Objective To discuss whether ultrasonic attenuation dual calibration quantitative analysis may acquire highly repeatable checkout results. Methods With dual calibration phantom as references, under different total gain and depth compensation gain of B-mode ultrasonic scanner, test pending samples were tested. Different group images were collected and analyzed with self-developed ultrasonic attenuation quantitative analysis software. Results Test data had high repeatability under different total gain and depth compensation gain of B-mode ultrasonic scanner. Conclusion Ultrasonic attenuation dual calibration quantitative analysis can be used for quantitative analysis of B-mode ultrasonic images.

BACKGROUND:As a novel growth factor, human intestinal trefoil factor (TFF3) can promote cel growth and migration, and increase cel resistance to apoptosis, and it plays a great role in maintaining the mucosa integrity, mucosa protection and repairing the injured mucosa, also it has been closely related to the tumor growth and progression. With the function of mucosa repair, and as the tumor biomarker, TFF3 has a promising clinical application, but its definite interacting protein and molecular mechanism is stil unclear. OBJECTIVE:To construct and express the TFF3 recombinant protein with the tandem tag of StrepII-6×His in the target cel s for further purifying its interaction protein in the native condition based on the tandem affinity purification technique. METHODS:The DNA sequence for the tag (StrepII-TEV-6×His) and TFF3 as template was got by chemical synthesis and PCR amplification respectively. They were fused by the restriction enzyme XbaI site, and the tag sequence was located at the C terminus of TFF3 protein. TFF3-tag fusion gene was cloned into the pCDNA3.0 using EcoRI+HindIII, thus the TFF3-tag expressing vector pTFF3-C-StH was constructed and transfected transiently into gastric cel AGS by lipofectin. The recombinant TFF3-tag protein was expressed and detected by western blot assay. RESULTS AND CONCLUSION:The expressing vector pTFF3-C-StH for tandem affinity purification was constructed successful y, and was confirmed further by restriction enzyme analysis and sequenced. The recombinant TFF3-C-StH protein of TFF3-tag was expressed in the AGS cel , and showed specific antigenicity by western blot assay. Thus this work provides experimental base for further purification of the TFF3 interacting proteins.

Objective To investigate the role of platelet indices in the differentiation of thrombocytopenia caused by bone marrow diseases and thrombocytopenia caused by non-marrow diseases.Methods Mean platelet volume(MPV),platelet crit(PCT)and platelet size deviation width(PDW)of 76 othromboeytopenia(TP)patients,66 immune thrombocytopenia purpura(ITP)patients,8 secondary thrombocytocritpenia purpura(STP)patients,8 acquired pure megakaryocytic thrombocytopenia purpura(AATP)patients,140 malignant hematological diseases(MHD)(including acute leukemia,aplastic anemia and myelodysplastic syndromes)patients and 100 healthy people were respectively determined by blood cell analyzer Cell-Dyn 3700.Result Compared with control group,the MPV of the patients with MHD significantly decreased,and then the PDW of them increased.For patients with STP,the MVP and PDW markedly increased.The MVP of patients with AATP was normal,and PDW significantly increased.The MPV,PCT and PDW of ITP and TP showed marked difference(P＜0.01).In contrast with the control group,the MPV,PCT and the PDW of patients with hemopathy and patients with ITP were remarkably different(P＜0.01).Compared with the marrow TP and the non-marrow TP or control group,MPV and PDW showed significant difference(P＜0.01).During the effective treatment of patients with ITP,PLT increased and then MPV and PDW decreased.Conclusion The level changes of MPV and PDW,in particular MPV can primarily judge the cause of thrombocytopenia (MPV＜reference value as caused by bone marrow diseases but MPV＞reference value as caused by bone marrow diseases).In addition,during the effective treatment of patients with ITP,with changes of MPV and PDW present 5 days earlier than that of PLT,may used to observe the therapeutic effect.Myelodysplastic syndromedynamic observation of the platelet parameter helps for the differential diagnosis,judgment of the states of the diseases and observation of the effectiveness of therapy of ITP.

