Polyphyllin B inhibited STAT3/NCOA4 pathway and restored gut microbiota to ameliorate lung tissue injury in cigarette smoke-induced mice.

OBJECTIVE: Smoking was a major risk factor for chronic obstructive pulmonary disease (COPD). This study plan to explore the mechanism of Polyphyllin B in lung injury induced by cigarette smoke (CSE) in COPD. METHODS: Network pharmacology and molecular docking were applied to analyze the potential binding targets for Polyphyllin B and COPD. Commercial unfiltered CSE and LPS were used to construct BEAS-2B cell injury in vitro and COPD mouse models in vivo, respectively, which were treated with Polyphyllin B or fecal microbiota transplantation (FMT). CCK8, LDH and calcein-AM were used to detect the cell proliferation, LDH level and labile iron pool. Lung histopathology, Fe3+ deposition and mitochondrial morphology were observed by hematoxylin-eosin, Prussian blue staining and transmission electron microscope, respectively. ELISA was used to measure inflammation and oxidative stress levels in cells and lung tissues. Immunohistochemistry and immunofluorescence were applied to analyze the 4-HNE, LC3 and Ferritin expression. RT-qPCR was used to detect the expression of FcRn, pIgR, STAT3 and NCOA4. Western blot was used to detect the expression of Ferritin, p-STAT3/STAT3, NCOA4, GPX4, TLR2, TLR4 and P65 proteins. 16S rRNA gene sequencing was applied to detect the gut microbiota. RESULTS: Polyphyllin B had a good binding affinity with STAT3 protein, which as a target gene in COPD. Polyphyllin B inhibited CS-induced oxidative stress, inflammation, mitochondrial damage, and ferritinophagy in COPD mice. 16S rRNA sequencing and FMT confirmed that Akkermansia and Escherichia_Shigella might be the potential microbiota for Polyphyllin B and FMT to improve CSE and LPS-induced COPD, which were exhausted by the antibiotics in C + L and C + L + P mice. CSE and LPS induced the decrease of cell viability and the ferritin and LC3 expression, and the increase of NCOA4 and p-STAT3 expression in BEAS-2B cells, which were inhibited by Polyphyllin B. Polyphyllin B promoted ferritin and LC3II/I expression, and inhibited p-STAT3 and NCOA4 expression in CSE + LPS-induced BEAS-2B cells. CONCLUSION: Polyphyllin B improved gut microbiota disorder and inhibited STAT3/NCOA4 pathway to ameliorate lung tissue injury in CSE and LPS-induced mice.

Repeat-dose toxicity study of human umbilical cord mesenchymal stem cells in cynomolgus monkeys by intravenous and subcutaneous injection.

Human umbilical cord mesenchymal stem cells (hUC-MSCs) are proposed for the treatment of acute lung injury and atopic dermatitis. To advance hUC-MSC entry into clinical trials, the effects of hUC-MSCs on the general toxicity, immune perturbation and toxicokinetic study of hUC-MSCs in cynomolgus monkeys were assessed. hUC-MSCs were administered to cynomolgus monkeys by intravenous infusion of 3.0 × 106 or 3.0 × 107cells/kg or by subcutaneous injection of 3.0 × 107cells/kg twice a week for 3 weeks followed by withdrawal and observation for 6 weeks. Toxicity was assessed by clinical observation, clinical pathology, ophthalmology, immunotoxicology and histopathology. Moreover, toxicokinetic study was performed using a validated qPCR method after the first and last dose. After 3rd or 4th dosing, one or three the monkeys in the intravenous high-dose group exhibited transient coma, which was eliminated by slow-speed infusion after 5th or 6th dosing. In all dose groups, hUC-MSCs significantly increased NEUT levels and decreased LYMPH and CD3+ levels, which are related to the immunosuppressive effect of hUC-MSCs. Subcutaneous nodules and granulomatous foci were found at the site of administration in all monkeys in the subcutaneous injection group. Other than above abnormalities, no obvious systemic toxicity was observed in any group. The hUC-MSCs was detectable in blood only within 1 h after intravenous and subcutaneous administration. The present study declared the preliminary safety of hUC-MSCs, but close monitoring of hUC-MSCs for adverse effects, such as coma induced by intravenous infusion, is warranted in future clinical trials.

