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BACKGROUND@#Emerging evidence indicates that the sineoculis homeobox homolog 1-eyes absent homolog 1 (SIX1-EYA1) transcriptional complex significantly contributes to the pathogenesis of multiple cancers by mediating the expression of genes involved in different biological processes, such as cell-cycle progression and metastasis. However, the roles of the SIX1-EYA1 transcriptional complex and its targets in colorectal cancer (CRC) are still being investigated. This study aimed to investigate the roles of SIX1-EYA1 in the pathogenesis of CRC, to screen inhibitors disrupting the SIX1-EYA1 interaction and to evaluate the efficiency of small molecules in the inhibition of CRC cell growth.@*METHODS@#Real-time quantitative polymerase chain reaction and western blotting were performed to examine gene and protein levels in CRC cells and clinical tissues (collected from CRC patients who underwent surgery in the Department of Integrated Traditional and Western Medicine, West China Hospital of Sichuan University, between 2016 and 2018, n = 24). In vivo immunoprecipitation and in vitro pulldown assays were carried out to determine SIX1-EYA1 interaction. Cell proliferation, cell survival, and cell invasion were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, clonogenic assay, and Boyden chamber assay, respectively. The Amplified Luminescent Proximity Homogeneous Assay Screen (AlphaScreen) method was used to obtain small molecules that specifically disrupted SIX1-EYA1 interaction. CRC cells harboring different levels of SIX1/EYA1 were injected into nude mice to establish tumor xenografts, and small molecules were also injected into mice to evaluate their efficiency to inhibit tumor growth.@*RESULTS@#Both SIX1 and EYA1 were overexpressed in CRC cancerous tissues (for SIX1, 7.47 ± 3.54 vs.1.88 ± 0.35, t = 4.92, P = 0.008; for EYA1, 7.61 ± 2.03 vs. 2.22 ± 0.45, t = 6.73, P = 0.005). The SIX1/EYA1 complex could mediate the expression of two important genes including cyclin A1 (CCNA1) and transforming growth factor beta 1 (TGFB1) by binding to the myocyte enhancer factor 3 consensus. Knockdown of both SIX1 and EYA1 could decrease cell proliferation, cell invasion, tumor growth, and in vivo tumor growth (all P < 0.01). Two small molecules, NSC0191 and NSC0933, were obtained using AlphaScreen and they could significantly inhibit the SIX1-EYA1 interaction with a half-maximal inhibitory concentration (IC50) of 12.60 ± 1.15 μmol/L and 83.43 ± 7.24 μmol/L, respectively. Administration of these two compounds could significantly repress the expression of CCNA1 and TGFB1 and inhibit the growth of CRC cells in vitro and in vivo.@*CONCLUSIONS@#Overexpression of the SIX1/EYA1 complex transactivated the expression of CCNA1 and TGFB1, causing the pathogenesis of CRC. Pharmacological inhibition of the SIX1-EYA1 interaction with NSC0191 and NSC0933 significantly inhibited CRC cell growth by affecting cell-cycle progression and metastasis.
Subject(s)
Animals , Humans , Mice , Cell Line, Tumor , Cell Proliferation , Colorectal Neoplasms/genetics , Gene Expression Regulation, Neoplastic , Genes, Homeobox , Homeodomain Proteins/metabolism , Intracellular Signaling Peptides and Proteins , Mice, Nude , Nuclear Proteins/genetics , Protein Tyrosine Phosphatases/geneticsABSTRACT
Objective:To investigate the effect of virtual reality balance training on knee proprioception after anterior cruciate ligament reconstruction (ACLR). Methods:From January, 2016 to December, 2018, 72 patients after ACLR were randomly divided into control group (n = 36) and observation group (n = 36). On the basis of routine rehabilitation, they received routine balance training and balance training with virtual reality, respectively, for eight weeks. Before and after training, the center of pressure (COP) time stability, the COP speed (COPS) of closed eyes, the COPS of one leg, the knee joint reaction time, the center of pressure area (COPA) and the active joint angle recurrence deviation were measured. Results:After training, the COP time stability, the COPS of closed eyes, the COPS of one leg, knee joint reaction time, COPA and active joint angle recurrence deviation improved in both groups (|t|>5.611, P < 0.001); and the COP time stability, the COPS of closed eyes, the COPS of one leg, the knee joint reaction time and active joint angle recurrence deviation were better in the observation group than in the control group (|t| > 3.232, P < 0.01), however, no significant difference was found in COPA between two groups (t = 1.054, P = 0.296). Conclusion:Virtual reality balance training can improve the proprioception of knee joint after ACLR, which is better than routine balance training.
