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Objective:To discuss the effect of downregulating of high mobility group box protein 2(HMGB2)expression on the biological behavior of the liver cancer cells and the epithelial-mesenchymal transition(EMT)process,and to clarify its mechanism.Methods:The human liver cancer LM3 cells at logarithmic growth phase were divided into negative control group and HMGB2 RNA interference group(HMGB2 siRNA group);the cells in two groups were transfected with RNA oligonucleotides(RNA oligos)with irrelevant sequences and RNA oligos designed to knock down HMGB2,and the Lipofectamine 2000 was regarded as the vector.The expression levels of HMGB2 mRNA and protein in the cells in two groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods;cell scratch assay and Transwell chamber assay were used to detect the migration and invasion abilities of the cells in two groups;the expression levels of E-cadherin,N-cadherin,and Vimentin proteins and protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway related proteins in the cells in two groups were detected by Western blotting method.Results:Compared with negative control group,the expression levels of HMGB2 mRNA and protein in the cells in HMGB2 siRNA group were significantly decreased(P<0.05),the cell scratch healing rate was significantly decreased(P<0.01),the number of invasion cells was significantly decreased(P<0.01),and the expression level of E-cadherin protein in the cells was significantly increased(P<0.01),while the expression levels of N-cadherin,Vimentin,mTOR,AKT,and phosphorylated AKT(p-AKT)proteins in the cells were significantly decreased(P<0.05 or P<0.01).Conclusion:Downregulating the expression of HMGB2 can reduce the migration and invasion abilities of the liver cancer LM3 cells and inhibit the EMT,and its mechanism may be related to regulating the expression of the AKT/mTOR pathway related proteins.
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The adjacent anatomy of the pelvis is complicated, with digestive, urinary, reproductive and other organs as well as important blood vessels and nerves. Therefore, accurate resection of pelvic tumors and precise reconstruction of defects after resection are extremely difficult. The development of medical 3D printing technology provides new ideas for precise resection and personalized reconstruction of pelvic tumors. The “triune” application of 3D printing personalized lesion model, osteotomy guide plate and reconstruction prosthesis in pelvic tumor limb salvage reconstruction treatment has achieved good clinical results. However, the current lack of normative guidance standards such as preparation and application of 3D printing personalized lesion model, osteotomy guide plate and reconstruction prosthesis restricts its promotion and application. The formulation of this consensus provides normative guidance for 3D printing personalized pelvic tumor limb salvage reconstruction treatment.
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Objective:To evaluate the clinical efficacy of staged surgery in treatment of calf Gustilo-Anderson type IIIC fracture.Methods:A retrospective case series was conducted to analyze clinical data of 16 patients with calf Gustilo-Anderson type IIIC fracture admitted to Shanghai Jiao Tong University Affiliated Sixth People's Hospital from January 2017 to December 2019. There were 12 males and 4 females, with the age of (38.6±8.2)years (range, 18-53 years). All patients had limb salvage treatment at one stage in the emergency department. The survival of the limb and the occurrence of vascular crisis were examined within one week after limb salvage. The second stage involved the repair of skin and soft tissue defects with the defect area from 12.0 cm×5.0 cm to 20.0 cm×8.0 cm using free flaps. The survival of the flap, vascular crisis, and donor site healing within two weeks after the flap procedure. The third stage used bone graft revision and bone lengthening technology to repair bone tissue. The lower extremity functional scale (LEFS) and Mazur ankle joint function score were used to evaluate the function of the affected limb before bone repair and at the last follow-up. The fracture healing and related complications were observed at the last follow-up.Results:All patients were followed up for (14.2±4.6)months (range, 8-20 months). At one stage, the limb-saving surgery was successful in all patients, among which one had vascular crisis. At second stage, free flaps survived in all patients, among which two had vascular crisis. All donor areas were healed by first intention. At third stage, the LEFS of the affected limb was increased from (32.0±7.4)points before bone repair to (48.0±10.2)points at the last follow-up ( P<0.01) and the Mazur score was increased from (50.9±15.3)points before bone repair to (73.8±11.9)points at the last follow-up ( P<0.01). All bone defects were repaired and healed without complications such as infection or osteomyelitis at the last follow-up. Conclusion:For calf Gustilo-Anderson type IIIC fracture, the staged strategy can effectively save limbs and restore limb function.
