[Consensus on collaborative ethical review of multi-center clinical trials of new drugs of traditional Chinese medicine (version 1.0)].

At present, the issues regarding multi-center clinical trials of new drugs of traditional Chinese medicine(TCM) remain: the lack of agreement on the content and scope of the ethical review among the ethics committee members of the center and the participating units results in repeated review, which leads to a time-consuming ethical review process. Moreover, the review capabilities of the ethics committees of various research centers are uneven, which is not necessarily beneficial to the protection of subjects' rights and safety. In view of the existing problems, to improve the efficiency of ethical review of multi-center clinical trials of new drugs of TCM and avoid repeated reviews, the TCM Clinical Evaluation Professional Committee of Chinese Pharmaceutical Association organized experts to formulate the "Consensus on collaborative ethical review of multi-center clinical trials of new drugs of TCM(version 1.0)"(hereinafter referred to as "Consensus"). The "Consensus" is formulated in accordance with the requirements of relevant documents such as but not limited to "the opinions on deepening the reform of the evaluation and approval system to encourage the innovation of pharmaceutical medical devices", "the regulations of ethical review of biomedical research involving human subjects". The "Consensus" covers the scope of application, formulation principles, conditions for the ethics committee of the center, sharing of ethical review resources, scope and procedure of collaborative review, rights and obligations, etc. The aims of the "Consensus" is to preliminarily explore and establish a scientific and operable ethical review procedure. Additionally, on the basis of fully protecting the rights and interests of the subjects, a collaborative ethical review agreement needs to be signed to clarify the ethical review responsibilities of all parties, to avoid repeated review, and to improve the efficiency and quality of ethical review in multi-center clinical trials of new drugs of TCM.

miR-628-5p promotes growth and migration of osteosarcoma by targeting IFI44L.

This study investigated the role of miR-628-5p and interferon-induced protein 44-like (IFI44L) in osteosarcoma (OS) and determined whether miR-628-5p modulated OS growth by regulating IFI44L. Based on the data downloaded from Gene Expression Omnibus (GEO) database, we revealed that the expression of IFI44L was downregulated in OS and low expression of IFI44L was correlated with better prognosis of patients with OS. Biological prediction of its upstream regulatory miRNAs on the miRWalk website found that miR-628-5p is a possible upstream regulatory miRNA of IFI44L. Luciferase activity assay demonstrated that miR-628-5p could bind to the 3' untranslated region (UTR) of IFI44L, which proved the above prediction. The expression of miR-628-5p is upregulated in OS and high expression of miR-628-5p is correlated with poor prognosis of patients with OS. The results of RT-qPCR showed that the expression of miR-628-5p in MG-63, U2OS, Saos-2, and SW1353 cells was significantly higher than that in the hFOB1.19 cells. Downregulation of miR-628-5p by miR-628-5p inhibitor significantly inhibited the proliferation, migration, and invasion of MG-63 cells. By rescue assay, we found that knockdown of IFI44L rescued the proliferation and motility of miR-628-5p depleted MG-63 cells. Collectively, our present data illustrated that miR-628-5p promoted the growth and motility of OS at least partly by targeting IFI44L. Moreover, miR-628-5p and IFI44L might be proposed as promising biomarkers in OS diagnosis and treatment.

Significance of NKX2-1 as a biomarker for clinical prognosis, immune infiltration, and drug therapy in lung squamous cell carcinoma.

