1.Effect of clemastine fumarate on lung ischemia-reperfusion injury in rabbits
Wei WANG ; Ning LIU ; Huirong HAN ; Ning LI ; Rui ZHANG
Chinese Journal of Anesthesiology 2016;36(10):1193-1196
Objective To investigate the effect of clemastine fumarate on lung ischemia-reperfusion (I/R) injury in rabbits.Methods Fifty New Zealand white rabbits of both sexes,weighing 2.0-3.0 kg,were divided into 3 groups using a random number table:sham operation group (Sham group,n =10),I/R group (n =20) and clemastine fumarate group (Cle group,n =20).The model of lung I/R was established by clamping the left hilum of lung and decreasing the tidal volume followed by restoration of perfusion and ventilation 1 h later in I/R and Cle groups.At 3 h of ventilation in group Sham and 2 and 4 h of reperfusion in I/R and Cle groups,blood samples were collected for determination of serum tumor necrosis factoralpha (TNF-α) and interleukin-8 (IL-8) concentrations by enzyme-linked immunosorbent assay.The left lung was lavaged,and the broncho-alveolar lavage fluid (BALF) was colleted for determination of white blood cell count.Lung specimens were obtained for microscopic examination of the ultrastructure of lung tissues and for determination of wet/dry weight ratio (W/D ratio),expression of IL-1β and IL-6 mRNA (by real-time polymerase chain reaction) and cell apoptosis (by TUNEL).The apoptosis rate was calculated.Results Compared with Sham group,the W/D ratio,white blood cell count in BALF,serum concentrations of TNF-α and IL-8 and apoptosis rate were significantly increased,and the expression of IL-1β and IL-6 mRNA was up-regulated in I/R and Cle groups (P<0.05 or 0.01).Compared with I/R group,the W/D ratio,white blood cell count in BALF,serum concentrations of TNF-α and IL-8 and apoptosis rate were significantly decreased,and the expression of IL-1β and IL-6 mRNA was down-regulated in Cle group (P<0.05 or 0.01).The pathological changes of lung tissues were significantly attenuated in Cle group than in I/R group.Conclusion Clemastine fumarate can attenuate lung I/R injury in rabbits.
2.Bionic biphasic ceramic bioactive bone for segmental bone defects
Ning DUAN ; Wentao ZHANG ; Huiguang CHENG ; Ning JIAO ; Shaofeng HAN
Chinese Journal of Tissue Engineering Research 2015;(25):3952-3956
BACKGROUND:The bioceramics has the ideal pore size, high porosity and the through-hole rate, can provide the ideal physiological activity space for the bone cel repair, and can obviously improve bone conduction. OBJECTIVE:To explore the bone conduction and bone induction in the repair of bone defects in the stage of bone defect of bionic biphasic ceramic bioactive bone. METHODS:A total of 20 New Zealand white rabbits were randomly divided into bioactive glass and biomimetic biphasic ceramic bioactive bone groups, and were used to construct the animal bone damage model. They were given the repair with bioactive glass and biomimetic biphasic ceramic bioactive bone. RESULTS AND CONCLUSION:At 4 weeks after model establishment, scanning electron microscopy demonstrated that dense periosteal tissue was observed in the biomimetic biphasic ceramic bioactive bone group. At 8 weeks, dense combination was found, and no obvious fissure existed. At 12 weeks, complete bone demarcation blurred, showing a natural transition. Moreover, the binding site was very dense. There were a large number of new bone tissues, bone trabecula was regular and connected to a piece. The bone material has been largely degraded. Bone defects were repaired completely. The bone density was close to normal bone. At 8 weeks, in the bioactive glass group, the binding site presented obvious fissure. At 12 weeks, the fissure had been connected, but the binding was not tight as compared with the bionic biphasic ceramic biologic active bone group. The bone defect got preliminary repair. A smal number of new bone formed trabecular bone, but could not connect or traverse. There was no recanalization of the marrow cavity. A few continuous bone cal us traversed the broken end. These data demonstrate that bionic biphasic ceramic bioactive bone has good bone conduction, bone induction and biocompatibility in the repair of segmental bone defects.
