Objective:To investigate the immuno-effects triggered by dendritic cells (DCs) pulsed with Heat-treated lymphoma cells.Methods:Mononuclear cells from healthy human peripheral blood(PBMC) were isolated and DCs were generated in vitro with normal methods.Lymphoma cells were heated (42℃,2 h) and then loaded onto DCs.Immunotypes of DCs and lymphocytes were detected with flow-cytometric analysis.IFN-γ releasing was detected by ELISPOT,lymphocyte cytotoxicity was analyzed by MTT method after mixing with DC reaction.Results:After pulsing DCs with hyperthermia lymphoma cells,CD80~+,CD86~+ and HLA-DR existed higher than those of the control group,IFN-γ releasing and cytotoxicities were also strengthened,but there was no difference between them,cytotoxitic lymphocytes (CTLs) were produced more as CD8~+,CD25~+ and CD56~+ cells by flow-cytometric analysis.Conclusion:Anti-lymphoma immuno-effects could be strengthened by loading DC with hyperthermia lymphoma cells.

Objective To investigate the DNA methylation feature and DNA methylation regulation to its transcription and expression of O6-methylguanine-DNA methyltransferase gene (MGMT) in NaAsO2-treated HaCaT cells. Methods HaCaT cells were treated 72 hours at intervals and repeatedly by 3.13, 6.25,12.50, and 25.00 μmol/L NaAsO2, MGMT gene promoter region was amplified in the transcription initiation site - 329 - + 93 region by bisulfate-sequencing polymerase chain reaction (BSP), the mRNA transcription and the protein expression of MGMT was detected by real-time quantitative PCR and Western blotting. NaAsO2-untreated HaCaT cell was set as a blank control, and human epidermal squamous carcinoma cell strain A431 was set as a positive control. Results Among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, the positive rates of the DNA methylation of promoter region in MGMT gene were 0.63%(l/160), 6.25% (10/160), 10.63%( 17/160) and 18.75% (30/160), respectively, and methylated CpG sites were mainly located in - 249--146 region relative to transcription start site. There was no DNA methylation in the blank control. There were significant differences between the blank control and the NaAsO2-treated cells (x2 = 76.687, P< 0.05). Average levels of MGMT mRNA were 1.518 31 ± 0.180 54, 1.425 22 ± 0.180 39, 1.014 54 ± 0.096 79 and 0.887 72 ± 0.020 00, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells(1.198 29 ± 0.159 97), there were significant differences(F = 37.359, P < 0.05). Average levels of MGMT protein were 1.174 47 ± 0.064 75, 0.848 83 ± 0.057 01, 0.471 63 ± 0.023 34 and 0.240 34 ± 0.014 43, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells (1.066 19 ± 0.061 24), there were significant differences(F = 20.687, P < 0.05). Conclusions Arsenic can cause CpC island hypermethylation in the promoter region of MGMT gene, which results in inhibited MGMT mRNA transcription and protein expression. It might be one of the important mechanisms of arsenic-induced skin lesion.

objective: To improve the repair technique of secondary nasal deformity following surgical treatment in the patients with bilateral cleft lip. Methods: 12 cases were treated with the operation technique: bilateral vermilion flaps were inserted into prolabial for the enlongation of upper lip and "V Y incision suture" principle was employed to repaire the nasal deformity. Results: Satisfactory results were achieved in all the 12 cases. Conclusion: The technique is feasible in the reconstruction of Cupid's bow, philtrum and correction of nasal and labia deformities.

OBJECTIVE:To observe therapeutic efficacy and safety of Kouyanqing granules for chronic periodontitis and it effects on inflammatory cytokine. METHODS:110 patients with chronic periodontitis were divided into control group(55 cases)and observa-tion group(55 cases). Control group accepted the periodontal basic treatment as oral hygiene instruction,supragingival scaling,subgin-gival scaling and root planning. Observation group was additionally given Kouyanqing granules 20 g orally,twice a day,on the basis of control group. Both groups were treated for 4 weeks. Clinical efficacies of 2 groups were observed as well as the levels of plaque in-dex(PLI),gingival index(GI),probing depth(PD),clinical attachment level(CAL),IL-17,IL-21,and the occurrence of ADR. RE-SULTS:Both groups completed the treatment,and no patient lost to follow up. Total response rate of observation group was significant-ly higher than that of control group,with statistical significance(P<0.05). After treatment,the levels of PLI,GI,PD,CAL,IL-17 and IL-21 in 2 groups were significantly lower than before,and the observation group was significantly lower than the control group, with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Based on routine treatment,Kouyanqing granules show significant therapeutic efficacy for chronic periodontitis, can significantly improve periodontal symptom and reduce inflammatory factor level,but do not increase the incidence of ADR.