Background: This study was performed to determine the biological processes in which NKX2-1 is involved and thus its role in the development of lung squamous cell carcinoma (LUSC) toward improving the prognosis and treatment of LUSC. Methods: Raw RNA sequencing (RNA-seq) data of LUSC from The Cancer Genome Atlas (TCGA) were used in bioinformatics analysis to characterize NKX2-1 expression levels in tumor and normal tissues. Survival analysis of Kaplan-Meier curve, the time-dependent receiver operating characteristic (ROC) curve, and a nomogram were used to analyze the prognosis value of NKX2-1 for LUSC in terms of overall survival (OS) and progression-free survival (PFS). Then, differentially expressed genes (DEGs) were identified, and Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Gene Set Enrichment Analysis (GSEA) were used to clarify the biological mechanisms potentially involved in the development of LUSC. Moreover, the correlation between the NKX2-1 expression level and tumor mutation burden (TMB), tumor microenvironment (TME), and immune cell infiltration revealed that NKX2-1 participates in the development of LUSC. Finally, we studied the effects of NKX2-1 on drug therapy. To validate the protein and gene expression levels of NKX2-1 in LUSC, we employed immunohistochemistry(IHC) datasets, The Gene Expression Omnibus (GEO) database, and qRT-PCR analysis. Results: NKX2-1 expression levels were significantly lower in LUSC than in normal lung tissue. It significantly differed in gender, stage and N classification. The survival analysis revealed that high expression of NKX2-1 had shorter OS and PFS in LUSC. The multivariate Cox regression hazard model showed the NKX2-1 expression as an independent prognostic factor. Then, the nomogram predicted LUSC prognosis. There are 51 upregulated DEGs and 49 downregulated DEGs in the NKX2-1 high-level groups. GO, KEGG and GSEA analysis revealed that DEGs were enriched in cell cycle and DNA replication.The TME results show that NKX2-1 expression was positively associated with mast cells resting, neutrophils, monocytes, T cells CD4 memory resting, and M2 macrophages but negatively associated with M1 macrophages. The TMB correlated negatively with NKX2-1 expression. The pharmacotherapy had great sensitivity in the NKX2-1 low-level group, the immunotherapy is no significant difference in the NKX2-1 low-level and high-level groups. The analysis of GEO data demonstrated concurrence with TCGA results. IHC revealed NKX2-1 protein expression in tumor tissues of both LUAD and LUSC. Meanwhile qRT-PCR analysis indicated a significantly lower NKX2-1 expression level in LUSC compared to LUAD. These qRT-PCR findings were consistent with co-expression analysis of NKX2-1. Conclusion: We conclude that NKX2-1 is a potential biomarker for prognosis and treatment LUSC. A new insights of NKX2-1 in LUSC is still needed further research.

Segment selection for fusion and artificial disc replacement in the hybrid surgical treatment of noncontiguous cervical spondylosis: a finite element analysis.

Introduction: The treatment of skip-level cervical degenerative disease (CDD) with no degenerative changes observed in the intervening segment (IS) is complicated. This research aims to provide a reference basis for selecting treatment approaches for noncontiguous CDD. Methods: To establish accurate finite element models (FEMs), this study included computed tomography (CT) data from 21 patients with CDD (10 males and 11 females) for modeling. The study primarily discusses four cross-segment surgical approaches: upper (C3/4) anterior cervical discectomy and fusion (ACDF) and lower (C5/6) cervical disc arthroplasty (CDA), FA model; upper CDA (C3/4) and lower ACDF (C5/6), AF model; upper ACDF (C3/4) and lower ACDF (C5/6), FF model; upper CDA (C3/4) and lower CDA (C5/6), AA model. An initial axial load of 73.6 N was applied at the motion center using the follower load technique. A moment of 1.0 Nm was applied at the center of the C2 vertebra to simulate the overall motion of the model. The statistical analysis was conducted using STATA version 14.0. Statistical significance was defined as a p value less than 0.05. Results: The AA group had significantly greater ROM in flexion and axial rotation in other segments compared to the FA group (p < 0.05). The FA group consistently exhibited higher average intervertebral disc pressure in C2/3 during all motions compared to the AF group (p < 0.001); however, the FA group displayed lower average intervertebral disc pressure in C6/7 during all motions (p < 0.05). The AA group had lower facet joint contact stresses during extension in all segments compared to the AF group (p < 0.05). The FA group exhibited significantly higher facet joint contact stresses during extension in C2/3 (p < 0.001) and C6/7 (p < 0.001) compared to the AF group. Discussion: The use of skip-level CDA is recommended for the treatment of non-contiguous CDD. The FA construct shows superior biomechanical performance compared to the AF construct.

