Objective To study the design and fabricating method of orthosis for patients with leg length discrepancy. Methods Ischial weight bearing orthosis with prosthetic feet was made through efforts from weight,beauty and durabili-ty.The fabrication process included taking a negative plaster cast,modifying the positive model,forming,align-ment,and fitting the device to the patients. Results The orthosis was compensated for the patients'height with good appearance and convenience wearing. Conclusion Ischial weight bearing orthosis with prosthetic feet can help patients reconstruct walking function,therefore can be recommended.

OBJECTIVETo detect the expression of PTEN, PIP3 and cyclin D1 in oral squamous cell carcinoma and precancerous lesions and analyze their correlation.

METHODSImmunohistochemistry SP method was used to detect the expression of PTEN, PIP3 and cyclin D1 in 63 cases of oral squamous cell carcinoma, 29 cases of simple hyperplasia, 33 cases of dysplasia, and 25 cases of normal oral mucosa.

RESULTSThe negative or low expression of PTEN in oral squamous cell carcinoma was 25%, which was remarkably lower than that in other groups. The positive expression of PIP3 in simple hyperplasia, dysplasia and oral squamous cell carcinoma was 66%, 64%, and 76% respectively, which were much higher than those in normal oral mucosa. The positive expression of cyclin D1 in oral squamous cell carcinoma was 49%, which was significantly higher than that in other groups. The negative correlation between PTEN with PIP3, cyclin D1 and the positive correlation between PIP3 and cyclin D1 were observed.

CONCLUSIONSPTEN may play a role in the oncogenesis of oral squamous cell carcinoma, and PTEN may down-regulate the expression of PIP3, and then down-regulate the expression of cyclin D1, which leads to the suppression of cell growth.

Carcinoma, Squamous Cell ; metabolism ; pathology ; Cyclin D1 ; metabolism ; Genes, Tumor Suppressor ; Humans ; Mouth Neoplasms ; metabolism ; pathology ; PTEN Phosphohydrolase ; metabolism ; Precancerous Conditions ; metabolism ; pathology

OBJECTIVETo examine the expression and distribution of NF-kappaBp65, TRAF2, and cyclinD1 and their association with cell apoptosis in oral lichen planus (OLP).

METHODSSixty OLP patients were divided into erosion-atrophy group (n=30) and non-erosion group (n=30) according to their clinical features. Immunohistochemistry with SP method was used to detect the expressions of NF-kappaBp65, TRAF2, cyclinD1 in the 60 OLP and 40 normal oral mucosa (control) specimens. TUNEL assay of randomly selected specimens from 10 normal and 15 OLP cases was performed to detect the cell apoptotic index (AI).

RESULTSCompared with the control group, OLP group showed significantly increased AI of the epithelial cells (67.32-/+18.99) and decreased AI of the lymphocytes (34.12-/+9.89) (P<0.05). In the OLP group, the positivity rates for NF-kappaBp65, TRAF2, and cyclin D1 in the epithelial cells (85.00%, 76.67% and 71.67%, respectively) and in the lymphocytes (91.67%, 86.67% and 70.00%, respectively) were all significantly higher than those in the control group (P<0.05). NF-kappaBp65 expression was significantly increased in the lamina propria in the non-erosion OLP group as compared to the erosion-atrophy group. A positive correlation was noted between lymphocyte NF-kappaBp65 expression and AI of the epithelial cells, but an inverse correlation found between lymphocyte NF-kappaBp65 expression and the AI of the lymphocytes. Lymphocyte TRAF2 and cyclin D1expressions were also inversely correlated to lymphocyte AI. There was a positive correlation between TRAF2 and cyclin D1 expressions and the expression NF-kappaBp65 in the epithelial cells and lymphocytes in OLP.

CONCLUSIONSAccelerated apoptosis of the keratinocytes and inhibition of lymphocyte apoptosis may coexist to contribute to the formation and progression of OLP. NF-kappaBp65 expression, particularly its abnormal nuclear expression, may play a partial role in the pathogenesis of OLP.

