Objective To evaluate the expression of Gadd45α and p38 MAPK in placentas and the correlations of Gadd45α protein and serum soluble vascular endothelial growth factor receptor-1 (sFlt-1) and soluble endoglin (sEng) in preeclampsia(PE). Methods Fifty-four pregnant women who delivered from September 2009 to March 2010 in the First Affiliated Hospital of Chongqing Medical University were chosen as the subjects. They were classified into mild preeclampsia group (n=20),severe preeclampsia group (n=16) and the control group (normal pregnant women underwent elective cesarean sections at term without labor and perinatal complications, n=18). Western blot and immunohistochemistry were employed to determine the expression and localization of Gadd45α and p-p38 MAPK protein respectively. Gadd45α mRNA level was determined by quantitative real-time PCR. The levels of seum sFlt-1 and sEng were measured by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and LSD-t test were applied for statistical analysis. Results (1)Immunohistochemistry identified that the positive stained cells were mostly located in trophoblast cells in normotensive placentas, whereas in preeclamptic placentas Gadd45α protein and p-p38 MAPK protein were detected in trophoblast and endothelial cells, as well as a few stromal cells at increased levels.(2)The mRNA levels of Gadd45α was significantly elevated in mild and severe preeclampsia groups compared to the control group (2.10±0.11 and 3.33±0.13 vs 1.01±0.18, P＜0.05), and Gadd45α mRNA level in severe group was significantly higher than in mild group (P＜0.05).(3)The data of Western blot revealed that the Gadd45α protein levels in each group were 0.22±0.11, 0.65±0.15 and 1.34±0.17, respectively, with significant differences between each group(P＜0.05). The p-p38 MAPK protein levels in each group were 0.32±0.08, 0.72±0.12 and 1.45±0.21, respectively, with significant differences between each group (P＜0.05). p38 MAPK protein levels in the total groups showed no difference(P＞0.05).(4)Compared with the control group, sFlt-1 and sEng concentrations in maternal circulation were significantly increasing in mild and severe preeclampsia groups, and concentrations in severe group were significantly higher than those in mild group (P＜0.05).(5) There were positive correlations between Gadd45α protein levels and the concentrations of serum sFlt-1 and sEng in each group( r=0.88 and 0.87, respectively all P＜0.05). Conclusions Upregulation of Gadd45α in preeclampsia placentas may play an important role in the pathogenesis of preeclampsia. It may induce the increased maternal serum levels of sFlt-l and sEng by activating p38 MAPK signaling pathway, leading to deficient cytotrophoblastic invasion and abnormal placental vascular reconstruction during pregnancy.

Objective To evaluate the expression and location of the growth arrest and DNA damage 45 alpha(Gadd45α)gene in human placenta and explore the relationship between Gadd45α and preeclampsia(PE).Methods Thirty-six women with preeclampsia who delivered from September 2009 to March 2010 in the First Affiliated Hospital of Chongqing Medical University were chosen as the study objects.They were classified into mild group( n = 20), severe group( n = 16) and elective term cesarean sections group without previous onset of labor and perinatal complications ( control group, n = 18 ).Placentas were collected after they delivered and immunohistochemical streptravidin-biotin peroxidase(SP) method and histoscore were employed to detect the expression and localization of Gadd45α protein.Gadd45α mRNA level was determined by reverse transcription polymerase chain reaction (RT-PCR) technique and western blot analysis was used to quantify Gadd45α protein expression level.Results (1)Gadd45α protein was detectable in placenta tissues of the mild and severe groups, mostly located in cytoplasm and nuclei of trophoblast, plus nuclei of vascular endothelial cells and a few stromal cells; whereas placenta tissues of control group showed weak staining, and only detectable in trophoblast, undetectable in vascular endothelial cells.(2)The Gadd45α mRNA levels in placenta tissues of the severe and mild groups were 0.75 ±0.07 and 0.44±0.13, respectively, significantly higher than that in control group (0.18 ±0.04, P ＜0.05);compared with mild group, Gadd45α mRNA level was significantly higher in severe group( P ＜ 0.05 ).(3)The Gadd45α protein levels in placenta tissues of the severe and mild groups was 1.34 ±0.17 and 0.65 ±0.15, respectively, compared with mild group, Gadd45α protein level was higher in severe group (P ＜0.05); they were both significantly higher than that in control group ( 0.22 ± 0.11, P ＜ 0.05 ).Conclusions Gadd45α mRNA and protein levels in placenta tissues of patients with preeclampsia are higher than that of normotensive women, and up-regulated with aggravation of preeclampsia; the Gadd45α protein is located in trophoblast cells, which are closely related to pathogenesis of preeclampsia.These results indicate that Gadd45α plays an important role in the pathogenesis and progression of preeclampsia.

OBJECTIVEThis study aimed to investigate the effects of different tooth preparations on the fracture strength and pattern of failure of teeth with severe wedge-shaped defect restored with post and core crowns.