3.Construction of two retroviral vectors for interleukin-1 receptor antagonist and interleukin-10 and in vitro transfection of rabbit synoviocytes
Fei HAN ; Ning ZHANG ; Fulin TANG
Chinese Journal of Rheumatology 2001;0(02):-
Objectives To construct two retroviral vectors, one containing human interleukin-1 recep-tor antagonist (hIL-1Ra) gene and the other containing human interleukin-10 (hIL-10) gene and to transfect rabbit synoviocytes in vitro and detect the expression level of target genes. Methods RNA from human peripheral blood mononuclear cells were extracted and target genes were amplified by RT-PCR. The target genes were cloned into retroviral vector pLXSN, which was then transducted into GP2-293 cells to produce recombinant retrovirus. Rabbit synoviocytes were transfected and the expression of target genes was detected by RT-PCR, immunohistochemistry and western-blot. Results The retroviral vector containing hIL-1Ra gene or hIL-10 gene was constructed successfully. The hIL-1Ra gene and hIL-10 gene were transduced respectively into rabbit synoviocytes in vitro. The mRNA level of both genes reached peak in 5 days. In positive cell clones, the protein level of hIL-1Ra reached peak within 30 days and maintained at least 60 days; the protein level of hIL-10 maintained at least 40 days. Conclusion The hIL-1Ra gene and hIL-10 gene can be transduced successfully into rabbit synoviocytes by recombinant retrovirus.
4.CKLF1 induces SH-SY5 Y cell migration via PLCγ/FAK signaling pathway
Zhenzhen WANG ; Yuhe YUAN ; Ning HAN ; Yi ZHANG ; Naihong CHEN
Chinese Pharmacological Bulletin 2014;(9):1209-1213
Aim To investigate the role of chemokine-like factor 1 ( CKLF1 ) in SH-SY5 Y cell migration and its molecular regulatory mechanism. Methods SH-SY5Y cells were stimulated with CKLF1 for 0. 5 h, 2 h, 8 h and 24 h, respectively. The migration distance and the percentage of migration cells were recorded by CELLocate analysis. The phosphorylation of focal ad-hesion kinase ( FAK) at Tyr-397 site was detected by Western blot analysis. By chemotaxis assays, we con-firmed the chemotaxis of CKLF1. Furthermore, FAK inhibitor PF-573228 and PLCγ inhibitor U73122 were used for the research of molecular regulatory mecha-nisms involved. Results CKLF1 promoted cell migra-tion and induced a strong increase in the phosphoryla-tion level of FAK-pY397 , which were significantly at-tenuated by the presence of U73122 ( a specific inhibi-tor for PLCγ) . In addition, the chemotaxis of CKLF1 was obviously blocked by the FAK inhibitor PF-573228 . Conclusion CKLF1 induces SH-SY5 Y cell migration via PLCγ/FAK signaling pathway.
7.Inflammatory myofibroblastic tumors of the urinary tract: a clinical diagnosis and treatment review in pediatric
Wenwen HAN ; Hongcheng SONG ; Weiping ZHANG ; Ning SUN
Chinese Journal of Urology 2016;37(4):292-295
Objective To improve the diagnosis and treatment of inflammatory myofibroblastic tumor (IMT) of the urinary tract in pediatric.Methods The retrospective study of 12 IMT was based on information retrieved from Beijing Children's Hospital from January 2006 to July 2015.The literatures of urinary IMT were reviewed.There were 12 cases of urinary IMT, with 8 cases in bladder, 2 in kidney, 1 in ureter and 1 in prostate.Mean age at surgery was 6.4 years old (range 2months-13 years), 6 cases males and 6 females.Tumor resection were performed in 11 patients, biopsy was performed only in 1 patient.Results HE staining revealed diffuse appearing spindle myofibroblastic cells admixed with inflammatory cells.Immunohistochemistry showed positive ration for following markers as ALK (8/12), CK18 (6/12), Desmin (7/12), SMA (8/12), Actin (1/2), Vimentin (9/12).Negative staining were seen for Myoglobin, S-100 and Ki-67 < 20%.Patients were followed up in 10 cases, lost to follow-up in 2;the mean follow-up time was 14.4 months (range 3-31 months).All patients recovered well without relapse or metastasis.Condusions Inflammatory myofibroblastic tumors of the urinary tract in pediatric were rare, without specific characteristic in clinical features and imaging.The main treatment of IMT is complete surgical excision.
8.Effects of Clemastine Fumarate on Expression of Toll Like Receptor 4 in Lung Ischemia-Reperfusion
Ning LIU ; Wei WANG ; Rui ZHANG ; Huirong HAN
The Journal of Practical Medicine 2016;32(18):2988-2991
Objective To explore the expression change of Toll Like Receptor 4 (TLR4) in lung ischemia-reperfusion injury in rabbits and the influence of clemastine fumarate on it. Methods Fifty rabbits were divided into five groups randomly (n=10): Sham (N+S), reperfusion for 2 hours (N+I1/R2), reperfusion for 4 hours (N+I1/R4), clemastine fumarate + reperfusion for 2 hours (F+I1/R2) and clemastine fumarate+reperfusion for 4 hours (F+I1/R4). The ischemia time in each group was 1 hour and normal saline was given respectively in groups of N+S , N+I1/R2 and N+I1/R4. Western blotting , RT-PCR and immunofluorescence were used to detect the expression of TLR4 in lung tissue , and the changes of ultrastructure in ischemia-reperfusion lung tissue were observed by electron microscope. Result The expression of TLR4 was elevated obviously in ischemia-reperfusion lung tissue (P<0.05), and there was positive correlation between the increased TLR4 level and reperfusion time (P<0.05), the swelled and thick-ridge mitochondria were observed in type II alveolar epithelial cells after LIRI (P<0.05); but clemastine fumarate inhibited the expression of TLR4 in lung tissue significantly caused by LIRI (P<0.05). And the mitochondria injury was reduced in the groups of clemastine fumarate. Conclusion TLR4 expression is elevated in lung tissue after LIRI; clemastine fumarate inhibits the expression of TLR4 caused by LIRI and protects the lung tissue from LIRI in rabbits.
