[Real-time localization for port-implanted catheter tip by echocardiographic guidance].

The clinical data of 23 patients undergoing real-time echocardiography-guided infusion port implantation in the Breast Center of Tsinghua Changgung Hospital in Beijing from January to July 2021 were analyzed. The length of catheter insertion L1 was initially estimated using surface measurement method in all patients. Intraoperatively, transthoracic echocardiography was applied using the parasternal four-chamber view to visualize the catheter image within the right atrium, and the length of catheter insertion L2 was recorded under the guidance of echocardiography. Postoperatively, chest radiographs were taken in the upright position to observe the position of the catheter tip. According to chest CT scans, the ideal length (L) for catheter tip placement was calculated when it was located at the junction of superior vena cava and right atrium. Bland-Altman scatter plot analysis and linear regression fitting test were used on L1 and L2 respectively with L to evaluate the consistency. A total of 23 patients were included in this study, among which one case of left breast cancer patient undergoing breast-conserving surgery had difficulty in identifying the catheter tip position due to residual pleural effusion obscuring the imaging of the cardiac apex four-chamber view. In 22 patients, the results of intraoperative ultrasound imaging were good, including 1 case of catheter ectopic to azygos vein, and 21 cases of right atrial catheter could be detected by ultrasound. Statistical analysis showed that there was a good consistency between L1 and L, L2 and L, and the difference between them was d=0.28 cm (95%CI:-1.76-2.31 cm) and d=0.20 cm(95%CI:-0.84-1.23 cm), respectively, with no statistical significance (P>0.05). In the linear regression model, L2 and L had a higher fit than L1, and the difference was statistically significant (R²=0.954, P<0.001). This study found that real-time echocardiographic localization technique can be applied in adult port surgery to replace X-ray-guided real-time catheter tip detection and adjustment to the optimal position.

[Application of transbronchial cryobiopsy in the diagnosis of postoperative complications after lung transplantation: a report of 6 cases].

Objective: To investigate the efficacy and safety of transbronchial cryobiopsy (TBCB) after lung transplantation. Methods: The clinical characteristics, TBCB procedure, diagnosis and treatment, and outcomes of lung transplant recipients of 6 patients (all male, aged 33-67 years) with TBCB in China-Japan Friendship Hospital from May to November 2021 were retrospectively analyzed. Results: Among the 6 patients diagnosed by TBCB, there were 2 cases of organizing pneumonia, 1 acute cellular rejection, 1 antibody-mediated rejection, and 1 bronchiolitis obliterans, and 1 diffuse alveolar damage. After the clinical diagnosis was confirmed, the condition improved after adjustment of the treatments followed. There were no serious complications related to the TBCB procedure. Conclusion: TBCB is valuable and relatively safe in the diagnosis of complications after lung transplantation, but the indications need to be strictly controlled.

[Pneumatosis cystoides intestinalis in lung transplant recipients: three cases report and literature review].

Objective: To report the clinical characteristics and treatment courses of pneumatosis cystoides intestinalis(PCI) after lung transplantation(LT). Methods: We included all cases of PCI after LT from March 2017 to June 2021 in China-Japan Friendship Hospital. In addition to our cases, we searched literatures published in Chinese and English languages using China National Knowledge Infrastructure (CNKI), Wanfang Data and PubMed/MEDLINE with the search terms"pneumatosis intestinalis"and"lung transplantation". The clinical characteristics and treatment courses of all cases were summarized and analyzed. Results: Three cases of PCI occurred after LT in this study, with an incidence of 0.804% (3/373). Thirteen related literatures were retrieved, with 51 cases enrolled. The median age of the 54 patients was 55.4 years (22-79 years), with 33 males and 21 females. 64.81% (35/54) of the 54 patients underwent LT for interstitial lung disease and 90.74% (49/54) underwent bilateral LT. Twenty-two cases(40.7%) were asymptomatic when PCI occurred. Thirty-eight cases (38/54,70.37%)had involvement of ascending colon, and 35 cases(35/54,64.81%)had involvement of transverse colon. Forty-three cases(43/54, 79.63%) were treated conservatively. The average interval between transplantation and PCI was 210 (5-2 495) days. Conclusion: PCI is a rare complication after lung transplantation, most often occurring in the colon. Most patients were asymptomatic and could improve by conservative treatments.

