Human kerotinocyte growth factor (hKGF) gene amplified by PCR was inserted into the shuttle vector pAdTrack-CMV to get the recombinant plasmid pAdTrack-CMV-hKGF, which was linearized with Pme I and transferred into Escherichia coli BJ5183 containing the adenoviral bone plasmid pAdEasy-1 to obtain the recombinant adenoviral plasmid pAdEasy-hKGF. The recombinant adenoviral plasmid was then transfected into HEK-293 cell lines via Lipofectamine 2000 to package and amplify the recombinant adenovirus containing hKGF gene detected by PCR. The recombinant adenovirus produced could effectively infect HaCat cells. The result of Western blotting showed that HaCat cells infected with the recombinant adenovirus expressed and secreted hKGF protein.

Objective To investigate the chemical constituents of the Radix Campylotropidis Hirtellae ( RCH ) . Methods 60%ethanol extract of the RCH was separated by column chromatography on silica gel, ODS and Sephadex LH-20 to obtain aimed compounds.Structures of the compounds were investigated by modern spectroscopy.Results Nine compounds were separated from Campylotropis hirtella ( Franch.) Schindl, and the structure of the new compound 1 was determined by spectral analysis and named hedysarimcoumestanⅠ( 1 ) , while the others were identified as the known isotrifoliol(2), hedysarimcoumestan B(3), umbelliferone(4), orobol(5), quercetin(6),(+) catechin(7), kaempferol (8) and dihydrokaempferol(9) .Conclusion Compound 1 is a new compound and 3-5 are separated from C.hirtella for the first time.The screening results of inhibition of H5N1 virus activity show that compound 5 has some inhibitory activity at a level of 50.0 μg /ml.

Objective To investigate the clinical characteristics and treatments of endometrioid carcinoma patients with preoperative diagnosis of endometrial hyperplasia.Methods From 2005 to 2010, 404 patients were diagnosed with endometrioid carcinoma after hysterectomy.Among these patients,44 of them were diagnosed atypical endometrial hyperplasia(AEH) preoperatively.Retrospectively analysis the characteristics of these patients with SPSS13.0.Results Among the 44 cases, all of them were grade G1 disease, and 39 of them received comprehensive staging surgery.14(32%)young cases preserved bilateral ovaries.9 cases(20%) were given adjuvant radiology.No recurrence was detected during the median follow up of 52 months.Compared to the premenstrual group, although no statistical difference was detected, more patients with risk factors of deep myometrium invasion(4/22 vs 1/22) and lymph-vascular space invasion(LVSI, 3/22 vs 0/22) in the postmenstrual group.Compared to the patients who diagnosed with endometrioid cancer(EC) preoperatively, there are more patients with grade G1(P=0.000), fewer patients received adjuvant chemotherapy(P=0.003) and fewer recurrence(P=0.019) in AEH group.Conclusions The endomtrioid cancer patients who diagnosed with atypical hyperplasia preoperatively have better prognosis.Hysterectomy with bilateral ovaries preserved is acceptable in young patients.Post menopause patients have more risk factors of deep myometrium invasion and LVSI.

ObjectiveTo evaluate the effect of rehabilitation nursing in the peri-operative period of the first-stage combined anterior-posterior approach surgery in the sacral tumor resection.Methods15 patients with sacral tumor received psychological intervention, gastrointestinal preparation, exercises of perineum sphincter contraction and relaxation training before the operation. Changes of patients' vital signs and nerve function were observed after the operation, and rehabilitation training and instruction were performed.ResultsAll cases were followed up for 2～37 months. Except one case died of tumor transfered to lung, all of other cases had a satisfactory curative effect in the near future, and their local symptom released and the functions of urination and defecation were kept well.ConclusionRehabilitation nursing in the peri-operative period plays an important role in surgical treatment of sacral tumor. It affects the rehabilitation and function restitution obviously and can reduce the occurrence of complication effectively.

