1.Effect of ischemic postconditioning on endoplasmic reticulum stress during cerebral ischemia/reperfusion in rats
Yajing YUAN ; Jincheng LI ; Qulian GUO
Chinese Journal of Anesthesiology 2014;34(9):1136-1139
Objective To evaluate the ischemic postconditioning on endoplasmic reticulum stress during cerebral ischemia/reperfusion (I/R) in rats.Methods Ninety adult male Sprague-Dawley rats,aged 350-450 g,were randomly divided into 3 groups (n =30 each) using a random number table:sham operation group (S group),I/R group,and ischemic postconditioning group (group P).Focal cerebral I/R was induced by electrocoagulation of left middle cerebral artery and 30 min occlusion of bilateral common carotid arteries followed by reperfusion.In group P,bilateral common carotid arteries were subjected to 3 cycles of 30 s reperfusion and 10 s ischemia at the beginning of reperfusion.At 24 h of reperfusion,neurological deficit was scored,and cerebral infarct size was detected by TTC staining.At 6,12 and 24 h of reperfusion,the expression of glucose-regulated protein 78 (GRP78),C/EBP homologous protein (CHOP) and caspase-12 in the ischemic area were measured (using immunohistochemistry).The neuronal apoptosis in the ischemic area was detected by TUNEL.Results Compared with S group,the neurological deficit score was significantly increased,cerebral infarct size was enlarged,the neuronal apoptosis was increased,and the expression of GRP78,CHOP and caspase-12 was up-regulated in I/R and P groups.The neurological deficit score was significantly lower,cerebral infarct size was smaller,the expression of GRP78 was higher at 12 and 24 h of reperfusion,the neuronal apoptosis was lower at 24 h of reperfusion,and the expression of CHOP and caspase-12 was lower in group P than in group I/R.Concluion Ischemic postconditioning can inhibit neuronal apoptosis mediated by endoplasmic reticulum stress during cerebral I/R,which dose not play a leading role in cerebral protection in rats.
2.The influence of aceclofenac to cartilage cell proliferation and substrate metabolism in osteoarthritis
Xia YUAN ; Kexia CAI ; Yajing QIN ; Jie YANG ; Yuezheng HU
Chinese Journal of Biochemical Pharmaceutics 2014;37(4):128-130
Objective To invesgate the influence of aceclofenac to cartilage cell proliferation and substrate metabolism in osteoarthritis.Methods Select 65 cases of patients with osteoarthritis who were treated in Department of Rheumatism and Hematology of Affiliated Hospital of Qinghai University from September 2011 to September 2013.Patients were divided into observation group(n=35)and control group(n=30).The observation group were treated with aceclofenac,while control group were treated with ibuprofen,Compare the efficacy of patients in two group.Compare human articular cartilage cells of 5-bromodeoxyuridine(BrdU)positive rate,the content of glycogen protein(PG)and collagen type Ⅱ in two groups. Results The effective rate of observation group and control group was 74.29%and 73.33% respectively.There was significant difference between the two groups. Compared with control group,the joints pain,joints tenderness,15 meters walking time,daily activity ability(P<0.05).The incidence of adverse reactions of observation group(1 1.43%)was significantly lower than that of control group(P<0.05 ).The difference of BrdU positive rate in two groups had no statistical significance. Ⅱ type collagen content of observation group was significantly higher than that of control group (P <0.05 ).PG of observation group was significantly lower than that of control group(P<0.05).Conclusion The efficacy of aceclofenac is good and the incidence of adverse reactions is low in treatment of osteoarthritis.Aceclofenac can alleviate clinical symptoms of patients by increasing collagen type Ⅱ and decreasing PG.
