Interactions between brain-resident and peripheral infiltrated immune cells are thought to contribute to neuroplasticity after cerebral ischemia. However, conventional bulk sequencing makes it challenging to depict this complex immune network. Using single-cell RNA sequencing, we mapped compositional and transcriptional features of peri-infarct immune cells. Microglia were the predominant cell type in the peri-infarct region, displaying a more diverse activation pattern than the typical pro- and anti-inflammatory state, with axon tract-associated microglia (ATMs) being associated with neuronal regeneration. Trajectory inference suggested that infiltrated monocyte-derived macrophages (MDMs) exhibited a gradual fate trajectory transition to activated MDMs. Inter-cellular crosstalk between MDMs and microglia orchestrated anti-inflammatory and repair-promoting microglia phenotypes and promoted post-stroke neurogenesis, with SOX2 and related Akt/CREB signaling as the underlying mechanisms. This description of the brain's immune landscape and its relationship with neurogenesis provides new insight into promoting neural repair by regulating neuroinflammatory responses.
Humans ; Ischemic Stroke ; Brain/metabolism* ; Macrophages ; Brain Ischemia/metabolism* ; Microglia/metabolism* ; Gene Expression Profiling ; Anti-Inflammatory Agents ; Neuronal Plasticity/physiology* ; Infarction/metabolism*

Objective:To observe the short-and mid-term efficacy of left subclavian artery(LSA) laser in situ fenestration combined with arch debranching surgery for aortic arch reconstruction in patients with Stanford type A aortic dissection aged 60 years and above. Methods:This is a retrospective cohort study. A total of 41 Stanford type A aortic dissection patients aged 60 years and above who received combined surgery in Department of Endovascular Surgery, the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2020 were retrospectively analyzed. There were 25 males and 16 females, aged (67.3±5.9)years(range: 60 to 75 years). Among them, 19 patients underwent LSA laser in situ fenestration combined with arch debranching surgery(combined surgery group) and 22 patients underwent hybrid aortic arch debranching surgery(non-combined surgery group). Independent sample t test, χ2 test and Fisher exact probability method were used to compare the clinical characteristics of the two groups. Kaplan-Meier method was used for survival analysis, and the 5-year survival rate of the two groups was compared by Log-rank test. Results:Body mass index in the combined operation group was significantly higher than that in the non-combined operation group ((27.1±1.6)kg/m 2vs.(26.9±1.9)kg/m 2; t=2.766, P=0.006), and the difference was statistically significant. There was no statistical significance in the comparison of other general data (all P>0.05). The operation time ((321.3±11.4) minutes vs. (329.6±7.3)minutes; t=-2.733, P=0.010) and LSA reconstruction time ((32.4±3.0)minutes vs. (42.4±6.0)minutes; t=-6.842, P<0.01) in the combined operation group were significantly shortened, and the difference was statistically significant. The rate of LSA reconstruction in the combined operation group (100% vs. 72.7%; P=0.023) was significantly higher than that in the non-combined operation group, and the difference was statistically significant. There were no significant differences in the incidence of pulmonary infection, unplanned second operation, continuous renal replacement therapy, neurological complications and the in-hospital mortality between the two groups. Compared with the non-combined surgery group, the total complication rate related to LSA reconstruction was significantly lower in the combined surgery group (0 vs. 27.3%; P=0.023). Kaplan-Meier survival analysis showed that there was no difference in 5-year survival rate between the combined operation group and the non-combined operation group (84.2% vs. 77.3%; χ2=0.310, P=0.578). Conclusion:Laser in situ fenestration of the LSA combined with arch debranching surgery to reconstruct the aortic arch can significantly shorten the operation and LSA reconstruction time in patients aged 60 years and above with Stanford type A aortic dissection, improve the success rate of LSA reconstruction, and reduce the occurrence rate of LSA reconstruction complications.