Objective To investigate the transcription and expression of DNA methyltransferase 1 (DNMT1) mRNA in endemic arsenism patients by burning coal usage,to probe its effects on the development and carcinogenesis of arsenism. Methods In 2008,68 arsenism patients(including 24 mild cases,28 moderate cases and 16 severe cases) were selected in the areas with endemic arsenism according to Standarding of Diagnosis for Endemic Arsenism from Xingren county,Guizhou province. Among the subjects,40 cases were diagnosed by pathological methods,and they were divided into general pathological changes(20),precancerous(14) and cancerous group(6). Tweleve kilometer away from the endemic arsenism area,23 controls were selected in Daguoduo village (non-arsenism exposure). Under the principle of informed consent,blood samples were collected from individuals. The mRNA expression of DNMTI was detected by real-time quantitative reverse transcription polymerase chain reaction(FQ-PCR). At the same time,skin tissue samples were collected from the voluntary surgical treatment patients with endemic arsenism (total 61 cases,including 34 general pathological changes cases,21 precancerous cases and 6 cancerous cases) and from the control(15 cases). DNMT1 protein was detected by immunohistochemical method.Results Average level of DNMT1 mRNA were 0.221 83±0.595 09,0.246 11±0.509 79 and 0.389 27±0.411 33 respectively among mild,moderate and severe arsenism group. DNMT1 mRNA level of mild and moderate group were obviously lower than the control group(0.695 95±0.463 98,all P < 0.01). The mRNA average level of DNMT1 were 0.320 64±0.547 46,0.313 09±0.529 13 and 0.159 07±0.342 56 individually among general pathological changes,precancerous and cancerous group,which were obviously lower than the control group(0.695 95±0.463 98,all P < 0.05). The expression rates of DNMT1 protein in skin were 88.24%(30/34),100%(21/21) and 100% (6/6) among general pathological changes,precancerous and cancerous group were higher than the control group [0(0/15),all P < 0.01],and the extent of expression gradually increased with the aggravation of skin damage(r,= 0.740,P < 0.01). Conclusions DNMT1 participated in the development of the arsenism. High expression of its protein was an early event during the process of the arsenism. DNMT1 may be the new target markers for early diagnosis and treatment of arsenism.

Objective To investigate and analyse the current situation and the factors impacting nursing soft skills in order to provide targeted advice to improve students′ nursing soft skills .Methods Totally 1 000 student nurses were investigated by the nursing soft skills scale and the self‐designed questionnaire .Results First ,there was no differences between the nursing soft skill of students and the national new nurses(P> 0 .05) .But the students′ organizational management and cognitive skills were lower than that of national new nurses(P< 0 .05) .Second ,class leader ,familiarity and fancy for nursing science were the main factors impac‐ting nursing soft skills(P< 0 .05) .Third ,the students′ organizational management and cognitive skills had an increasing trend fol‐lowing the extent of familiarity and fancy for nursing science(P< 0 .05) .Conclusion Education management institution and nursing schools should try to make some reform mearsure to improve students′ nursing soft skills .

