Objective To investigate the expression of hepatocyte growth factor activator inhibitor- 1 (HAl-1) gene in the tissues of renal cell carcinoma (RCC) and the correlation with tumor angiogenesis.Methods The expression of HAI-1 gene was detected by semi-quantitative RT-PCR in the tissues of 56 cases of RCC,corresponding adjacent non-cancer tissues and 12 nomad renal tissues,the expression of CD34was detected by immunohistochemistry SP method in the above-mentioned organizations,and counted the microvessel density (MVD),analyzed the relationship between the expression of HAI-1 gene in RCC and clinicopathological parameters,MVD.Results The positive expression rate of HAI-1 gene in RCC tissues,the adjacent non-cancer tissues and normal renal tissues were 58.93％(33/56),76.79％(43/56) and 91.67％(11/12),respectively.The expression of HAI-1 gene was a negative correlation to the Fuhrman histological grade,Robson clinical stage (r ＝ -0.617,-0.432,P ＜ 0.01 or ＜ 0.05 ).The positive expression rate in lymph node metastasis was significantly lower than that in without lymph node metastasis [ 35.29％(6/17) vs.69.23％(27/39),P＜ 0.05 ].The MVD in RCC tissues was significantly higher than that in the adjacent noncancer tissues and normal renal tissues(P＜ 0.05).The expression of HAI-I gene in RCC tissues was negative correlated to the MVD (r ＝ -0.516,P ＜ 0.01 ).Conclusions There is a certain relationship between the missing expression of HAI-1 gene in RCC and the development,invasion and metastasis of RCC.The HAI-1 gene deletion can be used as an important molecular marker in the invasion and metastasis of RCC.

OBJECTIVETo investigate effects of the variation of immune function in high humidity environment in different time, and lay a foundation for further study of the related mechanism.

METHODThirty SD rats were divided into 3 groups (n = 10): 20 day group, 40 day group in 90% relative humidity chamber and control group in normal relative humidity. Peripheral blood and spleens were collected to detect the levels of T lymphocyte subsets by Flow Cytometery.

RESULTSIn peripheral blood of the 20 day group rats, the CD3+ %, CD4+ %, CD8+ % and CD4+/CD8+ were 52.91 +/- 6.27, 37.80 +/- 4.11, 14.85 +/- 3.73 and 2.72 +/- 0.82 separately. Expect CD3+ %, they all had significant differences (P < 0.05). In addition, the data of the 40 day group rats showed no diversity in statistics. In spleen, CD8+ % of the 20 day group rats was 6.23 +/- 2.87 with significant differences (P < 0.05) and IgG, IgA and IgM did not change a lot in blood serum of the high humidity groups except C3 of the 20 days group (P < 0.05).

CONCLUSIONIn high humidity environment, the immune function of the rats increased in the initial stage. As time went on, the immune function gradually went to normal level through the self adjustment.

Objective To analyze the clinical features of rectal and perianal inflammatory myofibroblastic tumor and evaluate its diagnosis and treatment.Method Clinicopathological data of 3 cases diagnosed as inflammatory myofibroblastic tumor from January,2005 to June,2011 were retrospectively reviewed.Results Inflammatory myofibroblastic tumor presents as infiltrative growth mass with rich vascularization on CT or MRI,and is difficult to distinguish from hemangioma and other rectal tumors.Preoperative biopsy usually fails to ascertain the entity of mass,and pathological examination of the whole resected specimen with immunohistochemical staining is needed to make final diagnosis.All 3 cases underwent sphincter preserving surgery.One case received a second radical operation 16 months after primary resection because of local recurrence.All patients are followed up to now,with a survival time of 67 months,55 months,and 35 months respectively.Conclusions Rectal and perianal inflammatory myofibroblastic tumor is difficult to diagnose on preoperative imaging examinations or biopsy.Immunohistochemical staining is needed to make final diagnosis.Sphincter preserving surgery with complete tumor removal could achieve long term survival.

The transcript encoding 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) was discovered from the transcriptome data of Huperzia serrata. The transcript contained an open reading frame with length of 1,440 bp and coded 479 amino acids. The full length of HsDXR1 had been cloned using RT-PCR method. Ac-cording the bioinformatic analysis, the molecular weight of HsDXR1 protein was 51.4961 kDa and the pI was 6.44. No signal peptide and transmembrane site was discovered in HsDXR1, and the protein was most likely to be located in chloroplast. HsDXR1 had the same domain similar to the DXR protein of Arabidopsis and Oryza sativa. The expression level of HsDXR1 was most abundantly in H. serrata stem, followed by root and leaf. This study cloned and analyzed HsDXR1 gene from H. serrata for the first time. The result will provide a foundation for exploring the mechanism of terpene biosynthesis in H. serrata plants.

