AIM: To evaluate the expression of tumor necrosis factor-α(TNF-α)in the iris and ciliary body of Wistar rats in the endotoxin-induced uveitis(EIU), and the effect of Xuebijing injection on its expression.METHODS:A total of 65 Wistar rats were randomly divided into three groups: Group A(normal saline, n=5), Group B(normal saline+endotoxin-injected, n=30), and Group C(Xuebijing+endotoxin-injected, n=30). The EIU model was induced in Wistar rats of the groups B and C by injecting LPS into the plantar surfaces of the hind feet, and normal saline(15 mL/kg)or Xuebijing(15 mL/kg)were intraperitoneally administered 30 min before LPS administration. The rats of the groups B and C were further divided into 6 subgroups after LPS injection, including 6, 12, 18, 24, 48, and 72 h subgroups, with 5 rats in each group. Furthermore, the intraocular inflammation of the rats was observed at each time above, the number of infiltrating cells in the aqueous humor was counted, and the pathological changes were observed in the iris and ciliary body of rats using hematoxylin and eosin(HE)staining. TNF-α expression in iris and ciliary tissue at different postoperative time points was evaluated using immunohistochemistry.RESULTS: Clinical observations indicated no signs of uveitis in the group A, signs of uveitis were observed in the group B. Both iris symptoms and damage were significantly reduced in the group C compared to the group B(P<0.01). Cell counts in the aqueous humor revealed no inflammatory cells in the group A, while the number of aqueous humor cells in the group C was significantly reduced compared to Group B(P<0.01). HE staining revealed no cellular infiltration in the group A. In the group B, some cellular infiltration was observed in the eyes at 6 h post-LPS exposure. The number of infiltrating cells increased over time, peaked at 24 h, and gradually declined thereafter. In the group C, cell infiltration was not obvious at 6 h, few at 24 h, and nearly disappeared by 48 h. Immunohistochemical staining showed higher TNF-α expression in the ciliary body and iris in the group B than in the group A(P<0.01). Compared to the group C, TNF-α expression in the group B was significantly upregulated following LPS injection(P<0.01).CONCLUSION:TNF-α expression was elevated in EIU rats, and there was a positive correlation between its mean optical density ratio and inflammation degree. Moreover, Xuebijing injection could alleviate inflammation response through the reduction of TNF-α levels.

BACKGROUND:There are many kinds of cell media with different components,which have a great influence on cell growth.Several studies in and outside China have used serum-free media containing fetal bovine serum for in vitro amplification culture,but the use of media containing human peripheral blood serum to directionally induce human umbilical cord mesenchymal stem cells to neural stem cells and human peripheral blood serum to promote neural stem cells to differentiate into other nerve cells,so far there are few relevant studies. OBJECTIVE:To observe the feasibility of inducing human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells. METHODS:(1)Human umbilical cord mesenchymal stem cells were cultured in DMEM/F-12 culture medium containing 10%human peripheral blood serum by volume fraction.Flow cytometry analysis was performed at the third passage to identify surface markers and alizarin red staining was used to detect osteogenic differentiation function.(2)The third-generation human umbilical cord mesenchymal stem cells were induced into neural stem cells using DMEM/F-12 medium containing 0.5%N2,1.5%B27,20 ng/mL basic fibroblast growth factor,and 20 ng/mL epidermal growth factor,and their surface markers were identified.(3)Well-growing human umbilical cord mesenchymal stem cell-derived neural stem cells were taken to prepare a single cell suspension.They were evenly inoculated into 96-well plates and incubated with DMEM/F-12 culture medium containing 10%human peripheral blood serum for 8 days.Then,hematoxylin-eosin staining,microtubule-associated protein 2,and glial fibrillary acidic protein immunofluorescence staining were performed to detect the differentiation of neural stem cells into other neural cells. RESULTS AND CONCLUSION:(1)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum grew in a spiral-like pattern and were distributed in multiple layers,with directional arrangement.The surface of human umbilical cord mesenchymal stem cells highly expressed CD44,CD105,CD29,and CD73.Cells stained with alizarin red showed a color reaction.(2)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum could be induced to differentiate into neural stem cells,and the surface of neural stem cells was highly expressed with CD44,CD105,CD29,CD73,Nestin,NF-L,and GALC.(3)On day 8 of induced differentiation of neural stem cells,after staining with hematoxylin and eosin,it was found that the protruding protrusions were longer,with more branches and adjacent cells connected,presenting typical neural cell morphology.Immunofluorescence staining for microtubule-associated protein 2 and glial fibrillary acidic protein was positive.It is concluded that human umbilical cord mesenchymal stem cells cultured by human peripheral blood serum can be directly induced to differentiate into neural stem cells.Under the action of human peripheral blood serum,neural stem cells can differentiate into other neural cells as the culture time prolongs.

