Objective To investigate the correlation of epidermal distribution of lamellar bodies and expression of ceramidase with skin barrier dysfunction in polymorphous light eruption.Methods Forty-seven patients with polymorphous light eruption and 40 healthy volunteers were recruited into this study.Noninvasive instruments were used to measure skin sebum content,transepidermal water loss(TEWL)and water content in stratum corneum in all of the subjects.Then,tissue specimens were obtained from the lesions at sunexposed sites in the patients and normal skin of the healthy volunteers.The ultrastructure and distribution of lamellar bodies were observed with transmission electron microscopy in five lesion and control specimens.Immunohistochemistry was performed to detect the expression of ceramidase in the tissue specimens.Results Compared with the normal skin from healthy volunteers,the lesions from patients showed decreased number of lamellar bodies in the granular layer and prick cell layer with a disorganized arrangement.Ceramidase was positively expressed in 20 lesion specimens and 36 normal control specimens,weakly expressed in 21 lesion specimens and 4 normal control specimens,and negative in 6 lesion specimens; there was a significant difference in the expression of ceramidase between the lesion specimens and normal control specimens(P ＜ 0.01).The lesions also showed high TEWL(34.2191 ± 12.70 vs.16.8350 ± 6.50,P ＜ 0.01),lower water content in stratum corneum(22.7319 ± 8.71 vs.29.4250 ± 5.08,P ＜ 0.01)and similar skin sebum content compared with the normal skin.Conclusions There is a disturbance in the synthesis of ceramide in patients with polymorphous light eruption,which may contribute to the impairment of skin barrier.

Objective To investigate the antimicrobial resistance proifle of Shigella flexneri in Shanghai from 2010 to 2014 and examine the mechanism of fluoroquinolone resistance.Methods Kirby-Bauer method was used to determine the susceptibility of the S. flexneri strains to 14 antibiotics. The minimum inhibitory concentration (MIC) of ciprofloxacin was tested by E-test. Mutations within quinolone resistance determining regions (QRDR) of gyrA and parC and plasmid-mediated quinolone resistance (PMQR) genes,qnrA,qnrB,qnrS andaac(6')-Ib-crwere identified by polymerase chain reaction (PCR). All the products were subjected to sequencing analysis.Results More than 90 % of the 139S. flexneri isolates were resistant to ampicillin, streptomycin, tetracycline, and nalidixic acid, 40.3 % to ciprofloxacin and 30.2 % to cefepime, respectively. Genetic mutation of gyrA and parC was found in 98.6 % and 97.8 % of the strains, respectively. Three point mutations (Ser83, Asp87 and His211) were detected in gene gyrA and one point mutation (Ser80) was found in in gene parC. Plasmid-mediated resistant gene qnrS was found in 9 strains and aac(6')-Ib-cr in 6 strains.Conclusions The antibiotic resistance of S. flexneri is serious in Shanghai. The mutation rate within QRDR is high. Point mutation in Asp87 of gyrA is the main mechanism of quinolone resistance. The plasmid-mediated quinolone-resistant genes also play an important supplementary role.

Objective To identify and compare melanin distribution in iris and skin tissues of rabbits from four breeds including WHBE rabbit.Methods Ferrous sulphate staining was performed for iris and skin tissue of rabbits from different breeds.Results It showed dark green staining in both iris stroma and iris pigment epithelium of WHBE rabbit, while light green staining was observed in its follicles.Conclusion The results of histologic evaluations provide some clues to the future research on mechanism of WHBE rabbit phenotype mutation.

Objective:To evaluate the effects of wearing N95 mask on the quality of chest compression and fatigue.Methods:A total of 80 participants from Zhongnan Hospital with basic life support (BLS) certification conferred by American Heart Association (AHA) within two years were enrolled. After reviewing the key points of cardiopulmonary resuscitation (CPR) and grasping the operation on the manikin, they were randomized (random number) into two groups: wearing surgical masks (SM group, n=40) and wearing N95 masks (N95 group, n=40) during CPR. Each participant performed a 2-minute chest compression-only CPR on the manikin. Participants' height, body weight, Borg scores and physiological parameters before and after CPR were recorded. The quality of chest compression (including compression depth, compression rate, adequate depth proportions, adequate rate proportions, hand position and complete chest recoil) were recorded by Laerdal QCPR ? software. Student's t test and Mann-Whitney test were used to compare the differences of chest compression quality indexes between the two groups. Results:Compared with the SM group, participants in the N95 group had significantly increased median of Borg scores after CPR (16 vs 14, P=0.027), and decreased quality of chest compression, including the decline in compression depth (mean 47 mm vs 52 mm, P=0.020), compression rate (107 times/min vs 118 times/min, P=0.004), complete chest recoil rate (89.8% vs 98.1%, P=0.046), adequate depth proportions (67.4% vs 89.6%, P<0.01) and adequate rate proportions (60.6% vs 74.8%, P<0.01). Conclusions:Wearing N95 masks during CPR decreases the quality of chest compression and aggravates rescuers’ fatigue. Therefore, it is necessary to exchange rescuers more frequently to ensure the quality of chest compression when wearing N95 masks.

