BACKGROUND: It is a new tendency to treat central nervous system injury or tumor therapy using the combination of seed cells and gene therapy.OBJECTIVE: To observe the dose-relationship between transfection and expression of rabbit bone marrow mesenchymal stem cell (BMSCs) with adenovirus vector carrying green fluorescent protein (Ad-GFP), and to study its effects on cell biological properties, in addition, to explore the feasibility of using Ad-GFP vector to construct gene modified BMSCs.DESIGN, TIME AND SETTING: A randomized grouping, contrast observation. The experiment was performed at the Southern Medical University between August 2008 and March 2009.MATERIALS: New Zealand white rabbits, irrespective of genders, weighing 2.0-3.0 kg, were selected.METHODS: BMSCs were separated and cultured in vitro, and then the cell immunophenotypes were detected by flow cytometry.The adenovirus was obtained by packaging 293 cells and was used to transfect BMSCs with various liters (1 ×-10~3-1×10~(10) PFU/mL).Cytometry was used to analyze the transfection efficiency.MAIN OUTCOME MEASURES: Cell morphological changes were detected under an invert microscope. The cell proliferation was detected by CCK8 kits. BMSCs transfected with Ad-GFP were induced differentiating into neuron-like cells by adding of β-mercaptoethanol.RESULTS: The surface markers of 3-6-generation BMSCs were negative to CD34 and CD45, but positive for CD29 and CD44.When the virus titers were 1 ×10~7 PFU/mL, the transfection rate was 55%, which were 85% when the virus titers were 1 ×10~9 and1×10~(10) PFU/mL. However, cell pathology phenomenon occurred when the virus titer was 1 ×10~(10) PFU/mL. The fluorescence was strongest expressed at day 7, and it still can be seen at day 28. The BMSCs trasfected with Ad-GFP could differentiate into neuron-like cells under induction of p-mercaptoethanol, with positive neuron-specific enolase.CONCLUSION: Ad-GFP with suitable titers can infect BMSCs effectively with little influence on the biology property or differentiation function. BMSCs can serve as seeds cell in gene therapy field when utilizing ad-GFP vector system.

Objective To investigate the curative effect of using new irrigation pump in acute hematogenous osteomyelitis.Methods Forty patients with acute hematogenous osteomyelitis were divided into the control group and observation group,20 cases in each group.The control group adopted the conventional lavage for drainage,while the observation group used self-made irrigation pump.Then the clinical curative effect and cure time were compared between the two groups.Results The total effective rate in the observation group was 100.00 ％,which was significantly higher than 80.00 ％ in the control group(P＜0.05),and the difference was statistically significant(P＜0.05).The cure time in the observation group was(10.13 ± 8.03)d,which was significantly shorter than (18.14-±5.43)in the control group,and the difference was statistically significant(P＜0.05).Conclusion The curative effect of new type irrigation pump in acute hematogenous osteomyelitis is more acurate than that of conventional lavage for vacuum darianage.

BACKGROUND:Many scholars have developed a variety of dynamic elastic spine fixator. After biomechanical research, animal experiments and clinical application found that no one elastic spine fixator was general y recognized clinical y. OBJECTIVE:To compare the stress difference between“X”-shaped spine dynamic fixation and traditional pedicle screw fixation. METHODS:Three-dimensional finite element models of“X”-shaped spine dynamic fixation and traditional pedicle screw fixation were established according to adult spine imaging data. Mechanical differences in vertical compression, flexion, extension, lateral bending and rotation were compared between the two groups. RESULTS AND CONCLUSION:The stress at vertical compression was lower than that at flexion, extension, lateral bending and rotation in both groups. The stress at“X”-shaped spine dynamic fixation mainly focused on“X”-shaped connecting rod, but the stress of traditional pedicle screw fixation mainly focused on conjunction of screw-rod. Moreover, the stress of the screw of“X”-shaped spine dynamic fixation was significantly less than that of traditional pedicle screw fixation (P<0.001). These results suggest that“X”-shaped spine dynamic fixation system can share stress of screws and reduce the postoperative stress concentration compared with traditional pedicle screw fixation.

