Objective To construct a library of long serial analysis of gene expression (LongSAGE) and investigate the characteristics of gene expression in the yeast phase of C. albicans. Methods A LongSAGE library was constructed with long serial analysis of gene expression (LongSAGE). Results A total of 17 773 tags, representing specific 7 333 transcripts, were extracted from 1 000 sequence files in the yeast phase of C. albicans. Tags matching single gene accounted for 16.65%, tags matching multiple genes accounted for 6.59%, tags for hypothetical protein accounted for 16.27%, tags for genome and cosmid accounted for 1.64%, and novel tags accounted for 58.86%. Eleven tags in 33 tags whose expression frequencies exceeded 35 may be new expression sequences. Conclusion LongSAGE is not only a powerful method in investigating the gene expression profiling, allowing the analysis of the expression of thousands of transcripts in a quantitative manner without prior sequence information, but also in finding new genes.

Objective To explore the expression of SLAM gene in the CD4+ and CD8+ T cells from SLE patients.Method The CD4+ and CD8+ T cells from peripheral blood of 20 SLE patients and 10 healthy blood donors were separated using magnetic cell sorting system(MACS).We detected the expression of SLAM mRNA in CD4+ and CD8+ T cells by RT-PCR.Results The CD4+ and CD8+ T cells from SLE patients showed significantly higher SLAM expression than those from healthy blood donors.Conclusion The SLAM signal pathway may play an important role in contributing to the abnormal activation of T cells in SLE patients.

Objective To explore the expression of caspase-3 and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors in the CD4+ and CD8+ T cells of systemic lupus erythematosus (SLE)patients. Methods The CD4+ and CD8+T cells from peripheral blood of 20 SLE patients and 10 healthy volunteers were separated using magnetic cell sorting system (MACS), reverse transcription-polymerase chain reaction (RT-PCR) was used to test the expression of caspase-3 and TRAIL receptors in CD4+ and CD8+T cells of SLE patients and healthy volunteers. Results The CD8+T cells from SLE patients had significantly higher caspase-3 (P=0.003) and TRAIL-R2 (P=0.024) expression than those from healthy volunteers. Conclusion The TRAIL-R2 signal pathway may be a possible important pathway to the apoptosis of T cells in SLE patients.

Objective To investigate the abnormality of CD4 and CD8 subsets of T lymphocytes in the peripheral blood of systemic lupus erythematosus patients. Methods The CD4 + and CD8 + T lymphocytes from 20 SLE patients and 10 healthy controls were separated using magnetic activated cell sorting (MACS). Results The numbers of CD4 + T cells in the peripheral blood and CD4 +/CD8 + ratio of SLE patients were significantly lower than those in healthy volunteers(CD4,P=0.001;CD4/CD8 ratio,P=0.046). Conclusion There exist abnormalities in the CD4 and CD8 subsets of T lymphocytes in the peripheral blood of SLE,especially the decreased numbers of CD4 + T cells.

Objective To investigate the distribution of air microorganisms in soldies′living quarters of one under-ground tunnel during airtight survival training .Methods Anderson sampling was carried out in Area A and Area B ( living quarters) and Area C (a toilet) at designated time and place.After sampling, the culture and identification of microorgan-isms were finished .Results ①In the living quarters of the whole underground tunnel , the total number of bacteria ranged from 125 to 37 800/m3,but it was 2692, 1844 and 2199/m3, respectively in Area A, B and C.The mean number of bac-teria was 2245/m3 .The number of fungi ranged from 0 to 10 017/m3 .The total number of fungi of Area A , B and C was 1064 , 883 and 1011/m3 .The mean number was 986/m3 .②The number of bacteria in the living areas presented three fea-tures:the total bacteria showed overall three peaksin Area A and B, buttwin peaksin Area C.In Area A and B, the number of bacteria exhibited low colony at first but fluctuated heavily later .In Area C, it decreased gradually to the mini-mum,and the fungi showed a wavy and irregular trend .③The air microbial species included cocci ( Micrococcus, Coriolis of Staphylococcus aureus, S.epidermidis and S.equorum) , bacilli ( Acinetobacter baumannii, A.lwoffii and Alcaligenes faeca-lis),and fungi (Mucor and Saccharomyces).Conclusion Although the content of microorganisms was up to the military hygiene standards , it was higher than in the same kind of tunnels .Themulti peakphenomenon of microbial distribution suggests that the change of air microorganisms in tunnels has its own characteristics .Most of the air microorganisms are con-ditioned pathogens that may cause illness if they are not under control .

