A nomogram model for predicting intramyocardial hemorrhage post-PCI based on SYNTAX score and clinical features.

OBJECTIVE: The aim of this study is to develop a nomogram model for predicting the occurrence of intramyocardial hemorrhage (IMH) in patients with Acute Myocardial Infarction (AMI) following Percutaneous Coronary Intervention (PCI). The model is constructed utilizing clinical data and the SYNTAX Score (SS), and its predictive value is thoroughly evaluated. METHODS: A retrospective study was conducted, including 216 patients with AMI who underwent Cardiac Magnetic Resonance (CMR) within a week post-PCI. Clinical data were collected for all patients, and their SS were calculated based on coronary angiography results. Based on the presence or absence of IMH as indicated by CMR, patients were categorized into two groups: the IMH group (109 patients) and the non-IMH group (107 patients). The patients were randomly divided in a 7:3 ratio into a training set (151 patients) and a validation set (65 patients). A nomogram model was constructed using univariate and multivariate logistic regression analyses. The predictive capability of the model was assessed using Receiver Operating Characteristic (ROC) curve analysis, comparing the predictive value based on the area under the ROC curve (AUC). RESULTS: In the training set, IMH post-PCI was observed in 78 AMI patients on CMR, while 73 did not show IMH. Variables with a significance level of P < 0.05 were screened using univariate logistic regression analysis. Twelve indicators were selected for multivariate logistic regression analysis: heart rate, diastolic blood pressure, ST segment elevation on electrocardiogram, culprit vessel, symptom onset to reperfusion time, C-reactive protein, aspartate aminotransferase, lactate dehydrogenase, creatine kinase, creatine kinase-MB, high-sensitivity troponin T (HS-TnT), and SYNTAX Score. Based on multivariate logistic regression results, two independent predictive factors were identified: HS-TnT (Odds Ratio [OR] = 1.61, 95% Confidence Interval [CI]: 1.21-2.25, P = 0.003) and SS (OR = 2.54, 95% CI: 1.42-4.90, P = 0.003). Consequently, a nomogram model was constructed based on these findings. The AUC of the nomogram model in the training set was 0.893 (95% CI: 0.840-0.946), and in the validation set, it was 0.910 (95% CI: 0.823-0.970). Good consistency and accuracy of the model were demonstrated by calibration and decision curve analysis. CONCLUSION: The nomogram model, constructed utilizing HS-TnT and SS, demonstrates accurate predictive capability for the risk of IMH post-PCI in patients with AMI. This model offers significant guidance and theoretical support for the clinical diagnosis and treatment of these patients.

Serum exosomal hsa_circRNA_0001842 is the potential biomarker for diagnosing lower limb vascular disease in type 2 diabetes mellitus.

This study aimed to identify the potential circular RNAs (circRNAs) in exosomes isolated from serum as biomarkers of lower limb vascular disease (LLVD) in patients with type 2 diabetes mellitus (T2DM). This research collected circRNAs from exosomes isolated from three T2DM patients and three T2DM patients with LLVD for microarray analysis. Five candidate biomarkers derived from differentially expressed circRNAs were then validated by quantitative real-time polymerase chain reaction in 20 T2DM patients and 20 T2DM patients with LLVD. Finally, expression levels of circRNAs were validated in 160 samples. Significant differences in the expression of 295 circRNAs were found between T2DM controls and T2DM patients with LLVD. Among them, 191 differentially expressed circRNAs were upregulated, and 104 were downregulated in T2DM patients with LLVD. Three upregulated and two downregulated circRNAs were further confirmed in 40 samples. According to the testing of 160 samples, hsa_circ_0001842 showed a noticeable specificity in the T2DM patients with LLVD group (n = 80), with an area under the curve (AUC) of 0.79, a sensitivity of 88.75%, and a specificity of 68.75%. In conclusion, hsa_circ_0001842 was found as a potential diagnostic biomarker for T2DM with LLVD.

Efficacy of relative dose intensity of nab-paclitaxel for the short-term outcomes, survival, and quality of life in patients with advanced pancreatic cancer: a retrospective study.

