Impact of vaccination against COVID-19 on the outcomes of in vitro fertilization-embryo transfer: a retrospective cohort study / 中华医学杂志(英文版)

BACKGROUND@#Vaccination against coronavirus disease 2019 (COVID-19) has become the primary approach in the fight against the spread of COVID-19. Studies have shown that vaccination against COVID-19 has adverse effects, particularly on human reproductive health, despite the fact that vaccination rates are still on the rise. However, few studies have reported whether vaccination affects the outcome of in vitro fertilization-embryo transfer (IVF-ET) or not. In this study, we compared the outcome of IVF-ET and the development of follicles and embryos between vaccinated and unvaccinated groups.@*METHODS@#A single-center retrospective cohort study of 10,541 in vitro fertilization (IVF) cycles was conducted from June 2020 to August 2021. 835 IVF cycles with a history of vaccination against COVID-19 and 1670 IVF cycles that served as negative controls were selected and analyzed utilizing the Matchlt package of R software ( http://www.R-project.org/ ) and the nearest neighbor matching algorithm for propensity-matched analysis at a 1:2 ratio.@*RESULTS@#The number of oocytes collected in the vaccinated group and the unvaccinated group were 8.00 (0, 40.00) and 9.00 (0, 77.00) ( P  = 0.073) and the good-quality embryo rates of the two groups were 0.56±0.32 and 0.56±0.31 averagely ( P  = 0.964). Clinical pregnancy rates for the vaccinated group and unvaccinated group were 42.4% (155/366) and 40.2% (328/816) ( P  = 0.486) and biochemical pregnancy rates were 7.1% (26/366) and 8.7% (71/816) ( P  = 0.355). Two other factors were analyzed in this study; vaccination among different genders and different types (inactivated vaccine or recombinant adenovirus vaccine) showed no statistically significant effect on the above outcomes.@*CONCLUSIONS@#In our findings, vaccination against COVID-19 showed no statistically significant effect on the outcomes of IVF-ET and the development of follicles and embryos, nor did the gender of the vaccinated person or the formulation of vaccines show significant effects.

Clinical analysis of de Winter syndrome in six patients / 中国基层医药

Objective:To investigate the pathological characteristics, treatment timing, and prognosis of de Winter syndrome.Methods:Six patients with de Winter syndrome who received treatment in the Department of Cardiovascular Medicine, The First People's Hospital of Tianmen from July 2017 to September 2020 were included in this study. The clinical risk factors, characteristics of coronary artery lesions, electrocardiogram evolution, echocardiography, high-sensitivity troponin, and brain natriuretic peptide were evaluated. All patients were followed up for 12 months after discharge.Results:Among the six patients included, four patients underwent coronary angiography and percutaneous coronary intervention. Coronary angiography results showed that anterior descending artery lesions occurred in all patients, consisting of occlusion of the anterior descending artery in three patients and severe stenosis of the anterior descending artery in one patient. After surgery, TIMI3 blood flow recovered in all patients. Electrocardiogram showed anterior wall ST segment elevation in five patients, and anterior wall and inferior wall ST segment elevation in one patient. One patient refused to undergo coronary angiography and was discharged after conservative management with drugs. de Winter syndrome was not identified in time in one patient. The patient died after being admitted to the hospital through routine procedures. Five recovered patients were followed up for 12 months, consisting of one patient who was re-admitted because of heart failure, and four patients in whom no adverse events occurred.Conclusion:Identification of electrocardiogram manifestations of de Winter syndrome and implementation of coronary angiography and percutaneous coronary intervention as early as possible can substantially reduce mortality rate and improve long-term prognosis.