Association between Chinese herbal medicine (CHM) treatment and depression among cancer patients in China: An outpatient-based cross-sectional study.

Depression is a prevalent condition among cancer patients and significantly impacts their quality of life. Traditional Chinese Medicine, particularly Chinese Herbal Medicine (CHM), has shown potential in both anti-tumor and anti-depressive effects. However, there is a dearth of scientific literature exploring the association between CHM treatment and depression in cancer patients. This study aims to investigate the relationship between CHM treatment and depression in cancer patients. A cross-sectional study was conducted among cancer outpatients at Longhua Hosiptal, Shanghai University of Traditional Chinese Medicine, from June 2020 to April 2021 (Ethical approval number: 2020LCSY057). All patients signed informed consent and completed The European Organization for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire (EORTC QLQ-C30). Hamilton depression scale was evaluated depression by psychiatrists. The power of the sample size was determined using Gpower statistical and SPSS were used for statistical analysis. A total of 809 completed the study. Gender, medical insurance, the classification of time since diagnosis, ECOG, cancer stage, metastasis, gene mutation, treatment plan and CHM treatment were an important factor affecting depression (P < .05). Further analysis investigated the impact of CHM treatment on depression. There were 374 enrolled in CHM group and 435 enrolled in Non-CHM group. The assessment results of Hamilton depression scale and EORTC QLQ-C30 in CHM group were superior to those in Non-CHM group. The morbidity of depression is 50.27% in CHM group and 66.44% in Non-CHM group. After adjusting for potential confounders (gender, medical insurance, cancer stage, etc.), CHM treatment indicated negative correlation with depression (Odds ratio (OR) = 0.7, 95% confidence interval (CI): 0.5-0.9, P = .020). The interaction effects within each subgroup were no significantly affect the relationship between CHM treatment and depression (P > .05). CHM treatment was an independent protective factor for depression in cancer patients, and lead to better quality of life for cancer patients.

Biomechanical effects of hybrid constructions in the treatment of noncontinuous cervical spondylopathy: a finite element analysis.

BACKGROUND: Hybrid construction (HC) may be an ideal surgical strategy than noncontinuous total disc replacement (TDR) and noncontinuous anterior cervical discectomy and fusion (ACDF) in the treatment of noncontinuous cervical spondylopathy. However, there is still no consensus on the segmental selection for ACDF or TDR in HC. The study aims to analyse the effects of different segment selection of TDR and ACDF on cervical biomechanical characteristics after HC surgery. METHODS: Twelve FEMs of C2-C7 were constructed based on CT images of 12 mild cervical spondylopathy volunteers. Two kinds of HC were introduced in our study: Fusion-arthroplasty group (Group 1), upper-level (C3/4) ACDF, and lower-level TDR (C5/6); Arthroplasty-fusion group (Group 2), upper-level (C3/4) TDR and lower-level ACDF (C5/6). The follow-load technique was simulated by applying an axial initial load of 73.6 N through the motion centre of FEM. A bending moment of 1.0 Nm was applied to the centre of C2 in all FEMs. Statistical analysis was carried out by SPSS 26.0. The significance threshold was 5% (P < 0.05). RESULTS: In the comparison of ROMs between Group 1 and Group 2, the ROM in extension (P = 0.016), and lateral bending (P = 0.038) of C4/5 were significantly higher in Group 1 group. The average intervertebral disc pressures at C2/3 in all directions were significantly higher in Group 1 than those in Group 2 (P < 0.005). The average contact forces in facet joints of C2/3 (P = 0.007) were significantly more than that in Group 2; however, the average contact forces in facet joints of C6/7 (P < 0.001) in Group 1 group were significantly less than that in Group 2. CONCLUSIONS: Arthroplasty-fusion is preferred for intervertebral disc degeneration in adjacent upper segments. Fusion-arthroplasty is preferred for patients with lower intervertebral disc degeneration or lower posterior column degeneration. TRIAL REGISTRATION: This research was registered in Chinese Clinical Trial Registry (ChiCTR1900020513).