Background: To optimize treatment, choosing the appropriate relative dose intensity (RDI) of nab-paclitaxel is an important way to improve patient tolerance, therapeutic efficacy, and survival. However, few studies have focused on the efficacy of the RDI of nab-paclitaxel in patients with advanced pancreatic cancer, and whether the RDI of nab-paclitaxel could be employed as an index for treatment remains unknown. To explore the relationship between RDI of nab-paclitaxel and chemotherapy efficacy, survival, quality of life (QoL), and adverse effects in patients with advanced pancreatic cancer. Methods: In this retrospective study, a total of 32 patients with advanced pancreatic cancer, ECOG score of 0 to 2 were included from January 2017 to March 2020. The patients were treated with nab-paclitaxel combined with gemcitabine as a first-line treatment and divided into high and low RDI groups. Chemotherapy efficacy, survival, QoL, and adverse effects between two groups were compared. Results: The disease control rate (DCR) was 20.0% in the low RDI group, compared with 81.8% in the high RDI group (P=0.002). A good correlation between nab-paclitaxel RDI and short-term efficacy was observed in all 32 patients (r=0.728, P<0.01). Furthermore, the high RDI group had significantly better median overall survival (mOS: 12 vs. 8 months, P=0.034) and median progression-free survival (mPFS: 5.5 vs. 3 months, P=0.052) compared to that of low RDI patients. Univariate regression analysis showed that longer overall survival was associated with lower ECOG score [hazard ratio (HR): 10.88; 95% confidence interval (CI): 2.54-46.5, P=0.001], tumors located in the body or tail of pancreases (HR: 3.82; 95% CI: 1.4-10.3, P=0.0081), and higher RDI (HR: 0.21; 95% CI: 0.071-0.6, P=0.004). The high RDI group had a significantly better physical function and emotional function improvement compared to the low RDI group (P<0.05). Moreover, high RDI did not increasing the severity and frequency of the adverse events. Conclusions: It is recommended to maintain a sufficient RDI of nab-paclitaxel to ensure that the balance between lerability, therapeutic efficacy, and survival benefits is satisfied in patients with advanced pancreatic cancer.

Development and verification of a hypoxia- and immune-associated prognosis signature for esophageal squamous cell carcinoma.

Background: Esophageal cancer is one of the most common gastrointestinal malignancies worldwide, with high morbidity and mortality in China. The clinical importance of the interaction between hypoxia and immune status in the tumor microenvironment has been established in esophageal squamous cell carcinoma (ESCC). This study aims to develop a new hypoxia- and immune-based gene signature to predict the survival of ESCC patients. Methods: The RNA-sequencing and clinical data of 173 cases of ESCC and 271 normal tissues were obtained from The Cancer Genome Atlas (TCGA) data portal and the Genotype-Tissue Expression (GTEx) database. Hypoxia-related genes (HRGs) and immune-related genes (IRGs) were retrieved from publicly shared data. Differentially expressed gene (DEG) analyses were carried out by the DESeq2 method using the edgeR package in R. Based on the intersection of the DEGs and HRGs/IRGs, differentially expressed HRGs (DEHRGs) and differentially expressed IRGs (DEIRGs) were obtained. DEHRGs and DEIRGs associated with prognosis were evaluated using univariate Cox proportional hazards analysis. A prognostic risk score model was constructed according to the genes acquired through Cox regression. Univariate analysis and Cox proportional hazards analysis were used to determine the independent prognostic factors related to prognosis. A nomogram was developed to predict the 1-, 2-, and 3-year overall survival (OS) probability. Results: A total of 73 intersecting genes were obtained as DEHRGs and a total of 548 intersecting genes were obtained as DEIRGs. The risk score was established using 8 genes (FABP7, TLR1, SYTL1, APLN, OSM, EGFR, IL17RD, MYH9) acquired from univariate Cox analysis. Based on this 8-gene-based risk score, a risk prognosis classifier was constructed to classify the samples into high- and low-risk groups according to the median risk score. The nomogram model was constructed to predict the OS of ESCC patients. Conclusions: The hypoxia- and immune-based gene signature might serve as a prognostic classifier for clinical decision-making regarding individualized management, follow-up plans, and treatment strategies for ESCC patients.