ObjectiveTo explore the effects of epidermal proteins and lamellar bodies on skin barrier in glucocorticoid-dependent dermatitis.MethodsTotally,60 patients with glucocorticoid-dependent dermatitis and 40 normal human controls were eligible for this study.A noninvasive method using TewameterTM was applied to determine transepidermal water loss (TEWL) value in these subjects.Tissue specimens were obtained from the lesions of 13 patients with glucocorticoid-dependent dermatitis and normal skin of 10 human controls.Subsequently,haematoxylin and eosin(HE) staining was performed to observe the histopathological changes,immunohistochemistry to detect the protein expressions of K6,K10,K14,K15,loricrin,filaggrin,involucrin in epidermis,and electron microscopy(EM) to estimate the density of lamellar bodies in tissue specimens.ResultsCompared with the normal controls,the patients displayed an elevated TEWL value (P ＜ 0.05),which suggested an impaired epidermal barrier.Histopathology of lesions revealed nonspecific inflammatorychanges withmarkeddifferencesbetweendifferentclinicaltypesofglucocorticoid-dependentdermatitis.Immunohistochemistry revealed an attenuated expression of K10,K14,loricrin,filaggrin,involucrin and abnormal expression of K15 in lesional epidermis compared with the normal epidermis (all P ＜ 0.05),hinting a suppression of epidermal differentiation and proliferation as well as an impairment of cornified envelope structure.The number and density of lamellar bodies were also reduced in lesional epidermis compared with the control epidermis.ConclusionsCompared with normal skin,the structure of skin barrier is impaired in lesions of glucocorticoid-dependent dermatitis,to restore skin barrier is essential for the treatment of this entity.

An HPLC pre-column derivatization detection method was established to detect and analyze the formaldehyde and acetaldehyde in polysorbate 80 and polysorbate 20 from different manufacturers.The effects of aldehyde and acetaldehyde on the aggregation of adalimumab under different conditions were monitored.Based on the control of genotoxic impurities and the influence on the stability of monoclonal antibody preparations, the control limits of the two chemicals were preliminarily obtained.2, 4-dinitrophenylhydrazine (2, 4-DNPH) was applied as the derivatization reagent in HPLC pre-column derivatization; acetonitrile and water were used as mobile phase to perform a gradient elution on a C8 (4.6 mm × 150 mm, 5 μm) chromatographic column.The detection wavelength was 360 nm, and the external standard method was used for quantification.Verification results showed that the method was suitable for the quantitative analysis of trace formaldehyde and acetaldehyde in polysorbate 80 and polysorbate 20 . The detection and analysis of formaldehyde or acetaldehyde in different batches of polysorbate 80 and polysorbate 20 from different manufacturers showed that the content of formaldehyde and acetaldehyde were quite different. The content of formaldehyde and acetaldehyde in polysorbate 80 were significantly higher than those of polysorbate 20. After monitoring the changes of adalimumab aggregates treated by formaldehyde and acetaldehyde by size exclusion chromatography (SEC), it was found that the effect of formaldehyde on adalimumab aggregation was significantly higher than that of acetaldehyde.According to the requirements of ICH M7 (International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use, M7: Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk), the impurity limits of formaldehyde and acetaldehyde in polysorbate 80 and polysorbate 20 for monoclonal antibody preparations were calculated from the perspective of risk assessment.Combined with the influence on the aggregation stability of monoclonal antibodies, the preliminary limis for acetaldehyde and acetaldehyde were recommended to be ≤ 7 μg/g and ≤ 765 μg/g, respectively.

Objective To observe the effect of ultraviolet irradiation comparising 95％ ultraviolet A (UVA)and 5％ ultraviolet B(UVB)on the proliferation of human epidermal keratinocytes(HEKs),in hope to offer a basis for the construction of a photodamaged skin model induced by sunlight.Methods HEKs were isolated from foreskin tissue and cultured in vitro.After several passages,the HEKs were irradiated with different doses(0,2.5,5,10,20,30,40,60 J/cm2)of UV comprising 95％ UVA and 5％ UVB.Methyl thiazolyl tetrazolium(MTT)assay was used to evaluate cell viability after 24 hours of additional culture.SPSS 17.0 software was used to calculate the median lethal dose(LD50)of ultraviolet radiation in HEKs.Results The proliferation of HEKs was inhibited by 0,1.03％,6.60％,17.28％,31.28％,49.59％,59.67％ and 70.99％ respectively after irradiation with UV of 0,2.5,5,10,20,30,40 and 60 J/cm2.A significant inhibition of cell proliferation was observed in HEKs irradiated with UV at a dose of no lower than 10 J/cm2 compared with unirradiated HEKs(F =62.11,P ＜ 0.05).The LD50 of UV in HEKs was 31.31 J/cm2.Conclusions Aas the dose of UV irradiation increases,the proliferative activity of HEKs decreases,with the LD50 of UV being 31.31 J/cm2.

