Nardoguaianone L Isolated from Nardostachys jatamansi Improved the Effect of Gemcitabine Chemotherapy via Regulating AGE Signaling Pathway in SW1990 Cells.

Pancreatic cancer is the seventh leading cause of cancer-related death worldwide and is known as "the king of cancers". Currently, gemcitabine (GEM) as the clinical drug of choice for chemotherapy of advanced pancreatic cancer has poor drug sensitivity and ineffective chemotherapy. Nardoguaianone L (G-6) is a novel guaiane-type sesquiterpenoid isolated from Nardostachys jatamansi DC., and it exhibits anti-tumor activity. Based on the newly discovered G-6 with anti-pancreatic cancer activity in our laboratory, this paper aimed to evaluate the potential value of the combination of G-6 and GEM in SW1990 cells, including cell viability, cell apoptosis, colony assay and tandem mass tags (TMT) marker-based proteomic technology. These results showed that G-6 combined with GEM significantly inhibited cell viability, and the effect was more obvious than that with single drug. In addition, the use of TMT marker-based proteomic technology demonstrated that the AGE-RAGE signaling pathway was activated after medication-combination. Furthermore, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) assays were used to validate the proteomic results. Finally, apoptosis was detected by flow cytometry. In conclusion, G-6 combined with GEM induced an increase in ROS level and a decrease in MMP in SW1990 cells through the AGE-RAGE signaling pathway, ultimately leading to apoptosis. G-6 improved the effect of GEM chemotherapy and may be used as a potential combination therapy for pancreatic cancer.

A Carabrane-Type Sesquiterpenolide Carabrone from Carpesium cernuum Inhibits SW1990 Pancreatic Cancer Cells by Inducing Ferroptosis.

Pancreatic cancer has an extremely poor prognosis, and the clinical drugs for the treatment of pancreatic cancer are usually multi-drug combinations. Therefore, it is necessary to search for and find specific new bioactive agents against pancreatic cancer. Carabrone is a carabrane-type sesquiterpenolide extracted from Carpesium cernuum L., and this natural compound has been reported to be a potential anti-tumor agent. However, there are few reports on the function of carabrone related to anti-tumor activity in pancreatic cancer. Herein, cell experiments indicated that carabrone had anti-proliferation inhibition and anti-migration and anti-invasion activity against SW1990 cells. Furthermore, the tandem mass spectrometry and network pharmacology analysis showed that this activity may be related to the ferroptosis and Hippo signaling pathway. Taken together, our results demonstrated that carabrone exhibited prominent anti-pancreatic cancer activity and could be a promising agent against pancreatic cancer.

Prediction of drug-induced eosinophilia adverse effect by using SVM and naïve Bayesian approaches.

Drug-induced eosinophilia is a potentially life-threatening adverse effect; clinical manifestations, eosinophilia-myalgia syndrome, mainly include severe skin eruption, fever, hematologic abnormalities, and organ system dysfunction. Using experimental methods to evaluate drug-induced eosinophilia is very complicated, time-consuming, and costly in the early stage of drug development. Thus, in this investigation, we established computational prediction models of drug-induced eosinophilia using SVM and naïve Bayesian approaches. For the SVM modeling, the overall prediction accuracy for the training set by means of fivefold cross-validation is 91.6 and for the external test set is 82.9 %. For the naïve Bayesian modeling, the overall prediction accuracy for the training set is 92.5 and for the external test set is 85.4 %. Moreover, some molecular descriptors and substructures considered as important for drug-induced eosinophilia were identified. Thus, we hope the prediction models of drug-induced eosinophilia built in this work should be applied to filter early-stage molecules for potential eosinophilia adverse effect, and the selected molecular descriptors and substructures of toxic compounds should be taken into consideration in the design of new candidate drugs to help medicinal chemists rationally select the chemicals with the best prospects to be effective and safe.

[The effect of plasminogen activator inhibitor-1 on embryonic lung fibroblasts from rats].