[Effects of Chinese herbal medicine Feiyanning Decoction on the ratio of CD4+CD25+ regulatory T cells and expression of transcription factor Foxp3 in mice bearing Lewis lung carcinoma].

OBJECTIVE: To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on the ratio of CD4(+)CD25(+) regulatory T cells and expression of transcription factor Foxp3 in mice with Lewis lung cancer. METHODS: Thirty-two male wild-type C57BL/6 mice aging from 6 to 8 weeks were inoculated with Lewis lung cancer cells to establish the tumor-bearing model of Lewis lung carcinoma and were randomly divided into model group, Chinese medicine group, chemotherapy group and Chinese medicine combined with chemotherapy group. After intervention for 14 d with corresponding drugs, behaviors, physical signs and changes of feed consumption of the mice were observed. All mice were sacrificed after drug treatment, and tumors and organs were removed to weigh and calculate organ indexes (lung index, spleen index and thymus index). The percentages of CD4(+)CD25(+) regulatory T cells in the thymus, spleen and tumor were determined by flow cytometry. The expression of Foxp3 mRNA in the thymus, spleen and tumor tissues was detected by quantitative real-time polymerase chain reaction. RESULTS: Compared with those in the model group, the mice in the Chinese medicine group showed significant reductions in spleen and thymus indexes and tumor weight, and elevation in the body weight without tumor (P<0.05). The numbers of CD4(+)CD25(+) regulatory T cells in spleen, thymus and tumor were lower in the Chinese medicine group than in the model group (P<0.05). The expression of Foxp3 mRNA in spleen, thymus and tumor was significantly down-regulated in the Chinese medicine group compared with the model group (P<0.05). There were no significant differences in CD4(+)CD25(+) regulatory T cell ratio and Foxp3 mRNA expression between the Chinese medicine combined with chemotherapy group and the chemotherapy group. CONCLUSION: Feiyanning Decoction can enhance the antitumor immune response and thus play a role in antitumor therapy by reducing the ratio of CD4(+)CD25(+) regulatory T cells and down-regulating the expression of Foxp3 mRNA.

Effects of Feiyanning Decoction, a compound traditional Chinese medicine, on iNOS and COX-2 expressions induced by tumor necrosis factor-α in lung adenocarcinoma cell line.

OBJECTIVE: To study the effect of Feiyanning Decoction (FYN), a compound traditional Chinese medicine, on expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by tumor necrosis factor-α (TNF-α) in human lung adenocarcinoma epithelial cell line (A549). METHODS: A549 cells were incubated with rat serum containing FYN for 24 h. Gene expressions of iNOS and COX-2 were determined by quantitative real-time polymerase chain reaction and Western blot. The iNOS-dependent luciferase reporter was transfected for 24 h and the cells were treated with the reagents for 24 h, then the transcriptional activity of iNOS promoter was detected by luciferase assay. The production of NO was determined by diaminofluorescein-2. RESULTS: FYN significantly inhibited TNF-α-induced expression of iNOS and COX-2 compared with the control group in A549 cells (P<0.01, P<0.01). Also, FYN inhibited the transcriptional activity of the iNOS promoter and reduced NO production compared with the control group (P<0.01, P<0.01). CONCLUSION: These results suggest that FYN inhibits iNOS and COX-2 activation induced by TNF-α, therefore, it is expected to develop a new strategy to treat lung cancer.

[Effects of Chinese herbal medicine Feiyanning decoction on expressions of nucleosome conformation-regulating factors H3-K56, Rtt109, Asf1 and E2F1 in Lewis-bearing mice].

