Objective To analyze the clinical effect of the treatment of secretory otitis media with puncture combined with injection.MethodsThe control group was given tympanic membrane puncture combined with conventional drug treatment, and the experimental group was given combined with tympanic tympanic membrane puncture injection and conventional medicine for treatment.ResultsThe total effective rate of the experimental group (95.2%) was significantly lower than that of the control group (66.7%)(P<0.05);The incidence of complications in the experimental group (6.3%) was significantly lower than that in the control group(22.2%)(P<0.05).ConclusionClinical efficacy and safety of the treatment of secretory otitis media with puncture and injection.

Objective Detection of S-100B protein in different degree meconium pollution level in serum of children with value.MethodsIn 2012 June to 2013 December in our department were simple meconium stained amniotic fluid in term newborns in 73 cases, and set up for the observation group, and according to the amniotic fluid pollution degree is divided into 47 cases of amniotic fluid of Ⅰ-Ⅱdegree pollution group and 26 cases in grade Ⅲmeconium group during the same period, selected 20 cases without amniotic fluid contamination in term healthy newborns for the control group, the groups were compared in S-100B protein content difference.ResultsⅢmeconium stained amniotic fluid were within 6h serum S-100B was significantly higher than that in control group(P<0.01), and the degree of contamination of amniotic fluidⅠ-Ⅱgroup compared with the control group, no significant difference(P>0.05).Ⅲ meconium stained amniotic fluid in children with 72h also increased.ConclusionThird degree meconium stained amniotic fluid but normal Apgar score of newborns may still exist in clinical brain injury, so pay close attention to.

Objective:To explore the value of improved sequential organ failure assessment (ISOFA) in predicting the prognosis of Klebsiella pneumoniae sepsis. Methods:The clinical data of 379 patients with Klebsiella pneumoniae sepsis admitted to the First Hospital of Jilin University from January 1, 2018 to June 30, 2019 were retrospectively analyzed.They were divided into survival group and death group according to the 28-day prognosis, and the age, gender, C-reactive protein, procalcitonin, grade of bacterial resistance, and the occurrence of septic shock of the two groups were compared.Statistical analysis was conducted by using independent sample t test, Mann-Whitney U test and chi-square test. Risk factors for the prognosis of the disease was analyzed by logistic regression analysis.The correlation between ISOFA and other scoring system including Charlson′s weighted index of comorbidities (WIC), national early warning system (NEWS), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), simplified acute physiology scoreⅡ (SAPSⅡ), mortality in emergency department sepsis score (MEDS), sequential organ failure assessment (SOFA), acute gastrointestinal injury (AGI) was determined by Spearman correlation coefficient.The area under the receiver operating characteristic curve (AUROC) was used to compare the evaluation value of each scoring system for the prognosis of Klebsiella pneumoniae sepsis. The clinical significance of ISOFA risk stratification was evaluated by Kaplan-Meier survival curve. Results:Among the 379 patients with Klebsiella pneumoniae sepsis, 278 were in the survival group and 101 were in the death group.The differences of age, gender, the occurrence of septic shock, grade of bacterial resistance, C-reactive protein, procalcitonin, WIC, NEWS, APACHEⅡ, SAPSⅡ, MEDS, SOFA, AGI and ISOFA score between the two groups were all statistically significant ( t=-3.218; χ2=6.781, 24.374 and 27.208, respectively; Z=-3.689, -5.022, -4.396, -4.697, -7.438, -6.348, -6.358, -8.676, -6.680 and -11.658, respectively; all P<0.01). The calculation method of ISOFA was obtained by single factor analysis: ISOFA=SOFA+ 1.5×AGI.Multivariate logistic regression suggested that ISOFA, gender, SOFA, procalcitonin, C-reactive protein, and bacterial resistance were independent risk factors for the prognosis of the disease. The Spearman correlation coefficients of ISOFA and WIC, NEWS, APACHEⅡ, SAPSⅡ, MEDS and SOFA were 0.327, 0.371, 0.614, 0.564, 0.578 and 0.847, respectively. The AUROC and its 95% confidence interval for WIC, NEWS, MEDS, SAPSⅡ, APACHEⅡ, SOFA alone, SOFA plus AGI, and ISOFA to predict the prognosis of Klebsiella pneumoniae sepsis were 0.646 (0.584-0.708), 0.657 (0.597-0.716), 0.712 (0.654-0.771), 0.713 (0.653-0.773), 0.749 (0.693-0.806), 0.788 (0.737-0.838), 0.872 (0.826-0.917) and 0.891 (0.845-0.937), respectively.The results showed that ISOFA had the best predictive effect.The Kaplan-Meier survival curve suggested that there were statistical differences in survival rate among ISOFA low-risk level, medium-risk level, and high-risk level (all P<0.01). Conclusion:ISOFA has important clinical significance in predicting the prognosis of Klebsiella pneumoniae sepsis.

