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ObjectiveTo study the effect of icariin on the proliferative capacity of hepatocellular carcinoma cell line CLC5 and the underlying mechanism. MethodThe targets of icariin were screened out by network pharmacology， and the target network and protein-protein interaction （PPI） network were constructed to predict the possible targets and pathways of icariin. CCK-8 assay was employed to explore the effects of different concentrations （0， 6.25， 12.5， 25， 50 μmol·L-1） of icariin on the viability of CLC5 cells. Further， CLC5 cells were treated with 0， 25， 50 μmol·L-1 icariin， and the effect of icariin on CLC5 cell proliferation was examined by Edu-488 assay and clone formation assay （CFA）. Western blot was employed to measure the expression levels of proteins in the protein kinase B （Akt）/glycogen synthase kinase 3β （GSK3β）/cell cycle-dependent kinase （CDK） pathway in the CLC5 cells exposed to different concentrations of icariin. ResultNetwork pharmacological analysis revealed that icariin may inhibit the hepatocellular carcinoma via cell cycle arrest and inhibition of tumor cell proliferation. Compared with the blank group， icariin decreased the viability of CLC5 cells in a time- and concentration-dependent manner （P<0.01） and reduced the positive rate of Edu-488 and the colonies in CFA （P<0.05， P<0.01）. Moreover， icariin down-regulated the protein levels of p-Akt， p-GSK3β， CDK4， and CyclinD1 （P<0.05， P<0.01）. ConclusionIcariin may block cell cycle to suppress the proliferation of CLC5 cells via inhibiting the Akt/GSK3β/CDK pathway.
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OBJECTIVE To study the effect of Cistanches phenylethanol glycosides (CPhGs) on acute lung injury in rats and the possible mechanism of action. METHODS Sixty SD rats were randomly divided into six groups: Normal control group, model group (lipopolysasccharide (LPS) 3 mg·kg-1], dexamethasone (Dex) positive control group (model+ Dex 2 mg·kg-1), model + CPhGs 125, 250 and 500 mg·kg-1 with 10 rats in each group. Each dose group of CPhGs was ig administered every day for 30 d, and the other three groups were ig administered with the same volume of distilled water for 30 d. The mode+Dex 2 mg·kg-1 group was ig given Dex 2 mg·kg-11 h before modeling. The rat model of acute lung injury was established after the rats in the other groups were intubated by the breath tube. Three hours after modeling, the rats were sacrificed by blood sampling from the abdominal aorta. The percentage of neutrophils in the whole blood, the activity of serum superoxide dismutase (SOD), glutathione (GSH) activity and the contents of malondialdehyde (MDA) and nitric oxide (NO) were detected. The contents of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β) and IL-6 in alveolar lavage fluid were detected. The wet/dry mass ratio (W/D) of the middle lobe of the right lung was calculated and the histopathological changes of the posterior lobe of the right lung were observed by HE staining. RESULTS Compared with normal control group, the percentage of neutrophils in the whole blood, W/D in the middle lobe of the right lung, the contents of MDA and NO in the serum were increased (P<0.05), but SOD, GSH, TNF-α and IL-6 were decreased in the model group (P<0.05). Compared with the model group, the percentage of neutrophils in the whole blood and W/D in right middle lobe of the right lung in the group of model+CPhGs 500 mg·kg-1 was decreased (P<0.05), but the contents of TNF-α and IL-6 in alveolar lavage fluid of rats decreased significantly, and histopathological observation showed that inflammation was alleviated to varying degrees. The activities of SOD and GSH in rat serum of the model + CPhGs 500 mg·kg-1 group were increased significantly (P<0.05), while the contents of MDA and NO were decreased significantly (P<0.05). CONCLUSION CPhGs have a protective effect against acute lung injury induced by LPS in rats, and its mechanism may be related to its antioxidant effect, alleviation of pulmonary edema and reduction of the release of inflammatory factors.
