BACKGROUND:Stress shielding in the Achil es tendons induces over-expression of tumor necrosis factor-α. The degree of tendon contracture remains unclear after the intervention with tumor necrosis factor inhibitor.
OBJECTIVE:To investigate the effects of tumor necrosis factor-αon tendon contracture and the preventive effects of tumor necrosis factor inhibitor (etanercept) on tendon contracture by observing the morphological changes of the stress-shielded Achil es tendons after the intervention with etanercept.
METHODS:A total of 20 healthy male Sprague-Dawley rats were randomly divided into experimental and model groups after stress shielding in Achil es tendons of rat left hind limb. Five rats from either group were randomly selected, and their right hind limbs were considered as normal controls. Immediately after model induction, the rats in the experimental group were subjected with 0.6 mg/kg etanercept, and those in the model group were subcutaneously treated with 1 mL phosphate buffered saline. According to half-life of etanercept, the two groups were separately injected three times. At 2 weeks after intervention, the morphological changes of the Achil es tendons were observed using gross examination and transmission electron microscope.
RESULTS AND CONCLUSION:On gross examination, the Achil es tendons in the experimental group were significantly smoother and smal er than those of the model groups, but thicker than those of the normal control group. Under a transmission electron microscope, the col agen fibrils of the model group were looser and more disordered than those of the experimental group. The col agen fibrils of the experimental group were similar to those of the normal control group in cross section and longitudinal section. These indicated that tumor necrosis factor-αantagonist can obviously prevent stress shielding-induced tendon contracture at 2 weeks.

Nowadays, proteomics focuses on quantitative analysis rather than qualitative. In the field of quantitative proteomics, Isobaric tags for relative and absolute quantitation (iTRAQ) is one of the most widely used techniques. The advantage of iTRAQ is high throughput, high stability and free of the restriction of sample property. iTRAQ is suitable for almost all kinds of samples, and up to 8 samples can be analyzed simultaneously by commercially available kit. Along with the development of techniques, more and more mass spectrometry (MS) platforms are used in iTRAQ experiments and the accuracy of iTRAQ has been improved. iTRAQ has been applied to studies of microorganism, animal, plant, medical and protein post-translational modification. Here we review the recent progress in the development of iTRAQ and its applications in quantitative proteomics.
Animals ; Humans ; Mass Spectrometry ; Proteomics ; methods

Adolescent ; Adult ; Aged ; Carcinoma ; metabolism ; pathology ; Core Binding Factor Alpha 3 Subunit ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Protein Array Analysis ; methods ; Young Adult

Objective To observe the relationship between early lactate clearance and APACHEⅡ in critically ill patients. Methods In 121 critically ill patients,APACHE Ⅱ and lactate clearance rate were recorded. According to APACHE Ⅱ score,all patients were divided into≤10 score group, (11 ～20) score group,(21 ～30) score group and ＞ 30 score group, then compared the level of the early lactate clearance rate. The early lactate clearance rate were also compared between survival group and death group. Then the relationship between early lactate clearance and APACHE Ⅱ were analyzed. Results In( 11 ～20) score group,the early lactate clearance rate was lower than those in ≤ 10 score group, but the difference was not significant ( P ＞ 0. 05 ). The early lactate clearance rate in (21 ～ 30) score group ( 18. 35 ± 10. 01 ) % was lower than those in ( 11 ～ 20) score group (27.35 ± 10. 22) % ( t = 3.481, P ＜ 0. 01 ),in ＞ 30 score group( 11.98 ± 9. 93 )% those was lower compared with (21 ～30) score group( t = 2. 968, P ＜ 0. 01 ). In death group, APACHE Ⅱ score(28. 1 ± 6. 7 ) was higher than that in survival group ( 18. 8 ± 8. 4) ( t = 3. 030, P ＜0. 01 ), the early lactate clearance rate was lower ( t = 3. 619, P ＜ 0. 01 ). APACHE Ⅱ score correlated well with the mean level of the early lactate clearance rate ( r = - 0. 641, P ＜ 0. 01 ). Conclusion The lactate clearance rate was the good fator on evaluation of condition and prognosis in the critically ill patients.

