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1.
Article in Chinese | WPRIM | ID: wpr-888326

ABSTRACT

OBJECTIVE@#To establish a risk assessment and prediction system for early osteonecrosis of the femoral head (ONFH) in order to predict the collapse risk.@*METHODS@#The risk assessment system for early necrosis and collapse of femoral head was established based on the combination of Steinberg stage, ABC typing and the proportion of the proximal sclerotic rim. Firstly, Steinberg stage system was applied. ABC typing was applied to predict risk in stage I, type C was risk free, type B was low risk, type A and type BC were medium risk, type A-C and type AB were high risk. The classification of proximal sclerotic rim was first applied when the Steinberg stage was Ⅱ-Ⅲ, and type 2 was expected to be low risk. If the classification of proximal sclerotic rimwas type 1, then the ABC typing was applied, type C was risk-free, type B was low risk, type A and type BC were medium risk, type A-C and type AB were high risk. According to this prediction system, the collapse risk of femoral head in 188 cases(301 hips) were predicted by retrospective analysis. All the hips were enrolled at the out-patient department of orthopedic in Guang'anmen Hospital attached to China Academy of Chinese Medical Science. The consistency of the prediction results of three doctors and one doctor at different times were evaluated.@*RESULTS@#Among them, 136 cases were male, 52 were female. 75 cases were single hip, 113 were double hip. The age of the patients wa 19 to 64(42.61±12.07) years. The natural course of disease was 0.33 to 5.00(3.62±1.93) years. 206 hips in 301 hips had collapsed, with a collapse rate of 68.44%. In the risk-free group, none hip had collapsed, with a collapse rate of 0%. In the low-risk group, 9 hip in 91 hips had collapsed, with a collapse rate of 9.89%. In the medium-risk group, 12 hip in 19 hips had collapsed, with a collapse rate of 63.16%. And in the high risk group, 185 hips in 190 hips had collapsed, with a collapse rate of 97.37%. They were significantly differences in their collapse rate (@*CONCLUSION@#The risk assessment and prediction system for early ONFH selects different methods to predict the risk of collapse according to the imaging characteristics of different stages, which is combines with the comprehensive assessment of multiple risk factors. The system is applicable to a wide range, simple operation and convenient for clinical application.


Subject(s)
Adult , China , Female , Femur Head/diagnostic imaging , Femur Head Necrosis/epidemiology , Humans , Male , Middle Aged , Retrospective Studies , Risk Assessment , Treatment Outcome , Young Adult
2.
Article in Chinese | WPRIM | ID: wpr-772583

ABSTRACT

OBJECTIVE@#To explore the influencing factors and possible mechanism of axial symptoms(AS) after C₃-C₇ single open-door laminoplasty in patients with chronic compression cervical myelopathy.@*METHODS@#The clinical data of 32 patients with multi-segment chronic compression cervical cord disease treated by C₃-C₇ single open-door laminectomy from May 2012 to July 2016 were retrospectively analyzed. Including cervical spondylotic myelopathy of 14 cases, developmental cervical stenosis complicated with cervical myelopathy of 8 cases, ossification of posterior longitudinal ligament(OPLL) of 10 cases. There were 17 males and 15 females, aged from 47 to 82 years old with an average of 57.46 year, the course of disease was 5 to 35 months with an average of 22.4 months. The opening angle(OA), cervical curvature angle(CA), preoperative spinal cord compression rate(PSCR) and postoperative spinal cord shift (PSCS) were recorded. After 2 weeks of surgery, determining whether occurred an AS condition according to the AS assessment criteria, the patients were divided into a axial symptom group and a non-axial symptom group, the general data and imaging parameters of the two groups were compared and the factors that may be postoperative AS were analyzed by binary Logistic regression analysis.@*RESULTS@#At 2 weeks after operation, 13 patients occurred AS. There was no significant difference in gender, age and course of disease between axial symptom group and a non-axial symptom group (>0.05). In axial symptom group, OA was(36.76±9.35)°, CA was(11.53±4.36)°, PSCR was(27.83±1.72)%, PSCS was (3.17±0.81) mm, while in non-axial symptom group, above items were (33.03±10.52)°, (7.71±4.73)°, (25.16±3.59)%, (2.43±0.95) mm, respectively, there was significant difference in CA, PSCR, PSCS between two groups(0.05). The results of the binary Logistic regression analysis of 3 parameters(OA, PSCR, PSCS) and AS showed OA and PSCR were eliminated in dependent variables, and the partial regression coefficient of PSCR was 0.311, and =0.031.@*CONCLUSIONS@#CA, PSCR, and PSCS are related influencing factors of AS, and PSCS is a high risk factor for AS. C₄,₅ nerve traction caused by posterior spinal movement, postoperative dural self-expansion causes greater traction of the spinal cord, excessive deformation of the cervical spinal cord causes autonomic nerve damage or necrosis that dominates blood vessels may be the pathogenesis of AS, but this is only a theoretical inference, and further improved experiment is necessary to verify it in the future.


