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1.
Chinese Journal of Orthopaedics ; (12): 831-840, 2023.
Article in Chinese | WPRIM | ID: wpr-993510

ABSTRACT

Objective:Observing the effect of exosomes derived from hypoxic Bone marrow mesenchymal stem cells (BMSCs) on the function of chondrocytes, and exploring the role and mechanism of exosomal miR-196b-5p. Evaluating the application prospects of hypoxic BMSCs exosomes and miR-196b-5p for cartilage regeneration.Methods:Chondrocytes were cultured in the supernatant of BMSCs cultured under normoxia or hypoxia, respectively. The proliferation of chondrocytes was detected by CCK-8 assay and the expressions of Collagen type 2 (Col2), Col1, Aggrecan and SOX9 were detected by qPCR to evaluate the effect of hypoxic BMSCs paracrine on chondrocyte functions. Obtaining normoxic and hypoxic exosomes through ultracentrifugation, and testing their effects on the proliferation and anabolic-related genes of chondrocytes through CCK-8 assay and qPCR. Verifying the expression of miR-196b-5p in hypoxic exosomes based on exosomal miRNA array. Knocking out miR-196b-5p in hypoxic BMSCs, and detecting the effect of hypoxic exosomal miR-196b-5p on the functions of chondrocytes by loss-of-function assay. Predicting the downstream of miR-196b-5p through bioinformatics tools, and exploring the mechanism of hypoxic exosomal miR-196b-5p by gain-of-function assays. Hypoxic exosomes and miR-196b-5p-knockout hypoxic exosomes were loaded on silk fibroin hydrogel and subcutaneously into nude mice. After 4 weeks of culture, histological staining of saffron O, Masson and biochemical content of sGAG and collagen were performed to assess the application prospect of hypoxic exosomes and hypoxic exosomal miR-196b-5p on cartilage regeneration. Results:The results of CCK-8 assay and qPCR indicated that the supernatant of hypoxic BMSCs significantly promoted the proliferation of chondrocytes 1.20±0.07 and the expression of cartilage-related markers (Col2 2.95±0.17, Aggrecan 2.45±0.27, SOX9 2.92±0.29) compared to normoxic BMSCs (0.94±0.04, 1.89±0.09, 1.67±0.21, 1.76±0.16), the differences were statistically significant ( P<0.05). The result of CCK-8 assay showed that hypoxic exosomes (1.28±0.04) promoted the proliferation of chondrocytes compared to normoxic exosomes 1.05±0.06, the differences were statistically significant ( P<0.05). CCK-8 assay revealed that the down-regulation of miR-196b-5p in hypoxic exosomes 0.99±0.06 attenuated the proliferation of chondrocytes compared to control group 1.20±0.07, the differences were statistically significant ( P<0.05); the expression of Col2 0.56±0.04, Aggrecan 0.74±0.09, and SOX9 0.45±0.05 in chondrocytes was reduced in the miR-196b-5p knockdown group compared to the control group (1.00±0.09, 1.00±0.12, 1.00±0.07), the differences were statistically significant ( P<0.05). Co-transfection of pmirGLO-BACH1-WT reporter vector with miR-196b-5p mimics decreased the luciferase activity 0.73±0.06, the differences were statistically significant ( P<0.05). Co-transfection of pmirGLO-BACH1-MUT reporter vector with miR-196b-5p mimics showed no change in luciferase activity. BACH1 is the target of miR-196b-5p. Subcutaneous culture in nude mice showed that hypoxic exosomes significantly promoted the deposition of sGAG 383.2±21.54 and collagen 67.40±3.45, while reducing the expression of miR-196b-5p in hypoxic exosomes weakened the deposition of sGAG 258.4±19.50 and collagen 57.15±4.95, the differences were statistically significant ( P<0.05). Conclusion:Hypoxic exosomes promoted the functions of chondrocytes by inhibiting the expression of BACH1 through miR-196b-5p. Hypoxic exosomes can be applied in cartilage regeneration.

2.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery ; (12): 550-555, 2023.
Article in Chinese | WPRIM | ID: wpr-982784

ABSTRACT

Objective:To observe the efficacy and safety of the M receptor antagonist Bencycloquidium bromide nasal spray in treatment of seasonal allergic rhinitis with runny nose as the main symptom. Methods:From August 2021 to September 2021, 134 patients with seasonal allergic rhinitis were enrolled in the otolaryngology Outpatient Department of Peking University Third Hospital, First Affiliated Hospital of Harbin Medical University and China-Japanese Friendship Hospital of Jilin University, including 71 males and 63 females, with a median age of 38 years. TNSS score and visual analogue scale(VAS) of total nasal symptoms were observed during 2 weeks of treatment with Bencycloquidium bromide nasal spray. Results:TNSS score decreased from (8.89±3.31) on day 0 to (3.71±2.51) on day 14(P<0.001), VAS score of nasal symptoms decreased from (24.86±7.40) on day 0 to (6.84±5.94) on day 14(P<0.001), VAS score of rhinorrhoea decreased from (6.88±2.06) on day 0 to (1.91±1.81) on day 14(P<0.001). Rhinoconjunctivitis quality of life questionnaire(RQLQ) score decreased from (94.63±33.35) on day 0 to (44.95±32.28) on day 14(P<0.001). The incidence of adverse reaction was low and no serious adverse events occurred during the whole experiment. Conclusion:Bencycloquidium bromide nasal spray has significant efficacy and good safety in the treatment of seasonal allergic rhinitis.