Clinical study on the expression of Piezo1 in small intestinal mucosa of 57 patients with Crohn′s disease / 中华消化杂志

Objective:To investigate the expression of Piezo1 in small intestinal mucosal epithelial cells of patients with Crohn′s disease (CD) and its clinical correlation with CD.Methods:From January 1st 2010 to November 30th 2020, the clinical data including age, gender, disease location and biological behavior, etc of 57 patients with CD (CD group) who underwent surgery at The First Affiliated Hospital of Anhui Medical University were retrospectively. And at same time the normal samll intestinal epithelial tissues of 10 healthy individuals who underwent colonoscopy were collected as the healthy control group. The expression of Piezo1 in small intestinal epithelial cells of CD patients with different disease sites, biological behavior and disease activity were detected by immunofluorescence staining and hematoxylin-eosin staining. The histological score system and intestinal fibrosis score were used to analyze the inflammation and fibrosis of the intestinal tissues of patients with CD. Semi-quantitative analysis of Piezo1 in small intestinal epithelial cells was analyzed by ImageJ software. And the correlation between Piezo1 expression and clinical characteristics and pathological features of small intestine was also analyzed. Independent sample t test and analysis of variance were used for statistical analysis. Results:In CD group, there were 37 males (64.9%) and 20 females (35.1%). The age was (39.1±14.2) years old, ranged from 18 to 71 years old, and the average duration of the disease was (26.5±24.1) months. There were 29 cases (50.9%)of ileal type, 26 cases (45.6%) of ileocolonin type and 2 cases (3.5%) of colonic type. There were 12 cases (21.1%) of non-penetrating non-stenotic type, 31 cases (54.4%) of stenotic type and 14 cases (24.6%) of penetrating type. There were 47 cases (82.5%) with moderate activity and 10 cases (17.5%) with severe activity. There were 17 cases (29.8%) of moderate intestinal inflammation, 40 cases (70.2%) of severe intestinal inflammation. The score of intestinal fibrosis in six cases (10.5%) was 1, 28 cases (49.1%) was 2, 18 cases (31.6%) was 3, five cases was 4. The relative expression level of Piezo1 in intestinal mucosal epithelial cells of CD group was higher than that of healthy control group (12.9±4.6 vs. 8.5±1.1), the relative expression of Piezo1 in intestinal mucosal epithelia cells of stenotic type and penetrating type CD patients were both higher than that of non-penetrating and non-stenotic CD patients (12.6±3.8 and 9.8±2.4 vs. 6.0±1.3), and the differences were all statistically significant ( t=3.00, -3.66 and -3.32, all P<0.01). The relative expression of Piezo1 in small intestinal epithelial cells of CD patients with severe intestinal inflammation was higher than that of CD patients with moderate intestinal inflammation (13.1±4.0 vs. 9.7±3.1), and the difference was statistically significant ( t=-2.65, P<0.05). The relative expression levels of Piezo1 in small intestinal epithelial cells of patients with intestinal fibrosis score of 4, 3, 2 and 1 were 17.6±5.2, 12.6±1.7, 9.1±2.1 and 5.8±1.1, respectively; the relative expression levels of Piezo1 in intestinal epithelial cells of patients scored 4 were higher than that of patients scored 3, 2 and 1, and that of patients scored 3 was higher than patients scored 2 and 1, and that of patients scored 2 was higher than that of patients scored 1, and the differences were all statistically significant ( t=-2.98, -5.10, -3.84, 4.60, 6.55 and 2.56, all P<0.05). The relative expression of Piezo1 in intestinal mucosal epithelial cells was related to the severity of intestinal inflammation and fibrosis. The more severe the intestinal inflammation and fibrosis, the higher the relative expression of Piezo1 in intestinal mucosal epithelial cells. Conclusions:The relative expression of Piezo1 in small intestinal epithelial cells is related to the biological behavior and the severity of intestinal inflammation and fibrosis of CD. It is speculated that the expression of Piezo1 in small intestinal epithelial cells may be clinically related to the process of intestinal wall fibrosis in CD to some extent, however whether it plays an important role in the process of intestinal wall fibrosis in CD and its specific mechanism need to be further studied.

