1.Correlation of Apoptosis of Peripheral Blood Mononuclear Cell with Plasma 8-iso-prostaglandin F_(2?) in Neonates with Hyperbilirubinemia
dong, ZHOU ; xin, XIAO ; li-li, KANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To explore the correlation of the apoptosis of peripheral blood mononuclear cell (PBMC) with plasma 8-iso-prostaglandin F2? (8-iso-PGF2?) level in order to confirm the peroxidation damage of hyperbilirubinemia in neonates with jaundice.Me-thods One hundred and twenty-nine neonates who were 2 to 7 days old were divided into 4 groups:slight jaundice (43 cases),moderate jaundice (38 cases),serious jaundice (18 cases),and healthy control(30 cases)groups.Plasma 8-iso-PGF2? levels were assayed by enzyme immunoassay(EIA),and the apoptosis rates of PBMC were determined by flow cytometry.Results 1.There was no significant difference in the apoptosis rate of PBMC between normal and slight jaundice neonates (P=0.108);apoptosis rate of PBMC was increasing with the aggravation of jaundice.2.Plasma 8-iso-PGF2? level in normal neonates was 14.74?6.71 ng/L,there was no difference between normal neonates and neonates with slight jaundice(P=0.502).Plasma 8-iso-PGF2? levels in neonates with moderate and serious jaundice were much higher(Pa=0).3.Positive correlation existed between plasma 8-iso-PGF2? level and PBMC apoptosis rate (r=0.602 P=0).Conclusions Hyperbilirubinemia can induce peroxidation damage,resulting in increase of PBMC apoptosis.Plasma 8-iso-PGF2? level can accurately eva-luate the peroxidation damage in neonates with jaundice.
2.COX-2 induced angiogenesis and peritoneal adhesion
International Journal of Surgery 2008;35(8):524-528
Postoperative peritoneal adhesion represents a major complication of surgery. Recently, the angiogenesis which cyclooxygenase-2 enzyme induced was found to play an important role in the adhesion synthesis. This review summarized the relationship between COX-2 induced angiogenesis and peritoneal ad- hesion.
3.The correlation between chronic gastritis with benign mucosal nodular-change,Helicobacter pylori infection and lymphoid follicles infiltration
Yan KANG ; Xin CHEN ; Shi-Ying LI ;
Chinese Journal of Digestion 1998;0(06):-
Objective To study the correlation between chronic gastritis with benign mucosal nodular-change,Helicobacter pylori(H.pylori)infection and lymphoid follicles infiltration.Methods During July 1,2004 to June 30,2005 patients with chronic gastritis and benign mucosal nodular-change were identified by chromo-endoscopy with anabrosis indigo carmine staining at the antrum.Multiple biopsies were obtained for H.pylori detection with quick test and for pathology examination of mucosal lymphoid follicles formation and lymphocyte infiltration,as well as H.pylori infection.Results The patients were divided into nodular gastritis group,atrophy gastritis group and verrucous gastritis group with mean age of(31.00?11.62),(58.61?12.14)and(51.29?12.99)years old,respectively.The patients with nodular gastritis were the youngest(P
4.Path analysis of self-concept and depression on self-care in patients with chronic heart failure
Xiaofeng KANG ; Qingyin LI ; Xin SHAO ; Zheng LI
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(3):250-252
Objective To explore the role of depression as a moderator of self-concept on self-care behaviors in patients with chronic heart failure(CHF).Methods A sample of 160 in-patients with CHF were recruited from a cardiovascular hospital in Beijing.Data were collected by self-report questionnaires assessing self-care behaviors,self-concept and depression.Analysis of Moment Structures (AMOS) version 17.0 software was used to conduct the path analyses.Results Body sensation,self-consistency,self-ideal,moral-ethical-spiritual self and depression revealed direct effects on self-care behavior.Body sensation,self-ideal,moral-ethical-spiritual self affected self-care indirectly via depression.These paths' total coefficients were 0.09,0.15,0.31,0.09 and -0.26,respectively.Conclusion Self-care behaviors are directly affected by self-concept and depression.Depression has a greater moderating effect on these relationships.The findings provide a theoretical basis to direct the development of interventions for improving self-care in individuals with heart failure.
