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Although human antibodies elicited by severe acute respiratory distress syndrome coronavirus-2 (SARS-CoV-2) nucleocapsid (N) protein are profoundly boosted upon infection, little is known about the function of N-directed antibodies. Herein, we isolated and profiled a panel of 32 N protein-specific monoclonal antibodies (mAb) from a quick recovery coronavirus disease-19 (COVID-19) convalescent, who had dominant antibody responses to SARS-CoV-2 N protein rather than to Spike protein. The complex structure of N protein RNA binding domain with the highest binding affinity mAb nCoV396 reveals the epitopes and antigens allosteric changes. Functionally, a virus-free complement hyper-activation analysis demonstrates that nCoV396 specifically compromises N protein-induced complement hyper-activation, a risk factor for morbidity and mortality in COVID-19, thus paving the way for functional anti-N mAbs identification. One Sentence SummaryB cell profiling, structural determination, and protease activity assays identify a functional antibody to N protein.
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The outbreak of coronavirus disease (COVID-19) in China caused by SARS-CoV-2 virus continually lead to worldwide human infections and deaths. It is currently no specific viral protein targeted therapeutics yet. Viral nucleocapsid protein is a potential antiviral drug target, serving multiple critical functions during the viral life cycle. However, the structural information of SARS-CoV-2 nucleocapsid protein is yet to be clear. Herein, we have determined the 2.7 [A] crystal structure of the N-terminal RNA binding domain of SARS-CoV-2 nucleocapsid protein. Although overall structure is similar with other reported coronavirus nucleocapsid protein N-terminal domain, the surface electrostatic potential characteristics between them are distinct. Further comparison with mild virus type HCoV-OC43 equivalent domain demonstrates a unique potential RNA binding pocket alongside the {beta}-sheet core. Complemented by in vitro binding studies, our data provide several atomic resolution features of SARS-CoV-2 nucleocapsid protein N-terminal domain, guiding the design of novel antiviral agents specific targeting to SARS-CoV-2.
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Objective:To investigate the application value of transcranial Doppler (TCD) in the prognosis assessment of nerve function in patients with acute cerebral infarction (ACI) after intracranial mechanical thrombectomy.Methods:A retrospective analysis was conducted. The clinical data of 43 patients with acute anterior circulation cerebral infarction who received intra-arterial mechanical thrombotomy for recanalization admitted to Taizhou Central Hospital from January 2018 to December 2019 were analyzed. The modified Rankin scale (mRS) score of patients were followed up by telephone at 3 months after surgery to evaluate the prognosis of neurologic outcome. Patients with mRS score 0-2 were enrolled in the good prognosis group, while those with a score of 3-6 were enrolled in the poor prognosis group. The gender, age, past history, underlying diseases, occluded arteries, atherosclerotic stenosis and bridging treatment, time from onset to reperfusion, blood flow dynamics under TCD at 1 day after thrombectomy, and National Institutes of Health stroke scale (NIHSS) scores before and 1, 7, and 14 days after thrombectomy were compared between the two groups. Multivariate Logistic regression analysis was used to screen the prognostic factors of nerve function at 3 months after mechanical thrombectomy in patients with ACI. The receiver operating characteristic (ROC) curve was drawn to evaluate the prognostic value for neurological function assessed by TCD.Results:Forty-three patients were enrolled in the final analysis, with 23 patients in the good prognosis group and 20 in the poor prognosis group. The recanalization was successfully achieved in both groups without complications. However, the hemodynamics of intracranial arteries evaluated by TCD 1 day after operation in both groups still showed partial or complete occlusion, and the hemodynamics of patients in the poor prognosis group was worse than that in the good prognosis group (poor blood flow: 40.0% vs. 0%, inadequate blood flow: 30.0% vs. 17.4%, good blood flow: 30.0% vs. 82.6%), and the differences were statistically significant (all P < 0.01). Before thrombotomy, there was no significant difference in NIHSS score between the two groups. After thrombotomy, the NIHSS score of the two groups gradually decreased with the extension of time, but the NIHSS score at 14 days after operation of the poor prognosis group was still significantly higher than that of the good prognosis group (10.55±2.93 vs. 4.65±1.70, P < 0.01). Univariate analysis showed that compared with the good prognosis group, the proportion of patients with diabetes and arteriosclerosis stenosis in the poor prognosis group were significantly increased (30.0% vs. 4.3%, 45.0% vs. 17.4%, both P < 0.05), and the time from onset to reperfusion was prolonged (minutes: 385.9±96.2 vs. 294.5±95.1, P < 0.01). Multivariable Logistic regression analysis showed that the therosclerosis stenosis [odds ratio ( OR) = 9.334, 95% confidence interval (95% CI) was 1.092-79.775, P = 0.041] and the reperfusion time ( OR = 1.016, 95% CI was 1.006-1.027, P = 0.002) were associated with prognosis of nerve function at 3 months after mechanical thrombectomy in patients with ACI. ROC curve analysis suggested that the evaluation of intracranial hemodynamics by TCD might be able to predict the prognosis of neurological function in patients with ACI after 3 months of intracranial mechanical thrombectomy, the area under ROC curve (AUC) was 0.768 (95% CI was 0.620-0.917), the sensitivity was 65.0%, the specificity was 87.0%, the positive predictive value was 82.6%, and the negative predictive value was 70.0%. Conclusion:The evaluation of intracranial hemodynamics assessed by TCD is helpful in early judging the prognosis of neurological function in patients with ACI after intracranial mechanical thrombectomy.
