Objective: To construct a recombinant adenovirus (abbreviated as ET-M9-PEX) containing MMP-9 signal peptide and noncatalytic carboxyl-terminal hemopexin domain of human MMP-2, and to use the constructed adenovirus as a drug bioreactor in vivo to enhance the expression of an anti-angiogenesis factor for treatment of tumor by a gene therapy strategy. Methods: Adenovirus vector containing M9-PEX gene was constructed by PCR and gene recombination, and was packaged and amplified in L293 cells to obtain ET-M9-PEX recombinant adenovirus with infective ability. The expression and secretion of PEX in ET-M9-PEX-infected cells were detected by Western-blotting and immunofluorescent staining. The inhibitory effect of ET-M9-PEX-conditioned medium on EC cells proliferation was detected by growth curve and its inhibitory effects on angiogenesis and tumor growth were determined by chicken chorioallantoic membrane (CAM) assay in vivo. Results:ET-M9-PEX was successfully constructed and the expression and secretion of PEX in ET-M9-PEX-infected cells were verified. The ET-M9-PEX conditioned medium significantly inhibited the proliferating rate of EC cells. The tumor weights from ET-M9-PEX-infected PG cells in CAM and gradeⅠvessel number were reduced by 57.57%(P

This paper summarized the concept, diagnostic criteria, clinical treatment, outcome and the current issues faced of minimalconscious state.

Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). Methods ALA incubation time, wavelength applied to irradiate, concentration of ALA incubated, irradiation fluence may modulate the effect of 5-aminolevulinic acid based Photodynamic Therapy (ALA-PDT).The high-pressure mercury lamps of 400W served as light source, the interference filter of 410nm, 432nm, 545nm, 577nm were used to select the specific wavelength. Fluorescence microscope was used to detect the fluorescence intensity and location of protoporphyrin IX (PpIX) endogenously produced by ALA. MTT assay was used to measure the survival of cell. Flow cytometry with ANNEXIN V FITC kit (contains annexin V FITC, binding buffer and PI) was used to detect the mode of cell death. Results ① 1mmol/L ALA incubated 1×105/mL HL60 cell line for 4 hours, the maximum fluorescence of ALA induced PpIX was detected in cytomembrane. ② Irradiated with 410nm for 14.4J/cm2 can result in the minimum survivability of HL60 cell. ③ The main mode of HL60 cell death caused by ALA-PDT is necrosis. Conclusion ALA for 1mmol/L, 4 hours for dark incubation time, 410nm for irradiation wavelength, 14.4J/cm2 for irradiation fluence were the optimal parameters to selectively eliminate promyelocytic leukemia cell HL60 by ALA based PDT. The photosensitization of ALA based PDT caused the necrosis of HL60 cell, so it could be used for inactivation of certain leukemia cells.

Background:Gastric ischemia-reperfusion injury often leads to calcium overload,excessive free radical production, leukocyte infiltration and microcirculation disturbance.Post hypoxic treatment can effectively reduce the injury induced by hypoxia/reoxygenation (H/R).Galanin receptor 2 (GalR2)is distributed mainly in the digestive and nervous system, which can regulate many endocrine activity.However,the protective effect of GalR2 on human gastric epithelial cells injury induced by H/R is not clarified.Aims:To investigate the protective effect and its mechanism of GalR2 agonist post-conditioning on human gastric epithelial cells injury induced by H/R.Methods:H/R model was constructed on human gastric epithelial cells GES-1.Normal control group (N group),H/R group,M1145 (GalR2 agonist)treatment group (M group),SB203580 (p38MAPK inhibitor) +M1145 treatment group (S +Mgroup)and DMSO solvent control group (D group)were established.Survival rate of cells was measured by MTT assay.Apoptosis rate of cells was determined by flow cytometry,and cell apoptosis was examined by Hoechst staining.Level of lactate dehydrogenase (LDH)was measured by ELISA.Expressions of Bcl-2,Bax and p38MAPK were determined by real-time quantitative PCR.Results:Survival rate of cells was significantly lower in H/R group than that in N and M groups (P <0.05 ).Apoptosis rate of cells was significantly higher in H/R group than that in N,M and S +M groups (P <0.05 ),and apoptosis rate of cells was significantly lower in Mgroup than that in S +M group (P <0.05).Expression of LDH was significantly higher in H/R group than that in Mand S +Mgroups (P <0.05).Expression of Bcl-2 was significantly higher in N and M groups than that in H/R,S +Mand D groups (P <0.05);expression of Bax was significantly higher in H/R group than that in N,M and S +Mgroups (P <0.05);expression of p38MAPK was significantly lower in H/R and S +M groups than that in M group (P <0.05 ).Conclusions:GalR2 agonist M1145 plays an effective role in reducing the injury of GES-1 cells induced by H/R,the effect may be conducted through p38MAPK pathway.