Objective to investigate the level of platelet leukocyte aggregates in patients with acute cerebral infarction and their short term prognosis.Methods 105 patients with acute cerebral infarction onset within 24 hours were selected continuously,then platelet leukocyte aggregates including neutrophil aggregates (PNA) and platelet monocyte aggregates (PMA) and platelet lymphocyte aggregates (PlyA) were detected by flow cytometry within 24 hours of admission and the incidence of 14 days.modified Rankin Scale(mRS) was performed at 14 days of onset,as a prognostic indicator,and the mRS score was good at 3.The score >3 mRS was divided into poor prognosis.The level of platelet leukocyte aggregates was detected in 50 healthy subjects.Results (1) The platelet leukocyte aggregates in patients with acute cerebral infarction group were significantly higher than that of the healthy group,which was statistically significant (P<0.05).(2)MRs score <3 group and mRS score >3 score comparison,the difference of white blood cell aggregates was statistically significant(P<0.05).Conclusion leukocyte aggregates could be used as an index of short-term prognosis in patients with acute cerebral infarction.

Objective To observe the effects of different doses of Sappan Lignum and Chuanxiong Rhizoma on tumor stem cells marker ABCG2 in vivo. Methods Sphere cells obtained from serum culture were inoculated in nude mouse armpit, which were randomly divided into 5 groups: control group, Sappan Lignum high- and low-dose groups, and Chuanxiong Rhizoma high- and low-dose groups. Each medication group was given relevant medicine for gavage. 21 days later, inhibition tumor rate and ABCG2 protein and mRNA expression were detected with confocal microscope, Western blot, and RT-PCR. Results The sphere cells obtained from serum free culture had the abilities of cancer stem cells, such as proliferation, anti-aptosis and high expression of cancer stem cells markers. Chuanxiong Rhizoma high- and low-dose groups could inhibit tumor growth (P<0.05), and the inhibitory rate of Chuanxiong Rhizoma low-dose group was higher than the Chuanxiong Rhizoma high-dose group. Sappan Lignum high- and low-dose groups inhibited tumor growth without statistical significiance (P>0.05). Compared with the control group, Chuanxiong Rhizoma low-dose group could significantly inhibit the expression of resistant protein of ABCG2. Sappan Lignum high- and low-dose groups could not inhibit the protein expression of ABCG2. Each medication group up-regulated the mRNA expression of ABCG2 except for Chuanxiong Rhizoma low-dose group. Conclusion Low dose of Chuanxiong Rhizoma can inhibit the expression of ABCG2 protein levels, which can be the targeting killer for cancer stem cells.

BACKGROUND: Magnetic field can affect the growth and division of cancer cells both in vivo and in vitro, however, the effects on apoptosis of human liver cancer cells induced by magnetic field is still unclear.OBJECTIVE: To explore the inducing effect of extremely low fre quency (ELF) magnetic field on apoptosis of human liver cancer cell SK-HEP- 1.JESIGN: An open experiment with cells as the observational subjects.SETTING: Institute of Biotechnology, Shanghai Jiaotong University.MATERIALS: The experiment was carried out at the Institute of Biotechnology, Shanghai Jiaotong University from September 2004 to January 2005. The subject was human liver cancer cell line SK-HEP-1, purchased from cell bank of Chinese Academy of Science, Shanghai.METHODS: SK-HEP-1 cells were inoculated to T-flasks at the density of 2.0×107 cells L-1, and cultivated in the DMEM containing 0.1 volume fraction of heat-inactivated fetal bovine serum and 2 mmol/L L-glutamine. Exposure groups were exposed to 50 Hz, 20 mT magnetic field and the control groups were run concurrently under the same conditions with the exposed cultures but in a separate incubator which was free of magnetic field during 8-day culture process. The apoptosis of SK-HEP-1 cells were defined by DNA ladder assay, Hoechst 33258 staining and AO/EB staining respectively on day 8.MAIN OUTCOME MEASURES: ① DNA fragmentation pattern formation. ② The abnormal nucleus formation. ③ The percentage of apoptotic cells.RESULTS: ① Detection of internucleosomal DNA fragmentation by DNA ladder assay: After 8-day ELF magnetic field exposure, DNA fragmentation pattern was detected by DNA ladder assay, which was not observed in control groups (free of exposure). ② Fluorescence microscopy analysis of apoptosis by Hoechst 33258 staining: Hoechst 33258 staining was used to investigate the changes in the nucleus of cells, and many apoptotic bodies containing nuclear fragments were found in ELF magnetic field exposed cells, but just about none in untreated cells. At the same time, cytoplasmic shrinkage was observed in cells cultured under the exposure. And in some cells, even the cell membrane was unable to keep intact. ③ Fluorescence microscopy analysis of cell apoptosis by acridine orange/ethidium bromide (AO/EB) double staining: After ELF magnetic field exposure, the percentage of viable cells in exposure groups (9.2%) was rather low compared with the control groups (91.8%), and was accompanied with a high apoptotic rate (72.3%), while only 4.2% in control group. A large number of apoptotic cells were at the early stage of apoptosis. The rate of apoptotic cells (18.5%)after treated by magnetic field was higher than that of control group (4%). With the AO/EB double staining, control cells appeared to be round,intact and bright green while some of the exposed cells exhibited irregular cell morphology and condensed nucleus.CONCLUSION: Apoptosis of human liver cancer cell SK-HEP-1 could be induced by 50 Hz, 20 mT magnetic field in vitro.