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To investigate the efficaciousness of breast-conserving therapy in connection with neoadjuvant chemotherapy on breast cancer. 68 patients, who were confirmed going down with breast cancer and hospitalized from June 2015 and June 2017, were sampled and divided into two groups using the random digit table, i.e. the observation group [n=34] and the control group [n=34]. Patients in the observation group experienced breast-conserving therapy integrated with neoadjuvant chemotherapy, but those in the control group received the radical resection of breast cancer. Patients' condition in surgery, incidence of post-surgery complications as well as patient survivals were compared and coded. In the observation group, surgical duration, intraoperative bleeding amount, length of stay in hospital and incidence rate of post-surgery complications were all lower than the patients with the similar conditions in the control group with evident distinctions in statistics [p<0.05]. In the observation group, survival ratios of one-to-five-year living patients were evidently higher than those in the control group. The distinctions owned evident significance in calculations [p<0.05]. In comparison of the recurrence ratio of disease and the rate of distant metastasis between the observation group [5.88% and 8.82%] and the control group [11.76% and 8.82%], differences had no statistical significance [p>0.05]. Before treatment, compared with the score of life quality in the two groups, no evident distinction in statistical exists [p>0.05], however, after that, the life quality in the observation group evidently outweighs the quality in the control group, which shows the distinctions in statistics [p<0.05]. Breast-conserving therapy in combination with neoadjuvant chemotherapy shows promising clinical value in ameliorating the life quality, decreasing the mortality rate and the incidence of adverse reaction, which is expected to be applied in clinical practices as a kind of safe and effective method
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The genetic structure of small-scale landscape groups of Oncomelania hupensis in Songzi City ,Hubei Province was identify in this study .O .hupensis snails were collected from 10 habitats in Songzi City ,of which 6 polymorphic microsat-ellite DNA loci (T6-17 ,P101 ,D11 ,B14 ,T4-33 ,and C22) were carried out with GeneScan .The number of alleles (Na) ,het-erozygosity (H) ,fixation index (FST) of snails in each group ,genetic distance between groups ,and the polymorphic informa-tion content (PIC) were calculated .Cluster analysis was then carried out based on genetic distance ,and hierarchical AMOVA calculation was conducted .By certified the shells of snails ,10 groups were divided into light and ribbed shell (including shallow rib and deep rib) .There were 141 alleles in total detection on 10 populations and 20-34 alleles in each locus ,which were detec-ted for 23 .5 on average .The average number of alleles in 6 loci was 1 .575 and the number of alleles in each locus was uneven , showing large numerical differences ranged from 0 .445 to 3 .060 .The average observed heterozygosity (Ho) ranged from 0 .438 ue between paired populations was from -0 .015 64 to 0 .252 47 ,and the polymorphic information content in the population ranged from 0 .528 -0 .857 ,showing a high polymorphism .Hierarchical AMOVA calculations showed that inter-individual variation of the snails occupied 88 .4% of the total variations .Cluster analysis revealed that the three ribbed shell population in Munu Kou Village ,Hengti Village and Yixing Village first clustered to the three light shell population in Mashizizu Village , Mingzhu Village and Tuqiao Village ,then clustered to the light shell population in Tuanshan Village and Jiama Cao Village with the two shallow rib population in Desheng Village and Tianmu He Village .Under the different landscape environment of Songzi Area ,there were different shells presenting on the morphology of O .hupensis .Although there was a rich diversity on O .hupensis of Songzi City ,the genetic differences mostly present in individuals .Different groups didn’t show the significant genetic differentiation among the different shell morphology of O .hupensis .
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<p><b>OBJECTIVE</b>To study the expression of RAS protein in human glioma tissues and its influence on tumor growth.</p><p><b>METHODS</b>RAS protein expression in glioma tissues was determined by immunohistochemical (IHC) staining. Subsequently, MTT cell proliferation assay, flow cytometry and Western blotting were used to assay U251 cells with reduced RAS expression.</p><p><b>RESULTS</b>The expression of RAS in glioma was increased and strongly correlated with pathological grade. Downregulation of RAS resulted in glioma cells growth suppression and increased apoptosis.</p><p><b>CONCLUSION</b>The expression level of RAS protein in human glioma was increased. Downregulation of RAS can inhibit glioblastoma cell growth through the RAS signal pathway.</p>
