Objective To analyze the characteristics of proximal deep vein thrombosis (DVT) after total joint arthroplasty.Methods 40 cases of proximal DVT were collected from May 2007 to October 2016 which were all diagnosed by lower limb venography or sonography.There were 9 males and 31 females aged from 38 to 86 years,average 67 years,and body mass index was from 16.94 to 31.25kg/m2,with an average of 24.3 kg/m2.29 cases of THA patients and 11 cases of TKA patients.The anatomical distribution,size,radiological performance,local physical findings were recorded and multivariable analysis was performed to evaluate the relationship between distribution of proximal DVT and related parameters including age,gender (male/female),side of leg(left/right),surgery type(knee and hip),preoperative diagnosis(femoral neck fracture/other disease),time of diagnosis with DVT (during hospitalization/after discharge).Results Among 40 proximal DVT cases,31 cases diagnosed by phlebography and 9 cases were diagnosed by ultrasound.2 cases were isolated proximal DVT while 38 cases connected with distal DVTs.9 cases of blood clots in the proximal and distal thrombosis connected discontinuously.There was no isolated proximal DVT in TKA group and 10 cases (90.9％) of blood clots in the proximal and distal thrombosis connected continuously while there were 2 cases (6.9％) isolated proximal DVTs in THA group and 21 (77.8％) of blood clots in the proximal and distal thrombosis connected discontinuously,and there was no statistic significant difference between the two groups.The average length of proximal was 8.85±9.3 1cm (range from 2-35cm) with 5.0±2.05 cm in TKA group and 10.31 ± 10.55 cmin THA group.A significant difference was found between the 2 groups (P=0.014).13 cases located in femoral veins and upper venous region which were all from THA group.There was a significant difference in the distribution between surgery type and preoperative diagnosis.However,there was no significant difference between other potential factors and distribution of proximal DVT.25 cases accepted the evaluation of symptom of DVT before scanning.However,no significant difference were found in edema,VAS score,Homans sign,Neuof sign between 9 TKA and 16 THA cases.Conclusion Proximal DVTs after knee and hip arthroplasty are more preferred to connect with distal DVT continuously.Surgery type and preoperative diagnosis have a significant effect on the distribution of proximal DVT.Proximal DVTs in THA and patients with preoperative femoral neck fracture could be more easily involving femoral veins and upper parts.

OBJECTIVETo investigate the relationship between retroviruses and autoimmune diseases, to clone the novel retroviral NP9 gene from human endogenous retrovirus (HERV), and to construct its expression vector.

METHODSThe viral NP9 gene was amplified and cloned by RT-PCR and T-A clone techniques, and its sequence was determined with Perkin-Elmer 377 DNA Sequencer. The amplified viral NP9 gene was subcloned into the prokaryotic express vector pQE30. The recombinant plasmids were identified by restriction endonuclease digestion and sequencing. The recombinant pQE30-NP9 protein was expressed in M15 host cells under the IPTG induction and showed with SDS-PAGE,and the corresponding NP9 viral protein was identified with Western blot analysis.

RESULTA specific band of 250 bp was amplified using RT-PCR from total RNA of peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) and confirmed as the NP9 gene via T-A clone and DNA sequencing analyses. SDS-PAGE profile showed a clear protein band with a relative molecular weight 9 kD in the IPTG-induced samples, which was confirmed as viral NP9 protein by Western blot analysis.

CONCLUSIONThe NP9 gene has been successfully isolated and cloned from PBMCs of SLE patients and the corresponding NP9 viral protein expressed in prokaryotic expression vector.

Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Gene Products, env ; biosynthesis ; genetics ; Genetic Vectors ; Humans ; Lupus Erythematosus, Systemic ; genetics ; virology ; Molecular Sequence Data ; Retroviridae ; genetics ; metabolism ; Retroviridae Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of a novel retroviral (NP9) gene transcripts and the possible role of its protein in systemic lupus erythematosus (SLE) patients.

METHODSThe retroviral NP9 gene in SLE patients was isolated and cloned using RT-PCR and TA cloning techniques, and it was analyzed by sequencing. The expression of the NP9 genes in 40 patients with SLE and 48 normal controls using RT-PCR was detected. NCBI BLAST and DNASIS 3.1 software were used to analyze the features of protein of NP9 gene.