Subject(s)
Aged , Aged, 80 and over , Cervical Vertebrae , Female , Humans , Laminectomy , Laminoplasty , Male , Middle Aged , Retrospective Studies , Spinal Cord Diseases , Treatment Outcome
3.
Article in Chinese | WPRIM | ID: wpr-776165

ABSTRACT

OBJECTIVE@#To observe the open angle (OA), cervical curvature angle (CA), preoperative spinal cord compression rate(PSCR), postoperative spinal cord shift (PSCS) in patients with chronic compressive cervical myelopathy undergoing C3-7 single open laminoplasty, and to explore the possible mechanism and influencing factors of postoperative average spinal cord drift, so as to provide objective basis for predicting PSCS.@*METHODS@#From May 2012 to July 2016, 32 patients with multi-segmental chronic compressive cervical myelopathy who underwent single-door laminoplasty in our department were analyzed retrospectively, including 14 cases of cervical spondylotic myelopathy, 8 cases of developmental cervical spinal stenosis with cervical myelopathy, and 10 cases of ossification of posterior longitudinal ligament. The OA of cervical spine was measured on CT, the CA was measured on X-ray, the PSCR and PSCS were measured on MRI. The patients were divided into two groups according to PSCS(group A>=2.5 mm, group B0.1), and the partial regression coefficients of OA and PSCR were 0.113 and 0.059 respectively.@*CONCLUSIONS@#PSCS is the result of OA, CA and PSCR, among which PSCR has the most important influence, OA is the second, CA is the least. PSCS can be predicted by 0.059×OA+0.113×PSCR-2.266 equation, which provides a theoretical basis for preoperative evaluation of spinal cord decompression after surgery.


Subject(s)
Aged , Cervical Vertebrae , Female , Humans , Laminectomy , Laminoplasty , Male , Middle Aged , Retrospective Studies , Spinal Cord Diseases , Treatment Outcome
4.
Article in Chinese | WPRIM | ID: wpr-256517

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of claudin-3 in colorectal carcinoma and its association with the occurrence, progression and prognosis of colorectal cancer.</p><p><b>METHODS</b>Forty surgical specimens of colorectal carcinoma and 22 adjacent normal tissues resected between October, 2010 and January, 2013 at Nanfang Hospital were examined for claudin-3 expression using immunohistochemistry, which was analyzed in association with the clinicopathological parameters and the survival of the patients.</p><p><b>RESULTS</b>Claudin-3 was expressed mainly on the cell membrane, and its positivity rate was significantly higher in cancer tissues than in normal tissues (92.50% vs 59.09%, P<0.05). In 13 cases claudin-3 expression was detected in both the cancer tissues and adjacent normal tissues with average expression scores of 4.538 and 3.269, respectively (P<0.05). In the cancer tissues, the strongly positive expression rate was significantly higher in poorly differentiated tissues (85.71%) than in well (21.43%) and moderately (36.48%) differentiated tissues (P<0.05), and was higher in cases with lymph node metastasis than in those without (61.11% vs 22.72%, P<0.05). The strongly positive expression rate of claudin-3 was not correlated with the patients'age, gender, tumor location or tumor size (P>0.05). Of the 33 cancer patients followed up, 14 had a postoperative survival time no longer than 3 years and 19 had longer survival time, and their average expression scores differed significantly (4.50 vs 3.526, P<0.05).</p><p><b>CONCLUSION</b>Claudin-3 is over-expressed in colorectal cancer tissues, and its high expression may promote the occurrence and progression of colorectal cancer. Claudin-3 may serve as a molecular biomarker for early diagnosis and prognostic evaluation.</p>

