Objective To build artificial dermis by using the acellular dermis,collagen membrane and collagen gel as scaffolds.Methods The fibroblasts were isolated from infant skin.The 3~(rd) generation cells were seeded into 3 different scaffolds.The artificial dermis was detected by HE staining,phase contrast microscope or scanning electron microscope.Results The fibroblasts implanted on the ADM began to rupture and died after 2 to 3 days.Though the fibroblasts proliferated well in collagen gel,the artificial dermis contracted obviously.Another artificial dermis contracted slightly by inoculating fibroblasts on collagen membrane,and the fibroblasts on them were in appropriate proliferation.Conclusion The artificial dermis built by collagen membrane as scaffolds has a preferable structure for an ideal substitute of skin.

Objective: To construct HBV adr gene vaccine pCMVS 2-S and to observe the specific humoral immune responses in C57BL/6 mice after gene immunization. Methods: The HBV gene vaccine was injected into tibialis anterior muscles of C57BL/6 mice. ELISA was used to detected the anti-HBs antibody in mice sera at various time points after gene transfer. Results: Anti-HBs in sera of mice was tested positive 1 week after plasmid injected, the level of antibody peaked 4 weeks later, and kept high titer for at least 2 months. Conclusion: The good response of humoral immunity can be induced by injection of pCMVS 2-S in C57BL/6 mice.

Objective To investigate the effect of sasanquasaponin (SQS) on injury of endothelial cells induced by hypoxia-reoxygenation and neutrophil adhesion, and its possible mechanisms. Methods Human umbilical vein endothelial cells (HUVEC) were exposed to normoxia or hypoxia-reoxygenation in the absence or presence of SQS (10.0, 1.0, and 0.1 ?mol/L). Lactate dehydrogenase (LDH) activity was determined in cultured HUVEC supernatants. HUVEC survival rate, neutrophil adhesion rate, malondialdehyde (MDA) level superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were measured. Neutrophil adhesion was assayed in additional HUVEC treated as above. Results The results showed that hypoxia-reoxygenation resulted in HUVEC injury and enhancement of neutrophil adhesion, with the increase in LDH activity, MDA level, and adhesion rate as well, conversely, with the decrease in activity of SOD and GSH-Px; SQS antagonized these changes in a concentration-dependent manner. Conclusion SQS may protect HUVEC against hypoxia-reoxygenation injury and its mechanism appeares to be related to anti-lipoperoxidization and anti-adhesion of white blood cell.

Objective To investigate the expression and effects of endothelin-1 (ET-1) in cerebral vessels of scalded rats. MethodHistological method was used to observe the alteration of histological structure of basilar artery in 8 scalded rats. Reverse transcription polymerase chain reaction(RT-PCR) was performed to examine expression of ET-1 mRNA;Western blotting to analyse expression of endothelin-1;radioimmunoassay to measure ET-1 levels;R3espectively, of cerebral basilar artery in 48 scalded rats. ResultVascular pathohistological changes were observed in cerebral basilar artery after rats were scalded;ET-1, ET-1 mRNA expression and ET-1 levels of basilar artery in scald 6 hours, 12 hours and 24 hours group were obviously higher than that of control group. Conclusion Scald may cause an increases of ET-1 expression in basilar artery, and elevated ET-1 expression may relate to the pathogenesis of cerebral vascular injury in scalded rats.

Objective To investigate the role of recombinant human interleukin 6 (rhlL-6) in calcification and osteogenic transition of cultured human umbilical artery smooth muscle cells (HUASMC), and the possible cell signal transduction way. Methods HUASMCs were isolated by the explant method. HUASMCs were treated with (treatment groups) or without (control group) rhIL-6. Alizarin Red S stain was applied for calcium deposition in extracellular matrix of control ceils and the cells treated with rhIL-6 50 μg/L at day 12. Calcium concentration in cell layer of control group and treatment group (treated with rhIL-6 10 μg/L and 50 μg/L, respectively) was determined calorimetrically by the o-cresolphthalein complexone method at day 3, 6, 9 and 12, and corrected by total cell proteins. The mRNA expressions of bone-specific alkaline phosphatase (BAP), osteopontin (OPN), bone morphogenetic protein-2 (BMP2) and osteoprotegerin (OPG) were estimated by real-time PCR in 12, 24 and 72 hours. OPN, BMP2 and OPG expressions were assessed by Western blotting and the BAP concentration at the same time was checked by fluorometry method . Electrophoretie mobility shift assays (EMSA) was used to detect the binding activity of transcription factor Cbfα1 with or without inhibitors of p38-MAPK (SB203580) and PKC (DHC) after 6 hours stimulation by rhIL-6 10 μg/L. Results rhIL-6 induced a positive Alizarin Red S stain and a time-dose-dependent increasing of cell layer calcium deposition.Compared with control group, rhIL-6 10 μg/L enhanced gene expression and protein levels of BAP and BMP2 at the early time (12 and 24 hours), and of OPN and OPG at later hours (24 and 72 hours). RhIL-6 still induced an increasing of binding activity of Cbfα1, which could be partially blocked by DHC but not SB203580. Conclusions rhIL-6 induces HUASMCs calcification and osteogenie transition in vitro, which may be one of the mechanism involved in IL-6 associated vascular calcification as observed in clinical studies. The role of IL-6 in HUASMCs may partially achieved through the PKC cell signal transduction way.