5.Construction and screening of human AQP1 shRNA expression vectors
Zhuo LI ; Wei KANG ; Na XIN ; Yu TIAN ; Jianhua LI
Chongqing Medicine 2015;(30):4183-4186
Objective To construct and screen effective shRNA expression vectors targeting human AQP1 gene ,and evaluate the interference efficiency of the AQP1 shRNA recombinant plasmids ,thus provide basis for further exploration on the effect and mechanism of AQP1 gene on human breast cancer cells .Methods Four pairs of shRNA sequences targeting human AQP1 gene were designed and synthesized ,and then inserted into the GV115 vector .AQP1 shRNA and control shRNA plasmids were trans‐fected into human breast cancer MCF‐7 cells .The expression of AQP1 mRNA and protein were detected by real time PCR(RT‐PCR) and Western blot to evaluate the interfering efficiency .Results RT‐PCR demonstrated that AQP1 was expressed in human breast cancer MCF‐7 cells .Sequencing showed that the shRNA vectors targeting AQP1 were successfully constructed .48 h after the AQP1 shRNA transfection ,AQP1 mRNA and protein expression levels in MCF‐7 cells were reduced to a significant degree ,and the AQP1 shRNA 4 plasmid vector could inhibit the AQP1 most efficiently .Conclusion The AQP1 shRNA recombinant plasmids vectors were successfully constructed and can significantly inhibit the expression of AQP1 in MCF‐7 human breast cancer cells .
6.Efficacy assessment of drug-loaded amniotic membrane transplantation for infectious corneal ulceration
Jing, LI ; Bing, LI ; Tong-xin, WEI ; Yu-guo, KANG
Chinese Journal of Experimental Ophthalmology 2013;(5):434-439
Background Treatment of corneal ulceration by transplanting drug-loaded amniotic membrane has been used widespreadly abroad,however,seldom study is found in China up to now.Objective This study was to explore the sustained release property and the efficacy of the drug-loaded amniotic membrane.Methods The bacteriostatic area of amniotic membrane fragments immersed with different concentrations (5,20,30 g/L) of levofloxacin for different time points (5,15,30,60 minutes) was evaluated by in vitro test.Bacterial corneal ulceration models were established in 20 rabbits by injected 0.7 MCF staphylococcus aureus suspension (0.1 ml) into the central corneal stroma to form the cloudy area of 4.0-6.0 mm.Then the rabbits were randomized into two groups.Regular amniotic membrane transplantation was performed laterally and 0.5% levofloxacin drops was topically administered after operation in the amnion+levofloxacin drops group,and drug-loading amnion transplantation was used in the drug-loading amnion group.Aqueous humor of 0.1 ml was collected in 30 minutes,1 hour,2,3.5,5.5 hours after levofloxacin was administered and 1 day,3,7,10,14,21 days after operation for the detect of levofloxacin level with high-performance liquid chromatography.The corneal symptom was scored based on McNeill's criteria in 1 week,2 weeks and 4 weeks and the ulceration area was assessed under the slit lamp in the first week.The pathological examination was carried out in the fourth week after surgery.Results The mean bacteriostatic area was bigger with the increase of levofloxacin concentration,and bacteriostatic area in amnion immersed for 15 minutes was bigger than that of 5 minutes (P<0.01).The levofloxacin concentration of aqueous humor after transplantation was decreased by extending the time,and that in 30 minutes and 5.5 hours after operation was (0.873±0.264) mg/L and (0.106±0.027) mg/L,respectively,in the amnion+levofloxacin drops group,and that in day 1,3,7 after surgery in the drug-loading amnion group was higher than at 30 minutes in the amnion+levofloxacin drops group,showing all significant differences (all P =0.00).In the first,second and fourth week after operation,the corneal symptom score was 1.7±0.6,1.3±0.5,0.2±0.4 in the drug-loading amnion group and 2.2±0.8,2.0±0.6,1.5±0.8 in the amnion+ levofloxacin drops group,with the significant differences among the different groups and time points (Fgroup =9.49,P =0.01 ;Ftime =22.96,P=0.01).The corneal ulceration area was (1.6±0.6) mm2 in the drug-loading amnion group and (3.2±0.8) mm2 in the amnion+levofloxacin drops group 1 week after operation,showing a significant difference between them (t =3.98,P =0.00).Histopathological revealed that the various layers of cornea tissue appeared irregular arrangement in the amnion + levofloxacin drops group 4 weeks after operation with 1-2 layers of new squamous epithelium.Disorder hypothallus structure,more inflammatory cells and residual vascular cavity were visible.However,new squamous epithelium of 4-5 layers was seen in the drug-loading amnion group,and inflammatory cells and residual vascular cavity were less than the amnion+levofloxacin drops group 4 weeks after operation.Conclusions Levofloxacin-loaded amniotic membrane can sustained release levofloxacin and maintain an effective drug concentration in aqueous humor,which improves the treating efficacy for staphylococcus aureus corneal ulceration.