Abstract?AlM:To analyze of the clinical features of acute retinal pigment epitheltis ( ARPE) .?METHODS: The clinical data of 36 ARPE patients ( 40 eyes) attending this center from January 2008 to January 2014 were reviewed retrospectively. Of them, 21 patients (58.3%) were male (male :female=1:0. 71). The mean age was 40. 92±7. 13 years old (range:17~60y). The mean best-corrected visual acuity (BCVA) was 0. 50±0. 26 with a range of 0. 3 ~ 1. 0. Thirty-two patients were unilateral cases. All the patients were examined for BCVA, funds photography, fluorescein fundus angiography ( FFA ) , optical coherence tomography ( OCT) . FFA was shown as three types: type ▏ to multiple black light or grape variety fluorescent spot; Type II for l lesions visible fluorescence leakage; Type Ⅲ lesions with choroid neovascularization ( CNV ) . OCT was the following three forms: multiple RPE lesions layer reflection intermittent, proliferation ( type ▏); pigment epithelial detachment with limitations neural epithelium ( typeII);types l and ll with CNV ( type Ⅲ) .?RESULTS: Ocular fundus showed that the lesions were multiple dark-gray spots with a dark circumscribed area at the macular or nearby in all 40 eyes. FFA showed:21 eyes were type ▏, 17 eyes were type II and 2 eyes were typeⅢ, BCVA between type ▏ and type II was statistically significant (P<0. 05), the same was between type 芋. BCVA between different cases in the same type and between type II, Ⅲ, was no statistical difference ( P>0. 05). OCT showed 21 eyes wwere type ▏, 17 eyes were type II and type Ⅲ 2 eyes. BCVA average between type▏ andIIwas statistically significant (P<0. 05). The mean BCVA was no statistically significant difference between type II and Ⅲ patients (P>0. 05).?CONCLUSlON:ARPE fundus demonstrated the multiple dark gray discrete lesions, the degree of visual impairment related with the presence of pigment epithelial barrier and lesion location. OCT and FFA characterized three types. FFA is shown asblack light orgrape variety fluorescent spot, and is the basis of diagnosis. OCT can display the lesions organization form of each layer clearly. lt plays a more and more important role in the diagnosis and differential diagnosis of ARPE.

OBJECTIVETo investigate the dynamic changes of the epididymal size 1 year after vasectomy.

METHODSFifty male volunteers received vasoligation. Before and at 1, 2, 3, 6, and 12 months after operation, we measured the size and detected the internal echoes of the epididymis using color Doppler ultrasonography.

RESULTSThe bilateral epididymides were both thickened post-operatively in all the 50 cases, with statistically significant differences between the baseline and the 1st month, the 1st and the 2nd month, the 2nd and the 3rd month, or the 3rd and the 6th month after surgery (all P < 0.01), but not between the 6th and the 12th month (P > 0.05).

CONCLUSIONWithin 6 months after vasectomy, the bilateral epididymides manifested a progressive thickening, but basically restored their balance of secretion-absorption after 6 months.

Epididymis ; diagnostic imaging ; pathology ; physiology ; Humans ; Male ; Organ Size ; Postoperative Period ; Time Factors ; Ultrasonography, Doppler, Color ; Vasectomy

