Objective:To evaluate the performance of the automated digital cell morphology instrument in detecting platelet (PLT) clumps.Methods:A total of 4271 blood samples whose PLT reached the reviewing rules of thrombocytopenia were selected from inpatients having blood analysis in Xijing Hospital from January 1 st to June 30 th, 2019, including 2 200 males and 2 071 females,with a median age of (35±7.03) years old. The smears for these cases were made, stained by Wright-Giemsa, and examined to capture PLT clumps by digital cell morphology system and manual microscope separately. The digital cell analysis system (hereinafter referred to as the instrument method) as an evaluation method and the microscope method as a reference method were used to calculate the positive rate of platelet clump detection and evaluate the comparison of two methods and bias assessments. The chi-square test was used to compare counting data rates. Results:Among 4, 271 samples reaching the reviewing rule of thrombocytopenia, 128 cases with platelet clumps were detected by manual microscope(initial) with a positive detection rate of 96.24%, and a total 133 of cases with PLT clumps were detected by microscope (initial+reconfirmation) with a positive detection rate of 100 %. Meanwhile, 129 cases with platelet clumps were detected by instrument method with a positive detection rate of 96.9%. There was no significant difference in terms of positive rate of PLT clumps detection between the instrumental method and the microscope method (initial) ( χ2 =0.115, P=0.73); the positive rate of clumps detection by the instrumental method was lower than microscope method (initial+reconfirmation), and the difference was statistically significant (χ 2 =4.061, P=0.04). For instrument method, the positive rate of PLT clumps detection by simultaneous observation of RBC analysis interface+PLT aggregation interface+WBC analysis interface was higher than only observation of PLT aggregation interface, and the difference was statistically significant (χ 2 =5.090, P=0.02). The average error of the deviation of PLT counting results before and after correction of the cases with PLT plumps missed by instrument method was significantly higher than microscope method (initial), and the difference was statistically significant (χ 2 =56.26, P<0.001). Conclusion:The automated digital cell morphology system has a good consistency with manual microscope(initial) in terms of the sensitivity of platelet clumps detection and can be used as a supplementary method for detecting platelet aggregation.

Objective:To investigate the influencing factors, periprocedural complications, and long-term outcomes of successful recanalization after endovascular treatment in patients with non-acute symptomatic internal carotid artery occlusion.Methods:Patients with non-acute internal carotid artery occlusion received endovascular treatment in the Nanjing Stroke Registration System between January 2010 and December 2021 were retrospectively enrolled. Clinical endpoint events were defined as successful vascular recanalization, periprocedural complications (symptomatic embolism and symptomatic intracranial hemorrhage), neurological function improvement, and recurrence of ipsilateral ischemic events. Multivariate logistic regression analysis was used to investigate the independent influencing factors of successful vascular recanalization. Cox proportional hazards regression analysis was used to investigate the correlation between endovascular treatment outcomes and neurological function improvement, as well as ipsilateral ischemic cerebrovascular events. Results:A total of 296 patients were included, of which 190 (64.2%) were successfully recanalized. Multivariate logistic regression analysis showed that symptoms manifest as ischemic stroke (odds ratio [ OR] 3.353, 95% confidence interval [ CI] 1.399-8.038; P=0.007), the time from the most recent symptom onset to endovascular therapy within 1 to 30 d ( OR 2.327, 95% CI 1.271-4.261; P=0.006), proximal conical residual cavity ( OR 2.853, 95% CI 1.242-6.552; P=0.013) and focal occlusion (C1-C2: OR 3.255, 95% CI 1.296-8.027, P=0.012; C6/C7: OR 5.079, 95% CI 1.334-19.334; P=0.017) were the independent influencing factors for successful vascular recanalization. Successful recanalization did not increase the risk of symptomatic intracranial hemorrhage within 7 d after procedure (3.2% vs. 0.9%; P=0.428). The median follow-up time after procedure was 38 months. Cox proportional hazards regression analysis showed that after adjusting for confounding factors, successful recanalization was significantly associated with postprocedural neurological improvement (hazard ratio 1.608, 95% CI 1.091-2.371; P=0.017), and significantly reduced the risk of recurrence of long-term ischemic events (hazard ratio 0.351, 95% CI 0.162-0.773; P=0.010). Conclusion:In patients with non-acute internal carotid artery occlusion, successful endovascular recanalization can effectively reduce the risk of long-term ischemic events without increasing the risk of symptomatic intracranial hemorrhage.