OBJECTIVE: Malignant tumor cells were found with an abnormal cell cycle. Previous in vivo experiment had confirmed the Feiyanning's intervention effect on checkpoint signaling of G1/S in the cell cycle. This study was to further observe the expressions of nucleosome conformation-regulating factors intervened by Feiyanning decoction in S phase. METHODS: Lewis lung carcinoma models of C57BL/6 mice were established. Sixty mice were randomly divided into four groups: normal control group, model control group, Feiyanning group, and cisplatin group. There were 15 mice in each group. Tumor weight and tumor inhibition rate were observed. In addition, the cell cycle distribution was detected by flow cytometry and the proliferation index was calculated. Furthermore, mRNA and protein expressions of H3-K56, regulator of Ty1 transposition 109 (Rtt109), antisilencing function 1 (Asf1) and E2F1 were analyzed by real-time polymerase chain reaction and Western blot methods, respectively. RESULTS: The tumor weights of mice in the Feiyanning group and the cisplatin group were lower than those in the model group (P<0.01), with tumor inhibition rates of 27.92% and 42.50%, respectively. Cancer cell proliferation index and proportion of cancer cell population in S phase in the Feiyanning group were significantly lower than those in the cisplatin group (P<0.01). The mRNA and protein levels of H3-K56, Rtt109, Asf1, E2F1 in the Feiyanning group were lower than those in the model group (P<0.05). CONCLUSION: Feiyanning plays a role in intervening in the abnormal cell cycle by nucleosome conformation-regulating factors and thus inhibits the lung cancer cell proliferation.

[Traditional Chinese medicine for cancer pain].

Pain is one of the common symptoms of cancer which seriously affects the quality of life of the patients. Cancer pain is mainly treated with the three-step method, biological therapy or nerve block therapy based on antitumor therapy. However, up to 50 percent of patients with cancer-related pain do not receive adequate pain relief, affecting their physical and psychological well-being, and leading to a lower quality of life for the patient after conventional treatment. Clinical observation suggests that traditional Chinese medicine may alleviate cancer-related pain either by oral administration, topical administration, acupuncture or other means with continuing non-addictive and non-drug-resistant qualities. However, scientific evaluation of the efficacy of herbs in the treatment of pain is insufficient; the underlying mechanisms are unclear and, safety and toxicity remain a concern.

Development of shipboard automatic flow injection analysis-Surface-enhanced Raman spectroscopy instrument toward onsite detection of trace polycyclic aromatic hydrocarbons in water environment.

The qualitative and quantitative analysis of polycyclic aromatic hydrocarbons (PAHs) has been important for the environmental control of persistent organic pollutants for decades. Considering the potential risk of deterioration, degradation, and external pollution during transportation, the development of rapid and onsite detection of trace PAHs is in demand. Here, taking the advantage of high sensitivity of surface-enhanced Raman spectroscopy (SERS), we developed a shipboard instrument by combining a portable Raman instrument and a flow injection device, integrating the sample pretreatment and target detection step by step. The feasibility of the instrument was demonstrated by detecting trace benzo[a]pyrene from different water environments with the lowest detection concentration less than 1 µg/l. The reliable stability and repeatability indicate that in the case of emergency response, the developed flow injection analysis-SERS instrument is very promising for the quantitative and qualitative analysis of diverse organic pollutants other than PAHs in water environments.

Trace detection of polycyclic aromatic hydrocarbons in environmental waters by SERS.

Polycyclic aromatic hydrocarbons (PAHs) are among the most hazardous pollutants and have attracted significant attention in the last decades. Up to now, rapid and on-site trace detection of PAHs remains a challenging issue. Here, taking advantage of the high sensitivity and reliable qualification of Surface-enhanced Raman Spectroscopy (SERS), we firstly carried out trace analyses of 16 typical PAHs in water at concentrations as low as 100-0.1 µg/L, depending on the number of aromatic rings of the molecule. Furthermore, owing to the simplicity of the liquid-liquid extraction (LLE) step, the sensitivity was further improved 2-3 orders of magnitude, and the lowest detectable concentrations were 100, 50, and 5 ng/L for anthracene, pyrene, and benzo[a]pyrene (the three PAHs typically found in heavily polluted waters), respectively. The LLE-SERS approach was successfully applied to the qualitative and quantitative analyses of different (ocean and coast) water samples being spiked by these three PAHs, which showed great promise as a trace detection tool of PAHs under water environments having different contaminant matrices.