METHODSAccording to whether the teeth above the wedge-shaped defect was removed (represented by B) or not (represented by A), the ferrule next to the wedge-shaped defect was prepared (represented by D) or not (represented by C), the cast post-and-core was chosen (represented by E) or glass-fiber post and resin core was chosen (represented by F). A total of 64 human mandibular premolar teeth were randomly divided into 8 groups: A1-1 (A + C + E), A1-2 (A + C + F), A2-1 (A + D + E), A2-2 (A + D + F), B1-1 (B + C + E), B1-2 (B + C + F), B2-1 (B + D + E), B2-2 (B + D + F), each group 8 teeth. All the teeth were prepared and restored accordingly and then mounted on an electronic pressure universal testing machine. The maximum fracture strength and the patterns of failure were recorded.

RESULTS1) The fracture strength of Group A1-1 > that of Group B1-1, Group A1-2 > Group B1-2, Group B2-1 > Group B1-1, and Group B2-1 > Group B2-2 with significant differences (P < 0.05). 2) The patterns of repairable fracture in Group A1-2 and B1-2 were both 37.5%, and that of the other groups were 0. Furthermore, the difference was significant, and Group A1-2 and B1-2 were higher than other groups.

CONCLUSIONThe maintenance of the overhang above the severe wedge-shaped defect aid in the improvement of the fracture strength of the tooth restored with post and core crown. The ferrule of the wedge-shaped defect is not recommended to be prepared. Furthermore, the glass-fiber post and resin core is favorable for the re-repair of the teeth than the cast post and core.

Objective To study expression and activation of p38 mitogen activated protein kinase (MAPK) in vascular endothelial cells dysfunction in preeclampsia. Methods From Sept. 2009 to Mar.2010, 54 pregnant women underwent deliveries in the First Affiliated Hospital of Chongqing Medical University were enrolled in this study, including 20 patients in mild preeclampsia group, 16 patients in severe preeclampsia group and 18 women with term cesarean section without perinatal complications as control group. Placental endothelial cells were labeled by CD34 to assay microvessel density (MVD) of each group. Immunohistochemical SP and western blot were used to detect localization and expression of p-p38 MAPK protein, respectively. The levels of sera soluble fms-like tyrosine kinase-1 (sFlt-1)and soluble endoglin(sEng) were measured by ELISA. Results ①The MVD of placenta were 103 ± 3 in control group, 81 ±5 in mild preeclampsia group and 63±4 in severe group, respectively, which showed statistical difference among each group (P＜0.05).②The expression of p38 MAPK protein were 0.84±0.05 in control group,0.90±0.14 in mild group and 0. 86 ±0.18 in severe group, which did not reach remarkable difference among each group (P＞0.05). The expression of p-p38 MAPK protein were 0.13±0.05 in control group,0.59±0.12 in mild group and 1.16±0.18 in severe group, which show statistical difference among each group(P＜0.05).(3) The localization of p-p38 was in trophoblast, endothelial cells and a few (5.2±0.3)and(10.9±0.4)μg/L in control group,(12.5±1.2) and (20.4±5.3)μg/L in mild group and (19.3±3.0) and (29. 5 ±3.7) μg/L in severe group. When drawing paired comparison in those p-p38 MAPK protein levels and the concentrations of serum sFlt-1, sEng in preeclampsia groups (r=0.68,P＜0.05;r=0.87,P＜0.05). Conclusions The remarkable activation of the p38 MAPK in the placenta of patients with preeclampsia induced the increased levels of sFlt-1 and sEng in maternal serum, which confer the injury of vascular endothelial cells that caused the significant decline of MVD in placentas. p38 MAPK signaling might be one of the key pathways in vascular endothelial cell dysfunction in preeclampsia.

Type 2 innate lymphoid cells ( ILC2s) are recently identified members of the innate lymphoid cell ( ILC) family. These cells are capable of producing Th2-type cytokines such as IL-5 and IL-13 in response to epithelial cell-derived cytokines IL-25 and IL-33 and play critical roles in allergic diseases such as bronchial asthma. Further investigations on ILC2s will enhance the better understanding of type 2 immune responses and may provide new strategies for the treatment of allergic asthma. In this review, we fo-cus on the origin, location and biological function of ILC2s as well as their possible roles in the pathogenesis of bronchial asthma.

Objective:To evaluate the effects of neuregulin-1 (NRG-1) pretreatment on diaphragmatic function in septic rats.Methods:Twenty-six clean-grade healthy adult male Sprague-Dawley rats, weighing 220-260 g, were divided into 3 groups using a random number table method: sham operation group (group S, n=6), sepsis group (group Sep, n=10) and NRG-1 group (group N, n=10). Sepsis was induced by cecal ligation and puncture in anesthetized rats.Group S underwent simple laparotomy.At 30 min before operation, recombinant human NRG-1β 10 μg/kg was injected through tail vein in group N, while the equal volume of normal saline was given in S and Sep groups.At 24 h after operation, the survived rats were sacrificed, and the diaphragm was removed for determination of the contractile function and contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay) and for examination of the pathological changes (by hematoxylin and eosin staining). Results:Compared with group S, the force-frequency curve was shifted downward, and the levels of TNF-α and IL-6 were increased ( P<0.01), inflammatory cell infiltration was observed in Sep and N groups.Compared with group Sep, the force-frequency curve was shifted upward , and the levels of TNF-α and IL-6 were decreased ( P<0.01), inflammatory cell infiltration was alleviated in group N. Conclusion:NRG-1 pretreatment can improve diaphragmatic contractile function in septic rats, and the mechanism is related to reducing diaphragmatic inflammatory responses.