Objective:To observe the short-and mid-term efficacy of left subclavian artery(LSA) laser in situ fenestration combined with arch debranching surgery for aortic arch reconstruction in patients with Stanford type A aortic dissection aged 60 years and above. Methods:This is a retrospective cohort study. A total of 41 Stanford type A aortic dissection patients aged 60 years and above who received combined surgery in Department of Endovascular Surgery, the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2020 were retrospectively analyzed. There were 25 males and 16 females, aged (67.3±5.9)years(range: 60 to 75 years). Among them, 19 patients underwent LSA laser in situ fenestration combined with arch debranching surgery(combined surgery group) and 22 patients underwent hybrid aortic arch debranching surgery(non-combined surgery group). Independent sample t test, χ2 test and Fisher exact probability method were used to compare the clinical characteristics of the two groups. Kaplan-Meier method was used for survival analysis, and the 5-year survival rate of the two groups was compared by Log-rank test. Results:Body mass index in the combined operation group was significantly higher than that in the non-combined operation group ((27.1±1.6)kg/m 2vs.(26.9±1.9)kg/m 2; t=2.766, P=0.006), and the difference was statistically significant. There was no statistical significance in the comparison of other general data (all P>0.05). The operation time ((321.3±11.4) minutes vs. (329.6±7.3)minutes; t=-2.733, P=0.010) and LSA reconstruction time ((32.4±3.0)minutes vs. (42.4±6.0)minutes; t=-6.842, P<0.01) in the combined operation group were significantly shortened, and the difference was statistically significant. The rate of LSA reconstruction in the combined operation group (100% vs. 72.7%; P=0.023) was significantly higher than that in the non-combined operation group, and the difference was statistically significant. There were no significant differences in the incidence of pulmonary infection, unplanned second operation, continuous renal replacement therapy, neurological complications and the in-hospital mortality between the two groups. Compared with the non-combined surgery group, the total complication rate related to LSA reconstruction was significantly lower in the combined surgery group (0 vs. 27.3%; P=0.023). Kaplan-Meier survival analysis showed that there was no difference in 5-year survival rate between the combined operation group and the non-combined operation group (84.2% vs. 77.3%; χ2=0.310, P=0.578). Conclusion:Laser in situ fenestration of the LSA combined with arch debranching surgery to reconstruct the aortic arch can significantly shorten the operation and LSA reconstruction time in patients aged 60 years and above with Stanford type A aortic dissection, improve the success rate of LSA reconstruction, and reduce the occurrence rate of LSA reconstruction complications.

Objective:To prepare the whole bladder acellular matrix (BAM) using the self-designed perfusion decellularization system, and evaluate the feasibility of constructing the tissue engineering bladder with the adipose-derived stem cells (ADSCs).Methods:This study was conducted from October 2020 to April 2021. The self-designed perfusion decellularization system was used, and four different decellularization protocols (group A, group B, group C and group D) were formulated, according to the flow direction of the perfusate and the action time of different decellularization solutions. Among them, the urethral orifice of the bladder tissue was used as the outflow tract of the perfusion fluid in groups A and B. The top of the bladder was cut off and used as the outflow tract of the perfusion fluid in groups C and D. In groups A and C, 1% Triton X-100 was treated for 6 h, and 1% sodium dodecyl sulfate (SDS) was treated for 2 h. In groups B and D, 1% Triton X-100 was treated for 7 h, and 1% sodium dodecyl sulfate (SDS) was treated for 1 h. In addition, the tissue in the normal bladder group was directly obtained from the natural bladder tissue, which did not require perfusion, cryopreservation and thawing. The fast and efficient decellularization protocol was screened out through HE, DAPI, Masson trichrome and Alcian Blue staining and quantitative analyses to prepare the whole bladder scaffold. The prepared BAM was used as the scaffold material, and the ADSCs were used as the seeding cells to construct the tissue engineering bladder. HE and DAPI staining were used to observe the distribution of ADSCs on the BAM.Results:HE and DAPI staining showed that there was no obvious nuclear residue in the group C. Masson trichrome and Alcian Blue staining showed that the collagen structure and glycosaminoglycan were well preserved in the group C. There was no significant difference in bladder wall thickness between the group C and the normal bladder group [(975.44±158.62)μm vs.(1 064.49±168.52)μm, P > 0.05]. The DNA content in the group C [(43.59 ±4.59) ng/mg] was lower than that in the normal bladder group, group A, group B and group D [(532.50±26.69), (135.17±6.99), (182.49±13.69) and(84.00±4.38)ng/mg], and the difference was statistically significant ( P<0.05). The collagen content [(10.98 ± 0.29)μg/mg] and glycosaminoglycan content [(2.30±0.18)μg/mg] in group C were not significantly different with those in the normal bladder group [(11.69±0.49) and (2.36±0.09)μg/mg, P>0.05]. Scanning electron microscopy showed that a large number of pore structures could be observed on the surface of the prepared BAM in groups A-D and were facilitated to cell adhesion. The isolated and cultured ADSCs were identified by flow cytometry to confirm the positive expression of CD90 and CD29, and the negative expression of CD45 and CD106. Live/dead staining and CCK-8 detection confirmed that the prepared BAM in the group C had no cytotoxicity. HE and DAPI staining showed that a large number of ADSCs were distributed on the surface and inside of the tissue engineering bladder. Conclusions:The whole bladder shape BAM prepared by the self-designed perfusion decellularization system could be used as the scaffold material for bladder tissue engineering, and the constructed tissue engineering bladder could be used for bladder repair and reconstruction.