Effect of fucoidan on autophagy, migration and invasion of U266 cells / 上海交通大学学报（医学版）

Objective · To investigate effect of fucoidan on autophagy, migration and invasion in human multiple myeloma U266 cells. Methods · The U266 cells treated with fucoidan were cultured in vitro. The formation of autophagosomes was observed by transmission electron microscopy (TEM). Transwell assay was used to evaluate the effect of fucoidan on migratory and invasive abilities of U266 cells. The protein levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, P62, MMP9, CXCR4, p-AKT/T-AKT, p-mTOR/T-mTOR were detected by Western blotting. MMP9 concentration in the culture medium was examined by ELISA. Results · ① Autophagosomes increased in fucoidan-treated cells compared with control group under TEM. ② Migratory and invasive abilities were inhibited by fucoidan in a dose-dependent manner, which were suppressed by chloroquine. ③ Western blotting demonstrated that expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and MMP9 increased in fucoidan-treated cells, while P62, CXCR4, p-AKT/T-AKT and p-mTOR/T-mTOR decreased compared with control group. ④ The result of ELISA showed that MMP9 concentration in the culture medium of fucoidan-treated cells significantly decreased. Conclusion · Fucoidan induces autophagy and inhibits migration and invasion in U266 cells.

Clinical expression of P53,Livin and PARP in the epithelial ovarian cancer and its impact in the chemotherapy resistance and clinical prognosis / 中国生化药物杂志

Objective To explore the clinical expression of P53, Livin and PARP in the epithelial ovarian cancer and its correlation with the chemotherapy resistance and clinical prognosis.Methods 74 specimen of epithelial ovarian cancer confirmed from January 2009 to June 2011 in our gynecology department were selected.During the follow-up visit, the subjects were divided into chemotherapy sensitivity group and chemotherapy resistance group according to the recurrence cases, the clinical expression and survival rate for two groups were compared, the influence factors of survival time were analyzed.Results The positive rate of P53, Livin and PARP for chemotherapy sensitivity group was 47.1%, 56.9%and 52.9%;the positive rate for chemotherapy resistance group was 73.9%, 95.7% and 95.7%,the diyforences were significant(P<0.05).After 1, 3 and 5 years of treatment, the survival rate for chemotherapy sensitivity group was 100.0%, 82.4% and 66.7%,The survival rate for chemotherapy resistance group was 87.0%, 26.1% and 8.7%,the diyforences were significant(P<0.05).Based on the Cox regression model, the influence factors of the patient's age, pathological differentiation degree, clinical staging and chemotherapy sensitivity were introduced.It was known that the patient's survival time was greatly influenced by clinical staging and chemotherapy sensitivity (P<0.05).Conclusion For patients with epithelial ovarian cancer, the expression of P53, Livin and PARP is correlated with chemotherapy resistance.Therefore, the clinical effect is predictable, for patients with higher expression, the personalized therapy can improve the patient's prognosis.

Effects of miR-335-5p on the osteoblast function in high glucose condition / 中华内分泌代谢杂志

Objective To investigate the effects of miR-335-5p on the proliferation and apoptosis of osteoblasts which were exposed to high glucose condition, and explore its possible molecular mechanisms. Methods MC3T3-E1 osteoblasts were divided into four groups:control group(5. 5 mmol/L glucose), high glucose group(HG group, 22. 0 mmol/L glucose), agomir-335-5p group(transfected with agomir-335-5p and exposed to 22. 0 mmol/L glucose) , and agomir negative control group( agomir NC group, transfected with agomir negative control and exposed to 22. 0 mmol/L glucose), cultured for 7 days. Cell proliferaton, cell apoptosis, expressions of miR-335-5p and dickkopfhomolog1(DKK1)mRNA,proteinlevelsofDKK1andcysteinylaspartate-specificproteinase-3(caspase-3) were detected using MTT, flow cytometry, quantitative realtime PCR and western blot, respectively. Results Compared with control group, the expression of miR-335-5p mRNA and cell proliferation in HG group were significantly decreased(P0. 05). The miR-335-5p mRNA expression and cell proliferation in agomir-335-5p group were higher than those in HG group and agomir NC group(P<0. 05). However, Cell apoptosis and the protein levels of DKK1 and caspase-3 in agomir-335-5p group were lower than those in HG group(P<0. 05). Conclusion High glucose inhibits the proliferation and induce the apoptosis of MC3T3-E1 osteoblast through decreasing the expression of miR-335-5p and subsequently increasing the DKK1 expression.