5.
Article in Chinese | WPRIM | ID: wpr-275122

ABSTRACT

PLA-α-asarone nanoparticles were prepared by using organic solvent evaporation method, and their in vivo distribution and brain targeting after intranasal administration were studied as compared with intravenous administration. The results showed that brain targeting coefficient of PLA-α-asarone nanoparticles after intranasal and intravenous administration was 1.65 and 1.16 respectively. The absolute bioavailability, brain-targeting efficiency and the percentage of nasal-brain delivery of PLA-α-asarone nanoparticles were 74.2%, 142.24 and 29.83%, respectively after intranasal administration. The results of fluorescence labeling showed that the fluorescent intensity of coumarin-6 in the brain tissue was the highest after intranasal administration of PLA-α-asarone fluorescent nanoparticles, achieving the purpose of brain-targeted drug delivery. The fluorescent intensity of coumarin-6 in liver tissue after intravenous administration of PLA-α-asarone nanoparticles was much higher than that after intranasal administration, indicating that intranasal administration of PLA-α-asarone nanoparticles could decrease drug-induced hepatotoxicity. In addition, the fluorescent intensity of coumarin-6 in lung tissue was weaker after intranasal administration, which solved the shortcomings of intranasal administration of α-asarone dry powder prepared by airflow pulverization method. In vivo studies indicated that PLA-α-asarone nanoparticles after intranasal administration had a stronger brain targeting as compared with intravenous administration.

6.
Article in Chinese | WPRIM | ID: wpr-328246

ABSTRACT

<p><b>OBJECTIVE</b>To observe the distribution of constitution types of Chinese medicine (CM) in patients with osteonecrosis of femoral head (ONFH).</p><p><b>METHODS</b>Totally 130 ONFH patients were recruited. Constitution types of CM were identified in all patients. Distribution features of constitution types of CM in ONFH patients were observed. The differences of distribution in gender, age, single or bilateral hips, course of disease, staging, cause, and region were also analyzed.</p><p><b>RESULTS</b>Seventy patients were of complicated constitutions, while 60 patients were of single constitution. Among the 60 single constitution cases, yang-deficiency constitution [18 (13.9%)], damp-heat constitution [10 (7.7%)], blood-stasis constitution [7 (5.4%)], and qi-deficiency constitution [7 (5.4%)] were mainly distributed. Of the complicated constitutions, yang-deficiency dominated constitution occupied the top ratio [30 (23.1%)], followed by blood-stasis dominated constitution [15 (11.5%)], damp-heat dominated constitution [9 (6.9%)]. By putting them together, yang-deficiency constitution occupied the top constitution of CM [48 (36.9%)], followed by blood-stasis constitution [ 22 (16.9%)] and damp-heat constitution [19 (14.6%)]. The aforesaid three constitutions accounted for 68.5% of the total. There were no statistical distribution differences in gender, age, single or bilateral hips, course of disease, staging, or cause.</p><p><b>CONCLUSION</b>Yang-deficiency constitution, damp-heat constitution, and blood-stasis constitution were liable constitutions of CM in ONFH patients.</p>


Subject(s)
Femur , Pathology , Humans , Medicine, Chinese Traditional , Osteonecrosis , Drug Therapy , Yang Deficiency
7.
Chinese Journal of Virology ; (6): 285-291, 2014.
Article in Chinese | WPRIM | ID: wpr-356602

ABSTRACT

Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.


Subject(s)
Animals , Bombyx , Virology , Gene Deletion , Gene Expression Regulation, Viral , Nucleopolyhedroviruses , Genetics , Physiology , Transcription, Genetic , Viral Proteins , Genetics , Metabolism , Virus Replication
8.
Chinese Medical Journal ; (24): 274-279, 2013.
Article in English | WPRIM | ID: wpr-331281

ABSTRACT

<p><b>BACKGROUND</b>Serum tumor markers have always been of clinical importance in the diagnosis, monitoring disease progression and therapy efficacy for patients with malignant diseases. However, elevated serum tumor markers are found in some benign conditions, especially in chronic kidney disease (CKD). The elevation of them in CKD might cause confusion and misuse of these tumor markers. We conducted this retrospective study to investigate which of the five widely used tumor markers including carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), cytokeratin 19 fragment antigen 21-1 (Cyfra21-1), squamous cell carcinoma antigen (SCC) and neuron specific enolase (NSE) are affected markedly by CKD, in order to use them more effectively.</p><p><b>METHODS</b>Serum tumor marker concentrations, biochemical, hematological parameters, and urinalysis were measured in CKD patients and healthy controls. The positive rate and median tumor markers' level in CKD patients and controls, and those in CKD patients stratified by CKD grade were compared using nonparametric rank tests. Correlation analysis of serum tumor markers and other parameters in CKD patients were performed using the Spearman correlation coefficient. Multivariate Logistic regression analysis was used to estimate the important variables that caused elevated serum concentrations of these markers in CKD patients.</p><p><b>RESULTS</b>The overall positive rates and serum concentrations of Cyfra21-1, SCC, CEA in CKD group were significantly higher than those in control group. Positive rate and serum concentrations of those tumor markers increased as kidney function decreased. Both univariate analysis and multivariate regression analysis showed that the elevations of those tumor markers were not only associated with kidney function, but also with nutritional status.</p><p><b>CONCLUSIONS</b>Serum concentrations of Cyfra21-1, SCC, CEA are significantly influenced by kidney function, as well as nutritional status. Therefore, in clinical work, the indices of kidney function and nutritional status could be simultaneously measured to improve interpretation of the results of those tumor marker concentrations.</p>


