Cytokeratins can maintain the morphous and function of normal cells. Clinically, cytokera-tins are used to diagnose several tumors as tumor markers. Recently, numerous studies indicate that the abnor-mal expression, mutation in the site of phosphorylation and hyperphosphorylation have relationship with the mechnism of tumorigenesis. Reseach of the relationship is important for the diagnosis, therapy and prognosis of tumor.

Adult ; Chromosome Aberrations ; drug effects ; Ethylene Oxide ; poisoning ; Humans ; Lymphocytes ; drug effects ; pathology ; Male ; Micronuclei, Chromosome-Defective ; drug effects

Objective To evaluate the therapeutic effect of narrow band-ultraviolet B (NB-UVB) alone or in combination with interferon-alpha-2b (INF-alpha-2b) for mycosis fungoides (MF),and to assess the correlation between the therapeutic effect and peripheral blood regulatory T (Treg)/T helper type 17 (Th 17) cells.Methods Thirty-three patients with stage ⅠA to ⅡA MF were randomly divided into two groups:NB-UVB group (n =15) receiving NB-UVB radiation alone,combined group (n =18) treated with NB-UVB radiation and intramuscular injection of INF-alpha-2b.Ten healthy volunteers were selected as the control group.Peripheral blood samples were collected before and 9 months after the start of treatment.Flow cytometry was performed to determine the percentages of Treg cells and Th17 cells.Statistical analysis was carried out by using t test,one-way analysis of variance,and Fisher's exact test.Results The average treatment duration was 9 months among these patients.Therapeutic outcomes were significantly better in the combined group than in the NB-UVB group (P =0.023).Among the 15 patients in the NB-UVB group,6 achieved complete remission,3 partial remission,6 showed no response;of the 18 patients in the combined group,12 experienced complete remission,5 partial remission,and 1 showed no response.Before the treatment,the percentages of both Treg and Th17 cells in peripheral blood were significantly higher in the NB-UVB group and combined group than in the control group (both P ＜ 0.05),but similar between the NB-UVB group and combined group (both P ＞ 0.05).After the treatment,the percentages of both Treg and Th17 cells in the NB-UVB group and combined group significantly decreased compared with those before the treatment,but were still higher than those in the control group (both P ＜ 0.05).Additionally,the degree of decrease in the percentages of Treg and Th17 cells was significantly greater in the combined group than in the NB-UVB group (both P＜ 0.05).The seven patients with no response also showed a significant decrease in the percentage of Treg cells (P ＜ 0.05),but no obvious changes in that of Th 17 cells (P ＞ 0.05) after the treatment.Conclusions The therapeutic effect of NB-UVB radiation combined with intramuscular INF-alpha-2b is superior to that of NB-UVB radiation alone for MF,which may be associated with the degree of decrease in peripheral blood Treg and Th 17 cells.

Objective:To investigate the expression and clinical significance of effector T cell (Teff), regulatory T cell (Treg) and their cytokines in patients with vitiligo.Methods:84 patients with vitiligo (38 cases in stable stage, 46 cases in advanced stage) who were admitted to the Dermatology Department of the Dezhou People's Hospital between June 2018 and June 2019 and 30 healthy people (control group) were enrolled in the study. The levels of Teff subsets and Treg cells in peripheral blood were detected by flow cytometry. The levels of Teff subsets and Treg cells-associated cytokines were detected by enzyme-linked immunosorbent assay (ELISA). The relationship between Teff subsets and Treg cells levels in peripheral blood, relationship between Teff subgroups, Treg cells-associated cytokines and course of vitiligo, skin lesion area were analyzed.Results:The levels of T helper cell (Th)17 and Th22 in peripheral blood of vitiligo group were significantly higher than those of control group, while levels of Treg cells were significantly lower than control group ( P<0.05). Compared with the stable vitiligo group, the levels of Th17 and Th22 in peripheral blood were higher and levels of Treg cells were lower in the advanced vitiligo group ( P<0.05). Treg cell level was negatively correlated with Th17 cell level ( r=-0.303, P=0.001), and negatively correlated with Th22 cell level ( r=-0.200, P=0.033). The levels of serum interleukin (IL)-17 and IL-22 in vitiligo group were significantly higher than those in control group, while the level of transforming growth factor-β (TGF-β) was significantly lower than that in control group ( P<0.05). Compared with the stable vitiligo group, levels of serum IL-17 and IL-22 were higher and level of TGF-β was lower in the advanced vitiligo group ( P<0.05). Correlation analysis showed that levels of IL-17 and IL-22 were positively correlated with skin lesion area ( r=0.361, 0.288, P<0.05), and level of TGF-β was negatively correlated with skin lesion area ( r=-0.312, P<0.05). Conclusions:Teff/Treg cells are unbalanced in patients with vitiligo, showing obvious shift to Teff subsets. The progression of vitiligo may be related to Teff subsets and Treg cells-associated cytokines.

