Hyperhomocysteinemia, which is an independent risk factor for atherosclerosis, may cause aberrant methylation and dysregulation of gene expression, but the characteristics of the aberrant methylation and its key links involved in its pathogenic mechanisms are still poorly understood. The effect of hyperhomocysteine on DNA methylation in vascular smooth muscle cells, its characteristics and the underlying mechanism of Hcy-induced changing in DNA methylation patterns were investigated. Clinical relevant concentrations of homocysteine was added into the cultured vascular smooth muscle cells of the Homo sapien umbilical vein for 24 h. The level of SAM and SAH was detected by HPLC. The activity of SAH Hydrolase was detected by real-time quantitative reverse transcription-PCR and Western blotting analysis. The level and patterns of DNA methylation were measured by endogenous C-5 DNA methyltransferase(C-5 MT-ase) activity and capacity of genomic DNA to accept methyl groups and methylation-dependent restriction analysis. The results indicated that an increased Hcy concentration induced elevated SAH, declined SAM and the ratio of SAM/SAH, reduced expression of SAH Hydrolase, but increased activity of C-5MT-ase. The methylation status of gDNA analyzed by methyl-accepting capacity of gDNA uncovered a demethylation process in gDNA, or homocysteine-caused hypomethylation in gDNA.With different methylation-dependent restriction endonucleases, the aberrant demethylation was found to prefer C↓CGG sequences to CpG islands. The impacts of different dosage of Hcy showed that the varied detrimental effects of Hcy could be attributed to different concentrations via different mechanisms. In mild and moderate hyperhomocysteinemia, the Hcy may primarily influence the epigenetic regulation of gene expression through the interference of transferring methyl-group metabolism, while in more higher Hcy concentration, the notorious impacts may be more directly caused via oxidative stress, apoptosis, inflammation etc.

Objective To investigate the effect of small interfering RNA (siRNA) interference in the expression of epidermal growth factor receptor (EGFR) on the radiosensitivity of esophageal squamous carcinoma (Eca-109) and esophageal adenocarcinoma (OE-19) cell lines.Methods Human Eca-109 and OE-19 cell lines were selected as study subjects.Various EGFR-siRNA and negative siRNA were synthesized chemically through lipofection.Reverse transcription-polymerase chain reaction and Western blot were applied to measure the expression of EGFR before and after transfection,and the CCK8 assay was applied to analyze the influence of transfection on cell proliferation.Blank control groups of Eca-109 and OE-19 cells (O1 and O2 groups),simple irradiation groups (R1 and R2 groups),and EGFR-siRNA irradiation groups (E-R1 and E-R2 groups) were set,and the doses for single irradiation were 0,2,4,6,and 8 Gy.The colony-forming assay was applied to calculate survival fraction (SF) and sensitization enhancement ratio (SERD0 ratio),and flow cytometry was applied to evaluate the influence of EGFR-siRNA combined with radiotherapy on cell cycle distribution and apoptosis rate,and the dose for single irradiation was 6 Gy.Results The expression of EGFR in both cell lines was significantly down-regulated by EGFR-siRNA,and the inhibition rate of cell proliferation by transfection was＜5％ (4.9％ and 4.5％,respectively).The results of colony-forming assay showed that the cells in the E-R1 and E-R2 groups had a lower SF than those in the O1 and O2 groups,with an SERD0ratio of 1.40 and 1.01,respectively.The results from flow cytometry showed that compared with the E-R2 group,the E-R1 group had a higher proportion of cells in G2/M phase and a lower proportion of cells in S phase after irradiation (P=0.016 and 0.028),as well as a higher apoptosis rate (P=0.007).Conclusions Compared with the cell line OE-19,the cell line Eca-109 has a significantly increased radiosensitivity when treated with siRNA interference in EGFR expression.

Objective:To evaluate the feasibility of delineating subvolume target in radiotherapy for brain tumors using Gd-based contrast clearance difference.Methods:Twenty-six patients with malignant brain tumors were scanned with MRI. The first and second acquisitions of standard T 2-weighted images (T 2WI) and T 1-weighted images (T 1WI) were performed at 5 min and 60 min after injection of contrast agent. Delayed contrast extravasation (DCEM) MRI computed by Brainlab comprised regions of contrast agent clearance representing active tumors and regions of contrast accumulation representing non-tumor tissues. Based on T 2WI images, 14 patients with liquefaction necrosis were divided into group A, and 12 patients without liquefaction necrosis into group B, respectively. Then, gross target volume (GTV) was delineated on T 1WI images. Based on the GTV, active tumor (GTV tumor) and non-tumor regions (GTV non-tumor) were delineated on T 1WI-DCEM fusion images, while liquefaction necrosis (GTV liquefaction) and non-liquefaction (GTV non-liquefaction) were delineated on T 1-T 2WI fusion images. Finally, the differences between different subvolumes were compared by paired t-test. Results:In group A, the GTV non-liquefaction and GTV liquefaction were (13.65±18.15) cm 3 and (6.30±7.57) cm 3. The GTV tumor was (10.40±13.52) cm 3 and the GTV non-tumor was (9.55±14.57) cm 3. The GTV non-liquefaction was significantly increased by 16.3% on average compared with the GTV tumor ( P<0.05). The GTV non-tumor was significantly increased by 16.3% on average compared with the GTV liquefaction ( P<0.05). In group B, The GTV non-tumor was significantly reduced by 68.8% on average compared with the GTV tumor ( P<0.05). Conclusions:Compared with T 2WI, DCEM has advantages in identifying the liquefaction area and can clearly differentiate the subvolume of active tumors from non-liquefaction necrosis. DCEM provides evidence for guiding the delineation of subvolume in primary and metastatic brain tumors.