Subject(s)
Humans , Brain Neoplasms , Genetics , Metabolism , Pathology , Cell Growth Processes , Genetics , Cell Line, Tumor , Down-Regulation , Glioma , Genetics , Pathology , Immunohistochemistry , ras Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the efficacy and safety of total glucosides of paeony capsule (TGPC) in patients with mild and moderate alopecia areata.</p><p><b>METHODS</b>A total of 86 outpatients were randomly allocated into two groups of TGPC (treatment, 44 cases) and compound glycyrrhizin tablet (control, 42 cases). The treatment group was given oral TGPC, three times daily and 600 mg per time; the control group was given oral compound glycyrrhizin tablets, three times daily and 50 mg per time. In addition, both groups were given 10 mg of vitamin B(2) and tapped the bold patches with massage. The treatment course was three months for both groups. Peripheral blood T-cell subsets (CD3(+)CD4(+), CD3(+)CD8(+), Th, Ts, Th/Ts) of 10 patients randomly selected from each group respectively were tested before and after three months of treatment. The effectiveness and adverse reaction of all cases were observed each month. The safety was evaluated according to the incidence rate of adverse reaction.</p><p><b>RESULTS</b>In the treatment group, the cured and markedly effective rate was 36.36% (16/44), 50.00% (22/44) and 68.18% (30/44) at the end of first, second and third month of treatment, respectively, and the incidence rate of adverse reaction was 13.64% (6/44). In the control group, the cured and markedly effective rate was 38.10% (16/42), 57.14% (24/42) and 71.43% (30/42), respectively, and the incidence rate of adverse reaction was 16.67% (7/42). The cured and markedly effective rate and the incidence rate of adverse reaction were similar in both groups (P>0.05). TGPC and compound glycyrrhizin tablet can inhibit CD3(+)CD4(+) and CD3(+)CD8(+), and decrease the ratio of Th/Ts (P<0.05).</p><p><b>CONCLUSION</b>TGPC is effective and safe in the treatment of alopecia areata.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Alopecia Areata , Drug Therapy , Allergy and Immunology , Capsules , Glucosides , Therapeutic Uses , Glycyrrhizic Acid , Therapeutic Uses , Lymphocyte Subsets , Allergy and Immunology , Paeonia , Chemistry , T-Lymphocytes , Allergy and Immunology , Tablets , Treatment OutcomeABSTRACT
<p><b>BACKGROUND</b>Invasion growth is the most characteristic biological phenotype of glioblastoma, but the molecular mechanism in glioma cell invasion is poorly understood. Recent data have showed that microRNA plays an essential role in tumor invasion. Our study aimed to explore the mechanism of miR-7 involved in the control of glioblastoma cell invasion.</p><p><b>METHODS</b>Glioma cell invasion was evaluated by transwell and scratch assays after up-regulation of miR-7 using miR-7 mimics in U87 and U251 cells. Luciferase reporter assay was used to determine focal adhesion kinase (FAK) as a target of miR-7. The levels of miR-7, matrix metalloproteinases (MMP)-2 and MMP-9 mRNA were detected by PCR assay, and the levels of FAK, MMP-2, MMP-9, total and phosphorylation serine/threonine kinase (AKT), and extracellular signal-regulated kinase (ERK) 1/2 were measured by Western blotting analysis.</p><p><b>RESULTS</b>Over-expression of miR-7 inhibited the invasion and migration activity of U87 and U251 cells. And up-regulation of miR-7 reduced FAK protein expression, Further, luciferase reporter assay showed that miR-7 modulated FAK expression directly by binding 3'UTR of FAK mRNA. In addition, miR-7 repressed p-ERK1/2 and p-AKT level, MMP-2 and MMP-9 expression. Finally, the inverse relationship between FAK and miR-7 expression was certificated in human glioma tissues.</p><p><b>CONCLUSION</b>To our knowledge, these data indicate for the first time that miR-7 directly regulates cell invasion by targeting FAK in glioblastoma and that miR-7 could be a potential therapeutic target for glioblastoma intervention.</p>
Subject(s)
Humans , Blotting, Western , Cell Line, Tumor , Focal Adhesion Protein-Tyrosine Kinases , Genetics , Metabolism , Glioblastoma , Genetics , In Vitro Techniques , Matrix Metalloproteinase 2 , Genetics , Metabolism , Matrix Metalloproteinase 9 , Genetics , Metabolism , MicroRNAs , Genetics , Metabolism , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of bone morphogenetic protein 4 (BMP4) on the proliferation and apoptosis in glioma stem cells.</p><p><b>METHODS</b>Stem cells were isolated from a human glioma cell line U87 by using vincristine and characterized by immunofluorescence assay. Proliferation and apoptosis were determined by soft agar colony assay and flow cytometry; Cyclin D1, Bcl-2 and Bax were detected by Western blot analysis.</p><p><b>RESULTS</b>BMP4 inhibited cell proliferation and promoted apoptosis in U87 glioma stem cells. Moreover, Bcl-2 and Cyclin D1 expression were decreased by BMP4, while Bax level was elevated.</p><p><b>CONCLUSION</b>BMP4 can inhibit U87 glioma stem cells proliferation through downregulating Cyclin D1 level, and promote apoptosis through induction of Bax expression and inhibition of Bcl-2 level. It suggests that BMP4 plays an important role in human glioma stem cell biology.</p>
