ObjectiveTo develop a new nested PCR to detect human papillomavirus (HPV) DNA in lesions of extragenital Bowen's disease.Methods DNA was extracted from the lesions of 41 patients with extragenital Bowen's disease and suhjected to the amplification of HPV by a nested PCR.Five primers,including CN1FR,CN2FR,CN3FR,CN4FR and CN5FR,were designed and used in the second round PCR.ResultsBased on the ClustalX analysis,69 HPV subtypes,including mucosal types,cutaneous types and epidermodysplasia verruciformis-associated types,could be amplified by using the 5 designed primers.The detection limit varied from 10-3 to 10-2 copies of HPV DNA for this PCR.Of the 41 lesional specimens,5 were positive for HPV DNA,including 3 cases of high-risk HPV types(2 cases of HPV 16,1 case of HPV 33) and 2 cases of cutaneous HPV types(1 case of HPV 27 and 1 case of HPV 76).ConclusionsThe improved nested PCR is highly sensitive and specific for the detection of HPV DNA in lesions of extragenital Bowen's disease.The development of extragenital Bowen's disease may be associated with the infection with high-risk mucosal HPV types in some patients.

Objective To assess the relationship between human papillomavirus (HPV) and extrageni-tal Bowen's disease. Methods Regular PCR with consensus primers for LI region as well as mix primers and nested PCR were performed to detect the DNA of a broad range of cutaneous and mucosal HPV types in tissue samples from lesions of 41 patients with extragenital Bowen's disease and from normal skin of 48 human controls. Semiquantitative PCR and tyramide-based in situ hybridization (ISH) were also conducted to determine the load and localization of HPV DNA in HPV-positive samples. Results HPV DNA was detected in lesions from 5 (12%) of the 41 patients with extragenital Bowen's disease. Of the 5 HPV-positive patients, 3 carried mucosal HPV types (HPV16 in 2 cases, HPV 33 in 1 case) with a viral load of 101 to 103 copies, 2 cutaneous HPV types (HPV27 in 1 case and HPV76 in 1 case). As ISH showed, there was a generalized expression of mucosal HPV DNA in most tumor cell nuclei but not in peritumoral normal tissue, and no expression of cutaneous HPV DNA was observed in lesions. HPV DNA was detected in 1 (2.1%) control tissue sample, which proved to be epidermodysplasia verruciformis-associated HPV23. There was no significant difference in the detection rate of HPV DNA between the patients and controls. The viral load of cutaneous HPV types amounted to 10-2 to 10-3 copies in the 2 patients, which was similar to that of HPV 23 in the normal control. Conclusion Mucosal HPV types may be closely associated with the development of extragenital Bowen's disease.

Objective To survey the status of the clinical competence and meta-cognitive ability of college nursing students, to explore the correlation between the two factors, and analyze the influencing factors of baccalaureate nursing students′ clinical skills, then explore the approach of improving the clinical competence of nursing students. Methods Totally 184 nursing students from Taishan Medical University were investigated by General Information Questionnaire, Clinical Competence Test Scale, University Students Meta Cognitive Ability Scale. Results Nursing students′ average score of meta cognition was 80.48 ± 11.97, the average score of clinical competence was 121.22 ± 22.74. The meta cognitive ability and clinical competence of nursing undergraduate students both maintained at the middle level. Meta cognitive ability was positively correlated with clinical competence. To some extent, enhancing the meta cognitive ability could help students improve the clinical competence. Conclusions To understand the status and correlation between meta cognitive ability and clinical competence can guide students to learn effectively, can help teachers play a good role in teaching effectively.

Objective To investigate the scientific research ability and training needs of the new posts of medical undergraduates for better training them.Methods A total of 62 new posts of medical undergraduates from a level-two general hospital in China were recruited by questionnaire survey.Resuits the scientific research ability of the new posts of medical undergraduates is poor,so it is very necessary for their in-service training,93.5~ of them have interest in scientific research and hope to get the training,especially want to take part in the practical activity.Conclusions It is not better that the scientific research ability of the new posts of medical undergraduates,so as early as possible to give the scientific research skill training to them.