3.Progress in the application of rituximab in treating primary mediastinal B-cell lymphoma
Liming XU ; Yajing YUAN ; Peiguo WANG ; Gang WU
Chinese Journal of Radiation Oncology 2017;26(5):582-587
Primary mediastinal large B-cell lymphoma (PMBCL) is morphologically similar to diffuse large B-cell lymphoma (DLBCL) and nodular sclerosis Hodgkin lymphoma.For most PMBCL patients, chemotherapy plus consolidation radiotherapy showed that the latter could improve PMBCL responsiveness and progression-free survival (PFS), and its combined use with chemotherapy demonstrated higher therapeutic efficacy.Recent clinical studies suggested that rituximab and anthracycline chemotherapy regimens could increase PMBCL treatment efficacy, reduce early treatment failure, enhance PFS and overall survival, and improve prognosis.Although rituximab combined with some high-intensity chemotherapy without radiotherapy have achieved good results, many studies still support the use of post-immunochemotherapy consolidation mediastinal radiotherapy.Based on the results of a few studies with a small sample size, patients who were assessed as complete metabolic remission by PET following high-intensity immunochemotherapy may omit consolidation radiotherapy.However, these results will need to be further confirmed by large-sample multicenter clinical trials.Consolidation radiotherapy is recommended for patients with poor prognostic factors or PET score>3.
4.Research advances in radiation?induced heart disease
Liming XU ; Xi CHEN ; Yajing YUAN ; Lujun ZHAO
Chinese Journal of Radiation Oncology 2017;26(3):358-363
Radiation?induced heart disease ( RIHD) is a common type of radiation?induced damages in chest radiotherapy. There are no obvious short?term symptoms in patients with RIHD. However, RIHD causes irreversible permanent damages to the heart over time, which undermines the quality of life. Patients with severe RIHD even have a risk of death from myocardial infarction caused by coronary atherosclerosis. This paper summarizes the research advances in epidemiology, diagnosis, mechanisms of radiation?induced injury in various parts of the heart, radiotherapy techniques, and treatment. Reduction in radiation range and dose, early diagnosis, and early treatment are recommended for patients to reduce heart injury and improve the quality of life.
5.Effects of remote ischemic-postconditioning on global cerebral ischemia-reperfusion injury in rats
Bei PENG ; Qulian GUO ; Zhijing HE ; Zhi YE ; Yajing YUAN ; Na WANG ; Pingping XIA
Chinese Journal of Anesthesiology 2011;31(9):1124-1128
Objective To investigate the effects of remote ischemic postconditioning (RIPoC) on global cerebral ischemia-reperfusion (I/R) injury in rats.Methods One hundred and twenty-eight male adult SD rats weighing 200-250 g were randomly divided into 4 groups ( n =32 each):sham operation group (group S),group I/R,group I/R + RIPoC and remote I/R group (group RI/R ).Global cerebral I/R was induced by four-vessel occlusion.Group I/R + RIPoC received 3 cycles of 15 min reperfusion followed by 15 min ischemia in bilateral femoral arteries at the beginning of cerebral reperfusion.The rats were sacrificed at 24 and 48 h of cerebral reperfusion,and brains were removed for determination of neuronal apoptosis (by TUNEL method) in hippocampal CA1 region and the parietal cortex,Bcl-2 and Bax expression (by Western blot) in hippocampal CA1 region.The superoxide dismutase (SOD) and catalase (CAT) activity and malondialdehyde (MDA) content in hippocampal CA1 region and the parietal cortex were also measured at 48 h of cerebral reperfusion.Morris water maze task was used to test the learning and memory function at 4 d of cerebral reperfusion,and the rats were sacrificed at 7 d of cerebral reperfusion,and brains were removed for determination of neuronal density in hippocampal CAl region and the parietal cortex.Results Cerebral I/R significantly increased the number of apoptotic neurons and MDA content,upregulated Bcl-2 and Bax expression,decreased neuronal density,SOD and CAT activity and learning and memory function in group I/R as compared with group S.RIPoC significantly attenuated these cerebral I/R-induced changes.Conclusion RIPoC could protect brain against global cerebral I/R-induced injury,and the mechanism may be related to inhibiting lipid peroxidation,regulating the balance between Bcl-2 and Bax and inhibiting apoptosis.
6.Study on changes and influence of humoral immunity in premature infants
Chunhong WANG ; Jianhe WEI ; Zhenkui LIU ; Peixia XIAO ; Yajing ZHANG ; Erwei YUAN
Chongqing Medicine 2013;(26):3112-3114
Objective To study changes of humoral immunity of the premature infants in different pathological conditions and detect the reason of the deficiency of humoral immunity in premature infants .Methods Two hundred and forty-six prematur were enrolled and 30 healthy neonates were selected as control group .The percentages of IgG ,IgA ,IgM and comp lement C3 ,C4 were detected by full automatic biochemical analyzer .Results The results showed that IgG ,IgM ,IgA ,C3 and C4 in the premature in-fants were lower than those in the normal term infants and there was a highly significant difference with the decrease of fetal age . IgG ,IgM ,IgA ,C3 and C4 of the group of the premature infants ranging from 32 to 36 weeks had reduced in different degree ,rela-tive to the groups of BW <2 000 g ,hypertension during pregnancy ,cesarean section(P<0 .05) .Conclusion The results showed that function of humoral immunity in the premature infants was depressed and low gestational age ,low birth weight ,cesarean sec-tion and hypertension during pregnancy may be the leading cause of the deficiency of humoral immunity .