Pattern of distant metastases and predictive nomograms in colorectal mucinous adenocarcinoma: a SEER analysis.

BACKGROUND: We evaluated the metastatic patterns and explored the prognostic value of distant metastasis pattern in patients with metastatic colorectal mucinous adenocarcinoma (MC) using the Surveillance, Epidemiology, and End Results (SEER) database. METHODS: Between 2010 and 2015, newly diagnosed colorectal MC patients were selected using the SEER database. Patient prognosis was compared based on the clinicopathological parameters, treatment method, and the site and number of metastatic organs. Cox analyses were used to identify factors associated with overall survival (OS). A nomogram was built to predict the patient's survival. Harrell's concordance index (c-index) and calibration curves were used to analyze the discriminative ability of the prognostic factors. RESULTS: Of 3,088 patients diagnosed with colorectal MC, the liver was the only metastatic organ in 78.4% (997/1,271) of all liver metastasis cases, the lung was the only metastatic organ in 41.0% (164/400) of all lung metastasis cases, bone was the only metastatic organ in 26.6% (29/109) of all bone metastasis cases, and the brain was the only metastatic organ in 23.5% (4/17) of all brain metastasis cases. Compared with the untreated cases, those treated with chemotherapy, surgery, and radiotherapy had better OS (P<0.001). There were marked OS differences (P<0.001) between patients with and without liver and bone metastases. Patients with bone metastasis had the best survival, while those with brain metastasis had the worst survival (P<0.001). Patients with one metastatic site had better prognosis compared to those with two or three (P<0.001). Patients with liver metastasis had the best survival, while those with bone and brain metastasis had the worst survival (P<0.001). Multivariate analysis showed that age <65 years, non-black race, grade I, N0 stage, chemotherapy, radiation, surgery, liver metastasis, and bone metastasis were independent prognostic factors. A nomogram was constructed to predict survival probability. The c-index value was up to 0.745. The calibration plot showed that the nomogram was clinically useful. CONCLUSIONS: Metastatic MC (mMC) patients had a characteristic distant metastasis pattern. This study constructed a new and sufficiently accurate prognostic model of mMC based on population-based data. These findings can be utilized to predict prognosis and guide mMC patient management.

Prediction of N7-methylguanosine sites in human RNA based on optimal sequence features.

N-7 methylguanosine (m7G) modification is a ubiquitous post-transcriptional RNA modification which is vital for maintaining RNA function and protein translation. Developing computational tools will help us to easily predict the m7G sites in RNA sequence. In this work, we designed a sequence-based method to identify the modification site in human RNA sequences. At first, several kinds of sequence features were extracted to code m7G and non-m7G samples. Subsequently, we used mRMR, F-score, and Relief to obtain the optimal subset of features which could produce the maximum prediction accuracy. In 10-fold cross-validation, results showed that the highest accuracy is 94.67% achieved by support vector machine (SVM) for identifying m7G sites in human genome. In addition, we examined the performances of other algorithms and found that the SVM-based model outperformed others. The results indicated that the predictor could be a useful tool for studying m7G. A prediction model is available at https://github.com/MapFM/m7g_model.git.

Identification of 2'-O-methylation Site by Investigating Multi-feature Extracting Techniques.

BACKGROUND: RNA methylation is a reversible post-transcriptional modification involving numerous biological processes. Ribose 2'-O-methylation is part of RNA methylation. It has shown that ribose 2'-O-methylation plays an important role in immune recognition and other pathogenesis. OBJECTIVE: We aim to design a computational method to identify 2'-O-methylation. METHODS: Different from the experimental method, we propose a computational workflow to identify the methylation site based on the multi-feature extracting algorithm. RESULTS: With a voting procedure based on 7 best feature-classifier combinations, we achieved Accuracy of 76.5% in 10-fold cross-validation. Furthermore, we optimized features and input the optimized features into SVM. As a result, the AUC reached to 0.813. CONCLUSION: The RNA sample, especially the negative samples, used in this study are more objective and strict, so we obtained more representative results than state-of-arts studies.

Age-related insult of cochlear ribbon synapses: An early-onset contributor to D-galactose-induced aging in mice.