Adult ; Aged ; Apoptosis ; Cyclin D1 ; metabolism ; Epithelial Cells ; metabolism ; Female ; Humans ; Lichen Planus, Oral ; metabolism ; Lymphocytes ; metabolism ; Male ; Middle Aged ; TNF Receptor-Associated Factor 2 ; metabolism ; Transcription Factor RelA ; metabolism ; Young Adult

OBJECTIVETo compare clinical curative effects of open surgery (OS) or endovascular repair (EVAR) for patients with abdominal aortic aneurysm (AAA) in China.

DATA SOURCESWe performed a comprehensive search of both English and Chinese literatures involving case studies on retrograde OS or EVAR of AAA in China from January 1976 to December 2010.

STUDY SELECTIONAccording to the inclusion criteria, 76 articles were finally analyzed to compare patient characteristics, clinical success, complications, and prognosis.

RESULTSWe analyzed a total of 2862 patients with 1757 undergoing OS (OS group) and 1105 undergoing EVAR (EVAR group). There was no significant difference in the success rate of the procedures. Operative time, length of ICU stay, fasting time, duration of total postoperative stay, blood loss, and blood transfusion requirements during the procedure were significantly lower in the EVAR group. A 30-day follow up revealed more cardiac, renal, pulmonary, and visceral complications in the OS group (P < 0.01). Low-limb ischemia, however, was more common in the EVAR group (P < 0.05). The 30-day mortality rate, including aorta-related and non-aorta related mortality, was significantly lower in the EVAR group (P < 0.01). In the follow-up period, there were more patients with occlusions of artificial vessel and late endoleak in the EVAR group (P < 0.01). The overall late mortality rate was higher in the OS group (P < 0.01), especially non-aorta-related late mortality and mortality during the fourth to the sixth year (P < 0.01).

CONCLUSIONSEVAR was safer and less invasive for AAA patients. Patients suffered fewer complications and recovered sooner. However, complications such as artificial vessel occlusion, low-limb ischemia, and endoleak were common in EVAR. Clinicians should carry out further research to solve these complications and improve the efficacy of EVAR.

Aged ; Aortic Aneurysm, Abdominal ; surgery ; China ; Endovascular Procedures ; adverse effects ; methods ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Treatment Outcome

OBJECTIVETo construct the programmed cell death 1 ligant 1 (PD-L1) recombination expression vector, express the fusion protein in prokaryotic and analyze the biological action of express product.

METHODSThe whole PD-L1 gene sequence was synthesized after codon optimized. Construct the thioredoxin-(PD-L1) recombination expression vector and express the fusion protein in E. coli. Purified the target protein and analyze the conjugated ability of protein by ELISA.

RESULTSThe PD-L1 recombinant expression vector has been constructed correctly. The target protein has been obtained with which expressed in high efficiency and production. The target protein can conjugate specifically with the PD-1, its specific receptor.

CONCLUSIONWe have obtained the PD-L1 recombinant protein success with high biological activity. The result provide the basic condition for further study on antibody and mutually action between PD-L1 and chronic virus infectious.

Antigens, CD ; chemistry ; genetics ; isolation & purification ; metabolism ; B7-H1 Antigen ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Molecular Weight ; Recombinant Fusion Proteins ; chemistry ; genetics ; isolation & purification ; metabolism

OBJECTIVETo evaluate the association between two single nucleotide polymorphisms located in the promoter of transforming growth factor-β1 receptor 2 (TGFBR2) gene and hypertension in Han Chinese population.

METHODSThe subjects were recruited from the population of cluster sampling survey for essential hypertension (EH) in two townships of Yixing city, Jiangsu province in 2009. Overall, 2012 patients with hypertension and 2116 age (± 2 years) and sex-matched unrelated controls were selected. Epidemiological data, physical measurements results and serum glucose and lipid biomarker were collected and detected. Linkage disequilibrium (LD) analysis were applied and two tagging single nucleotide polymorphisms (tagSNP) in 5' upstream of TGFBR2 gene (rs6785358, -3779A/G; rs764522, -1444C/G) were selected for genotyping and analyzing for the association with hypertension.

