Objective To investigate the efficacy of imatinib combining with allogeneic hematopioetic stem cell transplantation or chemotherapy for bcr-abl positive acute lymphoblastic leukemia (ALL). Methods 12 cases were diagnosed on morphology, cytochemistry, immunophenotype and bcr-abl fusion gene. The induction is imatinib (400 mg/d) combining chemotherapy. 8 cases accepted allogeneic hematopoietic stem cell transplantation after complete remission (CR). If bcr-abl became positive, the patient was treated with imatinib (400～600 mg/d). 3 cases were tested with imatinib alternating chemotherapy after cr. Results 11 patients gained CR, CR rate 91.7 %; 5 patients (41.7 %) became bcr-abl negative through 2 courses induction. 3 cases relapsed after transplantation. 2 cases relapsed in imatinib combining chemotherapy group. The median remission interval is 16 months (imatinib combining transplantation group) and 10 months (imatinib combining chemotherapy group) (P <0.01) respectively. The median survival time is 18 months (imatinib combining transplantation group), and the other group (imatinib combining chemotherapy) is 12 months (P <0.01). Conclusion Imatinib combining chemotherapy achieved high CR rate for the bcr-abl positive ALL. Imatinib combining allogeneic hematopoietic stem cell transplantation is superior to imatinib combining chemotherapy for CR patients.

Incidence of cancer is increasing year by year,diagnostic techniques are continually advanced,and the numbers of anti-cancer drugs are growing,but the efficacy has not been fundamentally improved.Surgery,chemotherapy and radiotherapy can only kill the part of mature tumor cells,but can not kill the cancer stem cells,and eventually leading to tumor recurrence.While acupuncture,qigong,psychology comprehensive treatment can inhibit tumor growth and reduce metastasis and recurrence,improve quality of life and prolong survival.We should update our recognition,to carry out the study on the treatment of tumor by TCM with large sample and multiple centers,to demonstrate its low toxicity,low costs and efficiency.

BACKGROUND:The proximal tibial fractures within joint are a common type of fracture, and clinical treatment is difficult. The implant internal fixation is a commonly used method of treatment. However, the choice of the optimal implant internal fixation is always the key problem in clinical research. OBJECTIVE:To explore the curative effect of different implant internal fixation for the treatment of intra-articular proximal tibial fractures and to obtain the better treatment through the comparative analysis. METHODS:A total of 32 patients of intra-articular proximal tibial fracture, who were treated with different implant internal fixation for the treatment of proximal tibia, were selected from the Department of Orthopedics, People’s Hospital of Zezhou County of Jincheng City and Second Affiliated Hospital of Shanxi Medical University from September 2010 to September 2013. These patients were divided into two groups according to fixation methods (n=16). Observation and control groups received LISS locking plate and common plate, respectively. The amount of bleeding, operation time and incision length were observed in both groups. Patients were fol owed up for 1 to 12 months after treatment so as to record fracture healing time, ful weight bearing time and excel ent and good rate and to observe the incidence of complications, including joint stiffness, loosening, nonunion and infection. RESULTS AND CONCLUSION:Incision length and operation time in the observation group were significantly shorter than that in the control group, and the amount of bleeding was significantly lower in the observation group than that of control group (al P<0.05). The healing of fracture and the ful weight bearing time were significantly shorter in the observation group than that in the control group (both P<0.05). The incidence of complications was significantly lower in the observation group (13%) than in the control group (46%;P<0.05). The total excel ent and good rate of repair was significantly higher in the observation group than in the control group (94%, 69%, P<0.05). These results confirm that compared with the traditional plate fixation, internal fixation with LISS locking plate for treating intra-articular proximal tibial fractures can get better therapeutic effects, greatly reduces the injuries to soft tissue and fracture blood supply, promotes fracture healing and restores joint stability.

Objective To observe the efficacy of imatinib on the treatment of bcr-abl positive essential thrombocythemia (ET). Methods A case of bcr-abl positive ET resistant to hydroxyurea (HU) treating with imatinib (200～400 mg/d) was reported and related literatures were reviewed. Results A case of bcr-abl positive ET was initially treated with 1.5～2.0 g/d HU, the platelet count decreased to 562x109/L after 4 weeks; however, the platelet count increased to (1020～1330)×109/L treating with same dose of HU 16 months later. With the elevation of HU to 3.0 g/d, platelet count was still high(1290～1780)x109/L companied with the very low white blood cell count(0.3～0.9)×109/L. While treating with imatinib (400 mg/d) for 1 month,the platelet count decreased to 390×109/L and white blood cell count was 0.5×109/L; Furthermore, treating with 200×300 mg/d of imatinib, the platelet and white blood cell count recovered in normal after 1 month,and bcr-abl fusion gene negative 2 months later. Conclusion Imatinib may be the effective targeting drug for the bcr-abl positive ET, and the bcr-abl positive ET is sensitive to low dose imatinib.