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The outbreak of coronavirus disease (COVID-19) caused by SARS-CoV-2 virus continually lead to worldwide human infections and deaths. Currently, there is no specific viral protein-targeted therapeutics. Viral nucleocapsid protein is a potential antiviral drug target, serving multiple critical functions during the viral life cycle. However, the structural information of SARS-CoV-2 nucleocapsid protein remains unclear. Herein, we have determined the 2.7 Å crystal structure of the N-terminal RNA binding domain of SARS-CoV-2 nucleocapsid protein. Although the overall structure is similar as other reported coronavirus nucleocapsid protein N-terminal domain, the surface electrostatic potential characteristics between them are distinct. Further comparison with mild virus type HCoV-OC43 equivalent domain demonstrates a unique potential RNA binding pocket alongside the -sheet core. Complemented by binding studies, our data provide several atomic resolution features of SARS-CoV-2 nucleocapsid protein N-terminal domain, guiding the design of novel antiviral agents specific targeting to SARS-CoV-2.
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Objective To investigate the effect of hepatitis B virus (HBV) X gene integration on expression of zinc finger protein ZBTB20 in chronic hepatitis B (CHB) patients complicated with hepatocellular carcinoma (HCC).Methods Eighteen CHB patients complicated with HCC who underwent surgical treatment in Taizhou Enze Medical Center Enze Hospital and Taizhou Central Hospital from July 2015 to June 2017 were enrolled.Samples of carcinoma tissue,para-carcinoma tissue and corresponding normal liver tissue were collected from each case.DNA was extracted from three kinds of tissue samples.HBV-Alu-polymerase chain reaction (PCR) was used to amplify the integrated HBVX fragments and their bilateral flanking sequences in human genomic DNA.The integrated HBV fragments were determined by PCR products sequencing.Protein was extracted from three kinds of tissue samples.The level of expression of ZBTB20 was detected by protein imprinting.Statistical analysis was performed using t test,analysis of variance,and x2 test.Results Among the 18 CHB patients complicated with HCC,integration of HBVX gene was found in 13 carcinoma tissue samples,16 para-carcinoma tissue samples and 9 corresponding normal liver tissue samples.The difference was statistically significant (x2 =6.353,P =0.037).Of the 18 patients,the protein expressions of ZBTB20 in carcinoma tissue,para-carcinoma tissue and corresponding normal tissue were (50.14 ± 11.25)%,(40.71±7.17) % and (39.06 ± 5.17) %,respectively,which was statistically different (F =9.420,P < 0.01).HBVX gene integration was detected at ZBTB20 locus in five patients.The expression levels of ZBTB20 in patients with HBVX gene integration at this locus in carcinoma tissue,para-carcinoma tissue and corresponding,normal liver tissue were all significantly lower than those in patients without HBVX gene integration (carcinoma tissue (37.37±10.30)% vs (55.06 ± 7.06)%,para-carcinoma tissue (32.06 ± 2.61)% vs (44.04 ± 5.24)%,corresponding normal tissue (34.66 ±5.59)% vs (40.76 ±4.04)%,t--4.205,4.821 and 2.589,respectively,all P <0.05).Conclusions Incidence of HBVX integration in para-carcinoma tissue is higher than that in carcinoma tissue in CHB patients complicated with HCC.The expression level of ZBTB20 in carcinoma tissue is higher than that in para-carcinoma tissue.Integration of HBVX gene at ZBTB20 locus may decreases the expression of ZBTB20.