OBJECTIVE:To investigate clinical significance of programmed cell death 5 (PDCD5) protein in serum of pa-tients with lung cancer. METHODS:80 lung cancer inpatients were selected from the First Affiliated Hospital of Shihezi University School of Medicine(hereinafter referred to asour hospital)as lung cancer group;60 healthy volunteers were selected from our hospital at the same period as normal group. ELISA was used to test the expression of PDCD5 protein,and the relationship of PDCD5 protein with clinical pathological features of lung cancer patients were analyzed. RESULTS:The expression of PDCD5 pro-tein in normal group was significantly higher than lung cancer group,with statistical significance (P<0.05). The expression of PDCD5 protein in lung caner patients was not associated with gender,smoking history and pathological type(P>0.05);it was de-creased as the decrease of tumor differentiation degree,with statistical significance(P<0.05). The expression of PDCD5 protein in patients with carcinoembryonic antigen(CEA)<5.6μmol/L was significantly higher than those with CEA≥5.6μmol/L;the expres-sion of PDCD5 protein in patients with cytokeratin 19 soluble fragment (CYFRA21-1)<5.6 μmol/L was significantly higher than those with CYFRA21-1≥5.6 μmol/L,with statistical significance(P<0.05). The expression of PDCD5 protein in patients with Ⅰ-Ⅱ stage lung cancer was significantly higher than Ⅲ-Ⅳ stage lung cancer patients;the expression of PDCD5 protein in patients without distant metastasis was significantly higher than those with distant metastasis. The expression of PDCD5 protein was in de-creasing as the increase of the number of metastasis site,with statistical significance(P<0.05). CONCLUSIONS:PDCD5 protein in serum of patients with lung cancer shows low expression level,which is related to tumor differentiation degree,tumor marker level as CEA and CYFRA21-1,tumor stage and distant metastasis,etc. The detection of PDCD5 protein may contribute to clinical diagnosis of lung cancer.

Objective The generation of drug resistance often leads to the failure of the bladder cancer chemotherapy.P-glycoprotein (P-gp) is an ATP-dependent drug efflux pump linked to development of multidrug resistance in cancer cells.The laboratory has successfully established adriamycin-resistant human bladder cancer cell line (pumc-91/ADM) from its parental cell line (pumc-91).According to the drug resistant spectrum analysis,pumc-91/ADM cell line exhibited the characteristics of multi-drug resistance.However,the expression of P-gp in two cell lines was still unknown.In this paper,there was a comparison between pumc-91/ADM and pumc-91 about the differential expression of P-gp.Methods To determine the expression and location of P-gp in pumc-91 and pumc-91/ADM,qRT-PCR,Western blot and immunocytochemistry were applied in the experiment.qRT-PCR was implemented to research the expression of P-gp mRNA in two cell lines (pumc-91/ADM and pumc-91).Western blot was adopted to investigate the expression of P-gp protein in pumc-91 and pumc-91/ADM cell lines.Immunocytochemistry technique was used to explore the cellular location of P-gp and affirm its expression in two cell lines visually.Student's t-test was employed for statistical analysis and P ＜ 0.05 was considered statistically significant.Results qRT-PCR analysis revealed that the expression of P-gp mRNA was upregulated in drug-resistant cell line pumc-91/ADM compared to parental cell line pumc-91.To normalize for differences in the amount of total RNA,GAPDH was selected as an endogenous RNA control.Compared with pumc-91,the expression of P-gp mRNA was upregulated 7.74 fold in pumc-91/ADM (t =11.97,P ＜ 0.05).Consistent with the qRT-PCR result,Western blot confirmed the protein of P-gp expressed differentially in two cell lines.The expression of P-gp protein was significantly increased in pumc-91/ADM compared to pumc-91.According to the results,the differences between pumc-91 and pumc-91/ADM had statistical significance (t =4.35,P＜0.05).Immunocytochemical analysis results demonstrated that P-gp was not only located in cell membrane but also in cytoplasm of the two cell lines.The expression of P-gp in pumc-91/ADM increased distinctly.The difference was statistically significant (t =11.41,P ＜ 0.05).Conclusion Compared with pumc-91,the expression of P-gp in pumc-91/ADM was significantly upregulated.