Objective To investigate the changes of IL-8 in peripheral blood of patients with pustular psoriasis after acitretin treatment.Methods Dual antibody sandwich enzyme-linked immunosorbent assay (ELISA)was performed to measure the IL-8 levels in the peripheral blood of 30 patients with pustular psoriasis and 30 patients with psoriasis vulgaris before and after treatment,and in 30 normal human controls.The relationship between the IL-8 level and disease severity was assessed for patients with pustular psoriasis.Results After acitretin treatment,the condition was improved greatly in patients with pustular psoriasis,together with a significant decrease in the level of IL-8 in the peripheral blood(57.07 ± 12.02 pg/ml vs.96.84 ± 14.68 pg/ml,P ＜ 0.05).Conclusion IL-8 may be involved in the therapeutic mechanism of acitretin in pustular psoriasis.

Objective To investigate the effects of low-dose of hydrocortisone on circulating thymus-dependent lymphocyte (T lymphocyte) apoptosis in patients with septic shock. Method fifty-seven patients with septic shock admitted into ICU from January 2006 to January 2009 were prospectively randomized (random number) to treatment group and control group. Another 20 healthy volunteers and 18 patients with sepsis alone were included as external control groups.The patients of treatment group and control group were treated with low-dose of hydrocortisone and placebo,respectively. Samples of peripheral blood were taken from healthy volunteers and patients 0 hr,24 hrs,48 hrs,72 hrs and 168 hrs after onset of the disease to determine the circulating T lymphocyte apoptosis by using the assays of Annexin V and flow cytometry. Least significant difference t -test was used for multiple comparisons. Results The percentage of Annexin V-positive CD4+ T lymphocytes in the primary stage was (11.01 +4.52)% in septic shock patients, (4.41 + 1.45)% in healthy volunteers, and (7.87 + 3. 82)% in patients with sepsis alone. And in the initial setting, the percentage of Annexin V-positive CD4+ T lymphocytes in the septic shock patients was higher than that in healthy volunteers ( P < 0.05) and in patients with sepsis alone ( P < 0.05). The percentage of Annexin V-positive CD8 + T lymphocytes at the beginning was (11.33+19.62)% in septic shock patients, (9.62+8.32)% in healthy volunteers, and (13.09+ 15.84)% in patients with sepsis alone (P > 0.05 between three groups). The percentages of Annexin V-positive CD4+ T lymphocytes in control group after 24 his, 48 hrs and 72 hrs were(13.51+6.85)%, (19.39 + 6.63)% and (15.33+ 6.21)%, respectively. And the percentages of Annexin V-positive CD4+ T lymphocytes in treatment patients after 24 hrs, 48 hrs and 72 hrs were (17.4 + 7.21)%, (22.61 + 5.64)%, and (25.73 + 6.91)%, respectively. The percentage of Annexin V-positive CD4+ T lymphocytes in septic shock patients was higher than that in control groups ( P < 0.05). The percentages of Annexin V-positive CD8+ T lymphocytes in control group after 24 hrs, 48 hrs and 72 hrs were (11.49+ 11.73)%, (12.74+ 10.39)% and (13.28+ 16.6)%, respectively, and in the treatment group, those were (9.49 + 8.9)%, (15.32+18.17)% and (13.68+16.84)%, respectively (P >0.05 between two groups). In the meantime, the percentages of Annexin V-positive CDS'1' T lymphocytes in control group and in treatment group were (12.72+ 19.69)% and (13.88 + 13.28)%, respectively (P >0.05). Conclusions Low-dose of hydrocortisone could induce CD4+ T lymphocyte apoptosis and has no effects on CD8+ T lymphocyte apoptosis when it is used to treat septic shock.