Clinical therapeutic effect of recombinant human erythropoietin on patients with heart failure compli-cated chronic kidney disease / 心血管康复医学杂志

Objective:To observe clinical therapeutic effect of recombinant human erythropoietin (rhEPO)on pa-tients with heart failure complicated chronic kidney disease.Methods:A total of 84 patients with heart failure com-plicated renal insufficiency were selected.They were divided into routine treatment group (n=42)and rhEPO group (n=42,received rhEPO based on routine treatment)according to random number table method.Echocardiographic examination results and renal function were compared between two groups after 12-week treatment.Results:Com-pared with routine treatment group,there were significant rise in left ventricular ejection fraction [LVEF,(37.2± 10.3)% vs.(45.4 ± 11.4)%]and left ventricular fractional shortening [LVFS,(19.6 ± 4.3)% vs.(24.5 ± 3.8)%],and significant reductions in left ventricular end-diastolic diameter [LVEDd,(6.12±0.67)mm vs.(5.01 ±0.54)mm],24h urinary protein [(0.76±0.1)g vs.(0.24±0.09)g],24h urine microalbumin [(319.6±39.6) mg vs.(107.3±26.7)mg],blood urea nitrogen [(10.3±1.9)mmol/L vs.(6.2±1.5)mmol/L]and serum creati-nine [(97.2±16.8)μmol/L vs.(79.3±15.7)μmol/L]in rhEPO group,P<0.01 all.Conclusion:Recombinant human erythropoietin could significantly improve heart,renal function in patients with heart failure complicated re-nal insufficiency.

The infection status and genotype distribution of high risk human papillomavirus in Chongqing women / 重庆医学

Objective To evaluate the infection status and genotype distribution of high risk human papillomavirus (HPV) in Chongqing women with different healthy status of cervix .Methods A retrospective analysis including 3 118 cases from June 2013 to March 2014 were performed ,945 of which were carried out multiplex real time PCR ,and data of cervix healthy status and HPV positive cases were subjected to chi‐square test .Results The general positive percentage of high risk HPV was 27 .49% ,16 .35%in healthy check‐up group ,and 24 .42% in cervicitis group and 39 .87% in cervical neoplasia lesion ,there was a significant difference of HPV positive ratio between these three groups(P< 0 .05) ;and HPV positive ratio was higher in older women without significant difference .In Chongqing region ,the three most common HPV types were HPV52 (21 .97% ) ,HPV16 (20 .00% ) and HPV58 (14 .75% ) .However ,the three most common HPV types in various groups are different :HPV52 (26 .20% ) ,HPV58 (14 .3% ) and HPV51 (10 .70% ) in health check‐up group ; HPV52 (21 .10% ) ,HPV58 (20 .00% ) and HPV16 (18 .90% ) in cervicitis group while HPV16 (29 .00% ) ,HPV52 (19 .80% ) and HPV58 (11 .50% ) in cervical neoplasia lesion group .The multiple infec‐tion rate of HPV was 5 .86% ,as the cervix status gets worse ,HPV concurrent infection increases ,and concurrent HPV types vary :mostly with HPV 52 in healthy check‐up group while with HPV 16 in cervical neoplasia lesion group .Conclusion The prevalence and genotype distribution of HPV in Chongqing women with different healthy status of cervix are significant difference.

Effects of renal carcinoma cell line ACHN-derived exosomes on ACHN cell proliferation and apoptosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of exosomes derived from renal cancer cell line ACHN on the proliferation and apoptosis of ACHN cells and explore the mechanism.</p><p><b>METHODS</b>Exosomes derived from ACHN cells were separated and purified by ultrafiltration and sucrose gradient centrifugation. The effects of the exosomes on the proliferation and apoptosis of ACHN cells were analyzed with CCK-8 assay and flow cytometry, respectively. The changes of mRNA and protein expressions of cyclin D1, caspase-3 were examined using RT-PCR and Western blotting, and the changes in the protein expression of p-Akt and p-ERK1/2 were detected with Western blotting.</p><p><b>RESULTS</b>Exosomes were successfully purified by ultrafiltration and sucrose gradient centrifugation. Compared with the control cells, ACHN cells treated with the exosomes showed enhanced proliferative activity with suppressed cell apoptosis. Exosomes treatment upregulated cyclinD1 mRNA and protein expression, down-regulated caspase-3 protein expression without affecting caspase-3 mRNA expression, and upregulated the expression of p-Akt and p-ERK1/2.</p><p><b>CONCLUSION</b>Exosomes can promote the growth and proliferation and inhibit the apoptosis of renal cancer cell line ACHN. Removal of the exosomes from the microenvironment of renal cancer or inhibition of its function can be new strategies for treatment of renal cancer.</p>