MiroRNA-31-3p Promotes the Invasion and Metastasis of Non-Small-Cell Lung Cancer Cells by Targeting Forkhead Box 1 (FOXO1).

OBJECTIVE: To explore the possibility of microRNA miR-31-3p as a biomarker for bone metastasis of non-small-cell lung cancer (NSCLC) and its molecular mechanism to the invasion and metastasis of NSCLC cells. METHODS: Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of miR-31-3p and forkhead box 1 (FOXO1) in NSCLC tissues, serum, and cells to analyze the correlation between the expression levels of miR-31-3p and the clinicopathology of NSCLC. After interference with or overexpressing miR-31-3p, NSCLC cell proliferation, apoptosis, invasion ability, and migration ability were detected by MTT, flow cytometry, Transwell, and scratch experiment, respectively. The interaction between miR-31-3p and FOXO1 was further verified by the dual-luciferase reporter experiment. Western blot was performed to detect the protein expression of FOXO1 in tissues and FOXO1, RhoA, p-RhoA, ROCK-2, and p-ROCK-2 in cells. RESULTS: In tissues, serum, and NSCLC cell line A549 of the NSCLC patients, the expression of FOXO1 was notably lower, and the miR-31-3p expression was significantly higher. Overexpression of miR-31-3p could distinctly improve the proliferation, invasion, and migration of A549 cells, meanwhile inhibit cell apoptosis, and activate the RhoA/ROCK-2 signaling pathway, while interfering with the expression of miR-31-3p has the opposite function. Besides, bioinformatics analysis and luciferase reporter assay confirmed that FOXO1 was a target gene of miR-31-3p. Overexpressing FOXO1 could inhibit the proliferation and metastasis of A549 cells, but overexpressing miR-31-3p reverses the results. CONCLUSION: This study confirmed that miR-31-3p promotes the proliferation, invasion, and migration of NSCLC cells and inhibits apoptosis through targeted regulating FOXO1 and be a potential therapeutic targets for the treatment of NSCLC.

Long non-coding RNA MALAT1 sponges miR-149 to promote inflammatory responses of LPS-induced acute lung injury by targeting MyD88.

Acute lung injury (ALI) caused by sepsis occurs early and the condition is severe, and is also an important reason for accelerating the death of patients. Increasing evidence has identified long non-coding RNA (lncRNA) metastasis associated in lung adenocarcinoma transcript 1 (MALAT1) as a regulator of ALI. However, the potential mechanism underlying MALAT1 on ALI still needs further identification. To explore the mechanisms of gene regulation expression mediated by MALAT1 through miR-149/MyD88 in lung injury inflammation, we constructed a lung injury inflammatory model using the lipopolysaccharides (LPS)-induced method and quantificated the cytokines and signaling cascade molecules as well as miR-149. The MALAT1, myeloid differentiation factor 88 (MyD88), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and IL-6 levels were significantly increased, and the nuclear factor-κB (NF-κB) pathway was activated, but the miR-149 level was decreased in the LPS-induced ALI model. miR-149 directly targeted both lncRNA MALAT1 and the MyD88 gene. Knockdown of MALAT1 down-regulated the levels of MyD88, TNF-α, IL-1ß, and IL-6, and inhibited the NF-κB pathway. However, MALAT1 knockdown up-regulated the expression of miR-149. Overexpression of miR-149 down-regulated MyD88, TNF-α, IL-1ß, and IL-6 levels, and inhibited the NF-κB pathway. MALAT1 acts as a pro-inflammatory factor in ALI via the miR-149/MyD88/NF-κB axis and is therefore a potential novel therapeutic target for ALI treatment.

Krüppel-Like Factor 4 Enhances Sensitivity of Cisplatin to Lung Cancer Cells and Inhibits Regulating Epithelial-to-Mesenchymal Transition.

In order to improve therapeutic efficacy, it is a current emergency to better know the mechanisms underlying cisplatin resistance in lung cancer cells. In this study, we aim to investigate the role of Krüppel-like factor 4 (KLF4) in cisplatin-resistant lung cancer cells. We developed cisplatin-resistant lung cancer cell line A549/DDP, and then a battery of experiments was used to analyze the effects of KLF4 in cisplatin resistance of lung cancer. We found that KLF4 was significantly downregulated in cisplatin-resistant A549 cells and forced KLF4 expression inhibited cell growth and induced apoptosis. Further, we found that overexpression of KLF4 was able to inhibit cell migration and invasion, to inhibit the expression of Slug, Twist, and vimentin, and to increase the expression of E-cadherin and subsequent inhibition of the EMT process. Thus, overexpression of KLF4 may be a potential strategy for lung cancer treatment, especially for cisplatin-resistant cases.

