CCNB1 may as a biomarker for the adipogenic differentiation of adipose-derived stem cells in the postoperative fat transplantation of breast cancer.

Background: Adipose-derived stem cells (ADSCs) are closely associated with the survival rate of transplanted fat in breast reconstruction after breast cancer surgery. Nevertheless, the intrinsic mechanisms regulating ADSCs adipogenic differentiation remain ambiguous. The aim of our study was to explore the relevant genes and pathways to elucidate the potential mechanisms of adipogenic differentiation in ADSCs. Methods: The Gene Expression Omnibus (GEO) dataset GSE61302 was downloaded and analyzed to identify differentially expressed genes (DEGs). Key genes and signaling pathways were obtained through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional and enrichment analysis. Protein-protein interaction (PPI) network and hub gene analyses were performed with the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database and Cytoscape software. Finally, the transcription levels of hub genes in the adipogenic differentiated group and undifferentiated group of ADSCs were compared via real-time quantitative polymerase chain reaction (RT-qPCR). Results: In total, 1,091 DEGs were identified through bioinformatics analysis of the adipogenic differentiated group and undifferentiated group. If was then found that the 10 downregulated key genes, CCNB1, NUSAP1, DLGAP5, TTK, CCNB2, KIF23, BUB1B, CDC20, CDCA8, and KIF11 may play important roles in the adipogenic differentiation of ADSCs. Subsequent in vitro experimental verification also revealed that the messenger RNA (mRNA) expression levels of cyclin B1 in adipogenic differentiated cells and undifferentiated cells were significantly different at the early stage (P<0.05), but there was no significant difference at the late stage (P>0.05). Conclusions: As a key gene, CCNB1 might be a potential biomarker in the adipogenic differentiation of ADSCs at the early stage.

[Effect of transcutaneous auricular vagus nerve stimulation on the expressions of GFAP and MAP2 in ischemic penumbra of rats with middle cerebral artery ischemia].

OBJECTIVE: To observe the effects of transcutaneous auricular vagus nerve stimulation (taVNS) on the motor function and the expression of glial fibrillary acidic protein (GFAP) and microtubule associated protein 2 (MAP2) in cerebral ischemic penumbra of rats with middle cerebral artery occlusion (MCAO) and explore the mechanism of taVNS in the improvement of motor function in MCAO rats. METHODS: A total of 48 male SD rats were randomized into a sham-operation group, a model group, a transcutaneous auricular non-vagus nerve stimulation (tnVNS) group and a taVNS group, with 12 rats in each group. The suture-occluded method was adopted to prepare MCAO rat model. The auricular rim was stimulated in the tnVNS group and the concha stimulated in the taVNS group, 2 mA in intensity, 10 Hz in frequency, 30 min each time, once a day, for 14 days consecutively. The nerve functional assessment was recorded in each group. The expressions of nicotinic acetylcholine receptor (α7nAchR) in the cerebral ischemic penumbra and the spleen were detected by using Western blot. With the immunofluorescence, the expressions of GFAP and MAP2 were detected. RESULTS: After modeling, compared with the sham-operation group, the nerve functional score was increased in the model group, the tnVNS group and the taVNS group (P<0.01), suggesting the success of modeling. After treatment, the score was increased in the model group (P<0.01) as compared with the sham-operation group. Compared with the model group, the neurological deficit score was reduced in the taVNS group (P<0.01). Compared with the sham-operation group, GFAP expression was increased and MAP2 expression was reduced remarkably in the cerebral ischemic penumbra in the model group (P<0.05). In comparison with the model group, GFAP expression was reduced, while MAP2 expression was increased remarkably in the cerebral ischemic penumbra in the taVNS group (P<0.05). There were no significant differences in the abovementioned indexes between the model group and tnVNS group (P>0.05). The differences in the expression of α7nAchR in the cerebral ischemic penumbra and the spleen had no statistical significance among groups (P>0.05). CONCLUSION: TaVNS is effective on neuroprotection in MCAO rats, which may be related to its function of inhibition of GFAP expression and promotion of MAP2 expression in the ischemic penumbra.

The mechanisms through which auricular vagus nerve stimulation protects against cerebral ischemia/reperfusion injury.

