Objective To design and synthesize targeting nuclear factor-?B (NF-?B) decoy-oligodeoxynucleotides (ODNs) and measure their anti-inflammatory actions. Methods Peritoneal macrophages ? (pM?) cells extracted from rats were randomly divided into normal control group, lipopolysaccharides (LPS)-stimulated group (Group A), decoy-ODNs treated group (Group B), non-decoy-ODNstreated group (Group C) and cationic liposome treated group (Group D). Supernatants and cells in the control group and the Group A were collected 1,2,6,12,18 and 24 hours after LPS stimulation. By using cationic liposomes in a ratio of 4:1 in quantity, pM? cells were transfected by decoy-ODNs at concentrations of 2 mg/L,4 mg/L and 8 mg/L respectively and stimulated with LPS 6 hours later. Then, supernatants and cells were collected 8, 12 and 18 hours after transfection. The expression changes and the distribution of p65 were analyzed by immunocytohistochemical method, the protein expressions of tumor necrosis factor ? (TNF?), interleukin-6 (IL-6) and interleukin-10 (IL-10) in the supernatants by enzyme-linked immunosorbent assay (ELISA) and mRNA transcriptions of TNF?,IL-6 and IL-10 by reverse transcriptase polymerase chain reaction (RT-PCR). Results Decoy-ODNs at concentration of 4 mg/L and 8 mg/L could markedly inhibit the expression and transcription of TNF? and IL-6 of pM? cells in a dose-dependent fashion but had a weak inhibitive effect on the expression and transcription of IL-10. Conclusions The targeting NF-?B Decoy-ODNs can suppress the expressions of inflammatory mediators in pM? cells.

Objective To analyze the clinical manifestations,diagnosis,treatment and prognosis of patients with splenic abscess.Method The clinical data,including baseline clinical data,clinical features,past history,pathogen culture result,treatment and the prognosis were retrospectively analyzed in the patients with the discharge diagnosis splenic abscess from January 1991 to March 2012 in Peking Union Medical College Hospital.Results The media time from onset to Peking Union Medical College Hospital of the 19 patients were 29 days.Among them,9 patients were cured,8 were improved and 2 died.Risk factors,such as tumor burden,diabetes,and using immunosuppressive agents etc,can be found in most patients with splenic abscess.All the 19 patients had splenic image changes and non-specific clinical features.The most common three clinical symptoms were fever(18 cases),chills (12 cases) and shivering (11 cases).The most common three signs were abdominal tenderness (9 cases),left upper quadrant sensitive to percussion (7 cases) and splenomegaly (4 cases).The most common etiological culture results were gram negative bacilli (9 cases),gram positive coccus (8 cases),and fungi (4 cases).Conclusions Clinical features are non-specific in splenic abscess patients.Related exam such as ultrasound should be performed on patients with splenic abscess risk factors to avoid misdiagnosis.Empiric antibiotic administration should begin right after the diagnosis based on the image.Pathogen culture should be timely conducted after pus collection.Individual therapeutical protocol should be chosen according to patient's condition.

Objective To construct a recombinant plasmid for expressing the HPV18 E2 gene and develop some materials for HPV18 related disease.Methods The E2 gene of HPV18 was amplified by PCR from PBR322-HPV18 and cloned into PIRES2-EGFP.The sequence of cloned HPV18E2 was confirmed by restriction analysis and DNA sequencing,then transfected into HeLa cells.E2 gene was detected by RT-PCR.Results HPV18 E2 gene fragment was amplified by PCR and ligated to PIRES2-EGFP,then transfected into HeLa cells successfully.The expression of green fluorescence cells was observed by fluorescence microscopy.Specific band could be seen in transfected cells by RT-PCR.Conclusion Recombinant plasmid PIRES2-EGFP is effectively expressed after being transfected into HeLa cells in vitro.

Objective To compare the biological characteristics of Culex pipiens pallens between permethrin-resistant strain and the susceptible strain. Methods In laboratory, the biological characteristics about bloodsucking, reproduction and development of permethrin-resistant and susceptible strains were observed and recorded. Life table of the experimental populations was constituted. Results The bloodsucking rate of resistant strain was lower than susceptible strain and the difference was significant. The life-span of adults of resistant strain and the developmental duration of eggs, larvae and pupae were longer than susceptible strain and the difference was significant. The death rate of eggs of resistant strain was higher than that of susceptible strain and the difference was significant. The permethrin-resistant strain showed a fitness value of 0 ^58 relative to the susceptible strain. Conclusion The permethrin resistance of Culex pipiens pallens may result in a disadvantage to its reproduction and development.