[Clinical efficacy of vitamin support in lung adenocarcinoma patients treated with pemetrexed second-line chemotherapy].

OBJECTIVE: To analyze the clinical efficacy and toxicity of vitamin support in lung adenocarcinoma patients treated with pemetrexed second-line chemotherapy. METHODS: Two hundred and eighty-three patients with stage 3/4 lung adenocarcinoma treated at our hospital from August 2010 to August 2013 were included in this study. The lung adenocarcinomas in all the 283 patients were confirmed by pathology or cytology, all were EGFR-negative, and all patients received pemetrexed second line chemotherapy. The 283 patients were randomly divided into two groups: the improved treatment group (142 cases) and the conventional treatment group (141 cases). The patients of conventional treatment group received 400 µg folic acid per os daily for 7 days before the first dose of pemetrexed, and continued until 21 days after the last dose of pemetrexed. Besides, they received 1000 µg vitamin B12 injection at 7 days before the first dose of pemetrexed, and once per cycle of pemetrexed for 3 cycles after the last dose of pemetrexed. The patients of the improved treatment group took 400 µg folic acid daily per os from the day before the first dose to 21 days after the last dose of pemetrexed. They also received 500 µg vitamin B12 by injection one day before the first dose, and one day before each therapy cycle of pemetrexed therapy. RESULTS: The mean number of cycles of pemetrexed chemotherapy was 4 in both groups. In the 142 patients of improved treatment group, complete response (CR) was observed in two cases, partial remission (PR) in 28, stable disease (SD) in 21, and progressive disease (PD) in 91 cases, with a total effective rate of 21.1%. While in the conventional treatment group, CR was observed in one case, PR in 27 cases, SD in 23 cases, and PD in 90 cases, with a total effective rate of 19.9%. The median progression-free survival (PFS) was 3.8 months in the improved treatment group and 4.2 months in the conventional treatment group (P=0.143). The toxicity of chemotherapy was mild in both groups, with no significant difference between the two groups (P>0.05). The most common side effects of hematological system were leukopenia and neutropenia, and the most common side effects of non-blood system were nausea and vomiting. The most common grade 3-4 toxic reaction in both groups was leukopenia and neutropenia, with no significant difference between the two groups (P>0.05). Multivariate analysis showed that the age of patients was an independent factor of grade 3-4 chemotherapy toxic reaction (P<0.05), while gender, the baseline level of PS score or blood system had no significant effect on the grade 3-4 chemotherapy toxic reaction (P>0.05). CONCLUSIONS: Compared with the conventional treatment scheme, the improved treatment scheme has similar therapeutic effects and could be used more conveniently, while the toxic effects of chemotherapy are not increased at the same time. Our results indicate that pemetrexed-based chemotherapy does not need to delay the chemotherapy because of vitamin support treatment.

[The diagnostic value of flexible bronchoscopy in tracheobronchopathia osteochondroplastica].

OBJECTIVE: To enhance the knowledge of tracheobronchopathia osteochondroplastica (TO), and to describe the value of flexible bronchoscopic diagnosis and treatment for the disease. METHODS: The clinical data, bronchoscopic findings, histological results and the methods and effect of bronchoscopic treatment in 10 patients with TO admitted to Xiangya Hospital between June 2006 and July 2007 were retrospectively analyzed. RESULTS: There were 8 males and 2 females (mean age 46 +/- 16, range 33-76 years). The bronchoscopic appearance of TO was multiple whitish, hard nodules projecting into the tracheal lumen (mostly from the anterior and less from the lateral walls). The lesions were found most frequently in the trachea and major bronchi, and lobar and segmental bronchi were involved less frequently. Nodules were restricted to the anterolateral walls in 7 cases. The distribution of the lesions was diffuse in 5, confluent in 2 and scattered in 3 cases. Six patients received bronchoscopic management, including radiofrequency treatment for 2 patients and argon ion laser treatment for the other 4. The lesions in the airways were reduced and clinical symptoms improved to some extent after treatment. No severe complications occurred during and after the procedures. CONCLUSIONS: The diagnosis of TO can be easily underdiagnosed or misdiagnosed. Flexible bronchoscopy with histological examination is the main method for the diagnosis of TO. Radiofrequency and argon ion laser treatment are safe and effective.

