Background and purpose: Tumor vasculature is increasingly recognized as a target for cancer therapy. In recent years, a fusion protein consisting of the extra cellular domain of tissue factor (truncated tissue factor, tTF) was fused to the antibody selectively binding to tumor vasculature. Antibody-truncated tissue factor(Ab-tTF) fusion protein specifically induced thrombotic occlusion of tumor vessels resulting in tumor growth retardation or regression in some types of solid tumors. However, there were still some disadvantages in the above approach. We constructed and expressed that the (RGD)_3-tTF fusion protein with peptides arginine-glycine-aspartic acid (GRGDSP, abbr. RGD)as the carrier of tTF to explore whether it bad the capability of targeting to tumor vasculature in the colonic carcinoma model. Methods: The (RGD)_3-tTF fusion gene consisting of the tTF was fused to three series-wound peptides RGD. The (RGD)_3-tTF construct was expressed in Escherichia coil BL21(DE_3). The fusion protein was purified through Nickel affinity chromatography column. The activity of inducing blood coagulation was detected by clotting assay and coagulation factor X (FX) activation assay. The specific binding to integrins α_vβ_3 was analyzed by indirect enzyme linked immunosorbent assay (ELISA). All these were compared with the fusion protein RGD-tTE Colonic nude mice models were randomly divided into 3 groups (1 nude mice per group).Tumors were stained by the (RGD)_3-tTE RGD-tTF fusion protein and tTF which were labeled with Fluorescein Isothiocyanate(FITC). The location of the (RGD)_3-tTF fusion protein in the colonic carcinoma bearing nude mice tissue was analyzed by immunofluorescence assay. Results: The (RGD)_3-tTF fusion protein retained tissue factor thrombogenic activities. With increasing concentration, the clotting time was shortened correspondingly. Under the conditions of Ca~(2+), the clotting time was 9.96±0.56 min when the concentration was 6 μmol/L(P<0.01). The (RGD)_3-tTF fusion protein could activise F X above 6 μmol/L concentration, which was similar to RGD-tTF fusion (F=0.147, P>0.05). The ability of the (RGD)_3-tTF fusion protein binding specifically to integrins α_vβ_3 was stronger than that of the RGD-tTF fusion protein in the same concentration (F=164.81, P<0.01), which was apparently indicated by the A_(405nm) 1.25 and 0.95 when the concentration was 0.24 μmol/L. Immunofluorescence assay showed that the (RGD)_3-tTF fusion protein was assembling in the tumor vasculature of the colonic carcinoma bearing nude mice. Conclusion: The (RGD)_3-tTF fusion protein which retained tissue factor thrombogenic activities could bind specifically and efficiently to tumor vasculature in the colonic carcinoma bearing mice through binding to the tumor marker integrins α_vβ_3. It might be a promising foundation for further studies on the colon cancer molecular targeted therapy with tTF as an effective factor.

Objective To investigate the relationship between serum progesterone level at the day with human chorionic gonadotrophin (hCG) administration and pregnant outcome from in in-vitro fertilization-embryo transfer(IVF-ET). Methods From Mar. 2002 to Apr. 2007, 786 cycles with serum progesterone measurement on the day of hCG administration for final oocyte maturation in IVF were analyzed retrospectively in Reproductive Medicine Center in First Affiliated Hospital of Nanjing Medical University.All stimulations were down-regulated with gronadotrophin release hormone agonist (GnRH-a) in both long protocols and short protocols before gonadotrophin stimulation. When the thresholds of serum progesterone were set at 5.5, 6.0,6.5,7.0,7.5,8.0,8.5 and 9.0 nmol/L, respectively. If the level of progesterone was less than the thresholds, those patients were in lower progesterone group, on the contrary, more than the threshold value, those patients were in higher progesterone group. The laboratory results and the clinical outcomes between all patients at lower and higher progesterone group at different thresholds value were analyzed. Results The rate of normal fertilization, quality embryos, successful implantation, chemical pregnancy, clinical pregnancy and live birth did not exhibit remarkable difference between patients with higher and lower serum progesterone level at multiple thresholds on the day of hCG administration in the 786 cycles (P >0.05). However, when the thresholds of serum progesterone were at 8.5 and 9.0 nmol/L, early abortion rates of 27.3% (3/11) and 3/7 in higher progesterone group were significantly higher than 8.8% (26/297) and 8.6% (26/301) in lower progesterone group (P<0.05). And the total abortion rates of 3/7 in higher progesterone group were significantly higher than 11.0% (34/301) in lower progesterone group when the thresholds of serum progesterone were 9.0 nmol/L (P<0.05). Conclusions This study did not prove the correlationship between progesterone level at the clay with hCG administration and the probability of clinical pregnancy or live birth. However, early abortion rates or the total abortion rates were associated with higher progesterone level when the thresholds of serum progesterone were at 8.5 nmol/L or 9.0 nmoL/L.