Objective To study the effect of reducing no-reflow after PCI treatment by early using Tirofiban and suction catheter in AMI patients .Methods 76 cases of patients were divided into group A (38 cases) and group B(38 cases) .The group A began to use Tirofiban with suction catheter to aspiration after coronary guidewire entering ,the suction were used in group B when the thrombus burden became exacerbation after balloon dilation .In addition ,chosen 38 cases of AMI patients treated with Tirofiban af-ter balloon dilation as group C .The influence of different treatment options to no-reflow and slow blood flow ,cardiovascular adverse events and the incidence of bleeding were observed .Results Group A compared with other two groups ,the no-reflow and slow flow rate had statistically significant differences (P< 0 .05) ,but there was no statistically significant differencebetween group B and group C(P>0 .05) .After three different surgical treatments ,the incidence of bleeding complications had no significant difference (P>0 .05 .The occurrence of adverse cardiovascular events had statistically significant between A group and C (P<0 .05) ,but there was no statistically significant difference between group B and group C (P<0 .05) .Conclusion Three kinds of treatment all have certain effect to reduce no-reflow in emergency PCI of AMI ,but early use of tirofiban with suction catheter in treatment of emergen-cy treatment has great clinical significance to reduce no-reflow .This study provides an effective treatment plan to reduce no-reflow in PCI for AMI .

Apoptosis of neutrophils controls the duration and the intensity of an inflammatory response and therefore the extent of neutrophil- mediated tissue damage, disturbance of neutrophil apoptosis has been associated with many diseases, underlying mechanism is not elucidated. C5a is a complement fragment that has multifunctional properties, which induces neutrophil chemoattraction, an oxidative burst, enhancement of phagocytosis, release of granule enzymes, and suppress neutrophil apoptosis. Several studies have reported calpain is involved in both neutrophil functions and apoptosis and it might play a more specific role in the regulation of neutrophil apoptosis. Diffenrent isoform of calpains is activted by diffenrent stimuli through different transduction pathway. It was reported previously that calpain is required for neutrophil migration and chemotaxis induced by C5a. In addition, autophagy is a ubiquitous physiological process that occurs in all eukaryotic cells and is considered to be a survival mechanism. Atg5 promotes autophagy and is indispensable to autophagosome formation. Upon calpain activation, Atg5 is cleaved and the resulting 24 ku Atg5 mediates apoptosis while losting the property of autophagy. Therefore, Atg5 represents a molecular switch between autophagy and apoptosis. The interaction among the C5a, calpain and Atg5 was introduced and new direction for further research was provided.

Objectiv e:To investigate the effect of different culture conditions on the differentiation of Treg and Th17 to lay a foundation for exploring the methods to reverse the immune tolerance induced by tumor microenvironment.Methods:The IL-6 gene was cloned and stablely transferred into the tumor cell line expressing TGF-β.The conditioned mediums ( CM) were prepared by collecting the culture supernatants of tumor cell lines with or without IL-6 expression and used in the in vitro culture of peripheral blood mononuclear cells ( PBMC ) .The changes of Treg and Th17 in PBMC treated with different CM were detected with flow cytometry ( FCM) .Results:The expression of TGF-βin BEL-7402 was higher than that in HepG2.Thus the BEL-7402 was selected for preparation of cell line stablely transfected with IL -6 gene.ELISA detection confirmed the effective expression of IL -6 by the identified cell lines.It was showed that the Treg increased in PBMC treated with culture supernatants of tumor cells .However,the presence of IL-6 reversed the increase of Treg and promoted the differentiation of Th 17.Conclusion: The culture supernatants of tumor cells increases the proportion of Treg.However,the presence of IL-6 in this CM can reverse the increase of Treg and raise the proportion of Th 17.

OBJECTIVETo explore the postburn adhesion properties of polymorphonuclear leukocyte (PMN) onto pulmonary vascular endothelial cells (PVEC) in burn patients with acute lung injury (ALI), so as to determine the role of C5a on PVEC-PMN adhesion.

METHODSMicrotubule sucking technique was employed to determine the PVEC-PMN adhesion. The myeloperoxidase (MPO) was also assayed to reflect the magnitude of PVEC-PMN adhesion.

RESULTSThe magnitude of PVEC-PMN adhesion increased and the adhesion force increased along with an increase in rh-C5a concentration. Simultaneously, the MPO activity was increased, which could be inhibited by anti-C5aR McAb in a concentration 1:104.

CONCLUSIONBoth C5a and C5aR participated in PVEC-PMN adhesion, which might be important in the pathogenesis of ALI.

