Objective To explore whether hypermethylation in the promoter of p16 gene and protein of p16 were associated with development and clinicopathological characteristics of colorectal cancer. Methods Methylation-specific PCR ( MSP) and immunohistochemistry SP were used to detected hypermethylation of p16 gene and p16 protein in tumor tissues from 32 patients with colorectal cancer. Results The hypermethylation of p16 gene was detected in 40. 6% of tumor tissues. The protein of p16 promoter was detected in 75% of tumor tissues. The hypermethylation of p16 gene was detected in 63% in Dukes stages of C and D tumors. The protein of p16 promoter was detected in 69% of tumor tissues. The hypermethylation of p16 gene was detected in 25% in the stages of A and B tumors. The protein of p16 promoter was detected in 81% of tumor tissues. The hpermethylation of p16 gene was detected in 100% in low differentiated carcinomas. The protein of p16 promoter was detected in 20% , the hypermethylation of p16 gene was detected in 30% , in the high and mediate differentiated carcinomas, the protein of p16 promoter was detected in 85%. Furthermore, the hypermethylation of p16 gene was detected in 63% in the lymph node metastasis and 25% in without lymph node metastasis. The protein of p16 promoter was detected in 65% in rectum and 100% in colon. Conclusions Our study demonstrated that p16 hypermethylation and protein were associated with the development of colorectal cancer and could be used as a putative prognostic indicator for this malignancy.

Objective To investigate the cerebral protective effect of anti-intercellular adhesion molecule-1 (anti-ICAM-1) antibody (1A29) in rats with global cerebral ischemia/reperfusion (I/R) injury. Methods Forty healthy SD rats weighing 180-200 g were randomly divided into 4 groups ( n = 10 each) : group Ⅰ received isotype-matched control antibody 1 mg/kg iv (control group); group Ⅰ ,Ⅲ ,Ⅳ received 1A29 1 mg/kg iv before cerebral ischemia (group Ⅱ), at the onset of reperfusion (group Ⅲ) and at 4 h of reperfusion (group Ⅳ) respectively. The global cerebral I/R was produced by 30 min four-vessel occlusion followed by 24 h reperfusion. All animals were killed at 24 h of reperfusion. Their brains were removed for microscopic examination and measurement of W/D weight ratio, infarct size and neurological deficits (0 = no deficit,4 = unable to crawl with mental depression) .Results The PMN leukocyte and monocyte infiltration, water content of the brain and infarct size were significantly decreased in group Ⅱ ,Ⅲ and Ⅳ as compared with control group. The neurological deficit scores were significantly lower in group Ⅱ ,Ⅲ, Ⅳ(1.6-1.8) than in control group (2.5).Conclusion 1A29 antibody administered either before ischemia or during the 4 h of reperfusion can protect the brain against global cerebral I/R injury.

AIM:To study the coating technique of pH-time lap colon-specific matrine delivery mini-pill consisted of the time lag release coating (inner layer) and enteric coating (out layer). METHODS:To filter the coating composition based on the index of dissolution of matrine and oxymatrine in vitro and the appearance rating of miui-pill. RESULTS:4 The coating composition of inner layer was the alcoholic solution,consisted of 2% EC,0.4% DEP and 2% talc powder. Then the coating composition of out layer was the alcoholic solution consisted of 5% Eudragit S100,3% talc powder and 0.5% TEC. The dissolution tests in vitro indicated that matrine and oxymatrine were not dissolved in the simulated gastric juice in 2 h. The accumulative amount of matrine and oxymatrine were less than 15% in the simulated intestinal fluid in 4 h. The amount of matrine and oxymatrine were 80.7% and 83.5% in the simulated colon juice in 2 h. CONCLUSION:The mini-pill could achieve the goal of delivering in the specific colon.

AIM:To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfide(DATS)in HL-60 cells.METHODS:HL-60 cells were either treated with various doses of DATS alone,or DATS combination with apocynin,a specific NADPH oxidase inhibitor,or with antioxidant N-acetyl-L-cysteine(NAC)for 0,1,3,6,12 and 24 h,respectively.The intracellular ROS level was measured by flow cytometry.The activity of NADPH oxidase was evaluated by NBT reduction experiment.The content of both malondialdehyde(MDA)and the protein carbonyl were analyzed by spectrophotometer.RESULTS:The results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells(P

Objective To explore changes of left ventricular filling during the strain phase of Valsalva maneuver (VM) and its mechanism. Methods Thirty healthy volunteers were recruited to perform VM with a load of 40 mmHg. Left ventricular filling parameters (E, A, E/A ratio, e and E/e ratio) were determined by echocardiography at baseline,at the first beat and at the second beat during the strain phase of VM,respectively. Results Compared to those at baseline, E, E/A ratio and E/e ratio increased ( P ＜0.05) while A and e did not change ( P＞0.05) at the first beat during the strain phase. Compared to those at the first beat during the strain phase,E, E/A ratio and E/e ratio decreased ( P ＜0.05) while A and e did not change ( P ＞0. 05) at the second beat during the strain phase. Conclusions Left ventricular filling decreased at the second beat during the strain phase of VM, which is different from the present knowledge that left ventricular filling would begin to decrease 4-5 beats later during the strain phase of VM. Positive intrathoracic pressure decreases left-side heart and pulmonary vessel' transmural pressure while increases the blood resistance, which may be the reason that E, E/A ratio and E/e ratio decreased at the second beat during the strain phase of VM.