Objective:To investigate the effect of tissue engineered bladder patch constructed by double-layer silk scaffold and adipose-derived stem cells (ADSCs) in the repair and reconstruction of bladder.Methods:This study was conducted from May 2020 to March 2021. The silk fibroin (SF) aqueous solution was obtained from silkworm cocoons, and a double-layer silk scaffold composed of silk fibroin film and silk fibroin sponge was further prepared. The rat ADSCs were isolated, cultured, and the ADSCs surface markers (CD29, CD90, CD45, CD106) were identified by flow cytometry. The ADSCs were planted on a double-layer silk scaffold to construct a tissue-engineered bladder patch. Thirty-six male SD rats were randomly divided into three groups: tissue engineered bladder patch group (SF-ADSCs group, n=15), double-layer silk scaffold group (SF group, n=15), control group ( n=6). The tissue engineered bladder patch (SF-ADSCs group) and double-layer silk scaffold (SF group) were wrapped on the omentum to promote vascularization. The vascularization was evaluated by HE and immunofluorescence staining. The wrapped tissue engineered bladder patch and double-layer silk scaffold were used to repair the defective bladder. In the control group (six rats), the incision was closed immediately after the bladder tissue fully exposed. At 4 weeks and 12 weeks after operation, the general morphology of bladder tissue and cystography were performed to evaluate the recovery of bladder morphology. After the graft was harvested, HE and Masson's trichrome staining and immunofluorescence staining were used to observe the regeneration of bladder wall tissue. Urodynamics was used to assess the recovery of bladder function at 12 weeks after operation. Results:The flow cytometry results confirmed that the isolated cells positively expressed CD29 and CD90, and there was no significant expression of CD45 and CD106. Gross observation and scanning electron microscope confirmed that the preparation of double-layer silk scaffold not only had a pore structure that was conducive to cell planting, but also had good toughness and was facilitated to surgical suture. The number (43.50±2.66) and area (0.73±0.03)% of vascular-like structures in the SF-ADSCs group after the omentum encapsulation was significantly higher than that in the SF group [(24.50±3.51), (0.55±0.05)%], and the difference was statistically significant ( P<0.05). At 4 weeks after bladder repair, the histological staining of the grafts in the SF-ADSCs and SF groups showed a large number of degraded fragments of double-layer silk scaffold. At 12 weeks, the morphology of the graft in the SF-ADSCs group showed uniform bladder morphology, which was similar to that of normal bladder tissue. Immunofluorescence staining showed that the continuous urothelial layer, abundant smooth muscle tissue, vascular structure and regenerated neurons could be observed in the SF-ADSCs group. Urodynamic test showed that the bladder maximum volume (0.74±0.03)ml and compliance (16.68±0.44)μl/cm H 2O in the SF-ADSCs group, which were better than that in the SF group [(0.47±0.05)ml, (14.89±0.37)μl/cm H 2O], but lower than that in the control group [(1.12±0.08)ml, (19.34±0.45)μl/cm H 2O], and the difference was statistically significant ( P<0.05). Conclusions:The tissue engineered bladder patch constructed with double-layer silk scaffolds and ADSCs could promote the morphological repair of bladder tissue, the regeneration of bladder wall structure and the recovery of bladder physiological function.