Objective To evaluate clinical significance of pleural needle biopsy ( PNB) in diagnosis of pleural disease and its safety. Methods Totally, 594 cases with established pleural effusion, who were undergone PNB at least once in the First Hospital of Changzhou, Jiangsu province from June 1989 to September 2002, were studied retrospectively and analyzed. Results Sensitivity of PNB was 74% (302/ 408) for tuberculous pleurisy and 70. 2% (113/161) for malignant pleurisy. No characteristic pathological changes were found in PNB for the other 25 patients with pleural effusion due to other causes ( SLE, rheumatoid arthritis, leukemia, lymphoma, Castleman disease and Meigs syndrome) . Complications associated with PNB were pneumothorax (69. 0% ) , bleeding (31. 0% ) , fever ( 19. 0% ) and seeding of malignant cells (5. 0% ), and no special treatment was required for them, but one case of tension pneumothorax recovered uneventfully after 5 days of closed drainage. Conclusions PNB was a safe, simple and effective means of diagnosing tuberculous and malignant pleural effusion, and should be attempted extensively at grassroots hospitals. Seeding of malignant cells was uncommon, but should be kept in mind. Repetition of PNB could improve its sensitivity in diagnosis for malignant pleural effusion, with extremely few fatal complications.

Objective To study the expression and significance of vascular endothelial growth factor(VEGF) in serum of umbilical blood from the newborns of women with severe pre-eclampsia.Methods Radioimmunoassay was used to measure the serum VEGF of umbilical blood from 26 newborns of women with severe pre-eclampsia (severe pre-eclmpsia group) and 30 newborns of normal pregnancy women (control group). According to birth weight-gestational age relation,severe pre-eclampsia group included 15 newborns small for gestational age (SGA group) and 11 newborns appropriate for gestational age (AGA group).Results (1)The VEGF level of umbilical vein blood and umbilical artery blood in severe pre-eclmpsia group was significantly higher than that in control group respectively(t = 2.16, P < 0.05;t = 4.82, P < 0.01). (2) In control group and severe pre-eclampsia group, the VEGF level in umbilical artery blood was significantly higher than that in umbilical vein blood (control group: t = 2.24, P < 0.05;severe pre-eclampsia group: t = 3.00, P < 0.01). (3)In both control group and severe pre-eclampsia group, there was no significant correlation respectively between VEGF levels in umbilical vein blood or umbilical artery blood and newborns' gestational age or birth weight. (4)VEGF level of umbilical artery blood in SGA group was significantly higher than that in AGA group as well as in control group(t = 3.22, P < 0.01 ;t = 6.80, P < 0.01). Conclusions The serum VEGF level of umbilical vein blood and umbilical artery blood from the newborns of women with severe pre-eclampsia was significantly higher than that of normal pregnancy women respectively. The VEGF level of umbilical artery blood can reflect the severity degree of the damage of hypertensive disorder complicating pregnancy to their newborns to some degree.