Objective To develop a PCR-ELISA assay for the rapid, specific and sensitive detec-tion of human seasonal influenza virus ( H1, H3 and B) by using molecular biological and immunological methods in combination.Methods The primers were designed according to the genes encoding the matrix protein ( M) , the H1 and H3 hemagglutinin ( HA) of influenza A virus and the nonstructural proteins ( B-NS) of influenza B virus and then were labeled with biotin.The PCR products were detected by ELISA by use of an internal catching probe labeled with DIG.Results The minimum copy numbers of genes encoding the M, H1, H3 and B-NS proteins detected by the established assay were 1.43?103 , 8.67?102 , 3.86?103 and 5.45?103 copies/μl, respectively, which indicated that the PCR-ELISA assay was about 10 times more sensitive than agarose gel electrophoresis in the detection of PCR products.No cross-reactions between the different subtypes of influenza virus or different species of virus were observed.Moreover, a total of 104 clin-ical specimens of influenza virus were examined by the PCR-ELISA assay, the results of which were consist-ent with those of the virus isolation method.Conclusion The newly developed PCR-ELISA assay was a highly sensitive and specific method for the rapid detection and subtyping of influenza virus, suggesting the possibility of using it in laboratory for the surveillance and detection of influenza virus.

Objective The sensitivity and specificity of 18 FDG PET/CT are poor in the diagnosis of gastric cancer .Gastric signet ring cell carcinoma and Mucinous gastric carcinoma is known to have low fluorodeoxyglucose (18FDG) uptake,but not known for poorly differentiated gastric adenocarcinoma .This study was to investigate the value of 18 FDG PET/CT in the diagnosis of poorly differ-entiated gastric adenocarcinoma . Methods We retrospectively analyzed the results of 18 FDG PET/CT of 34 cases of histologically confirmed poorly differentiated gastric adenocarcinoma .We recorded the volume , location , and gastric wall invasion depth , and maxi-mum standardized uptake value ( SUVmax) of the tumors and analyzed the relationship of 18 FDG uptake with the clinicopathologic pa-rameters. Results By 18 FDG-PET/CT, poorly differentiated gastric adenocarcinoma was diagnosed in only 67.6% of the patients (23/34).SUVmax was found to be significantly correlated with age , gastric wall invasion, and tumor size (P<0.05), but not with gender , tumor location , tumor pathologic subtype , lymph node metastasis , and distant metastasis ( P>0 .05 ) .Logistic regression a-nalysis showed the tumor size to be the sole factor influencing the 18 FDG uptake of poorly differentiated gastric adenocarcinoma ( OR=0.37, 95%CI 0.154-0.920, P=0.03). Conclusion The di-agnostic value of 18 FDG-PET/CT is but limited for poorly differentia-ted gastric adenocarcinoma , and attention should be paid to its false-negative results .

Coding region instability determinant (CRD) is one of the influence factors of oncogene c-myc.Coding region instability determinant-binding protein (CRD-BP) can connect with CRD in order to protect CRD from nuclease attack,prevent rapid degradation of c-myc mRNA,and increase c-myc protein content.Beta-transducin repeats-containing protein (β-TrCP) can affect cell growth,differentiation,apoptosis and oncogenesis by regulating multiple signaling pathways and cell cycle.The overexpression of CRD-BP can upregulate the expression of β-TrCP and both of them play important roles in the tumorigenesis,progression,metastasis and invasion of colorectal cancer.

Objective To investigate the inhibition effects of an hTERT-promoter-dependent oncolytic adenovirus Ad-VT that expresses apoptin on human prostatic carcinoma cell PC-3. Methods MTT assay was used to measure viability of PC-3 cell which was infected by recombinant adenovirus.The viability was measured at time points of 12,24,36,48,60,72,84 and 96 h after infection.AO/EB staining,DAPI staining,Annexin V assay were used to investigate the lethal effect and style of Ad-VT on PC-3 cell in vitro.The Caspases were measured by whole cell extraction of PC-3 cells 48hrs after infection. Results Ad-VT,Ad-VP3 and Ad-GT inhibited the proliferation of PC-3 cell in vitro.Ad-VT and Ad-GT were more effective than Ad-VP3 on cell growth,P ＜ 0.05.At 48,72,96 h time points,the inhibition effect of Ad-VT on PC-3 cell exhibited a dose related manner.When infection at MOI 100,the inhibition effect of Ad-VT on PC-3 cells exhibited time related manner.The AO/EB staining,DAPI staining,Annexin V assay,Annexin V assays and Caspase assays showed that Ad-VT inhibited the proliferation of PC-3 cells by inducing apoptosis of prostate cancer cells,Loss of cytoplasmic membrane integrity. Conclusions The hTERT-promoterdependent oncolytic adenovirus Ad-VT could effectively suppress prostate cancer cells PC-3 growth.