Subject(s)
Adult , Aged , Antigens, Neoplasm , Blood , Biomarkers, Tumor , Blood , Carcinoembryonic Antigen , Blood , Female , Glomerular Filtration Rate , Humans , Keratin-19 , Blood , Logistic Models , Male , Middle Aged , Nutritional Status , Renal Insufficiency, Chronic , Blood , Retrospective Studies , Serpins , Blood , alpha-Fetoproteins
9.
Article in Chinese | WPRIM | ID: wpr-346490

ABSTRACT

To observe the clinical effect of Jianpi Bushen formula to prevent the collapse of osteonecrosis of femoral head (ONFH) of type phlegm and blood stasis obstructing the collaterals. 50 cases (including 73 hips) of non-collapse ONFH (ARCO I, II, III a) were selected from the out-patient department of orthopedic in Guang'anmen Hospital attached to China Academy of Chinese Medical Science. All the cases fit for diagnostic criteria were given Jianpi Bushen formula and followed up. Staging criteria was ARCO classification. Harris score evaluated the hip function. The mean follow-up time was 4.2 years (3-5.4 years). After a mean of 4.2 years (3-5.4 years) followed-up, 12 hips collapse occurred while 61 hips not (the non-collapse rate was 83.56%). Collapse did not occurred among the all 7 hips at stage ARCO I (the non-collapse rate was 100%). Among the 49 hips at stage ARCO II, 9 hips collapse occurred while 40 hips not (the non-collapse rate was 81.63%). Among the 17 hips at stage ARCO III a, 3 hips collapse occurred while 14 hips not (the non-collapse rate was 82.35%). Kaplan-Meier analysis showed the average survival time of non-collapse was 5 years (4. 8-5.2 years). The 3 year survival rate of non-collapse was 92.5%. The 4 year survival rate was 74% and the condition tended to be stability trend. The Harris score was 71.93 +/- 11.25 before treatment and 81.63 +/- 12.16 after treatment, significantly different. These results suggest that: Jianpi Bushen formula is an effective method for treating ONFH of type phlegm and blood stasis obstructing the collaterals. It can delay or prevent the collapse of femoral head and significantly improve the hip function.


Subject(s)
Adult , Aged , Drugs, Chinese Herbal , Female , Femur Head Necrosis , Drug Therapy , Follow-Up Studies , Humans , Male , Middle Aged , Treatment Outcome , Young Adult
10.
Article in Chinese | WPRIM | ID: wpr-332737