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Objective:To propose and verify a surgical classification system for the axial primary malignant and aggressive benign tumor.Methods:The CZH surgical classification system was originally developed for the axial primary malignant and aggressive benign tumor. The CZH surgical classification system includes seven types, according to the anatomic features and the extension of tumor violation. A total of 136 patients (79 males and 57 females) with axial primary malignant and aggressive benign tumor from multiple tertiary centers who received surgery from July 2006 to July 2019 were included. The average age was 44.40±17.55 years (8-83 years) old. There were 99 malignant tumors and 37 aggressive benign tumors included. The number of patients with each classification was presented as followed, Type I 13, Type II 15, Type IIIa 3, Type IIIb 20, Type IVa 43, Type IVb 12, Type Va 21, Type Vb 3, Type VI 2, Type VIIa 3 and Type VIIb 1. Surgical procedures were selected according to different types in classification. The inter- and intra-observer consistencies were evaluated by the Kendall's W test. The VAS, Frankel score, overall survival and recurrence free survival were recorded during the follow-up. Results:The inter- and intra-observer consistent coefficient was 0.973 and 0.996, respectively ( P<0.05). The single posterior approach was adopted for the Type II tumors. Other patients underwent surgery by the combined antero-posterior approach. The majority in anterior approach (113 cases) was the modified submandibular approach. The reconstruction modes included anterior "T" shape titanium mesh (112 cases) or the 3D printed prothesis (7 cases) combined with the posterior occipto-cervical fusion (92 cases) or the pedicle screw system (44 cases). The average surgical duration and the volume of intraoperative bleeding was 348.40±136.14 min (60-760 min) and 1 225.69±859.40 ml (80-4 000 ml), respectively. The operation duration and volume of intraoperative bleeding among each type were with statistical difference. The patients with Type IV, V tumors had longer operation duration than those with Type II tumors. Those with Type V and VII tumors had longer operation duration than those with Type I tumors. The patients with Type V tumors had more intraoperative bleeding than those with Type I-IV tumors. The average preoperative VAS score was 4.15±2.25 and then was reduced significantly to 0.62±0.71 and 0.38±0.59 at one and three months after operation, respectively. The Frankel score was also significantly ameliorated at one and three months postoperatively. There were 22 postoperative complications (16.2%). The complications included cerebral spinal fluid leak (12.5%), dysphagia and/or dysphonia (7.4%), dyspnea (5.1%), wound infection (3.7%), wound hemorrhage (2.2%) and pharyngeal dehiscence (1.5%). The incidence of postoperative complication was 25.9% in Type IV-VII tumors, while 11.8% in Type I-III tumors. Conclusion:CZH surgical classification system was verified with high observer consistency. This classification system could assist surgeons to select proper surgical approaches, resection modes and reconstruction modes, and thus ensure the safety of surgery and reduce the recurrence. The tumors in Type IV, V and VII may be with more challenging for surgeons. The incidence of postoperative complication in Type IV-VII tumors may be higher than that in Type I-III tumors.
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Objective To study the feasibility and safety of total en bloc spondylectomy (TES) for bone tumors of the fourth lumbar spine and evaluate the clinical outcomes.Methods From March 2011 to December 2013,21 patients undergone total en bloc spondylectomy in posterior-only approach were retrospectively reviewed.The patients included 9 males and 12 females,with a mean age of 47.1± 15.6 years old (range,15-71 years old).This series included 12 cases of primary bone tumors and 9 cases of solitary metastases.Preoperative evaluation according to clinical,imaging and pathologic features was performed meticulously to select patients.The length of surgery,estimated blood loss,surgical margins,instrumentation failure,perioperative complications,Frankel scale,visual analogue scale (VAS) for pain,local control rate and overall survival were reviewed and analyzed.Results Total en bloc spondylectomy was performed successfully in all patients.Average operative time and estimated blood loss were 297.6±44.6 min (range,225-420 min) and 2 247.1±904.5 ml (range,900-4100 ml),respectively.The mean follow-up time was 50.4± 17.1 mons (range,24-79 mons).All patients encountered nerve roots stretch and 5 patients (23.8%) showed lower extremeties neurological dysfunction.All of them improved in 2-4 weeks postoperatively and recovered completely at 6-month follow -up.Cerebrospinal fluid leak was found in 4 patients (19.0%).The VAS score was 1.5±1.4 at post-operation,which was significantly lower than the 6.2± 1.6 in average at operation (P=0.008).Three patients with metastatic tumors died during the follow-up.Titanium mesh cage subsidence was observed in 7 patients (33.3%).No implant failure was occurred during the follow-up.Conclusion Total en bloc spondylectomy for tumors of the fourth lumbar spine in a posterior-only approach is feasible.However,there are many intraoperative neurological complications and the indications for TES are extremely limited.
