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.

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 study the clinical efficacy of the injection Salvia (freeze-dried, abbr: Salvia powder needle), with fallopian tube recanalization on the treatment of oviduct obstructive infertility. Methods One hundred and ninety-five patients of oviduct obstructive infertility were enrolled in this study and divided into drug group and control group. Salpingography with modified catheter, recanalization and perfusion operation were performed in both two groups. Salvia powder, gentamicin, α-ACT and dexamethasone were injected into test group, while Gentamicin, α-ACT and dexamethasone were injected into control group. Recanalization rate of fallopian tube, rate of intrauterine gestation and rate of ectopic pregancancy were measured and compared between two groups. Results Twelve months after operation, recanalization of fallopian tube of test group and control group was 94.39% and 85.87%, respectively. Twenty-four months after operation, intrauterine gestation rate of test group and control group was 50.00% and 35.71%, respectively. Conclusion Salvia powder combined with modified recanalization operation is helpful to improve the rate of intrauterine gestation.

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.

Objective To explore the potential association of HLA-A alleles and genetic susceptibility with systemic lupus erythematosus (SLE). Methods Polymerase chain reaction-sequence specific primer (PCR-SSP) was used to analyze the distribution of HLA-A alleles among 106 patients with systemic lupus erythematosus and 122 healthy persons. Results Nineteen out of twenty-four kinds of HLA-A alleles were found from the specimens, including 18 kinds in SLE specimens, and 15 kinds in control specimens. Among them, HLA-A*11 allele was positively associated with SLE (RR = 2.4380, EF = 0.1502, ?2 = 12.2440, P = 0.0005, Pc = 0.0095). For A*01 and A*24, although the P values were less than 0.05, the Pc values were more than 0.05 (0.9462 or 0.2356, respectively). Conclusions The results indicate that HLA-A*11 may be the susceptible allele or may be closely linked with the susceptible genes in Chinese SLE patients.

Spider dragline silk is one of most perfect fibrous proteins in nature. As biomaterials, it has a wide application in tissue engineering due to its unique mechanical properties, good biocompatibility, slow degradation. In this paper, based on the highly repetitive sequence of spider dragline silk and with the introduced RGD peptide codons which involve cell adhesion, the DNA monomer sequence encoding RGD-spider dragline silk was synthesized, and then was used to construct the multimers by the strategy of "head to tail"; the multimers were ligated into prokaryotic expression vector pET-30a, and then the B121 (DE3) pLyS, were transformed the expression of recombinant protein was induced by the addition of IPTG. SDS-PAGE analysis shows that the molecular weight of products expressed here are 35KD and 60KD respectively in agreement with the desired. Western assay was used for determining the specification of products. Further, the purification process was groped for the producing of large quantity of synthetic proteins through high density fermentation.
Amino Acid Sequence ; Animals ; Base Sequence ; Biocompatible Materials ; Cloning, Molecular ; Fibroins ; biosynthesis ; genetics ; isolation & purification ; Molecular Sequence Data ; Oligopeptides ; biosynthesis ; isolation & purification ; Polymers ; isolation & purification ; Protein Engineering ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Spiders

Objective: To compare the effect of scalp acupuncture and scalp acupuncture plus acupuncture exercise therapy (AET) on walking ability in children with spastic cerebral palsy (CP). Methods: A total of 60 spastic CP children with gross motor function classification system (GMFCS) grades Ⅰ-Ⅲ were divided into a control group and an observation group by the random number table method, with 30 cases in each group. Both groups were treated with the same conventional rehabilitation and scalp acupuncture therapy for CP. The control group received conventional rehabilitation first and then scalp acupuncture. The observation group received AET, which was to receive the conventional rehabilitation and scalp acupuncture simultaneously. Before and after treatment, the clinical efficacy was evaluated by the modified Ashworth scale (MAS) score, scores of dimensions D and E of the gross motor function measure (GMFM) scale, walking speed, and walking distance. Results: During treatment, there were 2 dropouts in the observation group. After 3 courses of treatment, the MAS scores in both the control group and observation group decreased compared with the same group before treatment (P<0.05), and the scores of dimensions D and E of the GMFM, walking speed, and walking distance were increased (P<0.05); the between-group comparison showed that the MAS score in the observation group was lower than that in the control group (P<0.05), and the scores of dimensions D and E of the GMFM, walking speed, and walking distance in the observation group were higher or longer than those in the control group (P<0.05). Conclusion: W ith the same treatments, scalp acupuncture combined with AET is superior to the conventional scalp acupuncture method in reducing lower-limb muscle tone, improving standing balance ability, and walking stability in children with spastic CP.