Effect of tetrandrine on nitroglycerin induced activation of satellite cells in trigeminal ganglia / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the effect of tetrandrine (Tet) on nitroglycerin(GTN)-induced activation of the satellite cells released inflammatory cytokines and to explore its mechanism.</p><p><b>METHOD</b>Neonatal rat satellite cells of trigeminal ganglia were cultured and separated into three groups. Group CON: the cells were normal cultured; Group TGN: the cells were cultured with 0.55 mmol x L(-1) GTN; Group Tet: the cells were treated with 0.55 mmol x L(-1) GTN and 1 x 10(-7) mol x L(-1) Tet respectively. Cell viability after GTN and Tet was detected by AlamarBlue assay. The concentration change of intracellular Ca2+ ([Ca2+]i) in single satellite cell loaded with Fluo-3/AM was determined by laser scanning confocal microscopy. NF-kappaB and IL-1beta mRNA levels were determined by FQ-PCR. Through double-immunofluorescent staining identifies satellite cells and determines the expression of NF-kappaB protein.</p><p><b>RESULT</b>Satellite cells activities decreased with GTN stimulating, but according to the viability and modality of the cells, 1 x 10(-7) mol x L(-1) Tet was the suitable prophylaxis. Tet can inhibit the elevation of cytosolic free calcium of rat satellite cell and decrease the mRNA and protein levels of NF-kappaB and the mRNA levels of IL-1beta.</p><p><b>CONCLUSION</b>Via preventing Ca2+ influxion, Tet inhibited NF-kappaB activation of satellite cell which decreased IL-1beta expression.</p>

Preliminary establish of the puberty pathological aggression animal model / 中华行为医学与脑科学杂志

Objective To establish the preferable puberty pathological aggression animal model.Methods The experimental models were established towards puberty rats by frustration test of non-reward and instigation test.Rat models were tested for specificity using open-field test,saccharine test,elevated plus-maze(EPM)and olfactory sensibility.Results ①Compared with normal aggression(52.5±5.36)and control group(8.83±1.34)in total aggressive times,the pathological aggression group(101.17±2.85)increased significantly(P0.05).However,the normal aggression group displayed obvious depressive mood.Conclusion Each behavioral index matches the criteria of pathological aggression model.Meanwhile,it also excludes other factors of disturbing the specificity of the model.It suggests this model may be preferable puberty pathological aggression animal model.

Protective Effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside on PC12 Cells Injury Induced by H2O2 / 中国康复理论与实践

@#ObjectiveTo study the protective effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(TSG) on the PC12 cells injury induced by H2O2.MethodsAll cells were divided into 5 groups: the saline control group(control group), Oxidative damage PC12 cells model group(H2O2 group) which was induced by H2O2, and TSG treated group, which oxidative damage PC12 cells model which was induced by H2O2 after given TSG 2 h at TSG 120 μg/L(TSG 120 μg/L+H2O2 group), 60 μg/L(TSG 60 μg/L+H2O2 group) and 30 μg/L(TSG 30 μg/L+H2O2 group) once. The survival rate of the cells was determined by MTT method, the content of lactate dehydrogenase (LDH) was determined by ultraviolet spectrophometry and the content of malonyldialdehyde (MDA) and superoxide dismutase (SOD) activity was measured respectively by thiobarbituric aicd and xanthine oxidese method. Immunohistochemitry method were used to detect the expression of bcl-2.ResultsTSG reduced obviously cells injury induced by H2O2. 30～120 μg/L TSG improved the cells survival rate, reduced LDH releasing and MDA content, increased SOD activity, and decreased the expression of bcl-2 in the PC12 cells injury induced by H2O2(P<0-05, P<0-01). ConclusionTSG has significantly potective effect on the PC12 cells injury induced by H2O2.