AbstractBACKGROUND: Fas and P53 are important regulator and control gene which can promote apoptosis. They belong to the receptor family part of tumor necrotic factor/nerve growth factor. Their expression products have effects on apoptosis signal transmission, and can regulate and control cell apoptosis in cerebral ischemia-reperfusion injury. And puerarin can alleviate the level of cell apoptosis.OBJECTIVE: To observe the effect of puerarin on Fas and P53, the apoptosis-related gene of nerve cell in hippocampns CA1 region of rats af ter cerebral resuscitation.DESIGN: Randomized controlled trial. SETTING: Department of Emergency, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: The experiment was carried out at Emergency Department, Tongji Hospital, Tongji Medical College, Huazhong University ofScience and Technology from September 2001 to Februray 2002. Totally 45 of 3 months old Wistar rats of clean grade were selected, and randomly divided into 3 groups: sham operation group, model control group and puerarin treatment group with 15 rats in each group.METHODS: Acute global brain ischemia-reperfusion models were established in rats of puerarin treatment group and model control group. In rats of sham operation group, stigmata of both flanks of the first cervical vertebrae were isolated, but bilateral vertebral arteries were not electric coagulated, and biolateral common carotid arteries were only isolatedwithout clamping close. Rats in puerarin treatment group were given puerarin injection 100 mg/kg, 1 hour before ischemia, and model control group were given normal saline in equivalence while rats in sham operation group were not given medicine. Death of rats in each group was performed separately in the 3rd, 6th, 12th, 24th and 48th hours after cerebral ischemia-reperfusion with 3 rats per group in each time. Hippocampus tissues of rats were isolated, and tissue slices were preparated. And the changes of- the protein expression levels and the number of apoptosis cells of rats in each group at different time point after cerebral ischemia-reperfusion were detected in immuno-histochemical method and end labelling in situ method. MAIN OUTCOME MEASURES: ① The number of positive cells inprotein expression of Fas and P53 in hippocampus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion was studied. ② Comparison of the number of apoptosis cells in hippocampus CA1 region of rats between groups at different time point after cerebral ischemia-reperfusion were studied, too.RESULTS: All the 45 rats enrolled in research were entered the stage of result analysis: ① The number of positive cells in protein expression of Fas in hippocampus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion: Obvious gene expression of Fas was not found in sham operation group. In contrast with model control group, obvious decrease was found at all time points after cerebral ischemi a-reperfusion in puerarin treatment group, and in the 6th, 12th, 24th and 48th hour the differences were significant [(15.0±4.3), (13.5±4.9); (40.7±3.4), (27.2±3.1); (37.0±4.8), (22.0±2.1); (24.7±4.1), (18.9±5.3)/mm; P < 0.05,P < 0.01]. ② The number of positive cells in protein expression of P53 in hippocanpus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion: Obvious gene expression of P53 was not found in sham operation group. In contrast with model control group, obvious decrease was found in the 24th and 48th hour after cerebral ischemiareperfusion in puerarin treatment group [(25.3±4.4), (12.8±2.7); (24.3±3.6), (10.9±3.0)/mm; P < 0.01]. ③ Comparison of the number of apoptosis cells in hippocampus CA1 region of rats between groups at different time point after cerebral ischemia-reperfusion :In contrast with model control group,obvious decrease was found in the 12th, 24th and 48th hour after cerebral is chemia-reperfusion in puerarin treatment group [(34.0±3.7), (21.0±3.7); (41.0±4.2), (33.0±4.8); (71.0±5.5), (41.0±3.4)/mm; P < 0.01].CONCLUSION: In rats which were given puerarin treatment, the expression of Fas decrease obviously in 6 to 48 hours after cerebral ischemiareperfusion, and the expression of P53 decreased obviously in the 24th to 48th hour after cerebral ischemia-reperfusion, and a descent tendency could be found in the number of apoptosis cells. These can further prove the cerebral protective effect of puerarin, and indicate that the inhibition of puerarin to cell apotosis after cerebral resuscitation is related to its effect on the decrease in protein expression of apoptosis-promoting gene, Fas and P53.Puerarin has a protective effect on cerebral ischemia-reperfusion injury of rats. In comparison with model control group, the expression of Fas in puerarin treatment group has an obvious decrease inthe 6th to 48th hour after cerebral ischemia-reperfusion, the expression of P53 has an obvious decrease in the 24th to 48th hour after cerebral ischemia-reperfusion, and the number of apoptosis cells decrease obviously, too, which further improves the cerebral protective effect of puerarin and indicates that the inhibition of puerarin to cell apoptosis after cerebral resuscitation is related to its effect on the decrease in protein expression of apoptosis-promoting gene Fas and P53.