Presbycusis results from age-related degeneration of the auditory system. D-galactose (D-gal)-induced aging is an ideal and commonly used animal model in aging research. Previous studies demonstrate that administration of D-gal can activate mitochondria-dependent apoptosis in the cochlear stria vascularis. However, D-gal-induced changes to cochlear inner (IHCs) and outer (OHCs) hair cells, spiral ganglion cells (SGCs), and ribbon synapses connecting IHCs and SGCs have not been systematically reported. The current study investigated changes in the numbers of hair cells, SGCs, and ribbon synapses in the mouse model of aging. We found that in comparison to control mice, the numbers of ribbon synapses and their nerve fibers were significantly decreased in D-gal-treated mice, whereas the numbers of OHCs, IHCs, and SGCs were almost unchanged. Moreover, hair cell stereocilia were also not obviously influenced by D-gal administration. Although D-gal-induced aging did not significantly shift the auditory brainstem response (ABR) thresholds in the 8, 16, and 32 kHz frequency bands, the amplitude and latency of the ABR wave I, reflecting ribbon synapse functions, were abnormal in D-gal-treated mice compared to control mice. We also found that 8-hydroxy-2-deoxyguanosine, a marker of oxidative DNA damage, was significantly increased in mitochondria of cochleae from mice exposed to D-gal-induced aging in comparison to control mice. Moreover, D-gal administration increased the levels of H2O2 and mitochondrial 3860-bp common deletion, and decreased superoxide dismutase activity and ATP production in the cochlea. Furthermore, compared with control mice, the protein levels of NADPH oxidase 2 and uncoupling protein 2 were significantly increased in the cochlea of D-gal-treated mice. Taken together, these findings support that the cochlear ribbon synapse is the primary insult site in the early stage of presbycusis, and mitochondrial oxidative damage and subsequent dysfunctions might be responsible for this insult.

Mean platelet volume provides beneficial diagnostic and prognostic information for patients with resectable gastric cancer.

Gastric cancer is the fourth most frequent cancer and the second cause of cancer-related mortalities worldwide. Platelets play an important and multifaceted role in cancer progression. Elevated mean platelet volume (MPV) detected in peripheral blood has been identified in various types of cancer. In the present study, we investigated the application value of MPV in early diagnostic and prognostic prediction in patients with resectable gastric cancer. In total, 168 patients with resectable gastric cancer were included and separated into the gastric cancer and healthy control groups according to median pre-operatic MPV value (MPV low, <10.51 or MPV high, ≥10.51). The results showed that the pre-operatic MPV level was significantly higher in gastric cancer patients compared with the healthy subjects. Low pre-operatic MPV level correlated with improved clinicopathological features, including decreased depth of invasion, less lymphonodus metastasis and early tumor stage. The Kaplan-Meier plots showed that the patients with higher pre-operatic MPV had decreased overall survival (OS) and disease-free survival (DFS). Surgical tumor resection resulted in a significant decrease in the MPV level. The patients whose MPV level decreased following surgery had an improved OS. Multivariate Cox regression analysis revealed that the depth of invasion, lymphonodus metastasis, American Joint Committee on Cancer (AJCC) stage, and changes in MPV following surgery were prognostic factors affecting OS, and the AJCC stage and pre-operatic MPV were prognostic factors affecting DFS. In conclusion, MPV measurement can provide important diagnostic and prognostic results in patients with resectable gastric cancer.

Application of platelet/lymphocyte and neutrophil/lymphocyte ratios in early diagnosis and prognostic prediction in patients with resectable gastric cancer.

