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 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.

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 explore the diagnostic values of dual energy lung perfusion in the diagnosis of pulmonary embolism by using dual-source CT (DSCT). Methods Thirty patients with clinically suspected pulmonary embolism underwent dual-energy scanning with dual-source CT. The scanned data were integrated into three groups including 140, 80 kV and coefficient of 0.3. According to the CT pulmonary angiography (CTPA) of the fusion data, the patients were divided into pulmonary embolism group and normal group. The thin-slice reconstruction of data was analyzed using dual-energy perfusion imaging analysis software. The lung field was divided into upper, middle and lower part to make quantitative analysis of lung tissue perfusion. Paired t-tests were used in the normal patients to compare bilateral lungs, and independent samples t-tests were applied to compare the embolism group and normal group, while minimum intensity projection images (MinIP) were utilized in the assessment of lung ventilation. Results Dual energy CT showed symmetrical homogeneous perfusion in 16 normal cases, without significant perfusion defects. Quantitative analysis showed that left and right lung perfusion were (27 ± 7) and (28 ± 8 ) HU respectively, and no significant difference was found between the two sides ( t=-1.73, P ＞0.05 ).Perfusion of the left upper, middle and lower lung was ( 23 ± 6), (24 ± 6), and (28 ± 8) HU respectively, while the perfusion of right upper, middle and lower lung was (26 ±8), (27 ±8), and (28 ±9) HU respectively, showing no statistical significant difference between the two sides (t=-1.91, -1.96,-1.73 ,P＞0.05 ). Angiography of pulmonary embolism group(14 cases)showed filling defects in the pulmonary trunk, segments and sub-segments. Pulmonary perfusion imaging showed low perfusion or defectsin lung field that dominated by embolic vessels. Quantitative analysis showed that the perfusion of the whole lung and the middle and lower lung were (22 ±5), (22 ±8), and (21 ±8) HU in the embolism group,which were significantly different from the normal group (t=-2. 10, -2.32, -2.63, P＜0.05).Minimum intensity projection images showed a good consistency of abnormal ventilation zone area and perfusion abnormalities. Conclusions Pulmonary perfusion status, especially pulmonary embolism, can be analyzed by dual energy CT scanning. It helps to early discover and precisely locate the embolism.

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.

Objective To evaluate the accuracy of ultrasonographic measurement of the transverse diameter of the cricoid cartilage in selecting the cuffed endotracheal tube (ETT) size for pediatric patients.Methods A total of 120 pediatric patients of both sexes,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 1 month-6 yr,with body mass index of 10.9-31.2 kg/m2,undergoing endotracheal intubation and general anesthesia,were divided into group A and group B,with 60 pediatric patients in each group.The pediatric patients were intubated with a cuffed ETT in two groups.The ETT size was selected based on the transverse diameter of the cricoid cartilage measured by ultrasonography in group A.The ETT size was selected according to the age-based formula in group B.A tracheal leak was detected after intubation to determine whether or not the ETT size selected was appropriate.ETTs were replaced when the actually selected ones were not appropriate,and the number of replacement was recorded.The development of intubation-related complications was also recorded.Results The accurate rate of cuffed ETT size selected at the first time was 95％ in group A,and it was significantly higher than that in group B (60％) (P＜ 0.05).There was no significant difference in the incidence of intubation-related complications between the two groups (P＞0.05).Conclusion Uhrasonographic measurement of the transverse diameter of the cricoid cartilage produces higher accuracy in selecting the cuffed ETT size for pediatric patients and is worthy of clinical application.