Subject(s)
Male , Female , Humans , Adult , Rhinitis, Allergic, Seasonal/drug therapy , Nasal Sprays , Quality of Life , Administration, Intranasal , Rhinorrhea , Double-Blind Method , Treatment Outcome , Rhinitis, Allergic/drug therapy
3.
Chinese Journal of Orthopaedics ; (12): 1073-1080, 2021.
Article in Chinese | WPRIM | ID: wpr-910693

ABSTRACT

Objective:A new type of bio-ink and polycaprolactone (PCL) were used to construct an integrated osteochon-dral composite tissue block by multi-nozzle 3D bioprinter. And the repair results to osteochondral defects were evaluated.Methods:In freeze-drying group: Freeze-dried composite scaffold made by silk fibroin (SF) and β-tricalcium phosphate was used to repair osteochondral defects, as control. In the 3D printing group: PCL was used to printed a hollow multi-layer cylinder frame by 3D biological printer. Extracellular matrix, SF and bone marrow mesenchymal stem cells were used as chondral bio-ink. Then, chon-dral bio-ink was used to print tissue-engineered cartilage on top of PCL frame. Before implantation of cartilage defect, autogenous cancellous bone was filled in PCL frame, then the tissue-engineered osteochondral composite was used to repair osteochondral defects. In mosaic group: Autologous osteochondral transplantation was performed. The repair results of the above three groups were compared by histological score, biochemical analysis and biomechanical test to evaluate the effect of repairing rabbit cartilage defects.Results:The compression modulus of neo-cartilage in the 3D print group 2.56±0.30 MPa was close to that of the mosaic group 2.51±0.13 MPa ( P>0.05), and significantly higher than that of freeze-dried group 1.37±0.14 MPa ( F=11.058, P<0.05). The sGAG contents in the 3D print group 14.49±0.7 μg/mg was close to that of the mosaic group 14.98±0.81 μg/mg ( P>0.05), and significantly higher than that of freeze-dried group 8.72±0.73 μg/mg ( F=20.973, P<0.05). However, there was no significant difference in collagen content between the three groups ( P>0.05). The results of ICRS cartilage repair histology score showed that the scores of the 3D print group were close to those of the mosaic group in the matrix, cell distribution, cell viability and subchondral bone ( P>0.05), and were significantly higher than those of freeze-dried group in the surface and cartilage mineralization scores ( F=19.544, P<0.05). Conclusion:Using the new bio-ink to make bone cartilage composite scaffold by 3D bio printing can simplify the construction of tissue-engineered bone cartilage composite tissue in vitro, and can repair cartilage defects in vivo.

4.
Chinese Journal of Practical Surgery ; (12): 719-721, 2019.
Article in Chinese | WPRIM | ID: wpr-816454

ABSTRACT

OBJECTIVE: To discuss whether grade B pancreatic fistula after pancreaticoduodenectomy(PD)can be graded according to the severity and to provide reference for accurate evaluation and treatment.METHODS: The clinical data of 343 patients underwent PD surgery in the First Affiliated Hospital of Harbin Medical University from December2012 to June 2016 were retrospectively collected,among which 72 cases followed grade B pancreatic fistula. Grade B pancreatic fistula after operation was divided into severe groups of which imaging-assisted interventional therapy was needed and mild group. The total medical cost,length of hospital stay,drainage time,ICU transfer,introperitoneal effusion,introabdominal infection,abdominal hemorrhage,biliary fistula,delayed gastric emptying,number of types and incidence of complications except pancreatic fistula,incidence of pancreatic fistula as the most serious complications,postoperative morbidity index(PMI),fistula average complication burden(ACB)were statistically analyzed between two groups.RESULTS: Univariate analysis revealed that there were significant differences between two groups in the total medical cost(84,000 yuan vs. 132,000 yuan),length of hospital stay(29.0 days vs. 42.0 days),drainage time(20.5 d vs.53.0 d),introperitonal effusion rate(41.1% vs. 87.5%),introabdominal infection rate(10.7% vs. 43.8%),abdominal hemorrhage rate(7.1% vs. 56.3%),the number of other types of complications except pancreatic fistula(1 vs. 4),pancreatic fistula as the most serious complications(53.6% vs. 87.5%),PMI(0.22±0.08 vs. 0.37±0.00),ACB(0.19±0.08 vs. 0.37±0.00),and the P-values were less than 0.05 respectively.CONCLUSION: The severity of grade B pancreatic fistula of ISGPS update classification is heterogeneous,which can be divided into mild and severe groups. It provides a reference for precise and individualized treatment for grade B pancreatic fistula.

