Objective To construct yeast cDNA expression library of human dermal papillae cells (DPCs) in primary culture.Methods Human dermal papilla cells (DPCs) were isolated by two-step digestion method and cultured in DMEM medium.Total RNA was extracted from primary DPCs that exhibited an aggregative behavior in culture,then,cDNA was synthesized and amplified by using CloneMinerTM cDNA Library Construction kit to construct primary cDNA library and yeast cDNA expression libary.Results The average titer and total clones were 7.0×106 colony forming units(cfu)/ml and 1.4×107 cfu respectively in the primary library,5.5×106 cfu/ml and 1.1×107 cfu respectively in the yeast expression library.The average insert size Was 1.2 kb and the recombination rate was above 95%.Conclusions The yeast cDNA expression library of DPCs in primary culture has been successfully constructed.which will lay a foundation for screening proteins interacting with HSPC016 gene in DPCs with yeast two-hybrid system.

Objective To implore the characteristics of epithelial-mesenchymal transition(EMT) in human prostate cancer stem progenitor (S/P) cells isolated from LNCaP cell lines.Methods The S/P cells were obtained through florescence-activated cell sorting (FACS).Western blot and immunofluorescence assay were used to detect the S/P cells' EMT markers expression,such as E Cadherin,N Cadherin,Vimentin and Snail.Soft agar assay was used to detect the tumorigenesis ability of S/P cells.Cell migration assay was used to detect the migration ability of S/P cells.Results Compared with non S/P cells,the expressions of EMT markers,such as N Cadherin,Vimentin and Snail,were increased in S/P cells,while the expressions of epithelial marker and E Cadherin were decreased in S/P cells.After cultured for three weeks,S/P cells and non S/P cells both clonally grew.The colony numbers were (18.34±1.21) and (82.27±7.54),respectively (t =8.617,P =0.001).After cultured for 48 hours,the migration cells number was (25.33±5.13) in non S/P cells and (74.33±7.64) in S/P cells (t =7.953,P =O.001).Conclusions Human prostate cancer S/P cells isolated from LNCaP cell line have some characteristics of EMT,such as stronger tumorigenesis and migration ability,which could promote tumor invasion and metastasis.

Objective To explore and study the effect of bundles on prevention and treatment of oral mucositis caused by chemoradiotherapy for patients with nasopharyngeal carcinoma. Methods A total of 40 patients who met the inclusion criteria from June 2014 to December 2014 were selected as the control group, who adopted routine nursing measures, 40 patients who met the inclusion criteria from January 2015 to June 2015 were assigned to the observation group. Bundles on prevention and treatment of oral mucositis caused by chemoradiotherapy for patients with nasopharyngeal carcinoma were edited using a series of evidence-based approach, and it was used to manage the patients of observation group. Results While doing 21, 28, 33 friction of radiotherapy, the oral mucositis level of 0 degree, Ⅰ degree,Ⅱ degree,Ⅲ degree and Ⅳ degree of the observation group were 8, 25, 7, 0, 0 cases;3, 11, 24, 2, 0 cases;0, 19, 13, 6, 2 cases respectively, which were lower than the control group whose degrees were 0, 31, 6, 3, 0 cases;0, 18, 11, 10, 1 cases;0, 9, 17, 9, 5 cases. The difference between the two groups was statistically significant (Z=-4.440,-3.441,-2.232, all P < 0.05 or 0.01). While doing 21, 28, 33 friction of adiotherapy, the throat pain level of 0 degree, Ⅰ degree,Ⅱ degree,Ⅲ degree of observation group were 4, 31, 5, 0; 2, 22, 14, 2; 0, 26, 12, 2 cases respectively, which were lower than the control group whose degrees were 1, 22, 16, 1 cases; 0, 10, 23, 7 cases; 0, 10, 17, 13 cases. The difference between the two groups was statistically significant (Z=-3.137,-3.326,-3.518, all P<0.01). While doing 28, 33 friction of radiotherapy, the Self Rating Anxiety Scale of the observation group scored 56.76 ± 3.19, 58.72 ± 5.41, which were lower than 60.58 ± 2.46, 63.42 ± 4.97 in the control group. The difference between the two groups was statistically significant (t=11.746, 10.561, all P <0.01). While doing 33 friction of radiotherapy, the self rating anxiety scale of the observation group was 60.56 ± 3.73, which was lower than 63.43 ± 4.77 in the control group. The difference between the two groups was statistically significant (t=-4.983, P<0.01). The following entries:swallow, sensation, eating in public, dry mouth, sticky saliva, feel sick of the quality of life questionnaire of the observation group were higher than the control group while doing 33 friction of radiotherapy. All the differences were statistically significant (t=-3.873-5.130, P<0.05 or 0.01). Conclusions The bundles could effectively prevent and treat oral mucositis caused by chemoradiotherapy for patients with nasopharyngeal carcinoma. It could release the throat pain, anxiety and depression of the patients, as well as improve the quality of life to some extent.