Objective:pH low insertion peptide (pHLIP)-variant 7 (var7)-fluorescein isothiocyanate (FITC) was used to explore an accurate imaging tool that targeted burn wounds to better perform burn debridement.Methods:Twelve rat models of burn wound were established and pHLIP-var7-FITC with different concentrations (0.5, 1.5 and 2.0 mg/ml) were injected from the rat tail vein for in vivo fluorescence imaging. By determining the concentration of fluorescent conjugates to the burn wound, the scope of wound injury necrosis was judged by combining pathological sections, and its residue and toxicity in important organs such as heart, liver, kidneys, and brain were detected. The Kruskal-Wallis rank sum test, Bonferroni correction method and one-way analysis of variance were used for data analysis. Results:Within 24 h, the fluorescence photons per unit area of the burn wound in the group of 0.5 mg/ml, 1.5 mg/ml and 2.0 mg/ml were 1.49(1.31, 1.65), 2.46(1.88, 2.68), 2.77 (1.94, 3.10)×10 7 p·s -1·cm -2·Sr -1, with significant differences in the overall distribution of fluorescence photons ( H=73.55, P<0.001). The fluorescence intensity was stronger in the group with higher concentration, but with no significant difference in the number of fluorescence photons between the group of 1.5 mg/ml and 2.0 mg/ml ( P=0.263, Bonferroni correction method). At 14 time points (0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, 12, 24 h), there was no significant difference in the overall mean of fluorescence photons ( F=1.04, P=0.419), and the tissue with burn necrosis seen in tissue sections was highly consistent with the fluorescence imaging region. There was no obvious fluorescence residue in the heart, liver, kidney and brain sections. Conclusion:In superficial second-degree burn tissue, pHLIP-var7-FITC can accurately target and gather on the burn wound within 24 h, showing a clear boundary between burn tissue and normal tissue, which can assist clinical surgical debridement to determine the extent of injury.

Objective:To investigate the clinical outcome of endovascular treatment vs. drug treatment in patients with symptomatic non-acute long-segment occlusion of the internal carotid artery. Methods:Based on prospective cohort registration research data, patients with symptomatic non-acute long-segment occlusion of internal carotid artery were retrospectively included. They were divided into a drug treatment group and an endovascular treatment group according to the actual treatment received. The latter was further divided into a successful recanalization group and an unsuccessful recanalization group. The endpoint events included ipsilateral ischemic stroke, any stroke, and all-cause death. Multivariate logistic regression analysis was used to compare the endpoint events between groups during the perioprocedural period (within 30 days), and multivariate Cox proportional hazards model was use to compare the endpoint events between the groups during the long-term follow-up. Results:A total of 684 patients were included, of which 570 (83.33%) were male, median aged 63 years (interquartile range, 56-70 years). Three hundred and fifty-three patients (51.6%) received drug treatment; 331 (48.4%) received endovascular treatment, of which 161 (48.6%) had successful recanalization. The median follow-up time was 1 223 days (interquartile range, 646.5-2 082 days), with 109 patients (15.9%) experiencing stroke recurrence events (including 87 ipsilateral ischemic stroke) and 78 (11.4%) experiencing all-cause mortality. The risk of any stroke during the perioprocedural period in the successful recanalization group was significantly higher than that in the drug treatment group (odds ratio 3.679, 95% confidence interval 1.038-13.036; P=0.044), but the risk of ipsilateral ischemic stroke recurrence (risk ratio 0.347, 95% confidence interval 0.152-0.791; P=0.012) and all-cause mortality (risk ratio 0.239, 95% confidence interval 0.093-0.618; P=0.003) during the long-term follow-up were significantly lower than those in the drug treatment group. Conclusions:In patients with symptomatic non-acute long-segment occlusion of the internal carotid artery, endovascular treatment can increase the risk of stroke recurrence within 30 days, but successful recanalization can reduce the risks of long-term ipsilateral ischemic stroke recurrence and all-cause mortality.