Adenomatous Polyposis Coli Protein ; genetics ; Chromatography, High Pressure Liquid ; methods ; Codon ; genetics ; DNA Mutational Analysis ; Fibromatosis, Aggressive ; genetics ; pathology ; Humans ; Mutation ; Polymerase Chain Reaction ; beta Catenin ; genetics

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.
Adenoviridae ; genetics ; Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Fibroblast Growth Factor 10 ; genetics ; metabolism ; Humans ; Keratinocytes ; cytology ; metabolism ; Proteomics ; methods ; RNA, Messenger ; metabolism ; Recombinant Proteins ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Transfection ; Up-Regulation ; Voltage-Dependent Anion Channel 2 ; biosynthesis ; metabolism

To study the role of anti- C. albicans IgY and serum in protection of C. albicans infection of several animal models. To develop three animal models of C. albicans infection: a burned rat model of C. albicans infection, a mouse model of vaginal candiasis and a immunosuppression mouse model of C. albicans infection. And we compared the contribution of anti- C. albicans IgY and serum to the clearance of the C. albicans in three animal models of C. albicans infection. Anti- C. albicans IgY can protect against C. albicans infection in a burned rat model of C. albicans infection and a mouse model of vaginal candidiasis. The serum can effectively protect the mice from disseminated candidiasis in a immunosuppression mouse model. Humoral immunity component involving anti- C. albicans IgY and serum protect against C. albicans infection in a burned rat model of C. albicans infection ,a mouse model of vaginal candidiasis and a immunosuppression mouse model of C. albicans infection.

Objective To compare clinical outcomes in eyes with macula-off rhegmatogenous retinal detachments (RRD) with peripheral breaks managed by surgical protocols that result in either complete (CSFD) or partial subretinal fluid drainage (PSFD).Methods Following the clinical detection of a maculaoff RRD with peripheral retinal breaks,patients were offered the opportunity to enroll in the study,and those patients who signed the consent were evaluated for eligibility based upon the inclusion and exclusion criteria for this clinical study,and if fully eligible they were assigned prospectively to one of the two surgical designs (PSFD or CSFD,1∶ 1) using a random number table.Seventy-two eyes of 72 patients were enrolled and studied.Patients were treated with 25G plus vitrectomy,endolaser or transscleral cryopexy,either complete (n=36),or partial (n=36) subretinal fluid drainage,and 14％C3F8 (PFO) was used for intraocular tamponade.After surgery,all patients were kept in a supine position for 24 hours,and then in a clinically optimal position for 6-10 days.The study patients were examined at 1,3 and 6 months after surgery with thorough ophthalmic examinations.Macular optical coherence tomography (OCT) imaging was acquired in 1 month.Anatomical and visual outcomes as well as intra-operative and postoperative complications of the two groups were compared.Furthermore,the persistence of subfoveal fluid in OCT images and the symptoms of distortion at 3 months were measured and recorded.The primary study endpoint of anatomic retinal reattachment for each group was based upon the 6-month time-point.Results The preoperative baseline characteristics between the two groups were not significantly different.The single-operation success rates were 88.9％ and 91.6％ respectively for the CSFD and the PSFD groups (x2 =0.158,P＞0.05).The mean best corrected visual acuity (BCVA) at 6 month endpoint were 0.99± 0.52 minimum resoluation angle in logarithmic (logMAR) for the CSFD group and 1.07±0.34 logMAR for the PSFD group(t=0.580,P=0.564).The mean operative time was longer in the CSFD group (62.25± 4.32) minutes than that in the PSFD group (47.9 ± 5.0) minutes (t =0.580,P=0.564).seven of 29 (24.1％) phakic eyes in the CSFD group had lens injury during SRF drainage,and none of the 31-phakic eyes in the PSFD group sustained lens damage.Residual PFO was present in 6 of 36 CSFD cases (16.7％).Successful retinal reattachment after primary surgery was achieved in 33) PSFD eyes and in 32 CSFD eyes based upon OCT imaging at 1 month demonstrated reattached foveae with no residual subfoveal fluid.Among these patients,22 patients (62.5％) in the CSFD group and 23(69.7％) patients in the PSFD group reported distortion in the operated eye or/and a difference in image size between the two eyes at the 6 month visit (P=1.00).Conclusions Partial subretinal fluid drainage during pars plana vitrectomy for the repair of macula-off RRD with peripheral breaks is effective.The success rates are not statistically different.Additionally,PSFD procedures can simplify the surgery procedure,shorten operative time and,and to some extent,reduce the incidence of complications relevant to the CSFD approach.