[Clinical value of TDI combined with 2D-STI on evaluating the microcirculation dysfunction and left ventricular dysfunction in patients with non-obstructive coronary angina].

Objective: To explore the value of tissue Doppler imaging (TDI) combined with two-dimensional speckle tracking imaging (2D-STI) at rest on evaluating microcirculation dysfunction and left ventricular dysfunction in patients with angina and no obstructive coronary artery disease(ANOCA). Methods: This retrospective study recruited 78 ANOCA patients, who hospitalized in the People's Hospital of Liaoning Province from August 2019 to July 2021. These patients underwent conventional echocardiography examination, including TDI and 2D-STI, to evaluate the left ventricular dysfunction, and adenosine stress echocardiography (SE) to evaluate the coronary flow velocity reserve (CFVR). ANOCA patients were divided into coronary microcirculation dysfunction CMD group (CFVR<2) and control group (CFVR≥2) according to CFVR. Clinical data, routine echocardiographic parameters, TDI parameters including isovolumic contraction time (IVCT), isovolumic relaxation time (IVRT), ejection time (ET), and STI parameters including global longitudinal peak strain (GLS), time to peak (TTP); peak strain dispersion (PSD) were compared between the two groups. Binary logistic regression was used to analyze the risk factors of CMD and the predictive value of each parameter to construct a joint prediction model for the diagnosis of CMD in this patient cohort. Results: The mean age was (55.5±11.2) years, 43 (55%) patients were females in this patient cohort, 38 (49%) patienst were didvided into the CMD group and 40 (51%) into the control group. Age, prevalence of hypertension, diabetes, dyslipidemia, and smokers were significantly higher in the CMD group than in the control group (all P<0.05). Tei index was higher, IVCT and TTP were longer, PSD was higher, ET was shorter, and absolute GLS was lower in the CMD group than in the control group (all P<0.05). The results of logistic regression analysis showed that longer IVCT, higher Tei index, higher time to PSD and lower absolute GLS were the independent risk factors of CMD. The ROC curve revealed that the predicting efficacy on CMD was satisfactiory with the combined predictors: AUC=0.884, sensitivity of 82% and specificity of 80%. Conclusions: TDI combined with 2D-STI is associated with a good diagnostic value on the diagnosis of CMD and left ventricular dysfunction in patients with ANOCA, which provides a feasible non-invasive tool for the diagnosis of CMD and risk stratification of patients with ANOCA.

[Pulmonary mucormycosis after lung transplantation:3 cases report with literature review].

Objective: To report the risk factors, clinical characteristics and treatment courses of pulmonary mucormycosis after lung transplantation(LT). Methods: We included 3 cases with pulmonary mucormycosis after LT from March 2017 to July 2020 in the centre for lung transplantation of China-Japan Friendship Hospital. Twelve cases from Chinese and English literature from China National Knowledge Infrastructure (CNKI), China Biomedical Literature Service System and Pubmed Database from March 1980 to July 2020 were added. The risk factors, clinical characteristics and treatment courses of all cases were summarized and analyzed. Results: Pulmonary mucormycosis occurred in 1.06% (3/284) in our centre. A total of 15 cases with 12 cases from literature included 10 males and 5 females with a mean age of(47±20)years. Thirteen cases occurred after LT, and 2 cases occurred after heart-lung transplantation (HLT). Nine probable cases were diagnosed by positive isolation of the pathogen from bronchoalveolar lavage fluid or sputum. Three proven cases were diagnosed by transbronchial lung biopsy. Meanwhile, the other 3 proven cases diagnosed by CT-guided percutaneous lung biopsy, autopsy and surgical operation respectively. Ten cases (66.7%) were diagnosed with pulmonary mucormycosis within 90 days after lung transplantation. The mortality was as high as 46.67% (7/15), but if it occurred within 90 days, the mortality reached 70% (7/10). The average interval between transplantation and positive isolation of the pathogen was 112.3 (5-378) days. Conclusions: The clinical and radiographic features of pulmonary mucormycosis after LT were nonspecific. It had a high mortality, especially in those occurred within 90 days after LT. The combination of antifungal therapy and surgical resection may contribute to a better outcome of the disease.

