1.The expression of human CD55 and CD55-TM molecules in mouse fibroblasts cell line NIH3T3 and their function in complement lysis restriction
Man JIANG ; Ruiqin DU ; Wanling LI
Chinese Journal of Immunology 1999;0(12):-
Objective:To express the GPI-anchored CD55 and recombinant transmembrane form CD55 molecules on mouse fibroblasts cell line NIH3T3, and compare their inhibitory function of complement lysis.Methods:In previous study,had constructed the transmembrane form CD55 (CD55-TM) cDNA by linking the extracellular portion of CD55 to the transmembrane and cytoplasmic domains of MCP, and then subcloned into retroviral vector pLXSN. In this experiment,had transfected recombinant CD55-TM and GPI anchored CD55 into PA317 packaging cell to generate stable virus-producing cell lines. And then, mouse fibroblasts cell line NIH3T3 was infected with the virus containing CD55, recombinant CD55-TM or pLXSN alone. The expression of these molecules on NIH3T3 cells was detected by FACS analysis. Complement killing assay was carried out using MTT colorimetric method.Results:FACS analysis showed that both CD55 and its TM version were expressed stably on NIH3T3 cells. Both kinds of CD55 molecules can efficiently protect the NIH3T3 cells from complement mediated lysis and there is no significant difference between them. Conclusion:These data showed that both GPI-anchored CD55 and its TM version can normally expressed on NIH3T3 cells and both can protect the NIH3T3 cells from human complement mediated lysis. These results confirmed the feasibility of TM CD55 based gene therapy could be used for PNH, and also provided an excellent model for the study of signal transduction mechanisms mediated by GPI-anchored protein.
2.Role of glutamate receptor subtypes in cardiac nociception in nucleus tractus solitarius in rats
Xiaohua LIU ; Man HAN ; Jianqing DU
Journal of Jilin University(Medicine Edition) 2014;(3):508-512
Objective To study the role of glutamate receptor subtypes in nucleus tractus solitarius(NTS)in cardiac-somatic motor reflex (CMR)induced by intrapericardial administration of capsaicin,and to clarify the modulation mechanism of NTS to cardiac nociceptoion.Methods 60 SD rats were randomly divided into ibotenic (IBO)group, glutamate group, MK-801 group, MCGP group, MK-801 + MCPG group and DNQX group. The NTS microinjected with 130 mmol·L-1 IBO 100 nL,100,200,500 mmol·L-1 L-glutamate 100 nL,NMDA receptor antagonist 40 and 60 mmol · L-1 MK-801 100 nL, metabotropic glutamate receptors antagonist 25 and 50 mmol·L-1 MCPG 100 nL,25 mmol· L-1 MCPG 50 nL plus 40 mmol· L-1 MK-801 50 nL,non-NMDA receptor antagonist 20 and 50 mmol·L-1 DNQX 100 nL,respectively.The changes of CMR of the rats in various groups were observed.Results Compared with control group,the CMR of the rats in IBO group was decreased (P<0.05);the CMR was increased as the concentration increased in glutamate group(P<0.05);the CMR in MK-801 and MCPG groups were decreased (P<0.05);the CMR in MK-801+MCPG group was decreased (P<0.05);the CMR in DXQX group had no changes (P>0.05).Conclusion NTS play an facilictory role in cardiac nociception,and the NMDA receptors and mGluRs receptors mediate this facilitory modulation.
