Objective To investigate the feasibility of transient transfection of human VEGF165 gene into HaCaT cells and the effects of supernatant from transfected cell culture on pig hair follicles in vitro. Methods PIRES2-EGFP-VEGF165 was transiently transfected into HaCaT cells with lipofectamine?2000. Expression of EGFP( enhanced green fluorescence protein, EGFP) was observed by laser confocal microscopy. VEGF in the supernatant was detected by ELISA. Further,the supernatant was added to pig hair follicles cultured in vitro. The growth and morphologic changes of hair follicles were measured. Results PIRES2-EGFP?VEGF165 was successfully transfected into HaCaT cells, which confirmed by laser confocal microscopy and ELISA. Moreover, the supernatant of HaCaT cells transfected with PIRES2-EGFP-VEGF165 could accelerate the growth of pig hair follicles and prolong their anagen phase. Conclusion PIRES2-EGFP-VEGF165 could be successfully transiently transfected into HaCaT cells, and expressed VEGF could upregulate the growth of pig hair follicles in vitro.

Objective To investigate the effect of water soluble extracts of some traditional Chinese herbs on hair growth of pig hair follicle in vitro. Methods Pig hair follicles were cultured with Williams E medium (control group) or Williams E medium with water soluble extracts of Chinese herbs (experimental group). Hair growth and morphological changes in the hair follicle bulb were observed by microscope, and apoptosis of the pig hair follicle cells was detected by a TUNEL technique. Results On day 7 of culture, the hair growth in the control group was slower than that in the experimental group (P

Objective To observe the morphological changes of hairs in two patients with loose anagen hair syndrome.Methods Light microscopy,scanning electron microscopy and transmission electron microscopy were used to observe the morphology of hair shaft and follicles from two patients,including a 3-year-old girl child and her mother,with loose anagen hair syndrome.Results Light microscopy revealed that hair bulb was deformed,hair shaft was distorted,damaged,and even disrupted,and hairs tapered in diameter at their distal end.Scanning electron microscopy showed deformed or distorted hair shaft and wave-like edge of hair cuticles.Transmission electron microscopy revealed that pathological changes were mainly localized in the inner root sheath with vacuolization in both inner and outer root sheath cells.Intercellular adhesion was weak with a decrease or disappearance of desmosomes.Conclusion The pathological changes of hairs are mainly localized in the inner root sheath in patients with loose anagen hair syndrome.

Objective To investigate the expression of angiogenin in human hair follicle and evaluate its effect on hair growth. Methods Intact anagen hair follicles were isolated from human occipital scalp ob- tained from brain surgery. Some isolated human hair follicles were directly subjected to RT-PCR and im- munohistochcmical method for the detection of the mRNA expression and protein distribution of angiogenin in, respectively; some were cultured and incubated with angiogenin (0-200 ng/mL), and the measurement of hair follicle length was performed before and after 6-day culture. Human dermal papilla cells were isolated from the remaining hair follicles, cultured, and treated with angiogenin ranging from 0 to 200 ng/mL for 48 hours, then, MTT assay was used to detect the cell proliferation, and flow cytometry to analyze the cell cy- cle. Results RT-PCR showed the mRNA expression of angiogenin in human hair follicles, and angiogenin protein was observed with immunohistochemistry at the hair papilla and dermal sheath. The angiogenin (25-200 ng/mL) stimulated the growth of human hair follicles in a dose-dependent manner in vitro (P < 0.05 ). Also, as flow eytometry revealed, the treatment with 12.5-200 ng/mL of angiogenin significantly pro- moted the proliferation of human dermal papilla cells (P<0.05), and increased the percentage of cells in S phase as well as cell proliferation index (both P<0.05). Conclusion Angiogenin may be a novel stimulus for hair growth.

Objective To investigate the feasibility of RNA interference(RNAi)strategy in mouse(embryonic stem)ES cells and study the possibility of RNAi in dermatological research.Methods Specific RNAi was introduced with EGFP gene as a target,either by in situ production of dsRNA from transient trans-fection of a plasmid harboring a547bp inverted repeat,or by direct transfection of dsRNA made by in vitro transcription.Gene silence of EGFP in EGFP-transgenic mouse was investigated by topical delivery of dsRNA.Results This longer dsRNA was capable of inducing a sequence-specific RNAi for the target gene in epi-some and in chromosome of ES cells.It was also shown that this sequence-specific RNAi was observed in EGFP-transgenic mouse by topical delivery of dsRNA.Conclusion These results suggest that ES cell possess RNAi activity by longer dsRNA.We first report the study on RNAi activity by topical delivery of specific dsRNA.Above findings offer a possibility to use dsRNAi for inhibition of gene expression in dermatological re-search.

Objective To evaluate the methods and effect of balloon catheter dilation of esophageal caustic ingestion stenosis in children. Methods We analysed 18 cases, including 10 cases of esophageal stenosis due to ingestion of sulphuric acid, 7 cases of esophageal stenosis caused by ingestion of sodium carbonate and the last one through ingestion of chemical materials include zinc sulphate. Barium esophagogram was taken before dilation for every patient and the balloon size varied from 4 mm?40 mm to 16 mm?40 mm or 20 mm?40 mm in diameter was selected for the procedure. Results 18 cases were all successful in dilation by balloon catheter, without esophageal perforation and other complications. The satisfactory results maintained from six months to thirty months with remarkable improvement clinicoly. Conclusions Balloon catheter dilation is a simple, safe and reliable method for the treatment of esophageal caustic ingestion stenosis in children, and should be recommended as the first choice.

