Purpose:To study the expression of CD44v6 protein in breast carcinoma and its prognostic significance. Methods:100 cases of formalin-fixed paraffin-embeded female breast invasive ductal carcinoma tissues were retrospectively analyzed using EnVision~(TM) immunohistochemical method with the monoclonal antibody CD44v6.Statistical analysis was based on the Log-rank test and Cox analysis.Results:Sixty-six of 100 cancer tissues expressed CD44v6.Positive staining was mainly on the cell membranes.There was significant correlation between CD44v6 immunoreactivity and lymph node me- tastasis and TNM stage.The 5-and 10-year survival rates were 82.76% and 78.37% of patients with CD44v6 low-expres- sion tumors,and 64.1% and 49.88% of those with CD44v6 high-expression tumors,respectively;the difference between the two groups of patients was significant(P

Objective To investigate the effects of siRNA targeting CD 147 gene on the expression of CD147 in melanoma cell line A375, and on proliferation of these cells. Methods Previously prepared recombinant plasmid pSUPER/CD147 siRNA was used. In this study, the recombinant was transfected into the A375 cells. The mRNA expression of CD147 was measured by semi-quantitive RT-PCR, the proliferation of the cells by MTT assay. Results After the transfection with pSUPER/CD147 siRNA1 and pSU-PER/CD147 siRNA2, the mRNA expression of CD147 in the A375 cells was significantly down-regulated by 90.81% (P

ObjectiveTo study the morphological and functional changes of thyroid in lactating rats and their offspring in iodine deficiency and iodine excess animal models.MethodsOne hundred and twenty Wistar rats(30 males and 90 females) were selected.Based on their body weight,the 90 females were stratified and randomly divided into five groups( 18 in each group):low iodine group 1 and group 2(fed with low iodine feed and deionized water containing iodine of 0,5 μg/L) ; high iodine group 1,group 2 and control group(feed with normal diet and deionized water containing iodine of 3000,10 000,50 μg/L).After fed for 3 month,all female rats were mated with males in a ratio of 3 ∶ 1.After birth for 10 days,8 female rats and their offspring in each group were sacrificed.Changes of thyroid were observed by naked eyes.The thyroid weight was measured and pathological changes of thyroids were observed under light microscope.Results①Absolute and relative weight of lactating rats thyroid in low iodine group 1 and group 2 [ (92.02 ± 24.40 ),(77.11 ± 23.32 )mg,(0.509 ± 0.072),(0.384 ± 0.089) mg/kg] were much higher than that of control group[ (17.41 ± 9.25)mg,(0.102 ± 0.016)mg/kg,all P＜ 0.05].Absolute and relative weight of lactating rats thyroid in high iodine group 1 and group 2[(8.22 ± 0.41 ),(9.42 ±0.43)mg,(0.047 ± 0.006),(0.035 ± 0.005)mg/kg] were lower than that of control group(all P ＜ 0.05).Absolute and relative weight of lactating rats and their offspring thyroid was decreased with increase of iodide intake in the diet.②Thyroid enlargement of lactating rats in low iodine group 1 and group 2 was evident,but that of high iodine group 1 and group 2 was not.③Epithelial cell hyperplasia and smaller follicular cavity were observed in low iodine group 1 and group 2 under light microscope.Epithelial cell deformation and mostly flat were observed in high iodine group 1 and group 2.ConclusionsThyroid morphology is changed with iodide intake in the lactating rats and their offspring,and the changes are consistent between female rats and their newborns.