5.
Chinese Journal of Tissue Engineering Research ; (53): 1762-1767, 2018.
Article in Chinese | WPRIM | ID: wpr-698610

ABSTRACT

BACKGROUND: Hallux valgus is a common orthopedic disease, and its causes are complex and treatment is varied. The mechanical analysis of hallux valgus is an issue of concern. The finite element analysis makes it predictable to treat hallux valgus. OBJECTIVE: To explore the clinical application of finite element analysis in biomechanical study of hallux valgus.METHODS: The first author searched CNKI and PubMed databases from January 1980 to March 2017 using the key words of "finite element, hallux valgus" in English and Chinese, respectively. The repetitive, irrelevant and low-quality articles were excluded. Finally 33 eligible articles were included in accordance with the inclusion criteria, and the critical issues of finite element analysis applied in hallux valgus were reviewed. RESULTS AND CONCLUSION: (1) There are many researches concerning finite element of hallux valgus, which mostly require physicians to work with engineers. These methods are already very mature, but most of the model and material properties of the data come from foreign researches. (2) The finite element analysis is important and reliable for the etiology of hallux valgus, preoperative planning and prognosis. (3) The finite element model of the hallux valgus is only used on static analysis and gait cycle analysis, the modeling details and definition of material properties still need to be improved.

6.
Journal of Peking University(Health Sciences) ; (6): 603-606, 2016.
Article in Chinese | WPRIM | ID: wpr-496206

ABSTRACT

Objective:To assess the discrepancy between preoperative needle biopsy (NB)Gleason score and pathological specimen Gleason score (GS)after radical prostatectomy,and to explore the risk factors of postoperative upgrading of GS.Methods:We retrospectively evaluated 160 patients who suf-fered from biopsy proved prostatic carcinoma and performed radical prostatectomy.Age of the patients was 57 -82 years,with the average age of 71.6;prebiopsy prostate specific antigen (PSA)was 0.31 -40.32 μg/L,with the average PSA of 11.29 μg/L;body mass index (BMI)was 16.41 -32.04 kg/m2 , with the average BMI of 23.63 kg/m2;prostate volume (PV)was 9.52 -148.46 mL,with the average PV of 40.19 mL.All the patients included in the study had complete information for clinical variables, including age,BMI,prebiopsy PSA level,PV,number of biopsy cores obtained,percentage,clinical stage,and biopsy GS.Grading of NB Gleason score was compared with their corresponding radical pros-tatectomy specimens,and the discrepancy between the NB and prostatectomy specimens GS assessed. Upgrading was defined as any increase in the pathological GS over that of the biopsy GS as a total sum of primary and secondary grades or a change in the order of primary and secondary grades towards higher ones.Univariable and multivariable Logistic regression analyses were used to identify predictors of patho-logical grading changes.Results:Of the 160 patients,the specimen GS was upgraded in 49 (30.6%) patients and remained with no change in 82 (51.3%)patients.Univariate and multivariate regression analysis showed that prostate volume and biopsy GS were independent predictors with postoperative upgra-ding of GS.Age,BMI,PSA before needle biopsy,clinical stage and needle number showed no statistical significance (P >0.05).Conclusion:Lower biopsy GS and smaller prostate volume are increased risks for clinically upgrading of GS after radical prostatectomy.This fact should be kept in mind when deciding on therapy decisions for patients with prostate cancer.

7.
Journal of Central South University(Medical Sciences) ; (12): 471-476, 2016.
Article in Chinese | WPRIM | ID: wpr-815012

ABSTRACT

OBJECTIVE@#To investigate the correlation between cyclin-dependent kinase inhibitor p27kip1 and trastuzumab-resistance in gastric cancer.
@*METHODS@#We selected HER2-overexpressed human gastric cancer cell line NCI-N87 to establish trastuzumab-resistant NCI-N87/TR cell line by stepwise exposure to different doses of trastuzumab. The 50% inhibitory concentration (IC(50)) of trastuzumab and resistance index (RI) were calculated or analyzed by MTT assay. The expression levels of cdk2 and p27kip1 were detected by Western blot. After the treatment with cdk2 inhibitor (Purvalanol A), the expression levels of relevant proteins in NCI-N87/TR cells were detected by Western blot, and the sensitivity to trastuzumab was analyzed by MTT assay. 
@*RESULTS@#Compared with NCI-N87 cells, the expression of cdk2 was significantly increased in NCI-N87/TR cells (P<0.001), while the expression of p27kip1 showed a significant decrease (P<0.001). Restoration of the p27kip1 protein expression by cdk2 inhibitor (Purvalanol A) increased the sensitivity of NCI-N87/TR to trastuzumab.
@*CONCLUSION@#Down-regulation of p27kip1 might be a mechanism for triggering trastuzumab resistance to gastric cancer cell line NCI-N87.


Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Cell Line, Tumor , Cyclin-Dependent Kinase 2 , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p27 , Genetics , Metabolism , Drug Resistance, Neoplasm , Purines , Pharmacology , Stomach Neoplasms , Genetics , Metabolism , Pathology , Trastuzumab , Pharmacology
8.
Journal of Guangzhou University of Traditional Chinese Medicine ; (6): 720-724,728, 2015.
Article in Chinese | WPRIM | ID: wpr-603283

ABSTRACT

Objective To observe the effect of Professor Deng Tietao’s Brain-strengthening Recipe 1 on learning and memory abilities of rats with vascular dementia, and to explore its molecular biology mechanism. Methods The vascular dementia rat model was established by permanent occlusion of the bilateral arteria carotis communis. Sixty male SD rats were randomly divided into 5 groups, namely sham-operation group, model group, nimodipine (5.4 mg·kg-1·d-1) group, and high-and low-dose Chinese medicine groups (57.6 and 14.4 g·kg-1·d-1, respecitvely) . After medication for 30 days, the learning and memory abilities were evaluated by Morris water maze test. The histological changes of the rat hippocampal tissues were observed after HE staining, and the mRNA expression of N-methyl-D-aspartate receptor subunit 2B ( NR2B) in rat hippocampus was measured by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) . Results The water maze test results showed that the escape latency of the rats in the sham-operation group was significantly decreased on the third day, and was significantly decreased in the high-dose Chinese medicine group on the fourth day ( P0.05). HE staining results for the model group were presented as the absence of neural cells in hippocampus CA1 region, karyorrhexis of nerve cells, indistinctness of nuclear membrane, disappearance of nucleolus, shrinkage of nerve cells, increase of cytoplasm acidophily, and reactive hyperplasia of astrocytes. And in the hippocampal CA1 region of high- and low-dose Chinese medicine groups, the injured neural cells were found, and the lesions were slighter than those of the model group. Compared with the model group, the mRNA expression of NR2B was increased in nimodipine group and high-dose Chinese medicine group ( P0.05). Conclusion Brain-strengthening Recipe 1 has obvious effects on improving the learning and memory abilities of vascular dementia rats, and the therapeutic mechanism is associated with reduction of the degree of neuronal damage in hippocampus and with the upregulation of the mRNA expression of NR2B.

9.
International Journal of Surgery ; (12): 240-243,289, 2015.
Article in Chinese | WPRIM | ID: wpr-601641

ABSTRACT

Objective To study and evaluate mini-titanum-plates the effects applied to four limbs bone and joint fracture.Methods 132 patients in our hospital suffered from four limbs fractures treated with open reduction internal fixation of Medicon mini-plate from February 2004 to June 2013 were studied retrospectively,including 60 cases of metacarpal and phalanx fracture(93 lesions),37 cases of metatarsal and phalanx fracture(62 lesions),15 cases of radial head fracture,9 cases of scaphoid fracture,4 cases of coronoid process of ulna fracture,4 cases of patella frature,3 cases of condyles of femur.Results Clinical outcomes:hands and feet group:follow-up postoperative 8-12 months,evaluation results showed that excellent was 78cases,good was 66cases,poor was 11cases,good rate was 92.9%.group of radius head fracture:Fractures got osteal union after 6 months to 1 year.good rate was 93.3%.group of scaphoid bone fracture:Fractures got osteal healing.all cases adopt improved Mayo Evaluation Wrist Function to evaluate,good rate was 88.9%.Including 4 cases of Coronoid process of ulna fracture,4 cases of patella fracture,3 cases of femoral condylar fracture,the excellent good rates was 100%.Conclusions Metacarpal and metatarsals phalanx fracture treated with mini-plate can get reduction anatomically,control rotation,prevent malformation and keep compression,they are the ideal internal fixation material;free fragments for intra-articular fracture fixed by mini-titanum-plate and screws can get reduction precisely,be fixed satisfactory,also can choose the appropriate application.

10.
China Journal of Orthopaedics and Traumatology ; (12): 453-457, 2014.
Article in Chinese | WPRIM | ID: wpr-301794

ABSTRACT

<p><b>OBJECTIVE</b>To explore effective approaches of treating elderly patients with distal tibiofibular fractures.</p><p><b>METHODS</b>From August 2008 to October 2012,175 elderly patients with distal tibiofibular fractures were treated with locking compression plate (LCP) through anterior tibial. There were 112 males and 63 females with an average of 71.3 (ranged 60 to 83) years old. Of them,89 cases were treated by anterior tibial tension reduced incision with LCP,including 62 males and 27 females with a mean age of (71.8 +/- 6.4) years old. Eighty-six patients were treated by distal tibial incision with LCP,including 58 males and 28 females with a mean age of (70.3 +/- 6.7) years old. Swelling time, operation time, intraoperative blood loss, hospital stay, healing time, complications and AOFAS scores were compared between two groups after operation.</p><p><b>RESULTS</b>Swelling time in anterior tension reduced incision with LCP and distal tibial incision with LCP was (5.6 +/- 1.3) and (9.7 +/- 2.1) days, healing time was (4.2 +/- 1.4) and (5.4 +/- 1.9) months,and complications were found 3 in tension reduced incision and 10 in distak tibial incision respectively;and all data shown statistically significant differences between two groups (P < 0.05). At 12 months after operation,AOFAS score was 89.0 +/- 9.7, 87.9 +/- 9.4; and there was no statistically significant difference between two groups (P > 0.05).</p><p><b>CONCLUSION</b>Tension reduced incision through anterior tibial combined with locking compression plate fixation in treating elderly patients with distal tibiofibular fractures can provide good clinical effects with quick fracture healing and low complications.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bone Plates , Fibula , Wounds and Injuries , General Surgery , Fracture Fixation, Internal , Methods , Minimally Invasive Surgical Procedures , Tibia , Wounds and Injuries , General Surgery , Tibial Fractures , General Surgery , Treatment Outcome
11.
Journal of Southern Medical University ; (12): 474-478, 2013.
Article in English | WPRIM | ID: wpr-322019