Purpose To observe the functional expression of calcium sensing receptor ( CaSR) in human gastric cancer SGC-7901 cell line, the effect of CaSR on intracellular calcium, cell proliferation and migration of SCG-7901. Methods The expression and distribu-tion of CaSR were detected by Western blotting and immunofluorescence observation in SGC-7901. The intracellular concentration of free calcium ( [ Ca2+] i ) was determined by confocal laser scanning microscopy. MTT, flow cytometry and scratch test were used to an-alyze the impact of CaSR the proliferation and the migration capabilities of SGC-7901 cell. Results CaSR protein was expressed in SGC-7901. Extracellular calcium or calindol significantly increased the expression of [Ca2+]i, CaSR and E-cadherin;In addition, the migration capabilities were decreased. Conclusion CaSR is expressed in SGC-7901. The activation of CaSR induces the expression of E-cadherin, and decreases migration ability.

Objective To study the effects of recombinant chimeric toxin Dsg3EC_(1-2)PE40 on T and B cells from PV patients. Methods The recombinant protein Dsg3EC_(1-2)PE40 was expressed on BL21TrxB (DE3) cells, then identified and purified. ELISPOT assay was used to detect and quantitate autoantibody-producing B cells in different concentrations of recombinant chimeric toxin, and MTT assay and ~3H-TdR assay to observe the metabolism and proliferation of T cells from PV patients in vitro. Results The purity of expressed protein Dsg3EC_(1-2)PE40 was up to 80%. The number of anti-Dsg3 antibody-producing B cells in PBMC from PV patients decreased by 40% with treatment of Dsg3EC_(1-2)PE40, which was significantly lower (P

Objective To detect the expression and function of cysteinyl leukotriene receptors (CysLTRs)in keratinocytes.Methods Human keratinocytes were isolated from the tissue of foreskin by digestion with dispase Ⅱ and trypsin,and subjected to primary culture.By using confocal laser scanning microscopy and reverse transcriptase PCR,the localization and expression of CysLTRs were studied in kemtinocytes.respectively.Some primarily cultured keratinocytes were pretreated with leukotriene D4 (30 nmoi/L),MK571(300 nmol/L),and BAYu9773 for 5 minutes followed by the detection of intmcellular calcium level using the Ca2+ indicator dye Fura-2/AM as well as cell proliferation bv MTT assay.Results The expressions of CysLTR1 and CysLTR2 were observed in cultured keratinocytes,and they were mainly located on cell membrane,partly in cytoplasm and nuclei.Compared with non.stimulated cells,a significant increase Was noted in the expression of CysLTRs,especially in the nuclei of keratinocytes stimulated by LTD4(P＜0.05),together with an elevation in intracellular calcium level(42.27±3.00 mmol/L,P＜0.01)and acceleration in cell proliferation (P＜0.01).However,both MK571 and BAYu9773 could completely block the effect of LTD4 on intmcellular calcium level and cell proliferation.and there was no significant difference in the blocking effect between MK571 and BAYu9773.Conclusions Functional CysLTRs are expressed in human keratinocytes.and they carl increase the intracellular calcium level in,and cell proliferation of,keratinocytes.

Objective To compare the efficacy and safety of excimer laser 308 nm phototherapy alone and the combination of excimer laser 308 nm and topical application of vitamine D3 alanogue tacalcitol in the treatment of vitiligo. Methods Seventy-eight patients with vitiligo were enrolled in the single-blind clinical trial, treated with excimer laser 308 nm. The lesions were devided into two groups: patients in the experimental group were instructed to use tacalcitol ointment and the control group were applied with placebo ointment. The lesions were evaluated once per month and photos taken for analyses of clinical effects. Results The results in different locations were compared, the effective rates of the experimental group in cephalofacial site, trunk and limbs were 93.51%, 84.16 % and 42.35 %, respectively. The effective rates of control group in opposite and adjacent sites were 90.9 %, 77.45 % and 34.15 %, respectively (P ＜ 0.05). The comparison of results in different types of lesions indicated that the effective rate of the experimental group in vitiligo vulgaris and segmental vitiligo were 73.81% and 84.00 %, respectively. The effective rate of control group in vulgaris and segmental vitiligo were 86.8 %, 77.45 % and 34.15 %, respectively (P ＜0.05 ). The comparison of results in radiation times and doses of phototherapy showed that the radiation time and dose on the time of initial pigment regeneration were (16. 15 ± 3.22)times and (4.40 ± 5.03)J/cm2 in the experimental group, while ( 18.56 ± 3.50) times and ( 6.60 ± 1.01 ) J/cm2 ( P ＜ 0.05 ) in the control group, the time and dose on the time of apparent effect were ( 20. 36 ± 1.50 ) times and ( 7.50 ± 3.54 ) J/cm2 in the experimental group, and (21.68 ± 2.40) times and( 8.80 ± 9.24)J/cm2 (P ＜ 0.05 ) in the control group. Conclusion Application of tacalcitol ointment in combination with twice-weekly 308 nm excimer laser light phototherapy is an effective alternative treatment for patients with generalized vitiligo.