Objective To investigate the expression of angiopoietin 1 and 2(Ang1,Ang2)in diffuse large B-cell lymphoma(DLBCL)and its survival prognosis analysis.Methods Peripheral blood was collected from 32 patients with initially diagnosed DLBCL and 30 healthy volunteers(the health control group),and the serum Ang1 and Ang2 levels were detected by ELISA.The biopsy tissues from DLBCL patients and the resec-ted normal lymph node biopsy tissues in neck surgery of 25 patients with non-tumor lesions(the control group)were taken as the specimens.The immunohistochemistry was used to detect the expression of CD34 in lymphoid tissues and the microvessel density was calculated;the statistical analyses were carried out by combi-ning with the clinical characteristics of the DLBCL patients and the follow-up data.Results The serum Ang1 level in initially diagnosed DLBCL was higher than that in the healthy control group[(36.22±9.12)ng/mL vs.(30.92±13.37)ng/mL],and the serum Ang2 level in initially diagnosed DLBCL was higher than that in the healthy control group[(28.42±10.78)ng/mL vs.(23.81±3.68))ng/mL],and the differences were sta-tistically significant(P＜0.05).CD34 was highly expressed in the lymphoma tissues of the patients with DL-BCL,whereas CD34 in normal lymph node tissues was weakly expressed;the microvessel density count in the DLBCL group was increased compared with the normal lymph node group(25.5±4.4 vs.13.2±3.0,P＜0.05).The Ang1 expression level had no significant correlation with the gender,age,IPI score,clinical stage and COO subtype.The Ang2 expression was correlated with the clinical stage,Ang2 was lowly expressed in the patients with stage Ⅰ-Ⅱ DLBCL,while which was highly expressed in the patients with stage Ⅲ-Ⅳ(P＜0.05);Ang2 had no significant correlation with the gender,age,IPI score and COO subtype.The OS time had no significant difference between the patients with Ang1 low expression and the patients with Ang1 high expression(25.86 months vs.23.11 months,P=0.722);the OS time in the patients with Ang2 low ex-pression was significantly higher than in the patients with high Ang2 expression(32.24 months vs.17.66 months,P=0.002).In the univariate analysis,Ann Arbor stage,IPI score and Ang2 all were the prognostic factors affecting the patients'OS,while gender,age and Ang1 had no significant correlation with the patients'OS.In Cox multifactorial analysis,the Ann Arbor stage,IPI score and Ang2 all were the independent prog-nostic factors affecting the patients'OS.prognostic factors.Conclusion Ang1 and Ang2 are highly expressed in DLBCL patients and promote lymphoma angiogenesis;Ang2 affects the survival prognosis of DLBCL patients.

Objective:To predict the mechanism of Danggui Buxue Decoction for anti-myocardial ischemia-reperfusion injury and "treating different diseases with the same method" in ischemic stroke based on network pharmacology and molecular docking.Methods:The active components and targets of Danggui Buxue Decoction were screened by retrieving the database of TCMSP and literature; the corresponding targets of myocardial ischemia-reperfusion injury and ischemic stroke were found by OMIM and GeneCards database; the intersection targets of Danggui Buxue Decoction and disease were obtained by using Venny diagram, and the common target network and protein-protein interaction network were constructed by Cytoscape 3.7.1 software and STRING database. The GO and KEGG pathways were enriched by David Database, and the Bio GPS database was used to obtain the tissue distribution information of the key targets. The molecular docking technology was used to verify the results.Results:There were 21 active components in Danggui Buxue Decoction, 181 effective targets and 93 cross targets with diseases. The key components were quercetin, Kaempferol, β-sitosterol, formononetin and isorhamnetin. The key targets were AKT1, TNF, IL6, IL-1β and VEGFA. The enrichment results showed that the main action pathways were fluid shear force and arteriosclerosis, lipid and arteriosclerosis, AGE-RAGE signal pathway in diabetic complications, and the core targets were mainly located in the medullary cells, dendritic cell, smooth muscle, prostate, thyroid and other tissues. The results of molecular docking showed that quercetin had the best binding effect to IL-1β, while isorhamnetin had the best binding effect to IL-1β.Conclusion:Danggui Buxue Decoction is against myocardial ischemia-reperfusion injury and ischemic stroke through hemodynamics, lipid metabolism, inflammatory reaction, oxidative stress, immune reaction and cell apoptosis, plays the role of "treating different diseases with the same method".

Humans ; Clavicle ; Fractures, Bone/surgery*