BACKGROUND:YY1 is mainly expressed in the undifferentiated epidermic cells in mouse basal lamina, and the expression level is gradual y down-regulated as the differentiation towards suprabasal lamina. The differential expression indicates that, YY1 is one of the regulators in the process of epidermic cells differentiation.
OBJECTIVE:To observe the effects of YY1 over-expression on the differentiation of HaCaT cells infected with lentivirus.
METHODS:Lentivirus-YY1 was transferred into the HaCaT cells by using Lipofectamine 2000. After selection of the puromycin, monoclonal celllines were established, and the control group were lentivirus-infected HaCaT cells and uninfected HaCaT cells. The expression of YY1 was detected by using western blot analysis. cells in Lentivirus-YY1-HaCaT group and HaCaT-YY1 group were further divided into two subgroups according to the calcium concentration in culture medium, cells were either cultured in low-calcium medium (0.12 mmol/L) for 48 hours, or cultured in low-calcium medium (0.12 mmol/L) for 24 hours and in high-calcium medium (0.35 mmol/L) for additional 24 hours. Keratin K1, K10, K14, and involucrin, filaggrin and loricrin after over-expression of YY1 were detected with western blot analysis.
RESULTS AND CONCLUSION:The HaCaT cells were successful y infected with lentivirus-YY1, and we obtained over-expression of YY1 protein in monoclonal celllines under high-calcium concentrations, the over-expressed YY1 could decrease the expression of K1, involucrin and loricrin, thereby preventing the process of epidermal keratinocytes and maintaining the cells in an undifferentiated state. Lentivirus can efficiently infect human immortalized epidermal cellHaCaT, and YY1 may the important factor of inhibiting the differentiation of basal epidermal cells and maintaining the undifferentiated proliferation status.

ObjectiveTo detect five periodontal pathogenic bacteria in coronary atherosclerotic plaques.MethodsAtherosclerotic plaque specimens were obtained from 101 patients who scheduled for coronary artery bypass graft surgery.The bacteria DNA was obtained from coronary atherosclerotic plaques using the chelex-100 method.The extracted DNA were examined by PCR.ResultsWithin the 101 pieces of atherosclerotic plaque samples Porphyromonas gingivalis( Pg,31％ ),Tannerella forsythensis(Tf,42％ ),Prevotella intermedia( Pi,26％ ),Fusobacterium nucleatum( Fn,21％ ),Actinobacillus actinomycetemcomitans( Aa,23％ ).PCR products were sequenced and were compared with GenBank sequences,the homology was 99％-100％.ConclusionPeriodontitis might affect the development of atherosclerosis and there is a correlation between coronary heart disease and chronic periodontitis.

Objective To investigate the application value of spatio-temporal image correlation (STIC) combined with tomographic ultrasound imaging(TUI) in the prenatal diagnosis of conotruncal defects(CTD).Methods Two-dimensional(2D) fetal echocardiography to screen and TUI-STIC volumes from 1508 cases of fetuses of high risk with congenital heart disease.Postnatal work-up and pathological results were available for all fetuses with CTD.Results Thirty nine cases with CTD were found by TUI-STIC while thirty five cases were found by 2D echocardiography,but TUI-STIC had new findings and corrected the diagnosis in 9 cases as compared with 2D echocardiography.The sensitivity,specificity,positivity predictive value,negative predictive value and accuracy of TUI-STIC in evaluating CTD were 97.5 ％,100％,100 ％,99.9 ％ and 99 ％.The Kappa value of consistency test between 2DE and TUI-SIC was 0.244(P ＜ 0.01),McNemar test showed that the difference was statistically significant (P ＜ 0.01).Conclusions TUI-STIC allows a complete sequential analysis of fetal conotruncal defects and supplying additional information over 2D fetal echocardiography,it could improve the prenatal diagnosis rate.TUI-STIC is helpful in diagnosis of prenatal conotruncal defects.