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BACKGROUND@#Previously, the authors modified the surgical technique to preserve tibial bone mass for Oxford unicompartmental knee arthroplasty (UKA). The purpose of this study was to determine the clinical outcomes and values of this modified technique.@*METHODS@#Clinical data of 34 consecutive patients who underwent the unilateral modified UKA technique (modified group, 34 knees) were retrospectively analyzed. To compare the outcome, a match-paired control group (conventional group, 34 knees) of an equal number of patients using the conventional technique system in the same period were selected and matched with respect to diagnosis, age, pre-operative range of motion (ROM), and radiological grade of knee arthrosis. Clinical outcomes including knee Hospital for Special Surgery (HSS) score, ROM, and complications were compared between the two groups. Post-operative radiographic assessments included hip-knee-ankle angle (HKA), joint line change, implant position, and alignment.@*RESULTS@#The mean follow-up time was 38.2 ± 6.3 months. There was no difference in baseline between the two groups. The amount of proximal tibial bone cut in the modified group was significantly less than that of the conventional group (4.7 ± 1.1 mm vs. 6.7 ± 1.3 mm, t = 6.45, P < 0.001). Joint line was elevated by 2.1 ± 1.0 mm in the modified group compared with -0.5 ± 1.7 mm in the conventional group (t = -7.46, P < 0.001). No significant differences were observed between the two groups after UKA with respect to HSS score, VAS score, ROM, and HKA. Additionally, the accuracy of the post-operative implant position and alignment was similar in both groups. As for implant size, the tibial implant size in the modified group was larger than that in the conventional group (χ = 4.95, P = 0.035).@*CONCLUSIONS@#The modified technique for tibial bone sparing was comparable with the conventional technique in terms of clinical outcomes and radiographic assessments. It can preserve tibial bone mass and achieve a larger cement surface on the tibial side.
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Background@#Previously, the authors modified the surgical technique to preserve tibial bone mass for Oxford unicompartmental knee arthroplasty (UKA). The purpose of this study was to determine the clinical outcomes and values of this modified technique.@*Methods@#Clinical data of 34 consecutive patients who underwent the unilateral modified UKA technique (modified group, 34 knees) were retrospectively analyzed. To compare the outcome, a match-paired control group (conventional group, 34 knees) of an equal number of patients using the conventional technique system in the same period were selected and matched with respect to diagnosis, age, pre-operative range of motion (ROM), and radiological grade of knee arthrosis. Clinical outcomes including knee Hospital for Special Surgery (HSS) score, ROM, and complications were compared between the two groups. Post-operative radiographic assessments included hip-knee-ankle angle (HKA), joint line change, implant position, and alignment.@*Results@#The mean follow-up time was 38.2 ± 6.3 months. There was no difference in baseline between the two groups. The amount of proximal tibial bone cut in the modified group was significantly less than that of the conventional group (4.7 ± 1.1 mm vs. 6.7 ± 1.3 mm, t = 6.45, P < 0.001). Joint line was elevated by 2.1 ± 1.0 mm in the modified group compared with -0.5 ± 1.7 mm in the conventional group (t = -7.46, P < 0.001). No significant differences were observed between the two groups after UKA with respect to HSS score, VAS score, ROM, and HKA. Additionally, the accuracy of the post-operative implant position and alignment was similar in both groups. As for implant size, the tibial implant size in the modified group was larger than that in the conventional group (χ2 = 4.95, P = 0.035).@*Conclusions@#The modified technique for tibial bone sparing was comparable with the conventional technique in terms of clinical outcomes and radiographic assessments. It can preserve tibial bone mass and achieve a larger cement surface on the tibial side.
