Objective To evaluate the combined detection of serum galactomannan (GM test) and 1,3-β-D-glucan (G test) in diagnosis of invasive pulmonary aspergillosis (IPA).Methods A total of 136 patients suspected for IPA were enrolled in the study from Jinhua Central Hospital in Zhejiang Province from January 2007 to December 2011.GM and G tests were performed.x2 test was used to conpare the sensitivity,specificity,positive predictive value and negative predictive value of individual test and combined test,and the area under the ROC curve (AUC) was used to determine the effectiveness of the tests.Results The sensitivity and specificity of G test were 84.0％ and 80.9％,and those for GM test were 78.0％ and 88.2％,respectively.The combination of two tests (parallel) increased sensitivity to 92.0％ and the serial test increased the specificity to 92.6％.The AUC of the combined test was 0.923(95％ CI:0.867-0.980).Conclusion The combination of GM test and G test can improve the diagnostic effectiveness for IPA.

ObjectiveTo investigate the value of cavity isolation method on left colorectal obstruction in the anastomosis isolation. Methods60 patients with left colorectal obstruction were randomly divided into two groups:ileostomy group and cavity isolation method group,30 patients for each. After left colon resection was completed, ileostomy anastomosis group used the traditional method for colorectal anastomosis, and protected the anastomosis by ileostomy. Cavity isolation method group protected the anastomosis by cavity isolation method. Then the indexes in the two groups,such as anastomotic height, operation time, anal exhaust time,incidence of anastomotic leakage and its complications,hospitalization time, medical cost and so on were analyzed. Results Longer postoperative hospital stay (9.3 + 4.2) d were observed in ileostomy group than those ( 15.6 + 3.3 ) d ( P ＜ 0.05 ). Earlier time of the first flatusand oral intake(87.2 ± 6.3 ) h, ( 91.2 ± 7.2 ) h were observed in ileostomy group than those ( 101.8 + 7.5 ) h,( 117.3 ± 8.6) h in cavity isolation method group ( all P ＜ 0.05 ). ConclusionThe anastomosis isolation was a safe and feasible new technology that could reduce the complications caused by anastomotic leakage, and it had the advantages of short hospital stay, low cost,less suffering ect.

Objective To explore the mechanism and the therapeutic effects of the pelvic floor muscle electrical stimulation on female patients with urinary incontinence Methods Forty three female patients with incontinence were recruited. With different electrical currents, the pudendal nerves were intermittently stimulated by the electrode probe placed in the middle of vagina. To prevent incontinence,the contractions of the detrusor were depressed and the capacity of the bladder was increased by neural reflex, while the urethra obstruction was enhanced by the contractions of pelvic floor muscles. Results The incidence of urethrorrhea was decreased by 46%, and the frequency of micturition was reduced by 49%, while the frequency of urination in 24h ranged from 10 to 12. The general subjectively rated improvement rate of patients was 68%, while the general objectively one was 74%. Conclusion The pelvic floor muscle electrical stimulation, which can improve the patients' life quality, has significant therapeutic effects on female patients with incontinence.

Objective To investigate the possible mechanism that hepatocyte growth factor (HGF) inhibits renal tubular epithelial-mesenchymal transition (EMT),and to determine whether Smurf2 expression induced by TGF-β1 can be reversed by HGF in normal rat kidney epithelial cells (NRK-52E).Methods Using rat NRK-52E cell line as an in vitro system,NRK-52E cells were incubated with 5 μg/L TGF-β1 for 0-24 h.Part of cells were pretreated with 20 μg/L HGF for 30 min or not,then incubated with or without 5 μg/L TGF-β1 for 1 h or 48 h.The other cells were transfected with pFlag-Smurf2 or Smurf2 siRNA for 24 h,then treated with or without 20 μg/L HGF for 24 h.The expressions of Smurf2,SnoN,E-cadherin,alpha-smooth muscle actin (α-SMA) and fibronectin (FN) were detected by Western blotting and indirect immunofluorescence staining assays.Results Compared to normal control,TGF-β1 could rapidly induce Smurf2 protein expression in a short time (P＜0.01).Meanwhile,the expressions of FN and α-SMA were significantly induced,and the expression of E-cadherin was reduced in NRK-52E cells by TGF-β1.In contrast,in the NRK-52E cells pretreated with HGF,HGF could obviously inhibit Smurf2 expression induced by TGF-β1,and reversed the down-regulation of SnoN (P＜0.01) and E-cadherin (P＜0.05),the up-regulation of α-SMA (P＜0.01) and FN (P＜0.01) induced by TGF-β1.Moreover,overexpression of Smurf2 in NRK-52E cells could partly inhibit the up-regulation of SnoN protein by HGF,while down-regulation of Smurf2 could up-regulate the expression of SnoN induced by HGF.Conclusions HGF can abolish EMT induced by TGF-β1 in renal tubular epithelial cells through down-regulating Smurf2 expression and suppressing ubiquitin-proteasome dependent degradation of SnoN.