RESULTSThe frequencies of AA, AG, GG in case and control of rs6785358 were 1455 (72.3%), 517 (25.7%), 40 (2.0%) and 1582 (74.8%), 490 (23.2%), 43 (2.0%) respectively, and CC, CG, GG of rs764522 were 1524 (75.7%), 464 (23.1%), 24 (1.2%) and 1654 (78.2%), 436 (20.6%), 26 (1.2%) respectively. SNP rs764522 was significantly associated with EH and OR (95%CI) were 1.17 (1.01 - 1.36) (P < 0.05) in dominant model after adjustment for confounding factors such as age, sex, glucose, lipids, smoking and alcohol drinking. Further stratification analysis by age, sex, smoking and alcohol drinking indicated that individuals carrying G allele (CG/GG genotype) of SNP rs764522 had higher susceptibility to EH than CC genotype (OR = 1.21, 95%CI: 1.01 - 1.45) (P < 0.05) in ≥ 55 years group. No statistical significance was detected in the distribution of genotypes and allele frequencies for SNP rs6785358 between cases and controls (P > 0.05). Haplotype analysis showed that no significant frequency difference of haplotype structured by rs6785358 and rs764522 was found between cases and controls (P > 0.05), and no significant blood pressure change was found between genotype variations of rs6785358 and rs764522 (P > 0.05).

CONCLUSIONSNP rs764522 of TGFBR2 gene is associated with increased risk of EH in elderly Han Chinese population.

Aged ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, Transforming Growth Factor beta ; genetics

OBJECTIVETo prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovirus 71.

METHODSTwo peptides, SP55 and SP70, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to establish the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test.

RESULTSTwo high titered anti-VP1 antibodies secreted by the hybridoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1:16 respectively.

CONCLUSIONTwo high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.

Animals ; Antibodies, Monoclonal ; analysis ; immunology ; Antibodies, Viral ; analysis ; immunology ; Capsid Proteins ; analysis ; immunology ; Enterovirus ; chemistry ; immunology ; Enterovirus Infections ; diagnosis ; immunology ; virology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Neutralization Tests

Objective:To investigate the relationship between mitogen-activated protein kinase (MAPK) signaling pathway related signal molecules and the apoptosis of acute promyelocytic leukemia NB4 cells induced by puerariae radix flavones (PRF) and its significance.Methods:The cells were divided into control group [0.025% dimethyl sulfoxide (DMSO) to replace PRF] and 10, 30, 50 μg/ml PRF groups. The proliferation inhibition rate of NB4 cells exposed with PRF for 24, 48 and 72 hours was determined by methyl thiazolyl tetrazolium (MTT) method, and the nuclear morphology was determined by confocal laser scanning microscope after 48 hours. NB4 cells were divided into control group (adding 0.025% DMSO) and 10, 30 and 50 μg/ml PRF with or without 10 μmol/L c-Jun N-terminal kinase (JNK) inhibitor (SP600125) group, and the cells were treated for 48 hours and the changes in the expressions of MAPK pathway related proteins JNK, tumor necrosis factor α (TNF-α), extracellular signal-regulated kinase (ERK) and p38 MAPK were tested by Western blot.Results:10, 30 and 50 μg/ml PRF inhibited the proliferation of NB4 cells in 24, 48 and 72 hours, which was in time- and dose-dependent manners (all P < 0.05). The half-maximal inhibitory concentration (IC 50) at 24, 48 and 72 hours were (40.03±2.23) μg/ml, (22.92±1.72) μg/ml and (17.99±1.48) μg/ml, respectively. The confocal laser scanning microscope showed that NB4 cells displayed distinct apoptotic characteristics after PRF treatment. After co-cultivating NB4 cells with 10 μmol/L SP600125 and different concentrations of PRF for 48 hours, the expression of JNK1 in NB4 cells was suppressed ( P < 0.05), and the expressions of JNK2/3 and p38 MAPK decreased, but the differences were not statistically significant (both P > 0.05). The expressions of ERK1 and ERK2 gradually increased in the single-drug group, while the expression in the combined drug group decreased. The expression of TNF-α in the 50 μg/ml PRF+SP600125 group was down-regulated compared with the 50 μg/ml PRF single-drug group, while the expressions in the 10 and 30 μg/ml PRF+SP600125 groups were up-regulated compared with the 10 and 30 μg/ml PRF single-drug groups. Conclusion:10-50 μg/ml PRF may activate the MAPK signaling pathway through TNF-α. JNK, ERK1/2 and p38 MAPK interact with each other to activate pro-apoptotic related proteins and induce NB4 cells apoptosis.