Subject(s)
Humans , Apoptosis , Bone Morphogenetic Protein 4 , Genetics , Metabolism , Cell Line, Tumor , Cell Proliferation , Cyclin D1 , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , Glioma , Genetics , Metabolism , Neoplastic Stem Cells , Cell Biology , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the potentiality of herpes simplex virus thymidine kinase transduced endothelial progenitor cells (EPC-TK) as angiogenesis-targeting vector in the glioma treatment in vitro and in vivo.</p><p><b>METHODS</b>EPC-TK were mixed with human umbilical vein endothelial cells (HUVECs), U87 or U251 cells at various ratios for ganciclovir (GCV) treatment. The bystander effect was observed by counting the survival cells using MTT assay, and the apoptotic cells were determined by annexin-V and propidium iodide (PI) staining. EPC-TK, EPCs, or phosphate buffered saline (PBS) were injected into the nude mice model of glioma xenograft by tail vein, for the EPC-TK group, EPC group, and PBS group, respectively. And then the changes of tumor volume and tumor vasculature were observed.</p><p><b>RESULTS</b>GCV killed most EPC-TK and reduced the number of other viable cells in a cell:cell ratio-dependent and time-dependent manner. EPC-TK obviously inhibited tumor growth. The tumor volumes on day 21 were 1741.20+/- 576.10 mm(3), 3275.52 +/- 710.86 mm(3) and 3033.09+/-1134.86 mm(3) in the EPC-TK, EPC and PBS group, respectively. EPC-TK also displayed a significant effect on the inhibition of tumor angiogenesis.</p><p><b>CONCLUSION</b>EPC-TK can exert a potent antiglioma effect via inhibiting angiogenesis.</p>
Subject(s)
Animals , Humans , Mice , Angiogenesis Inhibitors , Pharmacology , Antiviral Agents , Pharmacology , Bystander Effect , Cell Transformation, Viral , Physiology , Endothelial Cells , Virology , Endothelium , Genetic Vectors , Glioma , Therapeutics , Mice, Nude , Simplexvirus , Genetics , Thymidine Kinase , Genetics , Transduction, Genetic , Transfection , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of osteopontin silencing on the invasion and apoptosis of U251 cells.</p><p><b>METHODS</b>The invasion, apoptosis and levels of uPA, MMP-2 and MMP-9 were determined by invasion assay, flow cytometry, Western blot and real-time fluorescence quantitative PCR respectively.</p><p><b>RESULTS</b>Osteopontin small interference RNA (siRNA) inhibited osteopontin expression and cell invasion, promoted apoptosis in U251 cells. In addition, the expression of Bcl-2, uPA, MMP-2 and MMP-9 was decreased, while Bax level was elevated.</p><p><b>CONCLUSION</b>Osteopontin siRNA can inhibit U251 cells invasion via the down-regulation of uPA, MMP-2 and MMP-9 levels, and promote apoptosis through induction of Bax expression and inhibition of Bcl 2 level. It suggests that osteopontin plays an important role in human glioma progression.</p>
Subject(s)
Humans , Apoptosis , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Gene Silencing , Gene Transfer Techniques , Genetic Vectors , Genetics , Metabolism , Glioma , Genetics , Metabolism , Pathology , Lentivirus , Genetics , Metabolism , Neoplasm Invasiveness , Osteopontin , Genetics , Metabolism , RNA, Small Interfering , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the clinical curative effect of Chinese herbal medicine combined with acitretin capsule in treating psoriasis of blood-heat syndrome (P-BH).</p><p><b>METHODS</b>Eighty patients of P-BH were randomly assigned to two groups, 39 in Group A and 41 in Group B. Both was treated with Chinese herbal medicines for clearing heat, cooling blood and removing toxic substance, and acitretin capsule was given to Group A additionally, with 8 weeks as one therapeutic course. The clinical curative effect was compared between groups, and the change of psoriasis activity severe index (PASI) scores before and after treatment was observed.</p><p><b>RESULTS</b>The total effective rate in Group A was 84.2% and that in Group B 68.2%, also showing significance between groups (P<0.01). PASI score lowered significantly after treatment in both groups, showing statistical significance (P<0.01), but no significant difference between groups. Little adverse reaction was found in Group B, while in Group A, the adverse reaction was of even milder degree, which could be alleviated by adjusting the herbal medicine and symptomatic treatment administration.