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Objective:To investigate the effects of different doses and infusion methods of recombinant mouse interleukin-18(rmIL-18) on the survival time and tumor diameter of the mice with hepatocellular carcinoma,and to elucidate the rational application of rmIL-18 in vivo.Methods:A total of 60 Babl/C mice were randomly divided into 5 μg rmIL-18 intraperitoneal injection group,0.5 μg rmIL-18 intraperitoneal injection group,0.5 μg rmIL-18 tumor injection group,cytotoxic T lymphocyte(CTL) intraperitoneal injection group,CTL tumor injection group and saline control group;there were 10 mice in each group.From the 10th day of inoculation,the mice in different rmIL-18 groups were injected with the corresponding doses and methods.The mice in different CTL groups were injected with tumor-specific CTL (1×106/mouse) by intraperitoneal and intratumoral injection.The mice in saline control group were injected with an equal volume (100 μL) of saline,the injections were performed 10 times.The diameters of mice were measured weekly and the survival time was recorded.Results:Compared with 5 μg rmIL-18 intraperitoneal injection group,0.5 μg rmIL-18 intraperitoneal injection group and saline control group,the tumor growth rate of the mice in 0.5 μg rmIL-18 tumor injection group was decreased (P<0.01)and the survival rate of the mice was increased (P<0.01);compared with 0.5 μg rmIL-18 intraperitoneal injection group,the tumor growth rate and the survival rate of the mice in CTL intraperitoneal injection group were decreased (P<0.01);compared with 0.5 μg rmIL-18 tumor injection group,the tumor growth rate and the survival rate of the mice in CTL tumor injection group were decreased (P<0.01).Conclusion:The best way for rmIL-18 anti-tumor effect is tumor injection and the effect has a dose-dependent manner.
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Objective To investigate the safety and feasibility of en-bloc resection of a primary sacral chordona based on a 3-dimensional printing model.Methods 31 patients with primary sacral chordoma underwent en-bloc resection via a onestage posterior approach or combined anterior and posterior approaches in our oncology department from January 2013 to December 2014.They comprised 21 males and 10 females of mean age (49.2±12.5) years (range,26-67 years).Preoperative 3-D printing models were created by 3D printing technology,it included tumor tissue,the around vascular and nerves involved in sacral chordoma.The sacral chordomas were en-bloc resection with decompression and internal fixation.Results With the mean (29.0±6.8)months follow-up (range from 19 to 41),all patients underwent en bloc excision via 26 cases with posterior approach,5 cases combined posterior and anterior approaches in one stage.The mean operative time and estimated blood loss were (275.0±58.1) min and (3 250.0±1 304.4) ml,respectively.The visual analogue scale (VAS) score was (5.6±1.9) in average (range from 3 to 9) at preoperation,and (2.0±1.5) at post-operation,which was significantly lower than that of preoperation,and the pain was relief obviously.There were 13 cases in grade C,11 cases in grade D,7 cases in grade E of American Spinal Injury Association (ASIA) grade neurological function before surgery,compared with the pre-operation,there were 5 cases in grade C,6 cases in grade D,20 cases in grade E of post-operation,which was significantly improved.MSTS (Musculoskeletal Tumor Society) 93 score was 6-29 points (20.0%-96.7%) at the follow-up 3 months after surgery,with the average of (19.8 ± 5.8) points,which excellent in 8 cases,good in 14 cases,general in 5 cases,poor in 4 cases.Two cases of dysporia for the reasons of resecting on one side of the S1,2 nerve roots involved by the sacral chordoma,after sacrificing the nerve root of complete tumor resection,the urine left dysfunctional,while the pain of other 29 patients were thoroughly relief after surgery.The ones were relieved with the disturbance of sensation of the perineum before the operation.2 cases were recovery of leakage of cerebrospinal by the drainage of lumbar cistern with normal temperature.One hypostatic pneumonia patient was cured by anti-inflammatory.One with the urinary infection got better by the effective bladder irrigation,which had diabetics mellitus with the bladder stoma before.1 case of skin necrosis due to vascular thrombosis before operation,recevied flap translocation half month after surgery,got recovery 3 months later.Only one underwent tumor resection for the recurrence at 15 months follow-up.Conclusion It is feasible and safe to perform en bloc resection of primary sacral chordoma.This is the most effective means of managing method of the marginal resection of the tumor.Preoperative 3-D printing modeling enables better anatomical understanding of the relationship between the tumor,and can avoid vascular and nerves tissue injury,which can also assist in planning the surgical procedure,and be worth recommendation.