Objective To observe the expression of Na+/I- symporter(NIS) in cultured lactating mammary cells with different levels of iodine and the effect of tumor necrosis factor-α(TNF-α). Methods Original generation of mouse lactating mammary cells cultured in vitro were divided into low iodine group Ⅰ (LI-Ⅰ), low iodine group Ⅱ (LI-Ⅱ), adequate iodine group(AI), high iodine group Ⅰ(HI-Ⅰ), and high iodine group Ⅱ(HI-Ⅱ). Cells were cultured in DEME/F12 culture medium for 24 h with different concentrations of iodine (0,5,50,3000 and 10 000 μg/L, respectively), and TNF-α( 10-2 mg/L) was added to some of cultured cells for 24 h. The expression of NIS mRNA of lactating mammary cells was determined by real-time quantitative PCR and the expression of NIS protein was detected by In-Cell Western. Results In iodine alone group, the expression of NIS mRNA in LI-Ⅰ group [ (64.66 ± 14.99) x 10-4] was higher than that of AI group[ (22.76 ± 7.36) × 10-4, P ＜ 0.01 ]; HI-I group[ (10.18 ±3.53) × 10-4] and HI-Ⅱ group[ (8.59 ± 2.89) × 10-4] were lower than that of AI group(all P ＜ .0.05); With increased iodine concentration, the expression of NIS mRNA decreased. The expression of NIS mRNA in LI-Ⅰ group [(2.72 ± 0.45) × 10-4], LI-Ⅱ group[ (2.69 ± 0.68) × 10-4] and AI group[(1.80 ± 0.67) × 10-4] with iodine plus TNF-o were all lower than that of LI-Ⅰ group, LI-Ⅱ group[ (29.82 ± 4.47 ) × 10-4], and AI group without TNF-α (all P ＜ 0.01). In iodine plus TNF-α, the expression of NIS mRNA in HI-Ⅰ group[(6.58 ± 2.87) × 10-4] and HI-Ⅱ[(7.04 ± 1.36) × 10-4] group were all higher than that of AI group(all P ＜ 0.05); With increased iodine deficiency or iodine excess, the expression of NIS mRNA increased. With increased iodine concentration, the expression of NIS protein decreased in iodine alone group. The expression of NIS protein in iodine plus TNF-α was all lower than that in iodine alone group. In iodine plus TNF-α, the expression of NIS protein increased in both iodine deficiency and iodine excess conditions. Conclusions Iodine may decrease the expression of NIS mRNA and protein of lactating mammary cells. The expression of NIS mRNA and protein of lactating mammary cells was inhibited by TNF-α under different levels of iodine.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

0.05).Conclusion CYPIA1 MspⅠ polymorphism is not related to the susceptibility to colorectal carcinoma.

Objective:To explore the clinical value of remote ECG consultation for primary medical institutions. Methods:Remote ECG consultation data for medical institutions of different levels in Hubei Jianli county were sta-tistically analyzed.Results:In county hospitals of this area,there were 1334 cases of ECG consultation,the positive rate was 41.00% (547/1334),and the three abnormal ECG types with highest incidence rate were arrhythmia (27.29%),left ventricular hypertrophy (LVH,9.15%)and ST-T change (2.62%)in order;in rural health rooms of this area,there were 723 cases of ECG consultation,the positive rate was 55.19%(399/723),and the three ab-normal ECG types with highest incidence rate were arrhythmia (34.02%),ST-T change (11.07%)and LVH (5.39%)in order.Conclusion:The remote ECG consultation is easy to perform.It can rise the detection rate of transient abnormal ECG events in rural area,where is lack of ECG equipment and personnel,so it is worthy of ex-tending.

AIM To investigate the anti-neuroinflammation effects of 4-hydroxybenzyl aldehyde (4-HBAL) from Gastrodia elata Blume on acute cerebral ischemic injury in rats and its nechanism of action.METHODS The rat model of acute cerebral ischemic injury was induced by injecting arachidonic acid via intracarotid artery.Brain tissue samples were taken from the animals 3 h after the model of acute cerebral ischemic injury.Tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were detected in brain tissue to evaluate the effects of 4-HBAL in vivo.Lipopolysaccharid (LPS)-induced activation of BV-2 microglia cells model was used to explore the anti-neuroinflammation mechanism of 4-HBAL.RESULTS The experimental results showed that 4-HBAL had a significant protective effect on acute cerebral ischemic injury.It could significandy decrease the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β),and obviously inhibit the production of nitric oxide (NO),prostaglandin E2 (PGE2) and TNF-α in LPS-stimulated BV-2 cell,and increase the production of interleukin-10 (IL-10) and transforming growth factors-β (TGF-β) in BV-2 cell.CONCLUSION The mechanism of 4-HBAL may be related to the suppression of the excessive activation of microglia after cerebral ischemia and the promotion of the transformation of microglia into anti-inflammatory phenotype.

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics