Objective To investigate the relationship between interleukin-17 (IL-17) and vitiligo.Methods Totally,32 vitiligo patients and 30 healthy human controls were enrolled in this study.Blood samples were collected from all the subjects,and enzyme-linked immunosorbent assay (ELISA) was performed to determine the plasma levels of IL-17.The relationship of plasma IL-17 levels with disease stage,clinical course and lesion area was assessed.Results The plasma levels of IL-17 were significantly higher in the patients with progressive and stable vitiligo than in the healthy controls (both P ＜ 0.05),and higher in the patients with progressive vitiligo than in those with stable vitiligo (P ＜ 0.05).Moreover,the plasma levels of IL-17 were positively correlated with the area of vitiligo lesions (r =0.456,P ＜ 0.05),but unrelated to the clinical course of vitiligo (r =0.239,P ＞ 0.05).Conclusion IL-17 may play a certain role in the occurrence and development of vitiligo.

Objective To observe the effects of the topical benzalkonium chloride on the cornea of experi-ment animals and evaluate the safety of topical benzalkonium chloride.Methods 36 rabbits were randomly divided into two groups according to the random number table,18 rabbits(36 eyes)in each group.The benzalkonium group was assigned as preservative benzalkonium chloride(0.05%)gutta,the control group was served as blank control. Rabbits were examined by the slit lamp,non-contact corneal endothelial microscope and Ultrasonic Wave instrument for corneal endothelial cell density and corneal thickness before and the 3rd weeks,the 6th weeks after the medica-tion.In vitro cornea was stained by typan blue and alizarin red,corneal endothelial cells were observed and calculate the viability of corneal endothelial cells (CECs).Results The differences of corneal thickness at the 3rd weeks and the 6th weeks were not statistically significant between the two groups(t =1.876,1.876,all P >0.05);The differ-ences of the corneal endothelial cell density and the viability of corneal endothelial cells(CECs)between the two groups at the 3rd weeks and the 6th weeks were not statistically significant(t =0.321,1.541,all P >0.05);The tear meniscus height of the benzalkonium group were lower than the control group at the 6th weeks[(142 ±36)μm vs. (198 ±31)μm,t =2.462,P <0.05].Conclusion Benzalkonium chloride has no impact on the corneal endothelial cells in short period,but the affection on the tear is obvious.

The known members of inhibitor of growth (ING) gene family are considered as candidate tumor suppressor genes. ING4, a novel member of ING family, is recently reported to regulate brain tumour angiogenesis through transcriptional repression of NF-?B-responsive genes, induce G2/M arrest by the increased p21 expression in a p53-dependent manner, suppress the loss of contact inhibition and represses activation of the hypoxia inducible factor, which plays an important role in the progression of tumorigenesis. However, seldom studies about ING4 inducing tumor cells apoptosis were reported.The C6 cells (mouse glioma cells) were infected respectively with the blank adenovirus carrying GFP (Ad) and the recombinated Ad-hING4-His, then RT-PCR assay was used to detect the transcriptions of hING4, as well Western-blotting assay was ued to detect the expressions of hING4. The effects of hING4 expression upon C6 cells were observed, and the growth curve was drawed and tumor control rates were calculated. The C6 cells, which were affected by blank Ad and Ad-hING4-His, were respectively observed by LSCM (laser scan confocal microscope) and transmission electron microscope (TEM), detected by flow cytometry; and the genomic DNA of both groups were extracted and electrophoresised in agarose gel to examinate the DNA fragments. The results showed hING4 can significantly inhibit the growth of C6 cells by promoting the cell’s apoptosis, which probably is the first one to prove this property of ING4.The experimental and theoretical foundation for gene therapy for gliomas with ING4 in the future was established.

Anti-Müllerian hormone (AMH) is a member of the transforming growth factor (TGF)-beta superfamily and mainly expressed by the granulosa cells of ovarian follicles. In women AMH is only expressed in ovarian follicles and therefore can be used for the evaluation of the ovarian reserve function and the prediction of ovary ageing and ovarian response during in vitro fertilization (IVF) treatment. This article summarizes the clinical application of AMH, especially in evaluating ovarian reserve functions.
Anti-Mullerian Hormone ; blood ; Biomarkers ; blood ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; blood ; Humans ; Ovarian Follicle ; physiology ; Ovarian Hyperstimulation Syndrome ; prevention & control ; Ovary ; physiology ; Polycystic Ovary Syndrome ; blood

Objective To evaluate the clinical value of double-balloon enteroscopy( DBE)in as-sessing the effect of mucosal healing in patients with moderate small bowel Crohn's disease( CD)treated with low-dose azathioprine. Methods CD patients who were naive to any immunomodulators or biological a-gents with lesions mainly located in ileu were screened by multislice CT enterography and anal-route DBE at baseline. Lesions at 150 cm proximal to ileocecal valve were assessed by DBE with Simple Endoscopic Score for CD( SES-CD)after 12 and 24 months of low-dose azathioprine treatment,respectively. Results A total of 36 patients were enrolled and the average tolerated dose of azathioprine was(61. 8 ± 17. 2)mg/day. The total rates of complete,near-complete,partial and no mucosal healing in 36 patients were 19. 4%(7/36), 5. 6%(2/36),27. 8%(10/36),and 47. 2%(17/36)at month 12 and 30. 6%(11/36),25. 0%(9/36), 33. 3%(12/36),and 11. 1%(4/36)at month 24,respectively. The baseline SES-CD score(OR=2. 71, 95%CI:1. 11-6. 63,P=0. 029)and duration of disease(OR=1. 27,95%CI:1. 10-1. 47,P =0. 001) were two relevant factors associated with mucosal healing of small bowel CD. Conclusion DBE has a signif-icant advantage in assessing post-therapy mucosal healing for patients with small bowel CD. The optimal time point for the first follow-up by DBE is at least 12 months after low-dose azathioprine treatment.