Objective To investigate the effect of nimodipine (ND) injecting into cisterna magna on the mitochondrial pathway in hippocampus in rabbit model of subarachnoid hemorrhage (SAH). Methods Eighteen New Zealand white rabbits were randomly allocated to Sham group, SAH group and ND group, six in each group. All the animals underwent operation under anaesthesia. One mL/kg autologous non-heparinized arterial blood was injected into cisterna magna in SAH group and ND group, and the same dosage of saline was injected into cisterna magna in Sham group. Thirty minutes after injection, 1 mg/kg nimodipine was injected into cisterna magna in ND group, and equal-volume of saline was injected into cisterna magna in Sham group and SAH group. All the animals were assessed for the grade of food intake and neurological impairment, and rats were killed 72 hours after SAH. Their hippocampus were processed for detecting the expressions of Bcl-2, Bax, Caspase-3 and Cyt-C mRNA by qRT-PCR. The protein expressions of Caspase-3 and Cyt-C were detected by Western blot assay. Results Compared with the Sham group, there were lower grade of food intake, varying degrees of neurological impairment and lower ratio of Bcl-2/Bax, while the mRNA levels of Bcl-2, Bax, Caspase-3 and Cyt-C and protein levels of Caspase-3 and Cyt-C showed elevated expressions in SAH group and ND group (P<0.05). Compared with SAH group, there were no significant differences in the score of food intake and neurological impairment in the ND group ( P>0.05). There were higher ratio of Bcl-2/Bax and expression levels of Bcl-2 mRNA and lower expression levels of Bax mRNA, Caspase-3 and Cyt-C mRNA and proteins in ND group than those in SAH group (P<0.05). Conclusion Nimodipine plays a protective role in inhibiting the activity of mitochondrial pathway in hippocampus after subarachnoid hemorrhage.

Rheumatoid arthritis (RA) is a chronic autoimmune disorder primarily occurred in small joints.Our previous studies suggested that sufficient lymphatic drainage was favorable for the recovery of RA.This study aimed at exploring the effect of Er Chen combined with Tao Hong Si Wu Tang (ECSWT) on RA in TNF transgenic (TNF-Tg) mice.Ten-week old TNF-Tg mice and WT littermates were detected with indocyanine green-nearinfrared (ICG-NIR) lymphatic imaging system before and after accepting the 12-week intragastric administration of ECSWT.All ankle joints were assessed by micro-CT scanning.According to three dimensional images of Micro-CT,it was found that the ankle joints in the TNF-Tg group were much eroded compared with the control group.The bone mass and structure were protected after the treatment of ECSWT.ICG-NIR results showed that lymphatic clearance rate of the TNF-Tg group decreased compared with those of the control group.In comparison with the TNF-Tg group,ECSWT promoted the repair of lymphatic function.Compared with the control group,the pulse value of the TNF-Tg group declined;while this condition could be rescued by ERSWT.In conclusion,ECSWT mitigated bone erosion of astragalus bone area in TNF-Tg mice in contrast to the saline mice,while promoted the pulse value and clearance of lymphatic vessels afferent from footpad to popliteal lymph node,implying that ECSWT was a promising agent for treating RA through its promoting lymphatic drainage function effects.

Objective: To study the aortic root imaging and clinical application in 15 patients with transcatheter aortic valve implantation (TAVI).
Methods: A total of 15 patients with severe aortic valve stenosis received TAVI in our hospital from 2011-03 to 2013-11 were studied. The CT scan and transthoracic echocardiography were conducted to measure the aortic root anatomy and the differences of annulus size between CT and echocardiography were calculated. The prosthetic valves were selected based on CT measurement. The pre-operative accuracy of measurement was evaluated by the follow-up study at 6 months after operation.
Results: The CT measured pre-operative aortic annulus short diameter was (21.5 ± 2.4) mm, long diameter was (27.3 ± 2.7) mm, the average inner diameter was (24.4 ± 2.4) mm, left ventricular out lfow (LVOF) tract long diameter was (28.3 ± 4.5) mm, the average inner diameter of LVOF was (24 ± 3.5), ascending aorta diameter was (35.3 ± 4.4) mm. The Venus Medtech A-Valve implanted in 8 patients with #26 and in 7 patients with #29. The average inner diameter of aortic annulus measured by CT was larger than transthoracic echocardiography, P<0.001. During 6 months follow-up period, no patients had aortic root rupture, coronary obstruction, moderate and severer aortic and peri-aortic regurgitation. There were 4 patients with atrio-ventricular block and received permanent pacemaker implantation.
Conclusion: There is a difference for aortic annulus size by CT and transthoracic echocardiography measurements. CT may presisely assess the aortic root morphology and provide strong support for TAVI.

