The 5-methylcytosine (~mC) in DNAs from 5-azacytidine and MNNG treated FL, Wish and Veto-E6 ceUs were analysed by HPLC. In 2?10~(-6)mol/L 5-azaCR treated cells, the percentages of ~mC in total cytosine were all lowered significantly (P 0.05). These results were in good agreement with those obtained by radioactivity analysis of newly replicated DNA fragments from Hpa Ⅱ digest. These results further validate the idea that DNA hypomethylation as a general pathway in the initiation process of chemical carcinogenesis is based on the results obtained by a defectively designed experiment.

Background Researches determined that ELOVL4 gene is a disease-causing gene of Stargard tmacular dystrophy and is a elongation enzyme of very long chain fatty acids.Stargardt type Ⅲ(STGD3) may be associated with the metabolism of extensive enzyme of very long chain fatty acids.To explore the effect of ELOVL4 in STGD3 and its relationship with the metabolism of extensive enzyme of very long chain fatty acids is of important clinical significance.Objective This study was to determine the role of ELOVL4 gene for the pathogenesis of STGD3.Methods Recombinant adenovirus type 5 carrying mouse ELOVL4 gene and green fluorescent protein (GFP) was transfected into pheochromocytoma cells (PC12 cells),and then the cells were divided into PC12 group,PC12+Ad5-GFP group and PC12+AdS-ELOVL4 group.Ad-GFP or Ad-ELOVL4 was added into the culture medium for 24 hours with the virus plasmid 1 × 104-2× 104 pfu/ml.Expression of ELOVL4 mRNA in the PC12 was quantified by quantitative real time PCR(qRT-PCR) and was described as relative value to the expression of RPL19.ELOVL4 protein was assayed by Western blot.The transfected cells were treated with arachidonic acid (AA,20:4n6),eicosapentaenoic acid (EPA,20:5n3) and docosahexaenoic acid (DHA,22:6n3) individually for 48 hours.The cells were collected,and total lipids were extracted,and fatty acid methyl esters were prepared and analyzed by gas chromatography-mass spectrometry (GC-MS).Results The relative expression levels of ELOVL4 mRNA in PC12 cells in the PC12+Ad5-ELOVL4 group,PC12+Ad5-GFP group and PC12 group were 0.833± O.138,0.027t±0.002 and 0.024 ±0.002,with a significant difference among the 3 groups (F =102.700,P =0.000),and relative expression levels of PC12+Ad-ELOVL4 were 30-40 folds more than those in the PC12 group and the PC12+Ad-GFP group.Western blot assay showed a stronger response band for ELOVL4 protein in the PC12+Ad-ELOVL4 group.GC-MS found that abundant polyunsaturated fatty acids (C28-C38) were synthesized by PC12 cells in the PC12+Ad-ELOVL4 group,with the more levels in C34 and C36.Conclusions ELOVL4 can promote the synthesis of C28-C38 polyunsatured fatty acid in PC12 cells,which offers a novel clue for the treatment of STGD3.