[Prevalence of CYP2C19 gene mutations in patients with coronary heart disease and its biological activation effect in clopidogrel antiplatelet response].

Objective: The purpose of this study was to investigate the effects of CYP2C19 gene mutations on clopidogrel antiplatelet activity in the patients with coronary heart disease treated by percutaneous coronary intervention. Methods: Patients with coronary heart disease, who hospitalized in the Second Affiliated Hospital of Nanchang University from March 2011 to June 2019, and healthy individuals with matching genetic background, gender, and age as controls were included in this study. Basic clinical data were analyzed and blood samples of all research subjects were obtained for extraction of DNA, and Sanger first-generation sequencing method was used to detect CYP2C19 gene mutation from full exon and exon and intron junction. CYP2C19 gene variations in patients with coronary heart disease were compared with the 1000 Genomes Browse database and the sequencing results of healthy controls to determine whether the gene variation was a genetic mutation or a genetic polymorphism. After that, PolyPhen-2 prediction software was used to analyze the harmfulness of gene mutations to predict the effect of mutations on protein function. The same dose of CYP2C19 wild-type plasmid and the CYP2C19 gene mutant plasmids were transfected into human normal liver cells HL-7702. After transfection of 24 h, the expression of CYP2C19 protease in each group was detected. The liver S9 protein was incubated with clopidogrel, acted on platelets to detect the platelet aggregation rate and the activity of human vasodilator-activated phosphoprotein (VASP). Results: A total of 1 493 patients with coronary heart disease (59.36%) were enrolled, the average age was (64.5±10.4) years old, of which 1 129 were male (75.62%). Meanwhile, 1 022 healthy physical examination volunteers (40.64%) were enrolled, and the average age was (64.1±11.0) years old, of which 778 were male (76.13%). A total of 5 gene mutations of CYP2C19 gene were identified in 12 patients (0.80%), namely, 4 known mutations T130K (1 case), M136K (6 cases), N277K (3 cases), V472I (1 case) and one new mutation G27V (1 case), no corresponding gene mutation was found in healthy controls. It was found that T130K and M136K were probably damaging, G27V was possibly damaging, and N277K and V472I were benign mutations. In vitro, we demonstrated that the platelet aggregation rate of the M136K gene mutation group was 24.83% lower than that of the wild type (59.58% vs. 34.75%; P<0.05), and the phosphorylated VASP level was 23.0% higher than that of the wild type (1.0 vs. 1.23; P<0.05). However, the platelet aggregation rate and phosphorylated VASP level were similar between of G27V, T130K, N277K, V472I gene mutation groups and wild type group (P>0.05). Conclusions: In this study, 5 gene mutations are defined in patients with coronary heart disease, namely G27V, T130K, M136K, N277K, V472I. In vitro functional studies show that CYP2C19 gene mutation M136K, as a gain-of-function gene mutation, can enhance the activation of CYP2C19 enzyme on clopidogrel, thereby inhibiting the platelet aggregation rate.

[Hypoxia and inflammation are risk factors for acute myocardial injury in patients with coronavirus disease 2019].