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Objective@#To investigate the effect of hepatitis B virus (HBV) X gene integration on expression of zinc finger protein ZBTB20 in chronic hepatitis B (CHB) patients complicated with hepatocellular carcinoma (HCC).@*Methods@#Eighteen CHB patients complicated with HCC who underwent surgical treatment in Taizhou Enze Medical Center Enze Hospital and Taizhou Central Hospital from July 2015 to June 2017 were enrolled. Samples of carcinoma tissue, para-carcinoma tissue and corresponding normal liver tissue were collected from each case. DNA was extracted from three kinds of tissue samples. HBV-Alu-polymerase chain reaction (PCR) was used to amplify the integrated HBVX fragments and their bilateral flanking sequences in human genomic DNA. The integrated HBV fragments were determined by PCR products sequencing. Protein was extracted from three kinds of tissue samples.The level of expression of ZBTB20 was detected by protein imprinting. Statistical analysis was performed using t test, analysis of variance, and χ2 test.@*Results@#Among the 18 CHB patients complicated with HCC, integration of HBVX gene was found in 13 carcinoma tissue samples, 16 para-carcinoma tissue samples and 9 corresponding normal liver tissue samples. The difference was statistically significant (χ2=6.353, P=0.037). Of the 18 patients, the protein expressions of ZBTB20 in carcinoma tissue, para-carcinoma tissue and corresponding normal tissue were (50.14±11.25)%, (40.71±7.17)% and (39.06±5.17)%, respectively, which was statistically different (F=9.420, P<0.01). HBVX gene integration was detected at ZBTB20 locus in five patients. The expression levels of ZBTB20 in patients with HBVX gene integration at this locus in carcinoma tissue, para-carcinoma tissue and corresponding, normal liver tissue were all significantly lower than those in patients without HBVX gene integration (carcinoma tissue (37.37±10.30)% vs (55.06±7.06)%, para-carcinoma tissue (32.06±2.61)% vs (44.04±5.24)%, corresponding normal tissue (34.66±5.59)% vs (40.76±4.04)%, t=4.205, 4.821 and 2.589, respectively, all P<0.05).@*Conclusions@#Incidence of HBVX integration in para-carcinoma tissue is higher than that in carcinoma tissue in CHB patients complicated with HCC.The expression level of ZBTB20 in carcinoma tissue is higher than that in para-carcinoma tissue. Integration of HBVX gene at ZBTB20 locus may decreases the expression of ZBTB20.
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Objective To explore the effect of lean management on cost control of single disease in patients with acute cerebral infarction (ACI) in stroke center. Methods A retrospective study was conducted. The patients with ACI who underwent intravenous thrombolysis in the stroke center of Taizhou Central Hospital in Zhejiang Province were enrolled. Thirty patients adopted traditional management procedures from July 2016 to September 2017 were enrolled in the control group, and 32 patients received lean management from October 2017 to December 2018 were enrolled in the lean group. The patients in the control group were treated with traditional intravenous thrombolysis, and the patients were sent to the neurology ward for intravenous thrombolysis. The patients in the lean group applied lean management value stream to optimize process management, the lean management team of the stroke center was established, and the green channel for stroke treatment was established to eliminate the waiting time as far as possible. The location of thrombolysis was changed from neurology ward to the neurological intensive care unit (NICU) in emergency department. The patients in the two groups were compared in terms of intravenous thrombolytic door-to-needle time (DNT), admission time to the neurologist's visit time (T1), CT examination time to neurology ward or NICU admission time (T2), neurology ward/NICU visit time to medication time (T3), and the proportion of patients with DNT controlled within 40 minutes, recovery of neurological impairment 7 days after thrombolysis [national institutes of health stroke scale (NIHSS) score], activity of daily living assessment (Barthel index), length of hospital stay, cost of hospital stay and patient satisfaction. At the same time, the main process quality and the implementation rate of easily missed indexes of cerebral infarction single disease were recorded. Results Compared with the control group, DNT, T1 and T2 in the lean group were significantly shortened [DNT (minutes): 39.56±11.12 vs. 63.03±19.63, T1 (minutes): 16.23±6.79 vs. 33.48±12.63, T2 (minutes): 13.45±3.84 vs. 17.47±5.56, all P < 0.01], T3 was slightly shortened (minutes: 9.88±1.95 vs. 10.95±2.69, P > 0.05), and the proportion of DNT control within 40 minutes was significantly increased [75.0% (24/32) vs. 16.7% (5/30), P < 0.01], the 7-day NIHSS score was decreased significantly (8.66±4.12 vs. 13.00±5.63, P < 0.01), 7-day Barthel index was increased significantly (71.6±16.7 vs. 54.7±17.1, P < 0.01), the length of hospital stay was significantly shortened (days: 9.69±4.06 vs. 12.47±3.83, P < 0.01), the hospital costs were significantly reduced (Yuan: 16 338±5 481 vs. 19 470±5 495, P < 0.05), the satisfaction of patients was improved significantly [(91.38±2.69)% vs. (86.53±2.78)%, P < 0.01]. In terms of the implementation rate of quality indicators such as pre-application evaluation of thrombolytic drugs, evaluation of dysphagia, and evaluation of vascular function, health education of ACI, rehabilitation evaluation and implementation within 24 hours, etc., the lean group was significantly improved as compared with the control group [(87.5% (28/32) vs. 53.3% (16/30), 96.9% (31/32) vs. 73.3% (22/30), 78.1% (25/32) vs. 43.3% (13/30), 100.0% (32/32) vs. 76.7% (23/30), 75.0% (24/32) vs. 33.3% (10/30), all P < 0.05]. Conclusion Lean thinking can realize the standardization of stroke center process, effectively utilize medical resources, improve medical quality and reduce the cost of cerebral infarction single disease.