Objective:To evaluate the mid-term results of endovascular treatment for transplant renal artery stenosis (TRAS).Methods:The clinical and follow-up data of TRAS patients undergoing endovascular treatment at the First Affiliated Hospital of Zhengzhou University from Jan 2014 to Jan 2021 were retrospectively analyzed.Results:A total of 2 230 patients underwent kidney transplantation, 78 cases(3.6%) developed TRAS, among those 27 patients received endovascular treatment and followed-up from 12 to 80 months(mean 36 months). Thirteen patients (48.1%) underwent renal graft angiography and balloon dilatation, of which 2 patients underwent stent placement, 14 patients (51.9%) underwent renal graft angiography with balloon dilatation and stenting. The serum creatinine 2 weeks postoperatively and 12 months postoperatively were 127.6 μmol/L (47-220 μmol/L) and 103.4 μmol/L (63-166 μmol/L), respectively, significantly lower than the preoperative 217.1 μmol/L (98-541 μmol/L), ( P<0.05). Glomerular filtration rate (GFR) before surgery was 8.3-105.3 ml/min, 2 weeks and 12 months after surgery compared to 24.6-132.2 ml/min and 47.3-113.9 ml/min( P<0.05). The preoperative peak systolic velocity (PSV) of the transplanted renal artery during the systolic phase was 234 cm/s (75-457 cm/s), compared to 129 cm/s (52-290 cm/s) ( P<0.05) 2 weeks and 118 cm/s (57-300 cm/s) 12 months postoperatively ( P<0.05). During the follow-up period, 2 patients (7.4%) died of multiple organ failure. Conclusions:TRAS is the most common vascular complication after kidney transplantation. Endovascular treatment has a high success rate and low complication rate.

【Objective】 To explore the methods and safety of autologous peripheral hematopoietic stem cells collection in patients with sequential double transplantation of solid tumors and conduct efficacy analysis. 【Methods】 Peripheral blood stem cells were collected from 27 patients with solid tumors after routine mobilization of rhG-CSF and rhGM-CSF.A specific program was made for the patients.The condition and cooperation degree of children were comprehensively evaluated before cell collection, and a femoral venous catheterization was inserted to ensure the cells collected smoothly.A mononuclear cell collection(MNC) program was selected, and machine parameters were set based on the patient's low body weight.The number of mononuclear cell (MNC) and the CD34⁺ cell was detected by flow cytometry for retrospective analysis. 【Results】 A total of 73 cell collections were performed in 27 patients, and the number of mononuclear cells and CD34⁺ cells was 12.586(10.22~19.586)×10⁸/kg and 13.575(7.275~23.825)×10⁶/kg, respectively, which can meet the requirement of sequential double transplantation. No intoxication of citrate and other serious adverse reactions occurred, and the follow-up was generally in good condition. 【Conclusion】 The method is effective and safe for pediatric patients, even for pediatric patients with low weight. Sufficient stem cells can be collected for patients with solid tumors by this method to meet the requirement of sequential double transplantation.

Objective: To investigate the effect of casein kinase 2 interacting protein-1 (CKIP-1) on craniofacial soft tissues and hard tissues, to provide the basis for the study and treatment of craniomaxillofacial related diseases. Methods: 6-month- old male CKIP-1 knockout (KO) mice were selected as the experimental group, and wild-type (WT) mice were selected as the control group. The craniomaxillofacial hard tissues (parietal bone, nasal bone, incisors and molars) were analyzed through micro- CT, and the morphological changes of maxillofacial soft tissues (nasal cartilage, lip mucosa and tongue) were analyzed through HE staining and toluidine blue staining. Results: CKIP-1 negatively regulated bone mass of cancellous bone of cranial and maxillofacial bones and dentin mineralization. Compared with the WT mice, the thickness of the parietal baffle layer increased by 93% in KO mice, while cortical bone showed no significant difference between the two groups. The nasal cancellous bone thickness increased by 160% in KO-mice, while cortical bone showed no significant difference between the two groups; the enamel thickness was normal, but the pulp cavity became smaller and the dentin thickness increased by 48%. Compared with the WT mice, the HE staining and toluidine blue staining analyses of the soft tissues revealed that the thickness of the alar cartilage plate of KO mice increased by 57%, and local ossification was found within the cartilage plate. The thickness of the keratinized layer of the labial mucosa increased by 170% in KO mice and the muscle fiber diameter of the lingual muscle increased by 45%. Conclusion CKIP-1 genes have different effects on the growth and development of various soft and hard tissues in the maxillofacial region of mice.