Objective To explore the relationship between serum HBeAg、the qualification of HBV DNA and the liver pathologic change in chronic hepatitis B virus (HBV)carriers with normal liver function.Methods Two hundred and forty-four chronic hepatitis B virus carriers with normal liver function were performed pathology examination by Liver biopsy.Meanwhile liver function,HBV DNA level and serological serum markers of B-hepatitis examination were detected.Results Pathology results showed that,of all 244 cases,7 cases was cirrhosis(2.9％),143 for slight CHB (58.6％),32 for moderate CHB (13.1％) and 9 for severe CHB (3.6 ％).And 53 (21.7％)chronic hepatitis B virus carriers were the normal histology morphology.Fony-eight cases (19.7％) were in inflammation stage G≥2 and 54(22.1％) were with fibrosis stage S≥2.For cases with HBV DNA positive,The inflammation and ftbrosis stages in HBeAg negative group were more severe than that in HBeAg positive group (P ＜ 0.05).The fibrosis stages in patients with low HBV DNA level were severe than that in high HBV DNA levels (P ＜ 0.05).No significant differences were observed in the inflammation stage between subjects with high or low HBV DNA level.Conclusion Most chronic HBV carriers with normal liver function were with different degrees of liver inflammation and fibrosis.HBeAg and HBV DNA associated with liver pathological change.

Objective To screen differentially expressed microRNAs at different stages of hair cycle in a murine model.Methods This study included 30 inbred female C57BL/6 mice (age,6-8 weeks; body weight,15-18 grams).Hair growth cycle was induced in the back skin of C57BL/6 mice by application of wax/rosin followed by depilation under anesthesia witl 1％ chloral hydrate.Three mice were sacrificed by cervical dislocation on day 0,8 and 20 after the induction,and skin tissue was achieved from the same depilated areas parallel to the spine.Total RNAs were extracted from the murine skin and subjected to microarray analysis of microRNA expression.Results Compared with telogen skin,the murine anagen skin showed a higher expression of miR-690,obselote-49 and miR-1308,but a lower expression of miR-291a-5p and miR-212.The expressions of miR-690,obselote-49 and miR-31 were significantly up-regulated,while those of miR-127-3p and miR-212 were downregulated in the catagen skin in comparison with the telogen skin.Conclusion Seven microRNAs are identified in this study to be differentially expressed in murine skin between different stages of hair cycle,which may provide a direction for future research.

Background and purpose:We investigated whether lfuorine-18 lfuorodeoxyglucose (18F-FDG) maximal standard uptake value (SUVmax) of the primary tumor (SUV-T), SUVmax of the regional lymph nodes (SUV-N) or the overall loco-regional lesion SUVmax (SUV-TOTAL) was related to survival of patients with stage Ⅲ non-small cell lung cancer (NSCLC) who received Cetuximab and combined definitive chemoradiotherpay. Methods:From September 2009 to July 2012, seventeen patients with unresectable stageⅢNSCLC receiving cetuximab with cisplatin/vinorelbine (NP) followed by concomitant NP and intensity-modulated radiotherapy (IMRT) at the Fudan University Shanghai Cancer Center were enrolled onto a prospectively study. All patients received positron emission tomography/computerized tomography (PET/CT) scans within 2 weeks before enrolment. Univariate analysis were used to assess the correlation between SUV-T, SUV-N, SUV-TOTAL, gender, age, histology, tumour-node-metastasis (TNM) stage, performance status (PS) as well as smoking status and survival. The factors which showed statistical signiifcance entered into multivariate Cox-regression model. Survival functions of different populations were estimated by Kaplan-Meier method and compared by Log-rank test. Results:In the univariate analysis, SUV-T, SUV-N, SUV-TOTAL, PS and smoking status were prognostic factors. The best cut-off values for SUV-T, SUV-N and SUV-TOTAL were 11, 11 and 20, respectively. Multivariate analysis revealed that SUV-TOTAL (P=0.012), SUV-T (P=0.025), and SUV-N (P=0.033) were independent predictors of survival with hazard ratio (HR) of 14.7, 11.2, and 6.2, respectively. Conclusion:Local, regional and locoregional maximal SUVs deifned by 18F-FDG PET-CT scanning may have a strong correlation with survival in this patients setting, which merits further study.

Objective To screen genes differentially expressed between dermal papilla cells from occipital and vertex scalp of patients with androgenic alopecia (AGA) through a 3D culture system.Methods Dermal papilla cells isolated from the occipital scalp tissue of patients with AGA were cultured in a 2D system for several days.Then,the third-passage dermal papilla cells were subjected to a 3D culture with the presence of dihydrotestosterone (DHT) for 72 hours (experimental group).The dermal papilla cells isolated from the vertex scalp tissue of patients with AGA,which were cultured in a 3D system with dimethyl sulfoxide,but not DHT,served as the control group.Subsequently,total RNA was extracted from the cells and reversely transcribed into cDNA followed by labeling with Cy3 and hybridization to a NimbleGen microarray.Gene ontology (GO) and pathway analysis was carried out to screen differentially expressed genes between the experimental and control group.Real time PCR was conducted to validate the results of microarray analysis.Results As the genome-wide expression profile analysis showed,there were 622 genes differentially expressed between the experimental group and control group,of which,359 were up-regulated and 263 were down-regulated in the experimental group compared with the control group.The above results were corffirmed by real time PCR.GO analysis revealed that the up-regulated genes,such as the CHEK1 and Tobl genes,were mainly involved in the inhibition of cell proliferation and promotion of cell apoptosis,while the down-regulated genes,such as the BAMBI,EFNA3,Dlx3 and UCGC genes,were associated with the acceleration of cell proliferation as well as the growth and development of epidermis.Pathway analysis showed that cell circle-controlling molecules were the most abundant molecules.Conclusions Numerous signalling molecules and pathways are involved in the development of AGA,which are mainly responsible for the modulation of cell circle,proliferation and apoptosis.