OBJECTIVE To control the spread of multi-drug resistant(MDR) bacteria in hospital and to ensure patients′ medical safety.METHODS We had established a network monitoring system,improved MDR monitoring,enhanced management of antibiotics rational use and cut off the transmission of MDR bacteria.RESULTS After taken these measures,the standard preventing of medical workers,handwashing compliance and specimen censorship rate had been greatly improved.Antimicrobial drug use was more standardized.In the last two years,we hadn′t found any epidemic outbreak due to MDR bacteria event.CONCLUSIONS Taking comprehensive prevention and control measures can prevent and control the development and proliferation of MDR bacteria and ensure patients′ medical safety.

With DQOL (diabetes quality of life) scale, quality of life was evaluated before and after diabetic education in 136 type 2 diabetic patients. The Cronbach′s ? of DQOL scale was from 0.809 to 0.849, suggesting that the DQOL scale did effectively reflect the life quality of type 2 diabetic patients.

Objective To evaluate the clinical effect after laparoscopic sacral colpopexy (LSC) of combined transabdominal-transvaginal approach on stage Ⅳpelvic organs prolapse (POP). Methods The clinical data of 65 patients undergoing LSC of combined transabdominal-transvaginal approach from January 1st, 2010 to July 30th, 2017 due to POP stage Ⅳ in First Affiliated Hospital of Guangzhou Medical University were retrospectively analyzed. Objective outcome was assessed by comparing preoperative and postoperative pelvic organ prolapse quantification (POP-Q) systems. Subjective effects were assessed by comparing pelvic floor distress inventory-short form 20 (PFDI-20), pelvic floor impact questionnaire short form (PFIQ-7), pelvic organ prolapse/urinary incontinence sexual questionnaire-12 (PISQ-12) and patient global impression of improvement (PGI-I). Results All 65 patients were successfully performed without any intraoperative complications. Fifty-three patients were followed in the clinic department and 12 were followed up by telephone. The follow-up duration was 6.1-80.3 months and the median follow-up duration was 24.5 months. The bleeding loss was 20-250 ml. Postoperative urethral catheter residence day was (2.5± 1.1) days, length of postoperative stay was (6.2±1.7) days. The postoperative POP-Q scores were compared with preoperative scores which had significantly improved except pb (all P<0.01). The objective cure rates of vaginal anterior wall, apical and posterior wall prolapse stageⅣwere 90% (47/52), 100% (23/23) and 95% (20/21).About PGI-I, except for 1 patient who chose"improvement", the other 64 patients (98%, 64/65) all chose"significant improvement". Furthermore, preoperative and postoperative PFDI-20, PFIQ-7, and PISQ-12 scores were all statistically significant (all P<0.01). Subjective efficacy was significant. Three cases (5%, 3/65) of postoperative fever occurred. Two cases (4%, 2/53) had mesh exposure. Six patients (11%, 6/53) had recurrence of postoperative prolapse. Five cases had recurrence of vaginal anterior wall prolapse and no reoperation was performed; 1 case was recurrence of posterior vaginal wall prolapse who diagnosed as vaginal posterior wall prolapse stage Ⅲ; no recurrence of apical prolapse. The rate of reoperation (including exposed-mesh removal and pelvic floor reconstruction surgery) was 5% (3/65). Conclusions The LSC of combined transabdominal-transvaginal approach has a high subjective efficacy rate. The objective cure rate in the case of apical prolapse stage Ⅳ is one hundred percent.The LSC of combined transabdominal-transvaginal approach has low mesh exposure, low postoperative infection and the reoperation rate, which is one of optional pelvic floor reconstruction surgery. However, there is still a risk of recurrence in patients with POP stageⅣwith severe bladder bulging.