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.

Brain functional network changes over time along with the process of brain development, disease, and aging. However, most of the available measurements for evaluation of the difference (or similarity) between the individual brain functional networks are for charactering static networks, which do not work with the dynamic characteristics of the brain networks that typically involve a long-span and large-scale evolution over the time. The current study proposes an index for measuring the similarity of dynamic brain networks, named as dynamic network similarity (DNS). It measures the similarity by combining the "evolutional" and "structural" properties of the dynamic network. Four sets of simulated dynamic networks with different evolutional and structural properties (varying amplitude of changes, trend of changes, distribution of connectivity strength, range of connectivity strength) were generated to validate the performance of DNS. In addition, real world imaging datasets, acquired from 13 stroke patients who were treated by transcranial direct current stimulation (tDCS), were used to further validate the proposed method and compared with the traditional similarity measurements that were developed for static network similarity. The results showed that DNS was significantly correlated with the varying amplitude of changes, trend of changes, distribution of connectivity strength and range of connectivity strength of the dynamic networks. DNS was able to appropriately measure the significant similarity of the dynamics of network changes over the time for the patients before and after the tDCS treatments. However, the traditional methods failed, which showed significantly differences between the data before and after the tDCS treatments. The experiment results demonstrate that DNS may robustly measure the similarity of evolutional and structural properties of dynamic networks. The new method appears to be superior to the traditional methods in that the new one is capable of assessing the temporal similarity of dynamic functional imaging data.
Aging/physiology* ; Brain/physiology* ; Brain Mapping ; Humans ; Magnetic Resonance Imaging/methods* ; Nerve Net/physiology* ; Transcranial Direct Current Stimulation/methods*

Objective:To identify the morphological features and clinical significance of intra-vascular thrombus in carotid stenosis using optical coherence tomography (OCT).Methods:Twelve patients with carotid artery thrombosis detected by OCT assessment in Jinling Hospital between January 2017 and January 2020 were included. Serial area measurements within the athero-thrombotic target lesion were performed to evaluate the OCT-thrombus score, length, area and volume. The clinical data and plaque morphological features were also assessed.Results:Among the 12 patients demonstrating thrombus on OCT, eight patients presented with white thrombus, two patients presented with red thrombus, and another two patients displayed both white thrombus and red thrombus. OCT-thrombus scores were 1-32. The OCT-thrombus score was correlated to the OCT-thrombus volume ( ρ=0.739, P=0.006) and the thrombus length ( ρ=0.932, P<0.001). All lesions were presented with fibrous cap disruption, and 10 lesions were presented with thin-cap fibroatheroma. In view of the OCT findings, all patients received carotid balloon angioplasty and stent implantation. During an average follow-up of 14.2 months, none of the 12 patients had fatal stroke or recurrent ischemic stroke. Conclusions:OCT can be used to assess intra-carotid thrombus and its more detailed morphological characteristics, offering more possibilities in quantitative analysis of thrombus burden.