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Objective To investigate the effect of intermittent high glucose on oxygen-glucose deprivation/refurnish (OGD/R) neuronal survival.Methods The primary cultured hippocampal neurons of mice were sub-cultured when the cell fusion reached about 80%.Cells in logarithmic growth phase were placed in a hypoxic incubator (37 ℃,5% CO2,95% N2) to simulate cell hypoxia.The culture medium was replaced by glucose-free Hank equilibrium salt solution (HBSS) to simulate cell hypoglycemia.The normal glucose and oxygen control group was set up.Cell morphology was observed under inverted phase contrast microscope after 6 hours of hypoxia and hypoglycemia treatment,and cell viability was detected by CCK-8 cell proliferation assay kit,and then grouping experiment was carried out.The cells were randomly divided into four groups.The cells were cultured in different concentration glucose medium under normal oxygen,5% CO2 and 37 ℃ for 72 hours to prepare OGD/R model of cell ischemia/reperfusion.The low-glucose control group was cultured in medium containing 5.5 mmoFL glucose.The constant high-glucose group was cultured in medium containing 33.0 mmol/L glucose.The intermittent high-glucose group was cultured in medium containing 33.0 mmol/L glucose for 3 hours then in medium containing 5.5 mmol/L glucose for 2 hours alternately for 3 times during the day,and overnight in medium containing 33.0 mmol/L glucose at night.The hyperosmotic control group was made up of 5.5 mmol/L glucose medium and mannitol.The osmotic pressure was the same as that of the constant high-glucose group,and the effective glucose concentration was the same as that of the normal glucose and oxygen group,so as to eliminate the effect of osmotic pressure changes caused by the high-glucose medium on the results.Cell morphology was observed under inverted phase contrast microscope after 72 hours of cell culture in each group.Cell viability was measured by CCK-8 kit,and apoptotic rate was measured by flow cytometry.Results The inverted phase contrast microscope showed that the cells in the normal glucose and oxygen control group were plump and refractive,and had obvious nucleus,clear processes and high cell activity.After 6 hours of hypoxia and hypoglycemia treatment,the cells were shrunk,refractive index was poor,the nucleus was unclear,the processes were not clear,and the cell activity was significantly lower than that of normal glucose and oxygen control group (A value:0.34±0.06 vs.1.09±0.06,P < 0.01),which indicated that the model of oxygen-glucose deprivation (OGD) was successfully prepared.After 72 hours of culture with different concentrations of glucose,the cells in the low-glucose control group were shrunk,the cell membrane was incomplete,the nucleus was unclear,and number of necrotic cells were more.In the constant high-glucose group,the refractive index of cells was poor,a large number of cells floated,and the nucleus was not obvious.In the intermittent high-glucose group,the cell morphology was normal,the refractive rate of cells was decreased slightly,and the necrotic cells were less.In the hypertonic control group,the cell status was close to that in the constant high-glucose group.Compared with the low-glucose control group or constant high-glucose group,the cell viability in the intermittent high-glucose group was significantly increased (A value:2.04±0.15 vs.0.64±0.18,1.16±0.16,both P < 0.01),the apoptotic rate was significantly decreased [(59.60 ± 2.55)% vs.(78.15 ± 15.77)%,(95.60± 0.14)%,both P < 0.05].There was no significant difference in cell activity or apoptotic rate between the hypertonic control group and the constant high-glucose group [cell activity (A value):1.07 ± 0.07 vs.1.16 ± 0.16,apoptotic rate:(87.80 ± 4.53)% vs.(95.60 ± 0.14)%,both P > 0.05].Conclusion Intermittent high glucose within a certain range had protective effect on OGD/R neuronal survival.