Acute Disease ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Burns ; blood ; complications ; Cell Adhesion ; drug effects ; Cells, Cultured ; Complement C5a ; pharmacology ; Dose-Response Relationship, Drug ; Endothelium, Vascular ; cytology ; drug effects ; Fetus ; Humans ; Lung ; Lung Diseases ; complications ; Neutrophils ; cytology ; drug effects ; enzymology ; Peroxidase ; antagonists & inhibitors ; drug effects ; metabolism ; Receptor, Anaphylatoxin C5a ; Receptors, Complement ; immunology

Advanced paternal age has been overlooked, and its effect on fertility remains controversial. Previous studies have focused mainly on intracytoplasmic sperm injection (ICSI) cycles in men with oligozoospermia. However, few studies have reported on men with semen parameters within reference ranges. Therefore, we conducted a retrospective cohort study analyzing the reproductive outcomes of couples with non-male-factor infertility undergoing in vitro fertilization (IVF) cycles. In total, 381 cycles included were subgrouped according to paternal age (<35-year-old, 35-39-year-old, or ≥40-year-old), and maternal age was limited to under 35 years. Data on embryo quality and clinical outcomes were analyzed. The results showed that fertilization and high-quality embryo rates were not significantly different (all P > 0.05). The pregnancy rate was not significantly different in the 35-39-year-old group (42.0%; P > 0.05), but was significantly lower in the ≥40-year-old group (26.1%; P < 0.05) than that in the <35-year-old group (40.3%). Similarly, the implantation rate significantly decreased in the ≥40-year-old group (18.8%) compared with that in the <35-year-old group (31.1%) and 35-39-year-old group (30.0%) (both P < 0.05). The live birth rate (30.6%, 21.7%, and 19.6%) was not significantly different across the paternal age subgroups (<35-year-old, 35-39-year-old, and ≥40-year-old, respectively; all P > 0.05), but showed a declining trend. The miscarriage rate significantly increased in the 35-39-year-old group (44.8%) compared with that in the <35-year-old group (21.0%; P < 0.05). No abnormality in newborn birth weight was found. The results indicated that paternal age over 40 years is a key risk factor that influences the assisted reproductive technology success rate even with good semen parameters, although it has no impact on embryo development.
Pregnancy ; Infant, Newborn ; Female ; Humans ; Male ; Adult ; Paternal Age ; Retrospective Studies ; Semen ; Fertilization in Vitro ; Reproductive Techniques, Assisted ; Oligospermia