Linoleic acid (C18∶2n-6) and ?-linolenic acid (C18∶3n-3) are found widely in fungi, plants and some lower animals. However, they can not be synthesized in mammals due to lack of △12 and ?-3 fatty acid desaturases. To enable endogenous production of essential fatty acids in mammalian cells, here the stable expression of a Caenorhabditis elegans gene FAT-2 encoding △12 fatty acid desaturase in CHO cells was reported. First, the FAT-2 coding sequence was cloned by RT-PCR. To facilitate high level synthesis of heterogeneous protein, the codon usage of the fatty acid desaturase genes was optimized according to the codon preference of mouse by site-directed mutagenesis, 2 synonymous mutations were introduced into FAT-2 gene by overlapping PCR. The codon-modified gene was finally fused to pBudCE4.1 vector (Invitrogen) under the control of CMV promoter. The expression vector pBudCE-FAT2 was linearized with NheⅠ, and then transfected CHO cells, the cells were under Zeocin selection for nine days and then propagated, then the transfected cells were harvested. The genome and total RNA were isolated for PCR and Norhern blot ananlysis. The results revealed that FAT-2 gene has been integrated into the genome of CHO cells and expressed properly. Fatty acids of total cellular lipids were analyzed by gas chromatography. The results indicate that the expression and function of △-12 fatty acid desaturase resulted in accumulation of linoleic acid. The levels of linoleic acid in transgenic cells were 2.4-fold higher than those in wild-type cells. The moderate linoleic acid in CHO cells was derived from cell culture media uptaken by cell membrane. The results demonstrate that a heterogenous desaturase gene can function well in mammalian cells and prove that transgenic approach is an efficient strategy for changing fatty acid composition of mammals.

Objective To investigate sequence variations of 12 miRNA genes in multiple myeloma(MM) in order to find whether sequence variations in miRNA genes are associated with tumorigenesis and discuss the clinical significance of MM associated with miRNA genes mutations. Methods The miRNA gene mutations in 20 cases of MM, 4 MM-derived cell lines and 20 controls were detected by the methods of polymerase chain reaction single stranded conformation polymorphism (PCR-SSCP) and silver staining technique. Both clinical features and laboratory results were analyzed simultaneously. Results The electrophoretic patterns showed a total of three variations in miR-19a, miR-19b and miRNA-335,which were observed in 3 MM cells (15 %, 3/20). We also found variations of miRNA-335 in MM-derived cell lines KM-3and RPMI8226. However, no sequence alteration in the miRNA genes was observed in our set of controls. One of the three MM patients died, and two of them were detected mutations at the terminal stage of the disease.Conclusion A relative high frequency of miRNA gene mutation was found in MM and MM derived cell lines, which suggests possibility of a main mechanism underlying tumorigenesis. And, detecting miRNA gene mutations in MM might be benefit to evaluate the progression and prognosis of disease.

Objective To construct a recombinant adenovirus vector of Hairpin RNA specific for MRP1 gene and study its inhibition of MRP1 gene expression in K562/AS2 cell line resistant to AS_2O_3 (ATO). Methods A MRP1-specific hairpin RNA recombinant adenovirus vector was constructed and used to infected K562/AS2 cells. Expression level of MRP1 mRNA detected by real-time fluorescent quantitative PCR. MRP1 protein detected by flow cytometry. MTT method was used to detected the cytotoxicity of ATO and etoposide. Results MRP1 mRNA and protein expression level in K562/AS2 cells before and after the pAd-MRPl-shRNA adenovirus infection was (34.70±0.28 vs 4.19±0.03, P <0.05) and (26.40±0.16 vs 10.85±0.37, P<0.05), respectively. RR of K562/AS2 to arsenic trioxide and etoposide was (11.4078±0.3183 fold vs 1.6126±0.3015 fold, P<0.05) and (5.9141 ±0.0149 fold vs 1.7664±0.1038 fold, P <0.05), respectively. The reversal fold of ATO and etoposide was (7.2409±1.3668) and (3.3555±0.1886), respectively. Conclusion Successfully constructed pAd-EGFP-U6-shRNA-MRPl adenovirus vector, the vector of infection K562/SA2 cells can inhibit MRP1 gene expression and reverse the resistance of the ATO and etoposide.

Objective To investigate the reversal effect of Topo Ⅱα-shRNA and Topo Ⅱβ-shRNA on Topo Ⅱ gene in K562/AS2 cells. Methods Three pieces of Topo Ⅱα-shRNA and three pieces of Topo Ⅱβ-shRNA were designed,synthesized and transfected into K562/AS2 cells by liposome. Expression level of Topo Ⅱα and Topo Ⅱβ mRNA were determined by real time fluorescence quantitative PCR. The expressions of Topo Ⅱα and Topo Ⅱβ protein were assayed with flow cytometer. Results After treated with Topo Ⅱα-shRNA or Topo Ⅱβ-shRN A for 24 hours,the expression level of Topo Ⅱα mRNA and Topo Ⅱβ mRNA protein in K562/AS2 cells decreased at most (78.22±0.01) %,(31.17±1.27) % (P <0.05),and (57.36±0.01)%,(23.98±1.22) % (P <0.05) respectively. Conclusion The expression of Topo Ⅱ gene can be down-regulated after infected with Topo Ⅱ-shRNA in K562/ AS2 cell line.