Objective To detect the levels of five trace elements in whole blood of patients with Keshan disease(KSD) and dilated cardiomyopathy(DCM) and explore their role in the pathogenesis of KSD.Methods One hundred and four patients with chronic KSD were selected from Keshan diseased areas in Shandong,Sichuan and Inner Mongolia.Thirty patients with DCM were selected from Qilu Hospital of Shandong University,Jinan Central Hospital,The First People's Hospital.Ninety-one healthy people from KSD endemic areas and 39 healthy people from Jinan were selected as endemic healthy controls and non-endemic healthy controls,respectively.Blood samples were collected to determinate the level of selenium (Se),copper (Cu),zinc (Zn),chromium (Cr) and manganese (Mn) with fluorescence method and atomic absorption spectrometry,according to the principle of informed consent.Results The level of Se,Zn and Cr of KSD group[(36.0 + 4.9)μg/L,(22.73 + 4.62)mg/L,(0.56 + 0.17)mg/L] was significantly lower than that of non-endemic healthy controls [(56.4 ± 6.8)lμg/L,(25.35 ± 4.44)mg/L,(0.71 ± 0.17)mg/L,all P ＜ 0.05],but the level of Cu of KSD group[(0.95 ± 0.24)mg/L] was significantly higher than that of non-endemic healthy controls[(0.73 ± 0.13) mg/L,all P ＜ 0.05].The level of Se and Cr of KSD was significantly lower than that of endemic healthy controls[(54.5 ± 5.4)μg/L,(0.87 ± 0.02)mg/L,P ＜ 0.05],and Cu was significantly higher than that of endemic healthy controls[(0.66 ± 0.02)mg/L,P ＜ 0.05].The level of Cu and Zn of KSD was significantly lower than that of DCM [(1.21 ± 0.23)mg/L,(27.09 ± 7.10)mg/L,all P ＜ 0.01].The level of Se and Cr of DCM group[(39.6 ± 3.5)μg/L,(0.58 ± 0.14)mg/L] was significantly lower than that of non-endemic healthy controls(all P ＜ 0.01),but Cu[(1.21 + 0.23)mg/L] was significantly increased (P ＜ 0.01).Compared with non-endemic healthy controls,the level of Se of endemic healthy control group was significantly decreased (P ＜ 0.01),while Cu was significantly increased (P ＜ 0.01).Se,Zn and Cr level of KSD decreased gradually following elevated heart function level,but the level of Cu gradually increased.Conclusions The metabolism of Se,Cr,Cu and Zn is unbalanced in KSD patients,whose Se level is still lower than that of people in non-endemic areas.The change of Se,Cr,Cu and Mn level between KSD and DCM is consistent.

OBJECTIVETo investigate the clinical characteristics of children with Castleman's disease and to improve doctors' awareness of this disease.

METHODSClinical characteristics of 5 cases with Castleman's disease were observed and analyzed and relevant reports in literature were reviewed.

RESULTS(1) All the five patients' histories were long, and the first symptoms of them were painless lymphnode enlargement, and all of them were at school age; 3 patients' abdominal lymphnodes were enlarged, mediastinum lymphnodes enlarged in 3 cases, cervical lymphnodes were involved in 3 cases; (2) The clinical subtypes: the disease in 3 cases was localized Castleman's disease (LCD), all of their pathological subtype was hyaline vascular variant (HV). The rest of them were multicentric Castleman's disease (MCD), whose pathology was plasma cell variant (PC), and both of them had a febrile symptoms; (3) The white blood cells, C-reactive protein and ferritin levels were all elevated to different extents. Four of them had viral infections, and their cellular immune function was abnormal; (4) The LCD patients' prognosis was good after the complete resection. There is no standard therapy for MCD, the available therapies include antiviral, immune modulatory regimens, CD20 B cell monoclonal antibody and chemotherapy, but the prognosis was worse than that of LCD.

CONCLUSIONSCastleman's disease is rare in children, which can be misdiagnosed because it has no specific manifestations. The prognosis depends on the subtype.

