Objective To investigate the changes of coagulation function in the neonates patients with gastrointestinal dysfunction and chnical significance. Methods Forty neonates with gastrointestinal dysfunction were included, which were divided into three groups according to diagnosis criterion of gastrointestinal dysfunction: early group, medium group and late group. Coagulation function was tested and neonatal critical illness score(NCIS) was done. Forty normal neonates were selected as control group. The difference of coagulation function among all groups was observed, and the relationship between coagulation function and NCIS were evaluated. Results Compared with control group, the indexs of coagulation function of early group was no statistical difference (P>0.05), however the result between medium group and late group was significantly difference (P<0.05). The difference of coagulation function between medium group and late group was also significant (P<0.05), the lower NCIS was, the more serious the gastrointestinal function was. Conclusions The more serious the gastrointestinal dysfunction is, the poorer the coagulation function is and the lower the NCIS is, which suggest coagulation function should be monitored in neonates with gastrointestinal dysfunction, and early intervention should be done accordingly.

Objective To investigate p16 promoter methylation in premature rats with chronic lung disease induced by hyperoxia. Methods Eighty premature Wistar rats were randomly divided into two groups: hyperoxia group (fraction of inspiratory oxygen) 0. 90 and control group (fraction of inspiratory oxygen 0. 21), 40 rats for each group. Semi-nested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction were applied respectively to detect p16 promoter methylation in lung tissues. Additionally, p16 mRNA and protein expressions in lung tissue were detected by reverse transcription- polymerase chain reaction, Western blot and immunohistochemistry method. Results The methylation was not found in control group by seminested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction, while was found in different aged rats of the hyperoxia group. The methylation detection rate was higher by using the semi-nested methylation-specific polymerase chain reaction (52.5%, 21/40) than that by methylation specific polymerase chain reaction (42.5%, 17/40) in the hyperoxia group,but there was no statistically significant difference between the two methods. The p16 mRNA in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21(1.73 ± 0.40 vs 2.11±0. 37,1.29±0. 19 vs 1.60±0. 27,0. 95±0.25 vs 1.72±0. 34, t=2.19, 2.95 and 10. 43,P＜0. 05). The p16 protein expressions by western blot in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21 also (88. 1±8. 7 vs 95.0±4.1,65.7±4.5 vs 83. 5±13.6 and 50.4±4.9 vs 86.7±11.9, t=2.27,3.95 and 13.40,P＜0.05). The expression of p16 mRNA (1.06±0.61) and protein (62.32±25.65) in lung tissues of rats with methylation was lower than that without methylation (1.63±0.62 and 94.93±22.21, respectively) (t=2.95, OR=0. 86;t=4.28, OR=0. 85,P＜0.01, respectively). Conclusions Exposure to hyperoxia might induce p16 promoter methylation in lung tissues in premature rats. Methylation risk increases as exposure time extends. p16 promoter methylation induced by hyperoxia might participate in the mechanism of lowering p16 mRNA and protein expression, but might not result in p16 gene silence.

Objective To investigate the effects of Par-4 gene silence on hydrogen peroxide-induced apoptosis in alveolar epithelial cells. Method The alveolar epithelial cells A549 were cultured and exposed to hydrogen peroxide. The siRNA sequences targeted Par-4 gene was chemically synthesized and transfected to A549 cells with or without the exposure of hydrogen peroxide. The cells were divided into normal control groups, hydrogen peroxide-treated group(The cells were treated with 0. 1 mmol/L hydrogen peroxide), hydrogen peroxide and Par-4-siRNA-treated group(The cells were treated with 0. 1 mmol/L hydrogen peroxide after transfection of Par-4-siRNA), Non-specific DNA sequence transfection control group. The apoptosis of A549 cells was quantified by flow cytometry. The expression of Smac protein was detected by Western blot.Electrophoretic mobility shift assay was applied for evaluating the change of E2F1 DNA binding activity. Relative activity of Caspase-3 was detected by clolorimetric assay. Results The percent of apoptotic cells in hydrogen peroxide and Par-4-siRNA-treated group was (29.7 ± 2.3) %, which was significantly lower than that of hydrogen peroxide-treated group [(54.2 ± 4.1)%, q= 8.91, P ＜ 0.01)]. Par-4 siRNA could significantly suppress the increase of Smac protein, E2F1 DNA binding activity and caspase-3 activity induced by hydrogen peroxide in A549 cells. Conclusions Par-4 gene silence induced by siRNA might inhibit the apoptosis of alveolar epithelial cells, which might be resulted from suppression of the up-regulation of Smac gene expression, E2F1 DNA binding activity and caspase-3 activity.

