Adult ; Bronchoalveolar Lavage ; methods ; Bronchoalveolar Lavage Fluid ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; therapy

OBEJECTIVE Gecko has been clinically used in China for many years. It has been proved that the gecko polypeptide mixture(GPM)extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci-noma(HCC)HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1)for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres-sion of apoptosis- related proteins and endoplasmic reticulum stress (ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS)generation.In this report, we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The GPM could induce ROS generation and up-regulate ERs-related proteins. CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.

Objective To explore the effect of advanced glycation end products (AGEs) on proliferation of bone marrow mesenchymal stem cells (MSCs) and related mechanism.Methods The bone marrow MSCs were isolated from male Sprague Dawley rats and cultured in vitro.The flow cytometer was used to identify the bone marrow MSCs by detecting positive labels (CD29 and CD90) and negative labels (CD34 and CD45).The advanced glycation end products-bovine serum albumin (AGE-BSA),one of the AGEs,was used in this study.The methyl thiazolyl tetrazolium (MTT) method was used to detect the effect of AGE-BSA on proliferation of the bone marrow MSCs.In MTT test,there were 3 groups:AGE-BSA group,BSA group,and control group.In AGE-BSA group,different doses of AGE-BSA (0,25,50,100 and 200 μg/ml) was used to stimulate the bone marrow MSCs for 6 h,12 h or 24 h.In BSA group,the 200 μg/ml BSA was used to stimulate the bone marrow MSCs for 6 h,12 h or 24 h.Gene chips detection was used to detect change of genes expression in bone marrow MSCs.Results The proliferation of the bone marrow MSCs could be inhibited by AGE-BSA,in a dose- and time-related manner.Compared with the BSA group,after being treated with 100 μg/ml AGE-BSA for 24 h or 200 μg/ml AGE-BSA for 12 h and 24 h,the proliferation of the bone marrow MSCs decreased obviously.The gene chips detection found that there were changes in expression of 17 genes in the bone marrow MSCs after being treated with AGE-BSA (200 μ.g/ml) for 12 h or 24 h,and the genes were same at the two time points.Among 17 genes,the expression of 12 genes increased,including four inflammatory factors (CCL3,CCL2,CCL4 and IL-1β),and 5 genes decreased.Conclusion AGE-BSA can inhibit the proliferation of bone marrow MSCs,which may be related to the onset of the diabetic osteoporosis.

OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture (GPM). METHODS RNA-seq technology was used to identify the differen?tially expressed genes of human hepatocellular carcinoma (HCC) HepG2 cells treated with or without GPM. The HepG2 cells were treated with different concentration of GPM (0, 0.1, 0.2, 0.3, 0.4 mg·mL-1) for 6 h, 12 h and 24 h, respectively. MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells. Western blot analysis was applied to observe the expression of apoptosis-related proteins in HepG2 cells. RESULTS The results showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner. We applied many analysis methods, including differen?tially expressed genes analysis, Gene Ontology (GO) enrichment analysis, KEGG pathway enrichment analysis, protein- protein interaction network analysis to screen out possible molecular mechanisms. ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM. GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The mechanism is closely related to ERs, which might be beneficial for clinical therapy of HCC. CONCLUSION GPM can inhibit cells proliferation and induce apoptosis in HepG2 cells. The gene expression profile of GPM in HepG2 cells was obtained. The present study revealed the potential anti-tumor mechanism of GPM.

OBJECTIVE To explore the role of gecko crude peptides (GCPs) in the proliferation, apoptosis, migration and lymphangiogenesis of human hepatocellular carcinoma cells (HepG2) and human lymphaticendothelial cells (HLECs) in vitro. METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the anti- proliferative effect of GCPs and siRNA-VEGF-C on HepG2 cells, Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis. The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay. The protein and mRNA expressions of vascular endo?thelial growth factor-C (VEGF-C) and CXC chemokine receptor-4 (CXCR4) were detected by q-PCR, immunofluorescence, Western blot. The protein expressions of the extracellular signal regulated kinase (ERKI/2), c-Jun N-terminal kinase (JNK), p38-mitogen activated protein kinases (p38 MAPK), serine/threonine kinase (Akt) and phosphatidylinositol- 3- kinase (PI3K) were detected by western blot. The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay. The protein and mRNA expressions of vascular endothelial growth factor receptor-3 (VEGFR-3) and stromal cell-derived factor-1 (SDF-1) were detected by q-PCR, Western blot. RESULTS GCPs and siRNA-VEGF-C inhibited HepG2 proliferation, invasion and migration, and the most obvious inhibitory effect was both synergistic effects. Thus, GCPs suppressed HLECs proliferation, migration and tube-like structure formationin a dose- dependent manner, and had inhibitory effect of tumor- induced lymphangiogenesis in vitro. Additionally, we found that GCPs and siRNA- VEGF- C decreased the expressions of MMP-2, MMP-9, VEGF-C, CXCR4, phospho-ERK1/2, phospho-P38, phospho-JNK and PI3K in HepG2 cells. Moreover, GCPs had a dose-dependent depressive effecton the expressions of VEGFR- 3, SDF- 1 in HLECs. CONCLUSION The low expression of VEGF- C mediated by siRNA-VEGF-C and GCPs inhibit tumor proliferation, invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C, and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.