Objective:To have a better understanding of congenital leukemia by summarizing its clinical features and prognosis.Methods:This study retrospectively recruited 10 neonates with congenital leukemia treated in Beijing Children's Hospital, Capital Medical University from January 2006 to December 2018. Clinical data including clinical manifestations, laboratory examinations, treatment and follow-up were described.Results:The 10 patients consisted of six boys and four girls. In all cases, symptoms presented within 11 d after birth. The admission complaints were jaundice ( n=4), polypnea ( n=3), fever ( n=2) and rash ( n=2). Physical examinations of the 10 patients showed eight with splenomegaly, seven with hepatomegaly and seven with petechia/skin rash. All patients had significantly increased white blood cell count (from 45.8×10 9/L to 553.0×10 9/L), complicated by different degrees of anemia and thrombocytopenia. By bone marrow biopsy, two cases were diagnosed as acute lymphoblastic leukemia and the other eight cases as acute myeloid leukemia. All cases refused chemotherapy on the preliminary diagnosis. Three cases lost follow-up and six died within two months after discharge requested by their parents. One baby had spontaneous remission, but relapsed two years later. Complete remission was achieved after strict management and no relapse was reported until ten years old. Conclusions:Congenital leukemia is a severe condition with high mortality. Some cases may achieve spontaneous remission, but long-term follow-up is needed.

Objective:Acute type A dissection with mitral regurgitation was very rare. To sum up our experience of surgical treatment of acute type A dissection with mitral regurgitation.Methods:From October 2010 to January 2017, 17 patients with acute type A dissection with mitral regurgitation underwent surgical treatment. There were 12 males, 5 females; mean age, (38±11) years(range, 18-59 years). Hypertension was noted in 9 patients, Marfan syndrome in 2 patients, renal dysfuction in 3 patients, cardiac dysfunction in 3 patients, lower limb ischemia was observed in 2 patients and dilated cardiomyopathy in one patient. Preoperative abdominal aortic replacement was observed in one case.Results:Concomitant procedures included Bentall procedure in 16 patients, aortic arch operation in 15 patients, tricuspid valve plasty in 3 patients, coronary artery bypass grafting in 2 patients and asceding aorta - femoral artery bypass in one patient. 11 patients required mechanical ventilation for <24 hours, 3 cases for <48 hours and 3 subjects for > 48 hours. Continuous renal replacement therapy was required in 3 patients, re-operation in one patient and partial pericardial excision in one patient. One patient was out of follow-up. The remaining had a normal life during follow-up.Conclusion:Repair of acute type A dissection with mitral regurgitation carried with a relatively high mortality and morbidity. Under better protection of heart and cerebrum, it obtained accepted surgical results in patients with this lesion

Objective To elucidate the mechanism underlying eletroacupuncture's (EA's) amply-documented analgesic effect.To observe its effect on the pain threshold and on the expression of P2X4 receptor and microglia activation in the spinal cords of rats with neuropathic pain.To demonstrate whether or not interfering with A1 receptors and P2X4 receptors at the same time could enhance the analgesic effect.Methods A total of 40 SpragueDawley rats weighing 150 to 180 g were randomly divided into a sham group,a CCI group,an EA group,a 2-chloroN(6)-cyclopentyladenosine (CCPA) group and a CCPA+EA group,each of 8.Chronic constriction injury (CCI)was induced successfully in the rats of all groups except the sham group.Five days later,EA and 20 μL injections of 0.1 mm/L CCPA were applied to the rat analogues of the Zusanli (ST36) and Yanglingquan (GB34) acupoints once a day for 15 days for the rats in the appropriate groups.The mechanical withdrawal threshold (MWT) and the thermal withdrawal latency (TWL) were measured before the CCI operation and 20 days afterward.L4-L6 spinal cord tissue was then resected and the fluorescence intensity of P2X4 and OX42 receptors was detected using double label immunohistochemical staining.The correlation between the mean fluorescence intensity and the pain threshold gap was analyzed.Results The average MWT and TWL of the CCI group were significantly lower than in the other four groups.The expression of P2X4 receptor and OX42 in the spinal cord increased significantly in the CCI group compared to the other four groups.There was significant correlation between the mean fluorescence intensity of P2X4 and OX42 receptors and the gap in pain threshold,with correlation coefficients of 0.907 and 0.717 respectively.Conclusion P2X4 receptor and microglia activation might be involved in the development of neuropathic pain.CCPA and EA can inhibit the activation of microglia and reduce the activity of P2X4 receptors.The interaction between A1 receptors and P2X4 receptors can strengthen the analgesic effect of EA.