Intrauterine growth retardation (IUGR) is one of the most commonly encountered developmental toxicity, which could lead to perinatal morbidity and mortality, be also extended from the fetus to adulthood, and seriously affect the quality of the population. Caffeine widely exists in a variety of daily beverages and some drugs. Its consumption is increasing year by year. Caffeine intake during pregnancy is one of the risk factors for IUGR. However, its mechanism of adverse outcome based on embryonic research is still unclear. In this paper, the possible mechanisms of caffeine-induced IUGR focusing on 3 important factors-the mother, placenta and fetus were explored. Caffeine's impact on the mother is the chronic activation of renin-angiotensin system; on the placenta, caffeine induces cell damage or the failure of the cell proliferation/apoptosis balance, leading to blockage of blood supply to the placenta; caffeine is also capable of directly affecting fetal development through interfering its neuroendocrine.

Objective To investigate the effects of high-frequency repetitive transcranial magnetic stimulation (rTMS) at different intensities on the ultrastructure of an ischemic brain penumbra and the expression of brainderived neurotrophic factor (BDNF) using rats with permanent middle cerebral artery occlusion (MCAO).Methods Forty-two rats were randomly divided into a blank control group,an MCAO model control group,a sham stimulation control group and an rTMS group.The rTMS group was divided further into 3 subgroups:an 80％ of motor threshold (MT) subgroup,a 100％ of MT subgroup and a 120％ of MT subgroup.The cerebral infarction model was established by right MCAO.rTMS treatment was given 24 hours after the MCAO model was successfully established.The rTMS group and sham stimulation control group were given 20 Hz rTMS with the planned intensities.The MCAO model control group was not given any stimulation.After 14 days of treatment,transmission electron microscopy,immunohistochemical and Western blotting ( WB ) methods were used to investigate the ultrastructure of the ischemic penumbra and the expression of BDNF.Results Damage reflected in the ultrastructure in the 3 rTMS subgroups was less than in the model control group and the sham stimulation control group.Expression of BDNF protein increased significantly in 100％ of the MT group and blank control group rats as compared with that in the sham stimulation control group,while the blank control group and the 3 rTMS subgroups had no statistically significant difference in comparison with the MCAO model control group.The expression of BDNF protein had no statistically significant difference between any of the groups.Conclusion 20 Hz rTMS might,especially at 100％ of the MT,promote the recovery of the ultrastructure of neural tissues in the ischemic penumbra after acute cerebral infarction and enhance the expression of BDNF in the ipsilesional hemisphere.This may be one of the important mechanisms of rTMS's effectiveness in the treatment of ischemic stroke.

The DNA demethylase genes are widespread in plants. Four DNA demethylase genes (LJDME1, LJDME2, LJDME3 and LJDME4) were obtained from transcriptome dataset of Lonicera japonica Thunb by using bioinformatics methods and the proteins' physicochemical properties they encoded were predicted. The phylogenetic tree showed that the four DNA demethylase genes and Arabidopsis thaliana DME had a close relationship. The result of gene expression model showed that four DNA demethylase genes were different between species. The expression levels of LJDME1 and LJDME2 were even more higher in Lonicera japonica var. chinensis than those in L. japonica. LJDME] and LJDME2 maybe regulate the active compounds of L. japonica. This study aims to lay a foundation for further understanding of the function of DNA demethylase genes in L. japonica.
Computational Biology ; DNA, Plant ; chemistry ; Genes, Plant ; Lonicera ; enzymology ; genetics ; Oxidoreductases, O-Demethylating ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Transcriptome