The biological!activities i.e. antineoplastic activities and antiviral activity of the two novel kinds of interferons: hIFN-?1 and hIFN-? were studied and compared. First the fusion expression vector: pcDNA3.1A-hIFN-?1-His and pcDNA3.1A-hIFN-?-His by PCR was constructed,then the two kinds of plasmids were transfected into WI-38 (human embryonic lung cells) with liposome. And cytopathic effect (CPE) suppression test was used to study and compare the antiviral activities of rhIFN-?1-His and rhIFN-?-His, meanwhile MTT assay was used to detect their antineoplastic activities.It was found that, antiproliferative activity and MxA protein induction shown by rhIFN-?1-His is more powerful than of rhIFN-?-His. The antiviral molecular mechanisms of both hIFN-?1 and hIFN-? are related to MxA.The foundation for further study on the bioactivities and mechanism of action of hIFN-?1 and hIFN-? was established.

OBJECTIVETo observe the effect of recombinant adenovirus Ad-ING4 on K562 cells.

METHODSHuman ING4 recombinant transfer vector pAdTrack-CMV-ING4 was constructed by enzyme digest and ligation of human ING4 gene which was obtained through site specific point mutation of mouse ING4. The vector was co-transduced into BJ5183 E. coli with pAdEasy-1. The new recombinant adenovirus vector pAdEasy-1-pAdTrack-CMV-hING4 was transfected into QBI-293A cells. To obtain the ING4 recombined adenovirus (Ad-ING4). Ad-ING4 was used to infect K562 cells. The effect on K562 cells of ING4 was tested by LSCM FCM and immunohistochemistry.

RESULTSHuman ING4 recombinant adenovirus vector was constructed successfully, and high titre ING4 recombinant adenovirus (Ad-ING4) was obtained. ING4 can down-regulate the expression of bcl-2 and up-regulate expression of bax. The apoptosis of K562 cells induced by ING4 was proved by LSCM FCM and immunohistochemistry. The apoptosis rate was 19.7% (after 72h), which displayed significant difference compared with that of control groups (P < 0.01).

CONCLUSIONAd-ING4 can inhibit the growth of K562 cells and induce the cells apoptosis. The human ING4 recombinant adenoviral vector constructed might provide an approach to the target therapy of tumors.

Adenoviridae ; genetics ; Animals ; Apoptosis ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; Cell Cycle Proteins ; genetics ; Cell Proliferation ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; K562 Cells ; Mice ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Plasmids ; genetics ; Transfection ; Transformation, Bacterial ; Tumor Suppressor Proteins ; genetics

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.
Apigenin ; analysis ; Chromatography ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Flavanones ; analysis ; Flavonoids ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Kaempferols ; analysis ; Luteolin ; analysis ; Magnoliopsida ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Silica Gel

The human interleukin-17F(hIL-17F) gene was amplified by RT-PCR from PHA-activated human peripheral blood mononuclear cells (PBMCs). It was then subcloned into the retrovirus vector pSIV-1. The pSIV-1/hIL-17F together with its two-helper virus vectors pHIT456 and pHIT60 cotransfected into the package cell 293T by lipofectin to produce mature recombinant retrovirus, which was then used to infect SMMC-7721 hepatocarcinoma cells (HCCs), and the cells were selected in the presence of G418. The integration, transcription, expression of hIL-17F gene in SMMC-7721 cells was identified by PCR, RT-PCR and Western blot respectively. MTT and FCM showed that hIL-17F couldn't alter the proliferation and cell cycle of SMMC-7721 cells, but ELISA showed that it could down-regulate IL-6, IL-8 and VEGF expression. The effect of rhIL-17F supernatant on growth suppressing of ECV304 cells was observed by MTT. The experiment of human hepatocarcinoma xenograft tumor in nude mice showed that the formation and growth rates of hIL-17F-transgenic SMMC-7721 showed an obvious decline, and VEGF and CD34 expression and angiogenesis of the transgenic neoplasms was also evidently defined. hIL-17F can markedly inhibit the growth of human hepatocarcinoma xenograft tumor in nude mice by antiangiogenesis. This study provided an experimental evidence for further conducting tumor gene therapy by targeting vascularity and exploiting antiangiogenic novel medicine related to hIL-17F.
Animals ; Cell Line, Tumor ; Cell Proliferation ; Genetic Therapy ; Humans ; Interleukin-17 ; genetics ; Liver Neoplasms, Experimental ; pathology ; therapy ; Mice ; Mice, Nude ; Retroviridae ; genetics ; Vascular Endothelial Growth Factor A ; analysis ; Xenograft Model Antitumor Assays