3.Regulation of metabotropic glutamate subtype 7 and 8 receptors in nucleus tractus solitarius in cardiac nociception in rats
Xiaohua LIU ; Man HAN ; Jianqing DU
Journal of Jilin University(Medicine Edition) 2014;(5):920-924
Objective To explore the role of metabotropic glutamate receptors (mGluRs)group Ⅲ and its subtypes mGluR7 and mGluR8 in nucleus tractus solitarius(NTS)in cardiac-somatic motor reflex (CMR),and to clarify the modulation role of mGluR Ⅲ and its subtypes in NTS in cardiac nociceptoion.Methods 40 SD rats were randomly divided into L-AP4 group,microinjection of mGluRs Ⅲ agonist L-AP4 0.1,1.0,10.0 or 20.0 nmol in NTS;AMN082 group,microinjection of mGluR7 agonist AMN082 1,2 or 4 nmol;DCPG group,microinjection of mGluR8 agonist DCPG 4, 6 or 8 nmol;MSOP group, microinjection of mGluR Ⅲ antagonist MSOP 20 or 100 nmol,20 nmol MSOP+410 nmol L-AP,20 nmol MSOP+2 nmol AMN082,20 nmol MSOP+6 nmol DCPG. The changes of CMR of the rats in various groups were observed.Results Compared with control,the CMR in L-AP4 and AMN082 groups was decreased (P<0.05);the CMR in DCPG group was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP had no change (P>0.05);the CMR in MSOP group after injection of 100 nmol MSOP was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP followed L-AP4 or AMNO82 had no change (P>0.05).Conclusion The group Ⅲ mGluRs in the NTS play an inhibitory role in cardiac nociception, and mGluR7 has anti-nociceptive effects while mGluR8 has pro-nociceptive effects.
4.Small cell carcinoma of the bladder: dagnosis and treatment of 18 cases
Du SHI ; Chuize KONG ; Zhenhua LI ; Wei ZHAO ; Xiaojun MAN
Chinese Journal of Urology 2016;37(8):607-610
Objective To study the clinical features,diagnosis and management of small cell carcinoma of the bladder (SCCB).Method The clinical data of 18 cases of patients with small cell carcinoma of the bladder were analyzed retrospectively and the literature were reviewed.There were 16 males and 2 females,ages 54 to 81 years (median age,61 years).Clinical manifestations included gross hematuria in 11 cases,urgency in 2 cases,dysuria in 2 cases and postoperative review after TURBT of bladder urothelial carcinoma in 3 cases.The median tumor size was 3.35cm (ranged,1.0 to 6.0 cm).2 cases underwent TURBT and intravesical chemotherapy regularly were followed after surgery.3 cases underwent partial cystectomy,intravenous chemotherapy combined with radiotherapy was followed in one case,the other 2 cases refused the following therapy.13 cases underwent radical cystectomy,intravenous chemotherapy was followed in 2 cases,pelvic radiotherapy was followed in 2 csaes and intravenous chemotherapy combined with radiotherapy was followed in 2 cases,the other 7 cases refused the following therapy.Results 11 cases were pure SCCB,7 cases were mixed SCCB,all with urothelial carcinoma.T1N0M0 in 3 cases,T2N0.1M0 in 4 cases,and T3N0-2M0 in 11 cases.The duration of follow-up was from 5 to 35 months after surgery.9 cases died of tumor metastasis,9 cases are still alive,except 1 case with lymph node metastasis,the other 7 cases are free of tumor recurrence or metastasis.Conclusions SCCB is rare,with high malignant degree and poor prognosis.The prognosis of the pure SCCB may be worse than the mixed SCCB.The diagnosis depends on pathology examination.Radical cystectomy is the main treatment method,the strategy of bladder-preserving may be an attempt for proper SCCB patients.Adjuvant therapy plus surgery may be better.
5.Management of mandibular symphysis fracture associated with superolateral dislocation of the mandibular condyle: 10 case report
Shuangquan DU ; Lian JIANG ; Xiaobo XIANG ; Cheng MAN
Journal of Practical Stomatology 2016;32(5):732-734
Mandibular symphysis fracture combined with anteromedial dislocated condyle fracture is commom in clinical,but mandibular symphysis fracture associated with superolateral dislocation of the mandibular condyle is rare,which is often misdiagnosed or completely over-looked.Malpractice can lead to ankylosis and other sequelae.This article reviews 10 patients with mandibular symphysis fracture associated with superolateral dislocation of the mandibular condyle,discusses the causative mechanism,diagnostic features and clinical management according to literature data.