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Objective To explore the effect of omni-directional health education on self-management level and blood glucose control in patients with type 2 diabetes mellitus. Methods A total of 100 diabetic patients admitted to the department of neurology of Taizhou Central Hospital from January to December 2016 were randomly divided into an observation group and a control group, 50 cases in each group. All patients were given conventional medication according to their systemic conditions, the control group used conventional nursing care, while the observation group was supplemented with omni-directional health education (including diet control therapy, exercise therapy, blood glucose control, hypoglycemic drug use and observation, foot care, blood glucose self monitoring) on the basis of routine nursing;the course of treatment was 2 weeks in the two groups. The differences in self-management level and the effect of blood glucose control were compared between the two groups. Results On the return visits, the self-management scores were significantly higher in both groups than those before education (the control group was 35.2±9.2 vs. 30.1±5.2, the observation group was 78.6±11.5 vs. 29.8±5.4, all P < 0.05), fasting blood glucose [FBG (mmol/L): the control group was 8.8±2.5 vs. 14.8±2.9, the observation group was 6.1±1.6 vs. 15.1±2.7, both P < 0.05) and 2 hours postprandial blood glucose [2 h PBG (mmol/L): the control group was 12.5±3.4 vs. 22.5±3.8, the observation group was 9.1±2.2 vs. 21.9±3.6) were obviously lower than those before education (all P < 0.05), the degrees of improvement of the observation group were more significant than those of the control group [self management score: 78.6±11.5 vs. 35.2±9.2, FBG (mmol/L): 6.1±1.6 vs. 8.8±2.5, 2 hours PBG (mmol/L): 9.1±2.2 vs. 12.5±3.4, all P < 0.05]. Conclusion Omni-directional health education can improve the self-management level of diabetic patients, and the patient's blood glucose control is relatively better.
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Objective To compare the value of ultrasound International Ovarian of Tumor Analysis (IOTA) simple rules and gynecologic imaging reporting and data system (GI-RADS) ultrasonographie stratification in the diagnosis of adnexal masses.Methods A total of 463 patients with adnexal masses were classified according to IOTA simple rules by senior doctor.The benign and malignant masses were enrolled in the study.Then the masses were judged according to GI-RADS classification and IOTA simple rules by senior and junior doctors.And junior doctorn in diagnosis of adnexal masses with IOTA was group A1,senior doctor in diagnosis of adnexal masses with IOTA was group B1,junior doctor in diagnosis of adnexal masses with GI-RADS was A2,senior doctor in diagnosis of adnexal masses with GI-RADS was B2.The efficacy of the two methods were compared.Results Among the 463 cases,there were 411 cases (411/463,88.77%) of benign mass and malignant mass by IOTA and 52 cases (52/463,11.23 %) of uncertain mass.The specificity,positive predictive value,diagnostic accuracy had statistical differences between groups A2 and B2,A1 and A2 (all P<0.05),and sensitivity and negative predictive value had no statistical differences (all P>0.05).Diagnostic efficacy index had no statistial signifi cance between group A1 and B1,B1 and B2 (all P>0.05).Conclusion The diagnostic efficacy of IOTA simple rules and GI-RADS is high and similar in diagnosis of benign and malignant adnexal masse.IOTA simple rules do not depend on experience,but they are not suitable for the diagnosis of all adnexal masses.
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Objective To evaluate the value of fetal dual Doppler ultrasonography in the diagnosis of fetal arrhythmias.Methods 77 53 1 fetuses were examined by echocardiography.If fetal arrhythmias were found,information of pulsed wave Doppler signals in hepatic vein(HV) and descending aorta(DAo) using dual Doppler were collected simultaneously.Results On dual Doppler examination,13 1 of the fetuses showed signals of cardiac dysrhythmias,including 60 fetuses with irregular fetal heart rhythm but having normal heart rate,45 with tachyarrhythmias and 26 with bradyarrhythmias.Sixteen and 2 with tachy- or bradyarrhythmias fetuses complicated fetal malformation or severely maternal systemic diseases.Eighteen and 3 with tachy- or bradyarrhythmias fetuses revealed intrauterine death or intermediate pregnancy induction.The three main kinds of fetal arrhythmias had statistical differences in complicating with fetal malformation,maternal diseases,intrauterine fetal demise or intermediate pregnancy induction(all P <0.05).There was a significant difference in the accuracy of diagnosing rhythm abnormalities in the fetus between conventional ultrasonography and dual Doppler ultrasonography(k = 0.562,P < 0.05).Sixty cases of irregular fetal heart rhythm,42 cases of fetal tachyarrhythmias,and 8 cases of fetal bradyarrhythmias were changed to normal rhythm at the third-pregnancy or prenatal period through appropriate treatment.All surviving fetuses were examined by echocardiography within 3 days,without any combination of congenital heart disease,and 1 months after the delivery,the ECG were shown as sinus rhythm.Conclusions Dual Doppler ultrasonography is a precise and rapid method of assessing various types of fetal arrhythmias.Different types of fetal arrhythmias have different fetal prognosis.
