OBJECTIVETo study the effect on anti-respiratory syncytial virus of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro.

METHODSActive component of herba patriniae (AP3) was extracted and its anti-respiratory syncytial virus (RSV) effect was tested. A water soluble substance (AP3) was isolated from a Chinese herb Herba patriniae, by hot water extraction, ethol precipitation and gel permeation column chromatography. The cytotoxicity of AP3 was tested by adding it to HeLa cells directly. Its effect against RSV was estimated by CPEI assay while ribavirin was used as positive control.

RESULTSChemical test showed that the nature of substance AP3 was polysaccharide. The median cytotoxic concentration (TC(50)) of AP3 was 11.45 mg/ml by morphological observation and the median effective concentration (50% effective concentration, EC(50)) of it against replication of the long strain of RSV in HeLa cells was 0.0986 mg/ml. The Therapeutic index (TI = TC(50)/EC(50)) of AP3 was 116.12, much higher than the TI of herba patriniae (AP1) (TI = 59.26) and ribavirin (TI = 53.45). Moreover, AP3 gave a dose-dependent response in inhibiting RSV. In the assay, the effect of AP3 against RSV growth was also tested. In addition, the effect of AP3 on virus growth, AP3 inhibited replication of RSV in HeLa cells, when added at 0 h, 2 h, 4 h after virus infection, were also tested.

CONCLUSIONThis study suggested that the AP3 exerted an obvious inhibitory effect to RSV in HeLa cell culture. This study furnished a reliable evidence for development of a new antiviral drug.

Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Respiratory Syncytial Viruses ; drug effects ; Time Factors

OBJECTIVETo evaluate the effect of human hair keratin (HHK) in peripheral nerve repair and explore the mechanism of sciatic nerve regeneration.

METHODSRat models of sciatic nerve damage was established by creating a 10-mm gap in the sciatic nerve, which was bridged with a HHK implant. Histological examinations of the nerve tissues were performed at different time points after the surgery.

RESULTSDuring the period from 2 days to 2 weeks following HHK implantation, Schwann cells were found to undergo dedifferentiation and proliferate along the HHK implant. Three weeks after HHK implantation, numerous macrophages and megakaryocytes occurred around the HHK, and a large quantity of regenerated Schwann cells aligned in orderly fashion was seen between the fine filaments of partially degraded HHK, where axons and capillaries were also observed. Six weeks later, massive nerve fibers and capillaries developed around the HHK, and at 9 weeks, the HHK implant was substantially degraded and numerous regenerated nerve fibers occurred characterized by obvious epineurium and perineurium. Till 12 weeks after HHK implantation, HHK was almost completely degraded and replaced by the newly regenerated nerve fibers that had grown across the nerve defect.

CONCLUSIONSHHK is an ideal material for nerve injury repair. Apocytosis plays a key role in the differentiation process of highly differentiated Schwann cells into immature Schwann cells following nerve injury. As a protective mechanism, the axons undergo enclosure and dissociation following injuries, and the intact axons give rise to growth cones that extend fibers of growing buds to competitively bind the one or more Schwann cells, but only one such but finally develops into a complete axon. The nerve fiber barrier membrane is derived from the capillary menchymal stem cells and the outmost vascular barrier membrane. The regeneration of the Schwann cells, axons and the nerve membrane is the result of self-organization through a well synchronized and coordinated mechanism.

Animals ; Female ; Hair ; chemistry ; Humans ; Keratins ; administration & dosage ; pharmacology ; Male ; Nerve Regeneration ; drug effects ; physiology ; Prostheses and Implants ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; physiopathology

We evaluated the ability of lactic acid bacteria, Weissella cibaria, isolated from the oral cavity to adhere to epithelial cells. W. cibaria efficiently adhered to KB cells and HeLa cells. In addition, W. cibaria efficiently adhered to Fusobacterium nucleatum. But the adhesiveness of W. cibaria disappeared upon exposure to LiCl or pronase, suggesting that the S-layer proteins of W. cibaria mediated the adhesiveness. The molecular mass of the S-layer proteins extracted from W. cibaria was approximately 50 kDa. When W. cibaria strains were washed with 0.45% saline, the bacteria were efficiently adhered to the epithelial cells. In conclusion, W. cibaria has the ability to adhere to epithelial cells through the S-layer proteins.
Adhesiveness ; Bacteria ; Epithelial Cells* ; Fusobacterium nucleatum ; HeLa Cells ; Humans ; KB Cells ; Lactic Acid ; Mouth ; Pronase ; Weissella*

