BACKGROUND: Previous studies have demonstrated that berberine represses multiple tumors and tumor stem cells, but the effect of berberine on lung cancer stem cells (LCSCs) remains unclear. OBJECTIVE: To explore the effect of berberine on the proliferation and apoptosis of LCSCs and the possible mechanism. METHODS: CD133+ LCSCs were separated from A549 cells by immunomagnetic beads. The effects of different concentrations (0, 2.5, 5, 10, 20, 40 mg/L) of berberine on the proliferation and apoptosis of LCSCs were determined by MTT and flow cytometry analysis, respectively. In order to further affirm the effect of berberine on the proliferation and apoptosis of LCSCs, the expression levels of Ki67, Bax and Bcl-2 protein were detected by western blot. In addition, to investigate the potential mechanism by which berberine exerts regulatory effects on LCSCs, the expression levels of Hedgehog signaling pathway-associated proteins (PTCH1, SHH, Gli-1 and SMO) were determined. RESULTS AND CONCLUSION: After magnetic cell sorting, the content of the CD133+ fraction was enriched up to 84.13%. MTT and flow cytometry assays showed that berberine inhibited proliferation and promoted apoptosis of LCSCs in a concentration-dependent manner. Western blot analysis showed that the expression levels of Ki67, Bcl-2, PTCH1, SHH, Gli-1 and SMO proteins of LCSCs cultured in the medium with 20 mg/L berberine were dramatically decreased compared to the control, while the expression level of Bax protein was markedly increased compared to the control. These findings suggest that berberine may inhibit proliferation and promote apoptosis for LCSCs through the Hedgehog signaling pathway.BACKGROUND: Previous studies have demonstrated that berberine represses multiple tumors and tumor stem cells, but the effect of berberine on lung cancer stem cells (LCSCs) remains unclear. OBJECTIVE: To explore the effect of berberine on the proliferation and apoptosis of LCSCs and the possible mechanism. METHODS: CD133+ LCSCs were separated from A549 cells by immunomagnetic beads. The effects of different concentrations (0, 2.5, 5, 10, 20, 40 mg/L) of berberine on the proliferation and apoptosis of LCSCs were determined by MTT and flow cytometry analysis, respectively. In order to further affirm the effect of berberine on the proliferation and apoptosis of LCSCs, the expression levels of Ki67, Bax and Bcl-2 protein were detected by western blot. In addition, to investigate the potential mechanism by which berberine exerts regulatory effects on LCSCs, the expression levels of Hedgehog signaling pathway-associated proteins (PTCH1, SHH, Gli-1 and SMO) were determined. RESULTS AND CONCLUSION: After magnetic cell sorting, the content of the CD133+ fraction was enriched up to 84.13%. MTT and flow cytometry assays showed that berberine inhibited proliferation and promoted apoptosis of LCSCs in a concentration-dependent manner. Western blot analysis showed that the expression levels of Ki67, Bcl-2, PTCH1, SHH, Gli-1 and SMO proteins of LCSCs cultured in the medium with 20 mg/L berberine were dramatically decreased compared to the control, while the expression level of Bax protein was markedly increased compared to the control. These findings suggest that berberine may inhibit proliferation and promote apoptosis for LCSCs through the Hedgehog signaling pathway.

Objective: To express glycosyl-phosphatidylinositol (GPI) modified Met- RANTES fusion protein on Chinese hamster ovary (CHO) cells and to develop a novel immunosuppressant GPI anchored form of Met-RANTES. Methods: The eukaryotic expression vector PEF/GPI-Met-RANTES were constructed and transfected into CHO cells by electroporation. The transfectants were selected with methotrexate (MTX). Expression of the recombinant protein was assessed by flow cytometric analysis, cell immunofluorescence staining and immunogold electron microscopy. Results: The chimeric molecules of GPI anchored form of Met-RANTES including the whole reading frame were constructed, and the sequence was identical to the designed sequence. GPI anchored form of Met-RANTES was stably expressed on CHO- DHFR- cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES may be used as novel immunosuppressant for suppressing reaction in graft rejection.