Background and purpose: Human epididymis protein 4 (HE4) has been suggested to be a novel biomarker of epithelial ovarian cancer (EOC). The present study aimed to evaluate and compare HE4 with the commonly used marker, carbohydrate antigen 125 (CA125) and imaging examinations in detecting the recurrence of postoperative EOC. Methods:In this study, 92 patients with the recurrence of postoperative EOC were analyzed retro-spectively. The treatments method included the secondary operation (48 patients) and chemotherapy (44 patients). The sensitivity of CA125, HE4 and CT/MRI examinations in detecting the recurrence of postoperative EOC were analyzed and correlated with surgical pathology and clinical follow-up results. Results:The sensitivity of CA125 and HE4 were 58.7%and 61.9%with no signiifcant difference (P>0.05). The diagnostic sensitivity of CA125 combined with HE4 (80.4%) was statistically higher than that of 2 tumor marks being applied independently (P>0.05). The diagnostic sensitivity of CT/MRI examinations (88.0%) was statistically higher than that of CA125 and HE4 (P<0.05), and no signiifcant difference was observed between CT/MRI and combined CA125 and HE4 (P>0.05). The highest diagnostic sensitivity (97.8%) was attained by combining of CA125, HE4 and CT/MRI examinations, which was statistically high-er than that of CA125 combined with HE4 (P<0.01), and CT/MRI examinations (P<0.05). Conclusion:Serum HE4 is an effective tumor marker in detecting the recurrence of postoperative EOC as well as serum CA125. HE4 and CA125 may improve the diagnostic sensitivity statistically. The highest diagnostic sensitivity was attained by combining of CA125, HE4 and CT/MRI examinations, which is the suitable strategy in screening the patient of postoperative EOC.

Objective To investigate the risk factors for pregnancy-induced hypertension in Tangshan city.Methods Ninety-five pregnant women with hypertension were involved in questionnaire survey with contents ineluding general states of health and histories of past illness. Results Pregnancy-induced hypertension were more seen in winter, women aged above 35 years old with blood type A, bad educational background, bad nutritional status, hypertension history for family or herself, diabetes history and ere (P＜0.01 ). Conclusion There are ages, blood types, educational background, nutritional status, hypertension history, diabetes history for the risk factors for pregnancy-induced hypertension.

Objective To investigate the effects of commonly used inductive agents on peripheral blood monocytic myeloid-derived suppressor cells (M-MDSCs) in renal transplantation recipients and to discuss their possible mechanism.Methods The enrolled patients received rabbit anti-thymocyte globulin (rATG) or basiliximab for induction therapy,with the maintenance immunosuppressive regimen of tacrolimus,mycophenolate mofetil and steroid.The number of CD11 b + CD33 + HLA-DR-CD14 + CD1 5-M-MDSCs and cytokine levels in peripheral blood,including interferon-γ(IFN-γ),interleukin-2 (IL-2),IL-4 and IL-6,were measured by flow cytometry before and 1 week,2 weeks,1 month,2 months,3 months after operation.Results A total of 47 recipients (29 given rATG 29,and 18 given basiliximab) were included in this study.Compared to the patients with basiliximab,asignificant increase in the frequency of M-MDSCs was observed in the rATG group at 2nd month after operation (5.5％ ± 2.8％ vs.3.8％ ± 1.6％,P＜0.001) and at 3rd month after operation (7.0 ％ ± 3.1％vs.4.1％ ± 2.3 ％,P＜ 0.001),while there was no significant difference in the cell number between the two groups.In the cytokine detection,levels of IL-2 and IL-4 in the rATG-treated recipients were significantly higher at 2nd weekpostoperation (Pr2 =0.032,and PIL-4 =0.019)and 1st month postoperation (PIL-2 =0.024,PIL-4 ＜0.001) than the basiliximab group.Conclusions ATG promotes the expansion of M-MDSCs,which is associated with the secretion of IL-2 and IL-4 due to the lymphocytes depletion.The synergistic immunosuppressive effect may contribute to the induction of immune tolerance.

AIM: To investigate the plasma neurotensin(NT) concentration in patients with coronary heart disease: acute myocardial infarction(AMI),unstable angina(UA),stable angina pectoris(SAP), old myocardial infarction (OMI), and to study the relationship between the plasma NT level and the myocardial ischemia . METHODS: The plasma NT concentration of 30 patients with AMI,32 patients with UA,35 patients with SAP, 31 patients with OMI and 32 normal controls were determined by radioimmunoassay(RIA). RESULTS: The plasma NT level in patients with AMI(24 h),in patients with UA when angina attacked is significantly higher than that of healthy controls. The plasma NT level in patients with SAP, in patients with OMI is not significantly different from that of healthy controls. The NT level of patients with UA when angina attacked is significantly higher than that after 2 weeks treatment (P