Previous studies have shown that vagus nerve stimulation can improve patients' locomotor function. The stimulation of the auricular vagus nerve, which is the only superficial branch of the vagus nerve, may have similar effects to vagus nerve stimulation. However, the precise mechanisms remain poorly understood. In this study, rat models of cerebral ischemia/reperfusion injury were established by modified Longa ligation. Twenty-four hours later, 7-day auricular vagus nerve stimulation was performed. The results showed that auricular vagus nerve stimulation promoted the secretion of acetylcholine, inhibited the secretion of interleukin-1ß, interleukin-6, and tumor necrosis factor-α, and reduced connexin 43 phosphorylation in the ischemic penumbra and motor cortex, promoting locomotor function recovery in rats with cerebral ischemia/reperfusion injury. These findings suggested that auricular vagus nerve stimulation promotes the recovery of locomotor function in rats with cerebral ischemia/reperfusion injury by altering the secretion of acetylcholine and inflammatory factors and the phosphorylation of connexin 43. This study was approved by the Animal Use and Management Committee of Shanghai University of Traditional Chinese Medicine on November 8, 2019 (approval No. PZSHUTCM191108014).

RP11­156L14.1 regulates SSR1 expression by competitively binding to miR­548ao­3p in hypopharyngeal squamous cell carcinoma.

Emerging studies have demonstrated that long non­coding RNAs (lncRNAs) play essential roles in tumorigenesis. However, the role and function of lncRNAs in hypopharyngeal squamous cell carcinoma (HSCC) have not been completely elucidated. The present study explored the function of a novel lncRNA, RP11­156L14.1, in HSCC. RP11­156L14.1 was revealed to be highly expressed in HSCC tissues and cell lines. Knockdown of RP11­156L14.1 inhibited proliferation, migration, and invasion in HSCC cells. Furthermore, RP11­156L14.1 regulated epithelial­mesenchymal transition (EMT) by controlling EMT­related protein expression. Mechanistically, RP11­156L14.1 exerted its function as a competing endogenous RNA (ceRNA) and directly interacted with miR­548ao­3p. The present study also demonstrated that miR­548ao­3p regulated signal sequence receptor subunit 1 (SSR1) expression by targeting SSR1 3'­UTR. Moreover, the xenograft HSCC tumor model revealed that knockdown of RP11­156L14.1 markedly suppressed HSCC tumor growth in vivo. In summary, these findings indicated that the lncRNA RP11­156L14.1 functions as an oncogene in HSCC by competing with miR­548ao­3p in regulating SSR1 expression. The RP11­156L14.1/miR­548ao­3p/SSR1 axis could be utilized as a potential novel biomarker and therapeutic target for HSCC.

On the networking synthesis of studio factors to the integration of design pedagogy.

Studio is critically important for design education, but few attempts have been made to demonstrate the parallels between studio factors and design performance. This paper adopts a coherent set of analyses to investigate the major studio factors and attempts to quantify the networking interactions among them. First, it describes how architectural studio is usually organised based on some major factors. Next, a theoretical model is established according to the described hypotheses and their mutual interactions. Third, the research method and statistical analysis with structural equation modelling (SEM) are presented. Finally, the results of this empirical examination are presented for discussion and suggestions. Our findings reveal that studio tutorials have no significant effect on undergraduate's design performance. In contrast, students' subjective intention plays a more important role in shaping their behaviour, indicating the importance of transferring those exterior forces into internal benefits when the studio instructor attempts to optimise the pedagogy. These findings are also inspiring for all creative disciplines.

Characterization of Sox9-overexpressing human umbilical cord blood-derived mesenchymal stem cells-based engineered cartilage both in vitro and in vivo.

The source of seed cells is a critical factor for tissue engineering. The goal of this study was to evaluate the chondrogenesis of Sox9-overexpressing human umbilical cord mesenchymal stem cells (hUCMSCs) seeded onto bone matrix gelatin (BMG)/fibrin hybrid scaffolds both in vitro and in vivo. hUCMSCs were stably transfected with Sox9-expressing plasmid and grown on the three-dimensional BMG/fibrin hybrid scaffold for 8 weeks. Scanning electron microscopy and histochemistry were performed. The hUCMSC-loaded scaffolds were implanted into the subcutaneous layer of immunocompetent rats and chondrogenesis and host immune responses were monitored for 8 weeks. We found that hUCMSCs spread well and proliferated from 2 weeks after culturing. They produced abundant glycosaminoglycans and collagen II. At 8 weeks after implanting into rats, the hUCMSCs on the scaffolds formed cartilage-like tissue and displayed positive staining for toluidine blue, safranin O, Masson's trichrome, and collagen II. No significant changes in serum levels of lgG, lgA, lgM, C3, and C4 were observed after implantation of the hUCMSC-loaded scaffolds. Xenogeneic implantation of Sox9-overexpressing hUCMSCs embedded in the BMG/fibrin scaffolds promotes the formation of cartilage-like tissue without inducing evident host immune response. Therefore, Sox9-overexpressing hUCMSCs represent a promising cell candidate for cartilage tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1150-1155, 2017.