OBJECTIVE:To establish a RP-HPLC method for the determination of nitrochlodipine in rabbit plasma.METH-ODS:Chromatographic conditions and extractive conditions were optimized with orthogonal design,internal standard:felodipine,solid phase extract column:C18,column size:NOVA-PackC18(4.6mm?250mm),detection wavelength:237nm.RESULTS: The optimal chromatographic conditions were:mobile phase acetonitrile-water(55∶45),flow rate 1.0ml/min,column temperature 45℃.The extraction conditions were A1 B1 C3.The calibration curve was linear within the range of 6.25～400?g/L(r=0.9 996) and the minimum limit of detection was 2.5?g/L.The average RSD of within-day ranged from 3.81% to 5.58% and that of day-to-day from 4.60% to 8.76%.The average recovery ranged from 100.7% to 101.8%.CONCLUS-ION:The method is simple,rapid,highly accurate and precise.It can be used for the quantitative determination of nitrochlodipine in plasma.

Objective To discuss two simulation studies on the closed and open peritoneum in abdomenal operations. Meth-ods We selected 120 domestic female rabbits from animal laboratory of Qi Qi Ha’er Medical College at random and divided them into four groups(according to whether the peritoneum was open or not,the degree of peritoneum defect at the right side of incision and the existence of peritoneum hemorrhagic focus ),with 30 cases in each group. Group Ⅰ: no peritoneum suture and making a defect of 4cm?3cm at the right side of the incision; Group Ⅱ: no peritoneum suture and making a defect of 4cm?3cm at the right side of the incision,with a hemorrhagic focus at peritoneum defect;Group Ⅲ: with peritoneum suture; Group Ⅳ:with dense and compact peritoneum suture.And then we analyzed postoperative peritoneum healing progress. Results Observeations of the incision infection by the naked eye were that one case was identified in group Ⅰ,Ⅲ and Ⅳ;and a significant difference of occurrence rate was identified between the fallowing groups: groupⅠandⅡ(P

273 chinese have been studied with high voltage electrophoresis and immumofixation to survey the polymorphism of the third component of human complement in Shaanxi Provice Two C_3 phenotypes were found in this population, they are C_3SS and C_3FS The frequencies of C_3SS and C_3FS are 98.53% and 1.47% respectively. The frequency of C_3S gene is 0.9926 and C_3F gene 0.0073. These results are similar to other studies in China and this group was in Hardy-Weinberg equilibrium. Compared with Negro and Caucasian, the chinese population in Shaanxi Province has a significant difference in C_3 polymorphism.

Objective:To investigate the effect of homocysteine (Hcy) on peripheral vascular plaque formation in patients with type 2 diabetes mellitus (T2DM) combine subclinical hypothyroidism (SCH), and to provide guidance for clinical medication and prognosis judgement.Methods:A total of 125 patients with type 2 diabetes admitted to the NO.3201 Hospital from 2018 Jan to Dec 2019 were selected. 125 patients with type 2 diabetes mellitus were divided into T2DM without plaque group (40 cases), plaque group (23 cases), T2DM with SCH without plaque group (25 cases) and plaque group (37 cases) according to thyroid function and whether they had peripheral vascular plaque. Data were collected to analyze the influencing factors of peripheral vascular disease.Results:(1) The incidence of vascular plaque in T2DM group and T2DM with SCH group was 36.5%(23/63) and 59.7%(37/62), respectively, with significant difference ( P<0.05). (2) There were significant differences in serum thyroid stimulating hormone (TSH), apolipoprotein A (ApoA), triglyceride (TG) and Hcy between T2DM group and T2DM with SCH group ( P<0.05); there was significant difference in low-density lipoprotein (LDL-C) and Hcy between T2DM groups with or without plaque ( P<0.05); there were significant differences in ApoA and Hcy between T2DM with SCH group with or without plaque ( P<0.05). (3) Logistic regression analysis showed that Hcy was the risk factors for the occurrence of vascular plaque in T2DM and T2DM with SCH group ( OR=1.640, 2.695, P<0.05). (4) Receiver operating characteristic (ROC) curve showed that the area under the curve of Hcy in T2DM group was 0.842 and Youden index was 15.75 μmol/L; The Hcy's area under ROC curve was 0.945 and Youden index was 12.9 μmol/L in T2DM with SCH. Conclusions:Hcy is closely related to the presence of peripheral vascular plaque in T2DM patients with subclinical hypothyroidism. The detection of blood Hcy level can provide new ideas for early diagnosis and treatment of peripheral vascular lesions in T2DM patients with SCH.

