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 determine the triterpenoid content in Loquat leaf by using HPLC method and to explore the feasibility of ursolic acid and oleanolic acid as the quantitative markers for determination of Loquat leaf and its preparations.Methods Two desugaring methods of Loquat leaf extracts were compared.The contents of ursolic acid and oleanolic acid in Loquat leaf as well as preparations were measured based on a system of HPLC,with a Dumas Diamonsil C18 column (250 mm×4.6 mm,5 μm) was used as analytical column.The mobile phase was consisted of acetonitrile-0.1% phosphoric acid (95∶5) with the flow rate of 0.5 mL·min-1.The column temperature was 30 ℃,the injection volume was 10 μL, the detective wavelength was 210 nm.Results Ursolic acid and oleanolic acid were measured in both Loquat leaf and its preparations,and their contents were fairy stable and comparable.Conclusion Ursolic acid and oleanolic acid can be served as quantitative markers for the determination of Loquat leaf and its preparations.

Objective To measure the renal tissue texture or flexibility with virtual touch quantization (VTQ) and to tentatively examine its clinical application in patients with chronic kidney disease(CKD).Methods A total of 750 patients (1500 kidneys) were performed with VTQ,including 400 cases in the control group,and 350 cases in the CKD group.A conventional ultrasound examination (two-dimensional,color Doppler) were first taken,and then the shear wave velocity (Vs) was measured which reflected the textural elastic.Results In both groups the Vs was the highest in renal cortex with significant difference (P<0.05); renal cortical region Vs in CKD group was lower than those in control group (P<0.05),while Vs of renal medulla and renal sinus had no significant difference in the two groups.The severity of renal dysfunction was increased along with a Vs decrease of renal cortex.Conclusion VTQ is helpful to assess renal function of patients with CKD.

AIM:To observe the efficacy of intravitreal conbercept injection for chronic central serous chorioretinopathy (CSC).METHODS: Nine eyes of 9 patients diagnosed as chronic CSC between October 2015 to May 2016 were treated with an intravitreal injection of conbercept (0.5mg/0.05mL) (six patients were given the same does of intravitreal injection again at 1mo after the first injection).Follow-up observation was at 1, 2, and 6mo after injection.Observed indicators included best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal indocyanine green angiography (ICGA), macular fovea thickness (CMT), subfoveal choroidal thickness (SFCT).RESULTS:Seven of the 9 patients responded significantly to the drug, while 2 patients had no response.The CMT was 373.12±72.43μm at baseline, which decreased significantly to 332.05±67.13μm, 282.24±62.30μm and 225.56±71.08μm at 1, 2 and 6mo after the intravitreal injection.The mean thickness of SFCT was 422.11±64.82μm before treatment.The choroidal thickness of non-responsive patients before treatment was below average, respectively 353μm and 365μm.The SFCT of 1, 2, and 6mo after treatment was 391.45±75.24μm, 365.53±63.07μm, 355.40±66.65μm.Before treatment and 1mo after, there was no significant difference (P=0.074), but there was statistically significant (P<0.01) between those of before and 2mo and 6mo after.The mean BCVA of the prior treatment was 0.53±0.32, the after treatment was 0.65±0.20, there was no different between the two(P>0.05).CONCLUSION: Intravitreal conbercept injection in chronic CSC may have some effect in accelerating subertinal fluid resolution and decreasing the CMT.The SFCT within 6mo after treatment was significantly lower than pretreatment.The SFCT may be an indicator of whether patients respond.

Poly(ADP-ribose) polymerase-1 (PARP-1) has emerged as a promising anticancer drug target due to its key role in the DNA repair process. It can polymerize ADP-ribose units on its substrate proteins which are involved in the regulation of DNA repair. In this work, a novel series of para-substituted 1-benzyl-quinazoline-2, 4 (1H, 3H)-diones was designed and synthesized, and the inhibitory activities against PARP-1 of compounds 7a-7e, 8a-8f, 9a-9c and 10a-10c were evaluated. Of all the tested compounds, nine compounds displayed inhibitory activities with IC50 values ranging from 4.6 to 39.2 micromol x L(-1). In order to predict the binding modes of the potent molecules, molecular docking was performed using CDOCKER algorithm, and that will facilitate to further develop more potent PARP-1 inhibitors with a quinazolinedione scaffold.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Drug Design ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Molecular Docking Simulation ; Molecular Structure ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; Quinazolinones ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship

By means of preparative HPTLC and column chromatography over silica gel and Sephadex LH-20, ten sesquiterpenes were isolated and purified from the whole plants of Solanum septemlobum Bunge. Based on the physico-chemical properties and spectral data, their structures were elucidated and identified as: lyratol D(1), solajiangxin B(2), 1 ,2-dehydrocyperone(3), solanerianone A (4), dehydrocarissone(5), ligucyperonol(6), nardoeudesmol A(7), solajiangxin F(8), and lyratol B(9), solajiangxin D(10). For the first time, compounds 1-10 were isolated from Solanum septemlobum, and compounds 5-7 were obtained from the genus Solanum.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Sesquiterpenes ; chemistry ; isolation & purification ; Solanum ; chemistry

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

Epithelial-to-mesenchymal transition (EMT) plays a critical role in cancer metastasis,and is relevant to the inflammatory microenvironment.Lipopolysaccharide (LPS),a cell wall constituent of gram-negative bacteria,has been reported to induce EMT of cancer cells through TLR4 signal.We previously reported that LPS promoted metastasis of mesenchymallike breast cancer cells with high expression of cyclin D 1 b.However,the role of cyclin D1b in LPS-induced EMT has not been fully elucidated.In the present study,we described that cyclin D1b augmented EMT induced by LPS in MCF-7 breast cancer cells.Cyclin D1b markedly amplified integrin αvβ3 expression,which was further up-regulated under LPS stimulation.Our results showed ectopic expression of cyclin D1b promoted invasiveness of epithelial-like MCF-7 cells under LPS stimulation.Additionally,LPS-induced metastasis and EMT in MCF-7-D1b cells might depend on αvβ3 expression.Further exploration indicated that cyclin D1b cooperated with HoxD3,a transcription factor promoting αvβ3 expression,to promote LPS-induced EMT.Knockout of HoxD3 repressed LPS-induced EMT and αvβ3 over-expression in MCF-7 cells with high expression of cyclin D1b.Specifically,all these effects were in a cyclin D1a independent manner.Taken all together,LPS up-regulated integrin αvβ3 expression in MCF-7 cells with high expression of cyclin D 1b and induced EMT in breast cancer cells,which highlights that cyclin D1b may act as an endogenous pathway participating in exogenous signal inducing EMT in breast cancer cells.

Objective To explore the effect of upgrade of dialysate quality on the microinflammation in maintenance haemodialysis (MHD) patients. Methods Fifty-three MHD patients in Kidney Center of the First Affiliated Hospital,Medical College,Zhejiang University in January 2003 were enrolled in the prospectively study.The main end-points were survival at 8 years or weaning from haemodialysis during 8-year period including death, receiving renal transplantation or transferring to peritoneal dialysis.The endotoxin level of dialysate and patients' serum levels of interleukin-6 (IL-6),tumor necrosis factor α (TNF-α),C reaction protein (CRP),and albumin were recorded during the observation period. Results After the upgrade of water management system,endotoxin level of dialysate obviously decreased [(0.046±0.012) EU/ml vs (0.454±0.002) EU/ml,P＜0.01],and serum IL-6 [(3.947±3.624) ng/L vs (13.779±7.106) ng/L,P=0.036],TNF-α [(7.935±3.864) ng/L vs (12.804±8.017) ng/L,P=0.012] as well as CRP [(0.194±0.149) mg/L vs (0.561 ±0.309) mg/L,P＜0.01] decreased significantly,while serum albumin increased [(41.900±6.803) g/L vs (38.140±7.083) g/L,P=0.042].Hemoglobin level did not change significantly after the system upgrade,however,the dose of erythropoietin decreased [(93.0±12.7) U·kg-1·week-1 vs (131.0±10.1) U·kg-1·week-1,P=0.015]. Conclusions The upgrade of central dialysis fluid delivery and water management system by application of double reverse osmosis,high frequency heat disinfection and endotoxin filter can improve the quality of dialysate.Improvement of dialysate quality can ameliorate the microinflammation state of MHD patients.

Objective To investigate whether erythropoietin(Epo) is potentially beneficial in protecting cultured retinal neurocytes.Methods Primary isolated retinal nerve cells were cultured.Expressions of Epo and EpoR protein in cultured retinal neurocytes were decected by immunohistochemical analysis.Survival of cultured neurocytes that were incubated in the presence of Epo or glutamate in the presence or absence of Epo were estimated by determining the activity of their mitochondrial dehydrogenases using the MTT assay.Results Epo and EpoR protein were expressed on the cultured retinal neurocytes.The presence of different concentrations of Epo did not improve the survival of retinal neurocytes,and Epo could prevent glutamateinduced toxicity. Conclusion Epo is beneficial in protecting mixed cultured retinal neurocytes from glutamate-induced cytotoxicity.