The working principle and application characteristics of ECG machine were introduced in detail from such aspects as the development history,structure,classification,new usage and procurement principle.The objective is to direct new equipment procurement and facilitate clinical application.

ObjectiveExpression and purification of human tumor necrosis factorα(hTNFα)fusion protein with a stretch of six consecutive histidine residues(6×His) in E. coli.MethodshTNFα fusion proteins with 6×His at N and C terminus were expressed by using E. coli expression vectors pET- 28a(+) and pET-22b(+). The His6 tag allows the expression fusion protein purified in one step by immobilized metal Ni2+ chelation affinity chromatcgraphy in native state. Results The two construct expression vectors were expressed in E. coli respectively, the former with high level as insoluble protein, account for 45% of the total bacteria proteins and not purified; the later 8% as soluble protein, and characterized by SDS- PAGE, Westren-blot. By using affinity chromatography through Ni2+ - IDA Sepharose 6B, 100ml induction culture had 0.4mg hTNFα-6×His fusion proteins. Its purifity reached 90 %. ConclusionThe purification expression product can possess TNF activity and reach 5.42×104U/mg.

Objective To produce bioactive human bone morphogenetic protein-4 by Escherichia coli genetic engineering and investigate the effect of the product, recombinant BMP-4, on the bone marrow stem cells. Methods cDNA of human morphogenetic protein-4 mature peptide was obtained by RT PCR from tissue of human placenta. The gene was constructed in the pET-22b(+) expression vector and expressed in Escherichia coli BL-21 after transformation and induction by IPTG. The harvested protein was proved to be bioactive by inducing ectopic bone information in mouse thigh. The protein was applied to induce the cultured bone marrow stem cell. Shape change of the cell, ability of ALP (alkaline phosphatase) and concentration of OC (osteocalcin) were investigated. Results SDS-PAGE revealed a new protein band that located in position of 14?103 after 4 hours induction, the new protein made 15% of total bacteria protein, the rhBMP-4 could induce the cultured bone marrow stem cell of New Zealand rabbit to differentiate into osteoblasts and form calcified node. The ability of ALP and concentration of OC of tested group increased significantly than that of the control group. Conclusion The bioactive rhBMP-4 can be produced by Escherichia coli genic engineering, this protein can induce bone marrow stem cells to differentiate into osteoblast cells.

BACKGROUND:Because of their immunological properties, bone marrow mesenchymal stem cells transfusion is developed as a new treatment for refractory graft-versus-host disease. OBJECTIVE:To analyze the safety and curative effect of bone marrow mesenchymal stem cells transfusion on treating different organ damages in graft-versus-host disease after al ogeneic hematopoietic stem celltransplantation. METHODS:Eight patients with malignant hematologic disease were included in this study. The patients developed severe steroid-resistant graft-versus-host disease after al ogeneic hematopoietic stem celltransplantation and received transfusion of mesenchymal stme cell(1×106 of immunosuppressive agent. RESULTS AND CONCLUSION:For the total y eight patients, six got response (two cases of complete remission, and four cases of partial remission) and two showed no remission. Four of five cutaneous damages were ameliorated and one showed no effect. For three cases of oral graft-versus-host disease, two acquired complete remission and one showed partial remission. Two cases of liver graft-versus-host disease and two cases of astro-intestinal graft-versus-host disease obtained complete remission. No response was displayed to three cases of ocular graft-versus-host disease, one case of bronchiolitis obliterans, and one case of urinary graft-versus-host disease. In the median fol ow-up of 28 months (7-62 months), three patients developed posttransplant lymphoproliferative disorders within 3 months after mesenchymal stem cells transfusion. Administration of mesenchymal stem cells is safe for treatment of severe graft-versus-host disease after al ogeneic hematopoietic stem celltransplantation. Mesenchymal stem cells transfusion may be a promising/kg) together with the primary therapy therapy for refractory cutaneous , astro-intestinal, liver and oral graft-versus-host disease but not for pulmonary, ocular and urinary graft-versus-host disease. Whether mesenchymal stem cells transfusion is associated with posttransplant lymphoproliferative disorders needs more case data.

Objective To explore regional anatomical features of fascia and spaces related to complete mesocolic excision (CME)dur-ing laparoscopic right hemicolectomy.Methods Observe and describe the regional anatomical features of related mesenterium,fascia and spaces through autopsy and somatoscopy.Results Superior mesenteric vein is the anatomic landmark in CME with medial access for laparo-scopic right hemicolectomy.Right mesocolon and ileal mesentery are the main mesenterium,and the fascia contains the prerenal fascia and the pancreatic fascia.The right retrocolic space and the colon transversum space are two important anatomical spaces,and their fusion fascia space served as a natural surgical plane.Conclusion There is a natural surgical plane which made of mesenterium,fascia and spaces be-tween mesocolon and prerenal fascia in CME during laparoscopic right hemicolectomy,and the surgery is feasible.

Objective To study the causes, emergency managements and preventative measures of intraoperative rupture during embolization of intracranial aneurysms. Methods One hundred and thirty-two patients with intracranial aneurysms and having performed embolization were enrolled. Intraoperative rupture occurred in 6 patients during embolization of intracranial aneurysms. The causes were analyzed and emergency managements were reviewed. Results The causes were as follows: intracranial aneurysms were punctured by microtubular in 2 patients, by spring coil in 1 patient and by guidewire in 1 patient, and intracranial aneurysms were attributed to stent break in 1 patient and blood pressure fluctuation in 1 patient. The rapid embolization were performed, 1 patient underwent craniotomy evacuation of hematoma, 1 patient underwent lateral ventricular drainage, 2 patients underwent lumbar cistern drainage, and 2 patients did not receive any processing. At the discharge from hospital, 2 patients recovered well, 1 patient showed mild paralysis, 1 patient had severe disability, and 2 patients died. Conclusions Intraoperative rupture during embolization of intracranial aneurysms is the life-threatening complication and mainly relates to operation, structure of aneurysm, blood pressure fluctuation during operation. The prognosis depends on the extent of bleeding and appropriate treatment without delay.