[Bronchoscopic argon plasma coagulation therapy for bronchial carcinoma].

OBJECTIVE: To evaluate the effect of bronchoscopic argon plasma coagulation therapy on bronchial carcinoma. METHODS: Thirty-one bronchial carcinoma patients were diagnosed by bronchoscope and pathological tests, with or without atelectasis or obstructive pneumonia on chest X-ray or chest CT. Argon plasma coagulation therapy was performed through bronchoscope. The location of the airway lesions, the degree of obstruction, dyspnea index, and complications were evaluated. RESULTS: The patients with bronchial carcinoma were treated 1-4 times by bronchoscopic argon plasma coagulation therapy. Full effectiveness was achieved in 15 patients (48.4 %), partial in 12 (38.7 %), and mild in the other 4 (12.9 %). The overall effective rate was 100%. CONCLUSION: Bronchoscopic argon plasma coagulation therapy for bronchial carcinoma can remarkably reduce the tumor size, relieve clinical symptoms, and alleviate the obstruction caused by bronchial neoplasm. Bronchoscopic argon plasma coagulation therapy is an effective and safe method for patients with bronchial carcinoma.

Development of a rat model by 3,4-benzopyrene intra-pulmonary injection and evaluation of the effect of green tea drinking on p53 and bcl-2 expression in lung carcinoma.

BACKGROUND: A convenient animal model of primary lung cancer is compelling for investigation into the disease mechanisms and for development of therapeutic strategies. This study aims to develop a reproducible rat model for lung carcinoma by intra-pulmonary injection of 3,4-benzopyrene, and to evaluate the preventive effect of green tea on the formation of lung carcinoma. METHODS: Sprague-Dawley rats of the same ages were randomly assigned into three groups treated differently. Rats in group one were given green tea in drinking water (tea concentration: 1.2%; tea polyphenols in the tea solution: 0.3%); rats in the groups two and three were given blank drinking water. Rats in the groups one and two were injected intra-pulmonarily with 3,4-benzopyrene dissolved in corn oil (2 mg/0.2 mL/injection, fortnightly, 4 times in all); rats in the group three were injected with the vehicle corn oil as the control for injection. All the rats were sacrificed one year after the first intra-pulmonary injection. Tumors developed in rats and lung tissues were collected for carcinoma diagnosis and for p53 and bcl-2 expression. RESULTS: Intra-pulmonary injection of 3,4-benzopyrene steady induced lung carcinoma at a success rate of 75%. Administration with green tea drinking significantly reduced the incidence of lung carcinoma to 30%. Green tea up-regulated p53 expression in lung carcinoma, but significantly down-regulated bcl-2 expression. CONCLUSIONS: Intra-pulmonary injection of 3,4-benzopyrene can steady induce lung carcinoma in rats, and green tea has preventive effect against lung cancer possibly by regulating expression of some critical genes such as p53 and bcl-2.

[Analysis of bronchoscopy characteristics for 729 female patients with lung cancer.].

BACKGROUND: The aim of this article is to study features of the bronchoscopy signs in female lung cancer patients. METHODS: The bronchoscopy data of 729 female lung cancer patients enrolled between January 1994 and June 2007 was analyzed, retrospectively. RESULTS: Most of the patients were middle-aged female (57.0%), then were the elderly (28.5%), and the youth composed much lower (14.0%). The most common histopathology was adenocarcinoma (42.8%), followed by squamous cell carcinoma (23.9%) and small cell carcinoma (19.2%), and all of them increased in the past few years. The female lung cancers were more in the right lung (P <0.05), and the upper lobes (P <0.05). Among 729 female lung cancer patients, 92.0% had apparent signs. Most of adenocarcinoma had infiltrative changes (P <0.05), but most of squamous cell carcinoma and small cell carcinoma had proliferative changes (P <0.05). The most common sing of bronchoscopy in patients with atelectasis was proliferative changes (P <0.05), but the most common sings of bronchoscopy in patients with pleural effusion was infiltrative changes (P <0.05). CONCLUSIONS: This study suggests brochoscopy is an important approach in diagnosis of female lung cancer. Paying more attention to the lung cancer of female patients and examining with bronchoscopy would be helpful for earlier diagnosis.

