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.

Objective:To study the morphologic and phenotypic characters of a macrophage-tumor vaccine,and to observe the effect of macrophage-tumor vaccine on inducing CTL respose.Methods:The super-structure and the expression of CD14,CD68,CD80,CD86,MHC Ⅱ molecules of macrophage-tumor cells were detected with electron microscope,immunofluorescence staining and flow cytometry respectively.Meanwhile,H22 tumor cells were transplanted to the mice that had been immunized with different tumor vaccines.The weight and volume of tumors,the tumor cell injure rate and the level of LDH in culture supernatant were detected with direct measurement,MTT and selection methods.Results:The macrophage tumor vaccine cells were large cells with an irregular outline,and generally displayed pseudopodium,membrane folding,and vesicles on the cell surface.The predominant cytoplasmic organelles were lysosomes,secondary lysosomes and residual bodies.The percentage of CD14,CD68,CD80,CD86 and MHC Ⅱ positive cells within the differentiated population were 53.90%,98.60%,26.50%,90.20% and 25.40% respectively.The results of experiment in vivo revealed that the tumor forming rate,volume and weight of the group immunized with macrophage-tumor vaccine were much lower than that of control group and the group that were immunized with the macrophages that were induced by liquid paraffin (P0.05),the tumor weight and volume of the group immunized with the macrophage-tumor vaccine were lower than those of the group immunized with inactivated tumor cells(P

The management of the scientific research files is an important basic work at the college,which is of great significance to the research work in both school and the scientific research organizations.However,some researchers and managers are lack of file consciousness and the management system remains unadapted to the new condition.These reasons result in the use of the dossiers inadequate or even absent.Therefore,it seems necessary and important for the perfection of the scientific research file work to take essential measures:establishing a scientific and reasonable file system,improving the file management regulation,and increasing the utilization ratio of the dossiers.

Objective To investigate the relationship between carotid intima-media thickness (IMT) and coronary atherosclerosis plaque through coronary CT angiography (CTA) in asymptomatic individuals.Methods Two hundred and fifty cases with CTA screened were selected in asymptomatic individuals,and according to the inspection result they were divided into no plaque group (detection of no coronary atherosclerosis plaque,180 cases) and plaque group (detection of coronary atherosclerosis plaque,70 cases).All cases were checked carotid IMT with B-ultrasonic.Results ANOVAs analysis showed that age and body mass index (BMI) had significant correlation with coronary atherosclerosis plaque (P ＜ 0.05).x2 test was used to show the smoking and family history of coronary heart disease had significant correlation with coronary atherosclerosis plaque (P ＜ 0.05).Univariate Logistic regression analysis showed that carotid IMT,systolic pressure,diastolic pressure,low density lipoprotein cholesterol,fasting serum glucose and creatinine had significant correlation with coronary atherosclerosis plaque (P ＜ 0.05).Multifactor Logistic regression analysis showed that carotid IMT had correlation with coronary atherosclerosis plaque (P ＜ 0.05).Conclusion There is significant relationship between carotid IMT and coronary atherosclerosis plaque in asymptomatic individuals.

Radix et Rhizoma Rhei, the major usage of which is purgation, is widely used in clinical therapy among different prescriptions. According to present reports, researches on herbal compatibility mechanism were mainly fo-cused on the comparison of content changes of active ingredients before and after. In oder to provide references, this review was made on researches of compatibility mechanism of prescriptions and paired drugs for ingredients of Radix et Rhizoma Rhei from research actuality.

Objective Severe preeclampsia (sPE), which is usually complicated by small-for-gestational-age (SGA) and immature labor , remains a leading cause of maternal and neonatal mortality and morbidity. This study was to investigate the risk factors of SGA in sPE. Methods We retrospectively analyzed 100 cases of sPE, 35 with SGA (the case group) and the other 65 without it ( the control group ) .We conducted single-factor analysis on the general characteristics and potential impact factors of the patients , i-dentified the independent risk factors of SGA using the un-conditional stepwise logistic model , and assessed the value of umbilical arter-y S/D ratio and proteinuria ration in the diagnosis of sPE with SGA with the ROC curve . Results Compared with the control group , the case group had more cases of early-onset sPE ( P =0.010 ), earlier gestational and delivery weeks (P<0.001), lower neonatal weight at birth ( P<0 .001 ) , higher rate of admission to and longer stay in the neonatal ICU (P<0.001), and higher incidence of neo-natal complications (P<0.05).The case group also showed signifi-cant increases in comparison with the control in the umbilical artery S/D ratio (2.95 ±0.75 vs 2.31 ±0.47, P<0.05), frequency of S/D ratio ≥95th percentile (22.86% vs 6.15%, P<0.01), and rate of proteinuria ≥5 g/24 h (42.86% vs 20.00%, P<0.05).The S/D ratio ≥95th percentile (OR=6.02, 95%CI:2.32-16.78) and proteinuria≥5 g/24 h (OR=1.65, 95%CI:1.56-3.01) were found to be the risk factors of sPE with SGA.The area under the curve was 0.852 for the combination of S/D ra-tio and proteinuria ration in the diagnosis of sPE with SGA (P<0.05). Conclusion Umbilical artery S/D ratio≥95th percentile and proteinuria ≥5 g/24 h contribute to the early prediction, prevention, and prognosis of sPE, and is valuable for the diagnosis of sPE with SGA.