Objective:To explore the related factors of postoperative adjuvant therapy for cervical cancer stagedⅠB1-ⅡA2 [according to 2018 International Federation of Gynecology and Obstetrics (FIGO) staging standard], and to establish a nomogram model to predict the risk of postoperative adjuvant therapy for locally advanced cervical cancer.Methods:A total of 714 patients with cervical squamous cell cancer staged FIGO ⅠB1-ⅡA2 treated by surgery in Anhui Provincial Hospital were selected as the research objects from January 2009 to December 2019, and their clinicopathological data were analyzed. Multiple logistic regression analysis was used to determine the influencing factors, and a nomogram model was established to predict the risk of postoperative adjuvant treatment of cervical cancer. The predictive performance of the model was evaluated with the consistency index (C-index), and the compliance of the model was evaluated with the calibration curve.Results:Univariate analysis suggested that postoperative adjuvant therapy for cervical cancer was associated with gravidity ( χ2=11.506, P=0.001), underlying disease (hypertension or diabetes) ( χ2=7.668, P=0.006), squamous cell cancer antigen (SCC-AG) level ( χ2=19.392, P<0.001), imaging risk factors ( χ2=16.392, P<0.001), FIGO stage ( χ2=25.686, P<0.001), tumor size ( χ2=9.392, P=0.025) and surgical path ( χ2=16.590, P<0.001). Multivariate logistic regression analysis suggested that the number of pregnancy >2 times ( OR=1.951, 95% CI: 1.355-2.808, P<0.001), SCC-Ag ≥1.5 μg/L ( OR=2.021, 95% CI: 1.444-2.829, P<0.001), FIGO stage ⅠB3-ⅡA2 [ⅠB3 ( OR=1.933, 95% CI: 1.139-3.282, P=0.015); ⅡA1 ( OR=2.723, 95% CI: 1.556-4.765, P<0.001); ⅡA2 ( OR=3.159, 95% CI: 1.502-6.646, P=0.002)], with underlying disease (hypertension or diabetes) ( OR=1.867, 95% CI: 1.051-3.318, P=0.033), imaging risk factors ( OR=1.997, 95% CI: 1.127-3.537, P=0.018), without neoadjuvant therapy [preoperative neoadjuvant therapy for 1 cycle ( OR=0.402, 95% CI: 0.207-0.783, P=0.007)] and laparoscopic surgery ( OR=2.177, 95% CI: 1.524-3.112, P<0.001) were independent influencing factors for postoperative adjuvant treatment of cervical cancer. Based on the screened variables, the nomogram model to predict the risk of postoperative adjuvant treatment for cervical cancer has good predictive performance (C-index was 0.702) and compliance. Conclusion:The number of pregnancy >2 times, SCC-Ag ≥1.5 μg/L, FIGO stage ⅠB3-ⅡA2, with underlying disease (hypertension or diabetes), imaging risk factors, without neoadjuvant therapy, and laparoscopic surgery are independent influencing factors for postoperative adjuvant treatment of cervical cancer. A nomogram model has been constructed to predict the risk of postoperative adjuvant therapy for locally advanced cerrical cancer, and it can provide evidence for clinical treatment selection.

Objective To investigate the estimated values of sequential organ failure assessment (SOFA) and quick SOFA (qSOFA) for diagnosis and prognosis in patients with sepsis according to the new diagnostic criteria in Sepsis 3.0.Methods A retrospective study was conducted.All the clinical data were collected from patients with definite diagnosis of infection and they were admitted into the Intensive Care Unit (ICU) of Beijing Traditional Chinese Medicine Hospital Affiliated to Capital Medical University from July 2014 to June 2016.The patients' gender,age,infectious location,respiratory rate (RR),oxygenation index (PaO2/FiO2),Glasgow coma scale (GCS),total bilirubin (TBil),platelet count (PLT),serum creatinine (SCr),serum lactate level,etc.general data on admission were collected to carry out SOFA and qSOFA scorings.And then the septic patients in accord with the diagnostic criteria of Sepsis 3.0 were screened out.According to outcome after admission,the septic patients were divided into survival group and death group,and the differences in diagnosis and in estimation value of prognosis between SOFA scoring and qSOFA scoring were assessed as SOFA group and qSOFA group.Results From 545 septic patients enrolled,189 septic patients consistent with the diagnostic criteria of Sepsis 3.0 were selected.In SOFA scoring group,the morbidity of septic patients was 34.68％,while in qSOFA scoring group,it was 15.96％,the difference between the two groups being statistically significant (P ＜0.01).The mortality was significantly lower in SOFA scoring group than that in qSOFA scoring group [28.04％ (53/189)vs.42.53％ (38/87),P ＜ 0.05].The mortality of qSOFA scoring group was about 1.52 times that of SOFA scoring group.On the aspect of scoring,in patients with SOFA scoring the score of death group was significantly higher than that in survival group (8.74 ± 0.417 vs.7.10 ± 0.235,P ＜ 0.01);in the patients with qSOFA scoring,the score in death group compared with that in survival group showed uo statistical significant difference (2.32 ± 0.48 vs.2.16 ± 0.37,P ＞ 0.05).On the aspect of laboratory indexes,the levels of GCS score in death group was significantly lower than that in the survival group (8.15 ± 0.67 vs.12.48 ± 0.36),blood lactate level in death group was significantly higher than that in the survival group (mmol/L:8.55 ± 4.66 vs.2.31 ± 0.16,P ＜ 0.01);the PaO2/FiO2,TBil,PLT and SCr showed no significant differences between the two groups (all P ＞ 0.05).Conclusions The new diagnostic criteria (Sepsis 3.0) can be used for diagnosis of sepsis in ICU.Compared with qSOFA scoring,the SOFA scoring is more suitable to be used for diagnosis and predicting prognosis of septic patients in ICU;SOFA scoring,GCS scoring and serum lactate level can be applied to estimate outcome of septic patients.