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expressions of different forms of ROS fusions in Chinese patients with cholangiocarcinoma (CCA).</p><p><b>METHODS</b>RT-PCR was employed to examine formalin-fixed and paraffin-embedded CCA samples from stage I-IV patients for detection of ROS fusions involving Fused in Glioblastoma (FIG), solute carrier protein (SLC34A2) and major histocompatibility complex class II invariant chain (CD74). Serpin peptidase inhibitor clade A member 1 (SERPINA1) was detected as the reference gene.</p><p><b>RESULTS</b>In all the 56 CCA samples, 80.4% (45/56) were positive for SERPINA1 expression as evaluable samples. Of these evaluable samples, none expressed the ROS fusions.</p><p><b>CONCLUSION</b>ROS fusions are not common in Chinese CCA patients.</p>


Subject(s)
Female , Humans , Male , Middle Aged , Antigens, Differentiation, B-Lymphocyte , Genetics , Metabolism , Bile Duct Neoplasms , Metabolism , Pathology , Carrier Proteins , Genetics , Metabolism , Cholangiocarcinoma , Metabolism , Pathology , Gene Expression , Histocompatibility Antigens Class II , Genetics , Metabolism , Membrane Proteins , Genetics , Metabolism , Oncogene Proteins, Fusion , Genetics , Metabolism , Paraffin Embedding , Protein-Tyrosine Kinases , Genetics , Metabolism , Proto-Oncogene Proteins , Genetics , Metabolism , Sodium-Phosphate Cotransporter Proteins, Type IIb , Genetics , Metabolism
12.
Journal of Southern Medical University ; (12): 312-316, 2012.
Article in Chinese | WPRIM | ID: wpr-267610

ABSTRACT

<p><b>OBJECTIVE</b>To establish a gastric cancer cell line with stable expression of metastasis-associated in colon cancer 1 (MACC1) and detect the changes in tumor-related gene expression profiles for investigating the possible regulation mechanisms between MACC1 and the differentially expressed genes.</p><p><b>METHODS</b>The full-length MACC1 cDNA was amplified from human embryonic kidney 293FT cells and cloned into the pBaBb-puro vector. The recombinant pBaBb-puro-MACC1 expression vector, after identification with restriction enzyme digestion, was transfected into 293FT cells, and the expression of fluorescent reporter gene was observed. pBaBb-puro-MACC1 vector was transfected into human gastric cancer BGC-823 cell line to establish BGC-823/pBaBb-puro-MACC1 cell line stably expressing MACC1. Quantitative RT-PCR and Western blotting were used to detect MACC1 expression in both BGC-823/pBaBb-puro-MACC1 and control BGC-823 cells. High-throughout cDNA microarray was used to screen the effects of MACC1 on the gene expression profiles of gastric cancer cells.</p><p><b>RESULTS</b>The recombinant pBaBb-puro-MACC1 plasmid was successfully constructed and verified by PCR and sequencing. BGC-823/pBaBb-puro-MACC1 cells showed significantly increased MACC1 mRNA expression as compared with the control cells. The results of cDNA microarray identified 33 up-regulated and 24 down-regulated genes in the cells after MACC1 transfection involved were in various cellular functions.</p><p><b>CONCLUSION</b>The established BGC-823/pBaBb-puro-MACC1 gastric cancer cell line show some important molecular changes caused by MACC1.</p>


Subject(s)
Humans , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Genetics , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , HEK293 Cells , Oligonucleotide Array Sequence Analysis , Stomach Neoplasms , Metabolism , Pathology , Transcription Factors , Genetics , Metabolism , Transcriptome , Transfection
13.
China Journal of Orthopaedics and Traumatology ; (12): 654-657, 2012.
Article in Chinese | WPRIM | ID: wpr-321891