Objective We aimed to (i) observe the therapeutic effect of Yiqi Wenyang Prescription (YQWYP, which consists of milkvetch root, tangshen, dried ginger, cassig twig, ephedra, biond magnolia flower-bud, Chinese magnoliavine fruit, earthworm, and liquorice root) on an allergic rhinitis (AR) murine model and the protective effect on the tight junction (TJ) key proteins zonula occludens-1 (ZO-1) and occludin and (ii) elucidate whether YQWYP exerts these effects by inhibiting autophagy in nasal mucosal cells.Methods Thirty C57BL/6 mice were randomly divided into six groups with 5 mice per group: (i) the blank group, (ii) the model group, (iii-v) the YQWYP low-dose, mid-dose, and high-dose groups(7, 14, 28 g/kg), and (vi) the cetirizine group(1.667 mg/kg). The model was established by intraperitoneal injection and nasal stimulation of ovalbumin. 1 hour before the nasal stimulation, mice in the blank group or in the model group were given normal saline by gavage, mice in the YQWYP groups and those in the cetirizine group were given the corresponding medicines solution by gavage. After 7 days, mice behavior was scored. After that, the mice were sacrificed and the nasal mucosa tissues were harvested. The inflammatory reaction of the nasal mucosa was observed by HE staining, the proliferation of goblet cells in the nasal mucosa was observed by PAS staining, the IgE content in nasal lavage fluid was determined by ELISA, the expression levels of ZO-1 and occludin were analyzed by immunohistochemistry, and the expression levels of LC3B, P62, and Beclin1 in the nasal mucosa of the blank group, model group, and YQWYP high-dose group were analyzed by Western blotting.Results After the successful establishment of the AR model, compared with the blank group, the symptom scores of nose scratching, sneezing, and runny nose in the model group were higher, while the total scores was significantly higher(P<0.01). In the model group, the nasal mucosal epithelium was disrupted and destroyed, a large number of inflammatory cells were infiltrated, goblet cells showed hyperplasia, mucus production was increased, mucosal swelling was obvious, the amount of IgE in nasal lavage fluid was increased (P<0.01). The protein expression levels of ZO-1 and occludin were decreased (P<0.05), the protein expression levels of LC3BⅡ/Ⅰ and Beclin1 were increased (P<0.05), P62 protein expression was decreased (P<0.05). After medication intervention, compared with the model group, in the YQWYP high-dose group and the cetirizine group, the total score were reduced (P<0.01), the swelling of the nasal mucosa was reduced, the infiltration of inflammatory cells was inhibited, the number of goblet cells was decreased, the amount of IgE in nasal lavage fluid was reduced(P<0.01), the protein expression of ZO-1 and occludin were increased (P<0.05), the protein levels of LC3BII/I and Beclin1 were decreased (P<0.05), P62 protein expression was increased (P<0.05). Conclusion YQWYP has a good therapeutic effect on AR mice, and its mechanism may be related to the promotion of nasal mucosal repair by inhibiting autophagy of nasal mucosal cells.

Objective To understand the current situation and explore the influencing factors of delay in seeking medical treatment for common symptoms of residents in the rural areas of Sichuan province. Methods In July 2019,multi-stage random sampling was carried out in Zigong city,Sichuan province,and the data were collected by face-to-face questionnaire interview.The residents who had lived at hometown for more than half a year in the past year and had seen a doctor in the most recent month were surveyed.Logistic regression was adopted to predict the influencing factors of delay in seeking medical treatment. Results A total of 342 subjects were enrolled,and the incidence of delay in seeking medical treatment was 13.45%(46/342).Compared with the young and middle-aged(<65 years)people,the elderly(≥65 years)people were more likely to have delay in seeking medical treatment (OR=2.187,95%CI=1.074-4.457,P=0.031).The rural residents who gave higher score of the overall quality of township health centers were less likely to have delay in seeking medical treatment (OR=0.854,95%CI=0.735-0.992,P=0.039). Conclusions The occurrence of delay in seeking medical treatment for common symptoms of rural residents in Sichuan province is low.Age and the overall quality evaluation of township health centers affect the occurrence of delay in medical treatment among the rural residents in Sichuan province.Efforts should be made to improve the awareness of disease prevention among the elderly in rural areas.The investment in health resources in township health centers should be increased to strengthen the introduction and training of talents.These measures can improve the health services in township health centers,guide residents to make timely use of health resources,and reduce the occurrence of delay in seeking medical treatment.
Middle Aged ; Aged ; Humans ; Surveys and Questionnaires ; Logistic Models ; Rural Population ; China/epidemiology*