[Prevention of chinese green tea on 3,4-benzopyrene-induced lung cancer and its mechanism in animal mode.].

BACKGROUND: Chinese green tea is one of the daily consumption beverages in the world and is considered a promising cancer chemopreventive agent. In the present study, we investigate the role of lung cancer prevention by green tea and its mechanism. METHODS: Three groups of female SD rats were kept with the same feed. Rats in group A were administrated with 1% green tea drinking, while in group B and group C with water only. Animals in group A and group B were given 3,4-benzopyrene-corn oil mixture pulmonary injection fortnightly for 4 times, while in group C corn oil only. Rats were sacrificed 1 year after the first injection under narcotism. Lung tumors and lung tissues were performed H&E staining for cancer identification. Each case of lung cancer was examined for expression of p53 and Bcl-2 with in situ hybridization analysis and immunohistochemistry staining. RESULTS: No cancer was found in rats in group C. However, in group B, 15 out of 20 rats were found generating lung cancer, and in group A, 6 out of 20 rats inducing lung cancer were recorded. The rate of lung carcinogenesis in rats was decreased from 75% to 30% by 1% chinese green tea oral administration (Chi-Square=8.12, P <0.01). Higher level of p53 expression in lung cancer tissues of group A was observed under microscope than that of group B, but the difference has no statistic significance (P >0.05). However, significantly lower level of Bcl-2 expression was found in lung cancer tissues of group A than that of group B (P <0.05). CONCLUSIONS: The results indicate that chinese green tea inhibits lung carcinogenesis. Chinese green tea can slightly upregulate expression of p53, but significantly downregulate expression of Bcl-2 in lung cancer, and this may be related to the mechanism of lung cancer prevention.

[Effect of ginsenoside Rh2 on immunocompetence of alveolar macrophages in patients with non-small cell lung cancer].

OBJECTIVE: To explore the effect of ginsenoside Rh2 (G-Rh2) on the excretion of cytotoxin-effecting molecule of alveolar macrophages (AM) in patients with non-small cell lung cancer (NSCLC). METHODS: The concentration of tumor necrosis factor (TNF-alpha) and NO in the bronchoalveolar lavage fluid (BALF) and the cultured supernatants of AM in 35 patients with NSCLC were measured by ELISA and enzyme method,and levels of TNF-alpha and NO in the cultured supernatants of AM after being cultivated with IFN-alpha, G-Rh2, and IFN-alpha+G-Rh2 were measured by the same method. RESULTS: AM in all the non-small cell lung cancer patients produced TNF-alpha and NO. The activity of TNF-alpha and NO was lower in the BALF and in the cultured supernatants of AM of the tumor-bearing lungs than that of the non-tumor-bearing lungs. The concentrations of TNF-alpha and NO in the cultured supernatants of AM cultivated with G-Rh2 were higher than those in the control (P<0.05), but there were no significant differences between the G-Rh2 group and IFN-alpha group (P>0.05). The concentrations of TNF-alpha and NO in the cultured supernatants of AM cultivated with both G-Rh2 and IFNalpha were obviously higher than those stimulated with IFNalpha or G-Rh2 (P<0.01) alone. CONCLUSION: G-Rh2 can enhance the excretion of cytotoxin-effecting molecules of AM in patients with NSCLC. The changes are more distinctive when G-Rh2 and IFNalpha have coordinated action.

[Study on the relationship between the polymorphism of p53 gene intron 7 and non-small cell lung cancer (NSCLC) and p53 mutation in NSCLC tissues].

OBJECTIVE: To investigate the relationship between p53 gene intron 7 polymorphism and non-small cell lung cancer (NSCLC). METHODS: One hundred and five patients with NSCLC and 100 controls were selected with case-control analysis. Polymerase chain reaction (PCR), Apa I restriction enzyme digestion and agarose gel electrophoretic separation were used to identify genotypes of p53 intron 7 in peripheral blood. Then, NSCLC biopsy tissues (n=64) and NSCLC paraffin-embedded tissues (n=40) were selected for mutation analysis. PCR products of p53 exons 5-8 were sequenced on an automated sequencer following the identification of intron 7 genotypes as previously described. RESULTS: In NSCLC patients, the homozygote positive for ApaI site in p53 intron 7 was 23.8%, the homozygote negative was 12.34%, and the heterozygote was 63.8%. Whereas in control group, the homozygote positive, the homozygote negative and the heterozygote were 44.0%, 11.0% and 45.0%, respectively (P<0.01). In the second part, mutation rate of p53 exons 5-8 was 20.0%, 50.0% and 52.9% in samples with ApaI positive, negative and heterozygotes, respectively (P<0.05). CONCLUSION: p53 intron 7 ApaI polymorphism may be associated with human NSCLC.