ABSTRACT

<p><b>OBJECTIVE</b>To measure the stability of Evans procedure and Chrisman-Snook technique in the treatment of II degree lateral collateral ligament of ankle joint, and provide basis for treatment and prognosis.</p><p><b>METHODS</b>From July 2008 to June 2009,18 frozen corpes were collected, including 10 males and 8 females, with an average age of fresh 39.3 +/- 11.2 years. The frozen corpes were randomly divided into three group, including normal controls(group A), Evans procedure (group B) and Chrisman-Snook technique ( group C), 6 specimens in each group. Anterior talofibular ligament and calcaneofibular ligament were cut off to cause II degree lateral collateral ligament in group B and C. Evans procedure or Chrisman-Snook technique were applied to restore lateral collateral ligament, and measure biomechnics. The displacement of tibiotalar joint and subtalar joint were observed.</p><p><b>RESULTS</b>(1) The lateral stress results of tibiotalar joint showed the displacement by Evans procedure (group B) was greater than other groups (P < 0.0001). There were no significant differences between group A and C (P > 0.05). (2) The lateral stress results of subtalar joint showed the displacement by Evans procedure (group B) was greater than other groups (P< 0.0001). There were no significant differences between group A and C (P > 0.05).</p><p><b>CONCLUSION</b>Ankle instability is caused by ankle joint lateral collateral ligament injury. Chrisman-Snook technique is better than Evans procedure in stability on the early stage of ankle joint restoration, and conform to principle of biomechanics.</p>


Subject(s)
Adult , Female , Humans , Male , Ankle Joint , Biomechanical Phenomena , Lateral Ligament, Ankle , Diagnostic Imaging , Wounds and Injuries , General Surgery , Mechanical Phenomena , Prognosis , Radiography , Plastic Surgery Procedures , Methods
14.
Chinese Journal of Oncology ; (12): 574-578, 2011.
Article in Chinese | WPRIM | ID: wpr-320167

ABSTRACT

<p><b>OBJECTIVE</b>To explore the changes of the sensitivity of cetuximab-resistant human nasopharyngeal carcinoma (hNPC) cells 5-8F/Erbitux to cetuximab by silencing H-ras gene with RNA interference (RNAi).</p><p><b>METHODS</b>The 5-8F/Erbitux cells were induced by stepwise exposure to increasing doses of cetuximab. Western blot was conducted to detect the protein levels of H-ras and K-ras. Real-time PCR was employed to detect the expression of H-ras and K-ras. H-ras-shRNA plasmids (shRNA vector carrying the H-ras gene) were constructed and transferred into 5-8F/Erbitux cells. The gene and protein expression levels of H-ras and the changes of the sensitivity of 5-8F/Erbitux cells to cetuximab after transfection were measured, respectively.</p><p><b>RESULTS</b>After treatment with cetuximab for 3 and 5 days, the resistance index (RI) of the 5-8F/Erbitux cells was 1.2 and 1.1, and the protein levels of H-ras and K-ras in 5-8F/Erbitux cells were 0.798 +/- 0.019 and 0.190 +/- 0.011, respectively, significantly higher than that in the 5-8F cells (P<0.001). The gene expressions of H-ras and K-ras in 5-8F/Erbitux cells were 1.260 +/- 0.114 and 0.850 +/- 0.006, respectively. Compared with 5-8F cells, the former was higher (P = 0.016) and the latter was lower (P = 0.000). After transfection with H-ras-shRNA plasmid, the 5-8F/Erbitux cells showed reduced levels of H-ras gene and protein, and the cell apoptosis and inhibition rates increased significantly (P<0.001).</p><p><b>CONCLUSIONS</b>H-ras siRNA can reverse cetuximab-resistance of 5-8F/Erbitux cells through down-regulation of H-ras gene expression, indicating that the generation of cetuximab-resistance in 5-8F/Erbitux cells is associated with amplification and overexpression of the H-ras gene.</p>


Subject(s)
Humans , Antibodies, Monoclonal , Pharmacology , Antibodies, Monoclonal, Humanized , Antineoplastic Agents , Pharmacology , Apoptosis , Carcinoma , Cell Line, Tumor , Cell Proliferation , Cetuximab , Drug Resistance, Neoplasm , Gene Expression Regulation, Neoplastic , Genes, ras , Nasopharyngeal Neoplasms , Genetics , Metabolism , Pathology , Plasmids , Proto-Oncogene Proteins p21(ras) , Metabolism , RNA Interference , RNA, Small Interfering , Genetics , Transfection
15.
Journal of Southern Medical University ; (12): 1415-1417, 2010.
Article in Chinese | WPRIM | ID: wpr-336164

ABSTRACT

<p><b>OBJECTIVE</b>To explore the correlation of ras gene to the drug resistance of nasopharyngeal carcinoma to cetuximab.</p><p><b>METHODS</b>Cultured 5-8F/Erbitux cells were induced by stepwise exposure to increasing doses of cetuximab. MTT assay was used to determine the IC50 (half inhibitory concentration) of cetuximab and the drug resistance index (RI). Western blotting was employed to detect the protein levels of H-ras and K-ras. Real-time PCR was used to detect the expression of H-ras and K-ras. Gene sequencing was performed to identify potential mutations in H-ras and K-ras genes.</p><p><b>RESULTS</b>We successfully induced cetuximab-resistant 5-8F/Erbitux hNPC cells by stepwise exposure to increasing doses of cetuximab. After treatment with cetuximab for 3 and 5 days, the RI of 5-8F/Erbitux cells was 1.2 and 1.1, respectively. The 5-8F/Erbitux cells had increased levels of H-ras and K-ras protein expressions (P<0.001) and enhanced gene expressions of H-ras (P=0.016) and ras-p21 (P=0.113) with decreased K-ras gene expression (P=0.000). Sequence analysis identified no mutations in the H-ras and K-ras genes in codons 12, 13, 59, and 61.</p><p><b>CONCLUSION</b>Gene amplification and overexpression of H-ras is the major mechanism that causes the drug resistance of 5-8F/Erbitux cells to cetuximab.</p>