ABSTRACT

This study was aimed to investigate the expression level and mechanism of microRNA-223 and LMO2 in acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) cells and the mechanism. MicroRNA-223 mimics was transfected to increase the expression of MicroRNA-223 in the lymphocytes sorted by ficoll separation from the bone marrow mononuclear cells (BMMNC) of ALL and CLL patients. MicroRNA-223 inhibitor was transfected to decrease the expression of the MicroRNA-223 in the lymphocytes of normal controls. Then the expression of the MicroRNA-223 and LMO2 in transfected lymphocytes before and after cultivating for 72 hours were detected by RT-PCR, the apoptosis and cell cycle of these cells were measured by flow cytometery. The results indicated that before the transfection, the expression of MicroRNA-223 in ALL and CLL cells was (433.11 ± 144.88), which was significantly lower than that in the normal lymphocyte (949.59 ± 267.39); the expression of LMO2 was (807.10 ± 238.41), which was significantly higher than that in the normal lymphocytes (455.32 ± 176.83) (P < 0.05); after the transfection, the expression of MicroRNA-223 was (571.86 ± 142.00) in ALL and CLL cells, which was significantly higher than that before transfection (P < 0.05), but the expression of LMO2 was significantly lower than that before transfection (651.97 ± 230.12) (P < 0.05); in the normal control the expression of MicroRNA-223 obviously decreased (646.32 ± 172.93) (P < 0.05), the expression of LMO2 was significantly increased (541.27 ± 158.86.2) (P < 0.05). After transfection, the cell cycle G1/G2 phase and apoptosis changed in ALL and CLL cells. Before transfection the cell ratio in cell cycle G1/G2 phase was (94.75 ± 3.15)%, the cell ratio in S phase was (5.14 ± 3.12)%; after transfection the cell ratio in cell cycle G1/G2 phase was (97.03 ± 2.08)% and obviously increased (P < 0.05), the cell ratio in S phase was (2.97 ± 2.08)% and significantly decreased (P < 0.05). Before transfection the apoptosis rate was (54.47 ± 8.72)%, and obviously was higher than that after transfection (60.48 ± 8.81)%. And in the normal control, the cell ratio in G1/G2 phase was significantly higher than that after transfection [(96.73 ± 2.26)%, (94.55 ± 2.77)%, P < 0.05)], and the cell ratio in S phase was significantly increased [(3.25 ± 2.26)%, (5.45 ± 2.77)% (P < 0.05)]. The apoptotic rate in the ALL and CLL patients was significantly higher than that after the transfection [(54.47 ± 8.72)% vs (60.48 ± 8.81)%, respectively (P < 0.05)]. The apoptotic rate in the normal control was significantly lower than that after the transfection [(59.02 ± 10.20)%, (51.96 ± 10.20)%, respectively (P < 0.05)]. It is concluded that the expression of MicroRNA-223 decreases, and the expression of LMO2 increases in lymphocytic leukemia cells which leads to the lymphocytes over-proliferation and abnormal apoptosis, thus may be one of pathogenesis in lymphocytic leukemia.


Subject(s)
Adaptor Proteins, Signal Transducing , Genetics , Metabolism , Adolescent , Adult , Aged , Apoptosis , Case-Control Studies , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Female , Humans , LIM Domain Proteins , Genetics , Metabolism , Leukemia, Lymphocytic, Chronic, B-Cell , Genetics , Metabolism , Male , MicroRNAs , Genetics , Metabolism , Middle Aged , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Metabolism , Proto-Oncogene Proteins , Genetics , Metabolism , Transfection , Young Adult
11.
Chinese Journal of Stomatology ; (12): 669-671, 2013.
Article in Chinese | WPRIM | ID: wpr-274161

ABSTRACT

<p><b>OBJECTIVE</b>To assess the effect of removable partial dentures (PRD) restoration on oral health-related quality of life (OHRQOL) of subjects with shortened dental arches (SDA) using the Chinese version of the Oral Health Impact Profile (OHIP-14).</p><p><b>METHODS</b>Consecutive patients with shortened dental arches were recruited from the Department of Prosthodontics, Foshan Chancheng Hospital of Stomatology. The Chinese version of OHIP-14 was administered to each subject before treatment and after treatment. The subjective outcomes of removable partial dentures therapy on SDA were collected and the pre- and post-treatment scores of the Chinese version of OHIP-14 were compared.</p><p><b>RESULTS</b>After treatment, significant decrease in patients' total score of OHRQOL [before: 9 (6, 12) ; after: 4 (2.25, 6)] was detected, as well as physical pain subscale, physical disability subscale and handicap subscale; and the score of OHRQOL in function limitation subscale increased [before: 0(0,0); after: 2(1, 2)].</p><p><b>CONCLUSIONS</b>The main impacts of shortened dental arch with intact anterior region affecting patients' OHRQOL are physical disability.From a quality-of-life perspective, patients with SDA can perceive benefits from RPD.</p>


Subject(s)
Adolescent , Adult , Dental Arch , Pathology , Denture, Partial, Removable , Psychology , Humans , Jaw, Edentulous, Partially , Pathology , Psychology , Oral Health , Quality of Life , Sickness Impact Profile , Surveys and Questionnaires , Young Adult
12.
Article in English | WPRIM | ID: wpr-299415

