2.Up-regulation of Notch1 inhibits proliferation and differentiation of osteoclast in vitro.
Yilin PING ; Feng LOU ; Xiao YANG ; Ping ZHANG
West China Journal of Stomatology 2016;34(2):121-124
OBJECTIVEThis study aimed to explore the effect of the up-regulation of Notch1 on osteoclastogenesis induced to osteoclasts by receptor activator for nuclear factor-kappaB ligand (RANKL) and macrophage colony-stimulating factors (MCSF) in vitro.
METHODSThe bone marrow stem cells (BMSCs) of Rosa(-notch1) mice were cultured and induced to osteoclasts by RANKL and MCSF. The BMSCs were transfected with the Ad-Cre-green fluorescent protein (GFP) virus or Ad-GFP virus. Total RNA from cells was extracted, and the gene expression levels of Notch1, Notch2, Notch3, Notch4, Deltal, Delta3, Delta4, Jagged1, Hes1, and tartrate resistant acid phosphatase (TRAP) were detected at the defined stage by reverse transcription-polymerase chain reaction (RT-PCR). Osteoclast formation was analyzed by TRAP assay.
RESULTSThe number of TRAP-positive multinuclear cells of the experimental group significantly decreased compared with that of the control group. The mRNA expression levels of Notch1, Notch3, Jagged1, Delta3, and Hesl of the experimental group were significantly higher than those of the control group, whereas the TRAP mRNA expression of the experimental group was significantly lower than that of the control group (P<0.05).
CONCLUSIONUp-regulation of Notch1 inhibit osteoclastogenesis of BMSCs induced by RANKL and MCSF in vitro.
Animals ; Cell Differentiation ; Cell Line ; In Vitro Techniques ; Macrophage Colony-Stimulating Factor ; Mice ; Osteoclasts ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Receptor, Notch1 ; metabolism ; Receptor, Notch2 ; Up-Regulation ; physiology
3.Application of emergency transcatheter arterial embolization on liver and spleen rupture
Jianke YANG ; Ping JIN ; Weiyin LOU ; Defeng ZHANG ; Jiantin ZHU
Chinese Journal of Primary Medicine and Pharmacy 2014;(22):3406-3407
Objective To investigate the value of emergency transcatheter arterial embolization in the treat-ment of liver and spleen rupture,in order to provide the reference for clinical therapeutic strategies.Methods In our hospital,58 patients with liver and spleen rupture were selected.Operation time,catheter angiography,embolization of success rate,postoperative survival rates and the postoperative complications were recorded.Then follow-up,patients with sequela of occurrence was recorded.Results Embolizations of 55 cases were successful,3 cases accounted for 94.8%,2 were successful, no embolization failed, the success rate of catheter angiography in the diagnosis was 100.0%.31 cases of arterial embolization for splenic artery,10 cases of hepatic artery,8 cases of right hepatic artery, 2 cases of left hepatic artery.The average operation time was (57.8 ±15.6) min.All the patients were successfully hemostatic,postoperative survival rate was 100.0%for patients.2 cases of postoperative biliary leakage,1 cases with splenic abscess,the incidence of postoperative complications was 5.2%.Recovery of all patients was good,without the occurrence of sequelae.Conclusion Rupture has good clinical efficacy of emergency transcatheter arterial emboliza-tion in treatment of liver and spleen,liver,spleen can be retained in the circumstances,the effective control of hemor-rhage,improve the clinical success rate.
4.Establishment of an in vitro model of brain blood barrier
Yunan YANG ; Yunhua GAO ; Jinning LOU ; Ping LIU ; Chixue FU
Journal of Third Military Medical University 2003;0(22):-
Objective To study the dynamic changes of transendothelial electrical resistance (TEER) on our in vitro model of brain blood barrier (BBB) made from primary culture of BALB/c mouse brain microvascular endothelial cells (BMVEC), and to explore the relation between TEER and BBB permeability, and search for the best culture condition. Methods BMVEC were isolated from BALB/c mouse and cultured on a transwell insert with special micro-pore. The cells were identified with immunohistochemical methods and electron microscope. TEER over BMVEC was measured after BBB model establishment for determining the 3H-Glucose permeability of BBB in vitro. Results BMVEC cultured in the transwell insert exhibited typical "flagstone" appearance and in a tight monolayer structure under electron microscope. Immunohistochemical detection of ZO-1 protein, a marker antigen of tight junction, showed smooth, continuous and tight junctions between confluent BMVEC. TEER over BMVEC monolayer increased to (346?10) ?/cm2 when the permeability for 3H-Glucose was decreased to the minimum. Conclusion BBB model in vitro made from primary culture of BMVEC in transwell has the basic characteristics of BBB in morphology, electrical resistance and permeability.