Effects of different bronchoalveolar lavage methods on tracheobronchial foreign body patients.

OBJECTIVES/HYPOTHESIS: To compare the different effects of bronchoalveolar lavage (BAL) with diverse combinations of lidocaine, epinephrine, and dexamethasone on pediatric patients with an inhaled tracheobronchial foreign body (TFB). STUDY DESIGN: Randomized controlled study. METHODS: Two hundred forty cases of pediatric patients with inhaled TFB were included in this study, and were randomly divided into four groups using three kinds of drugs for BAL, namely 0.9% saline (S) group, 2% lidocaine with diluted epinephrine (LE) group, 2% lidocaine with diluted epinephrine and 0.5% dexamethasone (LED), control group (C) without BAL. The incidences of intraoperative or postoperative complications and recovery periods were compared. Meanwhile, the concentrations of interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α in BAL fluids and plasma were evaluated by enzyme-linked immunosorbent assay. RESULTS: The incidences of bronchospasm, hypoxemia, and postoperative fever were significantly lower in the LED group than other groups (P < .001). Fever after the TFB removal procedure appeared later in the LED group than the other groups. The improvement and healing periods in the LE and LED groups were significantly shorter than those in the C and S groups (P < .001). The concentrations of IL-1ß, IL-6, and TNF-α in BAL fluids were significantly higher in the LE and LED groups than those in the S group (P < .001), but those in the plasma of the C and S groups were lower compared with the LE and LED groups (P < .001). CONCLUSIONS: BAL with lidocaine, epinephrine, and dexamethasone could promote recovery for TFB patients and reduce incidences of complications, possibly by regulating release of proinflammatory cytokines. LEVEL OF EVIDENCE: 1b.

[Establishment of early warning control charts based on the syndromic surveillance data of outpatient diarrhea in Beijing].

OBJECTIVE: To establish the control charts for early warning of diarrhea based on the syndromic surveillance data from enteric clinic in Beijing. METHODS: The outpatient data from enteric clinic of a Grade Three General hospital in Haidian district, Beijing from April 1 to Oct. 31, 2009 and from May 1 to Nov.10, 2010 were collected, according to the moving average method, the baseline calculated, the value of probability α and µα, the early warning value based on the formula "w=Xj+µαSj" calculated and the early warning control charts drew at last. RESULTS: According to the harmfulness, the severity and controllability of diarrheal diseases, the value of probability α was determined as 0.01, then µα (unilateral) as 2, based on the early warning value, the control charts of diarrheal diseases, bacillary dysentery and other infectious diarrhea were established. CONCLUSION: The enteric clinic requires to further collect baseline data to evaluate and continuously adjust the established control charts for the best early warning model in accordance with the enteric clinic.

MiR-139 targets CXCR4 and inhibits the proliferation and metastasis of laryngeal squamous carcinoma cells.

Our previous studies have showed that chemokine receptor 4 (CXCR4) was over-expressed in laryngeal squamous cell carcinoma (LSCC). However, the mechanism underlying aberrant CXCR4 expression remains unclear. To investigate the roles played by miRNAs in CXCR4 over-expression in LSCC, putative miR-139 was predicted through computational algorithms, including TargetScan, PicTar and miRBase, and luciferase reporter assay was explored to confirm that whether CXCR4 was directly regulated by miR-139. Then, quantitative real-time PCR, immunohistochemistry and in situ hybridization methods were employed to detect the expression of miR-139 and CXCR4 in primary LSCC tissues, normal adjacent mucosal tissues and metastatic lesions derived from 40 LSCC patients in the Second Hospital, Xi'An JiaoTong University. Finally, gain- and loss-of-function assays were adopted to explore the effects of miR-139 and CXCR4 on proliferation, invasion and metastasis of the human LSCC cell line Hep-2 in vitro and in vivo. Our results showed that miR-139 dampened CXCR4 expression, and CXCR4 was directly targeted by miR-139. Additionally, the expression of miR-139 was reduced in alignment with the progression of primary to metastatic LSCC. Moreover, an inverse correlation was observed between miR-139 and CXCR4 protein levels in LSCC specimens. Functional analyses demonstrated that ectopic expression of miR-139 inhibited cell proliferation, migration and metastasis of Hep-2 cells in vitro and in vivo. Similar to the observations seen in restoring miR-139 expression, dampening of CXCR4 expression inhibited cell growth, migration and invasion, whereas miR-139 over-expression reversed the pro-metastatic effect of CXCR4. Taken together, we conclude that miR-139 targets CXCR4 and inhibits proliferation and metastasis of LSCC.