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Objective To explore the influence factors of the compliance in colorectal cancer( CRC)screening. Methods A total of 140 096 local people were selected for fecal occult blood test( FOBT)in Songjiang District Shanghai City. They were over 40 years old and thought to be high risk groups of colorectal cancer. The participants with FOBT positive were asked to following colonoscopy. All the participants were interviewed by a questionnaire. The influencing factors of compliance on colorectal cancer screening were analyzed by multivariate regression. Results Of all the residents with high risk of colorectal cancer,The FOBT positive rate was 16. 83%. Of the FOBT positive residents,8 566 cases took the examination of colonoscopy,and the screening rate was 36. 33%. A total of 142 cases were diagnosed as colorectal cancer, with the detection rate of 1. 66%. Compared with the positive rate of FOBT in 2012 and 2013,the rate was significantly lower(18. 16%,15. 78%,p<0. 05),and the rate of colonoscopy examination increased(45. 66%,26. 04%,p<0. 05). Multivariate logistic regression showed that fear of pain( OR=3. 01 ),the thought of uselessness( OR=1. 89 )were risk factors for compliance in colorectal cancer screening( p <0. 05 ). Conclusion The compliance of colorectal cancer screening in Songjiang District is relatively low. High-risk groups with regular physical examination,high level education were prone to have good compliance,and fear of pain,think it useless were the main reason for incompliance.
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<p><b>OBJECTIVE</b>To investigate the relationship between p16 expression and cell proliferation and prognosis in gastric cancer patients.</p><p><b>METHODS</b>Gastric cancer cell lines SGC-7901, MKN45, MKN28, human embryonic kidney cell line HEK293, human fibroblast cell line MRC-5, and surgical specimens of gastric carcinoma and adjacent normal gastric mucosa from 65 patients were included in this study. RT-PCR, MTT and FCM assays were used to detect p16 expression in gastric cancer cell lines and surgical specimens of gastric cancer. MTT assay was used to determine cancer cell viability and FCM to detect cell cycle. Kaplan-Meier survival curve and Log-Rank statistics were used to analyze the relationship between p16 expression and survival of petients with gastric cancer.</p><p><b>RESULTS</b>Gastric cancer cell lines were mostly negative for p16 expression, and p16 was re-expressed after the cells transfected with p16 gene by adenovirus AdCMV-p16. p16 re-expression resulted in the decrease of cancer cell viability and cancer cell cycle arrest with increased G(1) phase and decreased S phase. p16 expression in cancer specimens was 32.3% (21/65), significantly lower than the 81.5% (53/65) in normal mucosa (χ(2) = 32.124, P < 0.001). The disease-free survival was significantly shorter in p16-negative patients than that in p16-positive patients (P < 0.01), but not the overall survival (P > 0.05). p16 expression was significantly correlated with differentiation and lymph node metastasis, but not significantly correlated with sex, age, tumor size or invasion depth of the gastric cancer.</p><p><b>CONCLUSIONS</b>p16 gene is important for cancer cell proliferation. The inactivation gives cancer cells a high activity for proliferation and metastasis, and then influences the disease-free survival of gastric cancer patients.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adenoviridae , Genetics , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Cell Survival , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Metabolism , Disease-Free Survival , Gene Expression Regulation, Neoplastic , Genes, p16 , Lymphatic Metastasis , Neoplasm Invasiveness , RNA, Messenger , Metabolism , Recombinant Proteins , Genetics , Metabolism , Stomach Neoplasms , Metabolism , Pathology , Survival Rate , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of different occlusal thickness and shoulder finish line depth on stress distribution of all-ceramic crowns and to select optimal occlusal thickness and shoulder finish line depth using continuous variation of parameters.