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Objective:To observe the specific killing effect on tumor cells of the spleen cells in mice immunized with three tandem repeats of CEA minigene DNA vaccine pcDNA-triCEA625-667 and to evaluate the safety of the vaccine. Methods: The BALB/c mice were randomly divided into blank vector group ( pcDNA3. 0 ) , haploid vaccine group ( pcDNA-CEA625-667 ) and tandem repeats vaccine group (pcDNA-triCEA625-667). The mice received a total of 4 intramuscular immunization every 10 days once. The changes of body weight,survival state were recorded and the levels of serum ALT and serum creatinine were detected. The specific CTL killing activity of spleen cells in accinated mice on mouse hepatoma cells(H22-CEA+),gastric cancer cells(MFC-CEA+),colorectal cancer cells ( CT26-CEA+) with high expression of CEA and mouse hepatoma cells ( H22-CEA-) without expression of CEA was detected. Results:The two vaccines had strong killing activity on CEA positive liver cancer,gastric cancer and colon cancer cells,and the difference was statistically significant ( P<0. 01 ) compared with the PcDNA3. 0 group. And they had almost no effect on CEA negative tumor cells (H22-CEA-). The killing activity on liver cancer cell(H22-CEA+) and gastric cancer cell(MFC-CEA+) induced by pcDNA-triCEA625-667 was stronger than that induced by pcDNA-triCEA625-667(P<0. 05). The survival status,change of body weight and function of liver and kidney of the mice were not affected by the vaccine. Conclusion:There was no adverse reaction in the course of vaccine immunization. The minigene DNA vaccine derived from CEA can induce tumor specific CTL effect and the immune response level elicited by three tandem repeats of minigene DNA vaccine was superior to that elicited by haploid vaccine.
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Objective:To observe the inhibitory effect of haploid vaccine pcDNA-CEA625-667 and three tandem repeats of minigene DNA vaccine pcDNA-triCEA625-667 derived from CEA gene on tumor in mice bearing tumor and the changes of survival time. Methods:The experimental animal model of mouse liver cell carcinoma was established and the mice were immunized with pcDNA-CEA625-667 and three series of DNA vaccine. Some of the mice were treated with normal saline as control group. The growth curve of tumor growth curve was recorded and the effect of vaccine on the survival time of tumor bearing mice was observed. Results:Compared with the normal saline control group,the two vaccines were able to significantly inhibit the tumor size and growth rate ( P<0. 01 ) of CEA positive tumor bearing mice,the inhibition of pcDNA-triCEA625-667 vaccine group was significantly better than the pcDNA-CEA625-667 vaccine group (P<0. 01),while the two were not inhibited tumor growth in CEA negative tumor bearing mice. The average survival time of the pcDNA-CEA625-667 vaccine group was(48. 50±6. 73)d,and there was significant difference (P<0. 01) compared with the saline control group ( 39. 00 ± 6. 64 ) d. The survival time ( 48. 50 ± 6. 73 ) d of the pcDNA-triCEA625-667 vaccine group was significantly higher than that of the normal saline control group and the pcDNA-CEA625-667 vaccine group (P<0. 01). The survival time of CEA negative tumor bearing mice could not be prolonged in the two groups. Conclusion:Either the haploid or the three series of the DNA vaccine,were able to significantly inhibit tumor growth rate (P<0. 01) and significantly prolong the survival time (P<0. 01) of CEA positive tumor bearing mice,but they had no therapeutic effect on CEA negative tumor bearing mice.