Role of pepsin and pepsinogen: linking laryngopharyngeal reflux with otitis media with effusion in children.

OBJECTIVES/HYPOTHESIS: To analyze the relationship between laryngopharyngeal reflux (LPR) represented by pepsin and pepsinogen, and pathogenesis of otitis media with effusion (OME). STUDY DESIGN: Prospective case-control study. METHODS: Children with OME who required adenoidectomy and tympanostomy/tympanostomy tubes placement were enrolled in OME group, whereas children with adenoid hypertrophy (AH) who required adenoidectomy and individuals who required cochlear implantation (CI) were enrolled in AH and CI groups, respectively. Pepsinogen mRNA and protein levels were assessed by real-time fluorescence-based quantitative polymerase chain reaction and immunohistochemistry in adenoid specimens from the OME and AH groups. Pepsin and pepsinogen concentrations were evaluated by enzyme-linked immunosorbent assay in middle ear fluid and plasma from the OME and CI groups. RESULTS: The levels of pepsinogen protein expressed in cytoplasm of epithelial cells and clearance under epithelial cells in adenoid specimens from the OME group were significantly higher than those in the AH group. Furthermore, the concentrations of pepsin and pepsinogen in the OME group were 51.93±11.58 ng/mL and 728±342.6 ng/mL, respectively, which were significantly higher than those in the CI group (P<.001). In addition, the concentrations of pepsin in dry ears were significantly lower than those in serous and mucus ears in the OME group (F=22.77, P<.001).Finally, the concentration of pepsinogen in middle ear effusion was positively correlated with the expression intensity of pepsinogen protein in cytoplasm of epithelial cells (r=0.73, P<.05) in the OME group. CONCLUSIONS: Pepsin and pepsinogen in middle ear effusion are probably caused by LPR and may be involved in the pathogenesis of OME. LEVEL OF EVIDENCE: 3b.

Isolation and antitumor activities of acidic polysaccharide from Gynostemma pentaphyllum Makino.

Two acidic polysaccharides (GP-B1 and GP-C1) were obtained from Gynostemma pentaphyllum. The molecular weights (Mw) of the two fractions were 79 kDa for GP-B1 and 126 kDa for GP-C1. GP-B1 was composed of Gal, Ara, Man, Rha, Xyl, Glc, GalA and GlcA in a molar ration of 3.5:3.2:0.6:0.9:0.3:0.5:0.6:0.4. GP-C1 consisted of Gal, Ara, Man, Rha, Glc, and GlcA in the proportions of 2.1:1.0:0.3:0.5:0.4:0.9. Among them, GP-B1 treatment had a significant inhibitory effect on the growth of melanoma B16 in vivo and in vitro. Meanwhile GP-B1 could increase the relative spleen weight and stimulate the splenocyte proliferation alone or combined with ConA. Moreover, GP-B1 treatment induced an evident increase in the level of serum TNF-α, IFN-γ, and IL-12 and a reduction for IL-10 production. These results indicate that the antitumor effects of GP-B1 are associated with immunostimulation.

Purification of a polysaccharide from Gynostemma pentaphyllum Makino and its therapeutic advantages for psoriasis.