5.Analysis of pathogens distribution and drug resistance from blood culture in a grade 3 A hospital
Xiaoyan LOU ; Weifeng JIN ; Lili ZHU ; Shuzi CHEN ; Qinglan YANG ; Ping LI ; Ping LIN
International Journal of Laboratory Medicine 2016;37(23):3263-3265,3268
Objective To analyze the distribution and drug resistance of pathogens isolated from blood culture specimens in a grade 3A hospital so as to provide a basis for the prevention and control of bloodstream infection ,and rational antibacterial drugs use .Methods A total of 3 241 blood culture results in our center from June 2014 to September 2016 were analyzed retrospectively . The blood samples were cultivated with the BacT/ALERT 3D and VersaTrek instruments .The bacterial identification and antibiotic susceptibility tests were conducted with VITEK‐2 Compact and ARIS 2X instruments .Results Among 3 241 blood culture speci‐mens ,99 strains of pathogenic bateria were isolated with the positive rate of 3 .1% ,including 42(42 .4% ) strains of Gram‐negative bacteria ,54(54 .5% ) strains of Gram‐positive bacteria and 3(3 .0% ) strains of fungus .The top three of Gram‐negative bacteria were in turn Eescherichia coli ,Klebsiella pneumonia and Pseudomonas aeruginosa ;the top three of Gram‐positive bacteria were in turn coagulase‐negative staphylococcus (CNS ) ,Staphylococcus aureus and streptococcus pyogenes .Enterobacteriaceae remained 100 .0% sensitivity to imipenem ,meropenem ,piperacillin/tazobactam and amikacin .The resistance rate of Gram‐positive bacterium to vancomycin ,linezolid ,teicoplanin and gentamycin was 0 .0% ,the rest had different degrees of drug resistance .Conclusion Ee‐scherichia coli ,CNS and Staphylococcus aureus are the most frequent pathogenic bacteria of the blood stream infection in our hospi‐tal .Pathogens show different resistance to different kinds of antibacterial agents .Clinic should rationally select the antibacterial a‐gents according to the drug susceptibility test results so as to slow down the generation of drug resistance bacterial strains .
6.Mechanism on the role of caspase recruitment domain-containing protein 9 in the early stage of acute pancreatitis
Chengzhao WEN ; Ping XU ; Zhiwen YANG ; Jing WANG ; Xiaoxiao MENG ; Xiaoli LOU
Chinese Journal of Digestion 2015;(9):624-629
Objective To investigate the mechanism of caspase recruitment domain‐containing protein 9 (CARD9) in the early stage of acute pancreatitis(AP) .Methods Peripheral blood mononuclear cells (PBMC ) of 49 AP patients (33 mild acute pancreatitis (MAP ) patients and 16 severe acute pancreatitis (SAP) patients) were collected on the Day 1st ,3rd and 5th of hospitalization .Twenty healthy volunteers were enrolled in control group .The expression level of CARD9 ,B‐cell lymphoma(Bcl)‐10 ,p38 mitogen‐activated protein kinase (MAPK ) and p65 nuclear factor Kappa B (NF‐κB ) in PBMC of AP patients were detected by Western blotting .The co‐localization ,expression and binding between CARD9 and Bcl‐10 in PBMC of control group ,SAP group and MAP group on the Day 1st hospitalization were determined by cell immune‐fluorescence staining and co‐immuno precipitation method .Single factor analysis of variance and Mann‐Whitney test were performed for data comparison between groups .Pearson method was used for correlation analysis .Results The results of Western blotting indicated that the expression of CARD9 and Bcl‐10 in PBMC of SAP group on the Day 1st ,3rd and 5th of hospitalization (1 .12 ± 0 .05 ,1 .03 ± 0 .03 and 1 .01 ± 0 .01 ;1 .74 ± 0 .08 ,1 .72 ± 0 .10 and 1 .69 ± 0 .11) were all significantly higher than those of control group (0 .33 ± 0 .10 and 1 .02 ± 0 .11) and MAP group (0 .71 ± 0 .02 ,0 .55 ± 0 .06 and 0 .25 ± 0 .07 ;1 .15 ± 0 .03 ,1 .09 ± 0 .07 and 1 .01 ± 0 .04) ,and the differences were statistically significant (F= 35 .76 and 18 .20 ,all P< 0 .05) .The expression of p38 MAPK in PBMC of SAP group on the Day lst ,3rd of hospitalization (1 .88 ± 0 .08 、1 .68 ± 0 .11) were significantly higher than those of MAP group on the Day 1st ,3rd ,5th (0 .86 ± 0 .08 ,0 .77 ± 0 .10 ,0 .73 ± 0 .20) and healthy control group (0 .58 ± 0 .24 , F= 7 .24 ,all P < 0 .01) .The expression of p65 NF‐κB in PBMC of SAP group on the Day 1st ,3rd of hospitalization (1 .64 ± 0 .02 ,1 .55 ± 0 .03) were significantly higher than those of MAP group on the Day 3rd ,5th (1 .06 ± 0 .14 ,0 .87 ± 0 .20) and healthy control group (1 .17 ± 0 .13 ,F= 4 .51 ,all P< 0 .05) .The results of immune‐fluorescence staining indicated that CARD9 and Bcl‐10 co‐localized in nucleus .The results of co‐immuno precipitation showed that the binding degree between CARD9 and Bcl‐10 of SAP group was significantly higher than that of control group and MAP group . Pearson correlative analysis suggested that the level of p65 NF‐κB and p38 MAPK in PBMC of AP patients were positive correlated with the expression of CARD9 (r= 0 .692 and 0 .834 ,both P< 0 .01) .Conclusion CARD9 is positive correlated with NF‐κB and MAPK , which indicates CARD9 induced inflammatory cytokines by activating NF‐κB and MAPK signaling pathways in AP .
7.Clinicopathologic features of 234 cases with borderline ovarian tumors
Fie REN ; Jiangyan LOU ; Hui LIU ; Ping WANG ; Jiawen ZHANG ; Kaixuan YANG ; Hongjing WANG ; Mingrong QIE ; Zhilan PENG
Chinese Journal of Obstetrics and Gynecology 2009;44(2):116-120
Objective To determine the clinicopathologic characteristics and prognostic factors that may be used to predict the poor outcome of patients with borderline ovarian tumors. Methods All cases with borderline ovarian tumors treated in the West China Second University Hospital from January 2001 to June 2007 were analyzed retrospectively for elinicopathologic features, treatment parameters and outcome of treatment. Univariate and multivariate analyses were used to assess independent prognostic factors using the logistic regression model. Results The median age of 234 patients was 40. 1 years with a range of 14 to 80 years. There were 101 (43.2%), 94 (40.2% ) , 19 (8.1% ), 12 (5.1%) , 8 (3.4%) cases of serous, mutinous, mixed, endometrioid and clear cell tumors, respectively. Out of 234 cases, 182 (77.8%) underwent laparotomy and 45 ( 19.2% ) underwent laparoscopy. Seven women underwent laparoconversion. Fertility sparing surgery was performed on 119 cases (50.9% ) and radical surgery was performed on 115 cases (49.1% ). Totally 161 (68.8% ) patients had stage Ⅰ , 19 ( 8.1% ) had stage Ⅱ, 54 ( 23.1% ) had stage Ⅲ, and none had stage Ⅳ disease. Sixty-four women received postoperative chemotherapy. The median follow-up was 40 months with a range of 8 to 78 months. Recurrence was found in 26 cases (11.1%) during follow-up, and no tumor-related death was reported. The logistic regression model showed that surgery procedure ( OR=2.304, P=0.024), cyst rupture ( OR=2.213, P=0.038 ), stage ( OR= 4.114, P<0.01 ), microinvasion ( OR=2.291, P=0.046) and peritoneal implants ( OR=2.101, P = 0.016) were the five independent prognostic factors affecting recurrence. Conclusions Although patients with borderline ovarian tumors have an excellent prognosis, the risk of recurrence remains in some patients. Emphasis should be put on these patients with high risk factors and preventive strategies should be taken to prevent their progression.