OBJECTIVE: To explore the effect of plasminogen activator inhibitor-1 (PAI-1) on the proliferation and conversion of rat embryonic lung fibroblasts and the synthesis of collagen, and therefore to explore the function of PAI-1 in pulmonary fibrosis. METHODS: The embryonic lung fibroblasts from pregnant Wistar rats were isolated and cultured in vitro. The reproduction rate of fibroblasts at 12 h, 24 h, and 48 h after being stimulated by PAI-1 with different concentrations (5, 10, 20, 40, 80, and 100 µg/L) was measured by MTT assay. After being stimulated by PAI-1 with the most suitable concentration (20 µg/L) for 48 h and 72 h, the expression of proliferating cell nuclear antigen (PCNA) was measured by immunocytochemical technique, and the mRNA expression of α-SMA and type-1 collagen at 24 h and 48 h was measured by real-time PCR. RESULTS: PAI-1 with different concentrations stimulated the proliferation of fibroblasts. The highest proliferation rate and absorbance in concentration of 20 µg/L and at 12 h were 62.6% and 0.573 ± 0.039. The comparison of different concentrations showed that the difference was significant (F = 111.112, P = 0.000). Therefore, 20 µg/L was selected as the most suitable concentration. Using immunocytochemical method, the optical density of PCNA at 48 h and 72 h were 3685 ± 686 and 2530 ± 477 after being stimulated with 20 µg/L PAI-1. The comparison showed significant difference (F = 7.85, P = 0.02). The expression of α-SMA increased (230 ± 11)% and (159 ± 9)% at 24 h and 48 h after being stimulated with 20 µg/L PAI-1, and the difference was significant (F = 39.92, P = 0.0003). The expression of type-1 collagen increased (92 ± 8)% and (65 ± 12)%, the difference being significant (F = 32.61, P = 0.0006). CONCLUSION: PAI-1 can promote the proliferation and conversion of fibroblasts and the synthesis of collagen, which may be involved in the pathogenesis of pulmonary interstitial fibrosis.

[Effects of leukotriene receptor antagonists on vascular endothelial growth factor and its receptors in a sensitized rat model].

OBJECTIVE: To investigate the effect of montelukast (MK) on airway inflammation and remodeling in asthmatic rats, and to explore the regulating role of MK on vascular endothelial growth factor (VEGF) and its receptors. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into 3 groups, a control group (n = 8), an asthmatic group (n = 8) and a MK treated group (n = 8). The rats were sensitized with ovalbumin and AL (OH3), and repeatedly exposed to aerosolized ovalbumin. Airway reactivity of the animals were measured by animal lung function meter. VEGF levels and leukotriene D(4) (LTD(4)) in serum were measured by enzyme linked-immunosorbent assay (ELISA). The pathologic changes of bronchi and the lung tissue were evaluated, and the expression of VEGF and its acceptors was analyzed with immunohistochemistry. The vascular counts and vascular smooth muscle thickness were measured by using image analysis system. RESULTS: The bronchial provocation test showed that, in the asthmatic group, the average expiratory resistance increased remarkably. The serum levels of VEGF and LTD(4) in the asthmatic group were 31 +/- 6 and 11 +/- 4 respectively, significantly higher than those in the control group (17 +/- 5 and 6.1 +/- 0.7) respectively and in the MK group (15 +/- 4 and 9.8 +/- 1.6) respectively. (F 63.78, 39.56 all P < 0.01). Immunohistochemistry showed that, the expression of VEGF, VEGFR(1) and VEGFR(2) in the asthmatic group were increased, as compared to those in the control group and the treated group. The vascular counts were 14 +/- 2, 22 +/- 2 and 16 +/- 4 in the control, the asthmatic, and the treated groups. CONCLUSIONS: VEGF and its receptors were over-expressed in the sensitized rat model, and involved in angiogenesis and airway remodeling. MK may be effective in reducing allergic airway inflammation and airway remodeling through VEGF and VEGFR.

[The change and significance of the cardiopulmonary exercise test parameters in patients with obstructive sleep apnea-hypopnea syndrome].