</p><p><b>METHODS</b>This analysis was performed using mandibular first molar finite element model. The range of occlusal thickness was set from 1 mm to 3 mm, and that of shoulder finish line depth was from 0.4 mm to 1.2 mm. Load of 225 N was applied perpendicularly to the occlusal surface of the tooth at all buccal cusps to simulate functional occlusal force. The maximum equivalent stresses in crown, cement layer, abutment, periodontal ligament, alveolar bone were calculated, and the sensitivities of stresses to the variables were also evaluated.</p><p><b>RESULTS</b>The maximum equivalent stresses in crown, cement layer and abutment decreased as occlusal thickness and shoulder finish line depth were increased, while no obvious change were found in maximum equivalent stresses in periodontal ligament and alveolar bone. When occlusal thickness exceeded 1.87 mm and shoulder finish line depth exceeded 0.66 mm, the tangent slope rate of the maximum equivalent stress response curves ranged from -1 to 1. Data indicated that occlusal thickness played a more important role in reducing maximum equivalent stress in cement layer than finish line depth did, and shoulder finish line depth was a more effective parameter in reducing maximum equivalent stress in crown and abutment than occlusal thickness was.</p><p><b>CONCLUSIONS</b>Occlusal thickness exceeding 1.87 mm and shoulder finish line depth exceeding 0.66 mm are optimal design for ceramic crown on mandibular first molar from biomechanical point of view.</p>
Subject(s)
Humans , Ceramics , Chemistry , Crowns , Dental Cements , Chemistry , Dental Occlusion , Dental Prosthesis Design , Dental Stress Analysis , Finite Element Analysis , Mandible , Molar , Tensile StrengthABSTRACT
<p><b>OBJECTIVE</b>To construct a bait plasmid containing human telomerase RNA with multiple point mutations in a yeast three-hybrid system and evaluate the toxicity of the recombinant bait plasmid.</p><p><b>METHODS</b>The primers were designed according to the hTR sequence and the target mutation sites for inducing T-->A mutations at the 41st, the 80th and 102nd nucleotides of the hTR gene using the overlapping extension PCR (OE-PCR) method. The mutant was cloned into PMD18T vector, confirmed by sequencing, sub-cloned into the bait plasmid PRH3' and identified with PCR and restriction enzyme digestion. The recombinant bait plasmid was then transformed into yeast L40 ura3/pHyblex/ZeoMS2 for toxicity test.</p><p><b>RESULTS</b>Sequence analysis demonstrated successful introduction of point mutations at the target sites without causing random mutation. The recombined bait plasmid constructed showed no obvious toxicity against the host yeast cells.</p><p><b>CONCLUSIONS</b>The recombinant plasmid containing the human telomerase RNA mutant (PRH3'-hTRm) has been successfully constructed and can be used as the bait plasmid in yeast three-hybrid system.</p>
Subject(s)
Humans , Base Sequence , Plasmids , Genetics , Point Mutation , Polymerase Chain Reaction , RNA , Genetics , Telomerase , Genetics , Two-Hybrid System TechniquesABSTRACT
Percutaneous osteoplasty (POP) as a technical extension of percutaneous vertebroplasty (PVP) has been used to treat malignant disease that affects the skeletal system. POP has demonstrated good outcome for pain relief and functional improvement. Few studies have reported on the efficiency of POP to treat malignancies located in the femoral head. We designed a pilot study with the use of POP to treat intractable pain caused by a femoral head metastatic tumor in a 43-year-old man. During the follow-up period, the patient experienced sustained pain relief and improvement of quality of life that persisted for more than three months.