Objective To investigate the effects of control status scale for diabetes (CSSD70), designed by diabetes center of Ruijin Hospital, in type 2 diabetes patients. Methods The correlations between CSSD70 and age, gender, duration of diabetes, educational backgrounds, insulin use, instruments for diabetes control, diabetes family history, symptoms at the onset of diabetes, complications, random plasma glucose and HbA_(1c) were analysed in 136 type 2 diabetes patients. The correlation between CSSD70 and another national questionnaires DQOL was analysed, and in order to determine the validity and stability of CSSD70, 30 patients were asked to complete the scales twice within one week. Results The coherent results in CSSD70 and DQOL were obtained, but CSSD70 was more comprehensive and sensitive. Almost all factors included in this study were related to CSSD70 total score and sub-scale score. The stability of CSSD70 was fine, and a close relationship between total score and sub-scale score was found. Conclusion The CSSD70 appears to be a good scale in evaluating the functional health status in type 2 diabetes, and is suitable for Chinese patients in evaluating diabetes control.

The relationships of therapeutic regime, glycemic control and quality of life with depression were observed in 193 type 2 diabetic patients. The results showed that 46.6% of the diabetic patients manifested depression in various degrees; the depression status was correlated with age or duration of diabetes; the patients under insulin treatment showed more marked depression than the other patients (P

OBJECTIVETo investigate the gene frequencies and mutation patterns of alpha thalassemia (alpha-thal) and beta thalassemia (beta-thal) in Liuzhou city of Guangxi Zhuang Autonomous Region.

METHODSCluster sampling was used. A total of 1 028 of umbilical blood samples were collected for a prevalence study of alpha-thal and a total of 1 312 healthy young people when receiving pre-marriage consultation were recruited for a beta-thal prevalence survey. Individuals live in city or town area of Liuzhou. A complete blood count as well as hemoglobin electrophoresis analysis were done in all of samples for phenotyping of alpha and beta-thals. Those with Hb Bart's for alpha-thal indicator and those with both microcytosis (MCV < 85 fl) and elevated levels of Hb A(2) (>/=4.0%) for beta-thal were further studied by DNA analysis. PCR-based methodologies were used to characterize the mutation contributions of alpha and beta-thals. All the subjects were tested for the state of carrying beta-thala alleles for evaluating the situation of the compound heterozygotes of alpha-thal with beta-thal.

RESULTSOf 1 028 random samples of umbilical blood screened, 112 of subjects were defined to be the gene carriers of alpha-thal. The alpha-thal carrier rate was as high as 11.19% including 3 compound heterozygotes. Five well-known types of alpha-thal alleles were detected with gene contributions of 37.4% (--(SEA) deletion), 31.3% (-alpha(3.7) deletion), 17.4% (-alpha(4.2) deletion), 12.1% (alpha(CS)alpha mutation), and 0.9% (alpha(QS)alpha mutation), successively. Of the 1 312 adult specimens studied, 89 with beta-thal including 14 of the compound higher Hb F subjects were detected. All of the 89 phenotypic beta-thal carriers had the mutations in the beta-globin gene, making the overall prevalence 6.78%. The commonly seen three mutations, beta CD41 - 42 (-CTTT) frameshift, beta CD17 (T-A) nonsense mutation and beta-28 (A-G) promoter variation were accounted for 90% of the beta-thal alleles in Liuzhou. Of these beta-thal subjects, 16 (accounting for 18%) were found to be the compound heterozygosity for a beta-thal and an alpha-thal with 9 different types of gene defects with a detection rate 1.22%.

CONCLUSIONData from ecidation of alpha and beta-thal gene frequencies and mutation spectrum in Liuzhou city was useful for genetic counselling and prenatal diagnosis of this disease.

Adult ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Counseling ; Humans ; Male ; Prevalence ; alpha-Thalassemia ; epidemiology ; genetics ; beta-Thalassemia ; epidemiology ; genetics

To analyze and evaluate the knowledge of Chinese Guidelines of Diabetes Prevention and Treatment in Shanghai medical staff. 175 medical staff working in endocrinology or community health were enrolled and evaluated by a questionnaire of guidelines about the state of professional, training, and related knowledge. Only 16. 6% medical staffwere trained about the guidelines( 46. 67% from the general hospitals, 14. 75% from secod-level hospital and 7. 14% persons from the community hospitals, P＜0. 01 ). The total correct answer rate of the guidelines was 37. 36%. The correct rate of community hospitals was lower than others( P＜0. 05 ). The rate of doctors' was higher than nurses'( P＜0. 05 ). There were difference between doctors and nurses with the key point of diabetes care knowledge in different level hospitals. The effective method of clinical training in diabetes care should be explored. We still have to work hard to promote the effect of diabetes control and prevention. Effective training about the guidelines should be enhanced. The cooperation between general hospitals and community health institutions in diabetes prevention and treatment should be enhanced.