Objective To investigate the effects of 20 Hz repetitive transcranial magnetic stimulation (rTMS) with different intensities on neurobehavior and expression of glial fibrillary acidic protein (GFAP) in ischemic penumbra of rats with cerebral infarction,so as to explore the probable mechanism. Methods Forty-three rats were randomly divided into a blank control group( n =7 ),a model control group( n =7),a sham stimulation control group(n =8) and a rTMS group (n =21) ; the rTMS group was further subdivided into 3 subgroups:80％ MT subgroup,100％ MT subgroup and 120％ MT subgroup,with 7 rats in each subgroup.The cerebral infarction model was established by right middle cerebral artery occlusion (MCAO) in each group except the blank control group.The 3 rTMS subgroups were given 14 successive blocks of 20 Hz rTMS with corresponding intensity.The sham stimulation control group received sham treatment (without any output).The model control group was given no stimulation,and the blank control group did not receive any special treatment.Functional assessments were performed at 3 different time points.After 14-day treatment,the expression of GFAP proteins in ischemic penumbra were detected by immunohistochemistry technique. Results Functional outcome reflected from 3 behavioral tests in 100％ MT subgroup after 14-day stimulation was better than 1 day after operation,while in the other rTMS subgroups functional outcomes were just better in 2 behavioral tests.The expressions of GFAP in 3 rTMS subgroups were all less than that in model control group. Conclusions The 20 Hz rTMS with 80％ MT and 100％ MT might be safe and effective to improve the functional outcome in rats with acute cerebral infarction,especially 100％ MT.Decrease of expression of GFAP in ischemic penumbra might be one of the mechanisms of beneficial effects of rTMS in ischemia brain injury.

Objective To investigate the safety and feasibility of the reversed π-shaped esophagojejunal anastomosis in laparoscope-assisted total gastrectomy (LATG) for gastric cancer (GC).Methods The retrospective corss-sectional study was conducted.The clinicopathological data of 18 GC patients who were admitted to the Subei People's Hospital of Jiangsu Province between January 2015 and October 2016 were collected.All the 18 GC patients underwent LATG,surgical procedures included free stomach and lymph node dissection firstly,side-to-side jejunal anastomosis secondly and laparoscopic gastrointestinal reconstruction using reversed π-shaped anastomosis finally.Observation indicators:(1) surgical situations:side-to-side jejunal anastomosis method,conversion to open surgery,operation time,reversed π-shaped anastomosis time,volume of intraoperative blood loss and number of lymph node dissected;(2) postoperative situations:time for initial out-of-bed activity,time to initial anal exsufflation,time for postoperative water intake,time of drainage tube removal,postoperative complications and duration of hospital stay;(3) postoperative pathological examination;(4) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect diet intake,anastomosis patency,gastrointestinal obstruction and patients' survival up to March 2017.Measurement data with normal distribution were represented as (x)±s.Results (1) Surgical situations:18 patients underwent successful LATG and reversed π-shaped esophagojejunal anastomosis,without conversion to open surgery and perioperative death.Five patients used in vitro hand-sewn side-to-side esophagojejunal anastomosis through small incision of specimens sampling,and 13 completed all surgery under laparoscopy.Operation time,reversed π-shaped anastomosis time,volume of intraoperative blood loss and number of lymph node dissected of 18 patients were (187±12)minutes,(37±5) minutes,(735±18)mL and 29±2,respectively.(2) Postoperative situations:time for initial out-of-bed activity,time to initial anal exsufflation,time for postoperative water intake and time of drainage tube removal in 18 patients were (1.6±0.5) days,(2.3±0.4) days,(2.5±0.5) days and (7.5± 1.5) days,respectively.One patient complicated with esophagojejunal anastomosis fistula at postoperative day 3 was cured by drainage and symptomatic treatment and then discharged from hospital.Eighteen patients received regularly angiography using oral water-soluble contrast medium after recovering fluid diet intake,showing anastomosis patency and no contrast medium leakage,and then discharged from hospital.Duration of hospital stay of 18 patients was (12± 11) days.(3) Postoperative pathological examination:of 18 patients,15 were diagnosed with adenocarcinoma and 3 with signet-ring cell carcinoma.T2,T3 and T4 of T staging were respectively detected in 3,3 and 12 patients.N0,N1,N2 and N3 of N staging were respectively detected in 8,3,2 and 5 patients.Stage Ⅰ,Ⅱ and Ⅲ of TNM stage were detected in 3,5 and 10 patients,respectively.(4) Follow-up and survival situations:17 of 18 patients were followed up for 6-25 months,with a median time of 12 months.During the follow-up,2 patients were complicated with sour regurgitation and vomiting after eating at month 6 and 12 postoperatively and received gastrointestinal contrast examination,showing anastomotic stenosis,and then were cured by endoscopic dilation and discharged form hospital.Other patients had good diet and survival,without anastomotic complications.Conclusion The reversed π-shaped esophagojejunal anastomosis in LATG for GC is safe and feasible,with good short-term outcomes.