Subject(s)
Adult , Humans , Male , Diagnosis, Differential , Femoral Neoplasms/diagnosis , Lung Neoplasms/pathology , Magnetic Resonance Imaging , Tomography, Emission-Computed , Tomography, X-Ray ComputedABSTRACT
<p><b>OBJECTIVE</b>To develop a novel vector system, which combines the advantages of the gene therapy, antiangiogenic therapy and virus therapy, and to observe its effect on lung cancer.</p><p><b>METHODS</b>Human angiostatin gene hA(k1-5) was inserted into the genome of the replicative virus specific for the tumor cells by virus recombination technology. The expression of hA(k1-5), its effect on tumor growth in vitro and in vivo were studied.</p><p><b>RESULTS</b>A new kind of gene-viral vector system, designated as CNHK200-hA(k1-5), in which the E1b55 000 gene was deleted but the E1a gene of adenovirus preserved, was constructed. The novel vector system possessed the same property as the replicative virus ONYX-015, which replicates in p53- tumor cells but not in normal cells, thus specifically kills tumor cells. In vitro, CNHK200-hA and Ad-hA both could kill A549 tumor cells but the latter needed 100 times more MOI to achieve the same amplitude of cell killing. In vivo, the therapeutic effect of CNHK200-hA on human lung cancer A549 xenograft in nude mice was significantly better than that of Ad-hA and that of tumor-replicative virus ONYX-015.</p><p><b>CONCLUSION</b>CNHK200-hA(k1-5), a novel vector is constructed in which the angiostatin gene is inserted into the genome of the replicative adenovirus cytotoxic to p53-negative tumor cells. It has the advantages of specific tumor targeting, high level gene expression in tumor cells, and potent tumoricidal activity.</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Adenoviridae , Genetics , Adenovirus E1A Proteins , Genetics , Angiostatins , Genetics , Physiology , Cell Line, Tumor , Cell Survival , Genetic Therapy , Genetic Vectors , Lung Neoplasms , Metabolism , Pathology , Therapeutics , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To observe the injury of plumbism to the nerve system in children.</p><p><b>METHODS</b>One hundred children with plumbism which had no masculine body sign of nerve system were divided into two groups. Fifty of them were as controls, while others were treated by calcium. Fifty age and gender matched healthy children were selected as the normal group in order to compare with plumbism group. All of their auditory brainstem response (ABR), pure tone and distortion product otoacoustic emission (DPOAE) were examined.</p><p><b>RESULTS</b>Latency period of wave V of the plumbism groups was longer than that of normal group. The difference between them was significant statistically (t =3.21, P <0.01). The interval between wave I and wave III of plumbism group was longer than that of normal group, but there was no difference between them (former: t = 2.06, P > 0.05; later: t = 1.64, P > 0.05). The interval between wave III and wave V and the interval between wave I and wave V of plumbism group was longer than that of normal group, and their difference was significant statistically (former: t = 7.30, P <0.01; later: t = 3.14, P < 0.01). The interval between wave I and wave III of plumbism group was longer than that of normal group, but there was no difference between them (t = 0.91, P > 0.05). The amplitude of wave I, wave III and wave V of plumbism group was lower than that of normal group, and their difference was significant statistically (I: t =3.20, P < 0.01; III: t=3.31, P < 0.01; V: t=3.41, P < 0.01). The auditory brainstem response, pure tone and DPOAE in all three groups had no obvious change after six months treatment (P >0.05).</p><p><b>CONCLUSIONS</b>Plumbism may have potential clinical injury to nerve system. There has no obvious change of ABR after treatment by calcium. ABR may be a forepart examination to find the injury of plumbism to the nerve system.</p>
Subject(s)
Child , Humans , Case-Control Studies , Evoked Potentials, Auditory, Brain Stem , Lead PoisoningABSTRACT
<p><b>OBJECTIVE</b>To evaluate the therapeutic effect and the expression level of a tumor-specific replication-competent adenovirus and a replication-defective adenovirus expression mouse recombinant IL-12 (mIL-12) gene on hepatocellular carcinoma (HCC) in vitro.</p><p><b>METHODS</b>The cytotoxicity of replication-competent adenovirus with E1B-55 000 attenuated CNHK200-mIL12 and ONYX-015 (dl1520), and replication-defective adenovirus Adv-mIL12 were evaluated by MTT and replication assay in two HCC cell lines (HepG2 and Hep3B) and human normal hepatocyte line (LO2). Western blot and ELISA were used to determine the expression level of mIL-12.</p><p><b>RESULTS</b>CNHK200-mIL12 replicated in HepG2 and Hep3B with an increase of 3,160-fold and 630-fold respectively in 96 h post-infection. CNHK200-mIL12 could kill HepG2 and Hep3B cells at a very low MOI (Multiplicity of Infection) and in short time course (HepG2:MOI = 0.2, on day 4; Hep3B:MOI = 0.005, on day 2), while it had no significant effect on LO2. Furthermore, the expressing level of mIL-12 in CNHK200-mIL12 treated HCC cell lines was much higher than that in Adv-mIL12 treated one (HepG2 101-fold, Hep3B 20-fold respectively).</p><p><b>CONCLUSION</b>Replication-competent adenovirus is more effective than replication-defective adenovirus in both cytotoxicity and efficiency of gene transfer in HCC, and holds great promise in the area of HCC therapy.</p>