1.An ELISA Based on a Truncated Soluble ORF2 Protein for the Detection of PCV2 Antibodies in Domestic Pigs
Shuanghui YIN ; Shunli YANG ; Hong TIAN ; Jinyan WU ; Youjun SHANG ; Xuepeng CAI ; Xiangtao LIU
Virologica Sinica 2010;25(3):191-198
Postweaning multisystemie wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA)based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore ,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.
2.Clinical observation of vitrectomy combined with internal limiting membrane peeling and scleral shortening for myopic foveoschisis
Yawei XIAO ; Yuhua HAO ; Qingli SHANG ; Zhibo YIN ; Yaqing YANG ; Cunxi YE
Chinese Journal of Ocular Fundus Diseases 2017;33(4):373-377
Objective To observe the efficacy ofvitrectomy combined with internal limiting membrane (ILM) peeling and scleral shortening for myopic foveoschisis (MF).Methods Prospective and non-randomized concurrent control study.A total of 35 MF patients (35 eyes) were enrolled in this study.The patients were divided into 2 groups according to surgery,including group A (18 eyes) and group B (17 eyes),all received vitrectomy combined with ILM peeling,but group A also received scleral shortening.The best corrective visual acuity (BCVA) examination using the Snellen vision chart was converted to the minimum resolution logarithm (logMAR).Ocular axis length (AL) was measured by Zeiss IOL-Master or A-scan ultrasound (Quantel Medical,France).The maximal value of retinal foveoschisis (MxFT) was measured by frequency-domain optical coherence tomography (Heidelberg,Germany).Multifocal electroretinogram (mfERG) responses were obtained with the RETIscan system (Roland Consult,Gemany).There was no statistically significant difference between the two groups (P>0.05) in age (t=0.460),AL (t=1.520),diopter (t=0.020),logMAR BCVA (t=-2.280),MxFT (Z=-4.179) and b-wave ERG amplitude (Z=-0.198).The changes of BCVA,AL,MxFT and b wave amplitude were followed-up for 3-12 months.Results At the last follow-up,the height of MF was decreased in 18 eyes of group A,and MF was completely disappeared in 4 eyes.The logMAR BCVA (t=7.272,5.951),MxFT (Z=-3.724,-3.622) and b-wave ERG amplitude (Z=-3.223,-3.243) in both groups A and B were statistically improved (P=0.000,0.000,0.000,0.000,0.001,0.001) compared to pre-operational results.There was significant difference of logMAR BCVA (t=-2.280) and MxFT (Z=-4.179) between the two groups (P=0.029,0.000).But there was no significant difference in the amplitude of b-wave (Z=-0.198,P=0.843).The AL in group A was shortened after surgery,the difference was statistically significant (t=10.017,P=0.000).During the follow-up,there was no ocular hemorrhage,endophthalmitis and other complications.Conclusion PPV combined with ILM peeling and scleral shortening can shorten AL significantly for MF patients,and gain relative normal anatomical structure of the fovea,thus improve the vision.
3.Comparisons of perioperative complications and traumatic stress response between thoracoscopic radical esophagectomy and open radical esophagectomy
Yuxuan WANG ; Qiang LI ; Zhijie SHANG ; Hongfei YIN ; Gengpu YANG
Journal of Chinese Physician 2019;21(1):28-30,35
Objective To compare the incidence of perioperative complications and the degree of traumatic stress reaction between thoracoscopic radical esophagectomy and open radical esophagectomy for esophageal cancer.Methods 97 patients with esophageal cancer in our hospital were divided into group A (thoracoscopic esophagectomy,n =51) and group B (open esophagectomy,n =46) according to the operation method.The group A was given thoracoscopic radical resection of esophageal cancer,while the group B was given open radical resection of esophageal cancer.The operative indexes (intraoperative blood loss,intraoperative fluid volume,operative time,incision length),the incidence rate of perioperative complications,body traumatic stress response indexes [white blood cell count (WBC),serum C-reactive protein (CRP),free thyroxine (FT4)] and tumor markers [squamous cell carcinoma antigen (SCC),carbohydrate antigen 125 (CA125)] were compared between the two groups.Results The group A had less intraoperative blood loss,intraoperative fluid volume and shorter operative time and incision length compare with group B (P ≤ 0.05).The incidence rate of perioperative complications in group A was lower than that in group B (17.65% vs 36.96%) (P ≤ 0.05).There was no significant difference in the levels of WBC,CRP and FT4 between the two groups at the same time (P > 0.05),so as the levels of SCC and CA125 (P > 0.05).Conclusions Compared with open radical esophagectomy,the thoracoscopic esophagectomy had lower incidence of perioperative complications,but there was no significant difference in the degree of traumatic stress reaction between the two methods.
4.Relationship between the polymorphism of lipoprotein lipase gene and serum lipid levels in the Guangxi Hei Yi Zhuang population
Yong WANG ; Rui-Xing YIN ; Guang-Qin CHEN ; Wei-Xiong LIN ; De-Zhai YANG ; Shang-Ling PAN ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
0.05 for all).Conclusions The allelic frequencies of LPL gene at Pvu Ⅱ locus in Hei Yi Zhuang were different from those in Han,but the genotypie frequencies in Hei Yi Zhuang were not different from those in Han.There was no significant correlation between the polymorphism of LPL gene at Pvu Ⅱ site and the serum lipid levels in two ethnic groups.
5.Expression of dynamin-1 and phosphor-dynamin-1 in the hippocampus of children and rats with mesial temporal lobe epilepsy.
Ci-Liu ZHANG ; Fei YIN ; Zhi-Quan YANG ; Fang HE ; Chen CHEN ; Shang-Jun JIANG ; Jing PENG
Chinese Journal of Contemporary Pediatrics 2013;15(2):133-137
OBJECTIVETo observe the expression of dynamin-1 and phosphor-dynamin-1 in the hippocampus of children and rats with mesial temporal lobe epilepsy (MTLE) and to investigate the roles of dynamin-1 and phosphor-dynamin-1 in the development of MTLE.
METHODSMale Sprague-Dawley rats (aged 25 days) were randomly divided into acute control (AC), acute seizure (AS), latent control (LC), latent seizure (LS), chronic control (CC) and chronic spontaneous seizure (CS) groups. Lithium chloride-pilocarpine was used to induce a rat model of MTLE. The hippocampus samples of 5 children with a pathologically confirmed hippocampal sclerosis who received surgical operation were collected as a human model (HM) group, and the hippocampus samples of 4 dead children (without organic lesion of the hippocampus) were collected by autopsy as a human control (HC) group. The expression of dynamin-1 and phosphor-dynamin-1 in the hippocampus of children and rats with MTLE was measured by Western blot and immunohistochemistry.
RESULTSThe Western blot showed that the expression of phosphor-dynamin-1 was significantly lower in the AS and CS groups than in the corresponding control groups (AC and CC groups) (P<0.05). The expression of phosphor-dynamin-1 was significantly lower in the HM group than in the HC group (P<0.05). There were no significant differences in the expression of dynamin-1 among the AS, LS and CS groups and between the HM and HC groups (P>0.05). The immunohistochemical results showed that phosphor-dynamin-1 was highly expressed in the cytoplasm of hippocampal neurons of AC, CC and HC groups, but its expression was significantly reduced in the AS, CS and HM groups (P<0.05).
CONCLUSIONSThe expression of phosphor-dynamin-1, not dynamin-1, is downregulated in the hippocampus of children and rats with MTLE during seizures, which suggests that the phosphorylation/dephosphorylation of dynamin-1 may be involved in the development of MTLE.
Animals ; Blotting, Western ; Child ; Dynamin I ; analysis ; metabolism ; Epilepsy, Temporal Lobe ; metabolism ; Female ; Hippocampus ; chemistry ; metabolism ; Humans ; Immunohistochemistry ; Male ; Phosphorylation ; Rats ; Rats, Sprague-Dawley
6.Effect of inhibiting endoxin by antidigoxin antiserum on myocardial ischemia/reperfusion injury in rats.
De-Guo WANG ; He-Gui WANG ; Yong-Sheng KE ; Shang-Yin YANG
Chinese Journal of Applied Physiology 2007;23(1):9-12
AIMTo study the effect of antidigoxin antiserum on oxygen stress induced by myocardial ischemia/reperfusion (MI/R) injury in rats.
METHODSSprauge Dawley rats were submitted to ligate left anterior descending coronary artery 30 min followed by 45 min reperfusion. Experiment animals were randomly divided into seven groups including sham group, MI/R group, normal salina group, verapamil group and three antidigoxin antiserum groups from low to high dose. The left ventricular myocardial tissue sample of ischemia were processed and measured the level of endoxin and malondialdehyde (MDA), the activities of Na+, K(+) -ATPase and superoxin dismutase (SOD). The myocardia morphology was observed.
RESULTSThe levels of endoxin and MDA increased and the activities of Na+, K(+) -ATPase and MDA were inhibited significantly in MI/R and saline groups. Including verapamil group in comparison to MI/R and saline groups, MDA level decreased and SOD activities partly reserved, meanwhile, only in three antidigoxin antiserum groups, the myocardial endoxin level was remarkably decreased, Na+, K(+) -ATPase activities were drastically increased. The myocardial histological morphology was significantly improved.
CONCLUSIONAntidigoxin antiserum, an endoxin mutual clone antibody, had the effect of attenuating the damage of oxygen free radicals induced by MI/R via to antagonizing the inhibition effect of endoxin on myocardial membrane Na+, K(+) -ATPase activities.
Animals ; Cardenolides ; antagonists & inhibitors ; Digoxin ; pharmacology ; Immune Sera ; pharmacology ; Malondialdehyde ; analysis ; Myocardial Reperfusion ; Myocardial Reperfusion Injury ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Saponins ; antagonists & inhibitors ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism
7.Retros Flt-1 decelerates the growth of a murine experimental osteosarcoma.
Xiao-tang XIN ; De-zhen YIN ; Hai LAN ; Cong CHEN ; Bo LIU ; Shang-you YANG
Chinese Journal of Surgery 2011;49(8):746-751
OBJECTIVETo examine the influence of vascular endothelial growth factors (VEGF) in controlling the growth of an experimental osteosarcoma in mice by performing retrovirus-mediated sFlt-1 gene modification.
METHODSFrom March to October 2010 human osteosarcoma G-292 cells were in vitro infected with retroviral vectors encoding soluble Flt-1 or LacZ gene before transplanted into proximal tibiae of immune deficient SCID mice to establish experimental orthotopic osteosarcoma. Daily observation and biweekly microCT were performed to monitor tumor development and progression till sacrifice at 8 weeks after tumor cell inoculation for histological and molecular analyses.
RESULTSSuccessful transgene expression was confirmed in the culture media of sFlt-1 transduced G-292 cells using ELISA, and with positive X-gal staining of the LacZ transduced cells. Noteworthy tumors were grown in all mice on the tibiae receiving G-292 cell inoculation, with clear detection on microCT images starting 2 weeks after inoculation. Over the time period, tumors derived from sFlt-1 transduced G-292 cells were distinctively smaller in size compared to the ones from wide-type G-292 and G-292-LacZ cells. Histology showed typical osteosarcoma characteristics including severe cellular pleomorphism, bone erosions, and neo-vascularization. Real-time polymerase chain reaction indicated significantly higher sFlt-1 expression in sFlt-1 transduced groups than the wild-type G-292 or LacZ treated groups. Strong expression of oncogenes c-myc and c-fos were also obvious, along with the expression of VEGF in the primary tumor tissue.
CONCLUSIONRetrovirus-mediated sFLT-1 gene modification decelerates the osteosarcoma tumor growth in this murine model.
Animals ; Bone Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Genetic Vectors ; Humans ; Lac Operon ; Mice ; Mice, SCID ; Neovascularization, Pathologic ; metabolism ; pathology ; Osteosarcoma ; genetics ; metabolism ; pathology ; Retroviridae ; genetics ; Transgenes ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism
8.Relationship between screw numbers and severity of tibial bone defect in primary total knee arthroplasty.
Chong ZHENG ; Yong-gang ZHOU ; Hai-yang MA ; Zhuo ZHANG ; Hua-hao FU ; Wen-ming WU ; Shang PIAO ; Yin-qiao DU ; Sen WANG
China Journal of Orthopaedics and Traumatology 2016;29(5):415-420
OBJECTIVETo summarize experience of using screws and cement to rebuild tibial bone defect in primary total knee arthroplasty (TKA) and to discuss the relationship between the number of required screws and the severity of tibial bone defects.
METHODSFrom July 2009 to May 2015, 34 patients (40 knees) with varus knees underwent TKA, and the screw and cement technique was used to rebuild medial tibia plateau during operation. There were 8 males (8 knees) and 26 females (32 knees), and the average age was (65.00 +/- 7.25) years old (ranged,55 to 82 years old). One to 6 screws were used in each case. Extension stems were used in 2 cases (4 and 5 screws was used respectively). The area percentages of the bone defects measured as defect area/tibia plateau area, depth of each defect, the number of screws needed in each case, were all used to determine the relationship between the number of screws and the area percentage in certain depth of bone defect by statistic methods, as well as the relationship between screw number and defect depth.
RESULTSAll the patients were followed up and the average duration was 24 months (ranged, 1 to 72 months). The average preoperative HSS score was 43.33 +/- 6.11 (ranged, 32 to 51 scores). Whereas the average postoperative HSS score was 92.15 +/- 4.64 (ranged,83 to 96 scores). The preoperative individual scores including pain, function, activity, nuscle strength, flexion deformity and stability were all improved compared with preoperation,and the differences were statistically significant. All the patients received normal alignment postoperatively, femoraltibial angle was improved from (167.00 +/- 6.39) degrees preoperatively to (175.00 +/- 2.69) degrees postoperatively, the tibial angle was improved from (78.09 +/- 4.51) degrees preoperatively to (88.75 +/- 1.24) degrees postoperatively. Both area percentage and depth of bone defect in a fitting Ologistic model had a significant statistical relationship with the screw number, and a rectangular coordinate system could be formed according to the relationship.
CONCLUSIONScrews and cement technique is a simple, safe and convenient method to rebuild tibial bone defects in primary TKA and its short-term and midterm effect are both reliable. During opera- tion, according to the rectangular coordinate system, the screw number needed in the operation can be inferred form th area and depth of tibia defect, which could have a guiding function in surgery.
Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; instrumentation ; methods ; Bone Screws ; utilization ; Female ; Fracture Fixation, Internal ; Humans ; Knee Injuries ; surgery ; Knee Joint ; surgery ; Male ; Middle Aged ; Tibia ; surgery
9.New classification of Crowe type IV developmental dysplasia of the hip.
Hai-yang MA ; Yong-gang ZHOU ; Chong ZHENG ; Wen-zhe CAO ; Wang SEN ; Wen-ming WU ; Shang PIAO ; Yin-qiao DU
China Journal of Orthopaedics and Traumatology 2016;29(2):119-124
OBJECTIVETo compare differences between Crowe IV developmental dysplasia of the hip (DDH) with secondary acetabulum and Crowe IV DDH without secondary acetabulum,and determine whether it is necessary to divide Crowe IV DDH into two subtypes.
METHODSFrom June 2007 to May 2015,145 hips of 112 Crowe N patients who underwent total hip arthroplasty (THA) using S-ROM stem were divided into two groups: secondary acetabulum formaton group (group A) and no secondary acetabulum formaton group (group B). In group A,there were 12 females, 96 males,with an average age of (39.38 ± 11.19) years old. In group B, there were 2 females, 35 males, with an average age of (38.19 ± 10.92) years old. All the patients were evaluated by using Harris Hip Score. Radiographic evaluations were made preoperatively and during follow up. The differences between two groups were compared on dislocation height, canal flare index (CFI), subtrochanteric shortening osteotomy (SSTO) usage, pre- and post-operation Harris scores, complications.
RESULTSThe dislocation height for group A was (4.74 ± 1.57) cm, while the dislocation height for group B was (3.12 ± 1.15) cm. Significantly difference was detected between two groups. The CFI for group A was 2.69 ± 0.68, while the CFI for group B was 3.42 ± 0.79, and the significantly difference was detected between two groups. Harris scores were totally improved from 58.18 ± 15.67 preoperatively to 91.20 ± 3.79 post-operatively and the difference was significant. Pre-operative Harris scores was 58.1 ± 15.3 in group A, 58.3 ± 16.9 in group B. Post-operative Harris scores was 91.0 ± 4.1 in group A, 91.0 ± 5.1 in group B. No significant difference was found on Harris scores between A and B preoperatively and post-operatively. Complications of 4 cases peri-prosthesis fracture, 4 cases dislocation and 4 cases nerve injury occur in group A; While only one case dislocation and one case nerve injury occur in group B. No statistical significance was detected.
