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 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.

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.

Enhanced green fluorescent protein( EGFP), myc epitope and polyhistidine metal-binding tag are often used as a marker for recombinant fusion protein in many gene expression vectors, each marker has its own function, EGFP emits green fluorescence for direct detection, myc epitope facilitates recombinant fusion protein detection using its antibodies, polyhistidine tag allows purification of recombinant fusion protein using resin.Hitherto, no a plasmid vector can integrate all of these functions. In this study we constructed a novel eukaryotic expressive plasmid, designated as pcDNA6/myc-his-EGFP B, which integrated the functions of EGFP, myc epitope and polyhistidine tag. Importantly, a linker octo - peptide in N terminal of EGFP was designed using LINKER program. A DNA fragment encoding a putative protein containing a signal peptide of human interleukin 2(IL-2) in N terminal was cloned into pcDNA6/myc-his-EGFP B in frame with the C-terminal peptide to construct pMHES. 2.2.15 Cells were transfected with pcDNA6/myc-his-EGFP B and pMHES, and Balb/c mice were intravenously injected with pcDNA6/myc-his-EGFP B by tails, results revealed that both of the plasmids worked in 2.2.15 Cells and livers of Balb/c mice. Assuming gene of the IL-2 was inserted into pcDNA6/myc-his -EGFP B in frame with EGFP, myc and 6 × His, three-dimensional structure for this putative expression product was simulated using Modeller8V2, results revealed that IL-2, EGFP, myc and 6 × his did not interfere each other and octo- peptide linker owned certain flexibility. The results suggest that pcDNA6/myc-his-EGFP B may be useful as a genetic tool for mammalian cells and a vector for gene therapy.

ObjectiveTo investigate the role of δ-opioid receptor in remifentanil-induced N-methyl-D-aspartate (NMDA) receptor miniature excitatory postsynaptic currents (mEPSCs) in rat spinal dorsal horn neurons.MethodsMale 14-18 d old Wistar rats weighing 50-60 g were used in this study.The animals were anesthetized with intraperitoneal choral hydrate 400 mg/kg and sacrificed.Their lumbar segments of spinal cord (L1-S1 ) were immediately removed and sliced.Twenty-four slices were randomly divided into 4 groups ( n =6 each): control group (group C) ; glycine group (group G) ; remifentanil group (group R) and remifentanil + naltrindole(a δ receptor antagon) (group RN).Slices were cultured in artificial cerebrospinal fluid (ACSF) (group C) or incubated in ACSF containing glycine 0.24 μmol/L (group G) or remifentanil 4 nmol/L (group R) or remifentanil 4 nmol/L+ naltrindole 1 nmol/L (group RN) for 60 min.Whole cell patch clamp technique was used to measure NMDA receptor mEPSCs.ResultsThe amplitude and frequency of mEPSCs were significantly higher in group R than in groups C and RN ( P ＜ 0.01).There were no significant differences in amplitude and frequency of mEPSCs among gorups C,G and RN ( P ＞ 0.05).ConclusionActivation of δ-receptor can enhance NMDA receptor function in spinal dorsal horn neurons in rats which may be the mechanism of remifentanil-induced hyperalgesia.

Trimethoprim molecularly imprinted polymer (MIP) was prepared as the coating of stir bar sorptive extraction (SBSE) and applied to the trace analysis of trimethoprim and three sulfonamides in complex samples.The MIP-coating was about 21.5 μm thickness with the relative standard deviations (RSD) of 5.9％ (n=10).It was homogeneous and dense with good thermal and chemical stability.The extraction capability of the MIP-coating was 1.7 times over that of the non imprinted polymer (NIP) coating.The MIP coating exhibited selective adsorption ability to sulfonamides,triazines and methotrexate besides antibacterial synergists.The methods for the determination of trimethoprim and three sulfonamides by MIP-coated stir bar sorptive extraction coupled with HPLC were developed.It was successfully applied to the trace trimethoprim analysis in spiked urine and plasma samples.The linear range was 5 to 200 μg/L and the detection limit was 1.6 μg/L.The recoveries in urine and plasma samples were 84.5％ to 91.7％ with RSDs of 2.9％ -4.4％,71.9％ to 85.1％ with RSDs of 3.0％ -7.3％,respectively.The trimethoprim MIP-coated stir bar was also applied to the trace sulfonamides analysis in spiked milk sample.The linear range was 10-200 μg/L,the detection limit was within the range of 4.5-6.1 μg/L,and the recovery was 83.2％ - 110.2％ with RSDs of 4.1％ -8.0％.

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 observe the changes of immunological parameters and the effects of intravenous immunoglobulin (IVIG) in children with idiopathic thrombocytopenic purpura (ITP),schonlein-Henoch purpura(HSP) and mucocutaneoas lymph node syn-drome(kawasaki disease, KD).Methods T cell subsets and serum immunoglobulins and complements were measured with the flow cytometry and enhanced trubidimetric immunoassay. Routine therapy combined with IVIG. Results CD3+ CD4+ T cells were de-creased in children with ITP accompanied by increased serum IgG;Serum IgA and CD3+ T cells were increased in children with HSP; CD3+ CD8+ T cells were decreased in children with KD. Clinical features were markedly improved after treatment with IVIG. It was noted that the incidence of damage to coronary artery decreased after the use of IVIG. Conclusions T cell subsets, serum immunoglobulins should undergo the clinical routine examinations in ITP,HSP and KD. Early administration of IVIG might be important to improve disease prognosis and shorten its course of treatment in children with three kinds of immunology related diseases.