BACKGROUND: Although the pathogenesis of vitiligo isn't fully understood, a recent study demonstrates that oxidative stress plays an important role to induce vitiligo. Peroxiredoxin (Prx) is a novel peroxidase family to remove hydrogen peroxide using thioredoxin system, which is consisted of thioredoxin, thioredoxin reductase, and NADPH. OBJECTIVE: This study aimed to investigate the change of expression of Prx I to elucidate the role of oxidative stress in the pathogenesis of vitiligo. METHODS: Sample specimens were obtained from the lesional skin of vitiligo patients, and non-depigmented skin was obtained from the perilesional area as control samples. The skin samples were immediately frozen using liquid nitrogen, and then section samples were prepared to perform immunohistochemical staining with antibodies for Prx I. Some of the skin biopsy samples were used for primary culture of keratinocytes. Protein extracts from the expanded keratinocytes were prepared for Western blot analysis of Prx I. RESULTS: In vitiligo, the ubiquitous expression of Prx I in all layers of epidermis, which was also observed in the normal perilesional skin, was reduced in the depigmented lesion of vitiligo patients. The reduction of Prx I was remarkable from the lesions which were exposed to sunlight. Consistently, Prx I expression from the lesional keratinocytes were noticeably reduced in comparison with that from perilesional keratinocytes. CONCLUSION: Our results showing that Prx I is impaired in the epidermis of depigmented lesions of vitiligo patients suggest that oxidative stress is an important factor to induce vitiligo.
Antibodies ; Biopsy ; Blotting, Western ; Epidermis ; Humans ; Hydrogen Peroxide ; Keratinocytes ; Nitrogen ; Oxidative Stress ; Peroxidase ; Peroxiredoxins ; Skin ; Sunlight ; Thioredoxin-Disulfide Reductase ; Thioredoxins ; Vitiligo

BACKGROUND: Tissue inflammation and remodeling have been extensively studied in various tumors in relation with their invasiveness and metastasis. OBJECTIVE: The purpose of this study was to investigate the change in tissue inflammation and remodeling markers in cutaneous squamous cell carcinoma (SCC). METHODS: Expression levels of cyclooxygenase-2 (COX-2) as an inflammatory marker and matrix metalloproteinases-2 and -9 (MMPs 2/9) as remodeling markers were studied in mouse and human SCCs. Western blot analysis and RT-PCR for COX-2 and MMPs 2/9 were performed with skin samples from SCC patients and chronic ultraviolet B (UVB)-induced SCC from hairless mice. RESULTS: mRNA and protein levels of COX-2 and MMPs 2/9 were up-regulated with the higher sensitivity for MMP-9 in mouse SCCs, which were induced by chronic UVB irradiation. Consistently, COX-2 and MMPs 2/9 were up-regulated with the higher sensitivity for MMP-9 in human SCCs. CONCLUSION: COX-2 and MMPs 2/9 are up-regulated in well-differentiated cutanous SCC. Our findings indicate that inflammatory and tissue remodeling processes are actively induced during carcinogenesis of cutaneous SCC.
Animals ; Blotting, Western ; Carcinoma, Squamous Cell ; Cyclooxygenase 2 ; Humans ; Inflammation ; Matrix Metalloproteinases ; Mice ; Prostaglandin-Endoperoxide Synthases ; RNA, Messenger ; Skin ; Up-Regulation

BACKGROUND: Vitiligo is a depigmented disorder, causing serious cosmetic problems for patients. In diagnostic and therapeutic aspects, vitiligo should be differentiated from other hypopigmented disorders as the therapeutic approach and prognosis are different for each disease. OBJECTIVE: This study aimed to compare the usefulness of several markers for melanocytes or melanin in differential diagnosis of vitiligo. METHODS: Twenty-eight patients were studied, who were diagnosed clinically as suffering from one of the following diseases: vitiligo, nevus depigmentosus, pityriasis alba, postinflammatory hypopigmentation, and idiopathic guttate hypomelanosis. Skin samples (frozen or paraffin-fixed) were obtained from depigmented patches and normal neighboring skin (control). Histological staining was performed by using Fontana-Masson, S-100, MART-1, and DOPA. The staining level of lesional skin was compared with that of normal skin. RESULTS: When the staining level of vitiligo was compared with that of others, vitiligo was significantly lower in Fontana-Masson (13.3+/-17.2% vs 44.4+/-23.7%), S-100 (49.5+/-14.9% vs 74.7+/-24.2%), MART-1 (7.4+/-8.7% vs 68+/-33.9%), and DOPA (9.5+/-11.3% vs 58.2+/-29.5%) (p<0.0001). CONCLUSION: MART-1 and DOPA are valuable markers in differential diagnosis of vitiligo. However, Fontana-Masson, a marker of melanin, had some limits in detecting melanocytes, and S-100 showed non-specific staining other than melanocytes.
Cosmetics ; Diagnosis, Differential ; Dihydroxyphenylalanine ; Humans ; Hypopigmentation ; Melanins ; Melanocytes ; Nevus ; Pityriasis ; Prognosis ; Skin ; Stress, Psychological ; Vitiligo

The present study was designed to isolate and characterize novel chemical constituents of the stem bark of Juglans mandshurica Maxim. (Juglandaceae). The chemical constituents were isolated and purified by various chromatographic techniques. The structures of the compounds were elucidated on the basis of spectral data (1D and 2D NMR, HR-ESI-MS, CD, UV, and IR) and by the comparisons of spectroscopic data with the reported values in the literatures. Two long chain polyunsaturated fatty acids (1 and 2) were obtained and identified as (S)-(8E,10E)-12-hydroxy-7-oxo-8,10-octadecadienoic acid (1) and (S)-(8E, 10E)-12-hydroxy-7-oxo-8,10-octadecadienoic acid methyl ester (2). To the best of our knowledge, this is the first report on the isolation and structural elucidation of the two new conjugated ketonic fatty acids from this genus.
Fatty Acids, Unsaturated ; chemistry ; isolation & purification ; Juglans ; chemistry ; Plant Bark ; chemistry ; Spectrum Analysis