7.Isolation and cultivation of goat temporomandibular joint disc cells
Weina SHU ; Hong KANG ; Weiping ZHANG ; Xin LI ; Guangjie BAO
Journal of Practical Stomatology 2010;26(2):165-168
Objective:To study the isolation, culture and identification of the TMJ cells and to observe the biological characteristics of cultured fibrochondrocytes. Methods:The TMJ discs were dissected from two 1 month goats under sterile conditions and were digested with collagenase. The cells were collected. Morphological changes and attachment efficiency were constantly observed under phase-contrast microscope. Immunohistochemical staining for type I collagen as well as toluidine blue staining were performed. Ultrastructures of the TMJ cells were observed under transmission electron microscope. Results: Most of the primary fibrochondrocytes presented a short spindle-shape while the rest showed polygon-shape. On the 7th day, the perliferating fibrochondrocytes started to contact each other to form a monolayer covering the bottom of the incubation disc. Immunohistochemical staining of type I and toluidine blue staining exhibited positive results. The fibrochondrocytes cytoplasms were rich in mictochondria and endoplasm reticulum. Conclusion: The fibrochondrcytes isolated from one-month-old goat TMJ disc have good proliferation ability in vitro and cells from passage 1 to 3 might be used as seed cells for TMJ disc tissue engineering.
8.Selective COX-2 inhibitor Celecoxib prevents postoperative abdominal adhesion
Kang WANG ; Xuqi LI ; Guangbing WEI ; Xin XU ; Gaofeng PING
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(2):217-220
Objective To determine the preventive effect of Celecoxib on postoperative adhesion formation and its mechanism. Methods We divided 80 SD rats into 5 groups: Groups A, B, C, H and S. Rats in Groups A, B, C and H received the operation of peritoneum rubbing to promote adhesion formation. Group S underwent sham operation. Rats in Group B were given Celecoxib of 40 mg/(kg·d), those in Group C were also given Celecoxib of 20mg/(kg·d), and those in Group H were given sodium hyaluronate (HA) during the operation. On the 8th and 15th postoperative day, half of the rats were sacrificed, the extent of adhesion formation was assessed and the adhesive peritoneum was subjected to immunohistochemistry with VEGF and CD_(34). Results The extent of postoperative adhesion differed significantly among the five groups (P<0.01). Groups B and C had significantly fewer adhesions than Groups H and A. VEGF was expressed most highly in Group A, followed by Groups H, C and B, and most weakly in Group S. CD34 was expressed most highly in Group A, followed by Groups H, C and B, and most weakly in Group S. Conclusion Celecoxib provides durable inhibition of intra-abdominal adhesions in a murine model compared with HA. The mechanism of preventing intra-abdominal adhesion via inhibiting COX-2 is possibly through down-regulated expression of VEGF and reduced microvascular density.