Castleman Disease ; diagnosis ; Child ; Female ; Humans ; Male ; Prognosis ; Retrospective Studies

Apx IV, a forth RTX toxin indentified in Actionbacillus pleuropneumoniae recently, is expressed by all A. pleuropneumoniae regardless the serotypes and inducible only in vivo toxin, so it is the optimal to develop species-specific and differentiated diagnostic assay. Here the 2445bp DNA fragment of apxIVA gene of A. pleuroneumoniae was amplified and fused in-frame to the downstream of the T7 promoter and 6 His Tag of the prokaryotic expression vector pET-28b. The construct was transformed into E. coli BL21(DE3). After induction by 1.0 mol/L IPTG, a recombinant protein about 90 kD in size, designed as ApxIVAN, was detected, which was present as inclusion bodies and reacted specifically with swine antisera to the APP-serotype-1 by dot-blot. An indirect ELISA (ApxIVA-ELISA) was developed using purified recombinant ApxIVAN from the inclusion bodies as described previously, which had excellent specificity to A. pleuroneunoniae. Using the ApxIVA-ELISA, the ApxIV antibodies were not detected in the inactivated APP bacterins vaccinated pigs, but were detected in A. pleuropneumoniae serotype 1, 2 and 7 infected pigs and mice. These results suggested that ApxIVA-ELISA can be used not only to detect all serotypes of APP, but also to differentiate the naturally infected and inactivated vaccine immunized pigs.
Actinobacillus Infections ; diagnosis ; microbiology ; veterinary ; Actinobacillus pleuropneumoniae ; genetics ; immunology ; metabolism ; Bacterial Proteins ; genetics ; immunology ; metabolism ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; veterinary ; Gene Expression ; Genes, Bacterial ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

The HLA system was discovered by virtue of the fact that it was polymorphic. The impetus for its discovery was the search for polymorphic antigens to match for transplantation, by analogy with the human red cell blood groups. The most usually DNA method of HLA typing is sequence specific oligonucleotides (SSO) and PCR sequence specific primers (SSP). SSO technique is perfectly suited for analyzing large number of samples, it is not suitable for individual or small numbers. The SSP method is ideal for typing individual samples, but it is costly and requires high capacity thermal cycles for larger numbers of samples. To set up a simple, quick, cheap and high resolution DNA method, were collected sixty-three cord blood samples from Guangzhou Cord Blood Bank, got DNA from blood by the traditional guanidine hydrochloride distillation method. Each sample was simultaneously typed by SSOP, PCR-SSP and reverse dot-blot hybridization (RDB) methods. All of typed is success. The results of three DNA methods are consistent each other. 60 HLA-DRB1 alleles could be accurately distinguished with the RDB method. Our results show that RDB method is a simple, quick, cheap and high resolution method for HLA-DRB types. It can be used in any HLA typing.
Fetal Blood ; immunology ; metabolism ; Genotype ; HLA-DR Antigens ; genetics ; Histocompatibility Testing ; methods ; Humans ; Nucleic Acid Hybridization ; methods

Objective To investigate the phenotypes and genotypes of a hereditary protein C(PC) deficiency pedigree.Methods Imrnunoassay(ELISA)was used for PC antigen and PS antigen; Immunoturbidimetry assay was used for measuring AT antigen;Chromogenic substrate assay was used for measuring the activity of PC,PS and AT in Sysmex 1500 automatic Blood Coagulation Analyzer.Polymerase chain reaction(PCR)for amplification of the fragment of each exon and side sequences of PC gene in 10 members of the 3 generations;Direct DNA sequencing was used to examine the mutation site.Results Among 10 members of the 3 generation pedigree,8 of them had a PC:Ag level of 1.06-1.92 mg/L(normal references 3.00-6.00 rag/L),the activity of PC was between 41% and 67%(normal references 70%- 140%),which was significantly lower than the normal references while the levels of PS:Ag,PS:A,AT:Ag and AT:A were all within normal range.DNA sequencing analysis showed that there was a G to T mutation in exon IX of the PC gene at 12 918 position in 8 members.This mutation resulted in the substitution of terminator TGA for TGG which encoding tryptophan at 372 amino acid.There was a polymorphism in 2 405C/ T,2 418A/G,2 583A/T in the promotor area.Conclusions This pedigree is a type I hereditary protein C deficiency.There is a G12 918T mutation in exon IX of PC gene.This mutation is reported for the first time and there is a polymorphism in 2 405C/T,2 418A/G,2 583A/T in the promotor area.