(8.25±2.78%)positive cells [F values 40.107 for DCX and 21.73 for NeuN (P<0.05)]. Conclusion The overexpression of NeuroD1 can mediate the transdifferentiation of spinal reactive astrocytes into neurons.

Objective To investigate the effect of mild hypothermia on proliferation and differentiation of neural stem cells (NGCs) in hippocampal subgranular zone after traumatic brain injury (TBI) and the underlying mechanism.Methods SD rats were divided into sham-injured group (only left dura mater exposed),hypothermia group (sham injury + mild hypothermia therapy for 72 hours),TBI group (unilateral fluid percussion was used to generate severe TBI),and TBI + hypothermia group (TBI + mild hypothermia therapy for 72 hours) according to the random number table,with 8 rats per group.Hippocampal homogenates or brain tissues were harvested after BrdU (100 mg/kg) was intraperitoneally administered to rats once a day for 7 days postTBI.Expressions of BrdU and double cortin in hippocampal subgranular zone were respectively detected by immunohistochemical or immunofiuorescence staining.Level of Sirt1 (silence information regulatory proteins,Sirt1) in hippocampus was detected by Western blot.Results BrdU-and double cortin-positive cells in rat hippocampal subgranular zone greatly increased at 7 days after TBI in comparison with sham-injured group (P ＜ 0.01).Moreover,BrdU and double cortin in rat hippocampal subgranular zone in TBI + hypothermia group was significantly higherthan that in TBI group [(257.4 ± 34.3) vs (196.4 ± 23.8) ; (346.4 ± 42.2) vs (245.7 ± 33.2),P ＜0.01].Moreover,mild hypothermia reversed TBI-induced over-expression of Sirt1 [(0.62 ± 0.075) vs(1.18 ± 0.11),P ＜ 0.01].Conclusion Mild hypothermia therapy can promote proliferation andneuronal differentiation of NSCs in hippocampal subgranular zone after TBI and the possible mechanismmay relate to the inhibition of over-expression of Sia1.

Objective To investigate the effect of silence regulatory protein 1 (Sirt1) on axonal outgrowth. Methods The hippocampal neurons was first isolated in vitro from rat embryos. The distribution and expression of Sirt1 were then detected 72 h later. The down-regulation of Sirt1 was induced by RNAi technology and up-regulation of Sirt1 was in-duced by overexpression of Sirt1 and resveratrol (RES). Immunofluorescence staining was used to examine the axon length. Results Immunofluorescence staining showed that Sirt 1 was located in neuronal cell body and neurite, especially in the distal axons. Down-regulation of Sirt1 significantly decreased axonal length compared with siRNA control group [(178.3 ± 3.2) μm vs. (110.2 ± 18.30) μm, P< 0.01 ]; Overexpression of Sirt1 significantly increased axonal length com-pared with eGFP control group [(178.3±3.2)μm vs (310.6±39.5)μm, P<0.01 ];Activation of Sirt1 by RES treatment al-so significantly increased axonal length compared with vehicle control group (DMSO treated group) [(291.7±13.2)μm vs. (525.1±49.1)μm, P<0.01 ]. Conclusions Sirt1 plays a key role in axonal growth which may be used as a potential thera-peutic target of axon regeneration.

Objective To investigate the relationship between serum cholesterol levels and immunoglobin types and clinical stages in the patients with multiple myeloma (MM). Methods We retrospectively analyzed the blood lipid levels in 65 patients with MM at diagnosis, including total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein Al (apo-Al) and apolipoprotein B (apo-B), and explored relationship between lipid parameters and immunoglobulin types or clinical stages in patients with MM. Thirty healthy persons were served as controls. Results Of the 65 MM patients, 53.85% were IgG type, 63.1 % were at stage Ⅲ. The levels of TC, HDL-C, LDL-C, apo Al and apo B in the patients with MM were significantly lower than that in the controls (P <0.05), and TG in MM patients was no difference with that in the controls (P >0.05). Except one case of IgD type, the levels of TC, HDL-C, LDL-C, apo Al and apo B in Ig G and Ig A types of patients were significantly lower than that in the light chain type among other 64 cases (P <0.05), and TG levels in different immunoglobulin types was found no statistical differences. The levels of TC, HDL-C, LDL-C and apo A1 in the patients with stage Ⅲ were lower than that of stage I and controls (P <0.05), furthermore, the level of LDL in stage Ⅱwas lower than that in stage Ⅰ. Conclusion Hypocholesterolemia are seen in the patients with MM and serum cholesterol levels are related to MM staging.