ABSTRACT

<p><b>OBJECTIVE</b>To observe the influence of Shenqing Recipe (SQR), a kind of Traditional Chinese Medicine, on the morphology and quantity of colonic interstitial cells of Cajal (ICC) in trinitrobenzene sulfonic acid (TNBS)-induced rat colitis, and to investigate the possible mechanism of SQR in regulating intestinal dynamics.</p><p><b>METHODS</b>Sixty rats were randomly divided into normal control, model 1, model 2, mesalazine, and high-dose, and low-dose SQR groups with 10 rats in each group. TNBS (10 mg) dissolved in 50% ethanol was instilled into the lumen of the rat colon of the latter five groups to induce colitis. On the 4th day after administration of TNBS, each treatment group was administered one of the following formulations by enteroclysis gavage once a day for 7 days: 600 mg•kg⁻¹•d⁻¹ mesalazine, 2.4 g•kg⁻¹•d⁻¹ SQR, and 1.2 g•kg⁻¹•d⁻¹ SQR. Model 2 rats received normal saline solution. After 7 days colonic samples were collected. While the colonic samples of model 1 group were collected on the 3rd day after TNBS administered. Ultrastructure of ICC in the damaged colonic tissues was observed with transmission electron microscope. Expression of c-kit protein in colonic tissue was determined by immunohistochemical staining and Western blot.</p><p><b>RESULTS</b>The ultrastructure of colonic ICC in the rat model of TNBS-induced colitis showed a severe injury, and administration of SQR or mesalazine reduced the severity of injury. Similarly, the expression of c-kit protein of TNBS-induced colitis rat model was significantly decreased compared with the normal control group (P < 0.05). Treatment with SQR or mesalazine significantly increased the expression of c-kit protein compared with the administration of control formulations (P < 0.05), especially the high-dose SQR group.</p><p><b>CONCLUSION</b>SQR could alleviate and repair the injured ICC, and improve its quantity, which might be involved in regulating intestinal motility.</p>


Subject(s)
Animals , Colitis , Drug Therapy , Pathology , Colon , Cell Biology , Metabolism , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Interstitial Cells of Cajal , Pathology , Male , Medicine, Chinese Traditional , Mesalamine , Therapeutic Uses , Peroxidase , Metabolism , Proto-Oncogene Proteins c-kit , Metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Trinitrobenzenesulfonic Acid
13.
Article in Chinese | WPRIM | ID: wpr-235136

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the feasibility of peptide mass fingerprinting for non-invasive differential diagnosis of IgA nephropathy (IgAN) from the non-IgA nephropathy (IgAN).?</p><p><b>METHODS</b>According to the results of renal biopsy, 56 patients were divided into IgAN group (n=28) and non-IgAN group (n=28), and peptide mass fingerprints were acquired from these patients using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).</p><p><b>RESULTS</b>Nine different peptides were identified between IgAN and non-IgAN. The two most distinctive differentially expressed peptides, with peptide peak values of 4476.46 and 1968.10, showed area under curve values of 86.18% and 79.77%. Principal component analysis demonstrated that the accumulated explained variance of the first 8 differential peptides reached 95%, suggesting the feasibility of differential diagnosis of IgAN from non-IgAN. Comparison with the Matrix protein database identified the peptide with a relative molecular mass of 5338.08 as a fragment of mucin 4 inform and the 2082.77 peptide as fragment of α1-II type collagen inform.</p><p><b>CONCLUSION</b>MALDI-TOF MS is feasible for differential diagnosis of IgAN and non-IgAN and also has great potentials in the classification of the subtypes of other systemic diseases.</p>


Subject(s)
Adult , Diagnosis, Differential , Feasibility Studies , Female , Glomerulonephritis, IGA , Diagnosis , Humans , Kidney Diseases , Diagnosis , Male , Middle Aged , Peptide Mapping , Methods , Peptides , Chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Methods , Young Adult
14.
Article in Chinese | WPRIM | ID: wpr-336184

ABSTRACT

<p><b>OBJECTIVE</b>To construct a vector expressing small interfering RNA (siRNA) against Rac1 gene and observe its effect on soft agar colony formation of SW480 cells in vitro.</p><p><b>METHODS</b>Oligos of 64 base pairs for hairpin RNA targeting Rac1 were chemically synthesized and annealed. The siRNA constructs for Rac1, produced by inserting the annealed oligos into the downstream of H1 promoter of linearized pSUPER, were confirmed by restriction digestion and DNA sequencing. The constructed Rac1-siRNA was transfected into SW480 cells and Western blotting was performed to assess the expression and interference efficiency of siRNAs against Rac1.The soft agar colony formation assay was used to study the effect of Rac1 gene silencing on SW480 cells.</p><p><b>RESULTS</b>Restriction digestion and DNA sequencing showed that the siRNA targeting Rac1 gene was successfully constructed. The siRNA could effectively down-regulate the expression of Rac1 in SW480 cells. Soft agar colony formation assay showed that the colony number and diameter of SW480 cells was reduced after siRNA transfection.</p><p><b>CONCLUSION</b>A vector expressing hairpin RNA against Rac1 gene are successfully produced, which significantly reduces the colony numbers and size of SW480 cells in vitro, suggesting that Rac1 plays an important role in the growth of colorectal cancer in vitro.</p>