OBJECTIVE: To investigate the risk factors for acute myocardial injury in coronavirus disease 2019 (COVID-19) patients. METHODS: This is a retrospective analysis of a COVID-19 cohort, in which 149 confirmed COVID-19 patients enrolled were divided into the group of myocardial injury (19 cases) and the group of non-myocardial injury (130 cases). Myocardial injury was defined according to Fourth universal definition of myocardial infarction released by European Society of Cardiology (ESC) in 2018, that cardiac troponin (cTn) was above 99th percentile of the reference level. Clinical information and results of laboratory tests of the eligible patients were collected. Factors associated with myocardial injury in COVID-19 patients were evaluated. RESULTS: Compared with the group of non-injury, the patients in the group of injury were older and had a larger proportion of severe or critical cases (P < 0.05), higher respiratory rate and lower percutaneous oxygen saturation (SpO2) without oxygen therapy on admission (P < 0.05). All inflammatory indexes except for tumor necrosis factor α (TNF-α) showed significant elevation in the patients of the group of injury (P < 0.05). Analyzed by Spearman correlation test, we showed that the levels of circulatory cTnI were in positive correlation with the levels of high-sensitivity C-reactive protein (hs-CRP), ferritin, receptor of interleukin-2 (IL-2R), interleukin-6 (IL-6) and interleukin-8 (IL-8) (ρ > 0, P < 0.05). Lower SpO2 without oxygen therapy on admission (OR: 0.860, 95%CI: 0.779-0.949, P=0.003) and higher plasma IL-6 levels (OR: 1.068, 95%CI: 1.019-1.120, P=0.006) were independent risk factors for acute myocardial injury in the patients with COVID-19 by multivariate Logistic regression analyses. CONCLUSION: Hypoxic state and inflammation may play a key role in the pathogenesis of acute myocardial injury in COVID-19 patients.

MicroRNA-1271 inhibits the progression of papillary thyroid carcinoma by targeting IRS1 and inactivating AKT pathway.

OBJECTIVE: The important role of microRNA-1271 (miR-1271) has been identified in human diseases and cancers. However, the biological function of miR-1271 remains ambiguous in papillary thyroid carcinoma (PTC). Therefore, the specific role of miR-1271 was investigated in PTC. PATIENTS AND METHODS: The expressions of miR-1271 and insulin receptor substrate 1 (IRS1) were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. The protein expression of the genes was measured by Western blot analysis. The function of miR-1271 was investigated using methyl thiazolyl tetrazolium (MTT) and transwell assays. The Dual-Luciferase assay was used to observe the relationship between miR-1271 and IRS1. RESULTS: MiR-1271 was downregulated in PTC tissues. Moreover, overexpression of miR-1271 suppressed migration, invasion and proliferation of PTC cells. Furthermore, IRS1 was indicated as a direct target gene of miR-1271 and knockdown of IRS1 inhibited cell migration, invasion and proliferation in PTC. In addition, miR-1271 inhibited the progression of PTC by targeting IRS1. Besides that, miR-1271 blocked the epithelial-mesenchymal transition (EMT) and protein kinase B (AKT) pathway in PTC. CONCLUSIONS: MiR-1271 inhibited the progression of PTC by targeting IRS1 and blocking EMT and AKT pathway.

Long non-coding RNA ENST00000434223 inhibits the progression of renal cancer through Wnt/hygro-catenin signaling pathway.