OBJECTIVE: To investigate the change of exercise cardiopulmonary function in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). METHODS: Thirty OSAHS patients and 18 normal healthy adults (control group) were studied by cardiopulmonary exercise test (CPET). The results including maximal oxygen uptake percent predicted (Vo(2)max% predicted), oxygen uptake to work rate (Vo(2)/WR), oxygen pulse percent predicted (Vo(2)/HRmax% predicted), anaerobic threshold to maximal oxygen uptake (AT/Vo(2)max), breathing reserve (V(E)max/MVV) and ventilatory equivalents for carbon dioxide (V(E)/V(CO2)) were compared between two groups. RESULTS: The levels of Vo(2)max% predicted, AT/Vo(2)max, Vo(2)/HRmax% predicted, Vo(2)/WR, and V(E)max/MVV in the OSAHS group [(83 +/- 5)%, (44 +/- 6)%, (79 +/- 5)%, (9.3 +/- 0.6) ml.min(-1).W(-1), (73 +/- 8)%] were lower than those in the control group [(88 +/- 5)%, (49 +/- 6)%, (83 +/- 4)%, (10.9 +/- 2.3) ml.min(-1).W(-1), (79 +/- 9)%, all P < 0.05]. The levels of V(E)/V(CO2) in the OSAHS group (29 +/- 3) was higher than the control group (26 +/- 3, P < 0.05). In the OSAHS group Vo(2)max% predicted, Vo(2)/HRmax% predicted, Vo(2)/WR AT/Vo(2)max and V(E)max/MVV correlated negatively with apnea-hypopnea index (AHI, r = -0.52, -0.62, -0.59, -0.37, -0.66, P < 0.05). Vo(2)max% predicted, Vo(2)/HRmax% predicted, Vo(2)/WR, AT/Vo(2)max and V(E)max/MVV correlated with lowest oxygen saturation (LSaO(2), r = 0.60, 0.63, 0.64, 0.40, 0.59, P < 0.05). V(E)/V(CO2) correlated with AHI (r = 0.57, P < 0.01) and correlated negatively with LSaO(2) (r = -0.62, P < 0.01). CONCLUSIONS: The cardiac output of patients with OSAHS can not meet the demand of hard exercise. At the same time, there is more significant ventilation-perfusion disturbance in OSAHS patients than normal subjects. The patients' exercise cardiopulmonary function has been compromised although there are no symptoms.

[Changes of coagulation and fibrinolysis system in bronchoalveolar lavage fluid in lung fibrosis].

OBJECTIVE: To investigate the activity of coagulation and fibrinolysis systems in bronchoalveolar lavage fluid (BALF) in lung fibrosis in rats, and to study their role in diagnosis of lung fibrosis. METHODS: Thirty-six Sprague-Dawley rats were randomly divided into 2 groups (n=18 in each group ). Lung fibrosis and control models were made by tracheal instillation bleomycinA(5) (BLMA(5)) (5 mg/kg) and saline. On days 7, 14 and 28, the recalcification time points of normal pooled plasma for studying procoagulation activity (PCA), the levels of von Willebrand Factor (vWF), the activities of Antithrombin-III(AT-III) , plasminogen activity inhibitor-1 (PAI-1) and urokanise-type plasminogen activator (uPA) in BALF were measured respectively. RESULTS: (1) In BLM group,the recalcification time points in BALF were (56+/-10), (78+/-3) and (172+/-11) seconds respectively, they were (190+/-10), (186+/-8) and (184+/-6) seconds respectively in control group. The difference was significant (P<0.01) except on day 28; (2) The differences in level of vWF and activity of AT-III in BALF were not significant in BLM group versus control group; (3) Activities of PAI-1 in BALF in BLM group were (1.04+/-0.08), (1.47+/-0.06), (3.03+/-0.18) u/mL respectively, while they were (0.66+/-0.11), (0.71+/-0.09), (0.70+/-0.08) u/mL respectively in control group. The difference was significant (P<0.01); (4) Activities of uPA in BALF in BLM group were (0.19+/-0.03), (0.16+/-0.02), (0.12+/-0.05) u/mL respectively, while they were (0.26+/-0.05), (0.25+/-0.06), (0.24+/- 0.07) u/mL respectively. The difference was significant (P<0.01). CONCLUSION: The PCA upregulation might be means to diagnose alveolitis. The activity of the fibrinolysis system was injured progressively by PAI-1 activity upregulation and uPA activity downregulation, which might be taken as means to study the occurrence of early lung fibrosis and its development,vWF and AT-III rendered no help in diagnosis of lung fibrosis.

[The change and significance of coagulation activity in bleomycin-induced lung fibrosis in rats].