Objective To investigate the prevalence of glomerular microthrombosis in lupus nephritis (LN) and the significance of antibodies to anti-coagulation related factors and anti-phospholipid antibodies in glomerular microthrombosis (GMT).Methods Kidney biopsy specimens and plasma samples were obtained consecutively from 124 patients with LN. Kidney biopsy specimens were examined for the presence of glomerular microthrombi.Plasma samples from 25 LN patients with GMT (LN-GMT group) and 99 LN patients without GMT (LN-non-GMT group) were tested for lupus anticoagnlant (LA) and antibodies to cardiolipin (ACL),β2 glycoprotein I (β2GP I ),plasmin,thrombin,tissue plasminogen activator (t-PA) and Annexin A II.Results The prevalence of GMT in LN patients was about 20.2%.Compared to LN-non-GMT group,LN-GMT group had elevated SLE disease activity indices (SLEDAI),elevated activity and chronicity indices of kidney tissue injury,and elevated serum creatinine,blood urea nitrogen and proteinuria levels,and also had a higher frequency of hypertension (P＜0.01).The positive rates of LA,IgG class anti-β2GP I and anti-thrombin antibodies were higher in LN-GMT group than in LN-non-GMT group (P＜0.05).The positive rates of IgG class antibodies to ACL,plasmin,t-PA and Annexin A II in LN-GMT group were not statistically different from those in LN-non-GMT group (P＞0.05).No difference was found in the positive rate of any IgM class antibody between the two groups (P＞0.05).Conclusion This study has shown that GMT occurs approximately in 20.2% of the LN patients.Patients with GMT have more severe kidney tissue injury and more poor renal outcomes than patients without GMT.LA and antibodies to β2GP I and thrombin play a role in glomerular microthrombosis in lupus nephritis.

OBJECTIVE: To study the expression of angiopoietin receptor Tie-2 in the renal tissue of diabetic rats and the effects of Astragalus. METHODS: SD rats were randomly divided into normal control group, diabetes group and Astragalus-treated group. The expression of receptor Tie-2 in the renal tissue was assessed by using real-time quantitative polymerase chain reaction and immunohistochemical method. RESULTS: Glomerule Tie-2 protein expression was significantly elevated in the diabetes group as compared with the normal control group (P<0.01). Glomerule Tie-2 protein expression in the Astragalus-treated group was decreased as compared with the diabetes group (P<0.01). CONCLUSION: Tie-2 may play an important role in the pathogenesis of the early stage diabetic renal injury. The reno-protection effect of Astragalus may be mediated by down-regulating the expression of Tie-2 in the kidney tissue of diabetic rats.

Objective:To analyze the clinical manifestations and the diagnosis of the facial nerve tumor according to the clinical information, and evaluate the different surgical approaches depending on tumor location.Method:Twenty-seven cases of facial nerve tumors with general clinical informations available from 1999.9 to 2006.12 in the Shanghai EENT Hospital were reviewed retrospectively.Result:Twenty(74.1%) schwannomas,4 (14.8%)neuofibromas ,and 3(11.1%)hemangiomas were identified with histopathology postoperatively.During the course of the disease,23 patients(85.2%)suffered facial paralysis,both hearing loss and tinnitus affected 11 (40.7%)cases,5(18.5%)manifested infra-auricular mass and the others showed some of otalgia or vertigo or ear fullness or facial numbness/twitched. CT or/and MRI results in 24 cases indicated that the tumors originated from the facial nerve.Intra-operative findings showed that 24(88.9%)cases involved no less than 2 segments of the facial nerve,of these 24 cases 87.5%(21/24)involved the mastoid protion,70.8%(17/24)involved the tympanic protion, 62.5%(15/24)involved the geniculate ganglion, only 4.2%(1/24)involved the internal acoustic canal(IAC),and 3 cases (11.1%)had only one segments involved. In all of these 27 cases ,the tumors were completely excised,of which 13 were resected followed by an immediate facial nerve reconstruction,including 11 sural nerve cable graft,1 facial nerve end-to-end anastomosis and 1 hypoglossal-facial nerve end-to-end anastomosis.Tumors were removed with preservation of facial nerve continuity in 2 cases.Conclusion:Facial nerve tumor is a rare and benign lesion,and has numerous clinical manifestations.CT and MRI can help surgeons to make a right diagnosis preoperatively.When and how to give the patients an operation depends on the patients individually.