[Medical thoracoscopy in the diagnosis of complicate pleural effusion].

OBJECTIVE: To evaluate the role of medical thoracoscopy in the diagnosis of the pleural effusion of unknown etiology. METHODS: The results of 36 patients with the pleural disease of unknown etiology diagnosed by medical thoracoscopy were retrospectively analyzed, including the pathologic results and the complications. RESULTS: Among the 36 patients, 35 were determined with positive rate of 97.2%, and no serious complications was found. CONCLUSION: Medical thoracoscopy is an important method of diagnosing complicate pleural effusion, and has high positive rate. It is a simple operation, with no serious complication, and fast recovery.

[Study on apoptosis of human lung adenocarcinoma cell line A549/DDP induced by ginsenoside Rh2 in vitro].

BACKGROUND: Lung cancer is one of the leading causes of cancer-related death in mankind. To exploit antitumor drug from plant has been a highlight at home and abroad. The aim of this study is to investigate the apoptosis of human lung adenocarcinoma cell line A549/DDP induced by ginsenoside Rh2 (G-Rh2) and to explore its possible molecular mechanism. METHODS: The growth inhibition effect of G-Rh2 on A549/DDP cells was evaluated by MTT assay. Cell cycle analysis, apoptosis index and tumor related gene expression were detected by flow cytometry. The changes of sApo-1/Fas level in the cell culture supernatant were determined by ELISA method. RESULTS: (1) G-Rh2 significantly inhibited the growth of A549/DDP cells in a dose-time-de-pendent manner. (2) After 24 hours' treatment with G-Rh2, apoptosis index of trial group was significantly higher than that of control group (P < 0.001). The proportion of cells in G0/G1 phase in trial group was much higher than that in control group (P < 0.01), while proportion in S phase in trial group was markedly lower than that in control group (P < 0.01). There was no significant difference in proportion in G2/M phase between trial group and control group (P > 0.05). (3) The positive expression rate of p53 and Fas in trial group was significantly higher than that in control group (P < 0.01, P < 0.001), while the positive expression rate of Bcl-2 in trial group was significantly lower than that in control group (P < 0.001). (4) The level of sApo-1/Fas in A549/DDP cell culture supernatant in trial group was remarkably lower than that in control group (P < 0.05). CONCLUSIONS: G-Rh2 can induce the apoptosis of A549/DDP cells. Its molecular mechanism may be up-regulating expression of p53 and Fas and down-regulating expression of Bcl-2.

[Effects of recombinant human tumor-necrosis factor related to apoptosis inducing ligand protein on apoptosis of human lung adenocarcinoma cell line resistant to cisplatin].