BACKGROUND: Gastric cancer is the fourth most frequent cancer and the second cause of cancer-related deaths worldwide. China has a high incidence of gastric cancer. Inflammation is a critical component of tumor progression. It has been widely accepted that gastric cancer is an inflammation-driven cancer. In this study, we investigated the application value of systemic inflammatory response (SIR) markers, platelet to lymphocyte ratio (PLR) and neutrophil to lymphocyte ratio (NLR), in early diagnosis and prognostic prediction in patients with resectable gastric cancer. MATERIALS AND METHODS: One hundred and sixty-two patients with resectable gastric cancer were included and separated into groups according to median pre-operative PLR or NLR values (PLR low: < 208 or PLR high: ≥ 208, and NLR low: < 4.02 or NLR high: ≥ 4.02, respectively). To evaluate the changes in PLR or NLR values after operation, we introduced the concept of postpre-operative PLR or NLR ratios (< 1 indicated PLR or NLR values were decreased after operation, while ≥ 1 suggested not decreased PLR or NLR values). RESULTS: Pre-operative PLR and NLR levels were significantly higher in gastric cancer patients compared with the healthy subjects. Low pre-operative PLR and NLR levels correlated with better clinicopathological features, including decreased depth of invasion, less lymph node metastasis and early tumor stage. Kaplan-Meier plots illustrated that higher pre-operative NLR and PLR had decreased overall survival (OS) and disease-free survival (DFS). Surgical tumor resection resulted in a significant CONCLUSIONS: PLR and NLR measurements can provide important diagnostic and prognostic results in patients with resectable gastric cancer.

Changes in neutrophil/lymphocyte and platelet/lymphocyte ratios after chemotherapy correlate with chemotherapy response and prediction of prognosis in patients with unresectable gastric cancer.

The aim of the study was to investigate the application value of neutrophil to lymphocyte ratio (NLR) and the platelet to lymphocyte ratio (PLR) in the prediction of chemotherapy response and prognosis in patients with advanced gastric cancer. In total, 120 patients with unresectable gastric cancer were included and separated into two groups according to the median values of NLR or PLR (NLR low: <4.62 or NLR high: ≥4.62 and PLR low: <235 or PLR high: ≥235, respectively). Low baseline NLR level correlated with improved clinicopathological characteristics, including smaller tumor size, well differentiation and less metastasis. Low baseline PLR level also associated with less metastasis. Patients with a low baseline level of NLR or PLR had an improved response to chemotherapy. Patients with a higher baseline NLR and PLR had decreased progression-free survival (PFS) and overall survival (OS) ratios. Alterations in the NLR and PLR levels were associated with therapeutic efficacy and prognosis. The patients who remained in or switched to the low NLR level subgroup subsequent to first-line chemotherapy had an improved response and improved OS ratios, compared to the patients remaining in or switching to the high NLR level group. Similar results were observed when the PLR level was investigated. In conclusion, baseline NLR and PLR measurements, as well as changes of NLR and PLR following chemotherapy can predict the prognostic results in patients with unresectable gastric cancer.

Mean platelet volume predicts chemotherapy response and prognosis in patients with unresectable gastric cancer.

Gastric cancer is the fourth most frequent cancer and the second cause of cancer-related mortalities worldwide. Platelets play an important and multifaceted role in cancer progression. Elevated mean platelet volume (MPV) detected in peripheral blood has been identified in various types of cancer. In the present study, we investigated the application value of MPV in the prediction of chemotherapy response and prognosis in patients with unresectable gastric cancer. A total of 128 patients with unresectable gastric cancer were included and divided according to the median values of baseline MPV (low MPV: <11.65 or high MPV: ≥11.65). A low baseline MPV level was correlated with reduced metastasis. The results showed that patients with a low baseline level of MPV improved response to chemotherapy. Changes in MPV were associated with therapeutic efficacy. Patients who remained in or were transferred into the low MPV level subgroup following first-line chemotherapy had improved response, compared to those remaining in or being transferred into the high MPV level group. The patients with a higher baseline MPV had decreased progression-free and overall survival ratios. Univariate and multivariate analyses revealed that baseline MPV was a prognostic factor affecting progression-free survival. In conclusion, the results showed that MPV measurements can provide important prognostic information for gastric cancer patients.

[Preparation of chitosan/strontium-substituted hydroxyapatite films on titanium and its FTIR characteristics].

Chitosan/strontium-substituted hydroxyapatite (CHI/SrHAP) coatings were prepared on titanium substrate by electrochemical deposition technique containing Sr2+, Ca2+, PO4(3-) and Chitosan. The as-prepared coatings were examined by scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) tests. The results indicate that the CHI/SrHAP coatings take the morphology of flake-like rather than the needle-like crystal , and the composite coating becomes more compact. The FTIR test indicates that the typical vibration absorption peaks of chitosan (amide I and amide II) emerged, simulated body fluid immersion test proved that the CHI/SrHAP coatings had induced carbonate-apatite formation, indicating that the composite coating possesses excellent biocompatibility. In the electrochemical corrosion testing, that the CHI/SrHAP coatings showed stronger corrosion resistance than pure Ti.