AIM: To study the effects of fibroblast growth factor 8 (FGF8) on directional differentiation of human dental pulp stem cells (hDPSCs) into odontoblasts and pulp tissue.METHODS: hDPSCs were isolated and cultured, and identified with flow cytometry by detecting cell surface markers of hDPSCs.FGF8 at concentration of 50 μg/L was added into the mineralization fluid to induce the differentiation of the hDPSCs.The mRNA expression of dentin sialophosphoprotein (DSPP), alkaline phosphatase (ALP), bone sialoprotein (BSP) and core-binding factor alpha 1 (Cbfa-1) in differentiated cells was detected by real-time PCR.FGF8 and mouse E11.5 dental epithelium formed restructuring cell group with hDPSCs, and then the restructuring cell group was transplanted under renal capsule membrane in nude mice for tissue culture.DNA in situ hybridization was used to identify the sources of odontoblasts and pulp cells.RESULTS: The surface markers of CD29 and CD90 showed positive in isolated hDPSCs.FGF8 induced hDPSCs to form a distinct mineralization nodule, and the expression of dentin-specific proteins, DSPP, BSP and Cbfa-1, was increased.hDPSCs were induced to differentiate into odontoblasts and pulp cells by E11.5 dental epithelium and FGF8.CONCLUSION: FGF8 can assist dental epithelium to induce directional differetiation of hDPSCs into odontoblasts and pulp cells, and formation of dentin and dental pulp cavity structure.

Objective To evaluate the efficacy and safety of olopatadine hydrochloride for the treatment of chronic idiopathic urticaria (CIU). Methods A multicentre, double-blind, randomized, parallel-group, controlled clinical trial was conducted. A total of 144 patients with CIU from 3 research centers were enrolled into this study, and randomly and equally divided into a test group and a control group. The test group administrated olopatadine hydrochloride 5 mg twice a day for 28 consecutive days, while the control group administrated levocetirizine hydrochloride 5 mg in the forenoon and a placebo tablet of olopatadine hydrochloride 5 mg in the afternoon for 28 consecutive days. The symptom score reducing index(SSRI)served as the primary outcome, and global assessment score for efficacy and total response rates as the secondary outcome. Results Totally, 137 patients completed the trial, including 70 in the test group and 67 in the control group. As intention-to-treat analysis showed, there were no significant differences in the total response rate between the test group and control group on day 7 (64.29% (45/70)vs. 56.72%(38/67), P > 0.05), 14(82.86%(58/70)vs. 74.63%(50/67), P > 0.05), or 28(87.14%(61/70)vs. 77.61%(52/67), P >0.05)after start of treatment. The SSRI was significantly higher in the test group than in the control group after 4 weeks of treatment(82.67% ± 22.70% vs. 70.51% ± 32.07%, P < 0.05). In addition, no significant difference was observed in the incidence of adverse reactions between the test group and control group(33.80%(24/71)vs. 27.94%(19/68), P > 0.05), and adverse reactions mainly included lethargy, dry mouth, fatigue, etc. Conclusion Olopatadine hydrochloride is effective and safe for the treatment of CIU.

OBJECTIVE: To determine the efficacy of 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) in the detection of radiation-induced myocardial damage in beagles by comparing two pre-scan preparation protocols as well as to determine the correlation between abnormal myocardial FDG uptake and pathological findings. MATERIALS AND METHODS: The anterior myocardium of 12 beagles received radiotherapy locally with a single X-ray dose of 20 Gy. 18F-FDG cardiac PET/CT was performed at baseline and 3 months after radiation. Twelve beagles underwent two protocols before PET/CT: 12 hours of fasting (12H-F), 12H-F followed by a high-fat diet (F-HFD). Regions of interest were drawn on the irradiation and the non-irradiation fields to obtain their maximal standardized uptake values (SUVmax). Then the ratio of the SUV of the irradiation to the non-irradiation fields (INR) was computed. Histopathological changes were identified by light and electron microscopy. RESULTS: Using the 12H-F protocol, the average INRs were 1.18 +/- 0.10 and 1.41 +/- 0.18 before and after irradiation, respectively (p = 0.021). Using the F-HFD protocol, the average INRs were 0.99 +/- 0.15 and 2.54 +/- 0.43, respectively (p < 0.001). High FDG uptake in irradiation field was detected in 33.3% (4/12) of 12H-F protocol and 83.3% (10/12) of F-HFD protocol in visual analysis, respectively (p = 0.031). The pathology of the irradiated myocardium showed obvious perivascular fibrosis and changes in mitochondrial vacuoles. CONCLUSION: High FDG uptake in an irradiated field may be related with radiation-induced myocardial damage resulting from microvascular damage and mitochondrial injury. An F-HFD preparation protocol used before obtaining PET/CT can improve the sensitivity of the detection of cardiotoxicity associated with radiotherapy.
Animals ; Dogs ; Fasting ; Fluorodeoxyglucose F18/*metabolism ; Heart/*radiography ; Heart Injuries/*radiography ; Male ; Myocardium/metabolism/pathology ; Positron-Emission Tomography/*methods ; Radiation Injuries/diagnosis/*radiography ; Thoracic Neoplasms/radiotherapy ; Tomography, X-Ray Computed/*methods