9.Establishment of leukocyte-associated immunoglobulin like-receptor 2 (CD306) eukaryotic expression vectors and purification and identification of fusion protein
Chunyan WANG ; Zhenhua KANG ; Xin XIE ; Yan LI ; Boquan JIN
Chinese Journal of Tissue Engineering Research 2009;13(50):9928-9932
BACKGROUND: Biological function of leukocyte-associated immunoglobulin like-receptor 1 (LAIR 1) has clearly researched in China and abroad, but the in vivo biological function of LAIR is poorly understood. OBJECTIVE: To establish LAIR-2 (CD306) eukaryotic expression vectors and to purify and identify the fusion protein. DESIGN, TIME AND SETTING: A single sample observation experiment was performed at the Fourth Military Medical University of Chinese PLA between June 2007 and June 2008.MATERIALS: plg/3c vector was offered by Oxford University, pcDNA3.1 vector was provided by Meyaard doctor. Chinese hamster ovary (CHO) cell lines were preserved by the Department of Immunology, Fourth Military Medical University of Chinese PLA.METHODS: Two eukaryotic expression vectors plg/3c-LAIR-2 and pcDNA3.1-LAIR-2 were constructed and were transfected into CHO cells. The binding activities of LAIR-2 fusion protein to LAIR-2 mAbs were identified by Western blot, immunocytochemistry and flow cytometry assay.MAIN OUTCOME MEASURES: The construction of stably transfected cell lines, and the purification and identification of fusion protein. The activity of LAIR 2 protein combined to corresponding monoclonal antibody.RESULTS: Eukaryotic expression vectors were constructed and trasnsfected into CHO cells successfully. Two cells lines CHO/LAIR-2-Fc and CHO/LAIR-2 that steadily expressed LAIR-2-Fc fusion protein and LAIR-2 protein were established. Western blot assay showed that LAIR-2 protein could bind specially to LAIR-2 mAb 1A7, 3H12 and 4A9. Immunocytochemistry and flow cytometry assay demonstrated that 3H12 and 4A9 could bind to LAIR-2 expressed in the transfected CHO cells. CONCLUSION: Two ceils lines CHO/LAIR-2-Fc and CHO/LAIR-2 were successfully constructed, which can transfected to CHO cells. The eukaryotic expressed LAIR-2 protein has good binding activity to LAIR-2 mAbs.
10.Type and distribution of cells in goat temporomandibular joint discs and its significance in tissue engineering design
Weina SHU ; Hong KANG ; Weiping ZHANG ; Xin LI ; Guangjie BAO
Chinese Journal of Tissue Engineering Research 2009;13(46):9022-9026
BACKGROUND:There is no common cognition in the cell type in the temporomandibular joint(TMJ)discs,and names describing TMJ disc cells also vary a lot.OBJECTIVE:To characterize the type and the distribution of cells in the TMJ disc of goats DESIGN,TIME AND SETTING:A single sample observation was completed in Cettutar and Motecutar Biologicat Center and Electron Microscope Center of Lanzhou University from March to May in 2007.MATERlALS:TMJ discs were obtained from two one-month-old healthy goats that were slaughtered freshly.METHODS:Bilateral TMJ discs of goats were cut off completely and were divided into 6 parts by 3 cuts in the major axis direction (mediolaterally)and 2 cuts in the minor axis direction(anteroposteriorly).Then the marked samples were fixed in 10%neutral formalin Iiquid for 24 hours and embedded by paraffin.MAIN OUTCOME MEASURES:Hematoxylin and eosin staining were used to identify regional variation of cell type and cellnumbers.Toluidine blue staining and collagen type Ⅰimmunohistochemical assay were performed to test the distribution of collagens.Transmission etectren microscopy was used to observe the ultrastructure of cells of goat TMJ discs.RESULTS:TMJ discs were comprised of cells and collagen fibers distributing unevenly.Collagens were mostly type Ⅰ.Collagen fibers were wave or crimping and approximately parallel to each other.with cells scattered in their matrix.Fibroblast-like cells and chondrocyte-like cells were the main two types of cells existing,with the former predominating over the later in a ratio of 2.05:1 approximately.There were no significant regional differences in cell type and distribution statistically.Transmission electron microscopy denoted that fibroblast-iike cells have fairly larger fusiform or irregular nuclei with very few organelles,while the chondrocyte-like cells exhibited round or elliptical nuclei,well defined pericellular electron lucent zones,unconspicuous cytocysta and non-distinctive pseudopodia CONCLUSION:There are no significant differences in type,number and arrangement of cells in TMJ discs of one-month-old goats statistically,with Fibroblast-like cells predominating slightly over chondrocyte-like cells.