Objective:To investigate predictive factors for successful endovascular recanalization in patients with non-acute symptomatic internal carotid artery occlusion (SICAO), to develop a decision tree model using the Classification and Regression Tree (CART) algorithm, and to evaluate the predictive performance of the model.Methods:Patients with non-acute SICAO received endovascular therapy at 8 comprehensive stroke centers in China were included retrospectively. They were randomly assigned to a training set and a validation set. In the training set, the least absolute shrinkage and selection operator (LASSO) algorithm was used to screen important variables, and a decision tree prediction model was constructed based on CART algorithm. The model was evaluated using the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow goodness-of-fit test and confusion matrix in the validation set.Results:A total of 511 patients with non-acute SICAO were included. They were randomly divided into a training set ( n=357) and a validation set ( n=154) in a 7:3 ratio. The successful recanalization rates after endovascular therapy were 58.8% and 58.4%, respectively. There was no statistically significant difference ( χ2=0.007, P=0.936). A CART decision tree model consisting of 5 variables, 5 layers and 9 classification rules was constructed using the six non-zero-coefficient variables selected by LASSO regression. The predictive factors for successful recanalization included fewer occluded segments, proximal tapered stump, ASITN/SIR collateral grading of 1-2, ischemic stroke, and a recent event to endovascular therapy time of 1-30 d. ROC analysis showed that the area under curve of the decision tree model in the training set was 0.810 (95% confidence interval 0.764-0.857), and the optimal cut-off value for predicting successful recanalization was 0.71. The area under curve in the validation set was 0.763 (95% confidence interval 0.687-0.839). The accuracy was 70.1%, precision was 81.4%, sensitivity was 63.3%, and specificity was 79.7%. The Hosmer-Lemeshow test in both groups showed P>0.05. Conclusion:Based on the type of ischemic event, the time from the latest event to endovascular therapy, proximal stump morphology, the number of occluded segments, and the ASITN/SIR collateral grading constructed the decision tree model can effectively predict successful recanalization after non-acute SICAO endovascular therapy.

Objective:To investigate the relationship between caspase activation and recruitment domain 8 ( CARD8) gene rs2043211 polymorphism and the age of onset for large artery atherosclerosis (LAA) stroke in Chinese Han population. Methods:Based on Nanjing Stroke Registry Program, patients with LAA stroke from January 2010 to December 2014 were included retrospectively and genotyped. Grouping according to different genetic models, the age distribution among different genotypes were compared, and Kaplan-Meier curve was used to compare the age-related cumulative non-onset frequency of the patients with different genotypes.Results:A total of 738 patients were admitted and 717 (97.15%) were successfully genotyped and included in the study. There were no significant differences in age, sex, body mass index, hypertension, diabetes, hyperlipidemia, smoking and drinking history among the genotype groups. Patients with T allele had earlier onset under codominant model (AA genotype: 61.68 years, 95% confidence interval [ CI] 59.92-63.45 years; AT genotype: 60.51 years, 95% CI 59.41-61.60 years; TT genotype: 60.44 years, 95% CI 58.96-61.92 years). Kaplan-Meier curve analysis showed that there was a significant difference in the age-related cumulative non-onset frequency between patients with different genotypes (log-rank test, P=0.041). Similar results in the female was observed in the stratified analysis (log-rank test, P=0.001). Conclusions:T allele of CARD8 rs2043211 is associated with the early age of onset of large artery atherosclerosis stroke in Chinese Han population, especially in female patients.