Objective To investigate the changes of T helper type 17 (Thl7) cells and regulatory T (Treg) cells in different stages of mycosis fungoides.Methods Flow cytometry was performed to determine the percentage of Treg and Th17 cells in peripheral blood from 28 patients with mycosis fungoides (MF),13 patients with large plaque parapsoriasis (PP),17 patients with lichen planus (LP) and 10 healthy human controls,and immunohistochemistry to detect the expressions of forkhead box protein 3 (FOXP3) and interleukin (IL)-17 in tissue specimens from 40 patients with MF,13 with PP,17 with LP and 10 healthy human controls.Statistical analysis was carried out by one-way analysis of variance and Pearson correlation analysis.Results As far as the percentage of Treg cells in peripheral blood was concerned,MF,PP and LP patients were significantly higher than the healthy controls (8.09％ ± 1.68％,6.53％ ± 1.67％ and 2.84 ％ ± 1.16％ vs.1.01％ ± 0.35,all P＜ 0.05),PP patients were higher than LP patients and healthy controls (both P ＜ 0.05),and LP patients were higher than healthy controls (P ＜ 0.05).The percentage of Th17 cells in peripheral blood was significantly increased in MF patients compared with PP patients,LP patients and healthy controls (3.22％ ± 0.82％ vs.2.46％ ± 0.79％,1.38％ ± 0.47％ and 0.59％ ± 0.30％,all P ＜ 0.05).Elevated expression rate of FOXP3 was observed in MF,PP and LP lesions as compared with normal skin (14.94％ ± 4.46％,11.95％ ± 4.72％,6.32％ ± 2.81％ vs.3.43％ ± 1.79％,all P ＜ 0.05),and in MF and PP lesions compared with LP lesions (both P ＜ 0.05),but no significant difference was observed between MF and PP lesions (P ＞ 0.05).There was a significant increase in the expression rate of IL-17 in MF lesions compared with PP lesions,LP lesions and normal skin (15.89％ ± 4.27％ vs.12.02％ ± 3.34％,4.84％ ± 1.93％ and 2.62％ ± 0.89％,all P ＜ 0.05).The Th17/Treg ratio in peripheral blood was significantly lower in MF and PP patients than in LP patients and healthy controls (0.41 ± 0.11 and 0.39 ± 0.12 vs.0.50 ± 0.06 and 0.57 ± 0.19,all P ＜ 0.05).A positive correlation was observed between the proportion of Thl7 cells and Treg cells (r =0.423,P ＜ 0.05) in patients with early-stage MF,but not in those with tumor-stage MF.The proportion of Th17 cells decreased,but that of Treg cells continuously increased in patients with tumor-stage MF.However,no significant difference was noted in the proportion of Thl7 cells or Treg cells among patients with different stages of MF.Conclusion The imbalance between Treg and Th17 cells may be involved in the occurrence and development of MF.

Objective To observe changes of the pancreas after limb ischemia reperfusion, and the protective effect of taurine (Tau) postconditioning. Methods Twenty-four male SD rats were randomly divided into control group (Control), limb ischemia reperfusion (LIR) group, and taurine postconditioning (Tau) group (n=8 for each group). Plasma contents of xanthine oxidase (XOD), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), pancreatic amylase (AMS), pancreatic lipase (LPS) and trypsin were detected in three groups. Morphological changes of the pancreas were observed using optical microscopy. The expressions of anti-C/enhancer-binding protein homologous protein (CHOP) in pancreas were analyzed by immunohistochemistry assay. TUNEL staining was performed to estimate the apoptosis of pancre-atic cells. Results Compared with the control group and Tau group, plasma contents of MDA, XOD, ROS, AMS, LPS, and trypsin were significantly increased in LIR group, but the level of SOD was significantly lower in LIR group ( P<0.05). HE staining showed that acinar structure was disrupted , pancreatic lobule gap widened, stromal vascular dilatation and conges-tion were observed in LIR group. The perivascular infiltration of inflammatory cells appeared, islet cells were damaged with irregular islet. The immunohistochemical analysis showed the increased expression of CHOP in pancreas in LIR group than that of Tau group. And the pancreatic apoptosis was enhanced in LIR group detected by TUNEL staining. Conclusion Tau-rine postconditioning can ameliorate pancreatic injury following limb ischemia reperfusion, which may be related to the inhi-bition of oxidative stress, down-regulation of CHOP expression, thereby reducing endoplasmic reticulum stress-induced apoptosis.

Purpurin is a common component ofRubia cordifolia L. The study on its molecular target was useful for elucidating the therapeutic material basis and action mechanism ofR. cordifolia. HT-29 cells were used in the cell culture. The highly expressed G Protein-coupled Receptor-35 (GPR35) agonist was used as target. The label-free optical biosensor cellular assay was used to investigate the agonist activity ofpurpurin at an endogenous receptor. The results showed thatpurpurin can cause DMR response in HT-29 cells. And the DMR response curve type was consistent with zaprinast. Its EC50 was 6.142± 0.189μmol·L-1. In addition,purpurinhad desensitization effect on GPR35 agonist zaprinast in HT-29 cells. GPR35 agonist ML145 blocked the DMR ofpurpurin. It was concluded thatpurpurinwas the GPR35 agonist.