Objective In this study,we investigate the influence on nurses professional ability by the hierarchical level training under the advanced mode.Methods By referring to advanced mode of clinical expertise,combined with impacts including the ability of nurses,seniority factors,professional titles and educational background,1 527 nurses of our hospital were divided into five ranks (including nine grades).We developed training plan,training forms and training purposes aiming at all level of training objectives.In addition,we developed a training effect questionnaire and promotion criteria.All the nurses must be evaluated and compared with the traditional nursing mode,which contained 1 486 nurses as the control.Results Within two years after the start of the experiment in the observation group,1 106 nurses had been promoted.Compared with the traditional platform-based training model,nursing assessment indicators were significantly improved by the hierarchical level of training.Quality control of care services was significantly improved and the score of patient satisfaction was significantly increased.Nurses acceptance of tiered training was improved compared to the control group,published research papers were significantly increased.Conclusions Detailed training on the layer classification of the nurses can improve the ability of nurses including specialist knowledge,interpersonal communication and humanistic care,teaching,scientific research,management and other professional skills,so as to improve nursing quality and work efficiency.

Objective To investigate the feasibility of using clustered regularly interspaced short palindromic repeats ( CRISPR)-mediated genome editing to downregulate the expression of programmed cell death protein 1 (PD-1) on primary T cells by using a lentivirus delivery system. Methods Lentivirus vec-tors pLentiCRISPR A1-A6 containing different PD-1 genomic DNA sequences as single guide RNA ( sgRNA) for Cas9 targeting were constructed individually. The lentivirus vectors were tranduced into primary CD4 T cells. Flow cytometry analysis was performed to detect the expression of PD-1 for evaluating the knockout ef-ficiency. Results The lentivirus vectors pLentiCRISPR A1-A6 carrying six different target sites were con-structed and respectively tranduced into primary CD4 T cells. The expression of PD-1 accompanied with the activation of T cells. Co-expression of CD25 and PD-1 was observed on activated T cells. All of the six sites could be targeted by Cas9, of which A2 and A6 sites were more efficient in knocking out the gene encoding PD-1 with a rate of 19% and 29%, respectively. Conclusion This study suggests that it is feasible to knock out the expression of PD-1 on primary T cells by using CRISPR.

Objective To analyze the clinical and pathological characteristics of thyroid nodules.Methods A retrospective analysis of the clinical data of 2843 Han patients with thyroid nodules from January 2009 to June 2011 in Henan province was performed.Results The most high-risk of thyroid nodule resided in the group aged 45-54.The main pathological manifestation was simple nodular goiter.The rate of preoperative misdiagnosis was 2.4％.22.1％ of the patients with thyroid nodules showed abnormal thyroid function,which was mainly as subclinical thyroid dysfunction.The positive rate of thyroid peroxidase antibody (TPOAb) in patients with thyroid nodule was 19.8％ while that of thyroid globulin antibody (TgAb) was 20.0％.Among all 2843 subjects,the prevalence of differentiated thyroid carcinoma (DTC) was 28.8％,and the incidence of DTC was accompanied by raised TSH levels.The prevalence of DTC exceeded 40％ in patients aged between 25 to 34 as well as more than 74 years old.The prevalence of DTC was up to 33.8％ in TgAb positive patients.There was no difference in TPOAb levels between DTC patients and non DTC patients (P =0.406),but the TgAb level in patients with DTC was significantly higher than that of non DTC patients (P =0.033).Conclusion Thyroid nodule patients aged 25-34 or older than 74 formed high-risk groups of DTC.The serum TSH level and TgAb level were positively correlated with the risk of DTC.

Objective:To compare the properties of three kinds of denture base materials treated and untreated by hydraulic pressure cooker. Methods: The impact strength, flexural strength, flexural elastic modulus and color difference roughness of three denture base materials, Kunshan Rijin(KR),GC UNIFAST Trad(GC), Shanghai Zhangjiang(SZ), were measured in vitro. All results were statistically analyzed with SPSS software to have ANOVA and Tamhane's T2 tests, α=0.05. Results: Comparison of the same materials between the treated materials and the untreated materials by hydraulic pressure cooker: there were significant statistical differences of impact strength for all the materials. There were significant statistical differences of color difference for KR and SZ. Comparison of the three materials: impact strength, GC>SZ>KR;flexural strength, KR>SZ>GC;flexural elastic modulus, KR>SZ>GC;roughness, GC>KR>SZ. Conclusion: The properties of denture base materials treated by hydraulic pressure cooker are good.