Objective To study the expression of telomerase and its possible clinical significance in patients with condyloma acuminatum,bowenoid papulosis,Bowen's disease and vulvar squamous cell car- cinoma.Methods The human telomerase reverse transcriptase (hTERT) was detected by immunohisto- chemistry technique in all patients.Histopathological analysis was done in patients with bowenoid papulosis or Bowen's disease.Results There was a significant difference in the expression of hTERT among normal tissue,condyloma acuminatum and squamous carcinoma,and the expression rate increased successively from normal control to squamous carcinoma.In bowenoid papulosis,the expression of bTERT was significantly higher as compared to that in condyloma acuminatum,significantly lower as compared to that in squamous carcinoma,and was as the same as that in Bowen's disease.Histopathologically,the degree of cell atypia was significantly higher in Bowen's disease than that in bowenoid papulosis.Conclusions The expression of telomerase increases successively from normal skin,benign tumors to malignant carcinomas,which sug- gests that the expression of telomerase may play an important role in both cell proliferation and immortaliza- tion.

Background The rat model of N-methyl-N-nitrosourea (MNU)-induced photoreceptor cell apoptosis is often used to study retinal degeneration.But the changes in the gene expression patterm in retinal degeneration in rats have not been reported.Objective This study was undertaken to investigate regulation of gene expression in the retina of MNU-induced retinal degeneration in rats by performing microarray analysis of retinal RNA.Methods Fifty 6-week-old SD rats were numbered and randomized into the normal group and the model group.The retinal degeneration model was established by a single hypodermic injection of 40 mg/kg of MNU,and the rats in the normal group received equivalent volume of physiological saline in the same way.The rats were sacrificed 12 hours or 24 hours after injection.Retinal sections from the right eyes were prepared for the measurement of the retinal thickness by histopathological examination,and retinas from the left eyes were used to confirm the differential gene expression as detected by microarray (normal group and 12 hours model group).Genes exhibiting changes in expression by ≥2.0 folds were further confirmed using real-time PCR.Results The whole thickness of the retina declined in the rats from the 24 hours model group compared to the normal group and 12 hours model group (t =9.926,P=0.002;t=2.736,P=0.028).The thickness of the outer nuclear layer was (26.58±2.90) μm in the 24 hours model group,showing a significant decrease in comparison with (38.11 ± 1.01) μm in the normal group and (35.07t3.03) μm in the 12 hours model group (t=6.028,P=0.009;t=6.839,P=0.006).However,there was no significant difference in retinal thickness between the normal group and the 12 hours model group (whole thickness:t=1.541,P=0.324;outer nuclear layer thickness:t=2.040,P=0.134).Microarray analysis of the rat genes showed that out of 17 000 genes,142 genes involved in biological process and 94 genes involved in molecular functions were differentially expressed,where most of them participate in the mitogen activated protein kinase signaling pathway,Tolllike receptor signaling pathway and apoptosis pathway.Real-time PCR analysis demonstrated that the expression of CCL2,IL-1b,CCL3,c-fos,c-myc,p53 and MMP3 were consistently up-regulated,conforming with the results from microarray analysis.Conclusions The changes in gene expression pattern appear in the early stage of MNUinduced retinal degeneration.These microarray results provided clues to understanding the molecular pathways underlying photoreceptor degeneration and indicating the directions for future studies.