Objective To compare the effects of two methods of nimodipine administration on mitochondrial pathway in the hippocampus of rabbit models of subarachnoid hemorrhage.Methods Twenty-four New Zealand white rabbits were randomly allocated to sham-operated group,subarachnoid hemorrhage (SAH) group and nimodipine introvenous injection (ND1) group,and nimodipine intracistemal administration (ND2) group (n=6).All animals underwent operation under anaesthesia;one mL/kg autologous nonheparinized arterial blood was injected into cisterna magna in SAH group,ND1 and ND2 group,and one mL/kg saline into cisterna magna in sham-operated group.Thirty minutes after SAH,0.5 mL/kg 0.2％ nimodipine was injected into cisterna magna in ND2 group,and equal-volume saline into cisterna magna in sham-operated group,SAH group and ND1 group.While 0.5 mL/kg 0.2％ nimodipine via introvenously injection was performed in the ND1 group,and equal-volume saline via introvenously injection into the sham-operated group,SAH group and ND2 group.All animals were assessed for the grading of food intake and neurological impairment 72 h after SAH,and then,they were scarified;their hippocampi were processed for detecting the mRNA and protein expressions of Caspase-3 and Cyt-c by using real time-PCR and Western blotting.Results The differences of food intake and neurological impairment between the four groups were statistically significant 72 h after SAH (H=16.664,P=0.001;H=15.411,P=0.001);according mean rank,the food intake and neurological impairment in the ND2 group were decreased as compared with those in the ND2 group.The mRNA and protein expression levels of Caspase-3 and Cyt-c among the four groups were statistically different (P＜0.05).As compared with those in the sham-operated group and SAH group,the mRNA and protein expression levels of Caspase-3 and Cyt-c were significantly higher in the ND1 and ND2 groups (P＜0.05);As compared with ND1 group,ND2 group had significantly lower mRNA and protein expression levels of Caspase-3 and Cyt-c (P＜0.05).Conclusion Intracistemal administration ofnimodipine could decrease mRNA and protein expressions of Caspase-3 and Cyt-c and inhibit activation of mitochondrial pathway in hippocampus in rabbit models of SAH.

OBJECTIVETo investigate the role of bone morphogenetic protein 4 (BMP4) in culturing induced pluripotent stem cells (iPSCs) and the related signal pathways.

METHODSWe amplified the mature peptide of BMP4 from the placenta through RT-PCR, and IgK secretion peptide was ligated to the N-terminal of BMP4 mature peptide. The recombinant plasmid pPYCAG-IgK-BMP4 was transfected into 293T cells and screened with puromycin, and the positive clones for expressing BMP4 were verified by cell immunofluorescence and Western blotting. To test the bioactivity of BMP4, iPSCs were cultured in the medium supplemented with leukemia inhibitory factor (LIF) plus the supernatant containing BMP4, and the cell phenotype, cell differentiation capacity into lineages of the 3 germ layers and expression levels of pluripotency-associated genes were investigated.

RESULTSSmad1 was phosphorylated by BMP4 from the culture medium. iPSCs cultured in the medium supplemented with LIF plus the supernatant containing BMP4 for 3 passages maintained the phenotype of stem cells with the expression levels of pluripotency-associated genes not affected. These iPSCs also maintained the capacity to differentiate into cell lineages of the 3 germ layers.

CONCLUSIONBMP4 can be efficiently expressed in mammalian cells to maintain the multipotent differentiation capacity of the iPSCs in in vitro culture.

Animals ; Bone Morphogenetic Protein 4 ; genetics ; Cell Differentiation ; genetics ; Cells, Cultured ; Female ; Gene Expression ; HEK293 Cells ; Humans ; Immunoglobulin kappa-Chains ; genetics ; Induced Pluripotent Stem Cells ; cytology ; Mice