OBJECTIVE: To investigate the role of coagulation activity of bronchoalveolar lavage fluid (BALF) in the pathogenesis of lung fibrosis. METHODS: Fourty-eight Sprague-Dawley rats were randomly divided into 2 groups, 24 rats in each group. In the bleomycin (BLM) group, the lung fibrosis model was made by tracheal instillation of bleomycin A(5) (BLMA(5), 5 mg/kg). At day 7, 14, 28 and 40, the recalcification time of normal pooled plasma, factor VII and X deficiency plasma were measured for procoagulation activity (PCA), and the thrombin activity and the protein level of transforming growth factor beta(1) (TGF-beta(1)) in BALF were also measured. In the control group, normal solution was instillated into the lungs. RESULTS: In the BLM group, the recalcification time of normal pooled plasma in BALF at the four time points were (56 +/- 10), (78 +/- 4), (172 +/- 11) and (180 +/- 6) s respectively, while in the control group, were (190 +/- 10), (186 +/- 8), (184 +/- 6) and (185 +/- 6) s respectively. The thrombin activity at the four time points were (1.26 +/- 0.03), (0.82 +/- 0.05), (0.28 +/- 0.03) and (0.28 +/- 0.02) microg/ml respectively in the BLM group, but were (0.31 +/- 0.02), (0.32 +/- 0.03), (0.31 +/- 0.04) and (0.29 +/- 0.05) microg/ml respectively in the control group. The level of TGF-beta(1) at the four time points were (310 +/- 36), (220 +/- 30), (109 +/- 12) and (96 +/- 11) ng/ml respectively in the BLM group, but were (92 +/- 20), (94 +/- 12), (92 +/- 10) and (90 +/- 9) ng/ml respectively in the control group. The above measurements were significantly different in day 7 and 14 between the BLM group and the control group (P < 0.01), while the differences were not significant at day 28 and 40 (P > 0.01). In the BLM group, at day 7 and 14, the recalcification time of factor VII deficiency plasma was (123 +/- 12) and (162 +/- 4) s respectively; the recalcification time of factor X deficiency plasma was (357 +/- 22) and (387 +/- 12) s respectively; the recalcification time of factor X deficiency plasma was longer than that of factor VII deficiency plasma and that of normal pooled plasma. Within 14 day, the level of TGF-beta(1) was positively correlated with PCA and thrombin activity. CONCLUSIONS: During the period of alveolitis, the PCA and thrombin activity were upregulated in BALF, which was caused by activated factor VII activating factor X and the switching to the exogenous coagulation pathway. But during the period of lung fibrosis, their activities were not upregulated. These results suggest that the coagulation factor and thrombin might contribute to the development of pulmonary fibrosis by promoting production of TGF-beta(1).

[Inhibitory effect of hTERT dsRNA on telomerase activity in lung carcinoma cell line A549].

BACKGROUND & OBJECTIVE: RNA interference (RNAi) is a new technology in gene study. The mechanism of RNAi is that double-stranded RNA (dsRNA) can band target mRNA and decompose it. This study was to assess possibility and specificity of dsRNA on suppressing human telomerase reverse transcriptase (hTERT) in lung carcinoma cells, investigate its effect on cell proliferation to confirm whether it has unspecific killing activity on mammalian cells, and explore its application in lung cancer research and treatment. METHODS: Sequences of 2 exons and 1 intron of hTERT gene were amplified by reverse transcription-polymerase chain reaction (RT-PCR) or PCR. The sense and antisense cDNA sequences were connected in a tandem manner, and the whole fragment was inserted into pCI-neo mammalian expression vector to construct the dsRNA expression vector, and then transfected into lung carcinoma cell line A549. The expression of hTERT was detected by RT-PCR and Western blot. Telomerase activity was measured by telomerase repeat amplification protocol (TRAP). Cell morphology was observed, and cell proliferation was assessed under invert microscope. RESULTS: After transfection of 2 exon fragments of hTERT dsRNA, mRNA and protein expression of hTERT and telomerase activity in A549 cells were suppressed, cell proliferation was markedly inhibited. Meanwhile, dsRNA didn't show unspecific toxic activity on A549 cells. CONCLUSIONS: hTERT dsRNA can specifically silent hTERT gene, inhibit telomerase activity and proliferation of A549 cells. hTERT dsRNA might be a potential method of gene therapy for lung cancer.

[Study on hemodynamic changes and cardiac troponin I level in pig model of acute experimental pulmonary embolism].

OBJECTIVE: To evaluate the hemodynamic effects and cardiac troponin I (cTn I), creatine kinase-MB (CK-MB), myoglobin (Mb) releasing kinetics of acute experimental pulmonary embolism of pigs. METHODS: Sixteen juvenile pigs, of either gender and weighing 30 to 40 kg were studied, 8 in the embolism group and 8 in the control group. The 8 embolism animals received 0.1 g/kg polystyrene beads (diameter range 0.65 to 0.67 mm) suspended in 0.9% saline by venous injection. Pulmonary arterial pressure (PAP), systemic arterial pressure (SAP), pulmonary capillary wedged pressure (PCWP), cardiac output (CO), blood gases and serum cTn I, CK-MB, and Mb were measured before and immediately, 30 min, 1 hour, 2 hour, and 3 hour after acute pulmonary embolism. RESULTS: PAP was increased to 2 - 3 fold of the baseline and the control level immediately, and then decreased to the baseline level in 2 to 3 hours. Serum cTn I and Mb increased significantly after embolism and remained at a higher level through the 3 hour experimental procedure. The CK-MB was not changed after acute pulmonary embolism. CONCLUSIONS: Acute pulmonary embolism caused lung gas exchange abnormality and acute pulmonary hypertension. The hemodynamic effects of acute pulmonary embolism include injury to the myocardial cells and releasing of cTn I and Mb to blood stream. cTn I can be detected in the early phase of acute pulmonary embolism, and maybe a useful marker in diagnosis and management of acute pulmonary embolism.

[Inhibition of anti-sense human telomerase reverse transcriptase (hTERT) retroviral vector on lung cancer cells].

BACKGROUND & OBJECTIVE: Inhibition of telomere length can be achieved through suppression of telomerase activity, which may result in the inhibition of immortal cell proliferation. In order to explore the possibility of the telomerase as a target for lung cancer therapy, we investigated the effects of anti-sense human telomerase reverse transcriptase (hTERT) on telomerase activity and cell proliferation of A549 lung cancer cell line. METHODS: The anti-sense hTERT cDNA, an 835 bp in the 5' region of hTERT mRNA was amplified by reverse transcription polymerase chain reaction (RT-PCR), before cloning into pLXSN retroviral vector in sense and anti-sense orientations. A549 cells, a human lung cancer cell line, were infected with recombinant virus obtained after transfection into packaging cell PT67. The expression of hTERT protein was determined by Western blot analysis. The telomerase activity was measured by telomerase repeat amplification protocol (TRAP). The cell proliferation was depicted by cell morphology under inverted microscopy as well as cell growth curve. Apoptosis was analyzed by flow cytometry and DNA electrophoresis. RESULTS: Compared with sense hTERT transduction, hTERT expression and telomerase activity significantly decreased in A549 cells after anti-sense hTERT transduction. The cell proliferation was markedly inhibited with evidence of apoptosis. CONCLUSION: Anti-sense hTERT exhibited significant inhibition of telomerase activity and cell proliferation, in addition to acceleration of apoptosis. This implied the possibility of hTERT as the potential target for gene therapy of lung cancer.

The involvement of glia in long-term plasticity in the spinal dorsal horn of the rat.

We have examined the potential role of spinal glial cells in the induction of C fiber-evoked long-term potentiation (LTP) in the spinal cord. Tetanic stimulation of the sciatic nerve induced longterm potentiation of C-fiber-evoked field potentials in the spinal dorsal horn in all rats. Following intrathecal fluorocitrate (1 nmol), a glial metabolic inhibitor, tetanic stimulation induced longterm depression (LTD) but not LTP. The effects of fluorocitrate were abolished by kynurenic acid or 2-amino-5-phosphonovaleric acid (AP-5), but not by 6,7-dinitroquinoxaline-2,3-dione (DNQX), picrotoxin or strychnine. These data suggest that spinal glial cells may modulate the central sensitization of nociceptive neurons via NMDA receptors.

Constrictive pericarditis presenting as massive ascites in children: report of one case.

Abdominal distension has been described as the most common presenting symptom in children with constrictive pericarditis. This report describes a 13-year-old boy who had abdominal distension with massive ascite and hepatosplenomegaly as an initial presentation. The physical signs of jugular vein engorgement and gallop rhythm as well as the pericardial calcification on the chest roentgenogram lead to the diagnosis of constrictive pericarditis. After ultrafast computed tomography and cardiac catheterization confirmation, the patient received a pericardiectomy with excellent relief of symptoms. Pathology of the pericardium reveals fibrocalcified change, but no acid fast stained bacillus nor granulomatous lesion was observed. The incidence of constrictive pericarditis with evident pericardial calcification in children is extremely low. The diagnostic value of the chest roentgenogram and physical findings for the constrictive pericarditis are addressed.