Objective To estimate the relationship between matrix attachment region-binding proteins (MARBPs) and skin squamous cell carcinoma (SCC), and to screen for highly-associated MARBPs. Methods Skin specimens were resected from the tumor and tumor-adjacent normal tissue of 12 patients with SCC. Normal skin tissue from the abdomen of 2 of the 12 patients served as the control. Reverse-transcription PCR was performed to detect the mRNA expressions of seven MARBPs, including CUX1, Lamin Bl, Nucleolin, hnRNP U,SAFB2, SATB1 and YBX1, in these specimens and in a human keratinocyte cell line HaCaT and two cell lines Colo-16 and A431 derived from human SCC. Results No evident expression was observed for the seven MARBPs in the normal control specimens, while the MARBPs were expressed in the tumor tissue and tumoradjacent tissue specimens, Colo-16 cells and A431 cells at different degrees and frequencies. In HaCaT cells,all the 7 MARBPs except SATB1 were expressed. Statistical analysis showed a significant difference in the expression of CUX1, Nucleolin and YBX1, but not in that of Lamin Bl, hnRNP U, SAFB2 or SATB1 between the tumor and tumor-adjacent normal tissues. Conclusion Some MARBPs such as CUX1, Nucleolin and YBX1 may participate in the development of skin squamous cell carcinoma.

Objective To study the effect of 1,25-dihydroxyvitamin D3 on bone metabolism in diabetic rats and the related molecular mechanism.Methods A total of 45 healthy 6-8 weeks old male Sprague Dawley (SD) rats were treated with streptozotocin.The streptozotocin-induced diabetic rats were randomly assigned to diabetic group (DM),low dose vitamin D treated group(LD),and high dose vitamin D treated group(HD).Another 12 healthy SD rats were used as normol control group(NC).The rats in NC group and DM group were fed with 0.05 ml/d nut oil;those in the LD group and HD group were fed respectively with 0.03 and0.10 μg · kg-1 · d-1 1,25-dihydroxyvitamin D3 dissolved in 0.05 ml nut oil.12 weeks later,serum calcium,phosphorus,osteocalcin,type Ⅰ collagen cross-linked telopeptide (NTX),and 24 h urinary calcium were determined.Right femurs were harvested for pathohistological analysis by hematoxylin-eosin (HE) staining.Expressions of osteoprotegerin,receptor activator nuclear factor κB ligand (RANKL),core binding factor α1(Cbfa1) were detected by immunohistochemical staining.The osteoprotegerin,RANKL,Cbfa1,osteocalcin mRNA levels of bone tissue were performed by realtime quantitative reverse transcription polymerase chain reaction assay.Results (1) Compared with the NC group,serum calcium and 24 h urinary calcium in LD and HD groups were significantly higher (P＜0.05) ; 24 h urinary calcium in DM group was significantly higher than that in NC group (P ＜ 0.05).(2) Serum osteocalcin level in DM and LD groups was significantly lower than that in NC group (P＜0.05) ; there was no significant difference between the serum NTX levels of all groups (P＞0.05).(3) There was no significant difference in the mRNA expression of Cbfa1 in all groups (P＞0.05).The mRNA expression of osteocalcin in DM group was significantly lower than that in NC group (P ＜0.05).The mRNA expression of RANKL in DM group was significantly lower than that in NC group (P＜0.05) ; and that in LD and HD group was significantly higher than that in DM group (P＜0.05),and that in HD group was significantly higher than that in LD group (P＜0.05).The mRNA expression of osteoprotegerin in DM group was significantly lower than that in NC group (P＜0.05).The ratio of RANKL to osteoprotegerin in HD group was significantly higher than that in DM group (P＜0.05).Conclusions Vitamin D may promote bone metabolism in diabetic rats by up-regulating the expressions of osteocalcin and RANKL or in addition to other means.

Objective To investigate cleanliness of hospital environmental object surfaces and hands of health care workers(HCWs).Methods The adenosine triphosphate (ATP)bioluminescence assay was used to detect object surfaces and hands of HCWs in a hospital,on-the-spot intervention was conducted.Results The qualified rates of hospital environmental object surfaces and ventilator-relevant object surfaces were 58.14% (200/344)and 69.88%(116/166)respectively,the qualified rate of ventilator tracheal intubation site was low (29.17%);the qualified rate of telephone surfaces was the lowest (27.27%).The qualified rates of ventilator-relevant object surfaces used con-tinuously for ≥48h and <48 h were 56.70%(55/97)and 88.41 %(61/69)respectively,there was significant differ-ence between the two(χ2 =19.26,P <0.01).The qualified rates of HCWs’hands before and after intervention were 34.18% and 85.58% respectively,relative light unit (RLU)values were (1 033.46±106.20)and (80.46±10.68) respectively,the qualified rates and RLU before and after intervention were both significantly different (both P <0.01).Conclusion Contamination of object surfaces and hands’of HCWs in hospital dynamic environment is seri-ous,ATP bioluminescence detection and on-the-spot intervention is helpful for improving cleanliness of hospital en-vironment object surfaces and HCWs’compliance to hand hygiene.