Objective To investigate the relationship between high-glucose-induced fibronectin(FN) expression and up-regulation of integrin-linked kinase(ILK) in human kidney tubular epithelial cells (HKC) and kidney of CD-1 mice. Methods Cultured human kidney tubular epithelial cells and streptozotocin (STZ)-indueed diabetic model of CD-1 mice were enrolled in this study.Western blot was used to detect the expression of FN and ILK.The kinase dead ILK plasmid (pCMV-kdlLK) were transferred to HKC. Results Four weeks after injection of STZ,CD-1 mice had higher blood glucose level as compared to the control [(20.3±2.7) mmol/L vs (6.1±1.4) mmol/L,P＜0.01].Meanwhile,expression of FN and ILK was significantly increased in diabetic mice as compared to the control (P＜0.01).There was positive correlation between the expression of FN and ILK (r=0.899,P＜0.01).High-glucose could up-regulate FN and ILK expression in cultured HKC in a time- and dose-dependent manner.Blockage of ILK activation by pCMV-kdILK abrogated high-glucose-incuced FN expression in HKC. Conclusions Highglucose can induce FN expression through up-regulating ILK expression.Blockage of ILK activation abrogates this effect.

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective To investigate the effect of high glucose on renal tubular epithelial-mesenchymal transition,and to analyze the relationship between high glucose and transforming growth factor β1(TGF-β1)and the mechanism of renal interstitial fibrosis. Methods HKC and Smad7-overexpression HKC cells were grown in DMEM/F12 medium containing 5%～10%newborn calf serum.They were cultured for 16 h in free serum medium after 80%cells were adhered onto the surface of the flask.Afterwards,they were stimulated by high glucose(glucose concentration:25 mmol/L and 50 mmol/L).The expression of α-SMA,E-cadherin and fibronectin was detected by Western blot while the supernatant level of TGF-β1 was detected by ELISA.Cell motility and migration was evaluated using Boyden chamber motogenicity assay. Results In HKC induced by high glucose,the expression of α-SMA and fibronectin protein was highly upregulated while the expression of E-cadhefin protein was down-regulated.The expression of TGF-β1was up-regulated in a dose-dependent manner.These above-mentioned effects could be obviously inhibited by anti-TGF-β1 antibody.The protein expression of α-SMA,fibronectin and E-cadherin had no obvious change in Smad7-overexpression HKC induced by high glucose.HKC exhibited enhanced motility and invasive capacity in high glucose groups,compared to that in control group.Migrated cell counting was(12.4±3.7)and(18.6±4.4)cell/HP in 25 and 50 mmol/L glucose groups respectively. Conclusion High glucose may induce renal tubular epithelialmesenchymal transition through TGF-β1 pathway,which can be inhibited by blocking the Smad signal pathway.