ObjectiveTo explore the influences of age and duration of status convulsivus （SC） on mitochondrial membrane potential (△Ψm） in hippocampus. MethodsConvulsive seizures for 30 min or 3 h (30 min SC or 3 h SC) were induced in 80 infant (20 d after birth) and 80 adult Wistar rats (IRs ＆ ARs respectively) with lithium-pilocarpine ip. The rats were sacrificed at 6 different time points from the 3rd hour to 7th day after SC termination. The mitochondrial △Ψm in hippocampal cells was determined with flow cytometry. ResultsThe mitochondrial △Ψm in hippocampal cells started to decrease at the 3th hour after SC in both IRs and ARs. The bottom level was reached at the 6th hour after SC ［(6.08±0.43) in IRs and (5.70±0.63) in ARs ) ］. Both of them were significantly lower than that of control group (P<0.01） and began to increase at 12th hour after SC. On the 7th day after 30 minutes SC, the level of mitochondrial △Ψm in IRs increased to the level of control, while the level in ARs was still lower than that of control (P<0.05）. At the 3rd hour, the 3rd and the 7th day after SC, the levels of mitochondrial △Ψm in IRs were obviously higher than those in ARs. Compared with the same time point after 30 min SC, the levels of mitochondrial △Ψm at the 3rd and the 6th hour after 3 h SC were much lower in different age groups (P<0.05）. Except the effect of the age-related difference, there was a positive correlation between the duration of SC and the changes of mitochondrial △Ψm in partial correlation analysis （r=0.71，P<0.05）. ConclusionSevere seizure could induce the mitochondrial △Ψm decreased in hippocampus. Age and duration of SC were important factors associated with the mitochondrial △Ψm decrease. There may be an internal protective response against brain damage in premature brain.

Acupuncture Therapy ; instrumentation ; Adult ; Female ; Humans ; Lactation ; Mastitis ; physiopathology ; therapy ; Young Adult

Anemia, Refractory ; Humans ; Male ; Middle Aged ; Thrombocytosis

As the major commercial source of natural rubber,Hevea brasiliensis attracts much attention.However,the heterozygous nature,long breeding cycle are strong limitations for conventional breeding.While genetic engineering,which can be used to widen the germplasm base and produce desirable agronomic traits quickly and efficiently,offers a viable alternative approach to complement traditional breeding.Comprehensive analysis indicates that in the past two decades,with calli derived from immature anther or integumental tissues of immature fruit as receptors,both biolistic and Agrobacterium-mediated transformation methods were employed for developping rubber genotypes with improved latex yield,tolerance to tapping panel dryness syndrome,producing high-value recombinant proteins,etc.Being recalcitrant to tissue culture,the transformation efficiency of Hevea is comparatively low,and the procedures are still needed to optimize.Finally,breeding objectives and strategies to improve transformation efficiency were also proposed in the review.

Acoustic Stimulation ; Animals ; Corpus Striatum ; physiology ; Estrogens ; metabolism ; Estrus ; physiology ; Evoked Potentials, Auditory ; physiology ; Female ; Rats ; Rats, Sprague-Dawley

Rheumatoid arthritis (RA) is a kind of chronic, progressive, multiple, invasive autoimmune disease with two chief cclinical manifestations arthrosynovitis and ex-arthrosis, easy to occur in middle-aged women, also occur in children and the elderly, is characterized by progressive and break out repeatedly. RA pathogenesis is complex, there is no special treatment, used in treatment of R drug varied, new drugs and new therapies also emerge in endlessly, main including non-steroidal anti-inflammatory drugs (NSAIDs), slow action anti-rheumatism medicine (SAARDs), glucocorticoids (GCs), biological agent, traditional Chinese medicine and traditional Chinese medicine preparations, domestic market for rheumatoid main drug treatment are NSAIDs, SAARDs, GCs, traditional Chinese medicine and traditional Chinese medicine preparations. Traditional Chinese medicine and traditional Chinese medi- cine preparations for the treatment of RA have its unique advantages, show the characteristics of overall adjustment, multi-level and multiple targets, and also can alleviate and against side effects of western medicine. In recent years, more and more get people's atten- tion. This paper reviewed the research progress and treatment features of commonly used therapeutic agents for the treatment of RA in recent years, which provides reference and basis for future medicine anti-RA.
Animals ; Arthritis, Rheumatoid ; drug therapy ; Biological Factors ; adverse effects ; therapeutic use ; Drug Discovery ; methods ; Glucocorticoids ; adverse effects ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; adverse effects