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Objective To observe the effects of hirudo capsules on degree of platelet activation and blood rheology in patients with cerebral arteriosclerosis. Methods 122 patients with cerebral arteriosclerosis were randomly divided into two groups by the random number table:observation group(61 cases)and control group (61 cases). The two groups received conventional treatment such as antihypertensive and hypoglycemic drugs,and 100 mg of aspirin once a day. In the observation group,additionally hirudo capsules 0.75 g,3 times a day were given. The therapeutic course in both groups was 2 months. Prior to the start of treatment and on the next day after the end of treatment,traditional Chinese medicine(TCM)clinical syndrome integral,platelet membrane glycoproteins caspase 1 (PAC-1),platelet adhesion molecule P selection(CD62P)positive rate,and blood rheology indexes were detected,in addition to the adverse reactions . Results Before treatment,there were no statistical significant differences between the two groups in terms of TCM clinical syndrome integral such as headache,dizziness,head fullness,multiple somnolence amnesia,lip dark purple,sublingual veins purple in color,PAC-1,CD62P positive rate,hemorheology indexes(all P>0.05). After treatment,TCM clinical syndrome integral PAC-1,CD62P,low shear viscosity,high shear viscosity,plasma viscosity and the hematocrit of two groups were significantly lower than those before treatment, the improvement in observation group being more remarkable〔headache:0.75±0.69 vs. 1.23±0.92,dizziness:0.96±0.78 vs. 1.54±1.24,head fullness:0.65±0.59 vs. 1.48±1.17,multiple somnolence amnesia:0.77±0.72 vs. 1.69±1.14,lip dark purple:0.83±0.81 vs. 1.32±0.79,sublingual veins purple:0.84±0.67 vs. 1.20±0.74, PAC-1:(10.31±4.57)%vs.(15.13±6.27)%,CD62P:(11.39±5.24)%vs.(16.40±7.91)%,low shear viscosity (mPa?s):7.52±3.37 vs. 8.97±3.50,high shear viscosity(mPa?s):4.35±1.12 vs. 5.58±1.19,plasma viscosity (mPa?s):1.54±0.35 vs. 1.88±0.42,hematocrit(HCT):0.43±0.09 vs. 0.49±0.10〕. In the patients' follow-up, there were no obvious side effects of hirudo capsules. Conclusion Hirudo capsule is a safe and effective drug for treatment of patients with cerebral arteriosclerosis as it is helpful in reducing platelet activation degree and improving the blood rheology indexes.
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Objective To study the expression and the mechanism of miR-155in the villi of patients with unexplained recurrent spontaneous abortion (URSA).Methods The expression of miR-155 in the villi of 36 cases with URSA (URSA group) and 25 women with normal early pregnancy (control group) were detected by stem-loop real-time reverse transcription (RT) qPCR.Expression of hypoxia inducible factor-1 (HIF-1α),vascular endothelial cell growth factor(VEGF) and micro lymphatic vessel density (MVD) in the villi of were measured by immnohistochemical staining among two groups.Results (1) miR-155 expression:the mean miR-155 expression were 1.456 (0.489,2.459) in URSA group and 2.833 (1.740,3.794) in control group,which reached statistical difference (P <0.05).The mean expression of miR-155 of 1.683 (0.902,2.459) in URSA group with abortion times (≤ 3) was significantly higher than 1.229 (0.489,1.719) in URSA group with more than 4 times abortion (P < 0.05).(2) Indexes:the expression of HIF-1α,VEGF and MVD value were 121 ± 12,134 ± 12,36 ± 6 in URSA group and 99 ± 10,109 ± 10,28 ±4 in control group,which reached statistical difference(P < 0.01).The expression of HIF-1α,VEGF and MVD value of 119 ± 12,134 ± 12,35 ± 5 in URSA group with less than 3 times abortion was significantly lower than 128 ± 12,138 ± 12,43 ± 6 in URSA group with more than 4 times abortion (P < 0.01).Conclusions The expression of miR-155 and HIF-1α is topically stimulated by oxygen signal.HIF-1α adjusts the transcription and translation of VEGF,which together involved in placental trophoblast invasion and placental angiogenesis.The low expression of miR-155 could interfere with expression of HIF-1α and VEGF,which might be involved in villous vascular dysplasia in URSA.
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Objective To evaluate the value of dual Doppler ultrasonography in the diagnosis of fetal atrial extrasystoles.Methods Seven thousands and thirteen fetuses were examined by fetal echocardiography.If fetal arrhythmias were found,information of pulsed wave Doppler signals in hepatic vein (HV) and descending aorta (DAo) using dual Doppler were colleted simultaneously.The detected times were compared between using general Doppler and dual Doppler in 20 cases.Results On dual Doppler examination,28 of the fetuses showed signals of atrial extrasystoles,14 fetuses atrial extrasystoles conducted to ventricle,8 fetuses atrial extrasystoles non-conducted to ventricle,6 fetuses with part of the extrasystoles conducted to ventricle and part of the extrasystoles non-conducted to ventricle.No case had congenital heart disease.Twenty-six fetuses with atrial extrasystole were converted to normal rhythm within the third trimester of pregnancy,and the others' were converted to normal rhythm during the first 3 days postnatally.The detected time of dual Doppler was shorter than that of general Doppler(P <0.05).Conclusions Dual Doppler ultrasonography of simultaneous recordings in HV and DAo is an easy,rapid method of assessing fetal atrial extrasystoles,which should allow precise diagnosis of atrial extrasystole.