Objective To investigate the neuroimaging features in mitochondrial encephalomyopatbies with lactic acidosis and stroke-like episodes(MELAS).Methods Twenty-two clinically diagnosed patients who came from department of neurology,Huashan hospital in October 2003 to July 2006 were analyzed for CT,MRI,MRI contrast,MRA and MRS.Results In all 22 patients,the neuroimaging results of 21 were positive.There were 9 patients lying in hemisphere,12 in both cerebral hemispheres,including occipital,parietal,temporal and frontal lobe.The abnormal areas showed low signal intensity on T_1-weighted MRI,high signal intensity on T_2-weighted MRI and fluid attenuated inversion recovery(FLAIR)images.The lesions of 12/16 patients on MR contrasted images were enhanced.The lesions of one patient showed malacoma-like changes,one showed Fahr syndrome' s change and another showed high signal intensity on MR contrasted images.Conclusion Although the neuroimaging features of MELAS are complicated,the specific ones could help to make the diagnosis.

Objective To optimize the process conditions of one-step pelletization of Rouganbao Granules. Methods The factors influencing the pelletization of Rouganbao Granules were investigated by L9(34)orthogonal test, with the indexes of forming rate and fluidity. Results The spraying speed had the greatest effect on one-step pelletization, followed by atomization pressure and material temperature, and wind temperature had the smallest effect. At last, the best process parameters were relative density 1.15 (60 ℃), spray speed 55 mL/min, atomization pressure 0.25 MPa, wind temperature 75 ℃, material temperature 55 ℃, and critical relative humidity was 63%. Conclusion One-step pelletization technology can improve the preparation level and product quality, which can be used for the industrial production of Rouganbao Granules.