Objective To investigate the relationship between ultrasonographic appearance of the endometrium and steroid. Methods A total of 30 women with normal cycle were used for this study. On the day of positive urinary LH, the subjects were classified into group A, group B and group C according to Gonen's criterions of endometrial echogenic patterns. Blood samples were taken for measurement of FSH, LH, PRL, E_2, P, T levels on the third day of the cycle, the pre-ovulatory phase, and the sixth or seventh day after ovulation. Results During the early follicular phase and pre-ovulatory phase, serum E_ 2 concentration was highest in group A, then group B and lowest in group C; there was significant difference between group A and group C (P

0.05);the disappearance rate of varices of fundus of stomach and esophagus in the combined devascularization group was higher than that in control group(P0.05).The long-term(fourth to seventh year)survived rate of the combined devascularization group was higher than that of control group(98.3% in the combined devascularization group,90.0% in control group)(P

Objective To investigate the effect of NaAsO2 on the binding levels of methyl CpG binding protein 2(MeCP2),DNA methyltransferase 1 (DNMT1) and histone deacetylase 1(HDAC1) to the hypermethylation promoter region of MGMT gene in HaCaT cells,in order to provide a basis to deepen the interpretation of the role of arsenic poisoning mechanism.Methods HaCaT cells were treated repeatedly and interval with different concentrations of NaAsO2(3.13,6.25,12.50,25.00 μnol/L,respectively) for 72 h.Untreated HaCaT was used as blank control group and human epidermal squamous carcinoma cell line(A431 cells) as positive control group.The binding levels to the two transcription regulatory regions(ChIP1,ChIP2) and to the coding region(ChIP3) of MGMT 8ene were detected by chromatin immuno-precipitation combined with quantitative PCR.Results The differences of binding levels of MeCP2,DNMT1 and HDAC1 to ChIP1 and ChIP2 in each group were significant (F=7.387,84.634,78.442 and 19.263,69.649,26.546,all P ＜ 0.05).The binding levels of MeCP2,DNMT1 and HDAC1 to ChIP1 and ChIP2 in each NaAsO2 exposed group[3.13 μmol/L NaAsO2 exposed group:(136.00 ±16.97)％,(145.00 ± 2.83)％,(88.50 ± 19.09)％ and (106.50 ± 37.48)％,(112.34 ± 8.73)％,(59.71 ± 8.49)％;6.25 μmol/L NaAsO2 exposed group:(130.00 ± 42.43)％,(154.50 ± 4.95)％,(101.00 ± 1.27)％ and (88.50 ±3.54)％,(134.32 ± 2.82)％,(102.75 ± 19.91)％ ; 12.50 μmol/L NaAsO2 exposed group:(141.50 ± 23.33)％,(161.50 ± 7.78)％,(125.00 ± 11.31)％ and (119.50 ± 24.75)％,(171.59 ± 3.54)％,(167.61 ± 10.61)％; 25.00μmol/L NaAsO2 exposed group:(134.50 ± 43.13)％,(472.50+ 50.20)％,(383.50 ± 30.41)％ and (180.09 ±12.73)％,(348.50 ± 27.58)％,(158.45 ± 12.02)％] were higher than that in the blank control group[(51.50 ±9.19)％,(82.00 ± 12.73)％,(25.03 ± 2.91)％ and (37.02 ± 4.24)％,(91.56 ± 26.16)％,(19.09 ± 2.90)％,all P ＜ 0.05].The differences of binding levels of MeCP2 to ChIP3 in each group were not significant(F =1.670,P ＞0.05),but the differences of binding levels of DNMT1 and HDAC1 to ChIP3 were significant (F =4.404,9.863,all P ＜ 0.05),and only the binding levels in the 25.00 μmol/L NaAsO2 exposed group [(615.85 ± 29.63)％,(306.09 ± 59.40)％] were higher than that in the blank control group[(99.70 ± 12.02)％,(92.45 ± 48.79)％,all P ＜ 0.05].Conclusions MeCP2 can bind to the methylated MGMT gene transcriptional regulatory regions which are induced by arsenic and leads to histone deacetylation by the recruitment of DNMT1 and HDAC1 and,meanwhile,DNMT1 can bind to the coding region of MGMT gene to recruit HDAC1 in a methyl DNA binding protein(MBD) independence manner and media MGMT gene silencing through the chromatin remodeling way,which might be the early molecular events of arsenic poisoning.