Objective To explore the influences of the final concentration and adding time of Cytochalasin-B (Cyt-B) on radiation-induced nucleoplasmic bridges (NPB) in cytokinesis-block assay.Methods Hunan peripheral blood samples were divided into 5 final concentration groups (group 2,4,6,8,10 μg/ml) according to different final concentrations of Cyt-B.Moreover,blood samples were divided into 4 adding time groups (group 0,28,40,44 h) according to different adding times of Cyt-B.Blood samples were irradiated with 0 (sham irradiation) and 2 Gy 60Co-rays in vitro,at a dose rate of 1 Gy/min.A cytokinesis-block assay was carried out to prepare NPB samples.The percentages of mononucleated,binucleated and multinucleated cells,as well as the frequencies of NPB and micronucleus (MN) in binucleated cells were analyzed using an optical microscope.Results Nuclear division index (NDI) and the percentages of binucleated cells increased with increased concentration of Cyt-B,and decreased with delayed adding time of Cyt-B (except group 0 h) in both final concentration groups and adding time groups.After exposed to 2 Gy,NPB frequencies were no significant difference (except group 0 h).MN frequencies had the trend of decreased with the increased concentration of Cyt-B,but no significant difference with adding time of Cyt-B.Conclusions In cytokinesis-block assay,different final concentration and adding time of Cyt-B may induce to the variation of NPB frequencies,but there was no significant difference.Appropriate increased final concentration or ahead adding time of Cyt-B can increase the percentage of binucleated cells that help to improve the efficiency of analysis.

Objective To observe the influence of the short effect antihypertension drugs- nifedipine and medial effect antihypertension drugs- extended release nifedipine on the blood pressure variability (BPV) in essential hypertension(EH). Methods Twenty-five EH patients were underwent 24-hour noninvasive ambulatory blood pressure monitoring (ABPM) and observed their BPV respectively before taking drugs, after taking nifedipine and extended release nifedipine. Meantime,25 normotensive controls (NC) were observed. Results (1)BPV in EH group was higher than that in controlled group and the severer the rise of blood pressure, the more obvious the increase of BPV (P 0.05). Conclusions Nifedipine could increase BPV but extended release nifedipine did not change BPV while they decreased blood pressure. Effect of extended release nifedipine was better than nifedipine in decreasing blood pressure.

BACKGROUND/AIMS: Chronic intestinal pseudo-obstruction (CIPO) is a serious, life-threatening motility disorder that is often related to bacterial overgrowth. Fecal microbiota transplantation (FMT) results in restoration of the normal intestinal microbial community structure. We investigated the efficacy of FMT in the treatment of CIPO patients. METHODS: Nine patients (age 18–53 years) with CIPO were enrolled in this prospective, open-label study. Patients received FMT for 6 consecutive days through nasojejunal (NJ) tubes and were followed up for 8 weeks after treatment. We evaluated the rate of clinical improvement and remission, feeding tolerance of enteral nutrition, and CT imaging scores of intestinal obstructions. Lactulose hydrogen breath tests were performed before FMT and 8 weeks after FMT to evaluate for the presence small intestinal bacterial overgrowth (SIBO). RESULTS: FMT significantly alleviated bloating symptoms, and symptoms of pain were relieved 2 weeks after FMT. Enteral nutrition administered through a NJ tube after FMT was well-tolerated by 66.7% (6/9) of patients. CT scores of intestinal obstructions were significantly reduced after FMT (P = 0.014). SIBO was eliminated in 71.0% (5/7) of patients. CONCLUSIONS: This pilot study demonstrated the safety of using FMT. FMT may relieve symptoms in selected patients with CIPO. FMT may also improve patient tolerance of enteral nutrition delivered via a NJ tube.
Breath Tests ; Enteral Nutrition ; Fecal Microbiota Transplantation* ; Humans ; Hydrogen ; Intestinal Obstruction ; Intestinal Pseudo-Obstruction* ; Lactulose ; Pilot Projects ; Prospective Studies

OBJECTIVETo evaluate the efficacy of periodic fecal microbiota transplantation (FMT) for refractory constipation.