Effect of Functional Electrical Stimulation on Repairing Spinal Cord Injury / 中国医学物理学杂志

Objective:To review effects of functional electrical stimulation(FES) on paralyzed or paretic muscles,and introduce the progress of functional electrical stimulation repairing spinal cord injury.Methods:A computer-based search was conducted in CNK/and MEDILINE for articles related to functional electrical stimulation repairing spinal cord injury from November 1990 to November 2009 with the Keywords of spinal cord injury and electrical stimulation.Data were check firstly.Articles related to functional electrical stimulation repairing spinal cord injury were selected and looked for full-texts.The articles were analyzed and summarized.Results:Functional electrical stimulation (FES) can activate paralyzed or paretic muscles to generate functional or therapeutic movements with constant frequency.FES can improve the impaired motor function of muscles stimulated.Functional electrical stimulation technique could repair part of motor function following spinal cord injury.Conclusions:Functional electrical stimulation is a new and promising technique in modem rehabilitation engineering.Functional electrical stimulation (FES) can activate paralyzed or paretic muscles to generate functional or therapeutic movements.FES can improve the impaired motor function of muscles stimulated.

Effect of endoplasmic reticulum stress in renal damage caused by hyperlipidemia / 西安交通大学学报(医学版)

Objective To investigate the effect of endoplasmic reticulum stress in renal damage caused by hyperlipidemia. Methods Forty male Wistar rats were randomly divided into two groups: normal control group (NC, n=20) and high-fat group (HF, n=20). Rats in NC group were fed with normal diet while those in HF group were fed with high-fat diet. Five rats in each group were randomly chosen in week 4, 8, 12 and 18. Serum lipid, urine protein in 24 hours and the pathological changes of renal tissues were observed; the apoptosis of renal cells was detected by TUNEL staining; the expression of GRP78 protein in the kidney was examined by immunohistochemistry. The expression of GRP78 mRNA and CHOP mRNA was examined by RT-PCR. Results Compared with those in NC group, serum lipid as well as the expression of GRP78 mRNA and protein in the kidney were increased in week 4, 8, 12 and 18 in HF group(P<0.05). In contrast, urine protein in 24 hours, the apoptosis index of renal cells and the expression of CHOP mRNA were increased in week 8, 12 and 18 (P<0.05). Conclusion CHOP pathway of endoplasmic reticulum stress is involved in renal damage caused by hyperlipidemia.

Effect of hyperlipidemia and influence of simvastatin on endoplasmic reticulum stress in rat kidney / 中国病理生理杂志

AIM: To investigate the role of endoplasmic reticulum stress in renal injury caused by hyperlipidemia and the influence effect of simvastatin. METHODS: Thirty male Wistar rats were randomly divided into three groups: rats in control group (n=10) were fed with normal diet; rats in high fat group (n=10) were fed with high fat diet; animals in simvastatin+high fat group (n=10) were fed with high fat diet and were received simvastatin 10 mg·kg~(-1)·d~(-1) by gastric irrigation. After 18 weeks, the quantitative urine protein in 24 h, the serum cholesterol and triglycerides levels were tested. The pathological changes of renal tissue were observed under optic microscope. The expressions of GRP78 and p-JNK in renal tissues were examined by immunohistochemistry. The apoptotic cells in the kidney were detected by TUNEL staining. The mRNA expressions of GRP78 and CHOP were examined by RT-PCR. RESULTS: The quantitative urine protein in 24 h, the serum lipid, the expressions of GRP78 and p-JNK proteins, the mRNA expressions of GRP78 and CHOP as well as the apoptotic cells in renal tissues were increased in high fat group (P<0.01).The quantitative urine protein in 24 h, the serum lipid, the expression of GRP78 and p-JNK proteins, the mRNA expressions of GRP78 and CHOP as well as the apoptotic cells in renal tissues were remarkably reduced in simvastatin+high fat group than those in high fat group (P<0.05). CONCLUSION: The endoplasmic reticulum stress is engaged in the renal injury caused by hyperlipidemia. The simvastatin play a role in renal protection by inhibiting the endoplasmic reticulum stress in the kidney.