Objective To evaluate the effects of posterior decompression,subtotal vertebrectomy,intervertebral fusion and internal fixation through unilateral pedicle in the treatment of severe throracolumar fractures.Methods A retrospective study was conducted to evaluate the outcomes of 11 patients with severe thmracolumar fracture who had been treated with posterior decompression,subtotal vertebrectomy,intervertebral fusion and internal fixation through unilateral pedicle at our department from March 2009 to March 2012.They were 10 males and one female,aged from 20 to 42 years (mean,31.1 years).According to AO classification,there were 2 cases of type B1,3 cases of type B2 and 6 cases of type C2.The Gaines scores for anterior column stability were more than 6 in all.All the patients had neurological disorders.The fracture reduction,fusion of bone grafting and recovery of neurological function were followed up.Results Their operation time ranged from 200 to 300 minutes,averaging 243.2 minutes;the amount of bleeding ranged from 800 to 1,600 mL,averaging 1,023.3 mL.No deterioration of neurological symptoms or no new neurological symptoms were observed postoperatively.Cerebrospinal fluid leakage was found in 3 patients who healed spontaneously following extubation,compressive dressing and elevation of the bed end.All the patients were followed up for 12 to 30 months (average,18.3 months).A loosened screw cap was found in one patient after 6 months who had no discomfort and received no treatment.X-ray films showed satisfactory fracture reduction and fine bone grafting locations.No spinal canal stenosis was found by CT scans.It took 3 to 6 months (mean,4.3 months) for the bone grafts to get fused.No height loss of the injured vertebrae was no found at the last follow-ups.No functional recovery was found in the patients with complete spinal cord injury whose Frankle grade remained A.The spinal function recovered from Frankel grade B to grade D in one patient with incomplete spinal injury.Of the 8 patients with medullary cone injury,7 recovered Frankle grade E and one Frankle grade D.The symptoms were relieved in one patient with simple cauda equine injury.Conclusion Posterior decompression,subtotal vertebrectomy,intervertebral fusion and internal fixation through unilateral pedicle is an effective alternative for treatment of severe throracolumar fractures,because it can lead to satisfactory fracture reduction,thorough decompression of the spinal canal and good reconstruction of spinal stability.

BACKGROUND:At present, liver transplantation is the only way to cure end-stage liver disease, but the complications after transplantation is stil an important factor of affecting the long-term survival of patients who received orthotopic liver transplantation, therefore it is necessary to establish a stable animal transplantation model. OBJECTIVE:To establish rat models of orthotopic liver transplantation. METHODS:After inhalation anesthesia with ether, 204 SD rats were perfused with 2-4℃ Ringer’s solution through the abdominal aorta. In order to reduce warm ischemia of the liver, the liver was not turned over before perfusion. The suprahepatic inferior vena cava was cut off along the phrenic ring after perfusion. No further trimming was needed when dressing, so as not to damage the vena cava. The donor liver was removed and preserved in 4℃Ringer’s liquid. The receptor liver was cut off and alogeneic orthotopic liver transplantation was performed using modified two-cuff method. After transplantation, rats could automaticaly turn over and drink water. Surviving more than 3 days is regarded as a successful transplantation. RESULTS AND CONCLUSION:102 liver transplantations were performed in 204 rats, with 86 rats surviving more than 3 days. The success rate of transplantation was 84%. The results demonstrate that rat models of orthotropic liver transplantation can be constructed successfuly through improving techniques.