Objective To observe the clinical efficacy ofErxian Decoction combined with elcatonin for the treatment of postmenopausal osteoporosis.Methods Eighty cases of female patients with postmenopausal osteoporosis were randomly divided into treatment group and control group, 40 cases in each group. The control group received intramuscular injections elcatonin treatment, and the treatment group received oralErxian Decoction on the basis of the treatment of the control group, for 12 weeks. The main clinical symptom score, VAS score and bone mineral density (BMD) of L2-L4 and in the tibia of the two groups were observed before and after treatment to assess the therapeutic effect.Results After treatment, VAS scores of the two groups were significantly lower (P<0.05). The total effective rate of the treatment group was 92.5% (37/40); the control group was 75.0 (30/40), with statistical significance (P<0.05). The treatment group was significantly better than the control group in main symptom scores, especially in improving body symptoms (P<0.05). BMD of the treatment group was significantly higher than the control group.ConclusionErxian Decoction combined with elcatonin therapy for postmenopausal women with osteoporosis can significantly improve clinical symptoms and increase BMD.

Objective To explore the anti-tumor effects of Egr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy in mice bearing H22 hepatocarcinoma and its mechanism. Methods The recombinant plasmid pcDNAEgr-IFNγ mixed with liposome was injected into tumor. 48 h later, 370 kBq 125I-UdR was injected into tumor. The tumor growth rates at different times were observed. After 3 d gene-radionuclide therapy, the concentration of IFNγ in cytoplasm of H22 cells and cytotoxic activities of splenic CTL of the mice in different groups were examined. Results The tumor growth rates of pcDNAEgr-IFNγ +125 I-UdR group were obviously lower than those of control group, 125I-UdR group and pcDNAEgr-1 +125I-UdR group 6-15 d after gene-radionuclide therapy. IFNγ protein was found in cytoplasm of H22 cells in PcDNAEgr-1FNγ+125I-UdR group after 3 d gene-radionuclide therapy. Cytotoxic activity of splenic CTL in pcDNAEgr-IFN7 + 125I-UdR group was significantly higher than that in the other groups (P<0.01). Conclusions The anti-tumor effects in vivo of pcDNAEgr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy are better than those of 125I-UdR therapy.

AIM:To investigate the expression of aplasia rashomolog member I ( ARHI) gene in acute myeloid leukemia cells (AML) and to study the effects of ARHI on the growth of AML cell line U937.METHODS:The mRNA ex-pression of ARHI in AML cells, 293FT cells, AML primary cells and healthy volunteer blood cells were detected by RT-PCR.After transfection with the MSCV-IRES-GFP-ARHI plasmid to the U937 cells, the growth curve was analyzed by MTT assay.U937 cells were re-suspended by fresh medium and cultured for 24 h, then the cell cycle distribution and ap-optotic rate were determined.RESULTS:The mRNA of ARHI was positively detectable in 293FT cells and healthy volun-teer blood cells instead of AML cell line and AML primary cells.The growth curve showed that cell viability in U937 cells with high expression of ARHI (U937-ARHI) was lower than that in the control cells (U937-GFP) on 6th~8th day.The ratio of G2/M phase and apoptotic rate in the U937-ARHI cells were increased compare with control group ( P<0.05 ) . CONCLUSION:The mRNA level of ARHI is low in AML cells.High expression of ARHI gene in U937 cells inhibits cell growth, arrests the cells at G2/M phase and induces apoptosis.

Objective To compare and analyze the effect of minimally invasive percutaneous nephrolithotomy compare with traditional open surgical in treatment of kidney stones outcome. Methods Selected 120 patients with kidney stones from January 2009 to January 2014 in our hospital, all patients had complete clinical data, included 60 cases of renal sinus traditional lines pelvis lithotomy, established as the control group, and the remaining 60 routine minimally inva-sive percutaneous nephrolithotomy surgery, set up minimally invasive group, the operative time, blood loss, postoperative fistula time and hospital stay and complications were compared between two groups. Results 60 patients with invasive group homogeneity of puncture success and smooth gravel, stone successful in 58 cases, the success rate was 96.7%(58/60). Intraoperative complications pneumothorax, colon injury does not appear, one-time stone clearance rate was 80.0%(48/60). Stable vital signs after surgery, no nephrectomy, severe complications wound fistula, pneumothorax oc-curred. 4 cases of postoperative urinary tract infection sustained high fever, anti-infection cure. 1 case duodenal injury control group intraoperative peritoneal injury in 2 cases, 2 cases of postoperative fever, intestinal fistula, urinary tract infection in 8 cases. Operative time, blood loss, postoperative fistula compare the time and duration of hospitalization, of observation group improved significant than the control group, the difference was statistically significant (P<0.05 or P<0.01). Conclusion Minimally invasive percutaneous nephrolithotomy in the treatment of kidney stones with less trau-ma, less bleeding, fewer complications, shorter hospital stays and can be worthy of promotion and application.