[Preparation of fluoridated hydroxyapatite coatings on titanium by electrolytic deposition and its FTIR characteristics].

Fluoridated hydroxyapatite coatings (FHAP) were prepared on titanium substrate by electrochemical deposition technique containing Ca2+, PO4(3-), and F(-) ions. The deposition was all conducted at a constant current of 0.9 mA for 60 min at 60 degrees C. The as-prepared coatings were examined by scanning electron microscope (SEM), energy-dispersive Xray spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR) and Xray diffraction (XRD) tests. The results indicate that the FHAP cryatals take the morphology of nanoscale-rodlike cone rather than the micron-daisy petal, and the composite coating becomes more compact. The FTIR test indicates that the symmetry of stretching and bending vibration modes of hydroxyl changed, simulated body fluid immersion test proved that the FHAP coating had induced carbonate-apatite formation, indicating that the composite coating possesses excellent biocompatibility.

[Effects of H2 relaxin on airway remodeling and expression of cyclin D1 in a murine model of chronic asthma].

OBJECTIVE: To investigate the effects of H(2) Relaxin (Relaxin) on airway remodeling and the expression of cyclin D(1) in a murine model of chronic asthma. METHODS: Forty BALB/c mice were randomly divided into 4 groups:a normal control group, an asthma group, a vehicle control group and a relaxin treatment group, with 10 mice in each. The mice were sensitized and challenged with ovalbumin (OVA) to establish the chronic asthmatic model. The vehicle control group and the relaxin treatment group were subcutaneously injected with saline and relaxin (0.25 mg × kg(-1)× d(-1))respectively. Alteration of the airway inflammation and collagen deposition were observed by haematoxylin-eosin (HE) and Masson staining. Hydroxyproline in the lung was measured by enzyme linked immunosorbent assay (ELISA). The expression of α-smooth muscle actin (α-SMA) in lungs was evaluated by immunohistochemistry. The protein expression and the mRNA of cyclin D(1) were detected by Western blot and RT-PCR respectively. RESULTS: There were inflammatory cell infiltration, airway stenosis, bronchial smooth muscle hypertrophy and increased collagen deposition in the asthmatic group and the vehicle control group; but these changes were significantly ameliorated in the relaxin treatment group. The area of the α-SMA-stained smooth muscle layer in the asthmatic group and the vehicle control group was significantly greater than that in the control group (all P < 0.05), while administration of relaxin decreased the α-SMA immunostained area (all P < 0.05). The lung hydroxyproline content in the asthmatic and the vehicle groups [(0.68 ± 0.10) mg/g lung tissue, (0.67 ± 0.10) mg/g lung tissue] was significantly greater than that in the control group [(0.26 ± 0.05) mg/g lung tissue] (q = 16.61, 16.01 respectively, all P < 0.01). In contrast, treatment with relaxin significantly reduced the lung hydroxyproline content [(0.40 ± 0.06) mg/g lung tissue] compared with aforementioned 2 groups (q = 10.88, 10.26 respectively, all P < 0.05). The results of the Western blot analysis showed that the expression level of cyclin D(1) in the asthmatic and the vehicle groups [(1.38 ± 0.18), (1.50 ± 0.10)] was higher than that in the control group (0.38 ± 0.10) (q = 13.00, 14.65 respectively, all P < 0.05), while it was significantly decreased in the relaxin group (0.72 ± 0.13) (q = 8.51, 10.16 respectively, all P < 0.05). There were no differences in all of the parameters between the asthmatic group and the vehicle group (P > 0.05). CONCLUSION: Relaxin alleviated airway inflammation, airway smooth muscle thickening and airway remodelling in a murine model of chronic asthma, partially by down-regulating the expression level of cyclin D(1).

[Effects of H2 relaxin on the expression of Epac in a murine model of chronic asthma].