Subject(s)
Humans , Antibodies, Monoclonal , Pharmacology , Antibodies, Monoclonal, Humanized , Cell Line, Tumor , Cetuximab , Drug Resistance, Neoplasm , Genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Genes, ras , Genetics , Nasopharyngeal Neoplasms , Genetics , Pathology
16.
Chinese Journal of Oncology ; (12): 575-579, 2010.
Article in Chinese | WPRIM | ID: wpr-293513

ABSTRACT

<p><b>OBJECTIVE</b>To establish a cetuximab-resistant human nasopharyngeal carcinoma 5-8F/Erbitux cell line and preliminarily study the relationship between the insulin-like growth factor 1 receptor (IGF-1R) signaling pathway and the resistance of nasopharyngeal carcinoma to cetuximab.</p><p><b>METHODS</b>A nasopharyngeal cancer cell line, 5-8F, with high epidermal growth factor receptor (EGFR) expression and cetuximab sensitivity, was selected as study object. The cetuximab-resistant 5-8F/Erbitux cell line was induced by stepwise selection after exposure to increasing doses of cetuximab. The IC(50) was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the resistance index (RI) was calculated. The growth curves of 5-8F and 5-8F/Erbitux cells were plotted and the doubling times were calculated by cell counting assay. The cell cycle was detected by flow cytometry. Cross-resistance profiles of 5-8F/Erbitux cells to 5-Fu, Taxol and DDP were tested by MTT assay. Expression levels of P-gP, IGF-1R and P-IGF-1R of 5-8F and 5-8F/Erbitux cells were determined by Western blot analysis and MDR1 gene by real-time fluorescent quantitative PCR.</p><p><b>RESULTS</b>A cetuximab-resistant human nasopharyngeal carcinoma cell line 5-8F/Erbitux was successfully established and their resistance index (RI) were 1.2 and 1.1, respectively, at 3 d and 5 d of the cetuximab treatment. The doubling times of 5-8F and 5-8F/Erbitux cells were 26.63 h and 142.30 h, respectively. Flow cytometry demonstrated that 5-8F/Erbitux cells showed an increased population at G(0)/G(1) phase (P < 0.001) and reduced population at S phase (P < 0.001), compared with 5-8F cells. The 5-8F/Erbitux cells showed cross-resistance to 5-Fu (RI = 3.95, P < 0.01) and some resistance to Taxol as well as enhanced sensitivity to DDP (P > 0.05 for all). The 5-8F/Erbitux cells also had increased levels of P-gP, IGF-1R and P-IGF-1R compared with 5-8F cells (P < 0.001 for all). Expression of MDR1 gene was not detected in 5-8F cells and only very weak expression in 5-8F/Erbitux cells.</p><p><b>CONCLUSION</b>Cetuximab-resistant 5-8F/Erbitux cells have no common multidrug resistance like that induced by traditional chemotherapeutic drugs. The excessive activation of the IGF-1R signaling pathway is probably one of the mechanisms that caused resistance of 5-8F/Erbitux cells to cetuximab.</p>


Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Metabolism , Antibodies, Monoclonal , Pharmacology , Antibodies, Monoclonal, Humanized , Antimetabolites, Antineoplastic , Pharmacology , Antineoplastic Agents , Pharmacology , Antineoplastic Agents, Phytogenic , Pharmacology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cetuximab , Cisplatin , Pharmacology , Drug Resistance, Neoplasm , Fluorouracil , Pharmacology , Nasopharyngeal Neoplasms , Metabolism , Pathology , Paclitaxel , Pharmacology , Phosphorylation , Receptor, IGF Type 1 , Metabolism , Signal Transduction
17.
Journal of Central South University(Medical Sciences) ; (12): 103-109, 2008.
Article in Chinese | WPRIM | ID: wpr-814113

ABSTRACT

OBJECTIVE@#To investigate the effect of overexpression of glycosylphosphatidyl-inositol-specific phospholipase D (GPI-PLD) on the biological character of hepatocellular carcinoma cell line HepG2.@*METHODS@#The GPI-PLD gene eukaryon expression vector pcDNA3.1(+)/ GPI-PLD was transiently transfected into HepG2 cell by lipid-media transfection. The untransfected HepG2 and HepG2 transfected with pcDNA3.1(+) were used as controls. After screening with G418, the single clone was obtained. The expression level of GPI-PLD mRNA in HepG2 was identified by reverse transcription polymerase chain reaction (RT-PCR). GPI-PLD activities were analyzed quantitatively by triton-X-114 partition with GPI anchored placental alkaline phosphatase (PLAP) as a substrate. Cell count was used to detect the proliferation of the 3 groups, and complement dependent cytotoxicity (CDC) effects were observed by the staining of trypan blue. Apoptosis cells were analyzed by flow cytometry. Carcinoembryonic antigen (CEA)was detected by enzyme linked immunosorbent assay (ELISA).@*RESULTS@#Compared with HepG2 and pcDNA3.1(+)/HepG2 cell, the levels of GPI-PLD activities and its mRNA from pcDNA3.1(+)/GPI-PLD/HepG2 were increased with almost 2 to 5 times,respectively. The GPI anchored PLAP and CEA released into the medium by GPI-PLD, and the rate of CDC killing on the cells were significantly increased. However, the proliferative capacity was obviously decreased, and the typical apoptosis cells were presented in positive clones and its apoptosis rates were increased significantly.@*CONCLUSION@#The stable cell line with overexpression of GPI-PLD has been constructed. The overexpression of GPI-PLD in these cells increases the sensitivity of these cells to CDC killing and impairs the proliferative capacity of cells, and promotes the apoptosis.