Involvement of caspase-3 activity and survivin downregulation in cinobufocini-induced apoptosis in A 549 cells.

Cinobufocini injection is a preparation containing water-soluble components of the toad skin. The aim of the present study was to investigate the apoptosis of human lung adenocarcinoma cell line A 549 induced by cinobufocini. A 549 or HLF-1(human lung fibroblast) cells were treated with cinobufocini at different concentrations for 24 and 48 h, respectively. The proliferation of cells was detected with the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Morphology of cells was carried out with scanning electronic microscopy (SEM) and Hoechst 33258 staining. The apoptosis rate was examined by flow cytometry. The expression of survivin was examined with RT-PCR and Western blot assay. The caspase-3 and caspase-7 activities were detected with caspase colorimetric protease assay. We found that cinobufocini significantly inhibited tumor growth of A 549 cells in a dose- and time-dependent manner without damaging non-cancerous cells (HLF-1) and induced granular apoptotic bodies of A 549 cells. Next, cinobufocini increased the percentage of cells in G1 phase and decreased the percentage of cells in S phase in A 549 cells. Furthermore, cinobufocini downregulated the expression of survivin mRNA and protein. Finally, cinobufocini upregulated caspase-3 activity. We concluded that cinobufocini induces apoptosis of A 549 cells, which is associated with the decreasing expression of survivin mRNA and protein, increasing caspase-3 activity of A 549 cells.

[Effect of felyanning granule in antagonizing Lewis lung cancer cell proliferation through cell cycle G1/S checkpoint dominating signaling intervention].

OBJECTIVE: To observe the effect of Feiyanning Granule (FYN) on tumor growth and cell cycle distribution in mice with Lewis lung cancer, as well as its influence on G1/S cell cycle checkpoint dominating signaling RB-E2F1 bio-axis. METHODS: Modeled C57BL/6 mice were randomly divided into 6 groups: the model group (A), the DDP treated group (B) peritoneally injected with cisplatin 0.1 mg on d1, d3 and d5 after modeling, and the 4 FYN treated groups (C-F), administered via gastrogavage with FYN Decoction, and FYN Granule in small-, median- and high- dose respectively for 14 days. The tumour inhibiting rate, tumour weight, and body weight of mice were observed after treatment; cell cycle distribution was detected by flow cytometry (FCM), RB-E2F1mRNA expressions in tumour tissue were analyzed by RT-PCR, and their protein expressions by Western blot. RESULTS: Tumour weight in the 5 treated groups was lower than that in the model group (P < 0.05, P < 0.01). Body weight in group E was significantly higher than that in group A and B (P < 0.05, P < 0.01). FCM analysis showed the proportion of G0/G1 phase was higher in group E than in group A, B and C (P < 0.01), and cancer cell proliferation index (PI) in group E was lower than in group B (P < 0.05, P < 0.01). RT-PCR showed mRNA level of E2F1 was lower, but that of RB was significantly higher in group E than those in group A, B and C respectively (P < 0.01). Westem blot analysis showed the protein expression of E2F1 was lower in group E and B than that in group A (P < 0.05), while the protein expression of Rb in group E was higher than that in group A, B and C (P < 0.05). CONCLUSION: The effect of FYN in inhibiting Lewis lung cancer growth was related to its intervention on cancer cell cycle distribution which blocks most tumor cells in G0/G1 phase. Moreover, FYN can reduce MDM2 expression, enhance P53 expression to influence cell cycle G1/S checkpoint dominating signaling, so as to achieve the effect of antagonizing lung cancer cell proliferation.

Effects of Feiyanning Decoction on gene expression of nuclear factor-kappaB activated by tumor necrosis factor-alpha in lung adenocarcinoma cell line.