BACKGROUND:Study on antisense drug is still one of hotspots in the current field of biomedicine. Due to high-efficiency and specificity, antisense drug used for gene therapy has been paid more attention by many scholars.OBJECTIVE: To observe the effect of liposome-mediated keratin 14 (K14) antisense oligonucleotide on K14 gene and protein expressions as well as in vitro proliferation activities in human keratinocytes (KC).DESIGN: Single sample observation.SETTING: Department of Dermatology, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: Human KC, K14 oligonucleotide gene fragments (modified with phosphrothioate, and above sequence was synthesized by Shanghai Shenggong Bioengineering Company). Reverse transcriptase and TaqDNA polymerase were purchased from Invitrogen Company, K14 monoclonal antibody was purchased from Antibody Company, and SABC kit was purchased from Boster Company. EPICS-PRO-FILE Ⅱ flow cytometer was purchased from Coulter Company (USA).METHODS: Human epidermal KCs were primarily cultured, and their 3rd to 10th generations were used for the experiment.Artificially synthesized sense and antisense as well as mismatched K14 oligonucleotide gene fragments were introduced into KCs by means of liposome. Blank control group were set. The effects of antisense oligonucleotide on the cell cycle,K14 gene and protein expressions of KCs were detected by flow cytometer, reverse transcription polymerase chain reaction and SABC methods.MAIN OUTCOME MEASURES: Effect of oligonucleotide transfecting human KCs on the proliferation of KCs and K14 expression.RESULTS: [1]The electrophoresis of reverse transcription polymerase chain reaction products: Specific K14 gene band appeared in each group, and K14 gene expression in the antisense group was significantly lower in the sense group,missense group and blank control group. K14/β-actin value was similar among sense group, missense group and blank control group (P ＞ 0.05), But K14/β-actin value was significantly lower in the antisense group than in the above-mentioned 3 groups (F =47.554, P ＜ 0.01). ②K14 protein expression detected by immunohistochemical method:K14 was expressed in all the cultured KCs at different levels, and was obviously reduced after antisense oligonucleotide being added. 20 μmol/L antisense oligonucleotide could markedly inhibit K14 expression; K14 expression did not change in the control group. ③ DNA level change detected by flow cytometer: After being treated by K14 antisense oligonucleotide for 48 hours, human epidermal KCs were significantly increased at G1 stage (74.6%), and were markedly decreased at S stage (19.4%). Such changes were not found in the antisense group, missense group and blank control group.CONCLUSION: Antisense oligonucleotide can specifically inhibit K14 synthesis, and thereby, inhibit the proliferation of human KCs.

Objective To observe the changes of telomere length and MMPs level in human fibroblasts induced by UVB, and to explore their roles on skin photoaging .Methods Human skin fibroblasts were extracted and cul-tured.The 5th fibroblasts were irradiated by UVB .The morphology of fibroblasts were microscoped , and the length of telomere and the mRNA expression of COL1a1 and hTERT were detected by RT-qPCR.The expression of MMP-3 and MMP-1 were detected by Western blot .Results The fibroblasts gradually became round , wrinkled and disorderly arranged after 30 mJ/cm2 UVB irradiation for 24 h.The mRNA level of COL1a1 and hTERT and the expression of MMP-3 and MMP-1 were significantly increased after UVB irradiation compared with control , and the length of telomere was shortened .Conclusions UVB may frigger the early process of photoaging by the morphologi-cal changes of human skin fibroblasts and increasing the expression of MMP-3 and MMP-1 .