Objective: To evaluate the outcomes of human leukocyte antigen (HLA) matched unrelated donor hematopoietic stem cell transplantation (MUD-HSCT) for adult acute myeloid leukemia (AML) in a single center. Methods: Consecutive adult AML who received MUD-HSCT in our center from January 2008 to April 2017 were studied retrospectively, comparing with patients undergoing matched sibling donor (MSD) -HSCT in the same period. The rates of overall survival (OS) , disease free survival (DFS) , relapse, non-relapse mortality (NRM) , engraftment, acute and chronic graft-versus-host disease (aGVHD and cGVHD) were analyzed. Results: A total of 247 consecutive cases were enrolled, including 46 patients with MUD-HSCT and 201 with MSD-HSCT. All the patients experienced neutrophil engraftment except for one patient who died early in the MSD group, but the median day of engraftment was longer in the MUD group (15.0 vs 14.0, P=0.017) . The accumulative engraftment rate of platelet was comparable between the two groups (93.5%vs 98.0%, P=0.128) . The accumulative incidences of aGVHD (50.0%vs 46.3%, P=0.421) and cGVHD (37.8%vs 43.0%, P=0.581) were not statistically different between the two groups. Compared with the MSD group, the accumulative NRM rate at+36 months after transplantation was significantly higher in the MUD group (22.0%vs 10.4%, P=0.049) , while the relapse rate was not statistical difference (20.5 vs 28.3%, P=0.189) . Both the 3-year OS (61.6%vs 63.3%, P=0.867) and DFS (57.5%vs 61.6%, P=0.760) were comparable between the two groups. Four independent risk factors were confirmed by the multivariate analysis: patient age ≥45 years old, CR2 or NR before transplantation, a history of extramedullary infiltration and the occurrence of grade Ⅲ-Ⅳ aGVHD. No statistical differences were demonstrated in the survival rate between MUD-and MSD-HSCT in different subgroups. Conclusions: The outcomes, such as GVHD, relapse, OS and DFS, were comparable between MUD-and MSD-HSCT for adult AML, but higher incidence of NRM and longer time to neutrophil engraftment in the MUD group. MUD-HSCT is practical and feasible for adult AML who are lack of MSD.
Graft vs Host Disease ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute/therapy* ; Middle Aged ; Retrospective Studies ; Siblings ; Unrelated Donors

OBJECTIVEObtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity.

METHODSWith the vector of pET-11d, fusion protein was expressed in Escherichia coli BL21 (DE3) after induced by IPTG. The protein was then purified by DEAE negative ion exchange chromatography and Ni2+ affinity chromatography. Western Blot analysis was used to detect the antigenicity of the fusion protein. At the same time, the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA.

RESULTSHigh purified hepatitis C virus subunit fusion protein was obtained successfully. The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers.

CONCLUSIONThe hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity. It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.

Animals ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Hepacivirus ; genetics ; metabolism ; Immunoassay ; Rabbits ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Viral Proteins ; genetics ; immunology ; metabolism