OBJECTIVE To share surgical experience of ossicular chain malformations with CO2 laser.METHODS A retrospective analysis was performed.9 patients with clinical diagnosis of ossicular malformations underwent ossicular reconstruction with CO2 laser-assisted from May 2010 to Mar 2016,the results were evaluated by comparing preoperative and postoperative audiometric outcomes and the rate of postoperative complications.RESULTS 8 cases with complex lesion combining incus and stapes were found intraoperatively,the deformity located on stapes was showed in one case.The mean postoperative air conduction (AC) value was (26.53 ± 12.28) dB,the mean postoperative air-bone gap (ABG) was (9.44 ± 9.62) dB,the postoperative AC and ABG value improved considerably comparing with the preoperative value in all the patients,the difference was statistically significant.CONCLUTION CO2 laser is a suitable and effective adjunct in surgery for ossicular malformations such as otosclerosis.The use of the laser improves hearing results and operation efficiency and is not likely to increase side-effects to patients.

Objective To evaluate the efficacy and safety of lenalidomide in a real-world clinical practice in Chinese patients with multiple myeloma (MM).Methods It was a prospective,multi-center,observational study.A total of 165 consecutive patients with MM treated with lenalidomide-based regimens were enrolled in 12 hospitals from June 2013 to November 2015.Relevant information was recorded,such as baseline clinical data,cytogenetic abnormalities,treatment regimens,and duration of treatment,safety,and survival.Results (1)There were 126 relapsed and refractory MM (RRMM) patients,25 newly diagnosed patients and 19 maintenance patients.The evaluable RRMM patients accounted for 120 cases,among which 74 cases(61.7％) reached the partial response (PR) or above,and a very good partial response (VGPR) in 16 patients (13.3％),a complete response (CR) in 14 cases (11.7％),a strictly complete response (sCR) in 4 cases (3.3％).Thus,a VGPR or above in 34 patients accounted for 28.3％.(2)The median follow-up was 13 months,the median time to progression 12 months.The median survival after receiving lenalidomide was 19 months,and the median overall survival (OS) was 62 months.(3) The univariate analysis in 120 RRMM patients suggested that prognostic factors for significant improvement in PFS included normal karyotype,international staging system (ISS) Ⅰ-Ⅱ,t(4;14) negative (detected by fluorescence in situ hybridization),non-bortezomib resistance and response to previous regimens.As to OS,nonbortezomib resistance,response to previous regimens and non-primary refractoriness were positive factors.Multivariate analysis showed that the response to previous regimens (PR or better) was an independent good prognostic factor for progress-free survival (PFS),non-bortezomib resistance and non-primary refractoriness for OS.(4) Grade 3 or 4 adverse events that occurred in more than 10％ of all enrolled patients were neutropenia (12.7％),leukocytosis (11.5％) and thrombocytopenia (12.7％).Owing to intolerance of toxic side effects,7 cases withdrew lenalidomide.Conclusions No matter what combination,regimens containing lenalidomide are effective to RRMM patients with overall response rate 61.7％,a time to progression 12 months and an overall survival 62 months.The toxicity is quite tolerable and manageable.In addition,the response to previous treatment (reached PR or above) is the independent good prognostic factor for PFS,non-bortezomib resistance and non-primary refractoriness for OS.Clinical trail registration Clinicaltrials.gov,NCT01947309