Subject(s)
Base Sequence , Cell Line, Tumor , Cell Proliferation , Colonic Neoplasms , Pathology , Down-Regulation , Humans , Molecular Sequence Data , RNA Interference , RNA, Small Interfering , Genetics , Transfection , rac1 GTP-Binding Protein , Genetics
15.
Article in Chinese | WPRIM | ID: wpr-289960

ABSTRACT

<p><b>OBJECTIVE</b>To study the metabolic difference of body influenced by active stress and passive stress under special events.</p><p><b>METHODS</b>To detect serum multiple biochemistry index of 57 earthquake rescue medical team and 13 victims of a natural calamity in Wenchuan earthquake by using Hitachi 7600 automatic analyzer.</p><p><b>RESULTS</b>Stress affected biochemistry index deeply. To compared with rescue medical team, the serum ADA, ALP and TG of victims increased obviously and TP, ALB, MAO, Cr, UA, K, Na, Cl, Ca, ApoA1 and HDL decreased obviously.</p><p><b>CONCLUSION</b>Many biochemistry index have been changed under stress and it relate with stress extent. The human body function status was better in active stress than in passive stress.</p>


Subject(s)
Blood Chemical Analysis , China , Disasters , Earthquakes , Humans , Metabolism , Physiology , Rescue Work , Stress, Physiological , Physiology
16.
Article in Chinese | WPRIM | ID: wpr-268765

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of p21-activated kinase-1 (PAK1) gene transfection on the invasiveness of human colorectal carcinoma SW480 cells in vitro.</p><p><b>METHODS</b>SW480 cells in routine culture were transfected with the recombinant plasmid EGFP-C1/PAK1 via Lipofectamine(TM) 2000. The expression of PAK1 protein in SW480 cells was detected using Western blotting, and the changes of the invasiveness of SW480 cells were evaluated using Boyden chamber invasion assay.</p><p><b>RESULTS</b>Forty-eight hours after transfection with pEGFP-C1/ PAK1, the PAK1 protein expression increased significantly in comparison with those in negative and vector control groups. The invasiveness of the SW480 cells was significantly enhanced after the transfection.</p><p><b>CONCLUSION</b>The PAK1 gene transfection can increase the expression of PAK1 in SW480 cells and enhance the invasiveness of the cells. PAK1 can be associated with the invasiveness and metastasis of colorectal carcinoma cells.</p>


Subject(s)
Cell Line, Tumor , Colorectal Neoplasms , Genetics , Pathology , Gene Expression , Genetic Vectors , Humans , Neoplasm Invasiveness , Neoplasm Metastasis , Plasmids , Transfection , p21-Activated Kinases , Genetics
17.
Article in Chinese | WPRIM | ID: wpr-233702

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of X-ray exposure on the release of soluble tumor necrosis factor receptor-p75 (sTNFR-p75) in hepatocellular carcinoma HepG2 cells in vitro.</p><p><b>METHODS</b>Enzyme-linked immunosorbent assay (ELISA) was used to examine the levels of sTNFR-p75 in the supernatants of HepG2 cells before and after X-ray exposure. The cell apoptosis was analyzed by flow cytometry and transmission electron microscope(TEM), and the morphological changes of the cells were examined under optical microscope and transmission electron microscope(TEM).</p><p><b>RESULTS</b>X-ray exposure of the cells resulted in a strong increase of cell apoptosis (P<0.05) and sTNFR-p75 production in the cells as compared with the those before the exposure (P<0.01). Optical microscopy revealed apoptotic changes of HepG2 cell after the exposure, shown as cell shrinkage, spherical cell morphology, cytoplasmic and nuclear condensation. Apoptotic bodies were detected by TEM.</p><p><b>CONCLUSION</b>X-ray exposure induces HepG2 cells apoptosis by inhibiting the release of sTNFR-p75 into the supernatant.</p>