Objective To amplify natural killer (NK) cells in vitro and explore its killing effect on ovarian cancer cells.Methods (1) The separation of NK cells and identification.A total of 20 ml peripheral blood of one healthy volunteer was collected in Nov.2015,Peking University People's Hospital.The peripheral blood mononuclear cells of normal volunteers were isolated,cultured in vitro and amplificated cultivation for 14 days with K562 cells transfected and expressing interleukin 21 (IL-21-K562) as nourish cells.The number and dynamic state of the growth cells were monitored during the cultured process.Cells were harvested and counted after 14 days cultured.The NK cells phenotypes were detected by flow cytometry.(2) The killing effect of NK cells on ovarian cancer cells:the ratio of effector cells (NK cells) and target cells (ovarian cancer cells and its control) was 50∶ 1,20∶ 1,10∶ 1,5∶1 or 1 ∶ 1,NK cells killing effect on ovarian cancer cells was detected by the lactate dehydrogenase (LDH) release experiments.Results (1) The results of NK cells establishment and phenotypic characterization:the cells were induced in vitro for 14 days by amplification culture.With the extension of incubation time,the number of NK cells increased constantly,from 2.0× 107 on day 0 to 5.1 × 109 on day 14.Obvious amplification of the total number of cells were detected for 255 times.Living cells unstained by trypan blue eventually reached 95％ above.Before and after the induction and amplification in vitro,the percentage of NK cells (CD3-CD56+cells) in CD3-cells were 2.33％ and 85.32％,respectively (P＜0.01),which covered the whole lymphocytes 1.06％ and 69.42％,respectively (P＜0.01),which showed that NK was the main cell type in the amplificated lymphocytes.(2) The killing rate of NK cells on ovarian cancer cells in vitro:the results detected by LDH release experiments showed that NK cells could performed strong nonspecific killing effect on ovarian cancer cell lines SKOV3,HOC1A,3AO and CAOV3,as well the normal ovarian cell line T29 and NK sensitive cell line K562,and the killing effect increased significantly along with the increase of effector cells and target cells ratio (P＜0.01).When the ratio was 1 ∶ 1,the killing rate was 37％ for K562,while the rate of killing of other cells was around 10％ (P＜0.05).When the effect-target ratio was 20∶1 and 50∶ 1,in addition to CAOV3 cells (more than 70％),NK cells had a kill rate of more than 80％ for other ovarian cancer cells lines and their control cell K562 and T29 cells (P＞0.05).Conclusion NK cells could be established in vitro and have a good non-specific killing effect on ovarian cancer cells.

OBJECTIVE To investigate the terminal bacterial pollution of hospital central oxygen supply system and the sanitizing effect of MX-1 disinfector. METHODS Timing many spot samples of terminal oxygen supply equipment (oxygen flowmeter, humid bottle, oxygen pipe) were quantified and cultivated through 24 hours continue supply. RESULTS Five cases of timing many spot samples testified that there were no bacteria oxygen pollution in the pipe oxygen supply in the Department of Respiratory Disease, the Second Affiliated Hospital of Dalian Medical University, and that there were no bacteria pollution in the terminal oxygen supply equipments if they were sterilized strictly, but there would be severe bacteria pollution if they were not sterilized strictly. This test found that lid of humid bottle and lapis were the main source of bacteria pollution. Connection with MX-1 disinfector in the terminal can avoid bacteria pollution. CONCLUSIONS Severe bacteria pollution will happen if the oxygen supply equipments aren't sterilized strictly; connection with MX-1 disinfector in the terminal can avoid bacteria pollution and ensure oxygen for cleanliness and safety.