METHODSClinical data of 49 patients with refractory constipation undergoing FMT through standard transplantation path of nasojejunal tube between April 2015 and April 2016 in Intestinal Microenvironment Treatment Centre of Nanjing General Hospital were analyzed retrospectively. Of 49 patients, 25 received single FMT for only 6 days (single group), and 24 received periodic FMT with another 6 days FMT 1 month after the first 6 days FMT (periodic group). The follow up was at 12 weeks after treatment. Autonomous defecation frequency, Wexner constipation score, gastrointestinal quality of life index and related adverse reaction were evaluated and compared at 4-, 8- and 12-week after treatment. Statistical analysis was performed on the difference after treatment at each time point, and the greater difference indicated the better improvement.

RESULTSThere were no statistically significant differences in general characteristics between the two groups (all P<0.05). Before treatment, Wexner constipation score was 17.32±2.66 and 16.25±2.47, gastrointestinal quality of life index was 81.84±8.73 and 83.25±7.87, autonomous defecation frequency was (1.64±0.57) time/week and (1.42±0.65) time/week in single group and periodic group respectively, whose differences were not significant (all P>0.05). Compared with before FMT treatment, the autonomous defecation frequency, Wexner constipation score, gastrointestinal quality of life index were obviously improved at the 4-, 8-, 12-week (all P=0.000). At the 4-week after FMT treatment, the improvement degree of autonomous defecation frequency, Wexner constipation score, gastrointestinal quality of life index was compared between two groups, and no statistically significant differences were found (all P>0.05). While at 8-week and 12-week after FMT treatment, as compared to single group, periodic group had greater Wexner constipation score (at 8-week: 7.29±2.05 vs. 5.96±2.30, t=2.135, P=0.038; at 12-week: 7.21±1.98 vs. 5.80±2.43, t=2.218, P=0.031), greater gastrointestinal quality of life index (at 8-week: 25.71±8.91 vs. 20.20±8.53, t=2.211, P=0.032; at 12-week: 24.16±8.99 vs. 18.92±8.28, t=2.127, P=0.039) and better autonomous defecation frequency [at 8-week: (2.42±0.93) time/week vs. (1.72±0.61) time/week, t=3.110, P=0.003; at 12-week: (1.37±0.88) time/week vs. (0.84±0.62) time/week, t=2.454, P=0.018].

CONCLUSIONPeriodic FMT has better efficacy than single FMT in the treatment of refractory constipation.

OBJECTIVETo evaluate the efficacy and safety of fecal microbiota transplantation (FMT) for gastrointestinal disorders.

METHODSRetrospective analysis of the clinical data of 406 patients who underwent FMT from May 2014 to April 2016 in the Intestinal Microenvironment Treatment Centre of Nanjing General Hospital was performed, including patients with constipation(276 cases), recurrent Clostridium Difficile infection (RCDI, 61 cases), ulcerative colitis(44 cases), irritable bowel syndrome (15 cases) and Crohn's disease(10 cases). Donors were completely unrelated, 18- to 50-year-old non-pregnant healthy adult, with healthy lifestyle and habits, without taking antibiotics, probiotics and other probiotics history within 3 months. There were three routes of FMT administration: patients received 6 days of frozen FMT by nasointestinal tube placed in the proximal jejunum under gastroscope (319 cases); patients received capsules FMT per day for 6 consecutive days (46 cases) or once 600 ml of treated fecal liquid infusion into colon and terminal ileum by colonoscopy(41 cases).