AIM: To investigate the effects of bone morphogenetic protein 7 ( BMP-7 ) on the expression of transcription factor E2A and inhibitor of differentiation 2 (Id2) in the renal tubule epithelial cells(NRK-52E)exposed to high glucose, and to explore its possible mechanism of improving renal tubular fibrosis induced by high glucose.METH-ODS:The NRK-52E cells were divided into control group, high glucose (HG) group and high glucose with different doses of BMP-7 (10μg/L and 20μg/L) group.The cells in HG group and BMP-7 group were cultured for 12 h, 24 h and 48 h. The protein expression of Id2, E2A, E-cadherin,α-smooth muscle actin (α-SMA) and collagen-I was detected by Western blot.In addition, the mRNA expression of Id2 was detected by real-time PCR.RESULTS:Compared with control group, the mRNA and protein levels of Id2 and the protein level of E-cadherin were down-regulated, while the protein levels of E2A,α-SMA and collagen-I were up-regulated in HG group (P<0.05).Compared with HG group, the mRNA and pro-tein levels of Id2 and the protein level of E-cadherin were significantly up-regulated, while the protein expression of E2A,α-SMA and collagen-I was significantly down-regulated in 20 μg/L BMP-7 group ( P<0.05 ) .The correlation analysis showed that the Id2 protein level was negatively correlated with the E2A protein level (P<0.05).CONCLUSION:BMP-7 may intercept the process of renal tubule fibrosis induced by high glucose via promoting the expression of Id2 and inhibi-ting the expression of E2A at protein level.

Objective To observe the clinical effect of intravitreal ranibizumab (IVR) combined with vitrectomy in treating proliferative diabetic retinopathy (PDR).Methods This is a prospective non-randomized controlled clinical study.A total of 62 patients (70 eyes) who underwent vitrectomy for PDR were enrolled and divided into IVR group (30 patients,34 eyes) and control group (32 patients,36 eyes).IVR group patients received an intravitreal injection of 0.05 ml ranibizumab solution (10 mg/ml) 3 or 5 days before surgery.The follow-up time was 3 to 18 months with an average of (4.5± 1.8) months.The surgical time,intraoperative bleeding,iatrogenic retinal breaks,use of silicone oil,the best corrected visual acuity (BCVA) and the incidence of postoperative complications were comparatively analyzed.Results The difference of mean surgical time (t=6.136) and the number of endodiathermy during vitrectomy (t=6.128) between IVR group and control group was statistically significant (P=0.000,0.036).The number of iatrogenic retinal break in IVR group is 8.8 ％ and control group is 27.8％,the difference was statistically significant (x2 =4.154,P=0.032).Use of silicone oil of IVR group is 14.7％ and control group is 38.9％,the difference was statistically significant (x2 =5.171,P=0.023).The incidence of postoperative vitreous hemorrhage in 3 month after surgery was 11.8％ and 30.6 ％ respectively in IVR group and control group.The differences were statistically significant (x2=3.932,P=0.047).The 6 month postoperative mean BCVA of IVR group and control group have all improved than their preoperative BCVA,the difference was statistically significant (t=4.414,8.234;P=0.000).But there was no difference between the mean postoperative BCVA of two groups (t=0.111,P=0.190).There was no topical and systemic adverse reactions associated with the drug after injection in IVR group.Conclusions Microincision vitreoretinal surgery assisted by IVR for PDR shorten surgical time,reduces the intraoperative bleeding and iatrogenic retinal breaks,reduces the use of silicon oil and the postoperative recurrent vitreous hemorrhage.But there was no significant relationship between vision improvement and IVR.