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Objective:To observe the immunological activity of haploid vaccine and three tandem repeats of minigene DNA vac -cine derived from Carcinoembryonic Antigen (CEA) gene.Methods:The immunoreaction was induced by intramuscular injection with pc-DNA3.0,pcDNA-CEA625-667 and pcDNA-triCEA625-667 in BALB/c.Four weeks after injection,the spleen cells and serum were separa-ted respectively from the mice for the in vitro assessment .Changes of the T lymphocytes subset was analyzed by flow cytometry .Lymph proliferation responses were tested by 3 H-TdR incorporation ,IFN-γ,IL-4 and GM-CSF in their cultural supernatants were detected with ELISA and seral IgG antibody against CEA were detected with Western blot and ELISA .Results:The difference of the ratio of CD 4+/CD8+of the mice immuned by pc-DNA3.0,pcDNA-CEA625-667 or pcDNA-triCEA625-667 was not significant.Lymph proliferation responses were more significant in the mice immuned by pcDNA-CEA625-667 and pcDNA-triCEA625-667 in a shorter time by contrast with na ?ve mice.Low tilter IgG antibody against CEA was detected in the antiserum of the mice immuned by repeats of minigene DNA vaccine , which suggested the activation of helper T-cell.ELISA showed that the level of IFNγin the 3 days culture of the splenocytes was rela-tively higher in the groups of minigene DNA vaccination than in the control groups ,while IL-4 expression was absent in all groups .The immune response level elicited by three tandem repeats of minigene DNA vaccine pcDNA -triCEA625-667 was superior to that elicited by pcDNA-CEA625-667 ,which showed that any immunogenic inadequacies in minigene presentation can be rectified by linking itself in a string-of-beads vaccine.Conclusion:The haploid vaccine and three tandem repeats of minigene DNA vaccine derived from CEA geneboth can not change the ratio of CD 4+/CD8+but can induce the activation of helper T-cell and skew T -cells toward Th -1 response.The immune response level elicited by three tandem repeats of minigene DNA vaccine was superior to thatelicited by haploid vaccine .
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Objective:To research rmIL-18 in vitro culture system CCs induce tumor-specific cytotoxic T lymphocytes CTL and anti-tumor effect in mice.Methods:Used Stem SepTM immune magnetic cells separation method to culture mouse spleen NK cells,T cells and DCs,established culture systems in vitro;used of different approaches,different doses rmIL-18 to immunize HCC tumor-bearing mice,researched the effect of rmIL-18 on tumor growth rate and survival time.Results:rmIL-18 could induce and promote tumor-specific CTL-mediated killing effects in vitro culture system;tumor-specific CTL could significantly inhibit tumor growth(P<0.01) of and prolong the survival time of liver cancer tumor-bearing mice(P<0.01),and the effect was increased with rmIL-18 concentration increased(P<0.01),and intratumoral injection was superior to intraperitoneal injection(P<0.01).Conclusion:rmIL-18 can induce tumor-specific CTL in vitro and play a role in anti-liver cancer in mice.
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Objective:To probe the expression of miR-126 and VEGF in breast cancer, and the anti-tumor effect of miR-126. Methods:The expression of miR-126 and VEGF in breast cancer tissues and cells were detected by qRT-PCR and Western blot;after transfection with miR-126 mimics into MDA-MB-231,expression of VEGF was detected again,MTT assay and cell scratch test were used to verify the influence of miR-126 on proliferation and migration of tumor cells. Results: The expression of miR-126 was lower in the breast cancer tissues and cells,the expression of VEGF was negative correlation with it,increasing the expression of miR-126 may decrease the expression of VEGF and inhibit the proliferation and migration of breast cancer cells. Conclusion:miR-126 can reduce the proliferation and migration of breast cancer cells by inhibiting the expression of VEGF,which play an anti-tumor effect.