Time and distribution characteristics of neuron apoptosis in rat after ischemic insult / 第三军医大学学报

Objective To investigate the time and distribution characteristics of neuron apoptosis occurred after global cerebral ischemia in rats,so as to provide some helpful experimental basis both for the early interference of injured neuron and the prevention and treatment of brain secondary damage.Methods In the global cerebral ischemia model with modified 4-vessel occlusion method,the amount and distribution of apoptotic neuron in hippocampus and cortex after ischemia and reperfusion were detected with TTC staining,TUNEL labeling and AO/EB fluorescence examination techniques.Intracellular free calcium concentration was measured with Fura-2 AM fluorescent indicator.Results TUNEL showed that apoptotic neurons scattered in hippocampus 3 h after reperfusion,then spread around to cortex.The amount of apoptotic cell reached to the peak at 24-48 h.The died cells appeared later and mainly dispersed around the apoptotic cells.There was a significant increase of the amount of died cells 72 h after ischemia and reperfusion.AO/EB fluorescence examination showed that the total amount of the apoptotic neurons in hippocampus,frontal cortex and parietal cortex reached their peak at 24 h and 72 h respectively,and they were significantly higher than that in control group(P

The regulation mechanism of shRNAspten to NR2B after neuron subjected to anoxia insult / 重庆医科大学学报

Objective:To investigate the regulation of PTEN with RNAi on NR2B after OGD.Methods:In the OGD model with anaerobic gas mixture and deoxygenated,glucose-free extracellular solution,shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively.The quantities of NR2B in the neuronal membrane were quantitatively measured with colorimetric assay,and the expression of NR2B mRNA were analyzed by RT-PCR.The neuronal activity was analyzed by AlamarBlue andintracellular free calcium concentration were measured with Fura-2 AM by laser confocal microscope(LCM).Results:shRNAspten-GFP plasmid can been expressed successfully in cultured neurons.The study demonstrated that there are numerous NR2B subunit expression on neuronal membrane by colorimetric assay(OPD)assay and RT-PCR.The expression and quantity of neuronal NR2B with shRNAspten-GFP treatment was significantly decrease after oxygen and glucose deprivation(OGD)and reperfusion(P

The mechanism of PTEN down-regulation on neuron intracellular free calcium and apoptosis / 重庆医科大学学报

Objective: To investigate the regulation of PTEN on apoptosis and the active roles in blocking Ca2+ influx and neuroprotection.Methods: In the OGD model with anaerobic gas mixture and deoxygenated,glucose-free extracellular solution,shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively.Intracellular free calcium concentration were measured with Fura-2 AM fluorescent indicator,the expression of PTEN were analyzed by immunoblotting(blot) and neuron apoptosis were detected by AO/EB and Flow Cytometry.Results: Apoptosis rate and calcium concentration after OGD was significantly higher than that of normal group(P

A proteomic approach to study cerebrospinal fluid from migraine patients / 解放军医学杂志

Objective To identify the biomarkers associated closely with migraine by proteomics analysis of cerebrospinal fluid (CSF).MethodsForty-five patients were involved in the present study and they were divided into the following two groups:25 with migraine (5 with aura and 20 without aura) were assigned to migraine group,and 20 with giddiness or peripheral neuropathy were assigned to control group.The patients' CSF was collected,and the protein of CSF was extracted by acetone precipitation.The two-dimensional electrophoresis (2D-PAGE) with immobilized pH gradient (IPG) was performed to display the differently expressed protein spots.These spots were then identified by two-dimensional liquid chromatography and tandem mass spectrometry (2D-LC/MS-MS),and semi-quantitatively analyzed by Western blotting.ResultsDifferences were found in the results of 2D-PAGE between migraine group and control group.Ten spots of 9 kinds of proteins were successfully identified,of which 7 spots were down regulated,including transthyretin (TTR),CBX6 53 kDa protein (CBX6),agrin precursor (AGRN),FAM3C precursor (FAM3C),neuronal pentraxin receptor (NPR),dermcidin-isform2 (DCD) and albumin (ALB);whereas junction plakoglobin (JUP) and H2A histone family,member J (H2AFJ) were up-regulated.The results of Western blotting revealed that the net protein retention (NPR) in migraineurs group (0.3351?0.0275) was obviously decreased compared with that of control group (0.8854?0.0957,P