Seven meroterpenoids and five small-molecular precursors were isolated from Penicillium sp., an endophytic fungus from Dysosma versipellis. The structures of new compounds, 11beta-acetoxyisoaustinone (1) and isoberkedienolactone (2) were elucidated based on analysis of the spectral data, and the absolute configuration of 2 was established by TDDFT ECD calculation with satisfactory match to its experimental ECD data. Meroterpenoids originated tetraketide and pentaketide precursors, resepectively, were found to be simultaneously produced in specific fungus of Penicillium species. These compounds showed weak cytotoxicity in vitro against HCT-116, HepG2, BGC-823, NCI-H1650, and A2780 cell lines with IC 50 > 10 micromol x L(-1).
Berberidaceae ; microbiology ; Cell Line ; Cell Line, Tumor ; Humans ; Lactones ; isolation & purification ; pharmacology ; Monoterpenes ; isolation & purification ; pharmacology ; Penicillium ; chemistry

Objective To understand the mycoplasma pneumoniae(MP) infection degree in community children by testing specific IgM antibodies against MP in different age bracket group in Shanghai Meilong area. Methods Using random sampling method, blood specimens of 1 817 children from kindergartens and primary or junior high schools in Meilong area were obtained. Children were from 2 to 15 years old, 969 males, 848 females. The specimens were tested for IgM antibodies against MP using with gelatin particle agglutination test. The data were statistically analyzed using with ?2 test. Results Five hundred and fifty-nine (30.7%) IgM antibodies against MP were positive from 1 817 blood specimens. The positive percentages were 27.34% and 34.66% for males and females, which had significant difference(?2=11.383 P=0.001). The higher percentage was detected from kindergarten children than primary and junior high school children(P=0). The positive percentages of anti-mycoplasma IgM had no significant differences between different kindergartens and primary schools(P=0.526,0.232). On the contrary, between different junior high schools, there were siginificant differences (?2=9.825 P=0.002). Conclusions MP is an important pathogenic mycoplasma cause for respiratory tract infections in Meilong area. It is relative to childhood asthma. The prevention and cure of MP infection for children shall be paid more attention.

OBJECTIVETo study the metastasis feature of the primary and metastatic lymph node lesions in supraglottic or hypopharyngeal cancer.

METHODSThe expression of CD44 and nm23-H1 in specimens from the primary and metastatic lymph node lesions of the 41 cases with supraglottic or hypopharyngeal cancer were studied with immunohistochemistry method and flow cytometry.

RESULTSNo correlation was found between the expression of CD44, nm23-H1 and the tumor differentiation of the supraglottic or hypopharyngeal cancer, but their expression related with the clinical staging. The CD44 and nm23-H1 positive expression rates in the primary and metastatic lymph node lesions were 75.6% (31/41), 85.4% (35/41) and 34.1% (14/41), 26.8% (11/41) respectively (P >0.05). The average fluorescence index of CD44 and nm23-H1 in the primary and metastatic lymph node lesions were 1.27 +/- 0.18, 1.33 +/- 0.16 and 1.11 +/- 0.19, 1.08 +/- 0.15 (x +/- s) respectively (P >0.05).

CONCLUSIONSThe expressions of CD44 and nm23-H1 in the metastatic lymph node tumor had no difference compared with that in primary tumor of the supraglottic or hypopharyngeal cancer. The difference of metastasis potentials between the primary and metastatic lymph node lesions in the same patient was not proved in this study and should be further investigated from multiple oncogens markers.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Hyaluronan Receptors ; genetics ; Hypopharyngeal Neoplasms ; metabolism ; pathology ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; genetics ; Neoplasm Staging