Objective To observe the protein and mRNA expression of LOX-1,eNOS and PPARγ in type 2 diabetic rat aorta,and to investigate the effect of rosiglitazone intervention.Methods Totally 80 Wistar rats (7-week-old male) were randomized into the control group,high fat diet group,diabetic group,and rosiglitazone treatment group (n=20 each).Type 2 diabetes model was developed by intraperitoneal injection with a low dose of streptozotocin,and rats in rosiglization treatment group were treated with rosiglitazone by intragastric administration.After treatment with rosiglitazone for 6 and 12 weeks,animals were sacrificed.Aorta were collected for detecting the protein and mRNA expressions of LOX-1,eNOS and PPARγ,and the differences in expression levels were compared among groups.Results After 6 and 12 weeks,the protein expressions of LOX-1 were up-regulated in diabetic group and rosiglitazone treatment group as compared with control group and high fat diet group (all P＜ 0.01).The protein expression of LOX-1 was down-regulated in rosiglitazone treatment group as compared with diabetic group (P ＜ 0.05).The aorta protein expressions of LOX-1 in high diet group,diabetes group and rosiglitazone treatment group were upregulated after 12 weeks as compared with 6 weeks (all P＜0.01).After 6 and 12 weeks,the aorta protein expressions of eNOS were down-regulated and PPARγ were up-regulated in high fat diet group,diabetic group and rosiglitazone treatment group as compared with control group (all P＜0.01)The aorta protein expression of eNOS was down-regulated and PPARγwas up-regulated in diabetes group as compared with high fat diet group and rosiglitazone treatment group (all P＜0.01).The aorta protein expressions of eNOS in diabetes group and rosiglitazone treatment group were downregulated after 12 weeks as compared with 6 weeks (all P＜0.01).After 6 and 12 weeks,the aorta mRNA expressions of LOX-1 and PPARγ were up-regulated,but the mRNA expressions of eNOS were down-regulated in high fat diet group,diabetes group and rosiglitazone treatment group as compared with control group (all P＜0.05).The aorta mRNA expressions of LOX-1 and PPARγ were up-regulated,but the mRNA expressions of eNOS were down-regulated in diabetes group and rosiglitazone treatment group as compared with high fat diet group (all P＜0.05).The aorta mRNA expressions of LOX-1 and PPARγ were down-regulated,but the mRNA expressions of eNOS were upregulated in rosiglitazone treatment group as compared to diabetic group (all P＜0.01).Conclusions Both hyperglycemia and hyper-lipoproteinemia can induce early coronary atherosclerosis in rats with the abnormal protein and mRNA expressions of LOX-1,eNOS and PPARγ in rat aorta,and the abnormal expressions are more obvious in hyperglycemia combined with hyperlipoproteinemia.Thiazolidinediones can reverse the above abnormal expressions in diabetic rats.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

Nitrite reductases (NiRs) are the key enzymes in the denitrification pathway of the nitrogen cycle. By the catalysis of NiRs, the nitrites are turned into nitric oxides and the nitrogen pollution is decreased in water body. NiRs are divided into two different types based on their prosthetic groups, namely heme-containing nitrite reductases (cd1-NiRs) and Copper-containing nitrite reductases (Cu-NiRs). As all know, Cu-NiRs have trimeric structures, in their each monomer, there exist two types of Cu centers that play pivotal roles as the components of electron transfer pathway in the process of catalysis. Furthermore, some residues alteration of Cu-NiRs would contribute to the catalytic reaction. In this review, the latest progresses about the construction features, the process of electron transfer and catalytic mechanism of Cu-NiRs were discussed.

Objective To evaluate combined buccal mucosa and lingual mucosa grafts for urethroplasty in a dog model. Methods Seven female mongrel dogs were selected.After a segment of proximal urethra mucosa (4 cm×1 cm) was excised and onlayed,urethroplasty was performed by using the combined free buccal mucosa (2 cm×1 cm)graft which had been harvested from the inferior cheek and free lingual mucosa graft(2 cm×1 cm)harvested from the inferior lateral surface of the tongue.A 12 F urethral catheter was kept for 7 d.Retrograde urethrography was done and urethra diameter was calibrated with a 10 F catheter before animals were sacrificed at week 12.Then the grafted areas excised and evaluated grossly and histopathologically. Results All dogs survived during the procedure and there was no tongue or bueeal complications.One dog developed a severe urethral stricture at the proximal anastomosis site.The remaining 6 dogs voided spontaneously with no difficulty.Retrograde urethrography showed that no stricture or fistula formed.The combined buccal mucosa graft and lingual mucosa graft shortened from a mean (SD) of 4.00(0.15)to 3.75(0.23)cm (statistically.significant,P＜0.05).No stricture was found in the connection of the buccaI mucosa and lingual mucosa grafts.Histological examination showed that the combined buccal mucosa and lingual mucosa grafts were well-incorporated into the urethral walls and covered by a keratinized squamous epithelium.Neovascularization was evident beneath the grafts. Conclusion Combined buccal mucosa graft and lingual mucosa graft could be an option for urethral substitution.

Objective To investigate the feasibility of constructing tissue engineered corpora cavernosa smooth muscle by seeding human umbilical artery smooth muscle cells (HUASMCs) in acel-lular collagen matrices.Methods Acellular corporal collagen matrices (ACCM) were obtained from the penis of adult rabbits by a cell removal procedure.HUASMCs were isolated from human umbilical cords through explant techniques and cultured in vitro.Subsequently,HUASMCs were seeded to ACCM and cultured in vitro.After that,the seeded ACCMs were implanted subcutaneously in 9 BALB/C athymic mice.Animals were killed 10,20 and 40 days after implantation.The implants were retrieved and morphological examinations were performed to evaluate characteristics of the engineered tissues.Additionally,organ bath studies were performed to address the contractility of the engineered tissues.Results The deeellularization process successfully extracted all cellular components; colla-gen fibers maintained their original porous morphology and structure.ACCM could be reseeded with cultured HUASMCs in vitro,and HUASMCs had the potential of attachment and proliferation on the three-dimensional ACCM scaffolds.Histologic analyses of the explants from all time points demon-strated a progressive regeneration of corpus cavernosum smooth muscle,with structures very similar to those of the native corpus cavernosum,The maximum contraction force induced by phenylephrine and electrical stimulation was (3.64+0.18)g and (2.50+0.21)g.Conclusion HUASMCs can be seeded on 3-dimensional ACCM scaffolds and will develop a tissue similar to that of the native corpus eavernosum smooth muscle.