6.Comparison of fluorescence staining and periodic acid-Schiff staining in the diagnosis of fungal keratitis
Man DU ; Li ZHANG ; Peng LI ; Hongmei QI ; Xiuhai LU
Chinese Journal of Experimental Ophthalmology 2021;39(2):119-125
Objective:To compare the effect of fluorescence staining and periodic acid-Schiff staining in the diagnosis of fungal keratitis (FK).Methods:A total of 147 corneal specimens from 147 FK patients treated in Eye Hospital of Shandong First Medical University from January 2017 to May 2019 with positive corneal scraping or fungal culture were collected.Among them, there were 84 cases with penetrating keratoplasty (PKP), 42 cases with lamellar keratoplasty (LKP) and 21 cases with lesion resection.Another 11 cases with herpes simplex virus keratitis served as negative control.The corneal tissue specimens were performed with fungal fluorescence staining and periodic acid-Schiff staining, respectively.The stained sections were placed under fluorescence microscope and optical microscope to observe fungal hyphae or spores, respectively.The positive rates of the two staining methods were compared, and the positive cases of the diagnosis of FK in corneal tissue samples obtained by different surgical methods and corneal infection caused by different strains of the two staining methods were compared.Written informed consent was obtained from each patient.The study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY-2016012).Results:The positive rate of periodic acid-Schiff staining and fungal fluorescence staining was 60.5% (89/147) and 79.6% (117/147), respectively.The positive rate of fluorescence staining in the diagnosis of fungal keratitis was significantly higher than that of periodic acid-Schiff staining ( χ2=28.00, P<0.01), and both the specificity of the two staining methods was 100%.The positive rate of specimens obtained by PKP with fluorescent staining was 85.7% (72/84), and the positive rate with periodic acid-Schiff staining was 65.5% (55/84), and the difference was statistically significant ( χ2=17.00, P<0.01). The positive rate of specimens obtained by LKP with fluorescent staining was 71.4% (30/42), and the positive rate with periodic acid-Schiff staining was 52.4% (22/42), and the difference was statistically significant ( χ2=8.00, P<0.01). The positive rate of resected foci specimens with fluorescent staining was 71.4% (15/21), and the positive rate with periodic acid-Schiff staining was 57.1% (12/21), and the difference was not statistically significant ( χ2=1.30, P=0.25). The positive cases of two kinds of staining were different among different fungal strains.Among them, the positive cases of Fusarium solani complex, Pythium insidiosum, Aspergillus fumigatus complex, Candida guilliermondii, Trichoderma and Nigrograna mackinnonii with fluorescence staining were 19, 5, 5, 1, 1 and 1, and the positive cases of periodic acid-Schiff staining were 11, 0, 3, 0, 0 and 0, respectively.The staining results of the 11 negative controls were negative. Conclusions:Fluorescence staining is more sensitive than periodic acid-Schiff staining in the detection of fungal components in paraffin-embedded corneal tissues, and it can significantly improve the fungal detection rates.
7.Investigation of the value of multi-slice CT perfusion imaging on brain hemodynamic in patients with cerebral infarction
Zhenhua LIU ; Yifeng DU ; Jingguang LV ; Lin LU ; Xiao MAN ; Jianping CHEN ; Zhenfang LIU
Chinese Journal of Geriatrics 2011;30(6):452-454
Objective To study the value of CT perfusion imaging(CTPI)on brain hemodynamic of the aged with cerebral infarction. Methods The 48 patients who were doubted with cerebral infarction underwent 16-slice CT plain scanning and CTPI within 24 hours of onset. The cerebral blood flow(CBF), mean transit time(MTT)and time to peak(TTP)of contrast-medium in region of interest(FOV)were used as brain hemodynamic parameters in comparation with contralateral regions. All cases were followed up with MRI after 3-10 days. Results Ischemia lesion was found on CT plain scanning in 40.9% of patients, while 93.2% of patients showed abnormal perfusion on CTPI. The sensibility of CTPI in identifying ischemia area was 93.2%, and the specificity was 100%. CBF in research area was significantly reduced, MTT and TTP were remarkably increased in contrast to counterparts(P<0.01). Conclusions CT perfusion imaging can sensitively reveal the hemodynamic condition of cerebral ischemia, which could provide the important information for early diagnosis and treatment of the elderly with brain infarction.