7.Myocardial response to ischemia reperfusion injury in rats after X-ray irradiation
Jun WANG ; Yajing WU ; Yuan WANG ; Sheng WANG ; Feng CAO ; Yi WANG ; Yin GUO
Chinese Journal of Radiation Oncology 2016;25(6):640-645
Objective To observe myocardial tolerance to ischemia/reperfusion (I/R) injury in rats after exposure to X-ray irradiation.Methods Twelve male rats were randomly divided into control group and radiation group.The rat model of radiation-induced heart disease was established in the radiation group by precordial irradiation with 20.0 Gy of 6 MV X-ray in a single fraction.At 14 days after model establishment,the Langendorff perfusion technique was performed in the two groups and the cardiac parameters including left ventricular developing pressure (LVDP),left ventricular end diastolic pressure (LVEDP),maximal rate of left ventricular pressure rise/fall (+/-LVdp/dtmax),and coronary flow (CF)were recorded.Myocardial infarct size after I/R was compared between the two groups by 2,3,5-triphenyltetrazolium chloride staining.Results After 30 minutes of ischemia and 60 minutes of reperfusion,the irradiation group had a significantly slower CF than the control group (5.64±0.35 vs.8.38±0.52 ml/min,P=0.002).Moreover,the irradiation group had substantially poorer recovery of cardiac function in isolated hearts compared with the control group,as shown by a significantly reduced LVDP (25.4±2.31 vs.52.76±2.76 mm Hg(1 mm Hg=0.133 kPa),P=0.000),significantly reduced+/-LVdp/dtmax(547.04±78.74 vs.1 100.05±83.35 mm Hg(1 mm Hg=0.133 kPa)/s,P=0.001;-408.81±56.74 vs-813.62±73.82mm Hg(1 mm Hg =0.133 kPa)/s,P=0.002),and a significantly increased LVEDP (85.29±4.61 vs.65.65±3.65 mm Hg (1 mm Hg =0.133 kPa),P=0.012).X-ray irradiation induced a significantly increased percentage of myocardial infarct size in rats (44.67%±0.95% vs.30.46%±0.96%,P=0.000).Conclusions X-ray irradiation can induce coronary injury,reduce myocardial tolerance to I/R injury,and increase myocardial infarct size after I/R in rats.
8.Generation and characterization of monoclonal antibody against glycoprotein VP1 encoded by caprine enterovirus CEV-JL14
Haibing LU ; Mingyue WANG ; Lisai ZHU ; Yajing LIU ; Changming GUO ; Qun ZHANG ; Yue YUAN ; Changchun TU ; Xinping WANG
Chinese Journal of Veterinary Science 2017;37(8):1468-1472
VP1 gene encoded by the newly identified caprine enterovirus CEV-JL 14 was amplified and cloned to prokaryotic expression vector pGEX-4T-1.Recombinant GST-VP1 fusion protein was expressed,purified,emulsified with Freund's complete adjuvant and used to immunize the BALB/c mice following a standard procedure.The spleen cells from immunized mice were collected and fused with myeloma cells after its antibody titer reached over 104 times detected by indirect ELISA.Hybridoma cell clones secreting monoclonal antibodies against VP1 were screened and their stability and specificity were further determined.The identified hybridoma cells were injected to mice intraperitoneally and ascites were collected at 7 DPI.Isotypes of the monoclonal antibodies against the recombinant VP1 protein were characterized to be either IgG1 or IgG2b,which showed a high specificity for detection of caprine enterovirus antigens by immunoperoxidase monolayer assay,thus laying a solid basis for future study related to viral pathogenesis,detection and diagnostics for caprine enterovirus infection.