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Objective To evaluate the mechanical stability of alternative reconstruction methods after total en bloc spon?dylectomy in the lower lumbar spine. Methods Eight adult fresh cadaveric lumbosacral spines (L1-S1) were adopted. Total en bloc spondylecotmy of the L4 vertebra was performed after intact testing. Four designed reconstruction samples were tested for the range of motion (ROM) of the spine:1) expandable artificial vertebral body and short posterior instrumentation at L3-L5 (SP), 2) ex?pandable artificial vertebral body and short posterior instrumentation with additional anterolateral fixation at L 3-L5 (ASP), 3) ex?pandable artificial vertebral body and multilevel posterior instrumentation at L2-S1 (MP), 4) expandable artificial vertebral body and multilevel posterior instrumentation with additional anterolateral fixation at L2-S1 (AMP). Nondestructive biomechanical test?ing was performed on each construct under loading control. The ROM for each construct was obtained by applying pure moments in flexion, extension, lateral bending, and axial rotation. Results In flexion, extension and lateral bending, the ROM of all the re?constructed constructs significantly decreased compared to the intact. The ROM of specimens with anterolateral fixation was less when compared to the ones without additional fixation. In lateral bending, MP (L:1.87° ± 0.32° , R:1.97° ± 0.33° ), ASP (L:1.89° ± 0.37°, R:2.08°±0.36°) and AMP (L:1.32°±0.29°, R:1.61°±0.33°) provided significantly less ROM than the SP (L:3.14°±0.35°, R:3.44°±0.34°). In axial rotation, the ROM of ASP (L:4.21°±0.58°, R:4.02°±0.59°) and AMP (L:3.56°±0.55°, R:3.52°±0.48°) was significant decreased when compared to the intact state (L: 7.47° ± 1.00° , R:7.57° ± 0.84° ). MP (L:6.33° ± 0.71° , R:5.88° ± 0.62°), ASP and AMP showed significantly less ROM than the short posterior fixation (L:9.28°±1.01°, R:9.48°±0.98°). AMP sig?nificantly decreased the ROM compared to MP. Conclusion After total en bloc spondylectomy of lower lumbar, long segmental fixation provided more stability to the construct than the short one. Compared to posterior fixation, circumferential fixations showed a higher stability. In contrast, multilevel segmental instrumentation with circumferential fixation did not provided more stability than the short constructs.
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Objective To investigate the effect of tannic acid on glomerular mesangial cells (GMC),and to clarify the mechanism of tannic acid in improving the pathological changes of diabetic nephropathy (DN)from the aspect of oxidative stress and micro-inflammation. Methods The glomerular mesangial cells were treated with glucose (30 mmol·L-1 )or advanced glycosylation end-products (AGEs)bovine serum albumin(BSA)(250 mg·L-1 )and then different concentrations of tannic acid (10,20,40 and 80μmol·L-1 )were added into the GMC.The cells cultured by normal glucose or treated with BSA were used as control groups and then the level of malonic dialdehyde (MDA), glutathione peroxidase (GSH-Px ), superoxide Dismutase (SOD ), CAT (Catalase ) activities and 8-hydroxy-2′-deoxyguanosine(8-OHdG)levels in the culture supernatant 48 h after culture were determined by colorimetry and ELISA method. The expressions of intercellular cell adhesion molecule-1 (ICAM-1 ) protein, monocyte chemotactic protein 1 (MCP-1 ) and ICAM-1 mRNA in GMC were detected by immunohistochemical staining and RT-PCR method.Results Compared with high glucose and AGEs groups,the MDA levels in tannic acid groups were reduced significantly(P<0.05);the activities of GSH-Px,SOD and CAT were increased significantly(P<0.05 or P<0.01);the 8-OHdG levels in annic acid groups were significantly reduced (P<0.05). Compared with high glucose and AGEs groups,the expressions levels of ICAM-1 protein in 40 and 80μmol· L-1 tannic acid groups were decreased (P<0.05 ). The mRNA expressions levels of MCP-1 and ICAM-1 were significantly lower than those in high glucose group (P<0.01 ).Conclusion Tannic acid could protect GMC against the damage of oxidative and inflammatory mediators,thereby delaying and improving the glomerular lesions of DN.