Objective:To investigate the effects of Clostridium butyricum on colitis and intestinal microbiota in mice with or without antibiotic pretreatment. Methods:Thirty specific pathogen free BALB/c mice were randomly divided into the blank control group, dextran sulfate sodium (DSS) group, antibiotic + DSS group, Clostridium butyricum + DSS group and antibiotic+ Clostridium butyricum + DSS group, with 6 mice in each group. After the mice were pretreated with quadruple antibiotics (ampicillin 1 g/L, neomycin 1 g/L, metronidazole 1 g/L, and vancomycin 0.5 g/L) in normal drinking water for 30 d, the mice colitis model was induced with DSS. At the same time, the mice in Clostridium butyricum + DSS group and antibiotics+ Clostridium butyricum + DSS group were given 1×10 6colony-forming unit (CFU) Clostridium butyricum by gavage. The effect of Clostridium butyricum on mice with colitis was evaluated by disease activity index (DAI), colon length and histopathological score. The level of serum inflammatory factors was detected by enxyme linked immunosorbent assay, and the effect of Clostridium butyricum on gut microbita in mice was determined by fecal 16S rRNA sequencing. Results:The general condition of mice of the blank control group were good, and their DAI scores fluctuated around 0. Since the fourth day after DSS drinking water was given, the mice of the DSS group showed signs of colitis such as weight loss, unformed stools and bloody stools. On the fourth day after intervention, the DAI score of Clostridium butyricum + DSS group was lower than that of DSS group (0.000±0.000 vs. 0.444±0.111), and the difference was statistically significant ( t=4.000, P=0.016 1). On the tenth and twelfth day after the intervention, the DAI scores of antibiotic+ Clostridium butyricum + DSS group were both lower than those of antibiotic+ DSS group (0.000±0.000 vs. 1.111±0.222, 0.667±0.000 vs. 1.889±0.222), and the differences were statistically significant ( t=5.000 and 5.500, both P<0.05). The histopathological score of mice colon tissue of Clostridium butyricum + DSS group was lower than that of DSS group (2.50±1.73 vs. 5.50±1.00), and the histopathological score of mice colon tissue of antibiotic+ Clostridium butyricum+ DSS group was lower than that of antibiotic+ DSS group (1.25±0.96 vs. 5.00±0.82), and the differences were statistically significant ( t=3.000 and 5.960, both P<0.05). The serum level of interleukin (IL)-1β Clostridium butyricum+ DSS group was higher than that of blank control group ((4.464±0.075) ng/L vs. (3.907±0.080) ng/L), the serum levels of tumor necrosis factor-α, IL-6 and IL-1β of Clostridium butyricum+ DSS group and antibiotic+ Clostridium butyricum + DSS group were all lower than those of DSS group ((2.402±0.383) ng/L , (1.845±0.345) ng/L vs. (6.958±1.084) ng/L, (1.752±0.146) ng/L, (1.307±0.048) ng/L vs. (3.537±0.608) ng/L, (4.464±0.075) ng/L, (4.066±0.190) ng/L vs. (7.477±0.339) ng/L), and the differences were statistically significant ( t=5.005, 3.964, 4.495, 4.693, 6.294, 8.674 and 8.774 , all P<0.05). The results of 16S rRNA sequencing showed that there were a significantly large number of anti-inflammatory or short-chain fatty acid producing bacteria in the gut microbiota of mice intervened by Clostridium butyricum, among which the dominant bacteria genus in Clostridium butyricum + DSS group and antibiotic+ Colstridium butyicum+ DSS group were Mucispirillum (linear discriminant analysis (LDA)=3.667 log10, P=0.004) and Stenotrophomonas (LDA=2.778 log10, P=0.044). In the antibiotic+ Clostridium butyricum+ DSS group, the dominant bacteria genus were Peptococcus (LDA=2.685 log10, P=0.018), Butyricimonas (LDA=2.712 log10, P=0.011), Bilophila (LDA=3.204 log10, P=0.014), Intestinimonas (LDA=3.346 log10, P=0.010), Candidatus- Saccharimonas (LDA=3.363 log10, P=0.029), Desulfovibrio (LDA=3.402 log10, P=0.025), Oscillibacter (LDA=2.870 log10, P=0.019) and Akkermansia (LDA=4.031 log10, P=0.005). Conclusions:Clostridium butyricum can effectively improve colitis in mice and regulate the intestinal microbial structure of mice, whlie antibiotic pretreatment can strengthen its regulation of intestinal microbiota to and enhance the efficacy of Clostridium butyricum.

OBJECTIVETo evaluate the effect of capsaicin on nude mice xenografted with colorectal carcinoma cells, and to explore its mechanism of action.

METHODSA nude mouse model of colorectal cancer was established by subcutaneous inoculation of human colorectal carcinoma HT-29 cells. Terminal deoxynucleotidyl transferase-mediated nicked labeling assay (TUNEL) was undertaken to detect the cell proliferation and apoptosis in the xenograft tissue in nude mice. Immunohistochemical (IHC) staining and Western blot were used to detect the expression of HSP27, Cyt-C and active caspase-3.

RESULTSThe tumor growth of the groups C10 and C20 was significantly slower than that of the group NS. The integrated optical density (IOD) of both the group C5 (2532.14 ± 578.11) and group C10 (6364.03 ± 1137.98) was significantly higher than that of the group NS (760.12 ± 238.05), (P < 0.05). The integrated optical density (IOD) of the group C20 was (15743.96 ± 1855.95), significantly higher than that of the groups C10, C5 and NS (all were P < 0.01). Immunohistochemistry showed that the cytoplasmic expression of HSP27 was strongly positive in the group NS, and significantly reduced with the increasing dose of capsaicin in the treated groups. The expression of active caspase-3 and Cyt-C in the group NS was weakly positive, and was significantly increased with the increasing dose of capsaicin in the groups C5 and C10 (P < 0.05), and the expression of active caspase-3 and Cyt-C of the group C20 was significantly higher than that of the groups C5, C10 and NS (P < 0.01). Western blot analysis showed that both the expressions of HSP27 of the group C5 (0.73 ± 0.05) and the group C10 (0.41 ± 0.03) were significantly lower than that of the group NS (P < 0.05). The expression of HSP27 of the group C20 (0.22 ± 0.06) was significantly lower than that of the groups C5, C10 and NS (P < 0.01). The expressions of active-caspase-3 and Cyt-C in the group C5 were (2.57 ± 0.34) and (2.03 ± 0.38), significantly higher than those of the group NS (P < 0.05). The expressions of active-caspase-3 and Cyt-C in the group C10 were (4.23 ± 0.45) and (3.13 ± 0.44), also significantly higher than those of the group NS (P < 0.05). The expressions of active-caspase-3 and Cyt-C in the group C20 were (5.78 ± 0.48) and (4.92 ± 0.52), significantly higher than those of the group C5, C10 and NS (P < 0.01). TUNEL analysis showed that there was a significant difference of cell apoptosis in comparison of each two groups. The higher dose of capsaicin was used, the more apoptosis was observed.