ObjectiveTo investigate the gene expressions of TAZ and Wnt/β-catenin on the postosteogenic cells of mesenchymal stem cells(MSC)in multiple myeloma(MM)patients and to explore the potential therapeutic target of multiple myeloma bone disease (MBD).MethodsBone marrow mononuclear cells MNC from MM and controls were isolated,cultured,expanded and then induced to osteogenic differentiation.Realtime quantitative RT-PCR was employed to detect the osteogenic markers (TAZ,Wnt/β-catenin,OPN,OC,ALP and Cbf α1); and alizarin red staining for mineral deposition.The mRNA expressions of TAZ and Wnt/β-catenin in the two groups were analysed.ResultsAlizarin red staining was positive and the red calcium nodules were appeared on the post-osteogenic cells of MSC.The mRNA expressions of OC,ALP and Cbf α1 were 2.0958±0.5665,2.6670±0.3847,0.8463±0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly higher than those of pre-osteogenic cells(1.3487±0.9291,1.1452±0.6054,0.4439±0.2945) (t =2.171,6.709,2.795; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP and Cbf α1 were 2.1096±0.8267,2.8991±0.3531,4.3045±0.2844,1.3273±0.4075,respectively,on the post-osteogenic cells of MSC in the controls,which were significantly higher than those of pre-osteogenic cells (1.2200±0.9091,0.8780±0.3927,1.9161±0.2684,0.6736±0.2513) (t =2.289,12.103,25.134,4.411; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP,Cbf α1 were 1.2710±0.5636,2.0958±0.5665,2.6670± 0.3847,0.8463+0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly lower than those of control groups(2.1096 ±0.8267,2.8991 ±0.3531,4.3045±0.2844,1.3273±0.4075) (t =-2.650,-3.805,-10.822,-2.841; all P ＜ 0.05).The mRNA expression of TAZ and β-catenin were 2.2315±1.0723 and 0.5801±0.2159 on the post-osteogenic cells of MSC in MM patients,which were significantly lower than those of control groups (4.4140±0.8325,0.9516±0.2920) (t =±5.085,-3.235;both P ＜ 0.05).ConclusionThe gene expressions of OPN,OC,ALP and Cbf α1,the osteogenesis related genes,are increased in post-osteogenic cells of MSC,which showed the MSC have been successfully induced to osteoblasts.Comparing with control groups,the osteogenic potential of MSC in MM patients is lower.Based on the above research,TAZ and Wnt/β-catenin may present a novel target for the future therapy of MBD.

AIM:To explore a more accurate and reliable pathological model of the chronic bronchitis , which has improved from the former single-factor modeling method of the disease .METHODS:The mice in complex group were treated with lipopolysaccharide ( LPS) by tracheal injection on the 1st day and nasal drops on the 14th day, and from the 2nd day to 30th day, the animals were given passive smoking and sulfur dioxide ( SO2 ) inhalation ( except on the 14th day).The mice in SO2 group were exposed to SO2 2 min per day, while in smoking group, the mice were exposed to smoke for about 1 h per day (4 cigarettes each time until one pack of cigarettes were burning up ).In LPS group, the mice had tracheal injection of LPS on the 1st day and nasal drops of LPS on the 14th day and 30th day.Every modeling process las-ted for 30 days.After modeling, the improvement of chronic bronchitis model was evaluated by testing the general condi-tions of the mice , analyzing leukocyte count in bronchoalveolar lavage fluid ( BALF ) , and observing the morphological changes of the bronchial and lung tissues .RESULTS:After modeling, the mice in every model group experienced symp-toms including wet nose, cough, dry and lusterless hair, arched back and curled-up body, showing inactive, and slow down in response .The mice in complex group gained the lowest weight compared to other groups .From each model group , the inflammatory cells infiltrated evidently around the bronchial walls , especially in the bronchial cavity , and the mucilage secretion in the airway increased .The total number of leukocytes in BALF increased significantly in complex group .The in-flammatory cell count in the lung tissue indicated that the mice in complex group had significantly higher levels of inflamma -tory cell infiltration.Besides, the comparison between smoke group and LPS group was statistically significant .CONCLU-SION:Smoking, SO2 inhalation and LPS injection induce bronchial lung disease in mice , and the complex chronic bron-chitis mouse model is a better model with the pathological changes of bronchus , lung tissue and BALF , and pathogenesis of chronic bronchitis .