Objective To observe the clinical efficacy of acupuncture in treating subjective tinnitus.Method Seventy subjective tinnitus patients were randomized into a treatment group and a control group, 35 cases in each group. The control group was intervened by conventional medication, while the treatment group was by acupuncture in addition to the medication given to the control. The serum zinc level, copper level, and pure-tone hearing threshold were observed before and after intervention, and the clinical efficacies were compared.Result After intervention, the serum zinc and copper levels, and pure-tone hearing threshold were significantly changed in both groups (P<0.05). The serum zinc and copper levels, and pure-tone hearing threshold of the treatment group were significantly different from that of the control group after intervention (P<0.05). The total effective rate was 93.8% in the treatment group versus 87.9% in the control group, and there was a significant difference between the two groups (P<0.05).Conclusion Acupuncture plus medication is an effective way in treating subjective tinnitus.

OBJECTIVE:To observe clinical efficacy of extracorporeal shock wave lithotripsy combined with Paishi decoction in the treatment of kidney stone. METHODS:160 patients with kidney stone were selected and randomly divided into observation group and control group,with 80 cases in each group. Control group only received HB-ESWL-VG extracorporeal shock wave litho-tripsy;observation group additionally received Paishi decoction 300 ml,tid,for consecutive 1 week,on the basis of control group. Clinical efficacy was observed in 2 groups;the levels of serum inflammatory factor IL-2,IL-6,TNF-α,T lymphocyte subset CD3+,CD4+,CD8+ and CD4+/CD8+,erythrocyte immune function indicator C3b receptor rosette(C3bRR),immunity complex ro-sette(ICR)and tumor erythrocyte rosette(TER)level were detected in 2 groups before and after treatment. RESULTS:The effec-tive rate of observation group(98.75%)was significantly higher than that of control group(91.25%),with statistical significance (P<0.05). After treatment,the levels of IL-2,IL-6 and TNF-α in observation group was significantly lower than in control group, the levels of CD3+,CD4+,CD4+/CD8+,C3bRR,ICR and TER were significantly higher than control group,with statistical signifi-cance(P<0.05). Comparsion of CD8+between 2 groups was not statistically significant(P>0.05). CONCLUSIONS:The extracor-poreal shock wave lithotripsy combined with Paishi decoction can relieve inflammation reaction,enhance immune function and im-prove therapeutic efficacy.

Objective To establish a novel method to screen for anti-varicella-zoster virus (VZV) compounds with our previously generated reporter cell line for VZV, MV9G. MethodsMV9G cells were directly infected with cell-free virus of Oka vaccine strain (vOka) for 2 hours( CFV direct-infection) or cocultured with vOka-infected MeWo cells containing cell-associated virus for 48 hours (CAV co-culture) to promote expression of the reporter gene firefly luciferase. Antiviral compounds including heparin, mannose-6-phosphate( M-6-P), acyclovir( ACV ), resveratrol and roscovitine were added in the medium before or after the virus infection. Inhibitory effects( IC50 ) of the antiviral compounds were analyzed by comparing firefly luciferase activities of MV9G cells in the presence of antiviral compounds with those in the absence. Results Antiviral compounds inhibited luciferase activities of MV9G cells activated by CFV direct-infection and/or CAV co-culture in different levels. The reductions of luciferase activities statistically correlated with those of viral foci shown by immunostaining with a monoclonal antibody against VZV immediate early 62 antigens (IE62) in controls. Among these compounds, heparin, M-6-P, and 2.5 μmol/L of roscovitine inhibited CFV-activated more strongly than CAV-activated luciferase activities, whereas ACV and resveratrol inhibited CAV-activated more strongly than CFV-activated luciferase activities. Cell-associated ACV-resistant strains,Kanno and rOka YSR, activated luciferase activities of MV9G cells, too. However, the inhibitory concentrations (IC50) of ACV to the ACV-resistant strains were much higher than those to the ACV-sensitive strains,pOka and CaGu. ConclusionThe CFV direct-infection and CAV co-culture assays were useful to screen for antiviral compounds targeting the early and late phases of VZV infection, respectively. The VZV reporter cell-based assays may provide a simple, rapid, sensitive, and high-throughput method to screen for anti-VZV compounds.