AIM To develop a UPLC-MS/MS method for the simultaneous content determination of adenosine,salidroside,chlorogenic acid,calycosin-7-O-β-D-glycoside,specnuezhenide,calycosin,formononetin,quercetin,apigenin and astragaloside Ⅳ in Zhenqi Fuzheng Capsules (Ligustri lucidi Fructus and Astragali Radix).METHODS The analysis of methanol extract of this drug was performed on a 40 ℃ thermostatic Inertsutain C18 column (75 mm × 3.0 mm,2 μm),with the mobile phase comprising of methanol-acetonitrile-4 mmol/L ammonium acetate flowing at 0.5 mL/min in a gradient elution manner.RESULTS Ten constituents showed good linear relationships within their own ranges (r≥0.996 0),whose average recoveries were 95.1％-104.3％ with the RSDs of less than 4.20％.CONCLUSION This sensitive and specific method can be used for the rapid quality control of Zhenqi Fuzheng Capsules.

Objective To explore the role of specific AT rich sequence binding protein 1(SATB1) and wnt/β-catenin signaling pathways in the regulation of trophoblast invasion and its effect in the pathogenesis of preeclampsia. Methods From March 2013 to March 2014, 20 cases of human villous tissues (early pregnancy group) from women of 8-10 gestational weeks who received artificial abortion at the First Affiliated Hospital of Chongqing Medical University, 18 cases of placental tissues (mid-pregnancy group) from women of 18-20 gestational weeks who had labor induction by water bag, 20 cases of placental tissues (normal full-term group) from healthy full-term pregnancy women and 20 cases of placental tissues (preeclamptic group) from women with preeclampsia who received elective c-section in were collected. Immunohistochemical SP method was utilized to determine the position of SATB1 and beta-catenin in villous tissues or placental tissues. Western blot was performed to analyze the expression level of SATB1 and beta-catenin in villous tissues or placental tissues. Immunofluorescence assay was used to determine the location of SATB1 andβ-catenin in HTR8/SVneo cells. Western blot was performed to detect the expression level of SATB1 and beta-catenin in HTR8/SVneo cells cultured in normoxia and hypoxia reoxygenation(H/R) condition. Co-Immunoprecipitation detection was used to evaluate the interaction between SATB1 andβ-catenin in placental tissues in preeclamptic group and HTR8/SVneo cells in H/R group. Gelatin zymography analysis was used to measure the activity of matrix metalloproteinases(MMP)-2 and 9 in placental tissues from preeclamptic group and HTR8/SVneo cells in H/R group. Results (1) In the normal full-term group, rare syncytiotrophoblastic nodule, less fibrinoid necrosis and abundant numbers of capillary could be observed in placental tissues. In comparison, there were obvious vacuolation in the cytoblast of syncytiotrophoblast, rich fibrinoid necrosis and poor numbers of villous capillary in placental tissues from preeclamptic group. (2) SATB1 could be found by immunochemical staining in placenta or villous tissues from all the groups. The staining intensity of SATB1 were more weakening in preeclamptic group than in the normal full-term group. (3) β-catenin could be found by immunochemical staining in placenta or villous tissues from all the groups. The staining intensity of β-catenin were more weakening in preeclamptic group than in the normal full-term group. (4) The protein expression levels of SATB1 in early pregnancy group, mid-pregnancy group, normal full-term group and preeclamptic group were 0.300 ± 0.009, 0.271 ± 0.015, 0.238 ± 0.018 and 0.153 ± 0.007, respectively. The protein levels of β-catenin among the above groups were 0.743±0.041, 0.648±0.021, 0.549±0.069 and 0.269±0.047, respectively. Both the expression of SATB1 andβ-catenin protein were significant decreased in placental tissues from preeclamptic group compared with the other three groups. (5) The SATB1 andβ-catenin protein was located in nucleus of trophoblast and a small amount was in the cytoplasm. The fluorescence intensity of both SATB1 and β-catenin in the H/R group were significantly decreasing when compared to the normoxia group. (6) HTR8/SVneo cells in H/R group showed a significant decrease in both SATB1 andβ-catenin protein levels when compared to the normoxia group. The protein level of SATB1 in the normoxia group was 0.213 ± 0.005, while was 0.083 ± 0.021 in the H/R group. The protein level ofβ-catenin in the normoxia group was 0.797±0.081, and was 0.543±0.131 in the H/R group. (7) There was an interaction between SATB1 and β-catenin in placental tissues from the preeclamptic group and HTR8/SVneo cells exposed by H/R. (8) The enzymatic activity of MMP-2 and MMP-9 protein were decreased significantly in placental tissues from the preeclamptic group (2.251±0.310, 1.447 ± 0.102, respectively) when compared to the normal full-term group (7.098 ± 0.451, 5.502 ± 0.197, respectively). MMP-2 and MMP-9 were significantly decreased in the H/R group (0.471 ± 0.104, 0.297 ± 0.103, respectively) when compared to the normoxia group (0.842 ± 0.209, 0.595 ± 0.100, respectively). Conclusion The expression of SATB1 decreased in the placenta of preeclampsia. This may influence the activity of MMP-2 and 9 by regulating Wnt/β-catenin signaling pathways, affect trophoblast invasion and eventually result in preeclampsia.