Subject(s)
Humans , Apoptosis , Genetics , Carcinoma, Hepatocellular , Genetics , Pathology , Complement Activation , Genetics , Cytotoxicity, Immunologic , Genetics , Eukaryotic Cells , Metabolism , Genetic Vectors , Genetics , Metabolism , Liver Neoplasms , Genetics , Pathology , Phospholipase D , Genetics , RNA, Messenger , Genetics , Transfection , Tumor Cells, Cultured , Up-Regulation
18.
Journal of Southern Medical University ; (12): 858-859, 2006.
Article in Chinese | WPRIM | ID: wpr-282896

ABSTRACT

<p><b>OBJECTIVE</b>To determine the cut-off value of serum neuron-specific enolase (NSE) level for distinguishing small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Serum NSE levels were measured by enzyme-linked immunosorbent assay in 137 patients with NSCLC or SCLC, and the best cut-off value was analyzed using ROC curve.</p><p><b>RESULTS</b>The positivity rate of serum NSE was significantly higher in patients with SCLC than in those with NSCLC (P<0.01). The best cut-off value was 15.45 microg/L using ROC curve, which gave a sensitivity of 66.7% and specificity of 65.7%.</p><p><b>CONCLUSION</b>Serum NSE level may allow simple and cost-effective differentiation of SCLC and NSCLC.</p>


Subject(s)
Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Blood , Carcinoma, Non-Small-Cell Lung , Pathology , Carcinoma, Small Cell , Pathology , Diagnosis, Differential , Lung Neoplasms , Pathology , Phosphopyruvate Hydratase , Blood
19.
Chinese Journal of Lung Cancer ; (12): 154-157, 2004.
Article in Chinese | WPRIM | ID: wpr-345824

ABSTRACT

<p><b>BACKGROUND</b>To evaluate the diagnostic values of multiple tumor marker protein biochip detective system for lung cancer.</p><p><b>METHODS</b>The serum levels of 12 tumor markers, including CA199, NSE, CEA, CA242, CA125, CA153, AFP, ferritin, free-PSA, PSA, β-HCG and HGH, were measured in 108 lung cancer patients, 48 patients with benign pulmonary lesion and 145 healthy by the detective system.</p><p><b>RESULTS</b>The positive rates were 83.33% (90/108), 52.08% (25/48) and 28.97% (42/145) in lung cancer, benign pulmonary lesion and healthy groups, respectively. The lung cancer group had significantly higher positive rate than that of the controls (Chi-Square=16.75 and 73.32, both P < 0.001); There was significant difference of positive rate in various clinical stages of lung cancer (Chi-Square=7.89, P=0.048), but not in different pathologic classification. Serum CA199, CEA and CA242 levels were closely correlated with clinical staging (F=2.84, P=0.041; F= 3.49, P=0.018; F =5.22, P=0.002). The positive rate of CEA in adenocarcinoma was higher, but no significant difference was observed (Chi-Square=0.71, P=0.07). NSE in small cell lung cancer had the highest positive rate (Chi-Square=19.03, P < 0.001). Combined measurement of the twelve markers had higher sensitivity (Chi-Square= 368.58, P < 0.001), but less specificity (Chi-Square= 369.87, P < 0.001).</p><p><b>CONCLUSIONS</b>Combined measurement of various serum tumor markers using protein biochip can significantly increase the diagnostic sensitivity for lung cancer. Meanwhile, it is also significant for defining clinical stage, identificating pathologic classification, as well as monitoring therapeutic efficacy. As its specificity and positive predictive value are lower, it is more suitable to be used as a surveying tool for symptomless people, especially for high risk people for lung cancer.</p>

20.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-557001

ABSTRACT

Objective To compare two assay methods, namely immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), in the detection of Her-2/neu status in mammary cancer tissues. Methods In 20 samples IHC technique was applied to detect the Her-2 protein in the cytomembrane of mammary cancer, and FISH was used to assess the amplification of Her-2/neu gene, and SPSS 10.0 software was employed to analyze the relationship between the two methods. Results The coincident rate of two methods was 85%, therefore there was a significant positive correlation between the two techniques (?~2=80.00, P=0.00). Conclusion The two assay methods, IHC and FISH showed a good coincidence in the detection of Her-2/neu status in mammary cancer tissues.

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