Objective:To study the safety and efficacy of liquid isolation assisted technology in ultrasound-guided percutaneous radiofrequency ablation for hepatocellular carcinoma adjacent to liver surfaces.Methods:A retrospective study was conducted on 246 patients with hepatocellular carcinoma near liver surfaces treated by radiofrequency ablation from November 2016 to November 2019 at the Department of Hepatobiliary and Pancreatic Surgery, Affiliated Tumor Hospital of Zhengzhou University. There were 182 males and 64 females, aged (56.7±9.2) years. The patients were divided into the liquid isolation group with liquid isolation assisted technology for radiofrequency ablation ( n=131) and the conventional treatment group with conventional radiofrequency ablation ( n=115). The complication, one-time complete ablation and progression-free survival rates were compared between the two groups. Results:The incidence of intraoperative complications in the liquid isolation group was 21.4% (28/131), versus 20.9% (24/115) in the conventional treatment group. There was no significant difference between the two groups ( P>0.05). The incidence of postoperative complications in the liquid isolation group was 20.6% (27/131), which was significantly lower than that in the conventional treatment group 33.9% (39/115) ( P<0.05). The incidence of postoperative pain in the liquid isolation group was 6.1% (8/131), which was significantly lower than that in the conventional treatment group 15.7% (18/115) ( P<0.05). For the 131 patients in the liquid isolation group, there were 183 tumors near to liver surfaces. For the 115 patients in the conventional treatment group, there were 142 lesions near to liver surfaces. The one-time complete ablation rate in the liquid isolation group was 94.0% (172/183), which was significantly higher than the 75.4% (107/142) in the conventional treatment group ( P<0.05). The progression-free survival rate in the liquid isolation group was significantly better than that in the conventional treatment group ( P<0.05). Conclusion:The liquid isolation technique increased the one-time complete ablation rate of liver cancer adjacent to liver surfaces, reduced the incidence of postoperative complications and improved prognosis.

Objective:To evaluate the effect of canagliflozin on intrarenal fat content and oxygenation in newly-diagnosed type 2 diabetes patients.Methods:Twenty-three newly-diagnosed type 2 diabetes patients were divided into canagliflozin( n=11) and glimepiride control( n=12) groups .Both groups received MRI scanning with Dixon MRI and BOLD MRI sequence to assess patients′ intrarenal fat content, oxygenation level before treatment and 24 weeks after treatment. Fasting blood glucose, glycosylated hemoglobin, blood uric acid, blood lipids, blood pressure, weight, and other metabolic index were also tested before and after treatment. Furthermore, the relationship between body mass index(BMI) and intrarenal fat content and the correlation between changes in intrarenal fat content and improvement in renal hypoxia were analyzed. Results:No significant differences were found in baseline age, body weight, fasting blood glucose, glycosylated hemoglobin, blood lipid, and serum uric acid between the two groups. There was no significant difference in fasting blood glucose, glycosylated hemoglobin, cholesterol(CHO), low-density lipoprotein-cholesterol(LDL-C), and triglycerides(TG) levels in both groups after 12 and 24 weeks of treatment. The decrease in body weight, blood uric acid level, and diastolic blood pressure from baseline in the canagliflozin group was greater than those in the control group( P<0.05). Two groups of patients with type 2 diabetes at baseline had no obvious difference in intrarenal fat content, and the patients′ BMI showed no obvious correlation with degree of intrarenal fat accumulation. Canagliflozin treatment for 24 weeks could reduce intrarenal fat content, which was higher than that of control group. The R2 * values of renal cortex and medulla in the canagliflozin group decreased from baseline by 19.22% and 22.63% respectively( P<0.05), whereas no significant difference was seen in the glimepiride control group. The decrease of intrarenal fat content in the canagliflozin group was related to the improvement of renal cortex and medulla oxygenation. Conclusion:Canagliflozin can reduce intrarenal fat accumulation and improve renal cortical hypoxia in newly diagnosed type 2 diabetes patients with normal renal function.

ObjectiveTo observe the effect of transcranial direct current stimulation (tDCS) on motor function of upper limbs of stroke patients with hemiplegia. MethodsFrom October, 2020 to October, 2021, 65 patients from Wuhan No.1 Hospital were randomly divided into control group (n = 32) and observation group (n = 33). All the patients received routine rehabilitation and mirror therapy, and the observation group received tDCS in addition, for four weeks. They were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE), Action Research Arm Test (ARAT) and modified Barthel index (MBI) before and after treatment. ResultsThe scores of FMA-UE, ARAT and MBI improved in the both groups after treatment (|t| > 10.455, Z = -2.793, P < 0.001), and all the scores were better in the observation group than in the control group (|t| > 4.152, Z = -2.045, P < 0.05). ConclusionThe combination of tDCS can effectively promote the recovery of upper limb motor function of stroke patients.