In current study, a water-soluble polysaccharide (GP-I), with a molecular mass of 33 kDa, was purified from Gynostemma pentaphyllum. Gas chromatography (GC) analysis suggested that it was composed of Glc, Gal, Man, Rha and Ara with a ratio of 5.3: 4.2: 3.0: 0.7: 0.8. The GP-I (25, 50, 100, 200 and 400 µg/ml) was found to have significant anti-proliferative effects on HaCat cells in a dose-dependent manner, as measured by MTT assay. On the contrary, Trypan blue exclusion experiment indicated that GP-I had no cytotoxicity to HaCat cells. Moreover, the decrease of mitochondrial membrane potential (MMP) in GP-I treated cells was also observed, indicating apoptosis in HaCat cells. Besides, tumor necrosis factor-α (TNF-α), a vital pro-inflammatory cytokine in psoriasis, in the supernatant of HaCat cells was dramatically reduced by GP-I. Collectively, these findings suggested that GP-I was a promising agent to be developed for psoriasis treatment in clinical therapy.

[Changes of pathogens for nosocomial infection of patients with hematological diseases].

In order to investigate the distribution of nosocomial infection in patients with hematological diseases in our hospital, and to explore the changes of the pathogens isolated. The method of retrospective investigation and analysis was employed. 1164 strain pathogens were isolated from the patients with hematological diseases during the period of 1997-2009. The results showed that the Gram-positive cocci infection increased gradually during the 13 years, but has been stable in the last 4 years. The Gram-negative bacteria showed a trend decrease. The fungi increased during these years. The rates of infection with gram-positive cocci, gram-negative bacteria and fungus were 28.2%, 59.8% and 12.0% respectively. For the details, Escherichia coli infection rate was the highest: 12.1%, followed by Pseudomonas aeruginosa (9.1%), Enterobacter (8.4%), Klebsiella pneumoniae (7.4%), Staphylococcus epidermidis (6.3%) and Enterococci (6.6%). The distribution of G(+)- and G⁻ pathogens showed obvious change on end of 1990's and beginning of this century, but it was tending towards stability on recent years; the incidence of fungus was tending towards increase, which was related to wide application of strong broad-spectrum antibiotics. In conclusion, the patients with hematological diseases, as the high-risk group of nosocomial infection, should be monitored strictly. Infection is related to many factors, and the main factor is dysfunction of autoimmunity. The strategies should be explored to strengthen the immune protection and set up a reasonable scheme of antibiotics.

Lentivirus-mediated knockdown of aggrecanase-1 and -2 promotes chondrocyte-engineered cartilage formation in vitro.

Chondrocyte-based tissue engineering has emerged as a promising approach for repair of injured cartilage tissues that have a poor self-healing capacity. However, this technique faces a major limitation: dedifferentiation of chondrocytes occurs following several passages in culture. Aggrecan, a major component of cartilage extracellular matrix, plays an essential role in chondrocyte differentiation. The aim of this study is to determine whether inhibition of chondrocyte aggrecanases, key degradative enzymes for aggrecan in cartilage, could benefit chondrocyte differentiation and the preservation of chondrocyte phenotype within a long-term period. Lentivirus-mediated RNA interference (RNAi) was employed to target both aggrecanase-1 and -2 in primary rat chondrocytes, and the transduced cells were seeded into chitosan-gelatin three-dimensional scaffolds. Histological, morphological, and biochemical analyses were performed at 1-8 weeks post-implantation to study chondrocyte survival, differentiation, and function. We found that lentivirus-mediated RNAi notably decreased the abundance of aggrecanase transcripts in chondrocytes but did not affect cell viability. Most importantly, compared to the control constructs seeded with untransduced chondrocytes, the aggrecanase inhibition increased chondrocyte proliferation and reinforced the production of glycosaminoglycans and total collagen, indicative of chondrocyte differentiation. The mRNA expression of chondrocyte marker genes (collagen II and aggrecan) was enhanced by aggrecanase silencing relative to the control. Together our data demonstrate that inhibition of endogenous aggrecanases facilitates chondrocyte differentiation and chondrocyte-engineered cartilage formation in vitro. The combination of lentiviral delivery system and genetic manipulation techniques provides a useful tool for modulation of chondrocyte phenotype in cartilage engineering.

[Analysis of disease types of outpatients with intestinal diseases from two grade-three general hospitals in Beijing].