Subject(s)
Animals , Apoptosis , Radiation Effects , Carcinoma, Hepatocellular , Pathology , Bodily Secretions , Cell Line, Tumor , Culture Media, Conditioned , Chemistry , Metabolism , Radiation Effects , Humans , Liver Neoplasms , Pathology , Bodily Secretions , Microscopy , Receptors, Tumor Necrosis Factor, Type II , Chemistry , Bodily Secretions , Solubility , X-Rays
18.
Article in Chinese | WPRIM | ID: wpr-280069

ABSTRACT

<p><b>OBJECTIVE</b>To clone migfilin-N terminal sequence into E.coli and obtain a fusion protein for preparing rabbit polyclonal antibody against migfilin, thereby facilitating the study of the role of migfilin in the biological behavior of colon cancer.</p><p><b>METHODS</b>Based on human migfilin cDNA sequence, a pair of primers was designed to amplify migfilin-N terminal sequence by PCR. The PCR product was subcloned into the bacterial expression vector pGEX-4T-1 with EcoRI/XhoI sites, and the target recombinant plasmids were identified with enzymatic cleavage followed by DNA sequence analysis. By transforming the expression vector into component E.coli BL(21) cells, the GST-migfilin-N fusion protein was expressed with IPTG induction. Glutathione-sepharose beads were used to purify the fusion protein, and anti-migfilin polyclonal antibody was produced by immunization of rabbits with the purified GST-migfilin N-terminal fusion protein. The resultant anti-migfilin polyclonal antibody was purified by protein A beads and used for Western blotting for detecting migfilin expression in different cell lines.</p><p><b>RESULTS AND CONCLUSION</b>The migfilin-N terminal gene fragment was cloned successfully, and purified GST-migfilin N-terminal fusion protein and anti-rabbit migfilin polyclonal antibodies were obtained. Western blot analysis demonstrates that the antibodies specifically detected migfilin expression in the cell lines, which may facilitate further investigation of the role of migfilin in the biology of colon cancer.</p>


Subject(s)
Animals , Antibodies, Monoclonal , Allergy and Immunology , Base Sequence , Blotting, Western , Cell Adhesion Molecules , Genetics , Allergy and Immunology , Cell Line, Tumor , Cloning, Molecular , Colonic Neoplasms , Genetics , Metabolism , Pathology , Cytoskeletal Proteins , Genetics , Allergy and Immunology , DNA, Complementary , Chemistry , Genetics , Escherichia coli , Genetics , Humans , Molecular Sequence Data , Rabbits , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
19.
Microbiology ; (12)2008.
Article in Chinese | WPRIM | ID: wpr-685981

ABSTRACT

Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.

20.
Article in Chinese | WPRIM | ID: wpr-337352

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of short hairpin RNA (shRNA) targeting survivin on adhesion and invasion of human colon carcinoma cell line SW480 in vitro.</p><p><b>METHODS</b>According to the sequence of the coding region of survivin gene, two strings of 19 nucleotides of inverted sequence flanking the loop sequence of two complementary 9-base oligonucleotides were designed and synthesized to prepare the hairpin construct as the DNA templates for the target shRNA. The shRNA templates were cloned into shRNA expression vector pRNAT-U6.1/Neo, and the resulted vector pRNAT-U6.1/Neo-survivin was transfected into SW480 cells using Lipofectamine 2000. Western blotting was performed to evaluate survivin gene silencing induced by shRNA transfection at the protein level, and the biological behaviors of the SW480 cells were investigated by cell-matrix adhesion, invasion and gelatin-zymography assays.</p><p><b>RESULTS</b>Western blotting revealed significantly lowered survivin protein expression in transfected SW480 cells, and survivin gene silencing induced by shRNA significantly suppressed the metastatic potential of SW480 cells in association with suppressed MMPs activity.</p><p><b>CONCLUSIONS</b>Survivin may play an important role in modulating human colorectal carcinoma cell invasion and metastasis, and survivin gene silencing can inhibit human colorectal cancer cell invasion and the production of MMP-2 and MMP-9. Survivin may affect invasion and metastasis of human colorectal carcinoma cells via regulating the production of MMPs.</p>


Subject(s)
Animals , Blotting, Western , Cell Line, Tumor , Colonic Neoplasms , Genetics , Pathology , Gene Silencing , Humans , Inhibitor of Apoptosis Proteins , Inverted Repeat Sequences , Matrix Metalloproteinases , Bodily Secretions , Microtubule-Associated Proteins , Genetics , Neoplasm Invasiveness , Genetics , Neoplasm Metastasis , Genetics , RNA, Small Interfering , Genetics
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