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Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt)inhibitor LY294002,and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group. The inhibitory rate of growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis. The expression levels of Ser473 phosphorylated form of Akt (p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method. Results The inhibitory rate of growth of the A549 cells in LY294002+rsTRAIL group (74.6 %± 2.63%)was higher than that in rsTRAIL group (5.61% ± 0.32%) (P< 0.05 ). Compared with rsTRAIL group, the percentage of the cells at G0/G1 phase in LY294002+rsTRAIL group was increased(P<0.05)and the percentage of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%)was higher than that in rsTRAIL group (3.21%±0.96%)(P<0.05). The Western blotting results showed that the expression levels of p-Akt, c-FLIPL and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05 )and the ratio of Bax/Bcl-2 was increased (P<0.05 ) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.
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Objective:To observe the expression of Ki-67,P53 and LAT1 in tissues of esophageal squamous cell carcinoma and precancerous lesions and to investigate its clinical significance.Methods: Immunohistochemical method was used to detect the expressions of Ki-67,P53 and LAT1 in tissues from 20 cases of normal esophageal mucosa,68 cases of precancerous lesions including 21 cases of mild atypical hyperplasia,22 cases of atypical hyperplasia,25 cases of severe atypical hyperplasia and cancer tissues from 34 esophageal cancer patients.The relationship between the expression of the three in esophageal carcinoma was analysed.Results: Re-spectively,the positive expression rate of Ki-67 in normal esophageal mucosa, mild atypical hyperplasia, moderate dysplasia, severe dysplasia and carcinoma was 0%( 0/20 ) , 23.8%( 5/21 ) , 40.9%( 9/22 ) , 76.0%( 19/25 ) , 82.4%( 28/34 ) and the positive expression rate of P53 was 0%(0/20),14.3%(3/21),31.8%(7/22),48.0%(12/25),67.6%(23/34) and the positive expression rate of LAT1 was 0%( 0/20 ) , 19.0%( 4/21 ) , 36.4%( 8/22 ) , 52.0%( 13/25 ) , 76.5%( 26/34 ).The rank correlation analysis showed that the positive expression of Ki-67,P53 and LAT1 were significantly correlated with histological grade(r=0.626,0.427, 0.586,P<0.01) and the expression of Ki-67,P53 was positively correlated with LAT1 in esophageal carcinoma tissues(r=0.428, 0.596,P<0.01).Conclusion:Abnormal expression of Ki-67,P53,LAT1 protein was significant related to carcinogenesis of esophageal cancer and the combined detection of the three has important clinical significance.
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Objective:To observe the effect of tannic acid on renal morphology and function of diabetes mellitus model rats ,and to explore the mechanism of improving effect of tannic acid from oxidative stress , nitrosative stress angle.Methods: 8 rats were randomly selected from 68 6-week-old male Wistar rats as normal control group and the remaining 60 rats accepted high-sugar and high-fat diet for 4 weeks, then were injected streptozotocin ( STZ, 52 mg/kg ) intraperitoneally in order to manufacture a diabetic rat model.Further the diabetic rats were randomly divided into model group ,aminoguanidine group ,low-dose of tannic acid group and high dose of tannic acid group.The rats in aminoguanidine group were injected aminoguanidine [AG,40 mg/(kg· d)] intraperitoneally, those in low-dose of tannic acid group were injected tannic acid [TA,20 mg/(kg· d)] and those in high-dose of tannic acid group were injected tannic acid [TA,30 mg/(kg· d)].The rats in normal control group and model group were injected normal saline [NS, 30 mg/(kg· d)] and all rats were sacrificed and tissues were derived at the end of the week 10.Morphologic changes of kidney in diabetic rats were observed by HE staining and correlative biochemical indices of renal function were detected by biochemical analyzer.8-hydroxy deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) content of renal tissue in rats in different groups were detected by ELISA method.Mesangial cells cultured in vitro were treated with high concentration of glucose (30 mmol/L) and AGEs (250 mg/L) and at the same time with different concentration of tannic acid (10,20,40 and 80μmol/L) on the basis of setting corre-sponding control group.The contents of 8-OHdG and 3-NT in the culture supernatant were measured by ELISA method after 48 hours.Results:Tannic acid can effectively improve the renal pathological changes and improve renal function of diabetic rats .The contents of 8-OHdG and 3-NT in kidney tissue homogenate of diabetic rats and in the supernatant of GMC cultured with high glucose or AGEs were all significantly increased and can be reduced by tannic acid.Conclusion:Tannic acid improving the structure and function damage of kidney in diabetic rats might be achieved by oxidative stress and nitrosative stress mechanism .