Objective To explore the significant risk factors in patients with Tile C pelvic frac-tures.Methods We conducted a retrospective review of all patients with Tile C pelvic fractures in Nanjing First Hospital from January 2010 to December 2014.The data gathered on each patient in-cluded:age,sex,mechanism of injury,visiting time after injury,shock index,Injury severity scale (ISS),Revised trauma score (RTS),Glasgowcoma scale (GCS),lowest PaO2/FiO2 ,6 h lactate clearance rate,concomitant injures and interventious of Tile C pelvic fractures.The data were tested using Student’t-test,χ2 test and logistic regression method.Results The study include 139 consecutive patients.The total mortality was 29.5%.Among them,36 (25.9%)patients died within 48 hours after admission.Multivariate regression analysis showed that shock index (OR=2.591,95%CI 1.041-4.216), ISS (OR = 47.96,95%,CI 15.89-147.23 ),RTS (OR = 6.917,95% CI 1.147-13.862 ),GCS (OR =4.172,95%CI 2.962-6.268),lowest PaO2/FiO2 (OR= 117.016,95% CI 51.011-176.032),6 h lactate clearance rate (OR=2.785,95%CI 1.191-4.892),concurrent head (OR=6.302,95%CI 2.270-13.175)or chest (OR=12.233,95%CI 5.193-33.985)injures were associated with high morality of Tile C pelvic frac-ture (P <0.01).The performing digital subtraction angiography with intravascular embolization can cut the mortality (OR=0.887,95%CI 0.875-0.899).Conclusion In our study,high trauma score,serious shock, coma,PaO2/FiO2 decreased and 6 h lactate clearance rate decreased,combined with the head and chest inju-ry are the important reasons of mortality in patients with Tile C pelvic fracture.It’s vital to control shock actively,use trauma scale and emphasize multidisciplinary cooperation to reduce mortality in patients with Tile C pelvic fractures.

Objective To compare the different characteristics of adaptive support ventilation (ASV) and synchronized intermittent mandatory ventilation-pressure support ventilation (SIMV-PSV) mode in weaning patients after general anesthesia.Methods One hundred and twenty-eight patients received general anesthesia,ending in odd and even numbers by hospital number divided into ASV group (single number,62 cases) and SIMV-PSV group (double number,66 cases).The propofol dosage,duration of mechanical ventilation,duration of intubation,ventilator alarms,ventilator settings manipulations and each stage of the blood gas analysis,hemodynamic,respiratory mechanics were recorded.Results One hundred and twenty-eight patients completed the extubation.The propofol dosage,duration of mechanical ventilation,duration of intubation in ASV group were significantly shorter than those in SIMV-PSV group [(1.13 ± 0.33)mg/kg vs.(1.28 ±0.49) mg/kg,(169.8±36.5) min vs.(201.9 ±37.3) min,(197.2 ±38.9) min vs.(239.5 ± 42.3) min,P ＜ 0.05].There was no statistically significant difference in various stages of heart rate,mean arterial pressure,central venous pressure,pH,arterial carbon dioxide partial pressure,oxygenation index between two groups (P ＞0.05).In the first and second stages,tidal volume in ASV group was significantly higher than that in SIMV-PSV group [(543.6 ± 72.3) ml vs.(489.5 ± 68.7) ml,(513.9 ± 65.7)ml vs.(462.8 ± 61.7) ml,P＜ 0.05],respiratory rate in ASV group was significantly lower than that in SIMV-PSV group [(13.2 ± 3.6) times/min vs.(17.2 ±4.1) times/min,(15.1 ± 3.1) times/min vs.(16.8 ± 3.7)times/min,P ＜ 0.05].In the first stage,the mean airway pressure and peak airway pressure in ASV group were significantly lower than those in SIMV-PSV group [(8.2 ± 1.7) cm H2O (1 cm H2O =0.098 kPa) vs.(12.3 ± 3.1) cm H2O,(16.2 ± 2.9) cm H2O vs.(21.2 ± 4.3) cm H2O,P ＜ 0.05].The pulmonary dynamic compliance in ASV group was better than that in SIMV-PSV group [(64.8 ± 12.3) ml/cm H2O vs.(52.6 ±13.5) ml/cm H2O,P ＜ 0.05].The ventilator alarms,ventilator settings manipulations in ASV group were significantly shorter than those in SIMV-PSV group [(2.3 ± 1.6) times vs.(5.1 ± 1.9) times,(0.8 ± 0.5) times vs.(1.6 ± 0.8) times,P ＜ 0.05].Conclusion ASV weaning mode is safe and effective,which could accelerate the extubation and simplify the manipulation.