Objective To (e)xplore inhibition of endothelial progenitor cells (EPCs) against hepatic vein thrombosis after allogeneic bone marrow transplantation (BMT).Methods Balb/c mice were randomly divided into three groups: (1) BMT group [Balb/c mice were injected intravenously with 5 × 106 bone marrow cells after total body irradiation (TBI)]; (2) EPCs co-transfusion with bone marrow cells group: 5 × 105 EPCs were infused into recipient mice simultaneously; (3) Normal control group.Liver index was detected on the day 0,5,10,15 and 20 after transplantation.Hepatic vein thrombosis,hepatic cells and vascular endothelial damage were observed under the light microscopy after H&E staining.The injury of liver cells,liver veins,hepatic sinusoidal endothelial cells (SECs)and platelet adhesion conditions were observed under a transmission electron microscope (TEM).The proportion of activated platelets and TNF-α concentration in peripheral blood were detected by using flow cytometry.Results On the day 0,5,10,15 and 20 after transplantation,the proportion of activated platelets,liver index and TNF-α concentrations in BMT group and EPCs co-transfusion group showed an upward trend,peaked on the 15th day,and then decreased.However,they were still significantly higher than those in normal control group (P＜0.05).The above parameters in EPCs co-transfusion group at each time point were significantly lower than those in BMT group (P＜0.05).As compared with BMT group,platelet adhesion decreased,hepatic vein thromboses were reduced,hepatocyte swelling and necrosis were alleviated,and liver damage repaired rapidly in EPCs co-transfusion group.Conclusion EPCs co-transfusion with bone marrow cells could inhibit the hepatic veins thrombosis and ameliorate liver damage significantly.

Objective To establish a reproducible mouse model of hepatic veno-occlusive disease (HVOD) after allogeneic bone marrow transplantation (aallo-ABMT) and explore its pathogenesis.Methods Balb/c mice were randomly divided into three groups:(1) normal saline (NS) control group; (2) total body irradiation (TBI) group; (3) allogeneic bone marrow transplantation (allo-BMT) group.Liver weight,total bilirubin (TBil),tumor necrosis factor α (TNF-a),interleukin 6 (IL-6) and monocyte chemotactic protein 1 (MCP-1) were detected on the day 0,5,10,15 and 20 after transplantation.Hepatic vein and sinusoid congestion,infiltration of inflanmatory cells,and damage to hepatic cells and vascular endothelial cells were observed under the light microscopy after HE staining.Fibrosis of hepatic sinusoids and venule was observed under the light microscopy after Masson staining.Results Liver weight and TBil levels were elevated at 5th day and reached the peak at 15th day after all-ABMT.The changes of hepatic congestion and edema were obviously observed and there was infiltration of inflammatory cells at 5th and 10th day after alloABMT.At 15th and 20th day,hepatic congestion,edema and necrosis were reduced and liver damage was mainly presented with liver fibrosis and inflammatory infiltration.All mice died within 10 days after TBI,and hepatic congestion and edema were aggravated.As compared with NS control group,TNF-α,IL-6 and MCP-1 concentrations were significantly increased after all-ABMT.Conclusion A reproducible mouse model of hepatic veno-occlusive disease after all-ABMT was successfully established,and the pathogenesis was closely related to endothelial damage caused by total body irradiation,inflammatory cell infiltration and increased concentrations of cytokines.