Objective To compare the differences and clinical value of prognostic evaluation between American Joint Committee on Cancer (AJCC) TNM staging system 7th edition and 8th edition for gastric cancer (GC).Methods The retrospective case-control study was conducted.The clinicopathological data of 1 383 GC patients who were admitted to the First People's Hospital of Changzhou between January 2008 and August 2012 were collected.Distal gastrectomy,proximal gastrectomy + pyloroplasty or total gastrectomy were performed according to preoperative evaluation and intraoperative exploration.Observation indicators:(1) surgical and postoperative situations;(2) follow-up and survival situations;(3) T staging comparison between AJCC TNM staging system 7th edition and 8th edition;(4) N staging comparison of AJCC TNM staging system 8th edition;(5) prognostic analysis in N staging of AJCC TNM staging system 8th edition;(6) TNM staging comparison between AJCC TNM staging system 7th edition and 8th edition;(7) prognostic analysis in different TNM staging between AJCC TNM staging system 7th edition and 8th edition.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to October 2017.Measurement data with normal distribution were represented as x ± s.Measurement data with skewed distribution were described as M (range).The survival curve and survival rate were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.Results (1) Surgical and postoperative situations:1 383 GC patients underwent successful radical gastrectomy,including 923 with distal gastrectomy,165 with proximal gastrectomy and 295 with total gastrectomy.Of 1 383 patients,115 with postoperative complications were improved by symptomatic treatment,including 87 with surgical complications and 28 with non-surgical complications.Postoperative pathological examinations:total number of intraoperative lymph node dissection and number of lymph node metastasis were 25± 12 and 7±4;577 didn't have lymph node metastasis and 806 had regional lymph node metastasis;308 were in early GC and 1 075 in advanced GC.(2) Follow-up and survival situations:1 383 patients were followed up for 1-117 months,with a median time of 34 months.The 1-,3-and 5-year survival rates of 1 383 patients were respectively 90.5％,71.9％ and 61.1％.(3) T staging comparison between AJCC TNM staging system 7th edition and 8th edition:T staging definition between AJCC TNM staging system 7th edition and 8th edition was identical.T staging of 1 383 patients:308,192,65,628 and 190 were respectively detected in T1,T2,T3,T4a and T4b stagings.(4) N staging comparison between AJCC TNM staging system 7th edition and 8th edition:N staging definition between AJCC TNM staging system 7th edition and 8th edition was identical.N staging of 1 383 patients:577,255,207,230 and 114 were respectively detected in N0,N1,N2,N3a and N3b stagings.N3a and N3b were classified as N3 staging of AJCC TNM staging system 7thedition,but they were classified as independent staging of AJCC TNM staging system 8th edition.(5) Prognostic analysis in N staging of AJCC TNM staging system 8th edition:5-year survival rate of patients in N0,N1,N2,N3a and N3b stagings was respectively 85.6％,76.5％,59.4％,45.2％ and 32.5％ based on AJCC TNM staging system 8th edition,with a statistically significant difference in survival (x2 =394.400,P＜0.05).There was a statistically significant difference between N0 and N 1 stagings (x2 =45.630,P＜0.05),between N 1 and N2 stagings (x2 =19.470,P＜0.05),between N2 and N3a stagings (x2 =7.602,P＜0.05) and between N3a and N3b stagings (x2=13.020,P＜0.05).(6) TNM staging comparison between AJCC TNM staging system 7th edition and 8th edition:TNM staging of 366 patients had changes,including 2 in T1N3b staging,2 in T2N3b staging,18 in T3N3b staging,120 in T4aN2 staging,149 in T4aN3a staging,34 in T4bN0 staging and 41 in T4bN2 staging;364 were detected in staging Ⅲ in 7th edition and 8th edition,and sub-staging of staging Ⅲ had a change;2 in T1N3b of ⅡB staging were redistricted into Ⅲ B staging based on AJCC TNM staging system 8th edition.(7) Prognostic analysis in different TNM staging between AJCC TNM staging system 7th edition and 8th edition:according to 7th edition,cases and 5-year survival rate were respectively 247,89.5％ in Ⅰ A staging and 147,83.7％ in Ⅰ B staging and 77,75.9％ in ⅡA staging and 207,70.5％ in ⅡB staging and 136,61.0％ in ⅢA staging and 236,37.5％ in Ⅲ B staging and 333,35.4％ in Ⅲ C staging,with a statistically significant difference in survival among sub-stagings (x2 =228.800,P＜0.05).There was a statistically significant difference in survival among Ⅰ,Ⅱ and Ⅲ stagings (x2=189.000,P＜0.05) and between ⅢA and ⅢB or ⅢC stagings (x2=22.710,18.010,P＜0.05).There was no statistically significant difference in survival between Ⅰ A and Ⅰ B stagings (x2=0.179,P＞0.05),between Ⅱ A and Ⅱ B stagings (x2 =0.265,P＞0.05),and between Ⅲ B and Ⅲ C stagings (x2 =1.550,P＞0.05).According to 8th edition,cases and 5-year survival rate were respectively 247,89.5％ in Ⅰ A staging and 147,83.7％ in Ⅰ B staging and 77,75.9％ in Ⅱ A staging and 205,70.7％ in Ⅱ B staging and 288,53.8％ in ⅢA staging and 258,37.3％ in ⅢB staging and 161,28.5％ in ⅢC staging,with a statistically significant difference in survival among sub-stagings (x2=234.900,P ＜ 0.05).There was no statistically significant difference in survival between Ⅰ A and Ⅰ B stagings (x2 =0.179,P＞0.05) and between Ⅱ A and ⅡB stagings (x2 =0.564,P＞0.05).There was statistically significant differences in survival between Ⅲ A and Ⅲ B or ⅢC stagings (x2 =29.790,43.060,P＜0.05) and between Ⅲ B and Ⅲ C stagings (x2 =7.494,P＜0.05).Further analysis showed that changes of TNM staging system between 7th edition and 8th edition were in T3N3b,T4aN2,T4aN3a,T4bN0 and T4bN2 stagings,5-year survival rate in above stagings was respectively 16.7％,35.8％,30.2％,47.1％ and 26.8％,with statistically significant differences in survival between T3N3b and T4aN2,T4aN3a,T4bN0 and T4bN2 stagings (x2 =19.590,8.039,12.070,3.853,P＜0.05),between T4aN2 and T4aN3a,T4bN2 stagings (x2 =6.529,3.859,P ＜ 0.05),between T4aN3a and T4bN0 stagings (x2 =10.400,P＜0.05) and between T4bN0 and T4bN2 stagings (x2=4.636,P＜0.05).There was no statistically significant difference in survival between T4aN2 and T4bN0 stagings (x2 =3.607,P＞0.05) and between T4aN3a and T4bN2 stagings (x2 =0.029,P＞0.05).Conclusions Compared with AJCC TNM staging system 7th edition,N3a and N3b stagings are classified as independent staging in AJCC TNM staging system 8th edition,and 8th edition is more accurate in prognostic evaluation of GC patients in stage Ⅲ.