To investigate the exon mutation of vitamin K-dependent gamma-glutamyl carboxylase (GGCX or VKDC) in patients with calcium oxalate urolithasis, renal cortex and peripheral blood samples were obtained from severe hydronephrosis patients (with or without calculi), and renal tumor patients undergoing nephrectomy. GGCX mutations in all 15 exons were examined in 44 patients with calcium oxalate urolithiasis (COU) by polymerase chain reaction (PCR) and denatured high pressure liquid chromatography (DHPLC), and confirmed by sequencing. Mutation was not found in all COU samples compared to the controls. These data demonstrated that functional GGCX mutations in all 15 exons do not occur in most COU patients. It was suggested that there may be no significant association between the low activity and mutation of GGCX in COU.

Objective: To explore the effect of umbilical cord blood stem cell transplantation on axon regeneration in spinal cord injury (SCl)in rats. Methods: The umbilical cord blood was collected and prepared into suitable concentration of CD34 positive stem cells. Thirty SD rats were divided into two groups randomly. One group served as control, another one was the treatment group. The models of spinal cord contusion injury were made by Allen's weight dropping method. One week later,the treatment group was transplanted with 10x105 umbilical cord blood stem cells with Hamilton micro-syringe at the sites of rostral and caudal to the lesioned zone respectively, while control group received just the same volume of PBS injection. Five rats in each group were sacrificed at 1 w, 2 w and 6 w after this operation. Histological and immunohistochemieal examinations including GAP-43 and NF200 were used to evaluate axon regeneration. Meanwhile, BBB motion scoring and inclined plane test were performed to assess the motion function changes of hindlimbs. Results: Compared to the control group, the area of cavity in the lesioned spinal cord region decreased significantly and the expressions of GAP-43 and NF200 increased markedly in cell transplantation group. Also the motion function had better restoration in the treatment group. Conclusion: Transplantation of umbilical cord blood stem cell may achieve both morphological and behavioral improvement for the injured spinal cord.

Objective: To explore and evaluate the biomechanic relationships between different depth of pedicle screw penetration with the sagittal plane reconstruction in thoracolumbar fracture. Methods:Six fresh cadaveric specimens of lumbar spine from L_1 to L_3 were used to make the model of thoracolumbar fracture. The system of universal spine system( USS )pedicle screw was adopted with the 6 mm diameter of screw. Each of two Schanz screws was implanted into the pedicles of L_1 and L_3 A canulated screw was fixed into the former of vertebral body in L_1 and L_3, and the distance of the two canulated screws was taken as the normal height. The axial loads were given while the pedicle screws were implanted at the depth of D1, D2 and D3, and the distance of the two canulated screw was measured as well as the distance was reduced to the normal height by axial load. The index measured included of the depth of pedicle screw penetration, the height of fractured vertebral body and afterload. Results: Along with the increasing of afterload, the height of injured vertebral body was increased accordingly, but the extent was different at three depth of pedicle screw penetration (D1, D2, and D3). While the injured vertebral body was reduced totally (reduced distance 0.00 mm), there was (2 630±13) g of afterload needed in Dl depth, and (2 339±61) g and(2 221± 164) g of afterload in D2 and D3 depth respectively. There was significant difference in distance between D1, D2 and D3 (P< 0.01), however, no significant difference between D2 and D3 (P> 0.05). Conclusion: There is a relationship in the depth of pedicle screw penetration, the capacity of reduction and sagittal plane reconstruction. The depth of pedicle screw had a significant effect on the capacity of reduction for the injured vertebral body, which would be the best option in biomechanics when the pedicle screw was implanted more than 1/2 pedicle or all of it.