Objective:To observe the dynamic variation of serum ferritin (SF), folic acid, and vitamin B_(12) levels in patients with acute promyelocytic leukemia (APL) at different disease stages. Methods:Serum SF, folic acid and vitamin B_(12) levels were successively tested in thirty-six patients with primary APL every 1 to 3 months by using chemiluminescence analysis. Five different disease stages were selected as dynamic observation time points: first diagnosed, first complete remission (CR1), six months after CR1, relapsed stage,and CR1 for three years. Results:There were 75.0%(27/36)of patients with abnormal high levels of SF, 77.8% (28/36)of patients with abnormal low levels of folic acid, and 100%(36/36)of patients with increased vitamin B_(12) levels in first diagnosed stage. The number of patients with abnormal variations of SF, folic acid and vitamin B_(12) level was decreased in CR1 stage compared with those in first diagnosed stage (SF: P<0.05;folic acid and vitamin B_(12): P<0.01). The serum SF, folic acid and vitamin B_(12) levels tended to recover step by step with chemotherapy. At six months after CR1 the three parameters of most patients recovered to normal levels. APL was relapsed in 4 patients after 1-year CR. Both SF and vitamin B_(12) levels were increased and the folic acid level was decreased compared with those before replase, but the difference had no significance (P>0.05). The serum SF, folic acid and vitamin B_(12) levels were in normal ranges in the patients who had 3-year CR. Conclusion:The serum SF, folic acid and vitamin B_(12) levels had dynamic variation in APL course. Increase in serum SF and vitamin B_(12) as well as decrease in folic acid are related with the active degree of APL and its tumor load.

Objective: To evaluate the efficacy and prognostic factors in core binding factor (CBF) acute myeloid leukemia (AML) under current therapy modalities, therefore optimizing the treatment strategies. Methods: Standard cytological and immune methods including next generation sequencing (NGS) were used for risk stratification. Complete remission (CR) rate, disease-free survival (DFS) and overall survival (OS) were assessed by multivariate Logistic and Cox regression models in a total of 206 adults (aged 16-65 years) with CBF-AML, including 152 AML patients with t(8;21) and 54 with inv(16). Results: The CR rate of inv(16) patients after first course was 54/54(100%), significantly higher than that of t(8;21) patients [127/147(86.4%), P=0.005]. The fusion transcript level and KIT mutation were independent factors related to CR rate in t(8;21) patients (P=0.044 and 0.027; respectively). DFS and OS in inv(16) patients tended to be more superior than that in t(8;21) patients (P=0.066 for DFS; P=0.306 for OS; respectively). Multivariate Cox identified negative expression of CD₁₉ and female gender the independent predictors of inferior DFS in t(8;21) patients (P=0.000 for CD₁₉; P=0.006 for sex; respectively). Analysis of combining CD₁₉ with gender indicated that females/CD₁^9-subpopulation had significantly poor DFS than did males/CD₁₉⁺ ones (Bonferroni-P<0.000 01). The number of mutations in each patient, FLT3-ITD and additional karyotype abnormalities did not affect CR rate and DFS (all P>0.05). Conclusions Patients with inv(16) have better induction response than those with t(8;21). High level of fusion transcripts and positive KIT mutation are associated with low CR rate in t(8;21) patients. Negative CD₁₉ expression and female gender are independent predictors of inferior DFS in t(8;21) patients.