OBJECTIVE: To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on gene expression of nuclear factor-kappaB (NF-kappaB) activated by tumor necrosis factor-alpha (TNF-alpha) in lung adenocarcinoma cell line (A549). METHODS: A549 cells were incubated with rat serum containing Feiyanning at different concentrations for 24 and 48 h, respectively. Morphology of cells was observed by an inverted microscope after treatment with reagents. The cell proliferation was examined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) assay. The expressions of NF-kappaB and inhibitor kappaBalpha (IkappaBalpha) were studied by Western blotting. NF-kappaB-dependent luciferase reporter (3 x kappaB-luc) was transfected for 24 h, and the cells were treated with the reagents for 24 h, and then the transcriptional activity of NF-kappaB promoter was detected by luciferase assay. RESULTS: TNF-alpha (1 microg/L) strongly induced the expression of NF-kappaB by approximately 1.76-fold compared with the control in the nuclei of A549 cells, and the induced NF-kappaB expression was significantly suppressed by addition of Feiyanning (P < 0.01). In addition, Feiyanning inhibited the transcriptional activity of the NF-kappaB promoter. However, we observed no significant changes in IkappaBalpha expression (P > 0.05). CONCLUSION: Feiyanning Decoction can markedly inhibit human lung cancer A549 cell proliferation, which may be partly due to inhibition of NF-kappaB activation induced by TNF-alpha. It is therefore expected to be a new strategy for treating lung cancer.

Effects of Feiyanning Decoction on proliferation of lung adenocarcinoma cell line and their production of interleukin-6 and interleukin-8 induced by tumor necrosis factor-alpha.

OBJECTIVE: To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on proliferation of lung adenocarcinoma cell line A549 cells and their production of interleukin-6 (IL-6) and IL-8 induced by tumor necrosis factor-alpha (TNF-alpha). METHODS: A549 cells were incubated with rat serum containing Feiyanning Decoction at 15% for 24, 48 and 72 h respectively. The cell proliferation was examined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium, monosodium salt assay (WST-8). The production of IL-6 and IL-8 was tested by enzyme-linked immunosorbent assay after 48-hour treatment of reagents, and the expressions of IL-6 and IL-8 mRNAs were detected by reverse transcription-polymerase chain reaction. RESULTS: Serum containing Feiyanning Decoction had obvious inhibitive functions in A549 cell proliferation after 48- and 72-treatment. TNF-alpha (1 microg/L) strongly induced the production of IL-6 and IL-8 as compared with the control serum in A549 cells, and the induced cytokine production was significantly suppressed by 15% serum containing Feiyanning Decoction (P<0.01). In addition, serum containing Feiyanning Decoction could inhibit the mRNA expressions of IL-6 and IL-8 (P<0.01). CONCLUSION: Feiyanning Decoction can inhibit IL-6 and IL-8 production induced by TNF-alpha. It is therefore expected to be a new strategy for treating lung cancer.

Pitavastatin inactivates NF-kappaB and decreases IL-6 production through Rho kinase pathway in MCF-7 cells.

The aim of the present study was to provide new mechanistic insight into the effect of pitavastatin at low dose on NF-kappaB activated by TNF-alpha in the human breast cancer cell line (MCF-7). We found that treatment of MCF-7 with 1 microM pitavastatin inhibited the proliferation and suppressed the nuclear expression of NF-kappaB p65 induced by TNF-alpha with Western blotting. Furthermore, EMSA showed that pitavastatin significantly reduced the DNA binding activity of NF-kappaB induced by TNF-alpha. Subsequently, luciferase assay revealed that pitavastatin (1 microM) inhibited the transcriptional activity of the NF-kappaB promoter, which was clearly related to the HMG-CoA reductase activity because addition of mevalonic acid (MEV) could elevate the NF-kappaB activity. Moreover, the Rho kinase inhibitor Y27632 abolished the effect of pitavastatin on NF-kappaB activity. Finally, the addition of TNF-alpha significantly increased IL-6 protein production, which was suppressed by the addition of pitavastatin. These results suggest that pitavastatin at low dose (1 microM) inhibits NF-kappaB activation and decreases IL-6 production induced by TNF-alpha. It is dependent on Rho kinase pathway in human breast cancer cells.