CONCLUSIONSCapsaicin can significantly inhibit the tumor growth and induce cell apoptosis in the colorectal carcinoma xenograft in nude mice. Its mechanism of action is possibly related with the down-regulation of HSP27 expression and up-regulation of expression of active caspase-3 and Cyt-C in the colorectal carcinoma xenograft in nude mice.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Capsaicin ; administration & dosage ; pharmacology ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Cytochrome c Group ; metabolism ; Dose-Response Relationship, Drug ; Female ; HSP27 Heat-Shock Proteins ; metabolism ; HT29 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Random Allocation ; Tumor Burden ; Xenograft Model Antitumor Assays

OBJECTIVETo evaluate the efficacy and safety of tenofovir disoproxil fumarate (TDF) in patients with chronic hepatitis B (CHB) after failure of nucleoside-analogues (NAs).

METHODSA total of 30 CHB patients who had been previously treated with NAs and had subsequently completed a 48-week course of TDF were retrospectively investigated. Patients' data of HBV DNA level (log10 copies/ml) and rate of undetectable HBV DNA at treatment weeks 0 (baseline), 4, 12, 24, 36 and 48 were collected for evaluation. The lower limit of HBV DNA detection was 100 IU/ml. The serum alanine aminotransferase (ALT) normalization rate, hepatitis B e antigen (HBeAg) seroconversion rate, viral breakthrough (VBT) rate, viral response (VR) rate, and adverse events were determined upon treatment completion. Statistical analyses were carried out using the Student's t-test, the x² test or the Kaplan-Meier method.

RESULTSOver the 48-week treatment period, HBV DNA levels declined significantly from baseline (week 4:(2.11 ± 0.38) log10 IU/ml, t =5.582; week 12:(0.93 ± 0.31) log10 IU/ ml, t =9.303; week 24:(0.75 ± 0.20) log10 IU/ml, t =3.123; week 36:(0.16 ± 0.19) log10 IU/ml, t =10.759; week 48:(0.14 ± 0.25) log10 IU/ml, t =12.202) (all P less than 0.01). However, the rates of HBV DNA reduction and of cumulative reduction were comparable at weeks 24, 36 and 48 (all P more than 0.05). The most robust decline in HBV DNA levels was observed at week 4 ((2.11 ± 0.38) log10 IU/ml) and the highest cumulative HBV DNA reduction was observed at week 24 ((3.79 ± 0.37) log10 IU/ml). The rate of undetectable HBV DNA at week 4 (26.7%) was significantly lower than that at weeks 24 (87.5%, P less than 0.01), 36 (80.0%, P=0.007), and 48 (88.9%, P=0.001). The median time to achieving undetectable HBV DNA was 10.4 weeks (range:3.43-34.0 weeks). At week 48, the rates of VR, HBeAg seroconversion, and VBT were 88.9% ,6.7%, and 0% respectively. During treatment, the levels of creatine kinase were more than two times the upper limit normal in 9.2% of the patients, and were comparable at each time point examined (all P more than 0.05). All patients showed a normal level of serum creatinine throughout the treatment period.

CONCLUSIONFor CHB patients with non-response to NAs, TDF can suppress HBV DNA replication very quickly and achieve a high rate of ALT normalization with a low rate of adverse events.

Adenine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; DNA, Viral ; blood ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Organophosphonates ; administration & dosage ; therapeutic use ; Retrospective Studies ; Tenofovir ; Young Adult