Objective To explore the value of transthoracic echocardiographic suprasternal view in　the diagnosis of the subtype of the patent ductus arteriosus(PDA).Methods Sixty-five cases with PDA were examined by transthoracic echocardiographic suprasternal view and parasternal great artery short axis view respectively before closure therapy.The diameters of both the aotic and pulmonary side were detected,and subtype diagnoses were made.The results were compared with those from digital subtraction angiography (DSA).Results The demonstrated rates of PDA were 100％ on the parasternal great artery short axis and suprasternal views.Of the 65 cases,12 cases,19 cases and 19 cases were the funnel type of PDA checked on the parasternal great artery short axis view,suprasternal view and DSA respectively.The demonstrated rate of the parasternal great artery short axis view was lower than that of the suprasternal view (x2 =5.14,P ＜0.025 ).The diameter of the aotic side of PDA was (8.31 ± 2.76)mm,(10.87 ± 3.26) mm and (11.15±3.29)mm and the diameter of the pulmonary side of PDA was (5.69± 2.82)mm,(5.75 ± 2.63)mm and (6.09 ± 2.78) mm respectively on the above two views and DSA.Conclusions The diameter of the aotic side of PDA can be accurately detected by using superasternal view,which would be helpful for the diagnosis of PDA subtype.

Objective To investigate the role of metastatic lymph node ratio (MLR) in the evaluation of prognosis of patients with gastric cancer (GC) at different lymph node numbers examined.Methods Clinical data were reviewed retrospectively in a total 535 patients who underwent surgery for GC.Spearman correlation analysis between MLR or number of metastatic lymph nodes (N) and examined lymph node numbers,Kaplan-Meier method was used for comparison survival rates of N stage and MLR stage.A receiver operating characteristic (ROC) curve was used to evaluate the role of N stage and MLR stage in the prognosis of GC patients.Results Metastatic lymph node ratio and number of metastatic lymph nodes correlated with the examined lymph node numbers (r =0.146,r =0.378,P ＜ 0.01,P ＜ 0.001).The 5 year survival rate of MLR0,MLR1,MLR2 and MLR3 patients were 57.5％,69.9％,40.0％ and 21.7％ respectively when examined lymph node numbers ＜ 6 (P ＜ 0.01).The 5-YSR of MLR0,MLR1,MLR2 and MLR3 patients were 86.8％,59.2％,35.8％ and 39.2％ respectively when between 6-10 (P ＜0.001) and the 5-YSR of MLR0,MLR1,MLR2 and MLR3 patients were 88.7％,62.5％,0 and 17.7％ respectively when they ＞ 10(P ＜0.001).AUC of MLR staging was 0.68 ±0.05 when the numbers ＜ 6 (P ＜ 0.001).AUC of MLR staging was 0.72 ± 0.04 at numbers 6-10 (P ＜ 0.001).AUC of MLR staging was 0.79 ± 0.03 when numbers ＞ 10 (P ＜ 0.001).Conclusions MLR was less influenced by lymph node number examined than N.MLR stage has potential superiority to that the N stage in assessing prognosis of GC patients,especially for patients with more than 6 lymph nodes examined.

BACKGROUND:Human amniotic epithelial cells have some properties of stem cells, which can be induced to differentiate into corneal epithelial cells, but cannot be tracedin vitro. OBJECTIVE:To investigate the feasibility and infection efficiency of adenovirus vector carrying enhanced green fluorescent protein (EGFP) into the human amniotic epithelial cells. METHODS:The adenovirus vectors carrying EGFP was transferred into human amniotic epithelial cells culturedin vitro. After cultured and amplified, the morphology difference between transfected and non-transfected human amniotic epithelial cels was observed. The transfected human amniotic epithelial cells were observed under fluorescence microscope, and the cell cycle and the expression rate of EGFP in transfected human amniotic epithelial cells were detected by flow cytometry. RESULTS AND CONCLUSION:No obvious difference in the cell morphology was found between transfected human amniotic epithelial cells and normal human amniotic epithelial cells cultured in vitro. Flow cytometry analysis revealed that the EGFP positive rate was highest and reached up to 99.01% at 48 hours after transient transfection. The cell cycle of human amniotic epithelial cells transfected by the adenovirus vector was slowed a bit. To conclude, the adenovirus vector is a good medium of transfecting EGFP into human amniotic epithelial cells, and makes it more convenient to observe the further transformation of human amniotic epithelial celsin vitro.