OBJECTIVE: The purpose of this study was to determine the function of long non-coding RNA (LncRNA) ENST00000434223 (Lnc ENST) in renal carcinoma, and to explore the potential molecular mechanism. PATIENTS AND METHODS: Quantitative Real-Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expressions of lncRNA ENST00000434223 and Wnt/ß-catenin pathway-related mRNAs in tissues and cells of renal cancer. Chi-square test was performed to figure out the relationship between lncRNA ENST00000434223 and clinic-pathologic features of renal cancer patients. Besides, si-NC, si-ENST00000434223, pcDNA-NC and pcDNA-ENST00000434223 were transfected into renal cancer cells. The proliferative ability, metastasis and invasiveness of cells were detected using Cell Counting Kit-8 (CCK-8) and transwell assay, respectively. Lastly, the activation of the Wnt/hygro-catenin signal transduction pathway was evaluated by TOP/FOP Wnt Luciferase reporter assay and Western blot. RESULTS: The expressions of Wnt2b and ß-catenin were significantly increased in renal carcinoma, while E-cadherin was markedly down-regulated. Lowly expressed ENST00000434223 was involved in the poor prognosis of patients with renal cancer. In addition, down-regulating ENST00000434223 could enhance the viability, metastasis and invasiveness of renal cancer cells. However, overexpressing ENST00000434223 remarkably weakened the above cell functions. At the same time, interference or overexpression of ENST00000434223 could affect the expression level of proteins related to the Wnt/ß-catenin signal pathway. CONCLUSIONS: LncRNA ENST00000434223 inhibits the progression of renal cancer through the Wnt/shell-catenin signal pathway.

[The application of cortical and subcortical stimulation threshold in identifying the motor pathway and guiding the resection of gliomas in the functional areas].

Objective: This study aimed to analyze the application of cortical and subcortical stimulation threshold in identifying the motor pathway and guiding the resection of gliomas in the functional area, and to illustrate the minimal safe threshold by ROC method. Methods: Fifty-seven patients with gliomas in the functional areas were enrolled in the study at Beijing Tiantan Hospital from 2015 to 2017. Anesthesia was maintained intravenously with propofol 10% and remifentanil. Throughout the resection process, cortical or subcortical stimulation threshold was determined along tumor border using monopolar or bipolar electrodes. The motor pathway was identified and protected from resection according to the stimulation threshold and transcranial MEPs. Minimal threshold in each case was recorded. Results: Total resection was achieved in 32 cases(56.1%), sub-total resection in 22 cases(38.6%), and partial resection in 3 cases(5.3%). Pre-operative motor disability was found in 9 cases. Compared with pre-operative motor scores, 19 exhibited impaired motor functions on day 1 after surgery, 5 had quick recovery by day 7 after surgery, and 7 had late recovery by 3 months after surgery. At 3 months, 7 still had impaired motor function. The frequency of intraoperative seizure was 1.8%(1/57). No other side effect was found during electronic monitoring in the operation. The ROC curve revealed that the minimal safe monopolar subcortical threshold was 5.70 mA for strength deterioration on day 1 and day 7 after surgery. Univariate analysis revealed that decreased transcranial MEPs and minimal subcortical threshold ≤5.7 mA were correlated with postoperative strength deterioration. Conclusions: Cortical and subcortical stimulation threshold has its merit in identifying the motor pathway and guiding the resection for tumors within the functional areas. 5.7 mA can be used as the minimal safe threshold to protect the motor pathway from injury.

[Feasibility and value of index of microcirculatory resistance in patients with acute myocardial infarction after primary percutaneous coronary intervention].

Objective: To evaluate the feasibility of detecting index of microcirculatory resistance (IMR) and the relationship between IMR and left ventricular (LV) systolic function after acute myocardial infarction (AMI) undergoing primary percutaneous coronary intervention (PCI). Methods: The patients with first AMI received primary PCI in Peking University Third Hospital were enrolled from January 2014 to March 2016. IMR were measured immediately after PCI by using pressure/temperature wire. The relationship between IMR and left ventricular ejection fraction (LVEF) assessed by echocardiography at first day and 6 months after admission was evaluated. Results: Twenty-eight patients with anterior wall AMI were enrolled, with an average age (56±13) years. The success rate of IMR detection was 100%. The mean IMR was (33±18 )mmHg·s. There was no complication related to intravenous adenosine triphosphate (ATP) (140 µg· kg(-1)· min(-1)). The IMR was negatively correlated with TIMI blood flow grade after primary PCI (r=-0.386, P=0.043), and positively correlated with female gender, CK peak value and TnT peak value (r=0.430, P=0.022; r=0.431, P=0.025; r=0.434, P=0.024). After 6 months of follow-up, no adverse cardiovascular events (including cardiac death, nonfatal myocardial infarction, malignant arrhythmia, unplanned revascularization, hospitalization for unstable angina pectoris and severe heart failure requiring hospitalization) occurred. LVEF increased significantly compared with the first day after PCI (0.54±0.08 vs 0.47±0.06, P=0.001), and IMR was negatively correlated with LVEF after 6 months (r=-0.477, P=0.014). Multivariable linear regression analysis showed that CK peak and IMR were predictors of LVEF after six months ( ß=-0.595, t=-3.814, P=0.01; ß=-0.352, t=-2.26, P=0.036). Conclusions: Immediate detection of IMR in patients with anterior wall AMI after PCI is safe and feasible. The immediate IMR after PCI reflects the extent of myocardial necrosis and myocardial perfusion, and is a predictor of LVEF at 6 months after PCI.