Objective:To study the strategy of endovascular treatment for patients with the risks of internal carotid artery (ICA) rupture and bleeding during the treatment of nasopharyngeal carcinoma (NPC) based on American Society of Intervention and Therapeutic Neuroradiology/Society of Interventional Radiology (ASITN/SIR) grade of collateral circulation.Methods:A total of 56 patients (45 males and 11 females, aged from 28 to 76 years old) diagnosed with nasopharyngeal carcinoma and admitted to the Department of Neurosurgery, the Third Affiliated Hospital of Southern Medical University from July 2018 to January 2020 were retrospectively analyzed. There were 15 cases of ASITN/SIR grade 4, 24 cases of ASITN/SIR grade 3, 5 cases of ASITN/SIR grade 2, 5 cases of ASITN/SIR grade 1, and 7 cases of ASITN/SIR grade 0. The events of stroke and death were analyzed statistically.Results:ALL patients with ASITN/SIR grade 4 or 3 and some of patients with ASITN/SIR grades 2-0 passed balloon occlusion test and electrophysiological monitoring. ICA pseudoaneurysm was found in 35 patients, and one-stage ICA embolization was performed in 29 patients after evaluation. Among them, 8 cases of ASITN/SIR grade 4 and 10 cases of ASITN/SIR grade 3 with obvious posterior circulation compensation obtained successful one-stage ICA embolization without cerebral ischemia; cerebral ischemic events occurred in 5 (55.6%) of 9 patients with ASITN/SIR grade 3 and in 1(50.0%) of 2 patients with ASITN/SIR grade 2. The total incidence of ischemic events was 20.7% (6/29) and 1 case was disabled (1/29, 3.4%). Among patients with ASITN/SIR 3, there were statistically significant differences in stroke event rate between patients with obvious posterior circulation compensation and patients with slight or without posterior circulation compensation (0/10 vs. 5/9, χ 2=4.95, P=0.026). Follow-up time was 10.1±7.8 months, and 46 patients were survival (46/56, 82.1%) and 10 patients died (10/56, 17.9%) with a mean survival time of 2.6±1.4 months. Conclusions:For NPC patients with ICA invasion, ASITN/SIR based on DSA can simplify the assessment process of cerebral blood flow compensation. ICA can be embolized directly in patients with ASITN/SIR 4 or 3 with obvious posterior communicating compensation.

Objective: To analyze the clinical significance of contrast-enhanced ultrasound in the treatment of patients with carotid stenosis. Methods: The clinical data of 89 patients with carotid stenosis was retrospectively analyzed.The morphology and stenosis of carotid plaques were observed by contrast-enhanced ultrasound, and analyzing the relationship between the patient′s clinical symptoms and treatment options. Results: There were 66 males, 23 females, age ranging from 41 to 88 years.There were 147 plaques in 89 patients and 58 patients with bilateral lesions. The intensity of plaque ultrasound contrast was grade Ⅰ in 40 cases(27%), grade Ⅱ in 30(20%), grade Ⅲ in 31(21%), andgrade Ⅳ in 46(31%). The symptomatic group had higher CEUS strengths in grade Ⅲ(21.4%) and grade Ⅳ(37.9%). The difference was statistically significant between the two groups (P<0.05). Symptomatic group with high proportion of severe stenosis (44.7%) and occlusion (9.7%). It is narrower than that of asymptomatic group.The difference of stenosis was statistically significant between the two groups (P<0.05). Conclusion Contrast-enhanced ultrasound can dynamically and visually assess the distribution and density of carotid plaque morphology. It is useful for evaluating the treatment of patients with carotid stenosis.