Objective To evaluate the role of δ opioid receptor in the brain injury following asphyxial cardiac arrest-resuscitation in rats.Methods Ninety-six pathogen-free male Sprague-Dawley rats,weighing 300-350 g,were randomly divided into 4 groups (n =24 each):sham operation group (group S),asphyxial cardiac arrest-resuscitation group (group M),δ opioid receptor agonist BW373U86 group (group B) and δ opioid receptor antagonist naltrindole group (group N).Cardiac arrest was induced by clamping the tracheal tube for 8 min.Mechanical ventilation with pure oxygen was performed.Epinephrine 0.02 mg/kg and 5％ NaHCO3 1 mg/kg were injected intravenously as soon as chest compression was started.Appearance of spontaneous breathing and MAP ＞ 50 mm Hg (lasting for more than 10 min) were considered to be signs of successful recovery of spontaneous circulation (ROSC).BW373U86 and naltrindole 1 mg/kg were injected via the femoral vein immediately after ROSC in groups B and N,respectively,while the equal volume of normal saline was given instead in groups S and M.Neurological deficit score (NDS) was evaluated at 3,24 and 72 h after ROSC.The rats were then sacrificed,brains were isolated and the hippocampus was obtained for detection of the expression of brain-derived neurotrophic factor (BDNF)and tyrosine receptor kinase B (TrkB)mRNA by RT-PCR.The histological grading (HG) of neurons and number of survival neurons in hippocampal CA1 region were determined at 72 h after ROSC.Results Compared with group S,the expression of BDNF and TrkB mRNA was significantly up-regulated,HG was increased,and NDS and the number of survival neurons were decreased in groups M,B and N (P ＜ 0.05).Compared with group M,the expression of BDNF and TrkB mRNA was significantly up-regulated in group B,the expression of BDNF and TrkB mRNA was down-regulated in group N,and HG was significantly decreased,and NDS and the number of survival neurons were increased in groups B and N (P ＜ 0.05).NDS was significantly lower,the number of survival neurons was smaller,the expression of BDNF and TrkB mRNA was lower,and HG was higher in group N than in group B (P ＜ 0.05).Conclusion Activation of δ opioid receptor can reduce the brain injury following asphyxial cardiac arrest-resuscitation in rats and the mechanism may be related to up-regulation of BDNF and TrkB after activation of δ opioid receptor.

Objective:To explore the correlation between thyroid hormone content and treatment outcome in patients with biphasic disorder (BPD) and depressive disorder (DPD) before and after treatment.Methods:From June 2017 to June 2019, 72 patients with BPD(BPD group), 46 patients with DPD(DPD group) who met the diagnostic criteria of International Statistics and Classification of Disease and Health problems-10th Edition (ICD-10) in the Third People′s Hospital of Jiangmen, and 20 normal controls(control group) in our Hospital were enrolled in this study.The serum concentrations of total thyroxine (TT 4), total triiodothyronine (TT 3) and high sensitivity thyrotropin (HS-TSH) were measured at baseline, and the above hormone concentrations were measured in the study group after treatment for 12 weeks.The 17 Hamilton depression scale (HAMD-17) and Hamilton anxiety scale (HAMA), Bech-Rafaesdn Mania Rating Scale(BRMS) were used to evaluate the severity of symptoms at baseline and at the end of treatment.The curative effect was compared between the two groups, and the correlation between the curative effect (score reduction rate) and thyroid hormone level (concentration change) was observed. Results:At baseline, the concentrations of TT 4, TT 3 of patients with depressive phase in BPD group[(54.75±12.26)nmol/L, (86.26±15.29)nmol/L] were lower than those in the control group[(145.64±12.15)nmol/L, (156.78±36.54)nmol/L], and the TSH level in BPD group[(68.97±5.98)mIU/L]was higher than that in the control group[(45.59±9.28)mIU/L]; the levels of TT 4, TT 3 of patients with manic and mixed phase in BPD group[(166.75±12.25)nmol/L, (186.36±35.15)nmol/L] were higher than those in the control group, and the level of TSH in BPD group[(7.87±3.56)mIU/L] was lower than that in the control group; the concentrations of TT 4, TT 3 in DPD group[(65.59±23.11)nmol/L, (92.21±19.36)nmol/L] were lower than those in the control group, and the TSH level in DPD group[(69.38±14.75)mIU/L] was higher than that in the control group, the differences were ststistically significant ( t=5.867, 4.657, 2.369, 2.789, 3.247, 6.356, 4.358, 3.265, 2.365, all P<0.05). After treatment for 12 weeks, there were no statistically significant differences in TT 3, TT 4, HS-TSH concentrations among the three groups (all P>0.05). After treatment for 12 weeks, the scores of HAMa+ d, BRMS in BPD group and DPD group were significantly lower than those at baseline ( t=8.854, 12.321, 6.756, all P<0.05). The score of HAMa+ d was negatively correlated with TT 4, TT 3 concentrations, and positively correlated with HS-TSH concentration.The score of BRMS was positively correlated with TT 4, TT 3 concentrations, and negatively correlated with HS-TSH concentration. Conclusion:The level of thyroid hormone in patients with biphasic disorder and depressive disorder is different from that in normal subjects, and the level of thyroid hormone is closely related to the curative effect.