Objective There is little research on the relationship of gene polymorphism of CYP2C19 and clopidogrel response after percutaneous transluminal angioplasty and stenting ( PTAS) in patients with ischemic cerebrovascular disease.The study aimed to investigate the relationship between gene polymorphism and high on-treatment platelet reactivity ( HTPR ) after PTAS and 6 months of regular dual antiplatelet administration in patients. Methods A total of 145 Chinese patients treated with PTAS in our de-partment from January 2011 to March 2014 were enrolled in this study.According to the gene sequencing, patients were divided into wild-type group(CYP2C19*1/*1,69 cases) and mutation group(heterozygous mutation CYP2C19*1/*2 and homozygous mutation CYP2C19*2/*2, 76 cases).Patients received a 100mg/d aspirin and 75mg/d clopidogrel maintenance dose (MD) as dual anti-platelet therapy after PTAS.The clopidogrel inhibition effect was measured by thrombelastography ( TEG) system 6 months after PTAS. Routine cerebral artery digital subtraction angiography was applied to evaluate whether there was restenosis in stent and logistic regres-sion analysis was used to analyze the influential factors of HTPR after PTAS and clopidogrel adminstration. Results After 6 months'regular administration of clopidogrel after PTAS, the platelet adenosine diphosphate ( ADP ) receptor inhibition rates in wild-type group, heterozygous mutation and homozygous mutation group were respectively (58.43 ±21.98)%, (47.80 ±22.93)%, (37.53 ± 21.84)%.The platelet ADP receptor inhibition rate was significantly decreased compared with wild-type group(P=0.001).Carriers of at least one CYP2C19 loss-of-function ( LOF) allele had a higher frequency of clopidogrel HTPR (35.5% vs17.4 % for patients with and without LOF alleles, respectively;P=0.014) .Using multivariate logistic regression analysis, the carriage of CYP2C19 LOF alleles was an independent predictor of the post-procedure HTPR (OR=2.356, 95% CI:1.053-5.272, P=0.037).The rate of ISR was remarkably higher in patients with at least one CYP2C19*2 alleles compared with wild-type patients(11.8%vs 1.4%, P=0.019) . Conclusion In patients with ischemic cerebrovascular disease, the CYP2C19 LOF allele had significant impact on post-procedure clopidogrel HTPR and the prognosis of ISR after PTAS.

Objective To sum up the clinical characteristics of children with severe pneumonia,and in order to improve the dingnosis treatment and prognosis.Methods Restrospective analysis was carried out on the clinical manifestations signs basic diseases etiology check imaging of children with severe pneumonia who had been in the hos-pital for ten years,and with the same period,120 children with common pneumonia in hospital were compared with and analyzed.Results Among 193 case,boys was 127 cases,girls was 63 cases,with males to females rate of 21.115 cases(60.0%)were aged 0 year to 3 years old.The onset of 125 cases(65.0%)were winter and spring.All patients had fever 98 cases(50.8%)with high temperature of 39 -41℃,all patients suffered from respiratory symp-toms,including 183 cases(94.8%)with cough,113 cases(58.5%)with breathing.All patients suffered from diffi-cult breathing,shortness of breath on cyanosis(=cyanopathy),105 cases(54.3%)with wet lung rate,152 cases (78.7%)with respiratory insufficiency on respiratory failure,12 cases(6.2%)with acute respiratory distress syn-drome,163 cases (84.5%)with heart failure,67 cases (34.7%)with abdominal distension,23 cases needed mechanical ventication.91 cases(47.2%)with antibodies positime for mycoplasma pneumoniae infection when testing servm virns antibody and respiratory virus antigen.20 cases(10.4%)were found to have positive antibody,conduc-ting fluid culture,cultivate a positive strains of 31 cases(16.1%),including 7 cases(3.6%)of psendomonas aerngi-nosa,6 cases(3.1%)of eschericria coli,6 cases(3.1%)of klebsiella pneumoniae,5 cases(2.6%)of enterobacter cloacae,3 cases(1.6%)of streptococlus pneumoniae,2 cases(1.0%)of viridans streptococci,1 case(0.5%)of hemolytic staphylococci,1 case(0.5%)of radiation agrobacterium,2 cases(1.0%)of candida mycoderma bacteria which was fungi.The imaging indicated.106 cases(54.9%)presented as lobi pulmonis or segmental large patches of dense increased shadom or pulmonary parenchymal inflammatory lesions the performance of lung interstitial inflammatory lesionsl(such as increased lung markings,fuzzy and with flocculant shadow etc)were 87 cases(33.2%).30 cases (15.5%)suffered from pleural effusion,18 cases(9.3%)suffered a telectasis with in the chest,16 cases(8.3%) suffered from empyema.Conclusion Children with severe pneumonia had prone to heart failure respiratory failure, complication.The clinical manifestations of severe pneumonia is severe.Clinically suspected severe.Pneumonia should complete etiological and chest radiographic examination for early diagnosis and treatment.