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Experimental course of pathology is a very important part of pathology teaching.Motivation of students was promoted through improving the professional knowledge of teachers,using teacher-student interacted teaching methods,increasing network courscs,using multimedia in teaching and adding extracurricular practice in our university.In the meantime,it was successful and fruitful to use diversified evaluation systems to assess the comprehensive experimental capacity of students,which can provide experiences in reform of pathological experimental teaching.
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Objective To investigate the distribution of eosinphililic neurons ( ENs),reactive astrocytes ( RAs),and infarction after transient cerebal ischemia,and the time profile of pathomorphological changes.Methods Unilateral forebrain ischemia was induced in Mongolian gerbils by two 10 minutes unilateral common carotid artery occlusions with a 5 hours interval.Laser Doppler flowmetry was used to detect intra-ischemic anterior cortex blood flow.Animals were sacrificed at 24 hours,4 days,2 weeks,4 weeks,16 weeks and the brain were prepared for pathomorphological assay.Results Intra-ischemic laser Doppler flowmetry show significant ischemia during carotid artery occlusion:22.1% ± 9.5%,26.3% ± 4.9%,37.5% ± 3.5%,F =67.219,P < 0.01 ; the decrease was significantly greater in the anterior cortex.ENs appeared in middle and deep layers at 24 hours postischemia,and ENs area extend to all layers of cortex by 4 days.Large areas of high EN density ( ≥80/mm2) evolved to infarcts between 4 days and 4 weeks.Posterior cortex evolved to low EN area ( < 80/mm2) without transformation into infarcts.RAs were consistently distributed in areas with ENs,and RA areas with high EN density were largely transformed into infarcts between 4 days and 4 weeks. Delayed astrocytic death took place in the RA areas with high EN density.Conclusion Density of ENs is an important indicator of delayed astrocytic death and infarction in postischemic tissue.
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Sixty patients who had hemodialysis over half year were randomly divided into 3 groups with 20 cases in each group:group A received conventional hemodialysis,group B received hemodialysis with oral administration of compound danshen (salvia miltiorrhiza bunge) tablets and vitamin E,group C had the same treatment as group B but the dialyzer was pretreated with solution containing 5 g vitamin C ; 20 healthy adults served as normal controls. Serum levels of advanced oxidation protein products (AOPP) and superoxide dismutase (SOD) were measured in all groups.The levels of AOPP in groups A,B,C were higher and SOD levels were lower than those of normal controls ( all P < 0.01 ).The levels of AOPP in groups B,C were lower and SOD were higher than those in group A at the same time points (P <0.01 ).The levels of AOPP in group C were lower and SOD were higher than those in group B at the same time points (P <0.0l ).The results suggest that pretreatment of the dialyzer with high dose vitamin C plus vitamin E and compound danshen tablets can alleviate oxidative stress in patients with maintaining hemodialysis.
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Objective To investigate the relationship between CD158j expression and phosphorylated ERK (p-ERK) in CD4+ CD28null T cells in cerebral infarction (CI) patients- with carotid atherosclerosis and its effects on carotid atherosclerotic plaque stability. Methods Percentage of peripheral CD4+ CD28null and the expression of CD158j and perform on CD4+ CD28null cells was analyzed with flow cytometry in 106 CI patients with carotid atherosclerosis, 33 CI patients with normal carotid arteries and in 50 normal controls, respectively; p-ERK expression was assayed with flow cytometry in 36 CI patients with unstable plaque, and serum IFN-γ was detected with ELISA. The intima-media thickness (IMT) of bilateral carotid arteries in all subjects was confirmed by the colour Doppler ultrasonograph imagingResults Percentage of the CD4+ CD28null T cells, expression of CD158j and perform on CD4+ CD28null T cells and the serum IFN-γ levels was dramatically higher in CI patients than that in normal controls, respectively (all P <0.01), which was decreased in an order of CI patients with patients with unstable plaque, stable plaque, carotid artery IMT and with normal carotid artery. A strong positive correlation was observed between the CD158j expression and degree of p-ERK in CI patients with unstable plaque (P < 0. 01). Conclusion CD4+ CD28null T cells were significantly increased in CI patients with carotid atherosclerosis. CD158j might up-regulate p-ERK expression and induce the proliferation of the CD4+ CD28nullT cells; consequently, higher cytokine production such as IFN-γ produced by CD4+ CD28null T cells may cause the formation of unstable plaque.