OBJECTIVE: To understand the spectrum of diseases and epidemiological characteristics of outpatients at Entric Disease Clinic, with a focus on analysis of the distribution of infectious diarrheal diseases in different populations and to explore disease control strategies on Enteric Infectious Diseases for focal groups. METHODS: A census on outpatients at Entric Disease Clinics was conducted in two class Three comprehensive hospitals in Beijing from April 1 to October 31, 2009 based on a descriptive study using diarrhea-syndrome surveillance system set in the two clinics, thus to depict the spectrum of diseases and epidemiological characteristics of outpatients, and analyze the proportion of infectious diarrhea in diarrheal diseases specifically and the rate changes of infectious diarrhea in different months, age groups and occupational groups. RESULTS: Diseases are varied at the two enteric diseases clinics among the patients and there are mainly 10 kinds of diseases, "non-infectious diarrhea" accounted for the highest percentage (77.4%), followed by "unspecified diarrhea" (11.7%), and infectious diarrhea accounted for the least proportion(8.7%)."Gastroenteritis and enteritis" are the most frequently diagnosed cases among all the diseases, was a total of 7 565 cases, accounting for 70.2%. The volume of visits reached its top during summer and autumn(July to September), and the mean volume of visits in this period is (60.78+/-16.85) cases/day. The volume of visits has an obvious seasonal trend, and visits during July and August are the most frequent (41.82% altogether). Patients with "infectious diarrhea" had a highest ratio(5.3%) in May and lowest (1.1%) in October while patients with "bacillary dysentery" accounted for a highest ratio(8.2%) in September and lowest(3.8%) in April. Outpatients are mainly from Beijing city(61.9%), in which young and middle-aged people accounted for 73.9% in total, and student is the main occupation (28.8%). The distributions of diarrheal diseases are the same in different age groups but differ from different occupational groups. Infectious diarrhea accounted for a highest proportion(9.2%) in 18-to-44-year-old age group when using age grouping, and a highest proportion(15.2%) in restaurant service personnel when using occupation grouping. CONCLUSION: The volume of outpatients attended at general hospitals is overwhelming especially in July and August, and the major type is "non-infectious diseases", which indicates an arduous task on prevention and control of Enteric Infectious Diseases. Infectious diarrhea took up a certain amount, but the rate is not that high, which indicates possible missing diagnosis of patients with infectious diseases.Our focal groups would be young and middle-aged people and students in the city. Therefore, the need to extend the consultation hours is urgent. Meanwhile, for the main goal of surveillance and early warning on Enteric Infectious Diseases, all aspects of construction of Enteric Disease Clinic should be strengthened, such as enhance laboratory tests on pathogens, improve diagnosis level of physicians and etc.

Cartilage tissue engineering with demineralized bone matrix gelatin and fibrin glue hybrid scaffold: an in vitro study.

To develop a cartilage-like tissue with hybrid scaffolds of demineralized bone matrix gelatin (BMG) and fibrin, rabbit chondrocytes were cultured on hybrid fibrin/BMG scaffolds in vitro. BMG scaffolds were carefully soaked in a chondrocyte-fibrin suspension, which was polymerized by submerging the constructs into thrombin-calcium chloride solution. Engineered cartilage-like tissue grown on the scaffolds was characterized by histology, immunolocalization, scanning electron microscopy, biochemical assays, and analysis of gene expression at different time points of the in vitro culture. The presence of proteoglycan in the fibrin/BMG hybrid constructs was confirmed by positive toluidine blue and alcian blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene-encoded cartilage-specific markers, collagen type II, and aggrecan core protein. The glycosaminoglycan production and hydroxyproline content of tissue grown on the fibrin/BMG hybrid scaffolds were higher than that of the BMG group. In conclusion, the fibrin/BMG hybrid scaffolds may serve as a potential cell delivery vehicle and a structural basis for cartilage tissue engineering.

Fluorine-induced apoptosis and lipid peroxidation in human hair follicles in vitro.

Fluoride is an essential trace element for human body; however, exposure to high amounts of fluoride has been documented to be correlated with an increasing risk of hair loss. To date, little is known about the mechanism(s) of how fluoride affects hair follicles. Here, we demonstrated that middle (1.0 mmol/L) and high (10.0 mmol/L) concentrations of sodium fluoride (NaF) significantly inhibited hair follicle elongation in vitro, but low NaF (0.1 mmol/L) showed little influence. Moreover, treatment with high levels of NaF resulted in a marked increase in terminal dUTP nick end labeling-positive cells in the outer layer of the outer root sheath, the dermal sheath, and the lower bulb matrix surrounding dermal papilla. Furthermore, the enhanced apoptosis was coupled with an increased oxidative stress manifested as higher malondialdehyde content. Additionally, the presence of selenium considerably antagonized the effects of middle NaF on hair follicles, with regard to either the suppression of hair growth or the induction of oxidative stress and apoptosis. In conclusion, exposure to high levels of fluoride compromises hair follicle growth and accelerate cell apoptosis in vitro. The toxicity of fluoride can be reduced by selenium, at least partially via the suppression of intracellular oxidative stress.