[Myocardial protection of remote ischemic postconditioning during primary percutaneous coronary intervention in patients with acute ST-segment elevation myocardial infarction].

OBJECTIVE: To evaluate the cardioprotection of remote ischemic postconditioning (RIPostC) in patients with acute ST-segment elevation myocardial infarction (STEMI) who underwent primary percutaneous coronary intervention (PCI). METHODS: Forty-six STEMI patients undergoing primary PCI at Peking University Third Hospital from January to April 2014 were randomized to RIPostC group (n=23) and control group (n=23).The RIPostC protocol was started within 1 min after reflow by thrombus aspiration or balloon inflation and consisted of 3 cycles of 5 min/5 min ischemia/reperfusion by cuff inflation/deflation of the lower left limb. The enzymatic infarct size, rate of complete ST segment resolution, corrected thrombolysis in myocardial infarction (TIMI) frame count (CTFC) in infarct-related artery (IRA) and plasma levels of malondialdehyde(MDA), endothelin-1(ET-1), tumor necrosis factor α (TNFα) of the two groups were compared. RESULTS: There was no significant difference in enzymatic infarct size between the two groups (P>0.05). The rate of complete ST-segment resolution was significantly higher in RIPostC group than in control group (60.9%vs. 30.4%,P=0.04). There was a trend toward lower CTFC in RIPostC group than that in control group, but the difference was not statistically significant(28 ± 11 vs. 33 ± 11, P = 0.10). However, in the subgroup of anterior wall myocardial infarction CTFC in RIPostC group was significantly lower, compared with control group (25±9 vs. 39±10, P=0.01).There were lower plasma levels of MDA,ET-1,TNFα in RIPostC group than in control group at different time points after primary PCI (P<0.05). CONCLUSION: In STEMI patients undergoing primary PCI, RIPostC may improve myocardial perfusion and attenuate ischemia reperfusion injury with the underlying mechanisms involving reduction of oxidative stress, protection of endothelial function and inhibition of inflammatory response.

Long-term inhibition of Rho-kinase restores the LTP impaired in chronic forebrain ischemia rats by regulating GABAA and GABAB receptors.