Objective To investigate the possible molecular mechanisms of Gecko peptides mixture (GPM) and research to human hepatocellular carcinoma SMMC7721 cells on autophagy with GPM.Methods SMMC7721 cells were put into plates in its logarithmic phase,and they were treated with different concentration of GPM (0,0.04,0.06,0.09,0.14,0.20,0.30,0.45 mg · mL-1) for 24 h,and then detected corresponding indicators with respective methods.The viability of SMMC7721 cells was detected with 3-(4,5-dimethyl2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT).The concentration of normal group,GPM low-dose,middle-dose and high-dose experimental groups was separately 0,0.1,0.15,0.22 mg · mL-1,according to the results of MTT.Rapamycin was chosen as the positive control drug,which concentration was 40 μg · mL-1.The autophagy of different concentration GPM on SMMC7721 cells was detected by monodansylcadaverine (MDC) staining.Expression levels of Beclin1 and LC3 in SMMC7721 cells were measured by immunohistochemical method and Western blot assay.Results GPM could significantly inhibit the proliferation of SMMC7721 cells in a dose -dependent manner,and the IC50value was 0.16 mg · mL-1 for 24 h.MDC staining showed that there are plenty of autophagosome in cytoplasm,emerging bright green fluorescent in the fluorescence microscope after treatment with GPM for 6 h.The percentage number of positive cells of total cells number in normal group,control group,GPM low,middle and high-dose experimental groups were (15.70 ± 0.26)％,(63.47 ± 0.54)％,(17.09 ± 0.37)％,(70.66 ±0.56) ％,(78.48 ±0.68) ％,respectively.Compared with the normal group,control group,GPM middle and high-dose experimental groups increased,the differences were statistically significant (P ＜ 0.05).The indicators of LC3 in that five groups were (35.84 ± 0.36) ％,(82.41 ± 0.82) ％,(60.83 ± 0.61) ％,(69.66 ± 0.70) ％,(74.61 ± 0.75) ％,respectively.The differences between each group and normal group were statistically significant (P ＜ 0.05).The grayscale average ratio of Beclin1 and β actin in normal group,control group,GPM low,middle and high-dose experimental groups were (6.87 ± 0.68) ％,(11.26 ± 0.87) ％,(6.46 ± 1.34) ％,(11.58 ± 0.95) ％,(15.82 ± 1.58)％,respectively.Compared with the normal group,control group,GPM middle and high-dose experimental groups increased,the differences were statistically significant (P ＜ 0.05).The indicators of LC3 Ⅱ and βactin in that five groups were (26.70 ± 0.41) ％,(103.17 0.88) ％,(30.29 ± 0.52) ％,(36.32 ± 0.52) ％,(64.34 ± 0.48) ％.The differences between each group and normal group were statistically significant (P ＜ 0.05).Conclusion GPM could have an effect on the autophagy in human hepatocellular carcinoma SMMC7721 cells.The possible mechanism may be GPM induced autophagy of SMMC7721 cells,causing cell death ultimately.

OBJECTIVETo evaluate the clinical effects of Chinese medicine (CM) on acute myocardial infarction (AMI) with a prospective cohort study.

METHODSA total of 334 AMI patients from January 2007 to March 2009 were consecutively enrolled, and were assigned to a treatment group (169 cases) treated with combined therapy (CM for at least one month and Western medicine) and a control group (165 cases) with Western medicine alone. Clinical data including age, gender, smoking, medical history, infarction area, heart functional classification, CM syndrome scores, blood-stasis syndrome score, primary end-point (death, nonfatal myocardial infarction, and revascularization) and secondary end-point (ischemic stroke, rehospitalization due to angina, heart failure and shock), were collected. CM syndrome scores, blood-stasis syndrome score, primary end-point and secondary end-point were collected during the 6-month follow-up. Kaplan-Meier method was used for the survival analysis. The multifactor analysis was analyzed by Cox proportional hazards regression.

RESULTSAt the end of 6-month the CM syndrome score and bloodstasis syndrome score in the treatment group were lower than those in the control group (P<0.01), especially the symptoms of chest pain, spontaneous perspiration and insomnia. Rehospitalization rate due to angina during the 6-month follow-up in the treatment group (2.96%) was lower than that in the control group (7.88%, P<0.05). Kaplan- Meier survival curve showed that event-free cumulated survival of rehospitalization due to angina during the 6-month follow-up in the treatment group was higher than that in the control group (Log rank 4.700, P=0.03). Cox regression analysis showed that heart dysfunction [hazard ratio (HR)=1.601, 95% CI=1.084-2.364, P=0.018] and diabetes mellitus (HR=1.755, 95% CI=1.031-2.989, P=0.038) were hazard factors to end-point, whereas CM (HR 0.405, 95% CI=0.231-0.712, P=0.002), percutaneous coronary intervention (PCI, HR=0.352, 95% CI=0.204-0.607, P<0.001) and angiotensin converting enzyme (ACE) inhibitors (HR=0.541, 95% CI=0.313-0.936, P=0.028) were protective factors.

CONCLUSIONSCM therapy could decrease CM syndrome scores and blood-stasis syndrome score, reduce the rehospitalization rate during 6-month follow-up due to angina. Heart dysfunction and diabetes mellitus were hazard factors to end-point, whereas CM, PCI and ACE inhibitors were protective factors.

Adult ; Aged ; Case-Control Studies ; Cohort Studies ; Female ; Hematologic Diseases ; complications ; epidemiology ; Hospitalization ; statistics & numerical data ; Humans ; Male ; Medicine, Chinese Traditional ; adverse effects ; methods ; Middle Aged ; Myocardial Infarction ; complications ; epidemiology ; therapy ; Prospective Studies ; Research Design ; Syndrome ; Treatment Outcome