Objective To evaluate the immune effects of virus-like particles ( VLPs) of VP1 pro-teins derived from norovirus GⅠ. 1 and GⅡ. 4 genotypes expressed in Hansenula polymorpha expression sys-tem. Methods SDS-PAGE and Western blot assay were performed to detect the purity of GⅠ. 1 and GⅡ. 4 VP1 proteins after purification. Morphologies of the recombinant VLPs were observed under transmission electron microscopy ( TEM) . Sizes and distributions of the VLPs were analyzed by dynamic light scattering analyzer. BT50(50% of blocking titer) was detected by HBGA (histo-blood group antigen) blocking assay in BALB/c mice immunized with different regimens. Results SDS-PAGE analysis of the purified recombinant GⅠ. 1 and GⅡ. 4 VP1 proteins showed that their purity were greater than 90%. Western blot assay con-firmed the specific bands of VLPs. TEM images showed that the sizes of purified GⅠ. 1 and GⅡ. 4 VP1 VLPs were at a mean diameter of 30-50 nm with clear border and high homogeneity, which was similar to that of wild virus. BT50 significantly increased in the groups, in which Al( OH) 3 was used as adjuvant. Con-clusion Animal studies have shown that administration of GⅠ. 1 and GⅡ. 4 VP1 VLPs in the presence of Al( OH) 3 induces detectable HBGA-blocking antibody, indicating that GⅠ. 1 and GⅡ. 4 VP1 VLPs are promising candidates for norovirus vaccine.

AIM: To investigate the effect of propofol on expression of protein kinase C (PKC) mRNA during pulmonary ischemia and reperfusion injury (PIRI) in rabbits. METHODS: Single lung ischemia and reperfusion animal model was used in vivo. The rabbits were randomly divided into three groups ( n =9 in each): sham operated group (sham), PIR group (I-R) and PIR+ propofol group (PPF). Changes of several parameters including malondialdehyde (MDA), superoxide dismutase (SOD), wet to dry ratio of lung tissue weight (W/D) and index of quantitative assessment of histologic lung injury (IQA) were measured at 60 minutes after reperfusion in lung tissue. Meanwhile the location and expression of PKC mRNA were observed. Lung tissue was also prepared for light microscopic and electron microscopic observation at 60 minutes after reperfusion. RESULTS: As compared with group I-R, PKC mRNA strongly expressed in intima and extima of small pulmonary artery as well as thin-wall vessels (mostly small pulmonary veins) in PPF group. The average optical density values of PKC-?、? and ? mRNA in small pulmonary veins PPF in group showed significantly higher than that in I-R group (all P

Objective To summary the experience of surgical types selection of chronic suppurative otitis media tympano‐plasty and evaluate its clinical curative effect .Methods Chronic suppurative otitis media patient records were collected ,surgical types selection based on hearing ,otoscope and mastoid CT selection .Finally 161 chronic suppurative otitis media patients (164 ears) whoes medical records and follow‐up of contents were complete were collected ,in which type Ⅰ tympanoplasty group with 40 ears , intact bridge radical mastoidectomy+ type Ⅰ tympanoplasty group with 30 ears ,and open radical mastoidectomy+type Ⅱ tympan‐oplasty group with 94 ears .The therapeutic effects of the three surgical methods for the treatment of chronic suppurative otitis media were evaluated from hearing ,quality of life as well as satisfactory score after surgery .Results The mean values of the air conductive hearing thresholds after operation in the three tympanoplasty groups were decreased than those before operation (all P<0 .05) ,while the mean values of the bone conductive hearing thresholds after operation showed no obvious changes (all P>0 .05) .There were no statistically significant improvements in the air conductive hearing thresholds in the three tympanoplasty groups (P>0 .05) .The improvements in patient′s subjective hearing of the there surgical methods were 82 .5% ,70 .0% and 83 .0%respectively ,there was no statistically significant (P>0 .05) .The improvements in the overall quality of life of the three surgical methods were 80 .0% ,80 .0% and 79 .8% respectively ,showed no statistically significant differences (P>0 .05);the reduction rate of patient′s worry about illness were 90 .0% ,93 .3% and 95 .7% ,showed no statistically significant differences (P>0 .05);the bas‐ic satisfaction rates in the postoperative satisfaction ratings of the three surgical methods were 100 .0% ,100 .0% and 97 .9% ,and there were no statistically significant differences (P>0 .05) .Conclusion For tympanoplasty in chronic suppurative otitis media pa‐tients ,surgical types selection based on hearing ,otoscope ,mastoid CT selection and intraoperative findings .From the evaluation of postoperative patients with hearing and quality of life ,three surgical methods of chronic suppurative otitis media Tympanoplasty a‐chieve satisfactory results .