RESULTSClinical cure rate and improvement rate of different diseases receiving FMT were respectively as follows: RCDI was 85.2% (52/61) and 95.1%(58/61); constipation was 40.2%(111/276) and 67.4%(186/276); ulcerative colitis was 34.1%(15/44) and 68.2% (30/44); irritable bowel syndrome was 46.7% (7/15) and 73.3% (11/15) and Crohn disease was 30.0%(3/10) and 60.0%(6/10). RCDI had the best efficacy among these diseases(P<0.01). There was no significant difference between the three routes of FMT administration(P=0.829). The clinical cure rate and improvement rate of different routes were 43.3%(138/319) and 58.6% (187/319) respectively in nasogastric transplantation group, 41.5%(17/41) and 61.0%(25/41) in colonoscopy group, 37.0%(17/46) and 63.0% (29/46) in the capsule transplantation group. There was no serious adverse event during the follow-up. The most common side effects were respiratory discomfort (27.3%, 87/319) and increased venting (51.7%, 165/319) in nasogastric transplantation group. Diarrhea was the most common complication in colonoscopy group (36.6%, 15/41). The main symptoms were increased venting (50.0%, 23/46) and nausea(34.8%, 16/46) in oral capsule group. Side effect symptoms disappeared after the withdraw of nasogastric tube, or at the end of treatment, or during hospitalization for 1-3 days.

CONCLUSIONSFMT is effective for many gastrointestinal disorders. No significant adverse event is found, while the associated mechanism should be further explored.

Adult ; Clostridium Infections ; drug therapy ; Clostridium difficile ; drug effects ; Colitis, Ulcerative ; drug therapy ; Colonoscopy ; adverse effects ; methods ; Constipation ; drug therapy ; Crohn Disease ; drug therapy ; Diarrhea ; chemically induced ; Fecal Microbiota Transplantation ; methods ; statistics & numerical data ; Female ; Flatulence ; chemically induced ; Gastrointestinal Diseases ; drug therapy ; Gastroscopy ; methods ; Humans ; Intubation, Gastrointestinal ; adverse effects ; methods ; Irritable Bowel Syndrome ; drug therapy ; Male ; Middle Aged ; Nausea ; chemically induced ; Retrospective Studies ; Treatment Outcome

Objective:To screen radiosensitive lipid metabolites in rat plasma and analyze their metabolic pathways in order to provide scientific basis for radiation damage biomarker.Methods:The whole body irradiation of 60Co γ rays was performed to rats with different doses of 0, 1, 3 and 5 Gy. The changes of lipids in plasma were detected by untargeted lipidomics method based on liquid chromatography coupled mass spectrometry. Results:Twenty plasma lipids were identified as the potential radiosensitive biomarkers at 7 days after irradiation, including 13 over-expressed lipids and 7 down-expressed lipids, where 12 lipids well responded to radiation doses.Conclusions:Lipid metabolites in rat plasma are significantly changed after exposure to γ rays, and the metabolic pathways of sphingolipid, glycerophospholipid and glycosylphosphatidylinositol (GPI) are significantly enriched.

Objective:To investigate the metabolite changes in rat plasma after total body irradiation (TBI) and to explore dose classification based on radiation sensitive metabolites.Methods:The differential metabolites induced by radiation were screened and verified by metabolomics. In the discovery stage, 50 SD rats were irradiated with 0, 1, 2, 3, 5 and 8 Gy of 60Co γ-rays. In the verification stage, 25 rats were irradiated with 0, 0.5, 2.5, 4 and 6 Gy. Peripheral blood samples were collected 4 h after irradiation, and plasma was separated. Radiation-induced differential metabolites were identified and their concentrations were determined. Receiver operating characteristic (ROC) curve of the differential metabolites was used to classify dose range. Results:In the discovery stage, 8 radiation-induced differential metabolites in rat plasma were identified and four of them (cytosine, L-hexylcarnitine, Linoelaidylcarnitine and L-palmitylcarnitine) were upregulated, which was confirmed in the verification stage. The area under the curve (AUC) for the specific dose was >0.75. After combining these four metabolites, the AUC value to classify the radiation dose of 0 Gy versus >0 Gy, <2 Gy versus ≥2 Gy, <5 Gy versus ≥5 Gy were 0.96, 1 and 0.94, respectively.Conclusions:The metabolites in rat plasma changed significantly at 4 h after TBI, where 8 differential metabolites were identified. Cytosine, L-hexylcarnitine, linoelaidylcarnitine and L-palmiylcarnitine were stably over-expressed in the plasma after irradiation. The combination of these four compounds had high classification accuracy and thus may applicable as radiation sensitive biomarkers for dose classification.