RESULTSThe positive ratio (77.5%) of the mRNA expression of the retroviral NP9 gene in SLE patients is significantly higher than that (8.3%) in normal subjects (P<0.01). The recombinant NP9 protein comprises 74 AA with pI 9.59. Amino acid sequence analysis indicates that the retroviral NP9 protein shares higher homologies with several human proteins with important biological functions.

CONCLUSIONSLE patients possess specific novel retroviral NP9 transcripts. The expression of the retroviral NP9 gene may involve in the genesis or development of SLE.

Amino Acid Sequence ; Computational Biology ; Endogenous Retroviruses ; genetics ; metabolism ; Humans ; Lupus Erythematosus, Systemic ; genetics ; physiopathology ; virology ; Molecular Sequence Data ; Retroviridae Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction

The purpose of this review is to confirm the reason resulted the gene silence and explore the countermeasure avoiding the gene silence in transgene plant. The method is to divide the gene silencing into transcriptional gene silencing(TGS) and posttranscriptional gene silencing(PTGS). Several models resulted PTGS were analyzed by RNA threshold model, ectopic pairing and aberrant RNA model and ds-RNA model. The results showed that it was important to decide the phenomena of restraining transgene silencing and the mechanism of PTGS. The strategies of identification of gene function and prevention of virus were presented by RNAi and gene silencing respectively, etc.
Animals ; Gene Silencing ; Models, Genetic ; RNA ; RNA Processing, Post-Transcriptional

OBJECTIVETo observe the clinical efficacy and complications of intradiscal electrothermal therapy for treatment of discogenic low back pain.

METHODSForty patients with discogenic low back pain were treated with intradiscal electrothermal therapy, and the changes in the VAS, functional status and complications after the treatment were analyzed. RESULTS The VAS score was decreased and the functional status improved obviously after the treatment, which caused no severe complications.

CONCLUSIONIntradiscal electrothermal therapy is safe and effective to rapidly achieve pain relief and obviously improve the functional status of patients with discogenic low back pain with few complications.

Adult ; Electric Stimulation Therapy ; Female ; Humans ; Hyperthermia, Induced ; Intervertebral Disc Displacement ; complications ; therapy ; Low Back Pain ; etiology ; therapy ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo transfect pcDNA3.1/hTM plasmids containing human thrombomodulin (hTM) gene into human umbilical vein endothelial cells (HUVECs), and investigate the expression of hTM and anticoagulating function of transfected HUVECs.

METHODSHUVECs were transfected with pcDNA3.1/hTM by lipofectin. Expression of hTM mRNA was determined by semi-quantitative RT-PCR, hTM antigen on HUVECs membrane by immunohistochemistry, and activated protein C (PC) in HUVECs by chronometry. By using a semiautomatic coagulator, the effect of the reacting liquid from transfected HUVECs mixed with PC from normal peripheral blood was assayed.

RESULTSAbout 10% HUVECs were transfected by pcDNA3.1/hTM with high-level hTM mRNA and protein expression. Activated PC produced by pcDNA3.1/hTM group, pcDNA3.1(+)/neo group and untransfected group was (2.80 +/- 0.43) microg/ml, (0.75 +/- 0.08) microg/ml and (0.85 +/- 0.11) microg/ml, respectively. APTT was (51.68 +/- 2.73) s, (38.38 +/- 2.44) s, (39.65 +/- 2.39) s, (33.93 +/- 1.73) s and (34.60 +/- 1.86) s and PT was (21.89 +/- 1.66) s, (20.56 +/- 1.74) s, (20.42 +/- 2.04) s, (19.57 +/- 1.36) s and (20.16 +/- 1.35) s in pcDNA3.1/hTM group, pcDNA3.1(+)/neo group, untransfected group, inactivating PC group and control, respectively.

CONCLUSIONSThe pcDNA3.1/hTM plasmid could be transfected into endothelial cells and expressed biologically functioning hTM protein on HUVECs membrane. Activated PC could inhibit intrinsic coagulation pathway obviously with slight effect on extrinsic pathway.

Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; Gene Expression Regulation ; Genes, Transgenic, Suicide ; Humans ; In Vitro Techniques ; Partial Thromboplastin Time ; Plasmids ; Protein C ; Prothrombin Time ; Thrombomodulin ; genetics ; Transfection ; Umbilical Veins ; cytology

XYL1 gene,which encodes xylose reductase with dual coenzyme activity from Candida tropicalis,was transformed into Pichia pastoris X-33 by expression vector pGAPZB.The recombination strain was immobilized in Ca-alginate beads and fermentation characterization is studied using corn cob hydrolysates.Fermentation conditions were as follow:initial pH value 6.0,30℃,initial cell concentration of 20%,the Liquid volume of 28%,rotation speed 130r/min.The average xylitol yield was 37.5% on the optimum condition.This result is expected to provide a new alternative method for producing xylitol on a large scale by bioconversion.