BACKGROUND: Tumor-necrosis factor related to apoptosis inducing ligand protein(TRAIL), like tumor-necrosis factor (TNF) and Fas, is a member of TNF cytokine supper family. Many researches have showed that TNF-α can reverse the resistance to some chemotherapeutic agents in cancer cell lines, and some anticancer drugs can result in up-regulations of death receptor (DR) and further lead to the enhancement of apoptosis induced by TRAIL. In order to clarify if TRAIL can reverse the resistance to cisplatin in cancer cells, the effects of recombinant human tumor-necrosis factor related to apoptosis inducing ligand protein (rhTRAIL) on apoptosis in human lung adenocarcinoma cell lines resistant to cisplatin (DDP) in vitro was explored. METHODS: Human lung adenocarcinoma cell lines resistant to cisplatin, A549/DDP cells, were cultured in regular condition. At 24 hours after TRAIL and DDP, alone or combined, microculture tetrazolium (MTT) dye was used to evaluate the cytotoxic effects. And besides, to detect the apoptotic effects of rhTRAIL on A549/DDP cells, flow cytometry assay was used to test the apoptosis proportion, diphenylamine assay (DPA) was applied to detect the percent of DNA fragmentation and Caspase-3 chluorometric assay was performed to test the activity of Caspase-3 among these cells. RESULTS: A549/DDP cells were not sensitive to low-dose rhTRAIL alone. The rate of growth inhibition and the apoptotic indexes such as the apoptosis proportion, the percent of DNA fragmentation and the activity of Caspase-3, had all no significant changes with rhTRAIL concentration less than 25µg/L (P > 0.05). But treated with higher-dose rhTRAIL more than 50µg/L, the four values changed obviously: 68.6%, (27.13± 0.66)%, (37.4±2.0)% and 0.117±0.011, respectively (P < 0.05). With combination of different concentration of rhTRAIL and 3mg/L DDP, the cyto-toxic and apoptotic effect was comparatively more apparent. The combination of rhTRAIL and 3mg/L DDP presented synergistic effect on A549/DDP, 12.5µg/L concentration of rhTRAIL together with 3mg/L DDP could kill 30.4% of A549/DDP cells. Furthermore, the rate of cell apoptosis, percent of DNA fragmentation and activity of caspase-3 increased to (19.39±0.54)%,(17.3±4.1)% and 0.138±0.009, which were significantly different from those of rhTRAIL alone (P < 0.01). CONCLUSIONS: High-dose rhTRAIL can also induce the cells resistant to cisplatin to apoptosis, but the cytotoxic and apoptotic effects of rhTRAIL alone are weaker than those of combination of rhTRAIL and low-dose cisplatin which can augment the apoptotic effect induced by rhTRAIL. rhTRAIL is expected to be an efficient biologic drug for treatment of lung cancer resistant to chemotherapy.

[Expression of COX-2 and its prognostic significance in non-small cell lung cancer].

BACKGROUND: To investigate the expression of COX-2 and its relation to clinical pathophysiological features and prognosis in non-small cell lung cancer (NSCLC). METHODS: The expression of COX-2 protein was detected in 52 NSCLC tissues by immunohistochemical (S-P) method. RESULTS: The positive COX-2 expression was observed in 25 (48.1%) cases of NSCLC tissues. The positive rate of COX-2 expression was 76.5% and 34.3% in adenocarcinoma and squamous cell carcinoma respectively (P < 0.01). The positive rate of COX-2 expression in T3+T4 disease (92.3%) was remarkably higher than that in stage T1+T2 (33.3%) (P < 0.01). There was a remarkable difference in COX-2 expression rate between clinical stage I+II (28.1%) and clinical stage III+IV (80.0%) groups (P < 0.01). The positive rate of COX-2 expression was 83.3% in those with lymph node metastasis, but only 17.9% in those without lymph node metastasis (P < 0.01). In addition, there were significant differences in positive rate of COX-2 expression among patients with ≤2, > 2 but < 5, ≥5 years of survival span respectively (P < 0.01). CONCLUSIONS: Overexpression of COX-2 in NSCLC, especially in adenocarcinoma, is closely related to invasion, lymph node metastasis and clinical stage of lung cancer. It may play a role in development of NSCLC, and also may be a prognostic marker.

[Clinical value of tumor supplied group of factor combined with CEA in diagnosing tuberculosis pleural effusion and malignant pleural effusion].

OBJECTIVES: To determine the clinical value of tumor supplied group of factor (TSGF) combined with carcinoembryonic antigen (CEA) in diagnosing tukerculosis pleural effusion and malignant pleurul effusion. METHODS: TSGF and CEA were detected by ELISA in 14 patients with tuberculosis pleural effusion and 28 patients with malignant pleural effusion. RESULTS: The average levels of TSGF and CEA in patients with malignant pleural effusion were higher than those with tuberculosis pleural effusion. The diagnostic sensitivity of TSGF and CEA in the malignant pleural effusion was 67.5% and 57.5%, the specificity was 85.7% and 78.6%, and the agreement rate was 72.2% and 62.9% respectively; while the sensitivity, specificity and agreement rate of TSGF combined with CEA were 93%, 67.4%, and 88.9%. CONCLUSION: The sensitivity and agreement rate of TSGF combined with CEA are higher than those of either of the two methods. TSGF combined with CEA is important in differentiating tuberculosis and malignant pleural effusion.