9.PolyphyllinⅠ alleviates myocardial ischemia/reperfusion injury via nuclear factor-κB signal pathway
Liu YANG ; Yajing YUAN ; Yue WU
Chinese Critical Care Medicine 2019;31(6):746-749
Objective To investigate the protective effect of PolyphyllinⅠ(PPI) on myocardial ischemia/reperfusion (I/R) injury in rats and its mechanism. Methods The 6-month-old Sprague-Dawley (SD) rats were divided into sham operation group (Sham group), I/R model group, and low, medium, high dose PPI groups according to the random number table method, with 10 in each group. The rat myocardial I/R model was prepared by ligating the left anterior descending branch of the coronary artery by 30 minutes and reperfusion by 120 minutes. Sham group was exposure to open chest without ligation. Low, medium, high dose PPI groups were injected with PPI 75, 150, 300 mg·kg-1·d-1 in front of the film for 4 weeks; dimethyl sulfoxide (DMSO) was gastric infused in the I/R model group. After the end of reperfusion, the myocardial infarction area (IA) was determined by triphenyltetrazole (TTC) and Evans blue (EB) staining;the apoptosis of myocardial cells was detected by TdT-mediated dUTP nick end labeling stain (TUNEL); the expressions of apoptosis related protein (Bax, Bcl-2), and cytoplasmic and nucleus expressions of P65 in nuclear factor-κB (NF-κB) signal pathway were detected by Western Blot. Results Compared with the Sham group, the myocardial IA was significantly increased in the I/R model group, the apoptosis rate of myocardial cells was significantly increased, the expression of Bcl-2 was significantly decreased, and the expression of Bax was significantly increased, and the intranuclear transfer of P65 was significantly increased. Compared with the I/R model group, low, medium and high dose PPI pretreatment could significantly reduce the myocardial IA [(21.6±0.9)%, (14.3±1.6)%, (15.0±0.8)% vs. (29.6±1.4)%], the apoptosis rate of myocardial cells was significantly decreased [(38.6±1.9)%, (24.3±2.6)%, (26.3±2.8)% vs. (56.8±2.4)%], the protein expression of Bcl-2 was significantly increased, while the protein expression of Bax was significantly decreased (Bcl-2/GAPDH: 0.24±0.07, 0.36±0.02, 0.34±0.09 vs. 0.13±0.04; Bax/GAPDH: 0.39±0.10, 0.21±0.08, 0.23±0.06 vs. 0.53±0.12); and P65 nuclear transfer was significantly decreased after middle and high dose PPI pretreatment [nuclear P65/Histone 3: 0.49±0.09, 0.51±0.06 vs. 0.83±0.11; cytoplasmic P65/GAPDH: 0.31±0.03, 0.30±0.05 vs. 0.22±0.07], with statistically significant differences (all P < 0.05). However, there was no significant difference in each index between the medium and high dose PPI groups (all P > 0.05). Conclusion PPI alleviates myocardial I/R injury in rats via NF-κB signal pathway, and the PPI effect of 150 mg·kg-1·d-1 is most especially significant.
10.Establishment of an immortalized mouse mammary epithelial cell fines
Yingchao ZHANG ; Haibing LU ; Yu WANG ; Yajing LIU ; Yue YUAN ; Xinping WANG
Chinese Journal of Endocrine Surgery 2019;13(5):364-367
Objective To provide experimental materials for exploring the function of breast cancer susceptibility gene 2(BRCA2) gene by establishing of an mouse immortalized mammary gland epithelial cell line as a cell model system.Methods In this study,the primary mouse mammary epithelial cells were isolated and purified by enzyme digestion and differential centrifugation.The resulting primary mouse mammary gland epithelial cells were transducted with lentivirus expressing human telomerase (hTERT) gene,and screened by hygromycin B.The surviving epithelial cells were further characterized by morphology observations and cytokeratin marker detection.Results Immortalized mouse mammary epithelial cells were obtained by infection of MMECs with lentivirus expressing human telomerase(hTERT) gene,screened by hygromycin B.Morphology observations and detection of the cytokeratin marker showed the immortalized MMECs had a typical morphology of luminal epithelial cells with strong expression of cytokeratin 14.Conclusion An immortalized mouse luminal epithelial cell line is successfully established with an uniform morphology,which provides a cell model system for the future exploration on BRCA2-related tumorigenesis.