Objective To investigate the markers in early diagnosis of acute kidney injury (AKI) in patients undergoing heart surgery.Methods Markers included serum cystatin C (CyC),and urinary neutrophil gelatinase-associated lipocalin(NGAL),interleukin 18(IL-18),retinol binding protein(RBP)and N-acetyl-β-D-glucosaminidase(NAG).Twenty-nine cardiac surgical patients hospitalized were enrolled in the study.Serial blood and urine samples were collected immediately before incision and at various time intervals after surgery.The primary outcome measure was AKI.defined as a 50%increase in Scr from baseline. Results The cohort consisted of 29 patients aged(62.9±13.7)years,and baseline Scr was(73.2±11.9)μmol/L.There were no significant differences in demographics between cases and controls,while the aortic clamp time was predictably longer in AKI cases as compared to controls[(60.63±13.92)vs(43.00±9.20)rain,P<0.05].Each biomarker difiered significantly between cases and controls at least one timepoint.Optimal AUCs were for CyC at 10 houm with sensitivity (ST)0.71,specificity(SP)0.92,AUC=0.83(0.67-1.00),cut-off(CO)1.31 mg/L;NGAL at 0 hour with ST 0.84,SP 0.80,Auc=0.85(0.70-1.00),CO 49.15 μg/g Ucr;IL-18 at 2 hours with ST 0.85,SP 0.73,AUC=0.81(0.64-0.97),CO 285.65 ng/g Ucr;RBP at 0 hour with ST 0.75,SP 0.67,AUC=0.77(0.60-0.95),CO 2934.65μg/g Ucr and NAG at 4 hours with ST 0.86,SP 0.67,AUC=0.72(0.53-O.92),CO 37.05 U/mg Ucr.Using a combination of all the 5 biomarkers analyzed at the optimal time-point as above,an AUC of 0.98(0.93-1.02)(P<0.01)in this limited sample was able to obtain. Conclusions Application of serum and urinary biomarkers for the prediction of AKI in patients undergoing cardiac surgery is highly dependent on the sampling time.Of the evaluated markers,uNGAL has the best predictive profile.uRBP also shows similar predictive power.Combining all the five above biomarkers is able to predict significantly more cases,suggesting that the use of more than one marker may be beneficial clinically.

Objective To prospectively investigate the value of urinary neutrophil gelatinase-associated lipocalin (NGAL) in the diagnosis of acute kidney injury (AKI) following adult cardiac surgery. Methods Twenty-nine hospitalization patients undergone cardiac surgery were enrolled in the study. Serial blood and urinary samples were collected immediately before incision and at various time intervals after surgery. The primary outcome measure was acute kidney injury, defined as a 50% increase in the level of serum creatinine (Scr) from baseline. Results Fourteen of 29 developed acute kidney injury. The diagnosis time point with Ser was at 24 (10, 48) h after cardiac surgery. By contrast, the concentration of urinary NGAL rose from a median of 3.42(1.60, 9.92) μg/L at baseline to 20.51(13.42, 50.02) μg/L at admission to ICU (P=0.006),and the median concentration of urinary NGAL in patients who developed AKI was significantly higher at admission to ICU compared with patients who did not develop AKI [20.51 (13.42, 50.02) μg/L vs 2.91 (0.72,8.61) μg/L, P=0.002]. As to urinary NGAL at admission to ICU, the area under the receiver-operating characteristic (ROC) curve was 0.824, the sensitivity was 85.7% and the specificity was 80.0% with a cutoff value of 10.95 μg/L. Significant correlation was found between urinary NGAL at admission and the levd of Scr at 24 h in ICU (r=0.545, P=0.002), as well as estimated GFR (r=-0.546,P=0.002). Conclusion Urinary NGAL concentration is significantly higher in patients developing postoperative AKI at the early time of admission to ICU, which may be a useful early biomarker of AKI after adult cardiac surgery.