8.Effect of ERK1/2 on ischemic preconditioning in diabetic rat hearts
Caiyun MA ; Like ZHANG ; Fenghe DU ; Lingqiao LU ; Hongxia WANG ; Xiangjun ZENG ; Ping FU ; Qingqing MAN
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To test the effect of ERK1/2 on ischemic preconditioning (IPC) in diabetic rat hearts. METHODS: The diabetic rat model was made with alloxan. After eight weeks, 24 rats were divided into 4 groups: non-diabetic IPC rats (group A); non-diabetic non-IPC rats (group B); diabetic IPC rats (group C); diabetic non-IPC rats (group D). ECGⅡ lead, left ventricular development pressure (LVDP), and first derivative of LVDP ~(?dp/dt_~max ) were recorded. Myocardial phosphorylation of extracellular signal regulated kinases1/2 (ERK1/2) was detected by Western-blotting. RESULTS: (1) The ventricular arrythmia score was significantly lower in group A than that in group C (P
9.Expression and significance of SUFU in pancreatic carcinoma
Yunfeng WANG ; Xiaohua MAN ; Jun GAO ; Yiqi DU ; Yanfang GONG ; Zhaoshen LI ; Jing JIN ; Hongyu WU
Chinese Journal of Pancreatology 2010;10(6):418-420
Objective To investigate the expression of SUFU protein in human pancreatic carcinoma tissues and to explore the relationship between the expression of SUFU protein and the clinicopathologic parameters.Methods The expression of SUFU protein in 28 samples of pancreatic cancer tissues and 20 adjacent normal pancreatic tissues and 4 normal pancreatic tissues was detected by immunohistochemical method.And the relationship between the expression of SUFU protein and the clinicopathologic parameters were determined.Results SUFU protein was positively expressed in cytoplasm and nucleus of pancreatic carcinoma cells, while it was not expressed in the duct, acinar and islet of tumor-adjacent tissues and normal pancreatic tissues, and difference was statistically significant (P < 0.05).The high SUFU protein expression was related to the clinical stage ( P < 0.05 ), but not the age, gender, tumor location , tumor size, lymph node metastasis and liver metastasis.Conclusions SUFU protein was highly expressed in pancreatic cancer, and SUFU may play certain role in the pathogenesis of pancreatic cancer.
10.Feasibility study of induction of experimental chronic pancreatitis with L-arginine
Xiaohua MAN ; Hang ZHAO ; Kequn XU ; Yanfang GONG ; Jun GAO ; Yiqi DU ; Aifang XU ; Zhaoshen LI
Chinese Journal of Pancreatology 2009;9(2):109-111
Objective To investigate the feasibility of induction of experimental chronic pancreatitis rat model with L-arginine.Methods Animals were randomly divided into control group,arginine 12 h group,arginine 24 h group and arginine 7 d group with 10 rats in each group.L-arginine solution was intraperitoneally injected twice with an interval of 1 h.Serum amylase and glucose levels at corresponding time points were detected and histopathological scores of pancreas were evaluated.Collagen in pancreas was stained with Van Gieson method.Results Serum amylase levels were (1 634±890 ) U/L,( 3 872±2 676 ) U/L,( 3 307±2 197)U/L and (1 561±304) U/L in control group,arginine 12 h group,arglnine 24 h group and arginine 7 d group,respectively.The serum amylase level in arginine 7 d group was significantly lower than those in arginine 12 h group and arginine 24 h group (P < 0.05 ).There was no significant difference in serum glucose level among all the groups.Histopathological scores were 0.8±0.4,5.1±2.6,6.5±2.2 and 4.5±1.6,respectively.The histopathological score of arginine 7 d group was significantly lower than those in arginine 24 h group (P < 0.05 ).Obvious collagen could be found in pancreatic parenchyma in arginine 7 d group,while little collagen was found in pancreatic tissue in control,arginine 12 h and arginine 24 h groups.Conclusions Injection of L-arglnine induced fibrosis in pancreatic parenchyma and proliferation of tubular complex 7 days later,and it could be used for chronic pancreatitis model induction.