We previously demonstrated that inactivation of Rho-kinase by hydroxyfasudil could impact N-methyl-d-aspartate (NMDA) excitatory interneurons in the hippocampus and attenuate the spatial learning and memory dysfunction of rats caused by chronic forebrain hypoperfusion ischemia. Complementary interactions between the excitatory neurotransmitter glutamate and the inhibitory neurotransmitter GABA form the molecular basis of synaptic plasticity and cognitive performance. However, whether the GABAergic inhibitory interneurons are involved in the mechanisms underlying these processes remains unclear. Here, we further examined the role of GABAergic interneurons in the neuroprotective effect of the Rho-kinase inhibitor. Chronic forebrain ischemia was induced in Wistar rats by bilateral common carotid artery occlusion (BCAO). The general synaptic transmission and long-term potentiation (LTP) of hippocampal CA3 neurons were evaluated at 30 days after sham surgery or BCAO. Real-time PCR and Western blot analyses were conducted to determine the effect of the Rho-kinase inhibitor hydroxyfasudil on GABAergic inhibitory interneuron expression and function after ischemia. Hydroxyfasudil showed no significant effect on general synaptic transmission, but it could abolish the inhibition of LTP induced by chronic forebrain ischemia. Moreover, the mRNA and protein levels of GABAA and GABAB in three brain regions after ischemia were markedly decreased, and hydroxyfasudil could up-regulate all mRNA and protein expression levels in these areas except for GABAA mRNA in the cerebral cortex and striatum. Using phosphorylation antibodies against specific sites on the GABAA and GABAB receptors, we further demonstrated that hydroxyfasudil could inhibit GABAergic interneuron phosphorylation triggered by the theta burst stimulation. In summary, our results indicated that the inactivation of Rho-kinase could enhance GABAA and GABAB expressions by different mechanisms to guarantee the induction of hippocampal LTP, and it could decrease the phosphorylation level of GABAergic inhibitory interneurons to promote the LTP induction rate and magnitude, hence improving the cognitive deficit suffered after chronic forebrain ischemia.

A study on dysbaric osteonecrosis in caisson workers.

PURPOSE: To study the effects of exposure to compressed air on tunnel workers' health and to investigate the prevalence of dysbaric osteonecrosis (DON) in caisson workers. METHODS: 128 tunnel workers were divided into the exposed group (n = 58) and the control group (n = 70), and their shoulders, hips and knees were examined with X-ray, computed tomography (CT) and magnetic resonance imaging (MRI). RESULTS: 1) 34.5% of the exposed group were diagnosed with DON based on the national diagnostic criteria of decompression sickness. 2) The incidental difference of skeletal cystic changes between the exposed group and the control group was highly statistically significant (p < 0.01). 3) CT and MRI examination could detect early onset of DON lesions, and the cystic changes shown in CT and abnormal signals in MRI were diagnostic indicators in cases. CONCLUSION: Cystic changes in CT and abnormal signals in MRI are key imaging findings of early DON.

The alteration of Id-1 and TSP-1 expression in mucoepidermoid carcinoma associated with its clinical features and prognosis.

Expression of Id-1 (inhibitors of DNA binding/differentiation protein 1) and TSP-1 (thrombospondin-1) in mucoepidermoid carcinoma and their relationship to pathological features and prognosis was studied. Moderately and poorly differentiated groups had significantly higher Id-1 positive expression rate (p<0.05) than well differentiated carcinoma. Stages III-IV showed significant increase of Id-1 positive expression rate (p<0.05) compared with stages I and II. Id-1 positive expression was significantly higher in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After that, patients with negative Id-1 expression had significantly higher tumor-free survival than patients with positive expression (p<0.05). Correlation between the expression of Id-1 and TSP-1 in mucoepidermoid carcinoma was negative (p<0.05). Poorly differentiated groups show significantly lower TSP-1 positive expression rate than well differentiated groups (p<0.05). No significant differences of TSP-1 positive expression were detected with clinical stage. TSP-1 positive expression was significantly lower in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After 5 years, patients with positive TSP-1 expression had significantly higher tumor-free survival than patients with negative TSP-1. Positive Id-1 expression is associated with high malignancy/poor prognosis; positive TSP-1 expression is associated with low malignancy/good prognosis. Protein expression status may help assess tumor malignancy and patient prognosis.

Recombinant tissue metalloproteinase inhibitor-3 protein induces apoptosis of murine osteoblast MC3T3-E1.