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Objective To study the hemodynamic characteristics of ductus venosus,and its relationship to inflammatory cytokines in ovine fetus with systemic inflammatory response syndrome.Methods Ten near-term fetal sheep radomly divided into two groups.Five of them were in experimental group,and five of them were in control group.All of the animals underwent abdominal cordocentesis guided by ultrasound at the term of 120-130 days,with a LPS injection at a dosage of 10 μg/kg of fetal weight in experimental group,and the same quantity of 0.9% NaCl solution in control.Doppler echocardiography were performed to determine hemodynamics changes of ductus venosus at 0.5 hour before the LPS injection,and at 1 hour,3 hours,6 hours after LPS injection.Meanwhile,fetal umbilical vein blood was sampled for ELISA essay of serum TNF-α and IL-6 at each of the above time points.Results In experimental group,the ductus venosus waveform PI values(DVPI),maximum velocity during cardiac ventricle systole(S)and during cardiac ventricle diastole(D),ductus venosus index(DVI),ductus venosus blood flow(DVQ),the ratio of S and ductus venous maximum velocity during atrial contracton(S/A),(S-A)/D increased with the time after LPS injection,and still significantly increased as compared with the control animals(all P<0.05).A decreased with the time after LPS injection,and still significantly changed as compared with the controls(P<0.05).And the ductus venosus diameter(DVD)had not obvious change(P>0.05).There were significantly positive correlations between DVPI and TNF-α,IL-6(all P<0.05),negative corelations between A and TNF-α,IL-6(all P<0.05),and positive correlations between S wave,DVI,DVQ,D wave,S/A,(S-A)/D and IL-6(all P<0.05),but not obvious relationships between S wave,DVI,DVQ,D wave,S/A,(S-A)/D and TNF-a(all P>0.05).Condusions DVPI and A might be easy and useful quantitative parameters in the evaluation of fetal systemic inflammatory respome syndrome with LPS injection.
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OBJECTIVE: To study the effectiveness of the pulsatility index for veins of ductus venosus (DV-PIV) and the Tei index in a prospective assessment of fetal hypoxic-ischemic brain damage in a near-term ovine fetus model with intermittent umbilical cord occlusion (UCO). METHODS: Twelve fetal sheep were studied with umbilical cord occlusion performed in the experimental group animals by complete inflation of an occluder cuff for 90 s, every 30 min for approximately 2.5 h. Fetal arterial blood was sampled at 5 min before the first umbilical cord occlusions, approximately 60 s of the first umbilical cord occlusions, and 3 min after each occlusion for blood gas, pH, neuron-specific enolase (NSE) and S100B. Doppler measurements and Doppler echocardiographic examinations were performed 5 min before the first umbilical cord occlusions and 3 min after each successive occlusion. RESULTS: In experimental group animals, UCO caused a large decline in arterial PaO(2) (to approximately 7.70 mmHg, p < 0.01), a modest decline in pH (to approximately 7.24, p < 0.01), and a modest rise in PaCO(2) (to approximately 53.31 mmHg, p < 0.01), with a return more or less to baseline after occluder release. and there was significant change as compared with the control animals (all p < 0.01) with cumulative changes in responses to repetitive cord occlusions. The DV-PIV waveforms, right ventricle (RV) and LV Tei indices, the serum levels of NSE and S100B increased with cord occlusions (all p < 0.05), and were significantly higher than the control animals (all p < 0.05) with a cumulative changes in responses to repetitive cord occlusions. RV and LV Tei indices were significantly correlated with PaO(2) (r = - 0.684, p < 0.01 and r = - 0.725, p < 0.01), PaCO(2) (r = 0.682, p < 0.01 and r = 0.780, p < 0.01), pH (r = - 0.538, p < 0.01 and r = - 0.681, p < 0.01), NSE (r = 0.653, p < 0.01 and r = 0.687, p < 0.01), and S100B (r = 0.606, p < 0.01 and r = 0.640, p < 0.01). Significant but weaker correlations were also present between DV-PIV and the parameters considered. CONCLUSION: Umbilical cord occlusion during the latter part of the pregnancy, enough to cause significant hypoxemia and acidosis, results in a significant increase of DV-PIV, RV and LV Tei indices, and the serum levels of NSE and S100B. There was a strong correlation between the RV and LV Tei indices and blood gas, pH, and NSE, S100B with hypoxia. Therefore, the Tei index might be an easy and useful quantitative parameter for assessing fetal hypoxic ischemia.