9.The experience of diagnosis and mangement in coexisting ureteropelvic junction obstruction and nonreflux megaureter
Wenwen HAN ; Hongcheng SONG ; Weiping ZHANG ; Ning SUN ; Chengru HUANG
Chinese Journal of Urology 2017;38(2):95-98
Objective To investigate the experience of diagnosis and management of coexisting ureteropelvic junction obstruction (UPJO) and nonreflux megaureter (NRM).Methods The retrospective study of UPJO with NRM was based on 10 years information retrieved from January 2005 to December 2015.The data of 13 patients (8 males and 5 females) were available and recorded.Mean age at surgery was 3.7 years old (range 1.8 to 14 years).The diagnosis and mangement were summarized.Coexisting ureterovesical junction obstruction (UVJO) and vesicoureteral reflux,iatrogenic stricture and vesicoureteral reflux were excluded.Intravenous pyelography,voiding cystourethrography,ultrasound and CT reconstruction were performed before operation.Only six patients had an accurate diagnosis as UPJO with UVJO before surgery.Pyeloplasty was the initial surgical management choice for 10 patients,and ureteroneocystostomy in 3 patients.Results UVJO were diagnosed with pyelography techniques in 3 patients after pyeloplasty,while 4 were diagnosed as nonreflux and nonobstruction megaureter.Of the 10 patients who underwent initial pyeloplasty,additional ureteroneocystostomy was required in 3 and the prognosis was good.Additional pyeloplasty was required in 2 of the 3 patients who initially underwent ureteroneoeystostomy.Mean follow-up time from last operation was 23.3 months (6-53 months),the overall prognosis was good.Conclusions It is often difficult to correctly diagnose coexisting UPJO and NRM.In patients with UPJO,it is highly recommended nephrostomy radiography after pyeloplasty to evaluate the distal ureterovesical junction.Initial pyeloplasty is always recommended as first-line therapy.Additional ureteroneocystostomy was required when hydroureteropelvic was aggravated.
10.Primary malignant rhabdoid tumor of the ureter: a case report and literature review
Wenwen HAN ; Weiping ZHANG ; Ning SUN ; Hongcheng SONG ; Chengru HUANG
Chinese Journal of Urology 2015;36(11):818-821
Objective To discuss the clinical and pathological features of malignant rhabdoid tumor of the ureter (MRTU).Methods One case of MRTU was reported, a six-year-old girl was admitted to our hospital on May 29, 2014, and presented left loin pain 2 weeks, ultrasound showed gradually progressing hydronephrosis and hydroureter.During a physical examination, she felt tenderness in the left kidney area and no mass was palpable in abdomen.The ultrasound showed left sided gross hydroureteronephrosis and a round hyperechogenic mass in the inferior pole of the left ureter (In front of the left iliac vessel), with no obvious borders.Contrast-enhanced CT suggested a gross dilatation of the left kidney and ureter with a solidappearing lesion in the lower ureter;neither additional abdominal abnormalities nor enlarged lymph nodes were seen in both examinations.The surgery began with incision of left lower abdomen.The partial ureter of neoplasm was excised along with invaded psoas and posterior peritoneum by gross inspection, then ureteroureterostomy was performed.The severed ureter was completely blocked with the ill-defined neoplasm and was 3.3 cm in length and 2.1 cm in width.Results The ureteral neoplasm was excised,along with the invaded psoas and posterior peritoneum,after that ureteroureterostomy was performed.HE showed the diffuse large round nuclei, vesicular chromatin, prominent nucleoli cells, and moderate amounts of eccentrically placed eosinophilic cytoplasm.Immunohistochemical studies were positive for cytokeratin, epithelial membrance antigen and vimentin, negative for INI1, METU hereby was confirmed.She underwent a chemotherapy regimen consisting of ICE, alternating with VDC.Four courses chemotherapy (3 months) later,CT scan suggested hematogenous metastasis of lung.The family refused further treatment and the patient died of systemic metastasis eight months after surgery.Conclusion MRTU was a rare and highly aggressive tumor with a poor prognosis.