Tissue inhibitor of metalloproteinases (TIMPs) plays an essential role in the regulation of bone metabolism. Here we report that recombinant tissue metalloproteinase inhibitor-3 (TIMP-3) protein induces the apoptosis of MC3T3-E1 osteoblasts. Cell apoptosis was detected by sandwich-enzyme-immunoassay. Fas and Fasl protein levels were determined by Western blot analysis. The enzyme substrate was used to assess the activation of caspase-3 and caspase-8. The phosphorylation of JNK, p38 and ERK1/2 was examined by Western blot analysis. The ELISA suggested that TIMP-3 promoted MC3T3-E1 cell apoptosis. TIMP-3 treatment induced the expression of Fas and Fasl proteins, and the activation of caspase-8 and caspase-3. TIMP-3 treatment induced p38 and ERK phosphorylation. SB203580 and PD98059, the inhibitor of p38 and ERK, respectively, abolished the TIMP-3 effect on osteoblast apoptosis. In conclusion, the signal pathway through which TIMP-3 induces MC3T3-E1 cell apoptosis, mediated by Fas and involves the p38 and ERK signal transduction pathways.

Tissue inhibitor of matrix metalloproteinase-1 suppresses apoptosis of mouse bone marrow stromal cell line MBA-1.

We investigated the action of tissue inhibitor of metalloproteinase-1 (TIMP-1) on apoptosis and differentiation of mouse bone marrow stromal cell line MBA-1. TIMP-1 did not affect alkaline phosphatase (ALP) activity, suggesting that it is not involved in osteoblastic differentiation in MBA-1 cells. However, TIMP-1 inhibited MBA-1 apoptosis induced by serum deprivation in a dose-dependent manner. Our study also showed increased Bcl-2 protein expression and decreased Bax protein expression with TIMP-1 treatment. TIMP-1 decreased cytochrome c release and caspase-3 activation in MBA-1 cells. TIMP-1 activated phosphatidylinositol 3-kinase (PI3-kinase) and c-Jun N-terminal kinase (JNK), and the PI3-kinase inhibitor LY294002 or the JNK inhibitor SP600125 abolished its antiapoptotic activity. To investigate whether antiapoptotic action of TIMP-1 was mediated through its inhibition on MMP activities, we constructed mutant TIMP-1 by side-directed mutagenesis, which abolished the inhibitory activity of MMPs by deletion of Cys1 to Ala4. Wild-type TIMP-1 and mutant TIMP-1 expression plasmids were transfected in MBA-1 cells, and results showed that mutant TIMP-1 still protected the induced MBA-1 cell against apoptosis. These data suggest that TIMP-1 antiapoptotic actions are mediated via the PI3-kinase and JNK signaling pathways and independent of TIMP-1 inhibition of MMP activities.

Membrane-type matrix metalloproteinase-1 (MT1-MMP) is down-regulated in estrogen-deficient rat osteoblast in vivo.

Our previous study showed that estrogen stimulates membrane-type matrix metalloproteinases-1 (MT1-MMP) production in osteoblastic cells culture, but has no effect on MMP-2 and TIMP-2 synthesis. Osteoblast-derived MT1-MMP have been recently implied to play a role in bone metabolism by degrading tumor necrosis factor-alpha (TNF-alpha), resolving extracellular matrix and activating proMMP-2, which requires the process of activation mediated by MT1-MMP/tissue inhibitor of metalloproteinase (TIMP-2) complex on the cell surface. To investigate the mechanism of bone loss following estrogen deficiency, we examined the effects of estrogen on osteoblast synthesis of MT1-MMP, MMP-2 and TIMP-2. In situ hybridization and immunohistochemistry of rat bone samples were used to document the synthesis of MT1-MMP, MMP-2, and TIMP-2 mRNA and protein. Osteoblasts from distal femoral head showed an increase in the pattern of MT1-MMP mRNA and protein production in sham-operated controls and 17beta-estradiol (E2)-treated rats, compared with the ovariectomized group; the synthesis of MMP-2 and TIMP-2 mRNA and protein was unaffected. Our data show a down-regulation of MT1-MMP synthesis by osteoblast in vivo following estrogen withdrawal, and treatment with E2 resulted in induced MT1-MMP expression in vivo. There is evidence suggesting a role for MT1-MMP in the process of bone loss during the pathogenesis of osteoporosis.