CONCLUSIONCrowe IV DDH with secondary acetabulum is significantly different from Crowe IV DDH without secondary acetabulum on dislocation height and femoral morphology, which causes the different selections of surgical techniques (SSTO usage or not). These important differences in fundamental parameters indicate the necessity to further divide Crowe IV DDH into IVA and IVB two subtypes.
Adolescent ; Adult ; Aged ; Female ; Hip Dislocation, Congenital ; classification ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; therapy
10.Correlation between SLAMF6 expression and perforin and granzyme B on circulating CD8 +T cells and the clinical significance in patients with severe aplastic anemia
Baojun SHANG ; Shiwei YANG ; Rongjun MA ; Xiaoli YUAN ; Li JIANG ; Pingchong LEI ; Zhongwen LIU ; Yulong LI ; Xiaoyan DONG ; Lin ZHANG ; Yin ZHANG ; Zunmin ZHU
Chinese Journal of Laboratory Medicine 2021;44(3):233-238
Objective:To explore the correlation between the expression of signaling lymphocyte activation molecule family 6 (SLAMF6) on peripheral blood CD8 +T cells and perforin and granzyme B and the clinical significance in patients with newly diagnosed severe aplastic anemia(SAA). Methods:The indicators of blood routine and bone marrow and peripheral blood samples of 32 newly diagnosed SAA patients admitted to Henan Provincial People′s Hospital from January 2016 to June 2019 were collected for retrospective analysis. Flow cytometry was used to detect the expression of SLAMF6, perforin and granzyme B on samples CD8 +T cell before therapy and 6 months after therapy (11 cases received transplantation, 21 cases received immunosuppressive therapy [IST]). Spearman correlation analysis was performed to determine the association between clinical indicators and laboratory test results. The expression of SLAMF6, perforin and granzyme B was also detected in 10 healthy people (normal group) and 13 myelodysplastic syndromes/paroxysmal nocturnal hemoglobinuria (MDS/PNH) patients (MDS/PNH group). Results:(1) At diagnosis: the expression of SLAMF6 was significantly lower in the SAA group than that in the normal group and the MDS/PNH group ([56.40±6.37]% vs [84.34±5.81]% and [82.24±4.98]% (both P<0.001]). The expression of perforin was significantly higher in the SAA group (32.73±8.46) than that in the normal control group (23.75%±5.10%), and the MDS/PNH group (26.12%±5.53%) (both P<0.05). The expression of granzyme B was also significantly higher in the SAA group (36.23%±7.94%) than that in the normal control group (21.67%±5.05%) and the MDS/PNH group (21.79%±5.10%) (both P<0.001). The expression of SLAMF6 was positively correlated with the hemoglobin ( r=0.804), and reticulocyte absolute values ( r=0.656) in peripheral blood, percentage of granulocytes ( r=0.643) and erythrocytes ( r=0.622) in bone marrow of SAA patients (all P<0.05). Expression of SLAMF6 was negatively correlated with perforin ( r=-0.792) and granzyme B ( r=-0.908) on CD8 +T cells in patients with SAA (both P<0.001). (2) After treatment: the expression of SLAMF6 in peripheral blood CD8 +T cells of 30 surviving patients was higher than pre-treatment ([79.19±12.69]% vs [56.40±6.37]%, P<0.001). The expressions of perforin and granzyme B were lower than pre-treatment level (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 11 transplanted patients was higher than before transplantation ([86.54±3.75]% vs [56.40±7.35]%, P<0.001). The expressions of perforin and granzyme B were lower than before transplantation (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 12 IST-respond patients was higher than that before treatment, while the perforin and granzyme B levels were lower than pre-treatment (all P<0.05). The post-treatment expressions of SLAMF6, perforin and granzyme B were similar as before treatment levels in 7 IST-unrespond patients (all P>0.05). Conclusion:SLAMF6 is significantly down-regulated on CD8 +T cells in newly diagnosed SAA, negatively correlated with the effective factors of CD8 +T cells, which might participate in the immune regulatory of CD8 +T cells as a negative regulatory factor in patients with SAA. The SLAMF6 is significantly up-regulated after hematopoietic recovery, while there is no significant change in treatment-unrespond patients, which could thus serve as an useful diagnostic and therapeutic index of patients with SAA.