[Effects of RNA interference against aggrecanase 1 gene on extracellular matrix metabolism of cultured chondrocytes in vitro].

OBJECTIVE: To study the effect of RNA interference (RNAi)-mediated aggrecanase-1 gene silencing on extracellular matrix metabolism of cultured rat costochondral chondrocytes. METHODS: Rat costochondral chondrocyte monolayers were obtained by microdissection and digestion. The growth and morphological changes of the chondrocytes were observed after RNAi of aggrecanase-1 gene. The mRNA expression of aggrecanase-1 was detected by RT-PCR method, and aggrecan content was determined by Western blotting. RESULTS: The specific inhibition of aggrecanase-1 by RNAi produced no adverse effect on the morphology and growth of the chondrocytes. The mRNA of aggrecanase-1 decreased and aggrecan content increased significantly after transfection of the chondrocytes. CONCLUSION: Inhibition of aggrecanase-1 decreases aggrecan degradation in cultured rat chondrocytes. RNAi technique can be a useful means for studying extracellular matrix metabolism in the cartilage.

Effects of RNAi-mediated inhibition of aggrecanase-1 and aggrecanase-2 on rat costochondral chondrocytes in vitro.

AIM: Failure of transplanted cartilage or allogenic chondrocytes is attributed mainly to immunological rejection and cartilage degradation. A major feature is the loss of aggrecan from the cartilage matrix, primarily due to the action of the specific proteinases aggrecanase-1 and aggrecanase-2. The aim of this in vitro study was to determine whether the specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi would mitigate aggrecan loss from cultured chondrocytes. METHODS: Expression plasmid vectors of shRNA targeting aggrecanase-1 and aggrecanase-2 were constructed and transfected into cultured rattus costochondral chondrocytes. The transfected cells were induced with interleukin-1beta (IL-1beta). Gene mRNA levels were analyzed by RT-PCR. Aggrecan and collagen II content were measured by immunohistochemistry and Western blotting. RESULTS: As the chondrocytes underwent dedifferentiation, aggrecanase-1 increased significantly. The specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi had no negative effect on the morphology and growth velocity of the chondrocytes. The mRNA of aggrecanase-1 and aggrecanase-2 decreased significantly. The alpha-2-macroglobulin expression level was increased by the shRNA specific for aggrecanase-1. Other genes of the chondrocytic extracellular matrix were not affected. RNAi significantly increased the aggrecan and collagen II content of chondrocytes treated with IL-1beta. CONCLUSION: The results suggest that inhibition of aggrecanase-1 and aggrecanase-2 by RNAi can mitigate aggrecan degradation, without interfering with chondrocytic gene phenotype recovery. RNAi technology can be a useful tool for studying degenerative processes in cartilage.

[Investigation and analysis of neonate deformity in water arsenic exposure areas].

OBJECTIVE: To explore the level and feature of neonate deformity in water arsenic exposure areas, as to finding out an evidence for the study and prevention of the arsenic exposure. METHODS: The birth situation of neonate was surveyed from 1998 to 2004 in water arsenic exposure areas according to cross-sectional survey. The results were classified in accordance with ICD-10 and common surveillance of china. The population of Shanyin County served as the common people and the data were analyzed by SPSS 11.5 for windows. RESULTS: The neonates surveyed were 2467 cases. There were 49 neonates deformity found in this investigation, giving a neonate deformity rate of 198.62 per 10,000 cases, which was shown significantly higher in water arsenic exposure areas than in the normal (U = 3.23, P < 0.01), with types of nervous system deformity, limbs deformity and congenital heart disease as in system classification. There was no significant difference of deformity rate in different sex neonates (chi2 = 0.32, P > 0.05). CONCLUSION: The drinking high-arsenic water over a long period of time should be a risk factor of neonate deformity. Prevention and treatment of endemic arsenic exposure should be urgently needed.