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Objective To detected the motor cortex reorganization and compared the influence on reorganization process as regard to different transfer modes of contralateral seventh cervical nerve root (C7)in young rats model of total brachial plexus root avulsion. Methods The young Sprague-Dawley rats model of total brachial plexus root avulsion was established. The left radix dorsalis and radix ventralis from the fifth cervical nerve root (C5) to the first thoracic nerve root (T1)were exposed and the roots from C5 to T1 were avulsed from the spinal cord. Then, the contralateral C7 transfer operation was performed. Three different operative modes were applied randomly which included contralateral C7 transference to anterior division of the upper trunk (group A, n=30), to both musculocutaneous and median nerves (group B, n=30), or to median nerve (group C, n=30). The movement evocation of motor cortex was measured by intracortical microstimula-tion in both hemispheres, and functional reorganization was assessed dynamically in 1.5, 3, 6, 9 and 12 months after operation. Results After contralateral C7 transference, the ipsilateral motor cortex initially acti- vated the injured limb at 1.5 month, and subsequently the motor cortex of both hemispheres activated the in-jured limb at the 3rd and 6th month. The injured limb was activated mainly by the contralateral motor cortex in group A at the 9th month. The contralateral motor cortex exclusively controlled the injured limb in all three groups at the 12th month after the operation. Meanwhile, the extent of functional reorganization was better in group B than that of group C. Conclusion After contralateral C7 transfer operation, motor cortex in charging injured limb developed a transhemispheric functional reorganization in young rats with total brachial plexus root avulsion. Different operative modes contributed to the functional reorganization of motor cortex. Transferring contralateral C7 to anterior division of the upper trunk or to both musculocutaneous and median nerves provided better functional reorganization than to median nerve.
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OBJECTIVE: To study the tongue tissue blood oxygen saturation measurement for evaluating tongue manifestation of blood stasis syndrome, and to explore its correlation with blood rheological disorder in a rat model of acute transient brain ischemia. METHODS: Twenty-eight SD rats were randomly divided into sham-operated group and ischemia group. Middle cerebral artery occlusion was induced by thread in rats of the ischemia group. Tongue tissue blood oxygen saturation, neurological severity score and the changes of blood viscosity, red blood cell deformity, thrombin time and fibrinogen in the rats were measured after 24-hour reperfusion. RESULTS: Blood viscosity and the content of fibrinogen in the ischemia group were significantly higher than those in the sham-operated group. Red blood cell deformity, thrombin time and tongue tissue blood oxygen saturation in the ischemia group were decreased as compared with the sham-operated group. There was a positive correlation between red blood cell deformity and tongue tissue blood oxygen saturation. CONCLUSION: Tongue tissue blood oxygen saturation is a good measurement for evaluating blood stasis in a rat model of focal cerebral ischemia, and this model can be used as a rat model of stroke with blood stasis syndrome.
ABSTRACT
[Objective]To study the relationship between ossification of ligamentum nuchae(LN) and degeneration of ligamentum flavon(LF) of cervical spondylotic myelopathy(CSM).[Method]Fifty-six cervical LF samples from CSM patient of C_(3~7) in relation to the stage of ossification of LN samples(experimental group,EG) and 38 samples of corresponding ligament from patient with cervical vertebral trauma(CVT)(control group,CG) were obtained.Thickness of LF was measured,their pathological change was examined by microscopy,content of collagen and kydrolyproline and the ratio of type Ⅰ/ type Ⅱ collagen were determined by Woessners method and setting Salting out method.The correlation among the thickness of degenerative LF,the ratio of type Ⅰ/ type Ⅱ collagen,the content of collagen and hydrokyproline,and arrangement disorder of LF in relation to the ossification stage of LN were analyzed.[Result]In EG,as compared with in CG,there showed a decrease of elastic fibre,decrease of ratio type Ⅰ/ type Ⅱ collagen due to significant increase of type Ⅱ collagen,increase collagen content and disorder arrangement of LF.In relation to the ossification of LN,above changes were more marked in LF of C_(4、5),C_(5、6),than that of C_(3、4),C_(6、7)(P