Objective To investigate the factors affecting the vaginal birth after cesarean (VBAC). Methods Totaly 298 women who underwent trial of labor after cesarean section (TOLAC) from Jan 2015 to Dec 2015 were recruited from Beijing Obstetrics and Gynecology Hospital, FuXing Hospital, Tongzhou Maternal and Child Health Hospital of Beijing, the Second Affiliated Hospital of Chongqing Medical University and the People′s Hospital of Chengyang District of Qingdao. The maternal age, the interval from the last cesarean section, the body mass index (BMI) before pregnancy, the weight gain during pregnancy, the way into labor, the Bishop score before labor, the gestational age and the birth weight of the neonate were recorded in a self-made form. The factors affecting VBAC were analyzed by univariate analysis and multivariable logistic regression. Results (1)The incidence of VBAC, uterine rupture, postpartum hemorrhage and neonatal asphyxia were 70.5%(210/298), 2.7%(8/298), 9.4% (28/298) and 1.3% (4/298), respectively. No maternal death and perinatal death occurred. (2)The univariate analysis suggested that the maternal age, the BMI before pregnancy, the Bishop score before labor, the labor induction, the gestational age at delivery and the neonatal weight were factors affecting VBAC. The maternal age and the Bishop score before labor were significantly higher in the VBAC group than in the unsuccessful TOLAC group(P<0.05). While the BMI before pregnancy, the induction rate, the gestational weeks at delivery and the birth weight of the neonate were significantly lower in the VBAC group than in the unsuccessful TOLAC group (P<0.05). Multivariable logistic regression analysis showed that successful VBAC was affected by the maternal age, the BMI before pregnancy, the Bishop score before labor and the birth weight of the neonates(P<0.05). Conclusion The maternal age, the BMI before pregnancy, the Bishop score before labor and the birth weight of neonate are the main factors affecting VBAC.

Aim To observe the variation of T-cell subsets in spleen in different courses of disease and tis-sue histopathological changes in rats with adjuvant ar-thritis (AA).Methods The model of rat AA was in-duced by complete Freund’s adjuvant into the right hind metatarsal footpad of 20 male Lewis rats.Arthritis was evaluated by total score,arthritis index and paw swelling number. The percentage of total (CD3 +CD4 +), memory (CD4 + CD44 +), no-sensitizated (CD4 + CD62L +) and Th1 7 (CD4 + IL-1 7 +) on CD4 +T cells in spleen were detected by flow cytometry on days 1 4 and 22 after immuniozation.The his-topathological evaluation of heart,liver,spleen,lung, kidney,ankle joints and mesenteric lymph nodes were examined by hematoxylin and eosin (HE)stain.Re-sults The onset of secondary arthritis appeared on a-bout day 1 0 after immunization,with a peak onset on day 1 9.Systematic arthritis symptoms included front and hind paw swelling and redness (swelling was more apparent in the front rather than hind paws),nodule formation and redness on the ears,connective tissue swelling and redness of the nose and evident nodules and redness on the tail.Total score,arthritis index and the paw swelling numbers were enhanced in AA rats. Pathohistology of ankles joints showed hyperplasia, pannus formation,and inflammatory cells infiltration in AA rats.And pathohistology of heart,liver,spleen, kidney and mesenteric lymph nodes showed inflamma-tory changes.Compared with normal group,there was no change in the percentage of total,memory and no-sensitizated CD4 +T cell in the early stage inflammation but they were significantly enhanced in the peak of in-flammation.Th1 7 cell subsets were significantly en-hanced in both of the early and peak of inflammation. Conclusion These findings suggest that abnormal T cell responses and tissue histopathological changes are important features of AA rats.

[ ABSTRACT] AIM:To investigate the effect of polysaccharide from Fructus corni ( PFC) on cardiomyocytes against hypoxia/reoxygenation ( H/R) injury and its possible relationship with ROS/PKC/p38 MAPK pathway.METHODS:Prima-ry cardiomyocytes were isolated from neonatal SD rats and randomly divided into normal group, H/R group, PFC (20 mg/L, 100 mg/L and 200 mg/L) preconditioning+H/R groups, chelerythrine+PFC (100 mg/L)+H/R group and SB203580+PFC (100 mg/L)+H/R group.The cell viability was measured by inverted microscopic observation.Apoptosis in the car-diomyocytes was detected by Hoechst 33258 staining and fluorescence microscopy.The levels of lactate dehydrogenase ( LDH) and superoxide dismutase ( SOD) in the cell culture supernatants, and the reactive oxygen species ( ROS) in the cells were also measured by microplate reader.The protein levels of PKC, p-p38 MAPK and HSP70 in the cells were detec-ted by Western blotting.RESULTS:Compared with normal group, the cell viability and beating frequency were decreased in H/R group.LDH and ROS contents, apoptotic rate and p-p38 MAPK level increased significantly (P<0.01).Compared with H/R group, PFC preconditioning increased beating frequency, SOD activity and the protein level of PKC and HSP70, and decreased ROS production, the protein level of p-p38 MAPK and cell apoptotic rate.However, the effect of PFC was in-hibited by chelerythrine or SB203580.CONCLUSION:PFC may protect cardiomyocytes from hypoxia/reoxygenation injury. Its mechanism is possibly involved in the inhibition of ROS via increasing the activity of SOD and the activation of PKC, and suppression of excessive activation of p38 MAPK.