</p><p><b>CONCLUSIONS</b>The effect of Chinese herbal medicine combined with acitretin capsule was superior to Chinese herbal medicine alone in treating P-BH, but the adverse reaction of acitretin capsule could be alleviated by adjusting the herbs used. However, the result is waiting to be verified further by larger samples.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Acitretin , Capsules , Combined Modality Therapy , Diagnosis, Differential , Drug Combinations , Drugs, Chinese Herbal , Hematologic Diseases , Diagnosis , Drug Therapy , Hot Temperature , Keratolytic Agents , Medicine, Chinese Traditional , Methods , Psoriasis , Diagnosis , Drug Therapy , Syndrome , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To construct a recombinant lentivirus RNA interference (RNAi) vector carrying hTERT gene, and to obtain the titer of the lentiviral stock for investigating the expression in the eukaryotic cells and the effect on the hTERT gene silencing in the eukaryotic cells.</p><p><b>METHODS</b>Two complimentary oligos of small interference RNA (siRNA) with hairpin structures targeting the hTERT gene and a negative control were synthesized, then ligated with pLVTHM vector and sequenced. The recombinant vectors were then transfected with viral packaging mix into T293 cells, viral supernatant was harvested to determine the titer. U87 cells infected by virus were harvested and the expression of hTERT, telomerase activity and apoptosis were detected by reverse transcription-PCR(RT-PCR), TRAP assay and flow cytometry separately.</p><p><b>RESULTS</b>Sequencing data showed that the constructed plasmids contained the correct sequences of hTERT siRNA transcript templates. A vector producing cell line T293 was established, and the titer for transfection was obtained. RT-PCR and TRAP flow cytometry analyses demonstrated that hTERT shRNA expression construct could suppress the expression of hTERT and telomerase activity and induce apoptosis.</p><p><b>CONCLUSION</b>A lentivirus RNAi vector targeting hTERT gene was successfully constructed, which decreased the expression of hTERT and telomerase activity effectively and induced apoptosis. It has set up a research platform for the gene therapy of tumors which take hTERT as the target.</p>
Subject(s)
Humans , Base Sequence , Cell Line , Flow Cytometry , Gene Knockdown Techniques , Methods , Genetic Vectors , Genetics , Lentivirus , Genetics , Molecular Sequence Data , Oligodeoxyribonucleotides , Genetics , Metabolism , Plasmids , Genetics , Metabolism , RNA Interference , RNA, Small Interfering , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Telomerase , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study inhibitory efficacy of combined gene therapy for malignant gliomas transfected with antisense human telomerase reverse transcriptase (hTERT)/PTEN in vitro and in vivo.</p><p><b>METHODS</b>To construct two adenovirus recons which contained antisense hTERT and wild-type PTEN respectively with high performance homologous recombination system in bacteria. The two adenovirus recons were transfected into U251 human malignant glioma cells combinedly or respectively in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured by the volume of glioma in nude mice. Telomerase activity was detected by telomeric repeat amplification protocol (TRAP) assay. Expression of hTERT and PTEN protein was detected by Western blotting methods.</p><p><b>RESULTS</b>After transfection in vitro, the growth of U251 cells was inhibited significantly. The inhibitory effect was time-dependent. The strongest inhibition was observed in combined transfection group, at the 6th day, the survival rate was 37.6%, telomerase activity (only 28.8TPG) was inhibited significantly, hTERT protein expression was inhibited significantly too, which was 0.2106, but PTEN protein expression was increased significantly, which was 0.9630. In vivo, the growth of tumors was also effectively inhibited.</p><p><b>CONCLUSION</b>Growth of malignant glioma cells is effectively inhibited after transfection with combined antisense hTERT and PTEN in vitro and in vivo.</p>
Subject(s)
Animals , Humans , Mice , Adenoviridae , Genetics , Apoptosis , Blotting, Western , Brain Neoplasms , Pathology , Therapeutics , Cell Line, Tumor , Cell Proliferation , DNA, Antisense , Genetics , Metabolism , Flow Cytometry , Genetic Therapy , Methods , Glioma , Pathology , Therapeutics , Green Fluorescent Proteins , Genetics , Metabolism , Mice, Nude , Microscopy, Fluorescence , PTEN Phosphohydrolase , Genetics , Recombinant Fusion Proteins , Genetics , Metabolism , Telomerase , Genetics , Metabolism , Transfection , Tumor Burden , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene.</p><p><b>METHODS</b>C6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-I and IGFBP3 expression.</p><p><b>RESULTS</b>Cx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change.</p><p><b>CONCLUSION</b>The mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.</p>