Behavior Therapy ; Chronic Disease ; Cognitive Therapy ; Humans ; Pain ; psychology ; Pain Management

AIM: To investigate the differences in the distribution of SRY-related HMG box 5 (SOX5) gene single nucleotide polymorphisms (SNPs) among stable chronic obstructive pulmonary disease (COPD) patients, COPD with pulmonary hypertension (PH) patients and healthy controls, and to explore the association of the SOX5 SNPs in COPD-related PH.METHODS: From April 2013 to April 2015, 250 patients with stable COPD were enrolled continuous-ly in Ningxia People’s Hospital according to COPD treatment guidelines (2013 edition).All the patients received echocar-diography, and were divided into COPD with PH group [pulmonary artery systolic pressure (PASP)≥50 mmHg, n =103] and COPD without PH group (PASP <50 mmHg, n =147).The healthy persons (matched for age, sex, race and smoking index, n =127) were selected as control group at the same period.Genotyping of SOX5 gene rs10842262 and rs11046966
loci was performed using MassARRAY genotyping system ( Sequenom).Genotype frequencies were calculated.RE-SULTS: Age, sex and smoking index showed no significantly difference between control group and COPD group, neither between COPD with PH group and COPD without PH group.Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci between control group and COPD group was of significant difference (P<0.05).Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci showed no significant difference between COPD with PH group and COPD without PH group.CONCLUSION: SOX5 gene rs10842262 and rs11046966 loci may play an important role in COPD, but not in COPD-related PH.

BACKGROUNDArthritogenic T lymphocytes with common T cell receptor (TCR) Vbeta clonotypes, infiltrating in the articulars of rheumatoid arthritis (RA) patients, play a central role in the pathogenesis of RA. TCR Vbeta5.2 and TCR Vbeta8.2 are the main pathogenic T cell clonotypes in the course of collagen-induced arthritis (CIA) progression in Lewis rats. To investigate a TCR-based immunotherapy for RA, we constructed recombinant DNA vaccines encoding TCR Vbeta5.2 and TCR Vbeta8.2, and evaluated the inhibitive effects of the two vaccines on CIA rats.

METHODSGenes encoding TCR Vbeta5.2 and TCR Vbeta8.2 were amplified by RT-PCR from spleen lymphocytes of Lewis rats and cloned into the eukaryotic expression vector pTargeT. The expression of vaccines was confirmed by RT-PCR and immunohistochemistry. The inhibitive effects of the vaccines on articulars of CIA rats were assessed with arthritis index evaluation and histology. Interferon gamma (IFN-gamma) and interleukin (IL)-4 production by spleen lymphocytes were tested with enzyme-linked immunospot assay (ELISPOT) technique, the changes in peripheral CD4(+) and CD8(+) lymphocyte populations were tested by flow cytometry, and the level of anti-CII antibody in serum was assayed by enzyme-linked immunosorbent assay (ELISA).

RESULTSRecombinant DNA vaccines pTargeT-TCR Vbeta5.2 and pTargeT-pTCR Vbeta8.2 were successfully constructed. Both vaccines inhibited CIA, which alleviated the arthritis index score (P < 0.05), decreased the level of IFN-gamma (P < 0.05), and reduced the ratio of CD4(+)/CD8(+) lymphocytes (P < 0.05) and the anti-CII antibody in serum (P < 0.05). In addition, the histological change in DNA-vaccinated rats was less serious than CIA rats. Compared to pTCR Vbeta 8.2 and pTCR Vbeta 5.2 groups, the group that was injected with a combination of the two vaccines showed stronger inhibitive effects on CIA than either individual vaccine.

CONCLUSIONThe recombinant plasmids pTargeT-TCR Vbeta5.2 and pTargeT-TCR Vbeta8.2 have obvious inhibatory effects on CIA rats and better effects could be achieved when the vaccines were used in combination.

Animals ; Arthritis, Experimental ; metabolism ; prevention & control ; CD4-Positive T-Lymphocytes ; drug effects ; CD8-Positive T-Lymphocytes ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Muscles ; drug effects ; metabolism ; Peptide Fragments ; antagonists & inhibitors ; Rats ; Rats, Inbred Lew ; Receptors, Antigen, T-Cell, alpha-beta ; antagonists & inhibitors ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; pharmacology

Adenocarcinoma, Follicular ; pathology ; physiopathology ; Female ; Humans ; Kidney Diseases ; pathology ; physiopathology ; Kidney Neoplasms ; physiopathology ; Thyroid Neoplasms ; pathology ; physiopathology ; Young Adult