Objective:To explore the profile and gene functional changes of gut microbiota (GM) in women with postmenopausal osteoporosis (PMOP) in Northwest China, and the correlations between GM and bone mineral density (BMD).Methods:From November 2018 to October 2019, postmenopausal women were screened on their initial visits to our hospital, and 24 new osteoporosis (OP) patients, 30 new osteopenia patients and nine negative controls were recruited. Fecal samples were collected for GM DNA extraction, and Illumina platforms were used for high-throughput sequencing of 16S rRNA and metagenome. Species annotation, GM profile and gene functions were viewed and analyzed.Results:GM profiles were significatly different in different groups, and the LDA scores of Peptostreptococcaceae, Romboutsia, unidentified Clostridiales, Megamonas, Erysipelatoclostridium, Klebsiella and Erysipelatoclostridium ramosum were more than 3 in OP group. Metagenomic sequencing analysis indicated that gene numbers were positively correlated with BMD, and metabolism, carbohydrate metabolism, starch and sucrose metabolism and oxidative phosphorylation were negatively correlated with BMD. Receiver operating characteristic curve(ROC) showed that carbohydrate metabolism, starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism, oxidative phosphorylation, respectively, could identify OP with preferable sensitivity and specificity (areas under curve were 0.70, 0.72, 0.73 and 0.75, respectively). Conclusions:High-throughput sequencing had great potential for GM analysis of postmenopausal women with OP, providing evidence of the correlations between GM and BMD.

[Objective]To investigate the efficacy and safety of ALA-PDT following CO2laser preconditioning for recal-citrant plantar warts.[Methods]Patients with recalcitrant plantar warts were enrolled in this study,and received ALA-PDT treatment following CO2laser preconditioning. Cure rate and side effects were observed.[Results]Twenty patients were en-rolled and 85%(17/20)showed complete clearance of plantar warts after one to three times of ALA-PDT. PDT treatment time was once in two patients(10%),twice in five patients(25%)while three times in 10 patients(50%).No infection or scar tissue was observed.Five(25%)patients were infected with one type of HPV,while 15(75%)patients with two or mul-tiple types of HPV. No difference was observed in complete clearance rate between patients with single or multiple HPV gen-otypes infection.[Conclusions]Superpulse carbon dioxide laser pretreatment enhanced the efficacy of ALA-PDT treatment on recalcitrant plantar warts.Further study is needed to determine the association of HPV genotype with outcome of recalci-trant plantar warts.

OBJECTIVETo explore the relationship between CMV reactivation and KIR haplotype or HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation (HSCT).

METHODSFrom January 2003 to December 2008 the HLA-Cw/KIR genotype of 48 patient-donor pairs were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) and sequence specific nucleotide (PCR-SSOP). Posttransplant CMV reactivation was performed by immune histochemically assay.

RESULTSOf 48 patients, 15 were transplanted from unrelated donors with an antigen mismatch for HLA Cw and 33 patient-donor pairs were matched for HLA-Cw. The CMV reaction rate was 66.7% for HLA-Cw mismatch group and 48.5% for HLA-Cw match group (chi(2) = 1.39, P = 0.2375). Thirty-seven donor-patients pairs belonged to group C1 and 11 to group C2, and CMV reaction rate was 64.9% in group C1 and 18.2% in group C2 (chi(2) = 18.13, P < 0.0001). Twenty-six patients received graft from KIR haplotype A (group A donor) and 22 from KIR haplotype B donors (group B donor) and CMV reaction rate was 57.7% in group A donor and 50.0% in group B donor (chi(2) = 0.28, P = 0.5941). The number of donor activating KIRs (aKIRs) was less than that of recipient aKIRs in 34 patient-donor pairs in which the CMV reaction rate was 70.6%, and the number of donor aKIRs was more than that of recipient aKIRs in 14 patient-donor pairs in which the CMV reactivation was 14.3%. There was a significan difference between the two group (chi(2) = 12.44, P = 0.0004).

CONCLUSIONKIR and HLA-Cw genotypes influence the rate of CMV reactivation following non-T cell deleted unrelated donor hematopoietic cell transplantation.

Genotype ; HLA-C Antigens ; genetics ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; genetics