Objective To analyze the genetic variation among white hair black eyes (WHBE) rabbit, Japanese white ( JW) rabbit and New Zealand white ( NZW) rabbit using random amplified polymorphic DNA ( RAPD) technique . Methods Thirty rabbits (male/female 1∶1) of each strain were used in this study.The genomic DNA was extracted from 90 rabbits.Sixty arbitrary primers were used to amplify DNA of rabbits with RAPD-PCR method.Based on the preliminary experiments , polymorphic primers were selected to analyze the genetic variation among the three rabbit strains .The experi-mental data were analyzed using Popgene 3.2 software.Results (1) Twenty-five polymorphic primers were selected among 60 arbitrary primers.493 amplified fragments were detected ranging from 100 bp to 1800 bp.Sixteen primers among 25 arbitrary primers could not only amplify the common DNA bands of 3 rabbit breeds , but also amplify particular alleles in the WHBE rabbit.(2) 234 RAPD sites were detected by agarose gel electrophoresis in WHBE rabbit , among which 166 sites were polymorphic , accounting for 70.94%.228 RAPD sites were detected by agarose gel electrophoresis in the JW rabbit, while 122 sites of them were polymorphic , accounting for 53.51%.231 RAPD sites were detected by agarose gel e-lectrophoresis in the NZW rabbits , with 94 sites being polymorphic, accounting for 40.69%.(3) The Shannon genetic di-versity index of WHBE rabbit, JW rabbit and NZW rabbit was 0.3385, 0.2222 and 0.1905, respectively.(4) The genet-ic similarity between JW rabbit and NZW rabbit was highest among the three rabbit breeds (0.8443), followed by that be-tween WHBE rabbit and JW rabbit (0.8204), and the genetic similarity between WHBE rabbit and NZW rabbit (0.7862) was the lowest .Conclusions Our results demonstrate that there are both genetic similarities and genetic variations among WHBE rabbit, JW rabbit and NZW rabbit .The RAPD technique can be used to delect the genetic relationships among dif-ferent breeds and different individuals of the same breed of rabbits .

AIM: To investigate the autophagy of breast cancer cells induced by baicalein and to explore its mechanism.METHODS: The effects of baicalein on the viability of MCF-7 cells and 4T1 cells were investigated by MTT assay, and the dosage of the drug was determined.The expression levels of microtubule-associated protein 1 light chain 3-II (LC3-II) and LC3-I in the MCF-7 cells and 4T1 cells treated with baicalein at doses of 25, 50 and 100 μmol/L, or combined with autophagy inhibitor 3-methyladenine (3-MA) were determined by Western blot.In order to confirm the role of baicalein in autophagy, the effect of 3-MA on the apoptosis of both MCF-7 cells and 4T1 cells induced by baicalein was analyzed by flow cytometry.The protein levels of p-mTOR, mTOR, p-AKT and AKT were examined by Western blot and the role of AKT-mTOR pathway in the induction of autophagy in breast cancer induced by baicalein was determined by the combination of activators.RESULTS: Baicalein at 50 μmol/L and above doses significantly inhibited the viability of breast cancer cells in a dose-and time-dependent manner.The expression of LC3-II/LC3-I in both MCF-7 cells and 4T1 cells was significantly enhanced after the action of baicalein, and the ratio of LC3-II/LC3-I was significantly decreased after 3-MA addition.The results of flow cytometry showed that, compared with baicalein group, the combination of baicalein and 3-MA promoted the levels of necrosis and apoptosis.Moreover, the protein levels of p-mTOR and p-AKT were significantly decreased and were rescued by EGF, while their total protein levels were not changed.CONCLUSION: Baicalein induces autophagy through AKT-mTOR pathway both in MCF-7 cells and 4T1 cells.

Objective To investigate the impact of aging and vascular endothelial function on arterial stiff- ness in patients with isolated systolic hypertension.Methods Patients with isolated systolic hypertension (ISH,n=75)age-matched healthy subjects(n=30)and young healthy subjects(n=50)were submitted to deter- mination of aortic pulse wave velocity(baPWV)and vascular endothelial function evaluated by flow-mediated dila- tion(FMD).Results baPWV was progresively decreased(ISH:2459.2?436.8 vs elderly healthy:2097.2? 315.7 vs young healthy:1619.7?214.2 cm/s,P