Objective Neuroinflammation following traumatic brain injury (TBI) may give rise to neurodisorder.This study aimed to investigate the effect of intranasal delivery of nerve growth factor ( NGF) on neuroinflammation following TBI and its action mechanism in rats. Methods Thirty-six male adult Sprague-Dawley rats were equally divided into a sham , a TBI, and a TBI+NGF group.The rats in the TBI +NGF group were treated with NGF intranasally at 12 and 24 hours after TBI.The levels of IL-1βand TNF-αin the injured cerebral cortex were detected by ELISA , the DNA-binding activity of NF-κB evaluated by EMSA , and the expres-sion of amyloid-β( Aβ42 ) determined by Western blot . Results NGF attenuated the inflammation following TBI .Compared with the TBI group, the level of IL-1βwas obviously decreased in the TBI +NGF group at 12 hours (70.65 ±3.10 vs 37.51 ±1.92) and 24 hours (68.85 ±8.10 vs 36.23 ±2.99, P<0.05), and so was that of TNF-α(47.12 ±7.38 vs 27.63 ±5.77 and 56.15 ±11.20 vs 29.94 ±8.62, P<0.05).The DNA-binding activity of NF-κB was reduced to 111.62 ±0.49 and 131.52 ±0.88, and the expression of Aβ42 to 0.23 ±0.008 and 0.52 ±0.004 at 12 and 24 hours respectively after treatment with NGF , both with statistically significant differences from the TBI group (P<0.05). Conclusion Intranasal administration of NGF attenuates TBI-induced neuroinflamma-tion in rats, which may be associated with its regulatory effect on the Aβ42/NF-κB signaling pathway .

Objective To investigate the effects of the distribution characteristics of cerebral artery stenosis and the associated risk factors in patients with ischemic cerebrovascular disease.Methods The demographic data and vascular risk factors in patients with ischemic cerebrovascular disease who performed aortic arch and cerebral angiography were analyzed retrospectively.The patients were divided into intracranial lesion,extracranial lesion and extra-and intracranial lesion groups according to the lesion sites.The demographic data and vascular risk factors in all groups were compared.Results A total of 1272 patients were enrolled,and 1028 (80.8％) had cerebral artery stenosis or occlusion,in which 342 (33.3％) were intracranial lesions,330(32.1％) were extracranial lesions,and 356 (34.6％) were extra-and intracranial lesions.The mean age of the intracranial lesion group was significantly lower than that of the other 2 groups (F =41.995,P =0.000).There were significant differences in the constituent ratios of sex (x2 =10.602,P =0.005),hypertension (x2 =11.316,P =0.003),and diabetes (x2 =13.465,P =0.001) among all groups.There were significant differences in the distribution of extra-and intracranial artery stenosis among different age groups (P =0.001).Intracranial lesions were mainly in the youth and middle-aged groups,and extra-and intracranial lesions in the old age group were more common.Multivariate logistic regression analysis showed that age,hypertension,diabetes and history of transient ischemic attack were associated with the simple intracranial lesions and extra-and intracranial lesions,and the simple extracranial lesions were only associated with advanced age and hypertension.The 1028 patients with vascular stenosis or occlusion affected 2732 vessels,including 1759 vessels (64.4％) in anterior circulation and 973 (35.6％) in posterior circulation.The mild,moderate and severe stenosis in anterior circulation were most common in extracranial internal carotid artery,and the occlusion was most common in middle cerebral artery.The various degrees of lesions in posterior circulation were most common in extracranial internal carotid artery.Conclusions There were significant differences in patients with cerebrovascular lesions at different sites in sex,age,as well as in the incidences of hypertension and diabetes.Age,hypertension,diabetes and the history of transient ischemic attack were the independent predictive factors for the distribution of cerebral atherosclerotic lesions.