Objective:To study the influence of circular RNA hsa_circZDHHC21_004 on the proliferation of human small intestinal epithelial cells HIEC-6 after 60Co γ-rays exposure. Methods:HIEC-6 cells were exposed to 60Co γ-rays at 0, 5, 10, and 15 Gy with a dose rate of 1 Gy/min. The expression level of hsa_circZDHHC21_004 in the irradiated HIEC-6 cell was detected. Hsa_circZDHHC21_004 was knocked-down to investigate the influences of hsa_circZDHHC21_004 on the proliferation of irradiated HIEC-6 cells by CCK-8 assay and colony formation assay. Results:The expression level of hsa_circZDHHC21_004 in HIEC-6 cells was upregulated by (1.00±0.24), (1.34±0.28), (1.85±0.31), and (2.80±0.64) times of control after 0, 5, 10, and 15 Gy irradiation, respectively and there were significant difference between 10 or 15 Gy group and 0 Gy group ( F=10.86, P=0.008). Knockdown of hsa_circZDHHC21_004 significantly increased the proliferation rate of HIEC-6 cells at 24, 48, and 72 h after 10 Gy irradiation compared with non-irradiated control ( t=-6.25, -5.83, -7.75, P < 0.001). Under 2 and 5 Gy irradiation, the clone formation rates of the hsa_circZDHHC21_004 knockdown cells were significantly higher than those of the control ( t=-7.45, -8.83, P<0.01). Conclusions:Hsa_circZDHHC21_004 is increased after irradiation and influenced the proliferation of irradiated HIEC-6 cells.

Objective:To screen the indicators of retrospective dose estimation, based on 5 cytogenetic methods to assess the victim followed-up at 4 year after 192Ir radiation accident in Nanjing. Methods:The chromosome aberration (dic + r) assay, cytokinesis block micronucleus (MN) and nucleoplasmic bridge (NPB) assay, fluorescence in situ hybridization (FISH)-based and G banding-based translocation analysis were used to retrospective biological dose estimation. Results:The estimated doses of FISH-based and G banding -based analysis were 1.45 and 1.21 Gy respectively, which was similar to the biological dose estimated short time after the accident. However, the estimated doses by chromosome aberration, micronucleus and nucleoplasmic bridge method were 0.56, 0.45 and 0.41 Gy respectively, which were lower than the corresponding biodose. Correction factors were used to adjust the biodose.Conclusions:In the 4th years after exposure, the estimated biological doses by FISH-based and G banding-based translocation were consistent with the biodose.Therefore, the two methods were suitable for retrospective dose estimation, while correction factors should be considered in chromosome aberration method for retrospective dose estimation.

Objective:To explore the effects of ultraviolet B (UVB) on the premature senescence of human immortalized keratinocytes HaCaT cells and the possible underlying molecular mechanism.Methods:HaCaT cells were exposed with UVB of different doses (20, 50, 80 and 100 mJ/cm 2). At 72 h after exposure, cellular morphology was observed by Giemsa staining, cell proliferation was detected by clone formation assay, and the proportion of premature senescence cells was detected by β-galactosidase staining. The number change of lysosomes was detected by Lyso-Tracker Red fluorescence probe at 24, 48 and 72 h after exposure. Cell migration was measured by scratch test at 24 h and 48 h after exposure. The protein expressions of p53 and p16 related to premature senescence were detected by Western blot assay at 72 h after exposure. Results:After UVB exposure, HaCaT cells showed a premature senescence phenotype. At 72 h after exposure, the cell volume increased ( F=115.18, P<0.05), the cell proliferation ability decreased ( F=410.32, P<0.05), the activity of β-galactosidase increased ( F=16.31, P<0.05), and the expressions of P53 and P16 increased. In addition, the number of lysosomes increased at 24, 48, and 72 h after exposure ( F=17.65, 38.36, 13.66, P<0.05), and cell migration capacity was inhibited at 24 and 48 h after exposure ( F=8.21, 11.48, P<0.05). Conclusions:UVB exposure can induce premature senescence of HaCaT cells by increasing the expression of p53 and p16 proteins.