OBJECTIVE: To explore the effects of H(2) relaxin on the expression of exchange protein directly activated by cyclic adenosine monophosphate (Epac) in a murine model of chronic asthma and the roles in the management of airway remodelling. METHODS: Thirty-two BALB/c mice were randomly divided into 4 groups of normal control, asthma, vehicle control and relaxin treatment (n = 8 each). They were sensitized and challenged with ovalbumin to establish a chronic asthmatic model. The vehicle control and relaxin treatment groups were subcutaneously injected with saline and relaxin (0.25 mg×kg(-1)×d(-1)) respectively. Alteration of airway inflammation was observed by hematoxylin-eosin (HE) staining. The airway expressions of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) were evaluated by immunohistochemistry. The protein expression of Epac and phosphorylated extracellular signal regulated kinases1/2 (p-ERK1/2) were detected by Western blot. RESULTS: Compared to those in the normal control group, massive infiltration of inflammatory cells, airway stenosis, bronchial smooth muscle hypertrophy were present in the asthmatic and vehicle control groups. The above-mentioned changes were significantly ameliorated in the relaxin treatment group. The percentage of PCNA positive cells (34.8% ± 6.1%, 33.5% ± 6.6%) and the expression of α-SMA ((1.70 ± 0.25), (1.54 ± 0.24) µm(2)/µm) in the asthmatic and vehicle control groups were significantly higher than those in the normal control group (9.9% ± 2.6%, (0.51 ± 0.16) µm(2)/µm) (all P < 0.05) while administration of relaxin decreased the airway expression levels of PCNA and α-SMA (22.9% ± 5.2%, (1.06 ± 0.25) µm(2)/µm) (all P < 0.05). The results of Western blot showed that the expression levels of Epac in the asthmatic and vehicle groups (0.62 ± 0.12, 0.68 ± 0.11) were lower than those in the control group (1.50 ± 0.17) (all P < 0.05) while it significantly increased in the relaxin group (1.08 ± 0.15) (all P < 0.05). The levels of phosphorylation of ERK1/2 in the asthmatic and vehicle groups (1.45 ± 0.13, 1.36 ± 0.09) were higher than those in the control group (0.38 ± 0.17) (all P < 0.05) while it decreased in the relaxin treatment group (0.72 ± 0.06) (all P < 0.05). No differences existed in all parameters between the asthmatic and vehicle groups (P > 0.05). CONCLUSION: Relaxin alleviates the airway inflammation and airway smooth muscle cell proliferation in a murine model of chronic asthma probably through activating Epac and inhibiting the phosphorylation of ERK1/2.

[Cloning and expression of Hgp44 from Porphyromonas gingivalis].

OBJECTIVE: To clone and express the gene of Hgp44 in adhesin domains of gingipains from Porphyromonas gingivalis (Pg) and to purify the protein. METHODS: The genomic DNA of Pg was isolated from PgATCC33277. The Hgp44 gene fragment was amplified by polymerase chain reaction (PCR) and then inserted into the cloning vector pMD18-T and sequenced. The correct fragment was linked with a prokaryotic expression vector pET-22b to construct the recombinant expression plasmid pET22b-Hgp44. The pET22b-Hgp44 confirmed by enzyme digestion was transformed into competent Escherchia coli (Ec) BL21 (DE3) cells. Expression of fusion protein was induced by isopropyl-ß-D-thiogalactoside (IPTG), and purified by immobilized metal-chelating affinity chromatography (IMAC) using a Ni(2+) matrix column. SDS-polyaerylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis were used to examine the fusion protein. RESULTS: A 1 100 bp fragment was successfully amplified and verified by the agarose gel electrophoresis and sequencing. The generated recombinant expression vectors pET22b-Hgp44 were verified by enzyme digestion and agarose gel electrophoresis. The expression of fusion protein in Ec BL21 (DE3) cells was examined by SDS-PAGE and Western blotting analyses, and the data showed that the protein was 44 000 in size and expressed mostly in the form of inclusion body. The purification of fusion protein was achieved using Ni(2+) affinity chromatography. About 3.5 mg/L fusion protein was obtained. CONCLUSIONS: Hgp44 was successfully expressed in the prokaryotic expression system and purified by IMAC using a Ni(2+) matrix column.