Downregulation of survivin expression and elevation of caspase-3 activity involved in pitavastatin-induced HepG 2 cell apoptosis.

The aim of the present study was to research the apoptosis of human hepatocellular carcinoma cell line HepG 2 induced by pitavastatin. HepG 2 cells were treated with increasing doses of pitavastatin or with mevalonic acid for 48 h. The proliferation of cells was detected with WST-8. The morphology of the nucleus was observed under a microscope by Hoechst 33258 staining. The apoptosis peaks were examined by flow cytometry. The expression of survivin mRNA was examined with RT-PCR. The caspase-3 activity was detected with caspase-3 colorimetric protease assay. We found that growth inhibitory effects were observed for treatment with pitavastatin at 10-50 microM. Pitavastatin at 10 microM induced granular apoptotic bodies of HepG 2 cells. Furthermore, pitavastatin at 10 microM increased the appearance of sub-G1 population of HepG 2 cells. Finally, pitavastatin at 10 microM downregulated the expression of survivin mRNA and upregulated the caspase-3 activity, which was clearly related to the HMG-CoA reductase activity. These results suggest that pitavastatin at 10 microM induces apoptosis of HepG 2 cells, which is associated with the decreased expression of survivin mRNA and increased caspase-3 activity of HepG 2 cells.

[Effects of NK-104 on apoptosis and caspase-3 activity in hepatocellular carcinoma cells].

OBJECTIVE: To study the apoptosis of human hepatocellular carcinoma cell line HepG 2 induced by pitavastatin (NK-104) and to investigate the change of caspase-3 activity. METHODS: HepG 2 cells were treated with NK-104 for 48 hours and observed under a microscope by using Hoechst 33258 staining. The proliferation of the cells was detected with WST-8 method. The apoptosis peaks were examined by flow cytometry. The caspase-3 activity was detected with caspase-3 colorimetric protease assay. RESULTS: The treatment of HepG 2 with 10 micromol/L NK-104 could inhibit the proliferation, and induce HepG 2 apoptosis and up-regulate the caspase-3 activity. CONCLUSION: NK-104 at 10 micromol/L induces apoptosis of HepG 2 cells. It is dependent on caspase-3 pathway in human hepatocellular carcinoma cells.

Enhancement of tissue engineered bone formation by a low pressure system improving cell seeding and medium perfusion into a porous scaffold.

To obtain more extensive bone formation in composites of porous ceramics and bone marrow stromal cells (BMSCs), we hypothesized that a low-pressure system would serve to facilitate the perfusion of larger number of BMSCs into the porous scaffold, enhancing bone formation within the composites. After culturing BMSCs in osteogenic medium, porous blocks of beta-tricalcium phosphate (beta-TCP) were soaked in the cell suspension. Composites of the block and BMSCs were put immediately into a vacuum desiccator. Low pressure was applied to the low pressure group, while controls were left at atmospheric pressure. Composites were incubated in vitro or subcutaneously implanted into syngeneic rats, then analyzed biologically and histologically. In the in vitro group, cell suspension volume, cell seeding efficiency, alkaline phosphatase (ALP) activity, and DNA content in the beta-TCP blocks were significantly higher in low pressure group than in the controls. Scanning electron microscopy (SEM) demonstrated that a greater number of cells covered the central parts of the composites in the low pressure group. ALP activity in the composites was increased at 3 and 6 weeks after implantation into rats. Histomorphometric analysis revealed more uniform and extensive bone formation in the low pressure group than in the controls. The application of low pressure during the seeding of BMSCs in perfusing medium into a porous scaffold is useful for tissue-engineered bone formation.