8.Influence of an anti-photosensitivity mixture on acute photodamage-induced cell apoptosis in BALB/c
Liyun SUN ; Ping WANG ; Bohua YANG ; Guangzhong ZHANG ; Jing WANG ; Shuping LI ; Kuiying LIU ; Weihai LOU ; Weiwen CHEN
Chinese Journal of Dermatology 2011;44(2):138-140
Objective To investigate the action mechanism of an anti-photosensitivity mixture on skin photodamage. Methods Twenty-eight BLAB/c mice were divided into 4 groups, i.e., normal control group,treatment group, negative and positive control groups; the last three groups were irradiated with a single dose of UVB at 300 mJ/cm2 after 7-day pretreatment with sodium chloride physiological solution, anti-photosensitivity mixture, and hydroxychloroquine, respectively. Twenty-four hours after the irradiation, mice were killed and skin tissue samples were obtained at the irradiated sites. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) and immunohistochemical staining were carried out to detect cell apoptosis,Fas and Caspase-3 protein expressions respectively. Results An increase was observed in the expression level of Fas and Caspase-3 and in the apoptotic index in keratinocytes from UV-irradiated mice compared with unirradiated control mice (all P < 0.01 ). In comparison with sodium chloride physiological solution, the antiphotosensitivity mixture suppressed the UV irradiation-induced increase in the expression intensity of Fas and Caspase-3 and apoptotic index in keratinocytes (P < 0.05 or 0.01 ). Conclusions The anti-photosensitivity mixture could alleviate UV-induced inflammatory damage to and apoptosis in epidermal keratinocytes, likely by regulating cell apoptosis and Caspase-3 pathway.
9.Anti-tumor activity of Actinidia chinensis planch
Li-Jun LOU ; Ding-Liang LV ; Zeng-Ren HU ; Song-Yang ZHONG ; Wei-Dong TANG ; Yong-Ping XU ;
Chinese Pharmacological Bulletin 1986;0(06):-
Aim To evaluate the anti-tumor activity of the extract from the root of Actinidia chinensis planch in vitro and in vivo. Methods Active components from the root of Actinidia chinensis planch were isolated by traditional phytochemical techniques. The in vitro anti-tumor activity was determined by sulforhodamine B assay and the in vivo anti-tumor activity was evaluated using experimental mouse tumor models and human tumor xenografts in nude mice. Results Powdered air-dried roots of Actinidia chinensis planch were percolated with methanol at room temperature thrice. The filtrate was concentrated to dryness in vacuo and then was further extracted with ethyl acetate, n-butanol , and chloroform. The fraction extracted by chloroform displayed the most potent activity against several tumor cell lines including hepatocellular carcinoma Bel-7402 cells, non-small cell lung cancer A549 cells, lymphoma Ramos cells, and breast cancer MCF-7 cells. Further more, the anti-tumor efficacy of the chloroform fraction was confirmed in Bel-7402 xenografts in nude mice with the percentage inhibition of 38.0 %. Conclusion The extract of the root of Actinidia chinensis planch has anti-tumor activity, and the active components are mainly in the fraction extracted by chloroform.
10.The sequence of fat deposit and its relationship with insulin resistance
Xiu-Ping BAI ; Hong-Liang LI ; Wen-Ying YANG ; Jian-Zhong XIAO ; Bing WANG ; Rui-Qin DU ; Da-Jun LOU ;
Chinese Journal of Endocrinology and Metabolism 1986;0(04):-
Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P